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Effect of peanut shell replacing peat soil on growth and quality of Anoectochilus roxburghii
ZHU Qiaolian, CHI Bingjie, TONG Chenxiao, ZHANG Hongxue, HU Kun, GUO Liming, ZHOU Biqing, MAO Yanling
, Available online  
Abstract:
  Objective  The peanut shells replaced peat soil as the cultivation substrate and the influence on the growth quality and substrate properties of the Anoectochilus sibiricum were studied to screen the optimal substrate ratio for the growth of Anoectochilus sibiricum.  Method  Through pot experiment, set five kinds of peanut shells to replace peat soil with CK (100% peat soil), N75 (75% peat soil + 25% peanut shell), N67 (67% peat soil + 33% peanut shell), N50 (50% peat soil + 50% peanut shell), N33 (33% peat soil + 67% peanut shell), explore the effects of different cultivation substrates on the physical and chemical properties of the soil and the growth and quality of the golden thread lotus.  Result  (1) The replacement of peat soil by peanut shells can improve cultivation substrate aeration, water holding capacity and fertility. Compared with the control, 33% peat soil + 67% peanut shell treatment reduces cultivation substrate bulk density by 53.84%, maximum water holding capacity and non-capillary porosity increase by 97.14% and 148.35%. Nitrogen content increased by 7.85%, available phosphorus increased by 67.76%, available potassium increased by 1830.53%, and there were significant differences; (2) 33% peat soil + 67% peanut shell as the growth condition and quality of the golden thread is the best. Plant height, root length, leaf length, and leaf width were significantly better than those of the control group; the contents of total flavonoids, polysaccharides, free amino acids, and total phenols were increased by 41.57%, 30.59%, 45.08%, and 53.16 respectively compared with the control group. %, are significantly higher than the control group.  Conclusion  Substituting peat soil with peanut shells can improve cultivation substrate fertility, promote the growth of Anoectochilus and improve the quality of Anoectochilus, and the matrix effect of 33% peat soil + 67% peanut shell is the best.
Effects of degradable bast fiber mulching film on yield and quality of netted melon and soil environment in greenhouse
TIAN Hangfei, SONG Xiaoyong, HAN Bing, CHEN Xiaofeng
, Available online  
Abstract:
  Objective  In order to further verify the degradable bast fiber film mulching cultivation effect of its application in melon crops, this study explored the effects of degradable bast fiber film mulching cultivation of netted melon growth environment and the influence of its growth and development though comparative experiments, so as to provide theoretical basis for the application of degradable bast fiber mulching film in protected cultivation.  Methods  The effect of three mulching cultivation methods on the growth and development of netted melon in solar greenhouse and the soil environment in the facility was studied using the single factor random block design method with "Luhou Tian No.1" as the test material.  Results  Compared with CK(without plastic mulch), the soil temperature of degradable bast fiber mulch (MDM) and PEBM(PE plastic mulch) could increase the inner temperature of solar greenhouse by about 3.3 ℃. MDM treatment could improve soil water content to a certain extent, and there was no significant difference with PEBM treatment. By observing the growth period of different treatments, it was found that the pollination date of the netted melon under MDM treatment and PEBM treatment was 3 days and 4 days earlier than CK, respectively. In addition, the fruit harvest date under MDM treatment and PEBM treatment was both 10 days earlier. Compared with the control group, the fruit weight of MDM treatment and PEBM treatment increased by 18.4% and 14.7%, respectively. Besides, the yield of MDM treatment was significantly higher than that of PEBM treatment, the plot yield and unit area yield were increased by about 3% compared with PEBM treatment. Compared with CK treatment, the soluble sugar content of fruits under MDM treatment and PEBM treatment increased by 10.7% and 9.9%, respectively. After netted melon harvest, the soil nutrient content was determined. The results showed that compared with CK, the soil available nitrogen in MDM treatment and PEBM treatment decreased respectively 13.0 mg·kg−1 and 9.0 mg·kg−1, soil available phosphorus decreased respectively 17.0 mg·kg−1 and 9.0 mg·kg−1, and soil available potassium decreased 33.0 mg·kg−1 and 24.0 mg·kg−1, respectively. Furthermore, total nitrogen and total potassium contents in all treatments were not significant. MDM treatment had a significant impact on the number of microorganisms in the soil, among which the number of bacteria and fungi increased the most, 101.6% and 70.66% higher than CK respectively, and the number of actinomycetes showed a slight increase, 23.24% higher than CK.  Conclusion  It appeared that the degradable bast fiber mulch cultivation was suitable for the pollution-free cultivation of netted melon.
Physicochemical Properties and Microbial Diversity of Rhizosphere Soil from Different Habitats of Pseudostellaria heterophylla in Zherong County
GAO Huifang, XU Jiayin, MENG Ting, CHEN Jinshao, QIU Junzhi, ZHANG Zhongyi, ZHANG Liaoyuan
, Available online  
Abstract:
  Objective  The purpose was to explore the potential causes of continuous cropping effect of Pesudostellaria heterophylla in different habitats.  Method  Microbial community structure and diversity of rhizosphere soil from five producing habitats in Zherong County of Fujian Province were analyzed by high-throughput sequencing technology.  Result  A total of 744331 and 1032029 effective sequences for rhizosphere fungi and rhizosphere bacteria respectively were obtained. After clustering analysis, 1314 operational taxonomic units of fungi and 10310 operational taxonomic units of bacteria were identified. The total number of mutual OTUs in five habitats was significantly more than that of exclusive OTUs in each habitat, indicating that the trend of microbial community in rhizosphere soil of Pseudostellaria heterophylla was similar in different habitats. The alpha diversity showed significant differences in the composition and abundance of microbial community in rhizosphere soil of Pseudostellaria heterophylla in different habitats. The species richness in Ⅳ, Ⅱ and Ⅰ habitats was higher than that of Ⅴ and Ⅲ habitats. Canonical correspondence analysis showed that soil chemical factors and phenolic acids in root exudates affected microbial community distribution in rhizosphere soil of Pseudostellaria heterophylla. Fusarium and other pathogenic fungi were identified in all the five habitats, and the content of pathogenic fungi was inversely proportional to the number of other microbial species in rhizosphere soil, suggesting that Fusarium affected the number of other microbial species in rhizosphere soil of Pseudostellaria heterophylla.  Conclusion  The evolution of rhizosphere microbial structure and diversity mediated by root exudate Pseudostellaria heterophylla contributed to continuous cropping effect of Pseudostellaria heterophylla to a great extent.
Introduction and Evaluation of Fresh Seed Large-grain Broad Bean Germplasm Resources in Fujian Province
LI Chengxun, LI Aiping, XU Xiaoyu, ZHENG Kaibin
, Available online  
Abstract:
  Objective  In order to introduce advanced large-grained broad bean germplasm resources to Fujian Province, the domestic advanced germplasm resources of large-grain broad bean was planted in Fujian area, and its agronomic characteristics were identified and evaluated.  Method  The study compared with 10 large-grain broad bean varieties from Qinghai and Jiangsu provinces, using the Ling-xi-yi-cun as a control which from Fujian Province. The growth period, plant height, branch number, pod length, pod width, fresh pod yield, seed length, seed width, 100-seed weight, seed rate and fresh seed yield were observed. The correlation between yield characters was studied.  Result  The results showed that the growth period, fresh pod weight, pod width, 100-seed weight of fresh seed, number of primary branches and number of pod weight were positively correlated with fresh pod weight per mu, while plant height was negatively correlated with fresh pod weight per mu. Qinghai broad bean varieties in Fujian showed short growth period, high plant type, few branches, low yield, low economic output value. Tong-can-xian varieties showed good performance and high economic value. The Tong-09-110-1’s pod length, pod width, fresh pod yield per mu, seed length, seed width, 100-seed weight, seed rate and fresh seed yield were all higher than others. Its 100-seed weight of dry seed was 200.08 g. Its seed rate per mu was 50.53%. Its fresh pod yield per mu was 1200.58 kg.  Conclusion  The Tong-09-110-1 is most suitable to be popularized and planting in Fujian area.
Generation breeding and Production performance test of Fast-growth lines of Fuqing Goat
LIU Yuan, LI Wenyang, WU Xianfeng, MAO Kunming, LIN Yunqin, HUANG Qinlou
, Available online  
Abstract:
  Objective  In order to evaluate the breeding effect of fast-growth lines of Fuqing goat,   Method  the growth performance and reproductive performance of two generations of Fuqing goat were analyzed.  Result  The results showed that after two successive generations of strict and systematic breeding, the growth and reproductive traits of the core group of Fuqing goat were improved, and the genetic progress and population uniformity of growth traits were higher than those of reproductive traits. With the increase of breeding generations, the body weight index of 2-month-old, 9-month-old and 12-month-old offspring of Fuqing goat line increased gradually. There were significant differences in body weight among three generations (P<0.05). In addition to the body weight at month age, the CV of body weight at the same measurement stage of each generation also showed a trend of gradual decrease. Compared with the zero generation, the weight of the second generation at 2 months, 9 months and 12 months increased by 7.45%, 11.74% and 10.70%, respectively. There was no significant difference in the birth weight among the progenies of the core group (P>0.05). The weaning live lamb rate and weaning lamb weight increased gradually with the increase of breeding generations. The weaning lamb weight among the three generations had significant difference (P<0.05), which were 9.42kg, 9.95kg and 10.15kg, respectively.  Conclusion  After two successive generations of systematic breeding, the growth traits of Fuqing goat fast-growth new lines have been effectively improved, and the established goal of growth traits breeding of new lines has been achieved.
Isolation, identification and histopathological observation of a case of mule duck infected with duck plague virus
ZHANG Rui, LIU Rongchang, CHEN Changfu, HUAN Yu, CHENG Longfei, FU Guanghua, SHI Shaohua, CHEN Hongmei, WANG Chunhe, FU Qiuling
, Available online  
Abstract:
  Objective  In August 2020, a flock of duck in the farm had infectious diseases in Fuzhou, Fujian province, with the incidence rate was about 65% and the mortality rate was above 90%, and there were a lot of bleeding on the skin of infected ducks. In order to identify the pathogen causing the disease and its pathogenic characteristics.  Method  The heart, liver, spleen, pancreas, kidney, skin and other tissues of the diseased ducks were collected for pathogen detection, virus isolation and identification, gene sequencing analysis and histopathological observation.  Result  Pathogen detection by PCR method, the results showed that only duck plague virus was positive in the collected tissues, and all other pathogens were negative. After that, we performed virus isolation and identification on the positive samples. As a result, duck plague virus was isolated from the visceral tissue and skin of the diseased duck, the virus was named FJ2020176. The UL2 gene of duck plague virus FJ2020176 strain was sequenced and analyzed. Our result found that the UL2 gene had 97.8%~99.9% nucleotide homology with the virulent strain CHV, CV and 2085. Compared with the vaccine strain VAC, attenuated strain 1 and attenuated strain 2, the FJ2020176 had a 528 bp nucleotide insertion in UL2 gene, which indicating that FJ2020176 was a virulent strain of duck plague virus. Histopathological observation showed that focal liver necrosis and hemorrhage were found in infected ducks; the white pulp of spleen tissue was decreased, and the lymphocytes were necrotic and exfoliated; Hemorrhage and blood stasis of kidney; hemorrhage of bursa of Fabricius and necrosis of lymph nodes; hemorrhage of skin.  Conclusion  In this study, a virulent strain of duck plague virus was isolated from the diseased ducks with massive skin bleeding, which indicated that skin bleeding was also the clinical feature of duck plague cases. These results provide new experimental data for clinical diagnosis of duck plague.
Screening of host factors interacting with Tomato Spotted Wilt Virus N protein in tobacco
MIAO Shuyue, GAO Xiaoxiao, ZHENG Limin, CHEN Jianbin, ZHAO Xingyue, CHENG Jue, CHEN Sha, ZHANG Songbai, LIU Yong
, Available online  
Abstract:
  Objective  Tomato spotted wilt virus (TSWV) is a representative member of plant negative stranded RNA viruses. The N protein of TSWV is highly expressed in the virus-infected host plants, which may regulate the infection of the virus to the host plant. In this study, the interaction between the N protein of TSWV and the host factors in the Nicotiana benthamiana was studied to explore which host proteins will be taking part in the virus infection. The results will provide a theoretical basis for further analysis of the molecular mechanism of host protein regulating infection of virus and a new idea for effective prevention and control of TSWV disease in the future.  Method  TSWV N protein was used as a bait protein, yeast two hybrid (Y2H) was used to screen the host proteins interacting with TSWV N in N. benthamiana.  Result  15 host proteins interacting with TSWV N were identified from N. benthamiana.  Conclusion  These host proteins are mainly involved in pigment biosynthesis, thylakoid membrane assembly, plant defense response and ribosome biogenesis and lipid metabolism and cell function. They may be the auxiliary proteins of translation regulation and play an important role in plant development and response to abiotic stress.
Expression of SU protein of ENTV-2FJ strain, preparation and analylsis of its polyclonal antibody
JIANG Jinxiu, ZHANG Jinpeng, LIN Yusheng, YOU Wei, HU Qilin
, Available online  
Abstract:
  Objective  In order to study the function of SU protein of enzootic nasal tumor virus of goats (ENTV-2),  Method  RT-PCR method was used to amplify the SU gene fragment from ENTV-2 FJ and cloned into pMD-19T Simple Vector; After sequencing, the cloning vector was subcloned into PET-32a (+), the recombinant plasmid was transformed into RosettagamiB (DE3) competent cells. SDS-PAGE, Western-blot and ELISA were used for identification and antigenicity analysis of recombinant proteins.  Result  The result showed that the expressed recombinant protein had 64.38 KD, and the best expression condition was induced at 37 ℃ for 4 h at a final IPTG concentration of 0.4 mmol/L. The purified ENTV-2 virus was used for SDS-PAGE, and the mice anti-SU polyclonal antibody was used as the primary antibody for Western-blot. After Western-blot analysis, the mice anti-SU polyclonal antibody could react with ENTV-2 antigen specifically. It is proved that the expressed SU recombinant protein has better antigenicity.  Conclusion  It lays the foundation for the preparation of ENTV-2 specific monoclonal antibody and the establishment of ENTV-2 specific serological method.
The study on tea plant cultivars resistance to green leafhopper based on PPIN
WANG Fen, PEI Huimin, WEN Di, CHEN Zhi, LI Jing
, Available online  
Abstract:
  Objective  Protein-protein interaction network(PPIN) is very important in life science. It is of great significance to analyze the molecular mechanism of insect resistance of tea.  Method  RNA-seq was performed to analyze NiaoWang infected by green leafhopper or not. PPIN of differential expression genes among 0h, 12h, and 24h were constructed based on Interolog.  Result  The results showed that there were 212 PPIs between 0 h and 12 h, 3 551 PPIs between 0 h and 24 h, 3 605 PPIs in the merged network. And the topological properties of the PPINs were analyzed, in which the degree of the proteins followed power-law distribution. The function of proteins can be predicted through PPIN. PPIs of DGEs were mainly taking part in plant pathogen interactions, plant hormone signal transduction, DNA base excision repair, nucleotide excision repair, mismatch repair, linolenic acid and alpha-linoleic acid metabolism and other biological processes. In addition, leaves were infection of 24 h need to start more PPI than 12h to resisance to green leafhopper.  Conclusion  The result provide reference for the molecular mechanism of tea resistance to green leafhopper and offer basis of the control of diseases and improving breeding.
Cloning and sequence analysis of sucrose synthase gene in alpine potato in Huaiyushan(Solanum tuberosum L. cv. Huaiyushan)
HONG Senrong, DENG Yuqing, WU Hongting, CHEN Ting, GUO Shuzhen, YANG Yuxuan, CAI Hong, CHEN Ronghua
, Available online  
Abstract:
  Objective  To analyze the nucleic acid sequences information of sucrose synthase (SuSy), the key enzyme in sucrose metabolism pathway, and predict the protein structure and further reveal the biological function of the enzyme.  Method  The core fragment (PGSC0003DMG400002895, SuSy 4) of sucrose synthase gene was screened from the transcriptome database of plantlets of alpine potato of Huaiyushan. Cloning of sucrose synthetase gene of potato of huaiyushan mountain by RT-PCR and the sequence was analyzed by bioinformatics method.  Result  The total length of cDNA of sucrose synthetase gene in alpine potato of Huaiyushan was 2418 bp, and the content of G+C was 45.08%; The protein of sucrose synthetase gene in alpine potato of Huaiyushan was composed of 805 amino acids with molecular weight of 92471.33 Da and isoelectric point of 5.87, which was hydrophilic protein; The secondary structure of sucrose synthetase gene in alpine potato of Huaiyushan was composed of alpha helix (45.84%), β-Extended strand (15.16%), random coil (39.01%). C-terminal and N-terminal contain β-lamellae and α-helix, while irregular curl, extension chain, β-lamellae and α-helix are scattered in the whole protein. The tertiary structure of sucrose synthetase gene in alpine potato of Huaiyushan is tetramer. The sucrose synthase in alpine potato of Huaiyushan mainly exist in cytoplasm, mitochondria and chloroplast. The sucrose synthase gene of alpine potato of Huaiyushan was closely related to Solanum lycopersicumSolanum pennelliiSolanum chilenseSolanum tuberosumCapsicum annuumand Capsicum baccatum in evolution, especially has the highest phylogenetic relationship with Solanum tuberosum.  Conclusion  The sucrose synthase gene in alpine potato of Huaiyushan has the structural characteristics of typical sucrose synthases. The amino acid sequence and nucleic acid sequence are highly similar to the homologous species, highly conserved in evolution, which is of great significance to further reveal the biological function of the enzyme.
Effects of Rice Straw Biochar on Dynamic Changes of Water Soluble Cd in Three Different Soils
ZHANG Jiakang, ZHUAN Yalin, ZHANG Liwen, LIN Guiquan, LIN Yongcong, LI Honghong
, Available online  
Abstract:
  Objective   To investigate the immobilization effect of biochar on cadmium (Cd) in different kinds of soils under flooding conditions.   Method   A flooding incubation experiment was conducted by the addition of biochar to yellow soil, paddy soil derived from quaternary red clay, and brown soil which were contaminated by Cd.   Result   The results showed that biochar reduced soil solution pH and soil redox potential, but increased soil conductivity at the first day of incubation. With the incubation time increased, soil redox potential of three kinds of soils were reduced sharply, but biochar inhibit the reduction reaction. At the first day of incubation, the Cd content in three soil solutions was in the order: yellow soil (272.5 mg·L−1)>paddy soil (23.48 mg·L−1)>brown soil(1.44 mg·L−1), and biochar reduced the Cd content in three soil solutions by 31.66%, 75.04% and 66.67%, respectively. With flooding time increasing, Cd content in soil solution of the control treatment decreased gradually, while that was decreased slightly under biochar treatment. After 30 days of flooding, biochar significantly reduced the content of CaCl2-Cd in yellow soil, paddy soil and brown soil was reduced by 17.3%, 56.3% and 12.4%.   Conclusion   Biochar reduced the water-soluble Cd content of the soils, but the difference of soil water-soluble Cd content between control and biochar treatment decreased with the incubation time increasing. The immobilization effect of biochar on soil available Cd in paddy soil was better than that of yellow soil and brown soil, under flooding conditions.
Genetic diversity analysis of phenotypic traits of the germplasm resources of Blumea balsamifera (L.) DC.
XIAO Yongfeng, HUANG Mei, YU Fulai, CHEN Zhenxia, LIAO Li, PANG Yuxin
, Available online  
Abstract:
  Objective  The analysis of Blumea balsamifera (L.)DC. germplasm diversity of phenotypic traits can provide a scientific basis for breeding.  Method  Based on the 159 samples of B. balsamifera germplasm resources, 9 quantitative traits and 13 qualitative traits are determined, and the diversity was analyzed by adopting genetic diversity analysis, correlation analysis, principal component analysis and cluster analysis.  Result  As a result, the phenotypic traits of 159 materia showed a rich genetic diversity. Among the quantitative traits, the highest genetic diversity index was plant height (2.072), and the highest coefficient of variation was the number of flower branches (32.76%). While in the qualitative traits, the highest genetic diversity index was leaf shape (1.201). There exists a complex relationship in the traits. In quantitative traits, there was a most significant positive correlation between the crown breadth and the plant height, leaf width, petiole length, length of flower branch and opening angle of flower branch (P<0.01). In the qualitative traits, the color intensity of anthocyanin in leaf vein and the color intensity of anthocyanin in main stem, the color intensity of anthocyanin in leaf edge and the color intensity of anthocyanin in petiole had a most significant positive correlation. The cumulative contribution rate of the first 8 principal components reached 64.32%, and the factors were successively named as yield factor, color-intensity factor, leaf smoothness factor, leaf edge factor, leaf shape factor, leaf greenness factor and flower-branch angle factor according to the load amount of each trait. Based on phenotypic traits to the sum of squares deviations method in genetic distance of 10 selected materia can be divided into three groups. Group Ⅰ has 39 materia, accounting for 24.53% of the total. Group Ⅱ has 38 materia, accounting for 23.90% of the total. Group Ⅲ has 82 materia, accounting for 51.57% of the total.  Conclusion  The phenotypic traits of 159 B. balsamifera germplasm resources were rich in genetic diversity, and the leaf width could be used as the guiding target trait for breeding high-yielding B.balsamifera germplasm in the future.
Suppressed Expression of SlSIP1L12 Gene Stimulates Seeds Germination in Tomato
CUI Baolu, CHEN Guoping
, Available online  
Abstract:
  Objective  The role of SlSIP1L12, one SIP1 subfamily gene of trihelix transcription factors, was identified in tomato.  Method  Expression pattern and responses of SlSIP1L12 to phytohormone and abiotic stress were tested by RT-PCR. The RNAi technology was used to suppress SlSIP1L12 expressions to discovering its function. The ELISA was applied to detect endogenous ABA contents of seeds.  Result  (1) The length of SlSIP1L12 coding sequence was 1125 bp in AC++, which encoded 374 amino acids. Phylogenetic analysis suggested that SIP1 genes have obviously evolved in AC++. (2) Expression pattern displayed that SlSIP1L12 was mainly expressed in stem and mature leaves, and secondly in flowers and fruits. (3) SlSIP1L12 expressions could be induced by ABA and dehydration, which implied the correlation between SlSIP1L12 and ABA. (4) The germination speed of transgenic seeds was faster than control along with longer radicles. (5) ABA contents were remarkably decreased in transgenic seeds after 7 d germination.  Conclusion  SlSIP1L12 negatively regulated the germination speed of tomato seeds, which was related to decreased ABA responses.
Physiological and biochemical analysis of liquid spawn of Agaricus bisporus W192 during shaking flask culture
DAI Jianqing
, Available online  
Abstract:
  Objective  The regularity of mycelium growth in flask culture of liquid spawn of Agaricus bisporus W192 was studied in order to provide reference for the determination of liquid spawn vitality  Method  The variation of physiological and biochemical indexes in the biomass of mycelia of A. bisporus, the number and diameter of mycelium pellets, the pH value of fermentation broth, the content of reducing sugar and amino nitrogen, the activity of extracellular enzymes activity of carboxymethyl cellulase, amylase, acid protease and laccase were determined by shaking flask culture.  Result  On the 8 d of culture, the mycelium biomass of A. bisporus was the largest, 9.7 mg· ml-1.The largest number of mycelium pellets was 880 CFU·mL-1. The average diameter of mycelium was the largest, 0.809 mm. The contents of reducing sugar and amino nitrogen in the culture medium were the highest, 5.228 and 0.079 mg· ml-1, respectively. Carboxymethyl cellulase, amylase and laccase had the highest activity (0.69 U, 2.11 U, 15.02 U, respectively). On the 10th day of shaking flask culture, the activity of acid protease was the highest, and the enzyme activity was 2.93 U. The maximum number of mycelium pellets was 115 CFU·mL-1 and the maximum mycelium biomass was 7.56 mg·mL-1 when the liquid spawn cultured in shaking flask for 8 days were transferred to the liquid medium for the simulated fermentation tank liquid culture.  Conclusion  The liquid spawn activity of A. bisporus has certain correlation with the above indexes. Combined with the verification of the activity of liquid spawn, it is judged that the liquid spawn in the 8 d has the highest activity, which can be used as liquid spawn for fermenter propagation culture.
Development of Visual Loop-Mediated Isothermal Amplification (LAMP) D etection of Salmonella in Vegetable Cultivated Soil
LYU Xin, LIU Lanying, CHEN Lihua, LI Yue-ren
, Available online  
Abstract:
  Objective  To develop a visual loop mediated isothermal amplification (LAMP) for rapid detection of Salmonella in vegetable cultivation soil, and provide technical support for the prevention and control of Salmonella in vegetable cultivation soil.  Method  LAMP primers were designed based on the sequences of invasion protein A (invA) gene of Salmonella, and the LAMP reaction system and reaction conditions were optimized. LAMP detection for Salmonella was performed by specificity, sensitivity test and artificially inoculated test. Moreover, the LAMP method was verified by comparing with the national standard detection method through Salmonella detection in vegetable cultivation soil.  Result  The results of specificity test showed that the LAMP reaction of 6 non-Salmonella strains were negative except for Salmonella, which indicated that the LAMP method had good specificity, the sensitivity test showed that the minimum detection limit of the LAMP method was 7 CFU/25 μL, and the artificially inoculated test showed that the detection sensitivity for Salmonella was 4×102 CFU·g−1. Compared with the national standard method, the LAMP method had the same accuracy for Salmonella detection in vegetable cultivation soil.  Conclusion  This study developed a rapid, reliable and sensitive constant temperature amplification system, which can be applied to the rapid detection of Salmonella in vegetable cultivated soil.
Isolation and identification of a novel goose astrovirus FJ-NP
LIN Yusheng, JIANG Jinxiu, ZHANG Jing-peng, YOU Wei, HU Qi-lin
, Available online  
Abstract:
  Objective  In order to identify the pathogen of an infectious disease characterized by gout with the incidence rate of gout was as high as 40% and the mortality rate was 80% among young geese aged 5 to 20 days, broke out in a goose farm in Nanping city, Fujian province in august 2019.  Method  In this study, three samples collected in this farm were tested in laboratory and identified by pathogen isolation.  Result  The PCR detection results showed that novel goose astrovirus was detected all the three samples. Bacteria isolation result showed that no isolated bacterial infection. Vaccination age of 11, goose embryo death did not appear, but the body skin appears dotted hemorrhage, kidneys, lungs all have haemorrhage, the allantoic fluid and tissue samples were collected for PCR detection results show that the only positive for goose stellate, no other goose common virus was detected, named FJ-NP; The genetic evolution analysis of partial ORF2 genes of the isolation strain showed that the isolation strain had a closer relationship with the Anhui (GD AHAU2, AHAU3 and AHAU5), Heilongjiang and Henan (AstV-HN02-Goose-1119-18、AstV-AH02-Goose-0715-18、AstV-HB02-Goose-0310-19、AstV-HN03-Goose-0402-19) strains and had a far relationship with to Hubei, Beijing and Fujian strains.  Conclusion  In this study, a novel strain of goose astrovirus was successfully isolated, and the genetic evolution analysis of partial ORF2 genes was carried out, which provided materials for further studies on the novel goose astrovirus in in our province.
Prokaryotic expression, protein purification of Tobacco mosaic virus P54 gene
LI Cong, TIAN Peijie, ZHANG Yu, ZHANG De yong, LIU Yong, ZHANG Songbai
, Available online  
Abstract:
  Objective  Tobacco mosaic virus (TMV), a typical member belongs to genus Tobamovirus of Vigaviridae, which could infect over four hundred species of plants of 36 families, and result in 20~30 % of yield loss, or huge yield loss in case of serious damage, the result of pathogenicity and its molecular mechanisms should uncover the potential threat against plants by TMV. The genome of TMV was plus single strand RNA including 6400 bp nucleotides, which encodes one structure protein and two unstructure proteins. P 54 gene was inserted in P183 gene, which functions involving in viral replication. The functions and related molecular mechanisms were still blurried. To obtain purified samples of P54 protein, and to lay the foundation for the functional study of P54 protein.  Methods  The sequence of P54 gene (1425 bp) was amplified by RT-PCR using cDNA of Nicotiana tabacum var. Samsun NN infected with TMV, cloned into prokaryotic expressing plasmid pEASY-Blunt E1, and expressed in E coli BL21 (DE3) by IPTG induction. The expression products were retrieved and purified by Ni-NTA chromatography, and identified by Western-blotting.  Results  The expressed products of recombinant P54 protein were insoluble, and retrieved and purified P54 protein was about 60 kDa, and verified by Western-blotting.  Conclusions  The results of this study that the TMV P54 protein were successfully expressed and would be contributed to characterize the functions of P54 protein.
Rapid identification of Bactrocera tau (Diptera: Tephritidae) by using species-specific PCR technique
HUANG Zhen, Hou YouMing, GUO QiongXia, CHEN ShaoPing
, Available online  
Abstract:
  Objective  Rapid identification of the Bactrocera tau (Walker) to prevent the introduction and spread of the pest.  method  This paper based on ss-pcr technology, 20 kinds of common fruit flies with similar morphology were selected, and B. tau (Walker)were used as positive control. Genomic DNA templates were extracted, PCR amplification was conducted, and their products were tested by electrophoresis.Choose COⅠ mt DNA sequence, check the homologous series, designed a fast and accurate identification of B.tau species specific primers NF404 and NR610.  results  Only target species can amplify the clear and about 207 bp single band the rest no band occurred.  conclusion  Verify the experiment repeatedly this method specificity is strong, appraisal is accurate, and can quickly identify the fruit fly each state even residue, It is a good method of specific identification that can be applied to the rapid identification of Entry and exit inspection and quarantine and monitoring.
Preparation and Regeneration Method of Didymella bryoniae Protoplast in Melon
MAO Jiancai, WANG Haojie, LI Junhua, ZHAI Wenqiang
, Available online  
Abstract:
  Objective  For better understanding the pathogenicity of Didymella bryoniae in melon, a highly efficient system of preparing and regenerating protoplast should be established.  Method  In this study, two lyases, 20 g/L Driselase and 8 g/L Lysing enzymes were used to prepare protoplasts.The singlefactor experiment was used to analyze the effects of mycelium age, enzymatic digesting time and temperature, rotate speed, type and concentration of osmotic stabilizer and pH on the release of protoplasts.The conditions of regeneration medium were also optimized.  Result  The results showed that the yield of protoplast was the highest with 9.65×107 cells/mL under the optimal conditions: collecting the mycelia cultured for 36 h, using 0.7 mol/L NaCl as osmotic stabilizer, then culture at 140 r·min−1, and lysing for 4 h at 30 ℃, the pH of the enzymatic solution was set to 7.0.The protoplast regeneration rate was up to 22.53% in SR culture medium that the agar concentration keep 0.5%; The protoplast regeneration rate decreased rapidly within 10 h after storage, and gradually decreased gently after 10 h.  Conclusion  The efficient protoplasmic preparation and regeneration system was determined, which laid a foundation for the establishment and research of genetic transformation system of this strain.
Microalgae for Effective Nitrogen and Phosphorus Removal from Aquaculture Effluence
JIA Wei, NIE Yilei, CHEN Hong, LUO Lijin, LE Zhanxian, ZHUANG Hong, ZHENG Junrong
, Available online  
Abstract:
  Objective   Microalgae capable of effectively removing nitrogen and phosphorus from wastewater were screened for potential applications on antipollution or effluence treatment at aquaculture farms.   Method   Three selected microalgae, Chlorella sp. JY-1, Chlorella sp. SY-4, and Desmodesmus sp. SH-1, were evaluated for their capability in removing nitrogen and phosphorus from the wastewater at a shrimp aquaculture farm.   Result  After 5d of cultivation, the microalgae grew to a cell density in the 7±1‰ salinity medium at 1.56×107·ml−1 on JY-1, 1.47×107·ml−1 on SY-4, and 6.62×106·ml−1 on SH-1. The removal rate on total nitrogen was 50.36% by JY-1, 41.51% by SY-4, and 49.74% by SH-1; that on ammonia nitrogen, 96.29% by JY-1, 84.92% by SY-4, and 96.65% by SH-1; that on nitrate nitrogen, 15.84% by JY-1, 3.69% by SY-4, and 12.56% by SH-1; and, that on total phosphorus, 93.51% by JY-1, 82.38% by SY-4, and 94.25% by SH-1; but not significant on nitrite nitrogen by any of them. The microalgae appeared to grow normally in a culture medium of 5%, 10%, 20% or 30% salinity. Among them, Chlorella sp. JY-1 performed the best on growth as well as nitrogen and phosphorus removal in the test wastewater.   Conclusion   Chlorella sp. JY-1 appeared to offer a promising potential for application of purifying aquaculture effluence.
Growth and Nutrient Content of Dendrobium officinale as Affected by Epulorhiza Symbiosis
WANG Weiying, ZOU Hui, DAI Yimin, LIN Jiangbo
, Available online  
Abstract:
  Objective  Effects of the symbiosis between Epulorhiza sp. and Dendrobium officinale in a liquid culture medium on the growth and nutrient content of the plant were analyzed.  Method   In a potted experimentation, the sterile D. officinale seedlings were irrigated with a liquid medium as control or one containing Epulorhiza sp. for the treatment. Agronomic characteristics and nutrient contents of the stems and leaves from one- and 2-year-old D. officinale plantlets were monitored.   Result  Showing dark green leaves and robust stems, the treatment plantlets grew more vigorously than control. In the presence of Epulorhiza sp., on average the girths of the productive stems of one- and 2-year-old plantlets significantly increased by 65.67% and 74.25%, respectively, and the single stem weight by 55.29% and 51.45%, respectively (P < 0.05). The increases meant improved crop yield. Meanwhile, the contents of polysaccharides, dendrobine, crude protein, and amino acids increased as well. For the one-year-old plantlets, the crude polysaccharides rose significantly by 30.39%, and the crude protein 18.7% (P < 0.05). For both one- and 2-year-old plantlets, the total amino acids significantly increased by 27% and 30.25%, respectively (P < 0.05), while the crude fiber and ash significantly decreased by 17.76% and 36.36%, respectively (P < 0.05).   Conclusion  Both crop yield and nutrient content of D. officinale could be significantly improved by the presence of Epulorhiza sp. in medium. It suggested the potential benefits of organic fungi-containing manure utilization, artificial greenhouse operation, and harvest time management for the cultivation of D. officinale.
Guangxi Subtropical Crops Research Institute, Guangxi Academy of Agricultural Sciences, Nanning, Guangxi 530001, China
LI Jidong, YANG Xiangyan, CAI Yuanbao, LI Mu, ZENG Liming, HUANG Sijie, WU Fumin, LIN Yuhong, ZHENG Wenwu, HUANG Jinyuan
, Available online  
Abstract:
  Objective  Two new protein phosphatase genes, MiSTPP1 and MiSTPP4, from macadamia nuts (Macadamia integrifolia) were cloned for structure and function analyses by bioinformatics.   Method  The genes were cloned from M. integrifolia using transcriptome sequencing and RT-PCR technique. Homology, phylogenetic evolution, physicochemical properties, phosphorylation sites, subcellular localization, transmembrane domains, and signal peptides of the genes were analyzed.   Result  The cloned MiSTPP1 and MiSTPP4 from M. integrifolia were assigned with the GenBank accession numbers MT374548 and MT374551, respectively. The amino acid sequences of the genes had a similarity to other plant PP1 proteins containing the same typical structural domain MPP_PP1_PPKL. The evolutionary tree analysis showed that they closely related to PP1 family proteins. The basic physicochemical properties of MiSTPP1 indicated it be an unstable hydrophilic protein, and those of MiSTPP4 a stable hydrophilic protein. Phosphorylation sites on the genes were principally serine and threonine. The predicted subcellular localization, transmembrane domain, and signal peptide suggested that MiSTPP1 and MiSTPP4, as non-secretory and non-transmembrane proteins, were most likely located in the cytoplasm. The secondary and tertiary structures of the genes contained mainly α-helixes and random coils.   Conclusion  MiSTPP1 and MiSTPP4 from macadamia nuts belonged to the protein phosphatase PP1 gene family and might play a role in response to stress and signal transduction as well as other physiological and biochemical processes of the plant.
Codon optimization and expression of bovine pregnancy associated glycoprotein-16 gene
LIU Changbin, LU Chunxia, LU Shouliang, NI Jianhong
, Available online  
Abstract:
  Objective   The bPAG16 gene was optimized and synthesized by bioinformatics techniques. The objective of this study to construct the recombinant expression vector of proEM-bPAG16 and expressed it in HEK293 cells, which provided the technical support for the research of pregnancy diagnosis products.   Methods   The bPAG16 gene was optimized and synthesized by bioinformatics techniques. Then, the codon-optimized bPAG16 gene was connected with proEM vector by T4 DNA Ligase. The proEM-bPAG16 recombinant vector was transfected into HEK293 cells. The recombinant bPAG16 fusion protein was purified by Ni2+ affinity chromatography and evaluated by SDS-PAGE and western blotting analysis.   Results   After codon optimization, the codon adaptation index (CAI) of the bPAG16 gene raised from 0.77 to 0.96 and its GC% content changed from 48% to 58%. After enzymatic digestion, The ~ 1179 bp and ~ 4369 bp fragments were obtained from proEM-bPAG16 expression vector, which was consistent with the expected value. Sequence analysis showed that the nucleotide sequence of the inserted bPAG16 gene in recombinant plasmid was consistent with the optimized bPAG16 gene, the amino acid was not mutated. SDS-PAGE and western blotting showed that the recombinant bPAG16 fusion protein (48 kDa) was correctly expressed in HEK293 cells. Recombinant bPAG16 purity was above 90%.   Conclusion   This laid a foundation for the further study of dairy cattle early pregnancy diagnosis.
Advances in genetic and breeding of functional components in colored rice
ZHENG Feiyan, ZHENG jianhua, WANG Hongfei, ZHU Yongsheng, YOU Qingru, ZHOU Peng, CHEN Chunxia, TU Shihang, DONG Ruixia, ZHENG Jiatuan, Huang Tingxu
, Available online  
Abstract:
With different anthocyanins content in the seed coat, the brown rice present different colors such as black, purple, red, green, yellow and other colors, which was called colored rice. Colored rice has more abundant nutrition and health value than common rice (white rice). Colored rice contains various beneficial substances which human needs, such as amino acids, functional lipids, dietary fiber, vitamins, minerals, anthocyanins, phenolic compounds, γ-oryzanol, and so on. It has important nutritional and medicinal value, which enjoys high popularity among consumers and well market’s attention. It has great market potential. The germplasm resource distribution, evaluation and utilization, functional components and their characteristics, genetics research and breeding application of the colored rice were reviewed in this paper. The problems restricting the development of colored rice industry were analyzed in this paper, at present, colored rice is lagging behind the common white rice varieties, new functional foods and medicines of colored rice are yet to be developed. In order to promote the innovative development of China's colored rice industry, it is necessary for the market and industrial development to carry out comprehensive research on colored rice systematically and effectively, the breeding efficiency of colored rice varieties can be greatly improved by strengthening the assistant breeding with biological means .
Identification of pathogens Causing diseases on Chinese yam planted on the ridge of the northern area in Hubei Province
YANG Xiaolin, ZHANG You hong, WANG Zuoqian, CHANG Xiangqian, LV Liang, ZHANG Shu, YANG Xinsun
, Available online  
Abstract:
  Object  The study was to explore the pathogens causing diseases on yam on the ridge of the northern area in Hubei province and their classification status by identification of pathogens causing diseases, so as to provide the basis for further study of pathogens’ occurrence and draft of effective prevention and control settlement.  Method  Disease samples of the aboveground parts of yam were collected and pathogens of yam disease were isolated and identified through colonial morphology, microstructure, blasting sequence of ITS of pathogens and determination of pathogenicity by means of tissue separation of phytopathology and ITS-PCR technique.  Result  The result showed that the pathogenic fungi include Alternaria/Alternaria alternate, Colletotrichum /Colletotrichum fructicola and Colletotrichumn gloeosporiaides, and Lasiodiplodia/Lasiodiplodia pseudotheobromae.  Conclusion  The above results show that several diseases of yam on the ridge of the northern area in Hubei province often occur and composition of the pathogens is complex. Diseases include Chinese yam anthracnose and other diseases, which all can cause Chinese yam tissues to wither to death.
Study on Bacteriostatic Characteristics of Lactobacillus plantarum LV02 and Optimization of Fermentation Medium
YANG Yue, HUA Wanyue, CHEN Zhidi, ZHANG Yao, YI Xinxin, GAO Xiuzhi
, Available online  
Abstract:
  Objective   To study the bacteriostatic characteristics of Lactobacillus plantarum LV02 and optimize its fermentation medium.   Method   Based on the single-factor experiment, the PB experiment and the steepest ascent experiment were used to determine the step length and direction of the influencing factors, and the CCD experiment center point is determined. A CCD design experiment was used to study the effect of fermentation medium on the bacteriostasis of Lactobacillus plantarum LV02 and OD600, and determined the optimal level of each influencing factor. The Oxford Cup method was used to study the bacteriostatic characteristics of LV02 to determine the stability of LV02 in the application environment.   Result   The optimize fermentation medium formula of LV02 was: glucose 34.07 g·L−1, yeast extract 18.12 g·L−1, dipotassium hydrogen phosphate 2 g·L−1, manganese sulfate 0.16 g·L−1, sodium acetate 5 g·L−1, magnesium sulfate 0.20 g·L−1, ammonium citrate 1 g·L−1, tween 80 1 ml·L−1, carrot juice 50 ml·L−1, and distilled water 1 L. After optimization, the diameter of the inhibition zone of LV02 against Escherichia coli YS increased by nearly 26% compared with that before optimization, and OD600 increased by 12%. The saturation concentration of ammonium sulfate required for crude extraction of bacteriocin of Lactobacillus plantarum LV02 was determined to be 80%, which proved that Lactobacillus plantarum LV02 was stable to heat (100 ℃, 120 min), acid-base stability (pH 3.0~7.5) and antibacterial.   Conclusion   Optimize the fermentation medium of Lactobacillus plantarum LV02, studied its bacteriostatic characteristics, and provided technical reference for the development of antibacterial and fresh-keeping products.
Response of wheat Aikang 58 seedlings to Na2SO4 stress and determination of threshold
LIU Duo, BAI Shuang, QI Xuebin, NING Dongfeng, LIANG Zhijie, GUO Wei, LI Ping
, Available online  
Abstract:
  Objective   Bainong Aikang-58 is the main cultivar of wheat in China. The response mechanism of Bainong Aikang-58 seedling to Na2SO4 stress was discussed and its tolerance threshold to Na2SO4 was determined so as to provide basic data support for further research and scientific basis and theoretical guidance for agricultural production practice.   Method   In this study, the mechanism of Na2SO4 stress at different concentrations (0, 20, 40, 60, 80 and 100 mmol·L−1) on the growth and physiology of wheat seedlings was studied through hydroponic experiments.   Result   It was found that with the increase of Na2SO4 concentration, the plant height, fresh weight and biomass of wheat seedlings all decreased, The activity of each antioxidant enzyme in wheat seedlings would continuously increase with the increase of Na2SO4 concentration, and would decrease after exceeding a certain threshold. The permeability of MDA content in synplasmic membrane would continuously increase with the increase of Na2SO4 concentration, The maximum values were 3.2 times and 4.94 times of the control, respectively. the chlorophyll content of wheat seedling leaves decreased with the increase of Na2SO4 concentration. The soluble sugar content in wheat increased first and then decreased with the increase of Na2SO4 concentration. The soluble sugar concentration was the highest at 80 mmol·L−1, and the proline content kept increasing with the increase of Na2SO4 concentration, The maximum value is 8.51 times that of the control.   Conclusion   According to the experimental results, the Na2SO4 stress tolerance threshold of Aikang 58 was defined as 80~100 mmol·L−1.
Cloning and Post-harvest Physiological Deterioration Expression of MePYL12 of Cassava
GUO Jing, ZHANG Yuxiang, HUANG Zhiyi, WU Chunlai, Yan Yan, ZENG Jian, HU Wei
, Available online  
Abstract:
  Objective  Functions of PYL gene in the post-harvest physiological deterioration (PPD) and under abiotic stresses of cassava (Manihot esculenta Crantz) were studied for further research on the key component of the plant abscisic acid (ABA) signaling pathway, PYR/PYL/RCARs family.   Method  The PYL gene, MePYL12 was cloned from cassava SC 124 by RT-PCR. Bioinformatics was used to analyze the physicochemical properties, conserved domain, genetic evolutionary relationship, protein structure prediction, promoter elements as well as the expressions of MePYL12 in PPD and under abiotic stresses.  Result  (1) The full-length cDNA of MePYL12 was 567 bp encoded a polypeptide of 188 amino acids with a predicted relative molecular mass of 20.8kD and an isoelectric point of 5.55. The predicted tertiary structure contained a helix. Its multiple protein sequence alignment showed high similarities with the PYL proteins in Hevea brasiliensis (94.68%) and Ricinus communis (86.77 %). MePYL12 consisted of the conserved motifs of the PYL family, such as the ABA binding region "Latch" and "Gate" suggesting it to be a genuine member from the family and highly conserved. (2) The expressions of MePYL12 in 10 different types of cassava tissue/organ were high in the root apical meristem, shoot apical meristem, and leaf. (3) The promoter element analysis showed that the gene contained the light-responsive, drought-induced MBS, and ABA responsive ABRE motifs. (4) The expression of MePYL12 was significantly upregulated by ABA treatment and drought stress as well as during PPD that peaked in 6h followed by downregulation.   Conclusion  MePYL12 might participate in the PPD process improving the ability of cassava plants in dealing with abiotic stresses. Further study on the functions of MePYL12 in cassava is in order.
BSA-Seq Identification of Blast-resistance Genes in Gufeng B Rice
CHEN Ziqiang, CHEN Songbiao, GUO Xinrui, YAN Jingwan, TIAN Dagang, LI Gang, WANG Feng
, Available online  
Abstract:
  Objective   To identify and map the blast-resistance gene(s) in Gufeng B, a rice cultivar that confers strong, broad-spectrum, durable resistance against various blast strains.   Method   F1 and F2 population of the cross between Gufeng B and Nipponbare were inoculated with Magnaporthe grisea 501-3 to analyze the inheritance mechanism of the blast resistance in rice. From F2, cultivars extremely resistant or sensitive to the blast were selected to establish contrasting DNA pools. Subsequently, the associated loci were mapped by means of the bulked segregation analysis (BSA-Seq).   Result  Gufeng B exhibited high resistance to all tested strains including KJ201, RB22, CHNOS, RB6, 2Y838-1, 501-3, and IR16-1. It suggested that the cultivar carried broad-spectrum, highly resistant genes toward the disease. And, since the F1 progenies showed apparent resistance against 501-3 and IR16-1, but F2 failed to display a 3 1 ratio on the resistance vs. susceptibility separation, the disease resistance was presumably controlled by multiple loci in Gufeng B. The re-sequencing of entire genomes on the DNA pools of F2 and the parental Gufeng B and Nipponbare identified 1 756 964 SNPs. The calculated △SNP-index indicated two candidate loci related to the blast resistance to be located at Chr.6: 10 082-11 397 Kb and Chr.11: 120-266 Kb. Of the two, Chromosome 6 had a region containing 4 006 SNPs and 623 InDels markers that corresponded to the Pi2/9 disease resistance locus, while Chromosome 11 had one consisting of 752 SNPs and 195 InDels.   Conclusion   The resistance of Gufeng B to 501-3 blast strain might be jointly regulated by the responsible genes in Chromosome 6 and Chromosome 11. The information would be of value in refined mapping and cloning of the resistance genes in Gufeng B as well as in selecting markers for the related studies.
Application of Fe3O4 Nano-particles in the Pesticide Residues Detection for Tea after Method-optimization
LI Jie, ZHANG Feng, HUANG Jingjing, DING Liping, YANG Fang, YAO Qinghua
, Available online  
Abstract:
  Objective  The aim of this study were to develop a new analytical method for detecting 7 pesticide residues in the tea, which was help to improve the efficiency and reduce the cost. Fe3O4 Nano-particles was used to remove the pigments in tea extracts, then residues of carbendazim, pyrimethanil, tricyclazol, imidachloprid, acetamiprid, methomyl, and thiamethoxam were detected by liquid chromatography-tandem mass spectrometry. The validity of method were evaluated.  Result  The recoveries of 7 pesticide residues in Fe3O4 Nano-particles group was better than that in graphitizing of carbon black (GCB). The optimism dosage of Fe3O4 Nano-particles was 300 mg. The purified solution of tea extracts were separated with a Phenomenex Luna C8 (150 mm × 2.0 mm × 3.0 µm) by using (0.1 % formic acid and 5 mmol/L ammonium acetate) - acetonitrile as mobile phase, then analyzed by tandem mass spectrometry under multiple reaction monitoring (MRM) mode with external standard method. The linearity of method was good for 7 pesticides during the mass concentrations of 0 ~ 50 µg/L, and the correlation coefficients more than 0.995. The average recoveries of pesticides were in the range of 71.6 %~107.7 % at three spiked levels (5 ng/g, 10 ng/g, and 50 ng/g) with relative standard deviations of 3.95 %~13.62 %. The limits of detection of pesticides were from 0.15 to 0.6 µg/kg. The limits of quantitation (LOQ) of pesticides were from 0.5 to 2 µg/kg.  Conclusion  The proposed method was stable and had good repeatability, which is helpful to improve the recoveries of pesticide residues in tea with reducing the detection cost.
Transcriptome Analysis on Tubers of Two Huaiyushan Cultivated Varieties of Tetrastigma hemsleyanum
HONG Senrong, HUANG Dandan, HUANG Shihui, HUANG Xiameng, JIANG Ye, LI Wanping, CAI Hong, CHEN Ronghua
, Available online  
Abstract:
  Objective  Transcriptomes of differentially expressed genes related to flavonoids synthesis in tubers of two Huaiyushan cultivated varieties of Tetrastigma hemsleyanum Diels et Gilg were compared.   Method   Tubers from Huaiyu 1 (HY1) and Huaiyu 2 (HY2) were used for the transcriptome determination by PCR analysis.   Result  HY1 and HY2 had clean reads of 42 311 662 and 41 411 202, respectively, and no less than 95.75% of Q30 base. Their transcription factors basically belonged to the MYB-superfamily, bHLH, AP2/ERF, NAC, C2C2, WRKY, etc. The paired values of FPKM in HY1 and HY2 were between 0 and 2; the expression densities, between 0 and 0.7; the number of commonly expressed genes, 22 367; and, the number of uniquely expressed genes in HY1, 18 196, while 8 137 in HY2. The correlation between the expressions of the two had a coefficient of 0.913, and that between the samples was high. There were 12 199 differentially expressed genes between the two, with 3 551 upregulated and 8 648 downregulated in HY2 as compared to HY1. The GO enrichment analysis showed that the differential genes were mainly annotated into photosynthesis, light harvesting in photosystem I, photosynthesis, light harvesting, chlorophyll metabolic process, protein-chromophore linkage, generation of precursor metabolites and energy, chlorophyll biosynthetic process, response to oxidative stress, alpha-amino acid metabolic process, photosynthesis, plastoglobule, photosystem I, photosystem II, plastid nucleoid, photosystem, chlorophyll binding, monooxygenase activity, iron ion binding, heme binding, lyase activity, etc. Whereas, the KEGG enrichment analysis indicated the differential genes to be mainly annotated into photosynthesis-antenna proteins, ribosome, glyoxylate and dicarboxylate metabolism, phenylpropanoid biosynthesis, stilbenoid, diarylheptanoid and gingerol biosynthesis, flavonoid biosynthesis, photosynthesis, carbon fixation in photosynthetic organisms, glycine, serine and threonine metabolism, plant hormone signal transduction, glutathione metabolism, pyruvate metabolism, phenylalanine metabolism, circadian rhythm-plant, flavone and flavonol biosynthesis, cysteine and methionine metabolism, cyanoamino acid metabolism, carotenoid biosynthesis, alpha-linolenic acid metabolism, porphyrin and chlorophyll metabolism, and other metabolic pathways.   Conclusion  The differentially expressed genes related to flavonoids synthesis, such as stilbene synthase, leucoanthocyanidin dioxygenase, CHI protein, chalcone synthase 2, flavanone 3-hydroxylase and lleucoanthocyanidin reductase 1 and flavonoid 3’- hydroxylase gene were upregulated in HY2, while chalcone synthase, flavonol synthase and flavonoid 3’, 5’-methyltransferase downregulated. The variations apparently resulted in the differences shown on the total flavonoid content between the HY1 and HY2 tubers.
Inhibitory Effects of Ailanthus altissima Alkaloids on Pathogenic Fungi
LI Yongchun, ZHAO Meirong, ZHANG Zhi
, Available online  
Abstract:
  Objective  Possibility of applying Ailanthus altissima alkaloids for controlling 5 plant pathogenic fungi was studied.   Method  Crude ethanol extract of alkaloids from the bark of A. altissima was obtained. Extract was separated using the vacuum liquid chromatography. Antifungal activities of the crude alkaloids and the elution fractions on Fusarium oxysporum f. sp. cucumerinum, F. graminearum, F. oxysporum f. sp. benincasae, Botrytis paeoniae, and Gloeosporium sp. were determined according to the fungal mycelium growth rate.   Result  The crude ethanol-extracted alkaloids and canthin-6-one fraction exhibited inhibitory effects on the tested fungi, except B. paeoniae. The antimicrobial effects were particularly strong on Gloeosporium sp. and F. graminearum and especially toxic toward the latter with EC50 values of 0.31 mg·mL−1 by the crude alkaloids and 20.32μg·mL−1 by canthin-6-one.  Conclusion  The ethanol-extracted alkaloids from A. altissima barks could inhibit the growth and/or be lethal toward a variety of plant pathogenic fungi. Although the antifungal effect was not as potent as that of canthin-6-one, the crude extract per se appeared promising to be a new source for the development of a plant-based biocontrol agent.
Resistant identification and evaluation of 590 soybean varieties to anthracnose
SHI Niuniu, DU Yixin, HE Yanqin, RAUN Hongchun, TENTG Zhenyong, GAN Lin, LIAN Jinfan, YANG Zhonglu, CHEN Furu
, Available online  
Abstract:
  Objective   The objective of this study was to screen varieties resistant to soybean anthracnose through resistance identification, which will provide scientific basis for screening resistant varieties and disease resistance breeding.   Method   For this purpose, the resistance of 590 soybean varieties to anthracnose from 2011 to 2019 was determined by artificial inoculation.   Result   The results showed that the number of resistant varieties (R), medium resistant varieties (MR), medium susceptible varieties (MS), susceptible varieties (S) and high susceptible varieties (HS) was 70, 143, 219, 148 and 10, respectively, with ratio of 11.68%, 24.24%, 37.12%, 25.08% and 1.69%, respectively. Generally, the resistant ratio (RR) ranged from 13.64%~68.00% in different years. Among these, RR in 2013 was the highest, with value of 68.00%, followed by 52% in 2012, while RR in 2016 was the lowest, with value of 13.64%. Moreover, RR ranged from 5.89%–69.23% in spring soybean group of tropical and subtropical areas (S1), summer soybean group of tropical and subtropical areas (S2), fresh soybean spring sowing group (S3), fresh soybean summer sowing group (S4), spring soybean group of Yangtze river basin (S5), new soybean varieties group of fujian province (S6) and other new variety groups (S7). Across 7 groups, RR in S2 group appeared the highest, with 69.23%, followed by 57.69% in S4 group, whereas RR in S3 group demonstrated the lowest, with 5.89%.   Conclusion   The identification results showed that resistant and moderately resistant varieties accounted for 36.10% among 590 soybean varieties. There were no varieties with high resistant. And there was lack of varieties with good resistance to anthracnose in fresh soybean spring sowing group.
Research Progress on HD-ZIP Transcription Factors in Response to Pathogenic or Abiotic Stresses
Liu jiao, Shuai peng
, Available online  
Abstract:
Adverse external conditions commonly affect plant growth and development which directly or indirectly cause decline on crop yield and even death of the plants. The homeodomain-leucine zipper (HD-ZIP) transcription factors have been known to involve in stress responses of plants. Belonging to the homeobox (HB) protein family, the factors are unique to plants and tightly connected by the highly conserved HD and ZIP. The formation of protein dimers mediated by the LZ domain allows HD to bind to the target DNA and regulate the expression of the target gene. The HD-ZIP transcription factors not only play an important role in regulating plant growth and development but also in responding to external stresses. This article focuses on the published reports of recent studies concerning the roles of the 4 subfamilies Ⅰ-Ⅳ of the HD-ZIP transcription factors in response to pathogenic attacks and/or abiotic stresses such as drought, salt, extreme temperature, wounding, low R/FR light, and heavy metals. Through the internal molecular response mechanisms initiated by HD-ZIP, a plant could ward off the imposed adversities. With an in-depth understanding of the functions, means to improve the growth and stress resistance of plants could be realized.
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
, Available online  
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.