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Breeding and Varietal Characteristics Analysis Of New Sugarcane Variety Mintang 11610 with Good Ratooning Early Maturity and High-Sugar Content
LI Ruimei, ZHANG Shuhe, LI Heping, HE Yansen
[Abstract](0) [FullText HTML](0) [PDF 565KB](0)
Abstract:
  Objective  To cultivate new sugarcane varieties with high yield, high sugar content, strong ratooning ability, and stress resistance, providing genetic support for the sustainable development of the sugarcane industry.  Method  A sexual hybridization breeding program was carried out using Yuenong 73-204 as the female parent and CP72-1210 as the male parent. the new variety Mintang 11610 was bred systematically through sexual hybridizatior and "Five-nursery System" breeding procedure. Then morphological characteristics and production characteristics were evaluated synthetically in the regional trial and production trial.   Results  The plants of Mintang 11610 were erect, medium to large stem; good seedling emergence, strong tillering ability, and a high stalk formation rate. It demonstrated strong lodging resistance and excellent ratooning ability.From 2015 to 2020, the average cane yield was 154.261 t·hm−2, which was increased by 15.11% compared with those of CK. The sucrose content was 16.30%, which was 1.43% higher than that of CK (absolute value).The yields of sugar was 25.145 t·hm−2, which increased by 26.18% compared with those of CK. During Perennial root cultivation, the yields of cane was 168.574 t·hm−2, which increased by 31.40% compared with those of CK, the sucrose content was 16.79%, which increased by 1.07% compared with that of CK (absolute value), the yields of sugar was 28.304 t· hm−2, which increased by 40.34%. Mintang 11610 was officially registered as a national non-major crop variety in December 2023, with the registration number GPD Sugarcane (2023) 350009.  Conclusion  Mintang 11610 is characterized by early maturity, high yield, high sugar content, good ratoonine abilities, The stability analysis showed that the stable high yield of this variety was better than that of CK, and the adaptability was wide.
Generating fresh sweet-waxy corn by using molecular marker-assisted selection technology
LU Xiaozhuan, ZHOU Shufen, LI Chenchen, CHEN Ziqiang, GUO Xinrui, Li Gang, TIAN Dagang
[Abstract](21) [FullText HTML](13) [PDF 1092KB](0)
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  Objective  To generate new variety of corn that has both sweet and waxy kernels on the same ear of sweet and waxy corn, explore new ways of rice breeding and improvement, and build the Fujian sweet-waxy corn brand.   Method  Using the main material of Jingkenuo 2000, a high-quality glutinous corn variety, we have developed the new germplasm GMC013 through pedigree breeding. This germplasm was hybridized with the sweet corn variety Xuetian 7401 and self-pollinated. By employing molecular marker-assisted selection and field selection methods, we have consolidated the sweet gene sh2 and the glutinous gene wx from the starch synthesis pathway in corn, creating a new high-quality sweet and glutinous fresh-eating corn germplasm.  Result  Molecular marker-assisted selection technology has successfully developed sweet variety GM102 and glutinous varieties GM104 and GM112. Field experiments indicate that these inbred lines meet the quality and appearance grain type standards for sweet and glutinous varieties in terms of growth duration, plant height, ear height, ear length, ear thickness, fresh weight of 100 grains, and seed setting rate.  Conclusion  The use of molecular-assisted selection techniques, combined with field selection, can precisely and efficiently create new superior maize germplasm, providing important genetic resources and reserves for maize variety breeding in Fujian.
Accumulation Characteristics of Total Phenolics, Flavonoids and Polysaccharides in Vegetable Jute Leaves of Different Growth Stages
HOU Wenhuan, LIAO Xiaofang, TANG Xingfu, ZHAO Yanhong
[Abstract](22) [FullText HTML](15) [PDF 572KB](0)
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  Objective  To study the accumulation characteristics of bioactive components such as total phenolics, flavonoids and polysaccharides in vegetable jute leaves at different growth stages, and to provide a reference for the rational harvesting, development and utilization of vegetable jute.  Method  Guimacai 1 and Guimacai 2 were used as experimental materials, and okra, Chinese yam and lettuce were used as control. The contents of total phenolics, flavonoids and polysaccharides in leaves of vegetable jutes were measured and analyzed at seedling stage, topping stage, flowering stage and capsule stage. The differences between vegetable jutes and control varieties were compared, and the correlation between growth stage and total phenolics, flavonoids, polysaccharides were analyzed.   Result  There were differences in the contents of total phenolics, flavonoids and polysaccharides in the same variety at different growth stages . With the extension of growth stage, the total phenolics content of the two vegetable jutes showed a trend of decreased first and then increased, and the total phenolics content in capsule stage was significantly higher than that in other growth stages, reaching 4.064 mg·g−1 and 3.852 mg·g−1, respectively. The total phenolics content of Guimacai 1 was higher than Guimacai 2 at seedling stage and capsule stage. With the extension of growth stage, the total flavonoids content of Guimacai 1 showed a trend of increased first and then decreased, and the highest content reached 2.755mg·g−1 at flowering stage. With the extension of growth stage, the total flavonoids content of Guimacai 2 showed an showed an increasing trend and the highest content in capsule stage was 4.755 mg·g−1, which was significantly higher than that in other growth stages.The total flavonoids content of Guimacai 2 was significantly higher than that of Guimacai 1 at the same growth stage.With the extension of growth stage, the polysaccharide content of Guimacai 1 showed showed a trend of fluctuated upward, the polysaccharide content of Guimacai 2 showed a trend of first decrease and then increase. The polysaccharide content of two vegetable jute varieties in the capsule stage was significantly higher than that in other growth stages, up to 3.175% and 1.240%, respectively. At the same growth stage, the polysaccharide content of Gammacai 2 was higher than that of Gammacai 1 only at the seedling stage. The contents of total phenolics and flavonoids of the two vegetable jute varieties were significantly higher than those of the control varieties at each growth stage, but the polysaccharide content was significantly lower than that of okra and Chinese yam, and the polysaccharide content was significantly higher than that of lettuce at capsule stage. Only the total flavonoid content of Guimacai2 was significantly positively correlated with the growth stage, while the others were not significantly correlated.   Conclusion  The vegetable jute leaves harvested at capsule stage were more conducive to the accumulation of total phenolics, total flavonoids and polysaccharides.
Differentiate Transcriptomes of Aerial and Creepy Agaricus bisporus Hyphae
DAI Jianqing, CHEN Wenzhi, ZENG Zhiheng, CAI Zhixin, CHEN Meiyuan
[Abstract](53) [FullText HTML](23) [PDF 1628KB](5)
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  Objective   Key genes associated with Agaricus bisporus of different mycelial morphology were analyzed. [Methods] As 2796, the main cultivated variety of A. bisporus, with aerial or creepy hyphae grown on same plate were collected for RNA transcriptome sequencing and comparison with reference.   Results   There were 965 differentially expressed genes (DEGs) between the aerial hyphae-A601 and the creepy hyphae-A602. According to the GO enrichment analysis, the hydrophobic DEGs were enriched in the fungal cell wall, such as AGABI2DRAFT_13655, AGABI2DRAFT_136569, and AGABI2DRAFT_193057 that belonged to the ABH3 protein gene family, with expressions significantly higher in A601 than in A602. Thus, they were postulated to be related to the production of aerial hyphae. In addition, the hydrophobic surface binding protein A (HsbA)-related gene, AGABI2DRAFT_194662, showed a FPKM expression of 0 in A601, but 10.24 in A602. Furthermore, 8 enriched DEGs related to lipase isoenzymes exhibited a FPKM expression significantly higher in A601 than in A602. The SNP/InDel analysis indicated that the SNPs in A601 was 110 601 as opposed to 111 188 in A602, the most frequent mutation in SNPs was C:G>T:A, and that 2 374 common InDels were identified.   Conclusion   The transcriptomes and possibly associated genes and SNP sites in A. bisporus with different mycelial morphology obtained in this study would be of value for the mushroom germplasm evaluation and breeding.
Characterization of vsiRNAs derived from Cymbidium Mosaic/Odontoglossum Ringspot Viruses in co-infected Phalaenopsis equestris
LAN Hanhong
[Abstract](18) [FullText HTML](14) [PDF 1177KB](0)
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  Objective  Interactions between Phalaenopsis equestris and cymbidium mosaic virus (CymMV) and odontoglossum ringspot virus (ORSV) were studied to aid the effort in developing effective preventive and control means against the diseases caused by the pathogens.   Method   The coat protein (CP) genes of CymMV and ORSV were amplified using RT-PCR. Under an electron microscope, viral morphology and size of CymMV and ORSV particles in P. equestris cells were examined. Abundance, length, base preference and origin of virus-derived vsiRNAs were analyzed applying the small RNA deep sequencing technology.   Result   The amplifications of CP genes of CymMV and ORSV were specifically obtain by RT-PCR. The electron microscopy revealed the lengths of the rod-like CymMV to be approximate 300 nm, while the linear ORSV, 500 nm. The small RNA deep sequencing yielded 7 563 892 CymMV-derived and 6 133 689 ORSV-derived vsiRNAs exhibiting the universality and specificity in abundance, length, base preference and sense strand distribution.   Conclusion  Co-infections of CymMV and ORSV on P. equestris were clearly demonstrated in this study. The vsiRNAs of CymMV and ORSV displayed characteristic patterns in abundance, length, base preferences and sense strand distribution.
Disease and Insect Resistance Genes in Premium Sterile Line Funong A
LIAN Ling, TU Shihang, ZHANG Junian, DONG Meng, HE Wei, ZHENG Yanmei, XIE Zhenxing, SHI Longqing, JIANG Zhaowei, WU Chunzhu
[Abstract](35) [FullText HTML](20) [PDF 1739KB](6)
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  Objective  Disease and insect resistance genes in Funong A, a three-line indica cytoplasmic male sterile line, were analyzed for breeding applications.  Method   Genomic DNA of Funong A, Fudao B, and Huahangsimiao rice germplasms was extracted by CTAB method followed by detection of blast resistance genes with molecular markers. Resistance genes related to bacterial blight, viral diseases, and brown planthopper infestation were analyzed by high density chip detection method. Blast resistance genes of the lines were obtained by PCR and sequenced for comparison. Specimens were collected at 0, 24, 48, 72, and 96h after a blast fungus inoculation for extraction of total RNA using Trizol method. Gene expression was determined by SYBR Green I qRT-PCR.   Result  Funong A, Fudao B, and Huahangsimiao were found to contain the blast resistance genes Pib, Pia, Pid3, Pid2, bacterial blight resistance gene Xa21, and yellow mottle virus resistance gene Rymv1. Furthermore, Funong A and Huahangsimiao had the blast resistance gene Pi5 and the stripe virus resistance gene STV11, while Fudao B consisted of the blast resistance genes Pita and Pi37. However, no brown planthopper resistance genes were detected in them. The fragments of Pi5, Pia, Pib, Pid3, and Pid2 were 5,672 bp, 2,597 bp, 5,532 bp, 2,865 bp, and 3,847 bp in length, respectively. The sequences of Pi5, Pid3, and Pid2 in Funong A were similar to those in Huahangsimiao, but 2 bases differed from Pid3 and 5 from Pid2 of Fudao B. In Funong A, the expressions of rice blast resistance genes Pi5 and Pib were significantly induced by the blast fungus with a peak occurred 72h after inoculation. The expressions of Pia and Pid3 increased gradually with time after inoculation to peak in 96h. The expression of Pid2 rose initially to a maximum in 24h and then declined.  Conclusion  The three-line indica cytoplasmic male sterile line Funong A carried the blast resistance genes Pi5, Pib, Pia, Pid3, and Pid2, which were significantly induced by the blast fungus. The line also contained the bacterial blight resistance gene Xa21, yellow mottle virus resistance gene Rymv1, and stripe virus resistance gene STV11 but none of the insect resistance genes.
Differentiation in Post-harvest Lipid Metabolism of Oil Palm Fruits
LI Xinyu, LIU Xiaoyu, LI Qihong, ZHOU Lixia, LI Rui, FU Dengqiang, CAO Hongxing
[Abstract](23) [FullText HTML](20) [PDF 2407KB](0)
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  Objective   Mechanism and accumulation of lipid synthesis in thin-shelled oil palm fruits was investigated for breeding of variety resistant to fat rancidity.   Method   Fruits of thin-shelled oil palm freshly harvested 185d after pollination (T1), 24h post-harvest (T2), and 36h post-harvest (T3) were collected for LC-MS/MS and RNA-seq determination and analysis on lipid metabolites and differentially expressed genes in mesocarp of oil palm fruits as the lipid oxidation taking place. [Result] In the fruit development, 5 lipid classes, 23 lipid subclasses, and 520 monomer molecules in mesocarp of the oil palm fruits were identified. It is well known that the hydrolysis of phosphatidylcholine (PC) is a lipid oxidation and the hydrolyzed glycerophosphate choline (GPC) affects PC, lipophosphatase (LPP) promotes synthesis of phosphates and glycerophospholipids, and the expression of chlorophyll relates to chlorophyll content. This study found that the aldehyde dehydrogenase (ALDH7A1 and ALDH2), monoacylglycerol lipase (MGL), phospholipase A1 (PLA1), and glycerophosphodiester phosphodiesterase (GDPD1) significantly negatively correlated with glycerophospholipids, such as diacylglycerol trimethyl homoserine (DGTS), phosphatidic acid (PA), phosphatidylinositol (PI), phosphatidylcholine (PC), phosphatidylglycerol (PG), phosphatidylethanolamine (PE), but significantly positively correlated with palmitic acid, while GDPD1, LPP, and digalactosylglycerol synthase (DGD1) significantly positively correlated with the glycerophospholipids, such as DGTS, PA, PI, PC, PG, PE, but negatively correlated with palmitic acid, whereas MGL monoglyceride (MG) extremely significantly positively correlated with linoleic acid (LA) but significantly negatively correlated with ceramide (Cer), and DGD1 and LPP significantly negatively correlated with MG and LA, but significantly positively correlated with Cer.   Conclusion   It appeared that ALDH7A1, ALDH2, PLA1, and MGL inhibited the glycerophospholipids synthesis but promoted the synthesis of fatty acids such as palmitic acid, while DGD1, LPP, and GDP1 enhanced the synthesis of glycerophospholipids but retarded that of palmitic and other fatty acids.
Vector Construction and Immunogenicity of S and N Gene DNA Vaccine for TGEV
CHENG Weiwei, RONG Weizhong, YANG Ming, LI Yuanxin, ZHAO Zihui, CHEN Boxiang, WANG Jia, ZHOU Yao
[Abstract](42) [FullText HTML](27) [PDF 1580KB](0)
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  Objective   DNA vaccine vector of S and N genes of porcine transmissible gastroenteritis virus (TGEV) was constructed with the vaccine immunogenicity detemined to pave the way for studying, preventing, and controling TGE.  Method   A and D sites on S and N from a TGEV were amplified. The N gene alone as well as the A and D sites fusion were cloned into the vaccine vector pCDNA3.1-His-C. Bioinformatics software was used to predict and analyze the secondary structure, tertiary configuration, subcellular localization, and dominant B cell epitope of S (A-D) and N proteins. The recombinant vectors were transfected into PK-15 cells, and expression distribution of N and the A and D sites fusion detected by indirect immunofluorescence and confocal detection. Mice were immunized with the single or combined recombinant vaccine vector to detect the IgG antibody using indirect ELISA.  Result   The A and D sites of the S were 498bp and 606bp, respectively, and the N, 1,149bp in length. The nucleic acid vaccine expression vectors p-S (A-D)-His and p-N-His for the A and D sites (fusion) and N were constructed. Bioinformatics software predicted that, when TGEV infected the host cells, N protein was mainly located in the nucleus and mitochondria and S (A-D) largely in the cytoplasm and mitochondria, while S (A-D) had 7 and N, 8 dominant B cell epitopes. All p-S (A-D)-His and p-N-His were successfully expressed in PK-15 cells distributed in the nucleus and cytoplasm. The immunized mice showed an effect of immunity in the order of p-N-His>p-S (A-D)-His + p-N-His>p-S (A-D)-His.   Conclusion  The DNA vaccine vectors of S and N of TGEV were successfully constructed. Strong specific antibodies were generated in lab mice after the immunization.
Optimization of Astaxanthin-producing Fermentation by Phaffia rhodozyma using Okara as Nitrogen Source
GUAN Xuefang, CHEN Shuying, LAI Gongti, WANG Qi, LAI Chengchun, HUANG Juqing, ZHENG Qi, LIN Bin
[Abstract](64) [FullText HTML](23) [PDF 997KB](5)
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  Objective   An astaxanthin-producing fermentation by Phaffia rhodozyma using okara for nitrogen was optimize.   Methods   On the conventional fermentation by P. rhodozyma to make astaxanthin, okara was used to replace the commonly applied peptone and yeast extract as the organic nitrogen source for cost reduction. Effects of carbon sources, precursor substances, other nitrogen supply, vitamins, and inorganic salts on yield of astaxanthin were analyzed with the amounts of (NH4)2SO4, vitamin E, glucose, and sucrose optimized by response surface methodology.   Results   When okara was used as a raw ingredient for the fermentation, glucose became the optimal carbon source. The yield of astaxanthin by the P. rhodozyma fermentation could be significantly increased by applying both glucose and sucrose, potassium salts such as KCl, KNO3, and K2HPO4 as well as (NH4)2SO4, VB2, VE, and zeaxanthin. Hence, the medium was optimized by response surface method on the 4 key ingredients of glucose, sucrose, K2SO4, and VE to arrive at a formulation consisting of 10% okara, 0.22% K2SO4, 0.6% VE, 1.08% glucose, and 1.50% sucrose to reach a yield of astaxanthin at 32.46 mg·L−1, which was 2.23 folds higher than what obtained by using the YM medium.   Conclusion   Okara could amply be used to replace peptone and yeast extract as the nitrogen source for the astaxanthin production by P. rhodozyma fermentation.
A Constant Temperature Fluorescent RPA Assay for Detecting Novel Muscovy Duck Parvovirus
CHEN Wei, LIANG Qizhang, ZHANG Jiaxue, JIAO Wenlong, LIN Yongqiang, LIU Rongchang, FU Qiuling, FU Guanghua, ZHU Ting, HUANG Yu
[Abstract](44) [FullText HTML](17) [PDF 981KB](2)
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  Objective  A constant temperature fluorescent RPA assay for detecting novel Muscovy duck parvovirus (N-MDPV) was developed.   Method   EXO fluorescent probes were specifically bound to the targeted conserved VP3 fragment of N-MDPV. RPA amplification primers were designed to amplify the segment by using recombinant enzyme polymerase. An assay for detecting N-MDPV was established with reaction time and temperature optimized and specificity, sensitivity, and accuracy tested on the collected nucleic acid of disease material in comparison with traditional PCR and virus isolation identification methods.   Result  The optimized assay reaction temperature and time were 39 ℃ for 30m to achieve a lowest sensitivity for nucleic acid detection at 10 fg·μl−1. The nucleic acid of N-MDPV was specifically amplified without any cross reaction with those of duck adenovirus type 3, avian adenovirus type 4, duck circovirus, duck plague virus, duck viral hepatitis virus, duck Tambousu virus, and novel duck reovirus. Along with the conventional PCR and the virus isolation and identification methods, the newly developed assay detected the nucleic acid on 38 duck tissue specimens with a positive rate of 36.8% (14/38) in comparison to those at 36.8% (14/38) for the PCR and 31.6% (12/38) for the isolation and identification method. In addition, 100% coincidence rates of the assay and the two other methods on positive samples as well as of the assay and the PCR on the positive clinical samples were secured.   Conclusion   The new RPA method to rapidly and visually detect N-MDPV demonstrated to be highly specific, sensitive, and accurate. It was deemed appropriate for clinical applications.
Full-length Transcriptome of She Herbal Medicine Plant Clematis florida
HE Shulan, KE Lanlan, CHEN Guanshui
[Abstract](21) [FullText HTML](16) [PDF 1642KB](0)
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  Objective   Transcription of herbal plant Clematis florida Thunb. var. plena D. Don was used to identify the key genes involved in the metabolic pathways of the active compounds in the famed “Shiershichen” of She medicine.  Method   PacBio single-molecule real-time (SMRT) was employed to sequence the full-length transcriptome of the roots of the herbal plant that contains the active compounds of the She medicine. Functional annotation, gene structure, and mining of the terpenoid biosynthetic pathway were conducted using bioinformatics tools to secure the transcript data.  Result  From the 62.21 G polymerase read bases generated, 20,540 non-redundant high-quality transcript sequences were identified after data treatment. At least one of the 7 major databases including NR, NT, Pfam, COG/KOG, SwissProt, GO, and KEGG applied annotated the gene functions of 19,909 transcripts, and 8,888 were in NR, NT, COG/KOG, KEGG, and GO. The GO annotation showed 14,911 transcripts enriched in 53 terms that included biological processes, cellular components, and molecular functions. The KEGG database annotated 19,701 transcripts and classified them into 6 major pathways and 44 sub-pathways with the largest number of transcripts enriched in metabolic pathways. The COG/KOG annotation identified 13,204 transcripts with the general function prediction being the most predominant. There were 978 transcription factors, 224 lncRNAs, and 7,167 SSRs predicted, and 48 transcripts with 16 key candidate genes involved in the terpenoid backbone biosynthesis identified.  Conclusion  The full-length transcriptome sequencing on the roots of C. florida was successfully obtained with the comprehensive gene function information. It provided a basis for further studies on the regulatory network, biological characteristics, related metabolic pathways, signaling pathways, and molecular mechanisms associated with the well-known She herbal medicine.
Risk Assessment on Resistance to Pyraclostrobin of Colletotrichum gloeosporioides
RUAN Hongchun, HUANG Yanqin, CHEN Qiaohong, CHEN Wenle, LAN Chengzhong
[Abstract](19) [FullText HTML](12) [PDF 743KB](0)
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  Objective   Drug sensitivity to and cross-resistance of pyraclostrobin with other fungicides of Colletotrichum gloeosporioides that caused anthracnose in soybeans in Fujian Province were investigated.   Methods  Sensitivity to pyraclostrobin of 112 strains of C. gloeosporioides isolated from 5 regions in the province in 2023 was determined using the mycelial production rate method. Resistant mutants of the pathogen were obtained by fungicide domestication to determine their genetic stability, fitness, and possible cross-resistance to 4 fungicides.   Results   The EC50 of pyraclostrobin for the 112 C. gloeosporioides strains ranged 0.0682 µg·mL−1 to 1.0309 µg·mL−1 with a coefficient of variation of 15.12 and averaged 0.203 6±0.121 5 µg·mL−1 as the sensitivity baseline. The distribution of the sensitivity frequency was in a continuous unimodal function consistent with the continuous skewed normal distribution. Induced from two wild C. gloeosporioides, 4 moderate resistant strains and one low resistant strain were secured at a frequency of 8.3×10−4. The mutants stably inherited the drug resistance traits but differed from their parents in temperature sensitivity and were slightly lower on mycelial growth rate, sporulation ability, and pathogenicity. In the field, they might sacrifice other natural competitive advantages as a fitness cost in exchange for the drug resistance. Pyraclostrobin showed no cross-resistance with carbendazim, difenoconazole, and prochloraz but did with the same type of fungicide, picoxystrobin.  Conclusion  Although the C. gloeosporioides isolated in Fujian were relatively high in sensitivity to pyraclostrobin, they represented merely a moderate drug resistance risk. Nonetheless, in practice, it was plausible to blend pyraclostrobin or apply alternatively with fungicides such as carbendazim, difenoconazole, and prochloraz to delay such potential drawback.
Genetic Diversity and Principal Components of Tea Germplasms from Shiting Plantations in Nan'an
OU Xiaoxi, LIN Hongzheng, HE Jihang, LI Qiuming, FU Ruidian, CHEN Yucai, ZHENG Yucheng, SUN Yun
[Abstract](28) [FullText HTML](13) [PDF 1900KB](0)
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  Objective  Genetic diversity of the tea germplasms preserved at Shiting plantations in Nan’an was studied for promoting the utilization of the local specialty.  Methods  SNP molecular marker technology was applied to analyze the genetic diversity and relationships of 17 tea germplasms from the Shiting plantations and 6 representative varieties from Fujian. HPLC and UPLC-MS/MS were employed to determine the contents of catechins and free amino acids in 10 selected samples for a comparison among the tea varieties.  Results  Forty-four SNP loci suitable for identifying the tea plants were selected. An SNP fingerprint profile of the germplasms was constructed. The UPGMA evolutionary tree divided the 23 germplasms into 4 subgroups, which showed a close relation with Rougui and Qidan teas from northern Fujian. Significant differences were found on the contents of major amino acids and catechins among the varieties (P<0.05) with total free amino acids from 8.46 mg·g−1 to 32.66 mg·g−1 and catechins ranging from 107.56 mg·g−1 to 177.60 mg·g−1. The germplasms coded ST2, ST3, ST6, and ST17 had high contents of EGCG or EGCG3"Me, while ST3 the most on amino acids and the least on catechin bitterness index.   Conclusion   Shiting plantations in Nan’an had a rich collection of tea germplasms that were closely related to those in northern Fujian. Among them, ST2, ST3, ST6, and ST17 teas had high contents of either EGCG or EGCG3"Me. ST3, specifically, appeared to be suitable for making green tea as well as in breeding programs.
Establishment and application of multiplex RT-PCR methods for akabane virus, bovine herpesvirus 1 and bovine viral diarrhea virus
ZHANG Fanfan, WEI Jiazhen, LI Jiemao, TAN Meifang, HU Lizhen, ZENG Yanbing, GONG Xiaoqi, YANG Qun, XU Jun
[Abstract](23) [FullText HTML](19) [PDF 743KB](0)
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  Objective  To establish a multiplex RT-PCR method for the rapid and simultaneous detection of akabane virus (AKAV), Bovine herpesvirus 1 (BoHV-1) and bovine viral diarrhea virus (BVDV), so as to lay a foundation for the differential diagnosis, prevention and control of bovine reproductive disorder epidemics.   Method  Three pairs of primers were designed for the conservative regions of AKAV S gene, BoHV-1 gH gene and BVDV 3'UTR gene, and a multiplex RT-PCR assay for AKAV, BoHV-1 and BVDV was established through the optimal of reaction condition parameters.   Result  The specificity results showed that the method was positive only for AKAV, BoHV-1 and BVDV, and negative for the nucleic acids of bovine parvovirus (BPV), bovine respiratory syncytial virus (BRSV), bovine parainfluenza virus 3 (BPIV3), foot-and-mouth disease virus (FMDV), and bovine ephemeral fever virus (BEFV). The sensitivity results showed that the lower threshold of this method for AKAV, BoHV-1 and BVDV recombinant plasmids were all 1.0×103 copies/μL. The results of the reproducibility test showed good intra- and inter-batch reproducibility. The established method was used to determine 143 whole blood/tissue samples of cattle with reproductive disorders and compared with the existing local standards to verify the efficacy of this test for practical clinical application. The results showed that the positivity rates of AKAV, BoHV-1 and BVDV were 2.80% (4/143), 21.68% (31/143) and 38.46% (55/143), respectively, with the presence of several mixed infections; and the conformity rates of this method with the local standards for AKAV, BoHV-1 and BVDV were 99.3%, 98.6% and 97.2%, respectively. The Kappa coefficients of the two were 0.885, 0.960 and 0.942 respectively, indicating that the method established in this study was in good agreement with the local standards.   Conclusion  In this study, we pioneered a multiplex RT-PCR assay with high specificity, sensitivity and low cost for the differential detection of AKAV, BoHV-1 and BVDV.
Establishment and preliminary application of fluorescent RT-RAA diagnostic method for porcine epidemic diarrhea virus
HOU Enhui, CHEN Qiuyong, WU Xuemin, QIU Jingli, WU Renjie, ZHOU Lunjiang, LIU Yutao, MA Yufang, WANG Longbai
[Abstract](21) [FullText HTML](15) [PDF 1973KB](1)
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  Objective  To establish a quick, simple, sensitive detection of porcine epidemic diarrhea virus (PEDV) the RT- RAA detection method, to improve the efficiency of porcine epidemic diarrhea virus clinical detection.   Method  Primers and probes were designed for the conserved region of PEDVS gene fragment, and a standard plasmid PEDV-S was constructed. Through specificity, sensitivity, repeatability and condition optimization, a recombinant enzyme-mediated chain replacement nucleic acid amplification fluorescence assay (RT-RAA) for detection of PEDV was established.   Result  Under the condition of constant temperature at 42 ℃ for 20 min,The established assay is effective for the detection of porcine transmissible gastroenteritis virus (TGEV), classical swine fever virus (CSFV), porcine pseudorabies virus (PRV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine rotavirus (PoRV) and other porcine viruses were negative, porcine epidemic diarrhea virus (PEDV) was positive.The minimum detection limit was 4.43× 102 copies·μL−1 standard plasmid.The reproducibility results showed that there was little difference between the standard plasmids with the same concentration.The positive rate of 40 swine virus samples was 7.5% (3/40) by RT-RAA method, which was the same as that of real-time fluorescence quantitative PCR.   Conclusion  The RT-RAA detection method for PEDV established in this study is suitable for the rapid diagnosis of PEDV due to its rapid and simple detection, short time-consuming, high specificity, strong sensitivity and good reproducibility.
Metabolomics analysis of mulberry fruits at different developmental stages
ZHONG Qiuzhen, LIN Qihua, ZHANG Zehuang
[Abstract](23) [FullText HTML](14) [PDF 1816KB](2)
Abstract:
  Objectives  To study the changes in metabolic components at different development stages of mulberry fruits.   Methods  a widely targeted metabolomics analysis was conducted on samples from the green fruit stage, color transformation stage, and mature stage of Morus alba cv. yueshen dashi. And the KEGG metabolic pathway enrichment was then analysed based on differentially accumulated metabolites (DAMs).  Results  A total of 1146 metabolic components were identified from three developmental stages of mulberry fruit. Using P<0.05 and VIP>1.0 as the criteria, 483 metabolites were screened as DAMs, including 51 DAMs with significant differences in accumulation levels throughout the whole comparison groups. Two flavonoids and one anthocyanin were significantly increased during the development of mulberries. DAMs based KEGG metabolic pathway analysis found that α-linolenic acid metabolism and linoleic acid metabolism were significantly enriched during the green fruit development to color transition period, while ascorbate and aldarate metabolism were significantly enriched during the color transition period to maturity period, while linoleic acid metabolism and cutin, suberine and wax biosynthesis were significantly enriched throughout the fruit development process. Further analysis revealed that although there were six upaccumulated and one downaccumulated linoleic acid components in the development process of mulberry fruit, the overall accumulation level of linoleic acid showed a downward trend.  Conclusions  This study found that 51 DAMs might involve in the entire development process of mulberry fruits, the anthocyanins and flavonoids were significant increased, and 7 key metabolic components in the linoleic acid metabolism pathway may affect the formation of its quality and flavor. These results contributed to a better understanding of the dynamic changes in nutritional components during the ripening process of mulberries, and laid the foundation for revealing the mechanism of mulberry fruit quality and flavor formation and screening high-quality mulberry germplasm. Additionally, this research provided scientific basis for determining the picking period of mulberries.
Proteomics analysis on response of Duyun Maojian native tea plant to the invasion of Empoasca vitis Gothe
WANG Fen, YU Pei, ZHAI Yuan, CHUANGANG Li, LUO Siyu, PEI Huimin
[Abstract](33) [FullText HTML](25) [PDF 1151KB](1)
Abstract:
  Objective  In order to study the proteins related to the response of Duyun Maojian native tea plant infected by Empoasca vitis Gothe.  Method  The leaves of Duyun Maojian native tea plant were infected by E. vitis Göthe for 0 h, 12 h, 24 h, 36 h and 48 h, were used as research materials. Tandem mass tag (TMT) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) technique was used to qualitatively and quantitatively analyze the proteins of tea leaves infected with E. vitis Göthe.   Results  We identified a total of 2,893 proteins. There were 0, 1, 848, and 849 differential expression proteins (DEPs) between 0 h and 12 h, 0 h and 24 h, 0 h and 36 h, 0 h and 48 h, respectively. Furthermore, 2,622 proteins were annotated, of which 2,360 proteins were annotated to the GO database and 1,232 proteins were annotated to KEGG database. GO and KEGG analyses showed that tea plants may be in resistance to E. vitis Göthe infestation through CMK, FPPS, GPPS, GGPPS, peroxidase, pectinesterase, and allene-oxide cyclase, sesquiterpenoid, triterpeoid, monoterpenoid, diterpenoid, sesquiterpenoid and triterpenoid biosynthesis, terpenoid backbone biosynthesis, and monoterpenoid biosynthesis.   Conclusion  Four DEPs (CMK, FPPS, GPPS, and GGPPS) and terpenoids may play important roles in the response and defense against E. vitis Göthe. The research results may provide theoretical basis for revealing the molecular mechanism of tea plant response to the harm of E. vitis Göthe.
Identification and Evaluation of Stem Rot Antagonistic and Growth promotion Effects of Trichoderma on Anoectochilus roxburghii
YE Wei, YAN Peipei, WAN Peiyu, LI Zunwen, GUO Yin, ZHANG Jianliang, LIN Minshui, JIANG Jinlan
[Abstract](28) [FullText HTML](16) [PDF 2580KB](1)
Abstract:
  Objective  Isolation of stem rot antagonistic Trichoderma from Anoectochilus roxburghii, providing theoretical basis for the development of biocontrol fungi. in A. roxburghii.  Method  Using wild cultivated A.roxburghiias as materials, Trichoderma strains were isolated using tissue isolation method. Morphological characteristics and homology analysis with ITS and rpb2 sequences were conducted for strains classification. Plate confrontation method were used for evaluating different Trichoderma strains resistance to stem rot ability, and the growth promoting effects of different Trichoderma strains were also processed.[Resulsts]3 Trichodermas trains as A21B-1, A21B-2 and A21E were isolated from the A. roxburghii via tissue isolation method. Combined with morphological characterization and ITS and rpb2 sequences homology identification, 3 Trichoderma strains were identified as T. rugulosum, T. koningiopsis and T. longifialidicum, respectively. Confrontation cultured showed that the 3 Trichoderma strains showed strong inhibitory effects on stem rot pathogen Fusarium oxysporum f. sp. opponiarum ASP01, and their inhibition rates reached 75.29%, 73.55% and 60.02%, respectively. The indoor control results showed that A21B-1 strain had a strong inhibitory effect on stem rot, after 15 d inoculation, the disease inhibition rate reached 91.9%, which could be used as a candidate strain for stem rot biological control fungi. The growth promotion experiments showed that Trichoderma strains significantly increased individual plant weight, height, stem diameter, leaf area, and SPAD value in A. roxburghii after 6 months grown. Among the 3 Trichoderma strains, plants innoculated with A21B-2 and A21E showed significant growth promotion effects with individual plant weight increased by 58.6% and 58.9%, leaf area increased by 66.8% and 59.7% compared to the controls, respectively. They could be used as candidate strains for growth promoting in A. roxburghii. At the same time, the application of Trichoderma strains effectively increased the content of polysaccharides and kinsenoside in A. roxburghii, A21B-2 strain showed the best effects, which content of polysaccharides and kinsenoside increased by 89.6% and 11.8% compared to the controls, could be used a candidate strain for promoting the accumulation of medicinal components in A. roxburghii.  Conclusion  3 different strains of Trichoderma have significant effects on inhibiting stem rot disease, promoting growth, and increasing polysaccharide content in A. roxburghii.
OsPLGG1 enhanced function of photorespiratory bypass
CUI Lili, CAI Qiuhua, QIU Jieyu, GAO Rongrong, ZHAO Yongchao, WANG Yingheng, PENG Xinxiang, ZHU Guohui, ZHANG Jianfu
[Abstract](30) [FullText HTML](23) [PDF 2772KB](3)
Abstract:
  Objective  The deletion of either OsPLGG1a or OsPLGG1b in the GOC rice engineering optimizes the photorespiration pathway, thereby enhancing rice photosynthesis efficiency and optimizing photorespiration metabolic engineering.  Method  The knockout vectors osplgg1a-Cas9 or osplgg1b-Cas9 were constructed and transformed into GOC rice, respectively, to obtain GOC transgenic homozygous lines with deletion of OsPLGG1a or OsPLGG1b, and the photosynthesis rate was measured.   Result  Transgenic homozygous lines with the GOC rice of OsPLGG1a or OsPLGG1b knocked out were obtained. The phenotypes of osplgg1a-GOC plants were similar to osplgg1a, both of which showed stunted growth. Compared with GOC rice, the net photosynthetic rate of osplgg1b-GOC plants increased, indicating that OsPLGG1b mutation was conducive to the retention of chloroplast glycolate, increased the metabolic flux through the GOC bypass, reduced the photorespiration metabolism of plants, and further increased the CO2 concentration in chloroplasts.   Conclusion  Compared with GOC rice, the net photosynthetic rate and stomatal conductance of osplgg1b-GOC plants were increased, suggesting that the elimination of OsPLGG1b could be used to optimize the photorespiratory metabolic pathway.
Effects of the density of the flea beetle Agasicles hygrophila on the clonal integration of alligator weed Alternanthera philoxeroides
LI Yuehua, ZHOU Yuhang, WU Haohai, ZHENG Wei, LI Xiaoqiong
[Abstract](24) [FullText HTML](20) [PDF 844KB](0)
Abstract:
  Objective  Many invasive plants have clonal growth habits, and clonal integration is considered as an effective way for clonal invaders to alleviate the feeding pressure of natural enemies. However, the effects of the density of natural enemies on the clonal integration ability of invasive clonal plants are largely unclear.  Methods  In this study, the invasive alligator weed Alternanthera philoxeroides was used as the study object. A common garden experiment was carried out to compare the effects of clonal integration on the growth traits, root growth and biomass allocation of the apical and basal ramets as well as the entire clonal fragment of alligator weed under different feeding densities of the flea beetle Agasicles hygrophila.  Results  The number of leaves, aboveground biomass, belowground biomass, total biomass and coarse roots of apical ramets as well as belowground biomass of the entire clonal fragment of alligator weed were significantly lower under herbivory by flea beetles compared to without herbivory. The number of leaves, coarse roots, total roots, aboveground biomass, belowground biomass and total biomass of the apical ramets, the diameter of the basal ramets as well as the ground diameter, aboveground biomass, belowground biomass and total biomass of the entire clonal fragment of alligator weed were significantly higher with clonal integration compared to without clonal integration. Under herbivory by one flea beetle, the number of coarse roots of the apical ramets, the ground diameter of the basal ramets and the ground diameter, the number of coarse roots and aboveground biomass of the entire clonal fragment of alligator weed were significantly higher with clonal integration compared to without clonal integration. However, when under herbivory by two flea beetles, although there were no significant differences in the ground diameter, number of coarse roots and aboveground biomass of the apical and basal ramets, the number of leaves, stem length and ground diameter of the entire clonal fragment were significantly higher, with clonal integration compared to without clonal integration.  Conclusion  The flea beetle density significantly affected the clonal integration ability of alligator weed: this plant can benefit significantly from clone integration without herbivory or under relatively lower flea beetle density (one beetle per plants); however, relatively higher density of flea beetles (two beetles per plant) can conversely reduce the clonal integration ability of alligator weed, thus achieving effective biological control of the flea beetle on alligator weed.
Genetic Diversity and Relationship Analysis of Gerbera jamesonii Bolus Germplasm Resources Based on EST-SSR Markers
CAO Yiyang, XIA Chaoshui, CHEN Weiting, GAN Weixin, LIN Fazhuang, XU Kezheng, LIN Huifeng
[Abstract](24) [FullText HTML](11) [PDF 1815KB](1)
Abstract:
  Objective  To reveal the genetic diversity and relationship of 170 germplasm resources of Gerbera jamesonii Bolus in different populations and types, and provide reference for introduction, protection and utilization of gerbera germplasm resources.  Method  1-3 pairs of EST-SSR primers with high polymorphism and clear bands were selected for each chromosome pair, and 170 gerbera germplasm DNA from 6 different populations were amplified by PCR. SSR polymorphism, genetic differentiation among different populations, genetic uniformity and UPGMA cluster analysis were used to evaluate the genetic diversity and phylogenetic relationships among different individuals and populations of gerbera.   Results  A total of 168 alleles (Na) were detected in 39 selected EST-SSR primers, with an average number of 4.308, the average Shannon information index (I) was 1.098, and the variation of polymorphism information content (PIC) was 0.431-0.920, with an average of 0.760, which was higher than 0.5. The total allele number, total genotype number, average allele number, average genotype number, and average heterozygosity were higher in population of China Yunnan and mixed population, the genetic diversity was relatively rich. The genetic distance of the six populations ranged from 0.016 to 0.158, with an average of 0.069, and the genetic consistency ranged from 0.854 to 0.984, with an average of 0.935. The genetic distance between China Yunnan population and mixed population was the smallest, and the genetic distance between German population and Japanese population was the largest. The results of population clustering analysis showed that the German population, China Yunnan population and mixed population were clustered into one group, and the relationship was relatively close. The results of individual clustering showed that 170 germplasm resources were divided into 6 groups (I, II, III, IV, V, and VI) at the genetic similarity coefficient of 0.550. At the genetic similarity coefficient of 0.558, group V could be divided into 4 subgroups, and at the genetic similarity coefficient of 0.570, group VI could be divided into 4 subgroups. The distribution of gerbera with single population origin, including types of potting, pasta, bubble, was relatively simple, while the distribution of other gerbera germplasm was relatively dispersed.   Conclusion  The EST-SSR markers were highly polymorphic and could be used to analyze the genetic diversity and relatives of gerbera germplasm resources. The genetic diversity of gerbera germplasm resources is rich, and the genetic diversity of different populations is quite different. The results of this study could provide important references for the introduction, protection and breeding utilization of gerbera germplasm resources.
Transcriptome and Metabolome Analysis of Tea Leaves in Response to Tea Blister Blight
YAO Yuxian, ZHANG Mingze, SHU Ye, LUO Fudan, QUAN Changbin, LIU Rong, XIANG Zhipan, MEI Xin
[Abstract](25) [FullText HTML](13) [PDF 3302KB](0)
Abstract:
  Objective  To explore the resistance molecular mechanism of tea plants to the pathogen of tea blister blight, and to explore the resistance-related genes, so as to provide a basis for resistance breeding of tea plants.  Methods  Differentially expressed genes (DEGs) and differentially accumulated metabolites (DAMs) were compared between healthy leaves (CK) and leaves infected with blister blight disease (TB) by transcriptome sequencing and metabolome analysis.  Results  Transcriptome data showed that there were 1009 DEGs between CK and TB, and GO enrichment analysis indicated that the DEGs were involved in cell wall metabolism and the regulation of chitinase activity, oxidoreductase activity, and xyloglucan:xyloglucanosyltransferase activity. KEGG metabolic pathway analysis showed that the DEGs were significantly enriched in pathways of "Flavonoid biosynthesis", "Phenylpropanoid biosynthesis", "Amino sugar and nucleotide sugar metabolism", "Glycerolipid metabolism", and "Stilbenoid, diarylheptanoid and gingerol biosynthesis". 47 transcription factors in DEGs belonging to 21 transcription factor families, mainly including bHLH, SBP, AP2/ERF-AP2 and MYB, etc., which may be important regulatory genes in the process of tea plant to blister blight disease. A total of 353 DAMs were identified using widely targeted metabolomics, and the DAMs were mainly enriched in the pathways of "Flavonoid biosynthesis", "Lysine biosynthesis", and "Alanine, aspartate and glutamate metabolism". Integrative analysis of transcriptome and metabolome revealed that the pathways with significant co-enrichment were "Flavonoid biosynthesis", "Phenylpropanoid biosynthesis", "Stilbenoid, diarylheptanoid and gingerol biosynthesis". A total of 20 DEGs and 15 DAMs related to the phenylpropanoids and flavonoids biosynthesis pathways were screened out. Among them, DEGs such as CSS00117414CL)、CSS0002940DFR)、CSS0015968DFR)and CSS0010687ANS)were up-regulated in susceptible leaves. DAMs such as phloretin, phlorizin, 4-Hydroxystyrene, p-Coumaroyl quinic acid, dihydromyricetin, epigallocatechin and peonidin 3-O-glucoside were accumulated in susceptible leaves.   Conclusion  DEGs in the pathways of "Phenylpropanoid biosynthesis" and "Flavonoid biosynthesis" play important roles in the response of tea plant to tea blister blight disease infestation, and DAMs such as phloretin, phlorizin, and epigallocatechin may be important secondary metabolites for tea plant resistance to blister blight disease.
Effect of arbuscular mycorrhizal fungi on the growth and secondary metabolism of Taxus chinensis (seedling stage )
NING Xiangjun, LI Man, CHEN Xiaole, FENG Yu, CHEN Yuxin, TU Hao, HE Jiayue, WU Fangjing, LIU Hongling
[Abstract](22) [FullText HTML](12) [PDF 963KB](0)
Abstract:
  Objective  To investigate the impacts of AM fungi on the growth and secondary metabolism of Taxus chinensis seedlings, to discover the symbiotic relationship between Taxus chinensis and AM fungi, and to offer a scientific foundation for Taxus chinensis planting and use.  Method  Taxus chinensis seedlings were used as materials, and the roots were inoculated with the arbuscular mycorrhizal fungi (AMF) Rhizophagus intraradices and Funneliformis mosseae during the potting test, as well as a mixture of the two strains, and the inoculation was timed to coincide with planting in this experiment. The effects of AMF on plant growth markers as seedling height, primary root length, ground diameter, soil physicochemical qualities, and secondary metabolism paclitaxel content were investigated in Taxus chinensis seedlings.   Result  The results showed that: (1) AMF inoculation could significantly promote the growth of plant height, ground diameter, root length, and number of primary branches of Taxus chinensis seedlings, among which the inoculation of R. intraradices and F. mosseae reached a significant level of growth of plant height and root length, and the inoculation of R. intraradices had the most significant effect on the growth of ground diameter, and the inoculation of F. mosseae had the best influence on the increase of the number of first-order branches. (2) The growth indexes of AMF showed significant correlation with soil physicochemical properties such as soil quick-acting phosphorus content, soil alkaline dissolved nitrogen , and soil quick-acting potassium content. The infestation rate showed a significant negative correlation with the quick-acting phosphorus (p<0.05), and other indexes such as alkaline dissolved nitrogen , quick-acting potassium , and growth indexes all showed a significant correlation.(3) Inoculation with AMF might greatly increase the content of paclitaxel, with R. intraradices having the strongest effect.  Conclusion  By analyzing the impacts of several AM fungi on Taxus chinensis growth and secondary metabolism during the seedling stage, it was discovered that the symbiotic pattern of Rhizophagus intraradices and Taxus chinensis could better promote growth and accumulation of secondary metabolism products.
Pseudomonas in Hippophae rhamnoides Rhizosphere Affecting Growth of Ipomoea aquatica
GAO Pei, XU Shuqin, HE Xi, SAN Hongyuan, MA Yuhua, YE Guisheng
[Abstract](60) [FullText HTML](35) [PDF 1664KB](3)
Abstract:
  Objective  Pseudomonas sp. in the rhizosphere of Hippophae rhamnoides were isolated and studied for potential application as a biofertilizer.  Method  Microorganisms in the rhizosphere soil of Hippophae rhamnoides sinensis subsp. in the wild in Qinghai Province were isolated by using selected media and purified by plate streaking. Candidate Pseudomonas strains were morphologically, physiologically, and biochemically identified as well as 16S rDNA sequenced. Abilities of the isolates to degrade organic and inorganic phosphorus, potassium, cellulose and/or to fix nitrogen were examined. Effects of spraying the bacterial culture broth of the individual isolates on the seed germination and seedling growth of Ipomoea aquatica Forssk were observed.   Result   On different media of specific formulations, the diameters of the turbid circles born by the 7 isolated Pseudomonas strains cultured for 3d ranged 4.28-13.71mm with 5.15-25.41 μg·mL-1 of dissolved organic phosphorus, those of clear circles 3.51-7.62mm with 2.15-22.26 μg·mL−1 of dissolved inorganic phosphorus, those of halos 11.12-21.85mm with 5.07-14.33 μg·mL-1 of dissolved potassium, those of transparent circles 4.61-10.22 mm of cellulose-degradation, and the ratios of the nitrogen-fixing clear circle diameter (D) to the colony growth circle diameter (d) 1.33-1.86. The isolated Pseudomonas strains significantly improved the I. aquatica seed germination rate and seedling growth. Among them, ZGSJ-3 showed the largest increases of 35.2% on the 3.69 mm leaf width and of 41.2% on the 50.25 mm stem length over control.   Conclusion  Presence of the Pseudomonas sp., especially ZGSJ-3 and ZGSJ-7, isolated in this study significantly improved the seed germination and seedling growth of I. aquatica.
Study on matrix effects in the detection of 15 pesticide residues in 6 types of tea leaves
CHEN Lu, LI Huaiping, WANG Guxi, TANG Xiangkai, FENG Dejian, WEI Yuting
[Abstract](103) [FullText HTML](42) [PDF 702KB](6)
Abstract:
  Objective  Matrix effects in detecting 15 pesticide residues in six types of tea using gas chromatography-tandem mass spectrometry (GC-MS/MS) with a sample pretreatment of QuEChERS were analyzed for assay improvement.   Methods  Tea samples were pretreated with QuEChERS, and solvent and matrix standard curves for the mass concentrations of 0.01-0.32 mg·L−1 prepared. After a GC-MS/MS determination, the matrix effects were calculated.   Results  Matrix effects were enhanced in the determination that ranged from 1.35-411.58% with the strong, medium, and weak matrix effects of 73.33%, 23.33%, and 3.33%, respectively. When black tea was used as an alternative matrix, they ranged 0.62-59.22% with the strong, medium, and weak matrix effects at 1.33%, 14.67% and 84.00%, respectively. Whereas, when a mixed tea was applied as an alternative matrix, suppressed matrix effects resulted at 0.09-48.09% with the strong, medium, and weak matrix effects at 0, 13.33% and 86.67%.   Conclusion  The matrix effects in the pesticide residue detection on teas were complex. Application of a matrix standard curve reduced the matrix effects and improved the determination accuracy. By using mixed tea sample as an alternative matrix, the analysis efficiency was significantly enhanced.
Analysis of combining Ability and Correlation on Yield, Brix and Three-ear-leaves Characters of waxy maize
CUI Yang, LIU Junfeng, ZHANG Yao, SONG Qiaoheng, KONG Liangliang
[Abstract](162) [FullText HTML](74) [PDF 547KB](8)
Abstract:
:   Objective  To study the genetic rule of waxy maize yield and sugar degree, and analyze the correlation between yield, sugar degree and three ear leaves traits, so as to provide reference for breeding of fresh waxy maize with high sugar degree and explore the mechanism of the influence of three ear leaves traits on quality traits.   Methods  In this study, 6 inbred lines were used as test species, and 15 waxy maize inbred lines were selected for incomplete diallel hybridization (NCII design). The yield of ear, soluble sugar content (sugar degree) of grain, leaf length, leaf width, leaf area, and other 10 ear and plant-related traits of the hybrid combinations were measured at the harvest period (21 days after pollination). Panicle weight and sugar content were the main yield and quality traits to evaluate the contribution of L. trefoil to yield and quality. The other characters were used as reference characters to evaluate the importance of three ear leave to yield and quality. According to the measured data, general combining ability (GCA) and special combining ability (SCA) of ear yield and sugar content were analyzed.   Results  The results showed that there was a strong correlation between leaf length and ear yield in three ear leave characters. There was a strong correlation between leaf width and sugar content. There was a significant negative correlation between ear yield and sugar content, so materials with moderate leaf length and wide leaf width should be preferred in breeding process to improve the selection efficiency of high-quality waxy maize.   Conclusion  The study indicated that the length and width of the leaves, especially the lower ear leaves and ear position leaves, could be used as the basis for selecting breeding materials of waxy maize.
Effect of Waterlogging on Quality of Maize Kernels at Filling Stage
GONG Xue, LU Shengqiao, CHEN Kun, QIN Hongyu, ZHOU Jinguo, WEI Zhengyi, ZHONG Changsong, YANG Yaojiong, JI Yuqi, LIU Yali, ZHANG Shukuan
[Abstract](350) [FullText HTML](136) [PDF 1123KB](7)
Abstract:
  Objective  Effects of waterlogging on quality of maize kernels at filling stage were studied to decipher the mechanism and for breeding selection.  Method  Two backbone maize inbred lines in Guangxi, 88M-1-8 and Xian 21A, were subjected to normal water irrigation (CK) or artificial flooding treatments (W) after pollination. The experiment lasted 14d or 18d with a total of 4 variables, i.e., CK-14, CK-18, W-14, and W-18. Contents of soluble protein, soluble sugar, starch, sucrose, and abscisic acid (ABA) as well as activities of sucrose synthetase (SS-I in decomposition direction) and granule-bound starch synthase (GBSS) of the kernels were measured. Effect of waterlogging on maize quality at filling stage was evaluated using the principal component analysis and membership function method.   Result   Under W-14, the contents of protein, starch, and ABA and the activities of SS-I and GBSS in 88M-1-8 were significantly higher than those in Xian 21A, so were the soluble sugar, starch, ABA, and GBSS in 88M-1-8 under W-18. As the waterlogging prolonged, the soluble protein, starch, sucrose, SS-I, and GBSS in both inbred lines raised significantly. The protein, starch, ABA, and SS-I in 88M-1-8 increased more significantly, and the soluble sugar, sucrose and GBSS in Xian 21A more significantly in W-14 than CK-14. Furthermore, compared with CK-18, W-18 induced more significant increases on the soluble sugar and ABA in 88M-1-8, and the protein, starch, sucrose, SS-I, and GBSS in Xian 21A. Evaluated by the principal component analysis and membership function method on various kernel quality indicators, the tolerance of the two species to flooding were 88M-1-8 under W-14>88M-1-8 under W-18>Xian 21A under W-14>Xian 21A under W-18.  Conclusion   Artificially flooding the maize plants raised the soluble protein, soluble sugar, starch, sucrose, SS-I, and GBSS in kernels over regular irrigation. 88M-1-8 and Xian 21A differed in responses to the stress. Waterlogging-tolerant 88M-1-8 was significantly higher in the GBSS activity and ABA and starch contents than Xian 21A. Nonetheless, prolonged waterlogging reduced the stress tolerance of either cultivar.
Changes of Microbial Community in Agaricus bisporus Culture Substrate during Fermentation
YUAN Bin, KE Lina, LIAN Yanping, ZHAO Guanghui, FENG Weilin, CAI Weiming
 doi: 10.19303/j.issn.1008-0384.2022.009.018
[Abstract](463) [FullText HTML](553) [PDF 2371KB](105)
Abstract:
  Objective  Changes of the microbial community in fermentation of substrates for cultivating Agaricus bisporus were investigated.   Method  The microbial community characteristics of a compound A. bisporus culture substrates containing spent Flammulina velutiper and Pleurotus eryngii materials were monitored during the 7 stages, Ag1 to Ag7 from pile building to 1st and 2nd fermentation, using the high throughput 16S rDNA full-length sequencing of Pacbio platform.   Result  A total of 715 OTUs were obtained in the fermentation process (i.e., 328, 340, 294, 377, 364, 166, and 174 for each stage) with 161 OTUs commonly found in all stage. The microbes included 21 phyla, 299 genera, and 399 species. At phylum level, Fimicutes, Proteobacteria, Bacteroidetes, and Gemmatimonadetes had higher abundance in all 7 stages. The dominant genus in the substrate during the pile building and 1st fermentation stages was Ureibacillus, while Limnochordaceae, S0134_terrestrial_group, Thermobacillu, and Ruminiclostridium stood out in the 2nd fermentation stage. At species level, Ureibacillus thermophilus and Ureibacillus terrenus were dominant in the pile building and 1st fermentation stages. In the 2nd fermentation stage, the species in the genus of Limnochordaceae were the greatest in relative abundance. It appeared that the microbial abundance and diversity increased gradually before the onset of 2nd fermentation. The community structure of the substrate varied significantly between the 1st and the 2nd fermentation, but the deviation diminished significantly after the 2nd fermentation leaving mostly the dominant species that basically functioned to degrade the waste mushroom material enriching the fertilization effect.   Conclusion  The full-length sequencing technology clearly identified the dominant species unveiling many previously unclassified microorganisms. The results helped to better optimize the substrate fermentation process for an improved cultivation of A. bisporus.
Effects of coffee waste mulching on photosynthesis and water use efficiency of coffee seedlings
ZHANG Ang, DONG Yunping, LIN Xingjun, ZHAO Qingyun, SUN Yan, LONG Yuzhou, ZHONG Yiming, TAN Jun
[Abstract](431) [FullText HTML](154) [PDF 946KB](14)
Abstract:
  Objective  The effects of coffee peel and litter on the growth and photosynthesis of coffee seedlings were investigated to determine a suitable ecological cycle cultivation a suitable of coffee.   Method  One-year-old seedlings of seedlings were used as experimental materials. A randomized block design was used to study the effects of coffee peel and litter on their leaf photosynthesis and water use efficiency. Conventional mulching group was set up: C (control), L (litter mulching cultivation), P (peel mulching cultivation) and LP (litter and peel mulching cultivation).   Result  Coffee litter mulching significantly increased the specific leaf area of coffee by 45.46%, while coffee peel mulching significantly decreased the plant height by 12.11%. Coffee peel mulching significantly increased net photosynthesis, leaf respiration, total photosynthesis, net water use efficiency and total water use efficiency by 78.33%, 109.34%, 91.72%, 80.54% and 104.95%, but did not affect stomatal conductance, transpiration rate and carbon use efficiency. The comprehensive evaluation of coffee photosynthesis under coffee waste mulching treatment was P>LP>L>C.   Conclusion  Coffee peel mulching was better than other mulching treatments in promoting the growth and photosynthetic capacity of coffee seedlings. The suitable coffee ecological cycle cultivation with the peel amount of 239.05 g m-2 could to realize cost saving and efficiency increase in coffee planting process.
Carbon, Nitrogen, and Enzyme Activities in Saline-alkali Soil on Songnen Plain as Affected by Land Uses
LIU Qian, GUO Boya, WU Xiuyu, WANG Yue
[Abstract](404) [FullText HTML](235) [PDF 992KB](7)
Abstract:
  Objective  Fertility and enzymatic activity of the saline-alkali soil in relation to land use were analyzed for ecological improvements and restoration.   Method  At Songnen Plain in western Jilin province, the effects on organic carbon, total nitrogen, invertase, urease, alkaline phosphatase, and catalase of the saline-alkali soils under different types of land use as paddy farming field (N1), dry farming field (N2), wetland (S), and grassland (C) were compared.   Result  The organic carbon contents in the soils ranged 9.70–16.27 g·kg1 under N1, 3.85–11.58 g·kg1 under N2, 2.14–2.97 g·kg1 under S, and 5.25–11.24 g·kg1 under C; and the total nitrogen, 1.83–2.32 g·kg1 under N1, 0.45–0.76 g·kg1 under N2, 0.34–1.28 g·kg1 under S, and 0.88–2.04 g·kg1 under C. The activities of various enzymes were urease>alkaline phosphatase>catalase>sucrase and decreased along the depth of the soil layers. The invertase significantly correlated with C/N at P<0.05, the urease with C/N at P<0.01, the alkaline phosphatase with the organic C at P<0.01 and with the total nitrogen at P<0.05, while the catalase with total nitrogen at P<0.01 and with C/N at P<0.05. The redundant analysis indicated that the activities of invertase and urease were mainly regulated by the pH and bulk density, while those of alkaline phosphatase and catalase largely affected by the moisture content and electric conductivity of the soil.   Conclusion  Land use exerted significant effects on the organic carbon, total nitrogen, and enzyme activity in the saline-alkali soils which gradually decreased from the surface to the deeper layers. Farming on the land fostered the nutrient accumulation and increased the enzymatic activities in soil. Thus, either paddy or dry field was more ecologically friendly than wetland or grassland for the regions of saline-alkali soil.
Role of pAPN and NEU3 in TGEV Infection on Pig
LI Zhaolong, FENG Zhihua, ZHANG Bingchen, FANG Zhou, LIANG Wangwang, CHEN Wenzhi
[Abstract](865) [FullText HTML](297) [PDF 1905KB](30)
Abstract:
  Objective  Role of aminopeptidase gene pAPN and sialic acid neuraminidase gene NEU3 in the transmissible gastroenteritis virus (TGEV) infection on pigs was investigated.   Methods  Being the main receptor of TGEV, pAPN was removed from pAPN and NEU3 in ST cells to verify its supposed key function on the disease. The CRISPR gene editing technique was applied to clip the target gene in ST cells prior to an artificial TGEV infection test. The resulting changes on the infection, virus copy number, cytopathic improvement, and fibronectin were monitored.   Results  Compared with control, the ST cells free of pAPN and NEU3 significantly attenuated TGEV infection-induced cytopathies and the virus copy number. In addition, at a same TGEV titer the mRNA immune responders induced by the knockdown ST cells were significantly lower than the wild-type counterparts.   Conclusion  It was confirmed that the removal of pAPN and NEU3 inhibited the TGEV infection in pigs with reduced viral induced cytopathies. Thus, an antiviral therapy and a guideline for breeding resistant pigs could be developed by targeting these two key genes in the ST cells.
Effects of different duration of waterlogging on physiological characteristics and yield of wheat at booting stage
DING Fugong, LU Yifei, JIA Baosen, ZHU Xudong, XIONG Zehao, WANG Chao, HOU Zehao, LIU Yike, ZHU Zhanwang, ZHANG Yingxin, WANG Shuping, FANG Zhengwu
[Abstract](679) [FullText HTML](345) [PDF 941KB](34)
Abstract:
  Objective  The stress of wheat waterlogging is the main abiotic stress factor in wheat production in the middle and lower reaches of the Yangtze River. To study the effects of different duration of waterlogging on physiological characteristics and yield of wheat at booting stage provides a theoretical basis for the research on the mechanism of wheat waterlogging resistance at booting stage and production.  Method  The effects of waterlogging duration on wheat growth and yield at booting stage were studied by pot pot water control method with wheat varieties Yangmai 16 and Zhongmai 895 as experimental materials.  Result  (1) Under the stress of waterlogging, the chlorophyll content of wheat leaves decreased significantly. The longer the waterlogging lasted, the greater the decline of SPAD value was. The SPAD value of the more heavily injured leaves decreased more, and the more severely injured the inverted two leaves than the flag leaves. (2) The activity of antioxidant enzymes such as CAT, SOD and POD in wheat showed a trend of type "∧" during the waterlogging period, while the content of reactive oxygen species (ROS) decreased or increased slowly in the early stage of waterlogging, while increased sharply in the late stage. (3) In the booting period, the effective number of ears, grain number of ears, 1000-grain weight and other yield factors increased slightly, which may be caused by the stress response of wheat. (4) The stress of waterlogging at the stage of heading had no significant effect on the height of wheat plant. Long-term waterlogging resulted in a significant decrease in wheat yield. The decrease of effective panicle number, grain number per panicle and 1000-grain weight was the main factor causing wheat yield reduction. After 15 d of waterlogging stress, the yield per plant of zhongmai 895 and yangmai 16 decreased by 51.47% and 43.99%, respectively, compared with CK.  Conclusion  Booting stage waterlogging stress significantly reduced the wheat leaf chlorophyll content, destroyed the plant active oxygen metabolism in the body and the balance between antioxidant enzyme system, excessive accumulation of reactive oxygen species causes cells to peroxide lipid membrane, causing cell structure and function is impaired, affect plant photosynthesis and nutrient transfer and accumulation, increase the biomass of wheat is reduced, resulting in lack of grain-filling, caused empty grain, grain and invalid number of flat significantly increased, resulting in wheat production. In addition, during the whole process of waterlogging stress, the resistance of the two wheat varieties tested was as follows: yangmai 16>zhongmai 895.
Effect of endogenous hormone content and gene expression related to hormone synthesis on tuber expansion of Bikeqi yam
AO Lan-ji-ya, JI Xiang, SHAO Ying, ZHAO Lingmin, ZHANG Yan-fang, HUO Xiu-wen
[Abstract](1008) [FullText HTML](725) [PDF 693KB](40)
Abstract:
  Objective  To investigate the changes of starch content, reducing sugar content and soluble total sugar content in tubers, the changes of endogenous hormone content in tubers, the morphological indexes, physiological indexes and the analysis of variance of endogenous hormones during the process of tuber expansion. And the correlation between these indicators and endogenous hormones, lay the foundation for further study of the physiological mechanism of yam growth and development and guiding the high yield and quality cultivation of yam.  Method  Six kinds of endogenous hormones such as ABA, GA3, IAA, JA, ZR and IPA were determined by enzyme-linked immunosorbent assay with different tuber lengths of Bikeqi yam. High performance liquid chromatography (HPLC) was used to determine the content of salicylic acid (SA), and to analyze the dynamic changes of source hormone content in different growth stages, and the correlation between morphological indicators of yam and endogenous hormones. The relationship between differentially expressed genes associated with endogenous hormones and endogenous hormones was explored for the effects of these factors on the expansion of yam tubers.  Result  The contents of IAA, ZR, ABA, JA and SA were positively correlated with the morphological indexes of yam tuber; the contents of GA3 and IPA were negatively correlated with the morphological indexes; the contents of IAA were positively correlated with the perimeter and diameter of yam tuber; the contents of GA3 were negatively correlated with the length of tuber; the genes correlated with IAA were negatively correlated with the contents of IAA  Conclusion  Endogenous hormones IAA, ZR, ABA, JA and SA promote the expansion of yam tuber; endogenous hormones GA3 and IPA inhibit the growth of yam tuber; endogenous hormones IAA promote the thickening of yam; endogenous hormones GA3 inhibit the elongation of yam; the down-regulation of IAA related genes can promote the synthesis of IAA, that is, regulate the content of IAA.
2024, 39(8): 1-2.  
[Abstract](31) [FullText HTML](16) [PDF 369KB](64)
Abstract:
Animal Science
Construction and Biofilm Formation of yibT-and-csgD-deleted Salmonella typhimurium
LIU Xiaotong, WANG Bingbing, WANG Zhimei, LIU Feifan, ZHAO Wei, QI Yonghua
2024, 39(8): 879-887.   doi: 10.19303/j.issn.1008-0384.2024.08.001
[Abstract](54) [FullText HTML](29) [PDF 1048KB](60)
Abstract:
  Objective   Strains of Salmonella typhimurium with yibT and/or csgD deleted were created to observe the effect on its biofilm formation for effective prevention and control of the foodborne pathogen.   Method   The λ-red homologous recombination technique was applied to create the gene-deleted mutants, WTΔyibTΔcsgD and WTΔcsgD, of WT S. typhimurium (CVCC541). Subsequently, the gene-complemented strains were also obtained using recombinant vector technology. Differences of the mutants in biofilm formation were determined by crystal violet staining, content of exopolysaccharide measured by phenol-sulfuric acid, mobility tested on semi-solid agar plates, and self-aggregation monitored. Under a scanning electron microscope, biofilm structure was observed. Finally, mRNA expressions of the target genes in the biofilm were identified by fluorescence quantitative PCR.  Result  A double-gene deleted WTΔyibTΔcsgD, a WTΔcsgD without csgD, and their respective complemented strains, WTΔcsgDΔyibT/pcsgD and WTΔcsgD/pcsgD, were successfully obtained. The double-gene deletion significantly reduced the ability in forming biofilm, the content of exopolysaccharide, and the aggregation of the genes to targets, and the mRNA expressions of invF and sdiA in WT S. typhimurium.   Conclusion  Deletion of yibT and csgD significantly impacted some critical biofunctions of S. typhimurium that could become a new approach for control of the foodborne disease.
Preparation and In Vitro Inhibitory Effects of Specific Tripple Anti-vibrio Egg Yolk Antibody
ZHANG Linpeng, HONG Yucong, SUN Kaihui, ZHAO Shuyan, LIU Lihan, LIN Li, QIN Zhendong
2024, 39(8): 888-897.   doi: 10.19303/j.issn.1008-0384.2024.08.002
[Abstract](45) [FullText HTML](25) [PDF 1647KB](61)
Abstract:
  Objective   In vitro inhibitory effects of the prepared specific anti-vibrio antibody on Vibrio vulnificus, V. fluvialis, and V. cholerae were studied for the development of a bioagent to prevent and treat infections caused by the viruses.   Method  Trivalent inactivated vaccine against the three specific viruses were prepared to immunize hens, and eggs collected for producing the specific IgY. Potency, purity, and specificity of the IgY were determined. Antiviral effects of the antibody were tested by immunofluorescence electron microscopy, scanning electron microscopy, and an in vitro test.   Result   Indirect ELISA detected the specific IgY with peak titers as high as 1:51200 that sustained for 4–5 weeks when the target viruses were used as antigens. The SDS-PAGE analysis on the IgY purified by means of water dilution and salt precipitation showed a gradual rise in purity, reduction in protein impurities, and increase in visible heavy and light bands. The immunoblotting of the specific IgY showed its binding to the corresponding antigens with appearance of the protein bands. The immunofluorescence electron microscope displayed apparent agglutination on the bindings to the three viruses. And furthermore, a scanning electron microscopic image of the viruses treated with the specific IgY detailed the adhesion of the antibody to and structural disruption of the virus cells. Finally, significant in vitro, dose-dependent inhibitory effects of the specific IgY on the growth and proliferation of target vibrio strains were observed.   Conclusion  The prepared specific triple anti-vibrio egg yolk antibody exhibited significant in vitro inhibitory effects on V. vulnificus, V. fluvialis, and V. cholerae. It seemed plausible that the application could become an effective alternative to antibiotic treatment in preventing and controlling the diseases caused by these viruses.
Identification and VP1 Sequencing of Duck Hepatitis A Virus Type 3
JIAO Wenlong, FU Qiuling, LIN Yongqiang, LIU Yousheng, LIU Rongchang, LIANG Qizhang, JIANG Nansong, WAN Chunhe, CHENG Longfei, CHEN Hongmei, LIN Jiansheng, FU Guanghua, HUANG Yu
2024, 39(8): 898-905.   doi: 10.19303/j.issn.1008-0384.2024.08.003
[Abstract](46) [FullText HTML](19) [PDF 1191KB](58)
Abstract:
  Objective  Pathogen that caused duck liver hemorrhage in Anhui Province was identified and its genetics studied.   Method  A suspected virus strain was isolated from the ducks suffered from liver hemorrhage at a poultry farm in Anhui Province. Pathogenic nucleic acid and an animal regression test were employed to identify the culprit. VP1 of the confirmed duck hepatitis A virus type 3 (DHAV-3) were sequenced and analyzed using biological software.  Results  No bacterial pathogens were isolated from the culture of the diseased duck liver tissue. However, the specimens were tested positive for DHAV-3 but free of other viruses commonly known for duck liver hemorrhages by RT-PCR. The duck embryos inoculated with the isolate died with massive hemorrhages, and the 5th generation allantoic fluid detected presence of DHAV-3 by RT-PCR. The isolate was subsequently code-named AH230225 and determined to have the effective lethal dose 50 (ELD50) of 10−4.17/0.1 mL. In an animal regression test, the Cherry Valley ducklings had a mortality rate of 80%. The dissected lesions in the liver and kidneys of the dead ducks were similar to the typical clinical specimen. The sequenced VP1 nucleotides of AH230225 showed the greatest homology of 98.8% with the DHAV-3 of Anhui isolate, AH07. Its homologies with the 10 strains of DHAV-3 listed in GenBank were 90.4%~98.8%, with DHAV-1, 62.1%–63.0%, and with DHAV-2, 64.6%–64.9%. It appeared that the VP1 of AH230225 was genetically most closely related to that of AH07 but farther from those of SD01, G, or Korean AP-04009 and AP-03337. In other words, it was distant from DHAV-1 and DHAV-2 on the evolutionary branch.  Conclusion  The virus that caused the liver hemorrhage on ducks at the farm in Anhui was identified to be DHAV-3 with VP1 closely related genetically to that of AH07.
Crop Science
Crop Yield, Kernel Sugar, and Three Ear-leaves of Waxy Maize
CUI Yang, SONG Qiaoheng, LIU Junfeng, ZHANG Yao, KONG Liangliang
2024, 39(8): 906-913.   doi: 10.19303/j.issn.1008-0384.2024.08.004
[Abstract](44) [FullText HTML](20) [PDF 572KB](60)
Abstract:
  Objective  Genetic inheritance of crop yield and kernel sugar content relating to measurements of three leaves nearby an ear on a waxy maize plant was examined for correlation to facilitate breeding and quality prediction.  Methods   Six inbred waxy maize lines were cross-bred with 15 key inbred lines based on an incomplete diallel hybridization of NCII design. At harvest, yield of ears, soluble sugar content of kernels, dimensions of the three leaves closest to an ear, and 10 additional traits of the hybrids were measured 21 d after pollination. Ear yield by weight and kernel sweetness by Brix were obtained. Dimensions of the leaves located above, at, and below an ear on a plant were measured to correlate with the general combining ability (GCA) and special combining ability (SCA) on kernel yield and sweetness of the hybrids passed on from their parents.   Results   Correlations were found between the leaf length and the ear yield and between the leaf width and the kernel sugar content. However, the narrow heritability of leaf length and ear yield was relatively low, mostly shown by the SCA on length of lower ear-leaf and weight of corn-on-the-cob. On the other hand, that of leaf width and sugar content was significant and mainly the GCA on width of upper ear-leaf and kernel sugar.   Conclusion   The width of upper ear-leaf grown next to an ear significantly reflected the sweetness of the kernels born on a waxy maize plant. Thus, the measurement could potentially be used as a visual, easily assessable indicator for cultivars selection in breeding and/or quality estimation in forecasting a harvest.
Fungal Community and C, N, P, and S Functional Genes in Rhizosphere Soil of Cassava Field Treated with a Slow-release Fertilizer
ZHAO Xinxin, WEI Yundong, CHEN Ruirui, ZHOU Shiyi, ZHENG Hua, MA Chongxi, XU Chuan, LI Jun, LU Saiqing
2024, 39(8): 914-926.   doi: 10.19303/j.issn.1008-0384.2024.08.005
[Abstract](31) [FullText HTML](19) [PDF 1735KB](58)
Abstract:
  Objective  A slow-release fertilizer was applied on a cassava lot to analyze the responses of the fungal community and C, N, P, and S functional genes in the rhizosphere soil.   Method  A field experiment was conducted with treatments of no fertilization (T1), basal application of double-coating slow-release fertilizer C2 (T2), and C2 applied 34 d after planting (DAP) (T3). Rhizosphere and bulk soil samples were collected at 77, 104, and 134 DAP to determine fungal diversity according to ITS rRNA sequenced by a high-throughput Illumina Miseq PE300, copies of 72 functional genes of C, N, P, and S cycles (including total DNA) by the gene chip technology, and available nutrients by chemical analysis for a correlation analysis.   Result  (1) Significant differences on the relative abundance (RA) of Mortierellomycetes, Tremellomycetes, and Orbiliomycetes were found in the rhizosphere soils on 104 DAP showing T2<T1, while that of Scatterocysts on 134 DAP indicating T2>T1. Fungal class Scatterycetes under T2 on 134 DAP, Rubiaceae under T3 on 77 DAP, and Coprochetes under T3 on 104 DAP were relatively enriched in the rhizosphere than in the bulk soil. The RAs of the rhizosphere fungi also differed significantly on time of sampling and under different treatments. They were 134 DAP<104 DAP for Mortieromycetes under T1, 134 DAP>77 DAP for Scatterocystae and Coprochestae under T2, and 104 DAP>77 DAP for Sphaeromycetes under T3. (2) The Sobs, ACE, and Chao1 indexes under T1 on 104 DAP, T2 on 104 DAP, and T3 on 134 DAP were significantly higher in the rhizosphere than in the bulk soil (P<0.05 or P<0.01). The Shannon index of rhizosphere soil was lower under T1 than under T2 or T3 on 77 DAP. Under T1, the index was 104 DAP>77 DAP; and under T2, it was the opposite. The Simpson indexes ranked in the order of the rhizosphere soil under T1 on 77 DAP>the bulk soil under T1 on 77 DAP>the rhizosphere soil under T1 on 104 DAP>the rhizosphere soil under T2 on 77 DAP>the rhizosphere soil under T3 on 77 DAP. (3) The LEfSe analysis indicated the fertilizer applications enriched one class, one order, and two families of fungi in the rhizosphere soil, whereas the bulk soil was more abundant in two species on 77 DAP, in 3 orders, one family, and one genus on 104 DAP, and in one phylum, one order, one family, and one genus on 134 DAP. Two orders on 104 DAP and one class on 134 DAP were enriched in the rhizosphere soil. (4) On 134 DAP, the 9 functional genes, such as lig in the bulk soil under T1, had significantly more copies than in the rhizosphere soil. In the rhizosphere soil, the RAs of chiA and aclB under T1 on 77 DAP were higher than those on 104 or 134 DAP. (5) AK significantly correlated with 31 functional genes on 104 DAP. Some fungal classes, such as Tremella, Sarcoidales, Claviculaceae, and Sphaeromycetes, significantly correlated with 40, 15, 14, and 9 other functional genes, respectively.   Conclusion   Fertilization by ways of T2 or T3 enriched the diversity and abundance of cassava rhizosphere fungal community. Fertilizer used, application time, and rhizosphere could all significantly affect the fungal community structure and some functional genes in the soil. The correlations might lead to further studies to unveil the intricate ecosystem.
Horticultural Science
Regulating JsTPS Promoters by JsMYB108 and JsMYB305 in Jasminum sambac
ZHANG Yue, HU Li, WAN Chao, HU Shuangling, YIN Qinwan, YUAN Yuan
2024, 39(8): 927-937.   doi: 10.19303/j.issn.1008-0384.2024.08.006
[Abstract](36) [FullText HTML](17) [PDF 2049KB](60)
Abstract:
  Objective   Regulation functions of JsMYB108 and JsMYB305 on the promoters of three terpene synthase genes (TPSs) relating to the aroma synthesis of jasmine were analyzed.   Method   The promoter fragments of JsTPSs were cloned by genome walking with the DNA of jasmine leaves as template to determine the sequences of the cis-acting elements in them. The fragments were constructed separately in the reporter vector pGWB433. Then, tobacco leaves were transformed with the reporter vector alone or with pK7FWG2.0-JsMYB108 and pK7FWG2.0-JsMYB305 effect vectors to detect the activation of JsMYB108 and JsMYB305 on the promoters. Binding of JsMYB108 and JsMYB305 to the promoters was verified by the yeast one-hybrid assay.   Result   The cloned promoter fragments of the three JsTPSs were 1 357 bp, 1 849 bp, and 1 005 bp with MYB recognition sites. The elements relating to light response, damage response, and abscisic acid induced cis-acting were predicted in different promoter sequences. The GUS staining and activity detection in tobacco leaves confirmed varying degrees of activity of the fragments by introducing JsMYB108 and JsMYB305. Comparing to control, JsMYB108 expanded the activities 1.96-fold on the promoter of JsTPS1, 6.47-fold on that of JsTPS3, and 4.15-fold on that of JsTPS4, while JsMYB305 did 1.57-fold, 15.18-fold, and 3.12-fold, respectively. The yeast one-hybrid assay further verified the bindings of JsMYB108 and JsMYB305 to JsTPS1, JsTPS3, and JsTPS4.  Conclusion   JsMYB108 and JsMYB305 could activate the promoters of JsTPS1, JsTPS3, andJsTPS4. These two transcription factors might play a key role in the synthesis and metabolism of aromatic terpenes in jasmine flowers.
Transcriptome of Passiflora edulis Leaves under Cadmium Stress
ZHANG Lijie, ZHANG Xiaoyan, XIE Lixue, WANG Jianchao, HUANG Jinghao, LI Tao
2024, 39(8): 938-945.   doi: 10.19303/j.issn.1008-0384.2024.08.007
[Abstract](41) [FullText HTML](20) [PDF 1698KB](58)
Abstract:
  Objective   Molecular mechanism of Passiflora edulis leaves in respond to cadmium (Cd) stress was analyzed for genetic improvement on resisting the heavy metal absorption of the plant.   Method  Transcriptome of the leaves of passion fruit plants under a moderate Cd stress of 50 μmol·L−1 for varied durations in a hydroponic experiment was sequenced. qRT-PCR was applied to verify and analyze the results.  Results  There were 3,465, 1,262, and 2,039 expressed genes in the samples under the Cd stress for 24, 48, and 72h, respectively. Sixty-three specific transcription factors and proteins that responded to the stress were obtained. The transcriptome sequencing was confirmed highly reliable by a qRT-PCR analysis. The GO functional enrichment and KEGG metabolic pathways analyses on the transcriptions showed the differential expression genes (DEGs) to be mainly associated with cell structure, catalytic activity, and transcriptional regulations as well as the photosynthesis and carbohydrate metabolism pathways.  Conclusion  The DEGs in passion fruit leaves responding to Cd stress were mainly enriched in the metabolic and biosynthesis-related pathways. The information would pave the way to studies on curtailing Cd contamination that seriously affects the quality and commercial value of the fruit.
Effects of Abscisic Acid Spray on Hormone Metabolism of Camellia reticulata Seedlings under Drought Stress
WANG Weiqinlan, LIN Xianyu, XIAO Lin, ZHOU Lin, FU Xian, QU Yan
2024, 39(8): 946-958.   doi: 10.19303/j.issn.1008-0384.2024.08.008
[Abstract](33) [FullText HTML](20) [PDF 2697KB](56)
Abstract:
  Objective  Effects of ABA spray on endogenous hormones in Camellia reticulata seedlings under drought stress were studied.   Methods   An ABA solution in the concentration of 100 mg·L−1 was sprayed on the leaves of 2-year-old C. reticulata Zipao seedlings grown in potting soil under a simulated drought stress using PEG_6000. Osmoregulatory substances and hormone metabolome in the roots and leaves of the plants were measured under the treatment and subsequent rehydration.   Results  Osmotic regulation of the seedlings took place mainly in the leaves. Under drought stress, ABA, JA , SA ,IAA, and CKs accumulated in the leaves, while ABA, GA , and CKs increased but IAA decreased in the roots. Upon rehydration, ABA, GA, and CKs in the roots gradually declined, while IAA rose, meanwhile, the leaves started regulating with the stored JA, SA, IAA, and CKs. The KEGG enrichment analysis showed that the diterpene biosynthesis pathway in the roots and the hormone signal transduction and zein biosynthesis pathways in the leaves were significantly enhanced. There was a significant correlation between the osmoregulatory substances and endogenous hormones. The ABA spray boosted the contents of soluble protein, soluble sugar, ABA, GA, and CKs in the roots as well as those of free proline, ABA, and JA in the leaves improving the drought tolerance of the plant. With replenished water, CKs in the roots and JA in the leaves were reduced to encourage IAA synthesis, which in turn, aided the recovery of plant functions.  Conclusion   The responses of the roots and leaves of C. reticulata seedlings to the imposed drought and the subsequent rehydration were different. Application of foliar ABA spray could improve the draught resistance of the plants.
Changes in Morphological Characteristics and Endogenous Hormone Content during Flower Bud Differentiation in Bombax ceiba
HU Shuangling, LIN Gongtian, CHEN Zhehong, YIN Qinwan, CHEN Qingxi, YUAN Yuan
2024, 39(8): 959-967.   doi: 10.19303/j.issn.1008-0384.2024.08.009
[Abstract](40) [FullText HTML](22) [PDF 2195KB](58)
Abstract:
  Objective  Morphology and endogenous hormones of Bombax ceiba flower buds in differentiation were studied to aid the control of fallen debris at fruiting stage of the popular landscape plant in southern China.   Methods   Morphology of the flower buds of B. ceiba at differentiation phases were observed with the paraffin sections under a microscope and contents of endogenous GA3, IAA, ABA, and ZR determined.   Results  The flower bud development was classified in the phases of pre-differentiation, primordium differentiation, sepal primordium differentiation, petal primordium differentiation, stamen and pistil primordium differentiation, and stamen and pistil formation. The flower primordium differentiated in early October, while the sepal primordium did in mid-October, the petal primordium in late October, the female and stamen primordium in mid-early November, and the female and stamen formed in late November. During differentiation, GA3 and ABA in the buds were significantly higher than IAA and ZR. They generally decreased, but IAA declined at first, followed by an increase and another decline, whereas ZR rose initially and fell subsequently. The ratios of ZR/GA3 and (ZR+IAA)/GA3 were on an increasing trend, GA3/IAA decreasing, ZR/IAA and ABA/IAA rising at first followed by declining, ABA/GA3 rising-falling-rising, and (GA3+ZR+IAA)/ABA falling-rising-falling as the differentiation progressed.   Conclusion  The process of B. ceiba bud differentiation can be divided into six stages based on its morphological structure, in which the contents of ABA and GA3 are much higher than those of ZR and IAA , and the changes in the contents showed a significant negative correlation with the bud differentiation process. Consequently, applying either GA3 on the plant prior to the process begins or ABA at the petal primordial differentiation phase could deter the floral development and mitigate the nuisance of causing human allergy and traffic interference due to the massive tree falloffs.
Appearance and Quality of Zheli No. 6 Pears Affected by Bagging
ZENG Shaomin, CHEN Xiaoming, HUANG Baoping, HUANG Xinzhong, JIANG Cuicui
2024, 39(8): 968-976.   doi: 10.19303/j.issn.1008-0384.2024.08.010
[Abstract](40) [FullText HTML](17) [PDF 1065KB](55)
Abstract:
  Objective  Improvements on the appearance and fruit quality of Zheli No. 6 pear by bagging were studied.  Method  Zheli No. 6 pear s on the trees were covered once or twice with a variety of pouches at different times of the fruit development in a field experiment. Appearance and eating quality of the mature pears were monitored.   Result   The bagged fruits were significantly brighter and more shiny than the counterparts without covering (P<0.05). The pears matured in the white single-layer large pouches had significantly lower fruit rust index (FRI) than control (P<0.05) but no significant differences on the contents of soluble solids, total soluble sugar, and sweetness. Those being bagged twice were significantly lower on FRI than control or those being bagged only once. Nor did they differ significantly from control on the contents of soluble solids, total soluble sugar, titratable acid, and VC, but contained significantly more fructose and glucose than control or those were bagged a single time (P<0.05). When the bagging was held 10d, 20d, and 30d after full bloom, equal to or more than half of the resulting fruits were yellowish green with few rusty streaks and spots having an FRI of 0.471-0.600 and low on latitudinal measurements and flesh density. Bagging 60d after full bloom produced 72.5% of the fruits with fully rusty-appearing peels. And, except for those bagged 10d after full bloom, no bagged pears significantly differed from control on the contents of soluble solids, total soluble sugar, titratable acid, and VC.   Conclusion   Bagging Zheli No. 6 pears on the tree with the white single-layer large pouches significantly affected the appearance of the matured fruits. Using the white small wax pouch in 10-30d after full bloom followed by the white single-layer large pouch for the bagging fostered bearing of yellowish green fruits. Whereas the non-rust-proof pouches for bagging once in 60d after full bloom encouraged formation of a fully rust, brown peel. Meanwhile, the appealing eating quality of Zheli No. 6 pears would basically be intact under the bagging treatments.
Plant Protection
Host Selection of Anagrus nilaparvatae
ZHANG Junian, LIAN Ling, WU Chunzhu, DONG Meng, SHI Longqing, XIE Zhenxing, JIANG Zhaowei, ZHAN Zhixiong
2024, 39(8): 977-983.   doi: 10.19303/j.issn.1008-0384.2024.08.011
[Abstract](28) [FullText HTML](18) [PDF 846KB](56)
Abstract:
  Objective   Reproductive biology of Anagrus nilaparvatae strains (Ans) that lay eggs on different pests was studied to determine the best parasitoid/host matches for biocontrol.  Method  Nilaparvata lugens (Nl),Sogatella furcifera (Sf), Laodelphax striatellus (Ls), and Ishiharodelphax matsuyamensis (Im) were used in an indoor experiment as the hosts to study the reproductive behavior of Ans. Fitness of parasitoid/pest pairings as well as host preference by specific stains of An were determined based upon the size, body color, egg load, parasitism capacity, larva emergence, sex ratio, and lifespan of the wasps.   Result   The body lengths of different species of adult Ans varied significantly, as it was 728.03 μm for those hosted on Nl (AnNl), 715.5 μm on Sf (AnSf), 633.17 μm on Ls (AnLs), and 509.1 μm on Im(AnIm) . The color of wasp body also varied depending upon the pests they chose to lay eggs on. For instance, AnNl, AnSf, and AnLs were mostly light orange, with a few yellowish green, in color, but all AnIm had dark reddish orange bodies. Significant differences in fecundity existed among the strains as well. The egg counts per female were similar between AnNl at 32.93 and AnSf at 32.97 but 27.47 for AnLs and 11.83 for AnIm. The parasitism of these wasps ranked AnNl or AnSfAnLsAnIm. On the other hand, no significant differences were detected on the rate of larvae emerged from eggs, male/female ratio, or lifespan of them. Nor was on host selection—they simply deposited eggs randomly based on availability. The intrinsic rate of increase (rm) of AnNl was the highest at 0.1531 followed by 0.1482 of AnSf, 0.1307 of AnLs, and 0.0427 of AnIm.   Conclusion   The strains of An that parasited on Nl and Sf had significantly greater egg load, parasitism capability, and rm than those did on Ls or Im . Hence, they could be the better candidates to be artificially propagated for biocontrol on the pests.
Resources and Environmental Science
Physicochemical Properties and Microbial Community of Soil and Crop Yield under Rice-Tobacco-Milk Vetch Rotation Cropping
WU Tianyi, GU Gang, ZHANG Binghui, ZHANG Zongping, CHI Guosheng, WANG Ruiqi, LI Tong, CHEN Rui, XIE Xiaofang
2024, 39(8): 984-992.   doi: 10.19303/j.issn.1008-0384.2024.08.012
[Abstract](40) [FullText HTML](18) [PDF 1178KB](57)
Abstract:
  Objective   Regulatory functions of milk vetch on soil fertility and microbial communities were studied to determine the potential of incorporating the shrub plant in rotation cropping with tobacco and rice for further land use improvement.   Method   Soil samples were collected from a field practicing tobacco-rice-milk vetch rotation cropping for 5 years and one of tobacco-rice as control. Physicochemical analysis on the soil using spectrophotometry and metagenomic sequencing on the microbial community were conducted.   Result   With milk vetch added to the tobacco-rice rotation cropping, the field soil increased significantly on the available nitrogen and phosphorus, rose slightly on the organic matter, total nitrogen, and total potassium, maintained a same level of total phosphorus, and reduced significantly on the available potassium. The yield of tobacco rose 2.74% and that of rice 4.67% in 2023. And the microbial diversity became significantly enriched by 8.67% and 3.10% but declined by 11.57% over control on the dominant kingdoms of Proteobacteria, Nitrospira, and Acidobacteria, respectively. The microbes in the soil were largely associated with carbohydrate, energy, and amino acid metabolisms.   Conclusion   By incorporating milk vetch in the rotation cropping of tobacco and rice, aside from the increased yields on the crops, the physiochemical properties of field soil were significantly improved as well.
Differentiations between Microbes in Cultivated and Wild Paris Polyphylla Field Soils
SU Hailan, ZHENG Meixia, JIANG Baodong, LIN Fengfang, CHEN Hong, ZHU Yujing, ZHU Yanming, NIU Yuqing
2024, 39(8): 993-1005.   doi: 10.19303/j.issn.1008-0384.2024.08.013
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Abstract:
  Objective  Diversity of the microbial communities in soils of cultivated and wild Paris polyphylla var. chinensis fields were compared.   Method   Total DNA of the microbes on cultivated land grown P. polyphylla plants or at field of the plants in the wild were sequenced using high throughput Illumina Miseq (2×300 bp). Structure and abundance of the microbial communities in soils of the fields were comparatively analyzed by LDA Effect Size.   Result   The microbial diversity of the wild P. polyphylla lots was richer than the cultivated land. The Chao, Ace, Shannon, and Simpson indexes of the wildP. polyphylla soil were higher than the cultivated counterparts. The communities significantly differed on the abundant phyla of Firmicutes, Nitrospirae, and Spirochaetae, and on the genera of Bacillus, Ktedonobacter, and Paenibacillus. LDA Effect Size showed Firmicutes and Nitrospirae to be the predominant phyla, while Bacillus, Paenibacillus, Tumebacillus,Mucilaginibacter, Nitrospira, Shimazuella, and Singulisphaera the dominant genera in the wild. In the cultivated soil, the Armatimonadetes phylum and the Bryobacter, Aquicella, and Ktedonobacter genera predominated the community. pH and total potassium content of soil were the critical factors affecting the diversity of a microbial community.   Conclusion  Cultivation and soil nutrients significantly differentiated the microbial composition at a P. polyphylla field.
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