2020 Vol. 35, No. 1
Display Method:
2020, 35(1): 1-5.
doi: 10.19303/j.issn.1008-0384.2020.01.001
Abstract:
Objective To improve callus induction and seedling formation rates of the anther culture for an expedited and stable breeding program utilizing blast-resistant Indica-japonica hybrid rice Yunyin as a parent, an optimized medium and conditions for the auxiliary breeding technology in creating new materials and varieties were investigated. Method The young spikes of F1 hybrid of Yunyin and a conventional Indica rice Yuejingsimiao were disinfected and inoculated. From 12 formulations of base media, carbon sources, and 2,4-D concentrations as well as culture conditions, an optimal anther cultivation according to the rates of callus induction, green seedling, seedling yield, and anther cultivation performance was selected. Result The maximum anther cultivation performance using the NB medium was merely 0.52%, whereas, M8 significantly higher at 0.90%–1.92%. The M8 medium with sucrose for carbon source reached 11.32% on the performance at the early stage of callus induction. The use of 1.0 mg·L−1 or 2 mg·L−1 2,4-D in the medium did not differ significantly on the performance, i.e., 1.89% vs 1.92%. A peaked green seedling rate up to 18.3% was achieved when 0.2 cm-sized calluses were used for the differentiation. Conclusion The optimal culture conditions of Yunyin Indica-japonica hybrid anthers were to use calluses inducing on M8 medium with added 50 g·L−1 sucrose and 1–2 mg·L−1 2,4-D, then, when grew to 0.2 cm in size, transferring to a differentiation medium.
2020, 35(1): 6-12.
doi: 10.19303/j.issn.1008-0384.2020.01.002
Abstract:
Objective To transfer the broad-spectrum rice blast resistance gene, Pi9, to the restorer line Shuangkangmingzhan for improving its disease resistance. Method The Sanming dominant male sterile line, a carrier of the Pi9 gene, was used as the intermediate “bridge” for the gene transduction experiment. Using the molecular marker assistance selection method, from each backcross generation the male sterile plants with Pi9 marker genotype were selected to backcross- mate with Shuangkangmingzhan until the self-homozygous Pi9 gene marker in the fertile plant was detected. Then, from the selected plant in the BC4F1 generation of the Shuangkangmingzhan disease-resistant isogenic line, the target sterile gene was obtained. Result Of the 6 improved lines selected, 5 were identified by the field resistance verification with significantly improved resistance to rice blast. The combining abilities and agronomic traits of the lines were compared and analyzed to show that the improved line Y25-1 possessed the best general combining ability with a strong blast resistance. Since its main agronomic traits were similar to those of Shuangkangmingzhan, it could conceivably be considered a surrogate of the disease-resistant Shuangkangmingzhan for breeding purpose. Conclusion For an improved blast resistance on rice, the successful transferring Pi9 gene from Shuangkangmingzhan onto Y25-1 through the use of the Sanming dominant male sterile line as an intermediate “bridge” provided a new venue for the breeding.
2020, 35(1): 13-19.
doi: 10.19303/j.issn.1008-0384.2020.01.003
Abstract:
Objective Genetic relationship of 90 avocado (Persea americana Mill.) germplasms collected from Dehong, Baoshan, Xishuangbanna, Puer, and Honghe in Yunnan, China and Shan and Kachin States of Burma was analyzed based on AFLP molecular markers for breeding reference and resource information. Method Genomic DNAs were extracted from the leaves of the avocado plants following the CTAB method. They were amplified using the sequence-related amplified polymorphism molecular markers to determine their genetic diversity and similarity. Separation of the amplified fragments was performed on 5% denaturing polyacrylamide gels stained with AgNO3 to obtain the “0,1” matrix. The number of polymorphic loci, percentage of polymorphic loci, effective number of alleles, and indices of genetic diversity were estimated by POPGENE version 32. The genetic similarity estimated by NTSYSpc-2.11F was used for UPGMA (unweighted pair group method analysis) and PCA (principal component analysis) to classify the avocado germplasms. Result Eight primers with strong polymorphism and high repeatability were screened from 24 AFLP primers generating a total of 1 165 bands. Of the bands, 1 163 (99.83%) showed polymorphism. On average, the number of effective alleles was 1.294 4; Nei's gene diversity (H), 0.209 5; and Shannon's information index (I), 0.353 0. At the genetic similarity index (GS) of 0.752, the germplasms were classified into 4 groups. A single avocado germplasm, No. 70, collected from Baoshan was placed in Group I. Group II had 22 germplasms; Group III, one germplasm, No. 59, from Xishuangbanna; and Group IV, the 64 remainders. The germplasms in Group IV were further divided into Subgroups A, B and C at GS of 0.763. A similar grouping result was obtained from the cluster analysis on the PCR data applied AFLP markers confirming these germplasms shared a high similarity within a same region. Conclusion The 90 collected avocado germplasms were found to be relatively high in genetic diversity. Among them, No. 59 and 70 stood out as two most unique varieties.
2020, 35(1): 20-27.
doi: 10.19303/j.issn.1008-0384.2020.01.004
Abstract:
Objective Genetic diversity and relationships of 56 species of Anthurium andraeanum were analyzed using the inter-simple sequence repeat (ISSR) molecular marker technology for breeding and improvement of germplasms and resource management. Method For the collected Anthurium varieties, 100 UBC primers were screened for the polymorphic ones. Using the universal primers, an ISSR molecular marker system was constructed. The UPGMA cluster analysis was performed, and the Nei's gene diversity and Shannon's information indices calculated on the PCR amplified products. Result Ten polymorphic primers were obtained by screening the 100 UBC primers, and 99 bands amplified. Of the amplified bands, 95 were polymorphic with a rate of 95.96%. The analysis using POPGENE indicated the Nei’s gene diversity index (H) to be 0.303 6 and the Shannon’s information index (I) 0.458 5. Thus, the genetic diversity among the Anthurium varieties was high. The 56 varieties were classified into 12 groups. Conclusion The ISSR results suggested a significant correlation between the clustered groups and sepal color of the flowering plants and the molecular marker technique applicable for the identification. Among the cultivars, Princess Alexia Violet, Fiorino, Sumi, Caixia, Thunderbird Luxury, Black Queen, and King exhibited unique traits among them with substantial genetic distances. They were considered potential candidates as female parent for breeding new varieties and should be preserved in the germplasm collection.
2020, 35(1): 28-37.
doi: 10.19303/j.issn.1008-0384.2020.01.005
Abstract:
Objective Diversity and distribution of the microbial communities in the specialty teas produced in Fujian were studied. Method Fifteen groups of well-known local teas in the province were collected for an analysis based on the high-throughput sequencing targeting 16S rRNA gene. Result A total of 11 472 OTUs encompassing 37 phyla, 95 classes, 150 orders, 207 families, and 277 genera were isolated from the specimens. In the 3 categories, Dahongpao had the greatest richness on species; the Huacha was the most diversified; and, as oolong teas, Dahongpao and Tieguanyin had a similar microbial structure. In 10 groups of Dahongpao samples, the dominant genera were Lactococcus, Bifidobacterium, and Bacteroides; that in 2 groups of Huacha, Bacteroides; and those in 3 groups of Tieguanyin, Lactococcus and Bifidobacterium. Five of the top 10 dominant genera in the teas were lactic acid bacteria. According to the co-occurrence analysis on network, the lactic acid bacteria exhibited a synergistic growth relationship with most of the other significantly associated species. Conclusion Bacilli and Actinobacteria were the predominant bacteria found in Dahongpao and Tieguanyin, while Clostridia and Bacteroidia in Huacha. A bacterial taxonomy analysis indicated that tea category differentiated the structure of their bacterial communities. Overall, lactic acid bacteria dominantly existed in these teas produced in Fujian.
2020, 35(1): 38-43.
doi: 10.19303/j.issn.1008-0384.2020.01.006
Abstract:
Objective Quality of sections of different age on a medicinal Polygonatum cyrtonema Hua plant was analyzed for processing, commercial classification, and clinic dosage determination. Method According to the 2015 edition of Chinese Pharmacopoeia (Part 1 and Part 4), contents of moisture, ethanol extracts, ashes, and polysaccharides in sections of different ages on P. cyrtonema plants harvested in Liuzhi Special Zone of Guizhou Province were determined. Flavonoids content was measured by colorimetry. Result Among sections of different ages on the plants, the lowest ash content of 1.38% was found on the 2-year-old specimens; the highest alcohol extract of 78.04% on the 2-year-olds; and the highest polysaccharides of 14.45% on the one-year-olds. The polysaccharide contents of the one to 3 years old sections met the pharmacopoeia standard, but the 4- and 5-year-olds did not. The highest content of flavonoids was 1.91% found on 2-year-lod plant sections. The content of ashes inversely correlated with those other functional components. The principal component analysis ranked the order of sections on their contents of the functional substances as 2-year-old>1-year-old>3-year-old>4-year-old>5-year-old. Conclusion For processing into market products, the harvested P. cyrtonema plants could be sorted according to the age of plant sections, such as, the 2-year-olds as Grade 1, 1-year-old as Grade 2, and 3-year-old as Grade 3. For clinical applications, appropriate dosages could be determined according to the grades as well.
2020, 35(1): 44-50.
doi: 10.19303/j.issn.1008-0384.2020.01.007
Abstract:
Objective To establish a multiplex PCR method for simultaneous clinic detection of Mycoplasma ovipneumoniae (Mo), one of the main pathogens causing Mycoplasma pneumonia of goats and sheep(MPGS), and Corynebacterium pseudotuberculosis (CP), the pathogen of goat pseudotuberculosis, in the livestock animals infected by both pathogens at a same time. Method Two pairs of specific primers were designed based on the P80 gene of Mo and the PLD gene of CP for the PCR. After optimizing the detection system and reaction conditions, the specificity, sensitivity, and repeatability of the new method were verified for methodology validation. The results obtained using two separate PCR and the multiplex PCR on 70 clinical specimens were comparatively analyzed. Result The newly developed multiplex PCR method amplified a 700 bp fragment from Mo and a 290 bp fragment from CP, with none from other common pathogens of goats and sheep. The method had a detection limit of 1 530 pg·μL−1 on Mo and 3 500 pg·μL−1 on CP with a high repeatability. A test on 70 clinical specimens using the new method yielded a 25.71% positive rate on Mo, 18.57% on CP, and 7.14% on Mo/CP infections. Its coincidence rate with the two single PCR results was 100%. Conclusion The newly established multiplex PCR detection methodology exhibited high specificity, sensitivity, and reproducibility. It appeared to be applicable for rapid and simultaneous clinic detection on Mo and CP as well as epidemiological studies on the disease infected by the pathogens.
2020, 35(1): 51-58.
doi: 10.19303/j.issn.1008-0384.2020.01.008
Abstract:
Objective Nutritional compositions in the meat of female and male Murray cods (Maccullochella peelii peelii) at critical growth stage were determined for feed improvement of the aquaculture. Method Contents of protein, amino acids (AAs), fat, and fatty acids (FAs) in the filets were analyzed for the evaluation. Result Significant differences were found on fat and protein contents between the female and male fish fillets. Among 17 AAs, there was no significant difference between the females and males on cystine (Cys) or histidine (His) (P＞0.05). However, the females had extremely significantly higher contents of the other AAs than the males (P＜0.01). The total AA as well as the essential, non-essential, sweet, bitter, acidic, flavoring, sulfur, branch-chain, and aromatic AAs were extremely significantly higher in the females than the males (P＜0.01). The highest content of FAs in the fish filets was C18:1n9c, followed by C16:0. The C16:0 FA in meat was extremely significantly higher for the female than the male (P＜0.01), but no significant difference on C18:1n9c. The n-3/n-6 ratios were same for the female and male at 1.0. The E/T of both female and male cods surpassed the reference protein standard at 40%, and the E/NT at more than 60%, as proposed by WHO/FAO. The SRCAA was 78% for both male and female Murray cods. The filets were rich in unsaturated FAs, i.e., 57.4% for the females and 55.4% for the males. Conclusion The major nutritional components in the filets of the male and female parental Murray cods were significantly different, indicating that they ought to be fed with differently formulated forages in separate ponds. Nutritionally, the meat of either female or male Murray cods was highly recommended as a source of animal protein.
2020, 35(1): 59-66.
doi: 10.19303/j.issn.1008-0384.2020.01.009
Abstract:
Objective Diversity of bacterial and fungal endophytes on citrus plants infected with Huanglongbing (HLB) (citrus greening disease by Candidatus Liberibacter) was determined for further study on the pathology. Method The diversities of bacterial and fungal endophytes on healthy and HLB-diseased leaves of citrus plants grown in a greenhouse were compared. In addition, the diversities on the diseased leaves of different severity from a same plant were also compared. The HLB pathogen was confirmed using primer pair of OI1/OI2c. High-throughput sequencing were used to detect the 16S rRNA of bacteria and rDNA-ITS of fungi. Then the abundance and diversity analysis on the endophytes were conducted. Result For the bacterial endophytes, there were 13 phyla, 22 classes, 57 orders, 101 families, and 208 genera found, while for the fungal endophytes, 2 phyla, 12 classes, 24 orders, 43 families, and 43 genera. The relative abundance of Candidatus Liberibacter increased with severity of the disease. Methylocella and 1174-901-12 were the dominant bacteria endophytes on both healthy and diseased plants; whereas, Sphingomonas only on healthy plants. The relative abundances of Alpha cluster and Hymenobacter increased with the disease infection. Strelitziana was the dominant fungal endophyte on both healthy and diseased plants; whereas, Zasmidium and Trichomerium on diseased plants only. The relative abundance of Zymoseptoria increased with the disease infection. Conclusion The infection of HLB by Candidatus Liberibacter bacteria changed the abundance and diversity of bacterial and fungal endophytes on a citrus plant. There were also significant differences on the diversity of bacterial and fungal endophytes between the healthy and the diseased plants. Successful detection of the endophytes using high-throughput sequencing overcame the obstacles on the disease identification due to the unculturable pathogen and endophytes as well as the endophytes with low abundance. It also allowed a systematic study on the microbiology associated with HLB and the cure and prevention of the disease.
2020, 35(1): 67-73.
doi: 10.19303/j.issn.1008-0384.2020.01.010
Abstract:
Objective To identify conditions that facilitate germination of Mycogone perniciosa chlamydospores for further research on effective control of the fungal disease on edible mushrooms. Method Chlamydospores were kept at −80℃, −20℃, 0℃, 4℃, 10℃, 15℃, 20℃ or 25℃ for various durations prior to culture on media at 25℃ to determine the effects of dormancy-breaking temperatures. And effects of varied pHs, culture conditions, carbon source and nitrogen source on the chlamydospores germination were detected. Result Dormancy of the chlamydospores was broken and germination promoted under 0℃ or 4℃ for 12 h, 24 h or 48 h. pH 6, 7 and 8 were conducive to the spore germination at the rates of 8.66%, 9.79%, and 8.40%, respectively. The germination rate varied according to the medium the spores grew on. It was 0.00% on WA, 6.38% on MuEA, 8.43% on MuDA, 1.33% on PDA, and 6.23% on V8. Thus, MuDA appeared significantly better than other media tested in this study. The chlamydospores showed a germination rate of 10.77% with glucose, 9.96% with sucrose or 9.85% with sorbose in the medium, indicating a significantly advantage of the presence of a carbon source. In so far as nitrogen is concerned, the addition of proline in MuDA delivered a 12.32% germination rate, serine 13.45%, alanine 13.74%, and ornithine 11.64%, which were significantly improved over control without the addition. Conclusion By keeping M. perniciosa chlamydospores at 0℃ or 4℃ for more than 12 h could break the spore dormancy. On MuDA of pH 6, 7 or 8, the chlamydospores germinated well. With the presences of glucose, sucrose, D-lactose, sorbic sugar or D-raffinose as carbon source and L-proline, L-serline, L-alanine or L-qrnithine as nitrogen source, the spore germination was further enhanced.
2020, 35(1): 74-79.
doi: 10.19303/j.issn.1008-0384.2020.01.011
Abstract:
Objective Tests to screen and determine sensitivities of insecticides for controlling Paracoccus marginatus Williams and Granara DE Willink, an important new invasive pest on papaya plants in China, were conducted. Method The sensitivities of 2nd nymphs and female adults of P. marginatus to 15 insecticides were determined by means of leaf immersion for pesticide selection. Result Five applications of pesticides including 25 g·L−1L lambda-cyhalothrin SC, 10% chlorfenapyr SC, 4.5% beta-cypermethrin EC, 15% pyridaben EW, and 15% tolfenpyrad SC with high toxicity to P. marginatus were selected for further evaluation. Among them, the lethal medium concentration (LC50) of 25 g·L−1 lambda-cyhalothrin SC was the lowest with LC50 on the 2nd nymphs and female adults being 0.243 0 mg·L−1 and 0.782 1 mg·L−1, respectively. It was followed by 10% chlorfenapyr SC, 4.5% beta-cypermethrin EC, 15% pyridaben EW and 15% tolfenpyrad SC in that order. The 2nd nymphs were significantly more sensitive to the insecticides than the female adults. Conclusion It appeared that 25 g·L−1 lambda-cyhalothrin SC, 10% chlorfenapyr SC, 4.5% beta-cypermethrin EC, 15% pyridaben EW or 15% tolfenpyrad SC could be used to control or prevent a P. marginatus outbreak.
2020, 35(1): 80-89.
doi: 10.19303/j.issn.1008-0384.2020.01.012
Abstract:
Objective The stepwise enzymatic hydrolysis was applied to hydrolyze chicken protein for the preparation of a food flavoring ingredient with improved quality. Method An animal and a compound proteolytic enzyme were used to digest the chicken protein. Free amino acid and peptide contents in the hydrolyzed material were employed as the criteria to initially identify the critical process evaluation parameters based on a single factor Plackett-Burman design. A steepest ascent test was conducted to approximate the maximum response area and arrive at the center point in a response surface experiment. Then, according to the selected indicators for the Box-Behnken experiment, optimized process conditions were determined. Result The selected indicators for the Box-Behnken optimization included the free amino acid and polypeptide contents as well as the flavor scores on the thermal reaction products of the hydrolyzed compounds. The optimal processing conditions were found to include the addition of 0.82% enzymes on the chicken protein substrate, a solid-liquid ratio of 1 ∶ 4.40, and a reaction duration of 7.26 h to achieve an actual composite evaluation score of 88.97 in comparison to the predicted 89.41. The method was deemed reliable. Conclusion The multi-indicator response surface method effectively optimized the enzymatic proteolysis process of chicken protein materials in producing a high quality flavoring ingredient for food applications.
2020, 35(1): 90-102.
doi: 10.19303/j.issn.1008-0384.2020.01.013
Abstract:
Objective Effect of γ-polyglutamic acid on the structure of gluten network during dough-making was studied. Method Amount ofγ-polyglutamic acid added to the dough was set as variable for the experiment. Moisture and structure of the dough were observed by nuclear magnetic resonance and magnetic resonance imaging techniques. Texture, tensile properties and microstructure of the dough were measured by a texture analyzer and an environmental scanning electron microscopy. Based on these parameters, the effects of γ-polyglutamic acid additions on the gluten network at different stages of dough formation were studied. Result At 0.3% addition of γ-polyglutamic acid in the dough, the proton density M21 and M22 peaked reaching a maximum water entrapment in the protein-starch network. The proton density imaging showed a dense and uniform red portion in the dough. The texture measurements and electron microscopy imaging displayed consistent results as did the nuclear magnetic measurements. Addition of γ-polyglutamic acid prolonged the dough-making time resulting in a most stable network structure. Conclusion An addition of γ-polyglutamic acid at 0.3% in the dough with 23 min for a complete network formation would improve, but excessive additions be detrimental to, the quality of the baked bread.
2020, 35(1): 103-110.
doi: 10.19303/j.issn.1008-0384.2020.01.014
Abstract:
Objective Effects of biochar addition on the microbial community and phenolic acids in rhizosphere soil at site of continuous-cropping of tobacco at various growth stages were investigated. Method The changes on the contents of 7 phenolic acids and microbial communities in the rhizosphere soil under conventional and biochar-added fertilizations were determined by HPLC and high-throughput sequencing techniques for the analysis. Result Six out of the 7 phenolic acids were detected in the soil specimens at all tobacco growth stages. With the addition of biochar in the fertilizer, aside from cinnamic acid that showed no significant variation, the contents of p-hydroxybenzoic acid, vanillic acid, syringic acid, 4-coumaric acid, and ferulic acid in the soil were lower than those under the control treatment of conventional fertilization. Compared to control, the total phenolic acid in the soil fertilized with added biochar decreased at the resettling, vigorous growing, and harvesting tobacco stages by 1.86%, 11.36%, and 40.44%, respectively. Biochar improved the diversity and richness of rhizosphere microbial community as well. At the vigorous tobacco growing stage, the bacteria and fungi OTU abundance under biochar treatment were 1.23 times and 1.07 times of control, respectively. Compared with the conventional fertilization, the biochar addition increased the Shannon, ACE and Chao1 indices on bacteria by 12.43%, 23.0%, and 23.0%, respectively, during the vigorous growing stage; and, by 0.32%, 2.40%, and 1.25%, respectively, during the harvesting stage. On fungi, the Shannon, ACE and Chao1 indices were raised by 6.54%, 6.67%, and 7.43%, respectively, in the vigorous growing stage; and, by 57.93%, 8.60%, and 26.37%, respectively, in the harvesting stage. Conclusion It appeared that biochar significantly improved and upgraded the tobacco-growing soil by creating a healthy micro-ecological rhizosphere. The addition also alleviated some of the detrimental effects of the allelopathic autotoxicity generated from continuous cropping.
2020, 35(1): 111-116.
doi: 10.19303/j.issn.1008-0384.2020.01.015
Abstract:
Objective An analysis on balancing the ecological and economic benefits of peach orchard operations was conducted at the poverty-stricken areas in Ningde, Fujian Province to evaluate the grass-growing vs. ground-clearing practices for sustainable peach cultivation at a village scale. Method A method of analyzing emergy consumption was applied to compare the grass-growing(located at Laidun Village, Gutian County) and ground-clearing(located in Hutou Village, Fu’an County) modes practiced at peach orchards, in Fujian Province, by calculating indices of total emergy input (T), emergy self-sufficiency ratio (ESR), emergy investment ration (EIR), emergy yield ratio (EYR), environmental load ratio (ELR), emergy sustainable index (ESI), and emergy feedback rate (FYE) . Result The 2017–2018 indicators of the grass-growing and ground-clearing modes, respectively, were T at 1.16×1016 and 2.36×1016 sej·hm−2, ESR at 0.166 and 0.102; EIR at 5.030 and 8.823; EYR at 1.199 and 1.113; ELR at 0.425 and 0.410; ESI at 2.822 and 2.716; and, FYE at 0.072 and 0.026. The ground-clearing peach cultivation at Hutou employed greater inputs on both labor and organic/chemical fertilizers to resulted in a slightly lower ELR than its counterpart. In contrast, at Laidun, where grass-growing and utilization of methane were applied, led to a significantly higher FYE than at Hutou owing to a reduced demand on chemical fertilizer and emergy inputs by 57.2%. In addition, a significant 46.32% saving on emergy consumption per 10 000 RMB output of the crop was realized at Laidun. Conclusion Management of the peach orchard through grass-growing, combined with using methane liquid, has merits as an efficient and ecologically sustainable approach in the hilly villages of Ningde City.
