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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.
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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.
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Objective In order to provide a technical support and basic data for the prevention and control of porcine transmissible gastroenteritis and the DNA vaccine research, the DNA vaccine vector of S and N genes of porcine transmissible gastroenteritis virus was constructed and its immunogenicity was tested. Method The A site and D site from the S gene and N gene was amplified from transmissible gastroenteritis virus and the N gene (alone) and 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 composition, tertiary configuration, subcellular localization and dominant B cell epitope of S(A-D) protein and N protein. The recombinant vectors were transfected into PK-15 cells, and expression distribution of the N gene (alone) and the A and D sites (fusion) was detected by indirect immunofluorescence and confocal detection. The recombinant vaccine vector was used to immunize mice alone or in combination, and the IgG antibody level was detected by indirect ELISA. Result The results showed that the A sites and D sites of S gene and N genes were amplified, and their sizes were 498bp, 606bp and 1149bp, respectively. the nucleic acid vaccine expression vectors p-S(A-D)-His and p-N-His for A and D sites (fusion) and N gene (alone) were constructed. Bioinformatics software predicted that when TGEV infected host cells, N protein was mainly located in the nucleus and mitochondria, S(A-D) protein was mainly located in the cytoplasm and mitochondria, S(A-D) protein had 7 dominant B cell epitopes, and N protein had 8 dominant B cell epitopes. Recombinant vectors p-S(A-D)-His and p-N-His were successfully expressed in PK-15 cells, and S(A-D)-His and N-His were distributed in the nucleus and cytoplasm of PK-15 cells. After immunizing mice with vaccine vector, the immune effect was p-N-His ＞ p-S(A-D)-His + p-N-His ＞ p-S(A-D)-His from high to low. Conclusion In this paper, the DNA vaccine vectors of S and N genes of TGEV were constructed, and strong specific antibodies were generated after immunizing mice, which provided the basic material and basis for the development of nucleic acid vaccine of TGEV.
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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.
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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·kg−1 under N1, 3.85–11.58 g·kg−1 under N2, 2.14–2.97 g·kg−1 under S, and 5.25–11.24 g·kg−1 under C; and the total nitrogen, 1.83–2.32 g·kg−1 under N1, 0.45–0.76 g·kg−1 under N2, 0.34–1.28 g·kg−1 under S, and 0.88–2.04 g·kg−1 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.
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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.
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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.
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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.
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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.
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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.
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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.
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Objective Clematis florida Thunb. var. plena D. Don is used in She medicine "Shi Er Shi Chen". Its roots, rich in active compounds, are used medicinally. This study aims to understand the gene annotation, secondary metabolite pathways, and gene functions of Clematis florida Thunb. var. plena D. Don. The goal is to enrich its transcriptome information and lay the foundation for further screening and identification of key genes involved in the metabolic pathways of its medicinal compounds, as well as supporting molecular breeding efforts. Method In this study, the PacBio single-molecule real-time (SMRT) SMRT sequencing was used to generate the full-length transcriptome data of the roots of Clematis florida Thunb. var. plena D. Don. Functional annotation, gene structure analysis, and mining of the terpenoid biosynthetic pathway were conductedusing bioinfromatics tools on the obtained transcript data. Result A total of 62.21 G polymerase read bases were generated, finally 20540 non-redundant high-quality transcript sequences were successfully identified. Gene function annotation was carried out against seven major databases, including NR, NT, Pfam, COG/KOG, SwissProt, GO andKEGG. A total of 19,909 transcripts were successfully annotated in at least one public database, and 8,888 transcripts were annotated in five common databases (NR, NT, COG/KOG, KEGG, GO). GO annotation results showed that 14,911 transcripts were enriched in 53 terms, including biological processes, cellular components, and molecular functions. A total of 19,701 transcripts wrere annotated to KEGGdatabase and classified to 6 major pathways and 44 sub-pathways, with the largest number of transcripts enriched in metabolic pathways. COG/KOG annotation identified 13,204 transcripts, with general function prediction being the most predominant.. A total of 978 transcription factors, 224 lncRNAs, and 7,167 SSRs were predicted. A total of 48 transcripts (16 key candidate genes) involved in terpenoid backbone biosynthesis were identified. Conclusion This study successfully performed full-length transcriptome sequencing of the roots of Clematis florida Thunb. var. plena D. Don, obtaining comprehensive gene function information for this species. It fills the gap in the gene information of Clematis florida Thunb. var. plena D. Don and provides a reference for further studies on its regulatory network, biological characteristics, related metabolic pathways, signaling pathways, and molecular mechanisms.
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Objective To explore the mechanism of lipid synthesis and accumulation in thin shelled oil palm fruits. Method Thin shelled oil palm fruits from different post harvest stages were selected [fresh fruits freshly harvested 185 days after pollination (T1), harvested 24 hours after harvest (T2), and harvested 36 hours after harvest (T3)]. We used LC-MS/MS and RNA-seq techniques to determine and analyze the dynamic changes of each lipid metabolite and differentially expressed genes in oil palm mesocarp during raging. Result During fruit development, 5 lipid classes, 23 lipid subclasses, and 520 lipid monomer molecules were identified. Aldehyde dehydrogenase (ALDH7A1, ALDH2), monoacylglycerol lipase (MGL), phospholipase A1 (PLA1), and glycerophosphodiester phosphodiesterase (GDPD1) may affect the hydrolysis of phosphatidylcholine (PC), thereby affecting the oxidation of lipids in oil palm flesh; By hydrolyzing glycerophosphate choline (GPC), the content of PC is affected; Lipophosphatase (LPP) promotes the synthesis of phosphates and glycerophospholipids; The expression of chlorophyll may be related to the content of chlorophyll in the matrix. The joint analysis results showed that aldehyde dehydrogenase (ALDH7A1, ALDH2), monoacylglycerol lipase (MGL), and phospholipase A1 (PLA1) were significantly negatively correlated with glycerophospholipids such as diacylglycerol trimethyl homoserine (DGTS), phosphatidic acid (PA), phosphatidylinositol (PI), phosphatidylcholine (PC), phosphatidylglycerol (PG), phosphatidylethanolamine (PE), and significantly positively correlated with palmitic acid; GDPD1, lipophosphate phosphatase (LPP), and digalactosylglycerol synthase (DGD1) are significantly positively correlated with glycerophospholipid substances such as DGTS, PA, PI, PC, PG, PE, and negatively correlated with palmitic acid; MGL monoglyceride (MG) and linoleic acid (LA) showed a highly significant positive correlation, while Cerd showed a significant negative correlation; DGD1 and LPP are significantly negatively correlated with MG and LA, and significantly positively correlated with ceramide (Cer). Conclusion It is speculated that ALDH7A1, ALDH2, PLA1, and MGL may inhibit the synthesis of glycerophospholipids and promote the synthesis of fatty acids such as palmitic acid; DGD1, LPP, and GDP1 may promote the synthesis of glycerophospholipids and inhibit the synthesis of palmitic acid and other fatty acids. This study provides a theoretical basis for the quality breeding of oil palm that is resistant to rancidity.
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Objective To deeply understand the genetic diversity and direct utilization value of the germplasm resources of tea plants (Camellia sinensis) in the Shi Pavilion group in Nan’an, and to provide scientific basis for the innovative utilization of these resources. Methods SNP molecular marker technology was used to analyze the genetic diversity and relationships of 17 germplasm resources from the Shi Pavilion group in Nan’an and 6 representative varieties from Fujian. Furthermore, high-performance liquid chromatography (HPLC) and ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) were employed to determine the content of catechins and free amino acids in 10 representative samples from the Shi Pavilion group. Results Forty-four SNP loci suitable for identifying the Shi Pavilion group tea plants were screened, and an SNP fingerprint profile of the germplasm resources was constructed. The UPGMA evolutionary tree revealed that the 23 samples could be divided into four subgroups, with the Shi Pavilion group tea plants closely related to ‘Rougui’ and ‘Qidan’ from northern Fujian. Additionally, the analysis of amino acids and catechins content showed significant differences (P＜0.05) in the major quality components of the Shi Pavilion group tea plants. The total catechins ranged from 107.56 to 177.60 mg·g−1, and the total free amino acids ranged from 8.46 to 32.66 mg·g−1. Among them, ST2, ST3, ST6, and ST17 had higher contents of EGCG or EGCG3"Me, with ST3 having the highest amino acid content and the lowest catechin bitterness index. Conclusion The germplasm resources of tea plants in the Shi Pavilion group in Nan’an are abundant and closely related to tea plant resources in northern Fujian. ST2, ST3, ST6, and ST17 have higher contents of EGCG or EGCG3"Me, and ST3 is suitable for making green tea and can be further developed through breeding programs.
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Objective Excavating the important genes that may be involved in the morphological changes of Agaricus bisporus colony can provide scientific basis for the evaluation of germplasm resources and the breeding of new varieties. Methods As2796, the main cultivated variety of Agaricus bisporus, was used as the test strain. The strandy and aerial hyphae grown on the same plate were collected. RNA of the samples was extracted for transcriptome sequencing and compared with the reference genome sequence of Agaricus bisporus. Results Through transcriptome analysis, a total of 965 differentially expressed genes (DEGs) were identified in A601 (As2796 aerial group) and A602 (As2796 strandy group). GO enrichment analysis results showed that hydrophobin-related DEGs were enriched in fungal cell wall composition. Among them, AGABI2DRAFT_13655, AGABI2DRAFT_136569, and AGABI2DRAFT_193057 belong to the ABH3 protein gene family. The expression level in aerial hyphae is significantly higher thanstrandy hyphae, which may be related to the production of aerial hyphae. The study also found that the hydrophobic surface binding protein A (HsbA)-related gene AGABI2DRAFT_194662 ,the FPKM expression level of this gene is 0 in aerial hyphae and 10.24 in strandy hyphae. Through further analysis of DEGs, 8 DEGs that may be related to lipase isoenzymes were enriched. The FPKM expression levels of these genes in aerial hyphae were significantly higher than strandy hyphae. The SNP/InDel analysis results showed that the number of SNPs in the As2796 aerial group was 110601, in strandy group was 111188. The main changes in single nucleotides were C:G＞T:A, and a total of 2374 common InDels were screened. Conclusion This study successfully obtained transcriptome data on mycelial morphological changes in Agaricus bisporus and unearthed potential related genes and SNP sites, which provides a reference for the evaluation of Agaricus bisporus germplasm resources and the breeding of new varieties..
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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 periods, 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, total 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, total 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 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 upwardand, 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.
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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.
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Objective Analysis of disease and insect resistance genes in FunongA, a three-line indica cytoplasmic male sterile line, will provide an important theoretical basis for the application of the sterile line in breeding production. Method Genomic DNA of FunongA, FudaoB and Huahangsimiao was extracted by CTAB method, then blast resistance genes were detected with molecular markers. Resistance genes related to bacterial blight, viral diseases and brown planthopper were analyzed by high density chip detection. Blast resistance genes of FunongA, Huahangsimiao and FudaoB were obtained by PCR, then sequencing and comparative analysis were performed. The samples were collected at 0h, 24h, 48h, 72h and 96h after inoculation with blast fungus. Total RNA was extracted using the Trizol method, and the expressions of blast resistance genes analyzed by SYBR Green I fluorescent quantitative PCR (qRT-PCR). Result FunongA, FudaoB and Huahangsimiao contain blast resistance genes Pib, Pia, Pid3, Pid2, bacterial blight resistance gene Xa21 and yellow mottle virus resistance gene Rymv1. Furthermore, FunongA and Huahangsimiao contain blast resistance gene Pi5 and stripe virus resistance gene STV11, and FudaoB contain blast resistance genes Pita and Pi37. In addition, they do not contain brown planthopper resistance genes. The fragments of Pi5, Pia, Pib, Pid3, Pid2 were 5672 bp, 2597 bp, 5532 bp, 2865 bp, 3847 bp, respectively. The sequences of Pi5, Pid3 and Pid2 in FunongA were consistent with that in Huahangsimiao, while there were 2 and 5 bases different from Pid3 and Pid2 of Fudao B, respectively. In Funong A, the expression of rice blast resistance genes Pi5 and Pib was significantly induced by blast fungus, and the highest at 72 h after inoculation. The expression of Pia and Pid3 increased gradually with the prolongation of inoculation time, and the highest at 96 h. The expression of Pid2 increased firstly and then decreased, and the highest at 24 h. Conclusion Three-line indica cytoplasmic male sterile line FunongA contains blast resistance genes Pi5, Pib, Pia, Pid3 and Pid2, which significantly induced by blast fungus. Funong A also contains bacterial blight resistance gene Xa21, yellow mottle virus resistance gene Rymv1 and stripe virus resistance gene STV11.
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: 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.
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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.
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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.
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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.
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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 CSS0011741（4CL）、CSS0002940（DFR）、CSS0015968（DFR）and CSS0010687（ANS）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.
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: Objective To clarify the sensitivity of Colletotrichum gloeosporioides population in Fujian Province to pyraclostrobin and its cross-resistance with other fungicides. Methods The sensitivity of 112 strains of C. gloeosporioides isolated from five regions of Fujian Province in 2023 to pyraclostrobin was determined by mycelial production rate method. The resistant mutants were obtained by fungicide domestication, and the genetic stability, fitness and cross-resistance to four different fungicides of the resistant mutants were determined. Results EC50 values of pyraclostrobin for 112 strains ranged from 0.0682 -1.0309 µg·mL−1, and the coefficient of variation was 15.12. The sensitivity frequency distribution was a continuous unimodal curve, which was consistent with the continuous skewed normal distribution. The average EC50 value of 0.2036 ± 0.1215 µg·mL−1 could be used as the sensitivity baseline of C. gloeosporioides to pyraclostrobin. Four mutants with moderate resistance to pyraclostrobin and one mutant with low resistance to pyraclostrobin were induced from two wild strains of C. gloeosporioides. The mutation frequency was 8.3×10−4 and the resistance traits could be stably inherited. Compared with their parental strains, the resistance mutants had different sensitivity to temperature. The survival fitness of the resistant mutants, such as mycelial growth rate, sporulation ability and pathogenicity, was slightly lower than that of their parental strains. A certain field fitness cost of these resistant mutants maybe for exchange more natural competitive advantage. Pyraclostrobin has no cross-resistance with different types of fungicides such as carbendazim, difenoconazole and prochloraz, except the same type of fungicide picoxystrobin. Conclusion The sensitivity of C. gloeosporioides to pyraclostrobin in Fujian Province was relatively high, but still has a moderate resistance risk. In production, pyraclostrobin can be mixed or alternated with different types of fungicides such as carbendazim, difenoconazole and prochloraz to delay the resistance production of C. gloeosporioides to fungicides.
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Objective This study aims to enhance understanding of the interaction between Phalaenopsis equestris and viruses, providing a theoretical foundation for developing effective prevention and control measures against Phalaenopsis equestris diseases caused by virus. Method Firstly, coat protein (CP) genes of cymbidium mosaic virus (CymMV) and odontoglossum ringspot virus (ORSV) were amplified using RT-PCR. Existence of CymMV and ORSV particles in Phalaenopsis equestris cells were identified via electron microscopy showing viral morphology and size. Secondly, the abundance, length, base preference, and origin of virus-derived vsiRNAs were analyzed by small RNA deep sequencing technology. Result Specific amplification of CymMV and ORSV CP genes was achieved via RT-PCR. Electron microscopy revealed the existence of rod-like CymMV virions (300 nm) and linear ORSV virions (500 nm) in Phalaenopsis equestris cells. Small RNA deep sequencing yielded 7563892 and 6133689 CymMV and ORSV-derived vsiRNAs, respectively, exhibiting universality and specificity in abundance, length, base preference, and sense strand distribution. Conclusion Synergistic infection of CymMV and ORSV in Phalaenopsis equestris was observed. CymMV and ORSV vsiRNAs displayed characteristic patterns in abundance, length, base preferences, and sense strand distribution, providing insights into viruses-Phalaenopsis equestris interactions and informing the development of preventive measures against Phalaenopsis equestris associated diseases.
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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.
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Objective To investigate the feasibility of producing astaxanthin by Phaffia rhodozyma using okara as a nitrogen source and optimize the fermentation process, providing a reference for replacing high-cost nitrogen sources such as peptone and yeast extract commonly used in traditional astaxanthin production by Phaffia rhodozyma and reducing production costs. Methods Okara was used as the organic nitrogen source, and the effects of carbon sources, precursor substances, additional nitrogen sources, vitamins, inorganic salts, and other factors on astaxanthin production were analyzed. The interaction among four influencing factors, namely (NH4)2SO4, vitamin E, glucose, and sucrose, was optimized using the response surface methodology (RSM). Results When wet okara was used as the nitrogen source, glucose was found to be the optimal carbon source for astaxanthin production by Phaffia rhodozyma. A combination of glucose and sucrose promoted increased astaxanthin production. Potassium salts such as KCl, KNO3, and K2HPO4, as well as (NH4)2SO4, VB2, VE, and zeaxanthin, significantly enhanced astaxanthin yield. After performing RSM optimization on the four most influential factors (glucose, sucrose, K2SO4, and vitamin E), the optimal medium formulation for astaxanthin production was obtained as follows: 10% wet okara, 0.22% K2SO4, 0.6% vitamin E, 1.08% glucose, and 1.50% sucrose. The measured astaxanthin yield was 32.46 mg·L−1, which was 2.23 times higher than that obtained using the YM medium. Conclusion Okara can be used as the sole nitrogen source for astaxanthin production by Phaffia rhodozyma through fermentation. After process optimization using RSM, the efficiency of astaxanthin production by Phaffia rhodozyma was significantly improved. This study provides a reference for cost control of nitrogen sources and enhancement of astaxanthin yield in production.
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Objective To establish a fluorescent RPA constant temperature rapid detection method for the novel Muscovy duck parvovirus (N-MDPV), providing a visual rapid detection technology for grassroots. Method This study used the conserved VP3 gene fragment of the novel Muscovy duck parvovirus as the target, and used EXO fluorescent probes to specifically bind to the conserved VP3 gene fragment. Specific RPA amplification primers were designed and the target gene was amplified using recombinant enzyme polymerase amplification technology. A fluorescent type of novel Muscovy duck parvovirus RPA constant temperature rapid detection method was established. Explore and adjust the optimal reaction time and temperature of the reaction system, and test the limit conditions for the specificity and sensitivity of the method. The collected nucleic acid of the disease material was used for detection, and the accuracy of the method was compared with traditional PCR and virus isolation identification methods. Result The optimal reaction temperature for establishing the method in this study is 39 ℃, the optimal reaction time is 30 min, and the lowest sensitivity for nucleic acid detection can reach 10 fg fg·μl−1; The nucleic acid of the novel Muscovy Duck Parvovirus was specifically amplified, but there was no cross reaction between the nucleic acid of duck adenovirus type 3, avian adenovirus type 4, duck circovirus, duck plague virus, duck viral hepatitis virus, duck Tambous virus, and novel duck reovirus, indicating good specificity; The RPA rapid detection method, traditional PCR method, and virus isolation and identification method established in this study were used to detect the nucleic acid samples of 38 collected duck tissue samples. The results showed that the positive rates were 36.8% (14/38), 36.8% (14/38), and 31.6% (12/38), respectively; The samples that tested positive for RPA showed positive results through PCR and virus isolation and identification methods, with a positive coincidence rate of 100%. The positive coincidence rate between RPA detection method and traditional PCR method in clinical sample testing is 100%. Conclusion This method can be well applied to the large-scale clinical sample detection of the novel muscovy duck parvovirus in grassroots areas lacking corresponding detection equipment, providing a technical means for the visual and rapid detection of the novel muscovy duck parvovirus.
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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.
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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.
, Available online , doi: 10.19303/j.issn.1008-0384.2022.009.018
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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.
