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Construction of Yeast Two-hybrid Bait Vector and the Screeningof Proteins Interacting with BsMYB62 in Begonia semperflorens
LIU Jing, WANG Yang, YAO Kexin, QU Ying, WANG Ruibo, LIU Shengnan, LI Yonghua, ZHANG Kaiming
[Abstract](0) [FullText HTML](1) [PDF 959KB](0)
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  Objective  Both low temperature and high light are important triggers in inducing anthocyanin biosynthesis in Begonia semperflorens. Selecting from the transcriptomes of B. semperflorens treated with low temperature and high light, we cloned the full length cDNA of an MYB transcription factor which was named as BsMYB62 (Gene ID: MT560845 ). We then analyzed the biological information, and selected the potential interaction proteins of BsMYB62. Our results are expected to elucidate function of BsMYB62.  Method  After designing the primers and cloning the full length cDNA, the gene and protein sequence of BsMYB62 were analysized using bioinformatics softwares. We constructed a pGBKT7-BsMYB62 bait vector, and tested the toxicity and auto-activation of the bait vector. Potential interaction proteins were obtained from the yeast-two hybrid library using the Mating method. Results are as follows.  Result  The full length cDNA of BsMYB62 is 801 bp. The pGBKT7-BsMYB62 is non-toxic to the host yeast, and may significantly inhibit the BsMYB62 protein auto-activating in SD/-Trp-Leu-His-Ade medium including 45mM of 3-AT. There are 9 kinds of potential interaction proteins, including XTH9, LHB1B2, EBS7, PSI-F, UBE2, FDH, RBCS1A, FP6 and BCA.  Conclusion  It is speculated that the BsMYB62 protein of B. semperflorens interacts with the above nine types of proteins in response to low temperature and high light stress.
Pathogen Identification and Biological Characteristics of Chimonanthus Salicifolius Leaf Spot
PENG Chengbin, WANG Zerong, RUAN Junfeng, WEI Rifeng, XUE Lan, CHEN Meixia, LIU Wei
[Abstract](1) [FullText HTML](0) [PDF 569KB](0)
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  Objective   Isolations, classifications, and biological characteristics studies of pathogens of Chimonanthus Salicifolius leaf diseases.   Method   The pathogen of leaf spot disease were isolated from sick trees leaves from Shouning county in Fujian Province with in vitro tissue culture method. The morphology and molecular identification was carried out. The biological characteristics and optimal growths conditions of mycelia including temperature, pH, light, carbon source, nitrogen source and lethal temperature were performed.   Result   Based on morphology features and rDNA ITS and TUB sequence analysis, the pathogenic fungus caused leaf spot was identified as identified as AscomycetesPleosporalesEpicoccumE. sorghinum. The biological characteristic resluts show that the pathogen can grow at 5~30℃; and the optimum temperature is 25℃; the optimal pH range of mycelium growth is pH 5~9; the strain can grow in the 11 kinds of carbon sources medium and the optimal carbon sources were sucrose; among 8 different nitrogen source medium, peptone was the optimal one; In addition, the growth of mycelia was not influenced by light and dark alternations; and the lethal temperature of mycelia was 52℃.   Conclusion   In the paper, the major pathogen causing leaf spot disease in C. Salicifolius from Shouning county in Fujian province was E. sorghinum, which exhibited high resistances, strong growth abilities and serious pathogenicity.
Analysis on Aroma Components of Grape Benni fuji by HS-SPME-GC-MS
LI Kai, SHANG Jiayin, SU Hong, TIAN Shufen, HUANG Jianquan, ZHANG Na, WANG Dan, WANG Chaoxia
[Abstract](0) [FullText HTML](0) [PDF 682KB](0)
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  Objective   The aroma components of grape Benni fuji grown in Tianjin were detected to study the active aromatic components and aroma characteristics of the grape.   Method   Headspace solid phase microextraction (HS-SPME) and gas chromatography-mass spectrometry (GC-MS) were used to extract and detect the aroma of grape Benni fuji. Internal standard - standard curve method was used for quantitative analysis. The aroma components and aroma characteristics of grape were analyzed by combining the aroma activity value and aroma profile.   Result   Quantitative results showed that a total of 39 aroma components (total concentration 9737.05 μg·L−1) were identified and quantified, esters were the dominant component, followed by alcohols. Ethyl acetate, 1-hexanol, trans-2-hexene-1-ol, ethyl butyrate and trans-2-hexene-1-al were the five dominant aroma components, whose total concentration (9461.79 μg·L−1) accounts for 97.17% of the total concentration detected. Ethyl acetate concentration (9036.53 μg·L−1) was the most outstanding, accounting for 98.53% and 92.81% of the ester concentration and the total concentration respectively. Odor activity value results showed that aroma of grape Benni fuji was mainly contributed by 12 kinds of active aroma components, including ethyl butyrate, ethyl-2-methylbutyrate, ethyl hexanoate and ethyl isobutyrate. The visualized aroma profile showed that aroma of grape Benni fuji was composed of flowers, fruits, plants, fats, chemicals and spices flavor, and dominated by fruits flavor.   Conclusion   The aroma components of grape Benni fuji grown in Tianjin was mainly composed of esters, alcohols and aldehydes, and esters were the dominant components, contributing the outstanding fruits flavor.
Identification of oil palm polyamine synthesis gene and expression analysis under cold and dehydration stress
JIN Longfei, YIN Xinxing, ZHANG Anni, CAO Hongxing
[Abstract](0) [FullText HTML](0) [PDF 1224KB](0)
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  Objective   To provide useful information and resource for oil palm resistance varieties breeding, we investigate the roles of polyamines synthase on oil palm growth, development and abiotic stress response.   Method   15 polyamines synthase genes were identified from oil palm genome. Bioinformatics and expression pattern were analyzed.   Result   The amino acid number was between 318 to 720; the molecular weight was between36.00 and 46.43 kDa, the isoelectric point was between 4.78 and 6.31, the instability index was between 29.26 and 48.73, the aliphatic index was between 76.46 and 94.41, the gravy was between-0.315 and 0.082, the exon numerous was between 1 to 11. Polyamine synthesis genes contain specific conserved protein motifs and conserved domain. Phylogenetic analysis indicated that polyamine synthesis genes can be divided into three families; and EgADCs were closely related to rice, EgACLs and EgSPDSs were closely related to data palm, EgSPMSs were closely related to maize. A large number of phytohormone responses, stress responses, light responses cis-acting elements were identified on promoters. Gene expression analysis showed that polyamine synthase genes were expressed in different tissues of oil palm. Polyamine synthase genes were induced by cold and dehydration stress.   Conclusion   EgSPDS1, EgSPDS2, EgSPMS1, EgADC1, EgADC2, EgACL5-2, EgACL5-3, EgSAMDC3-1, EgSAMDC4-1, EgSAMDC4-2 and EgACL5-1 were induced by cold stress; EgSPDS1, EgSPDS2, EgADC1, EgADC2, EgSAMDC4-1 EgSAMDC4-2 and EgACL5-1 were induced by dehydration stress.
Physiological and biochemicalmechanism of inhibition effect of low temperature on flower bud differentiation of Narcissus tazettavar. chinensis
LI Lin, LI Tingting, YANG Kun, ZHANG Zhizhong, WU Jinghua
[Abstract](0) [FullText HTML](0) [PDF 830KB](0)
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  Objective  In this paper, we study the physiological mechanisms of the low temperature suppression on flower bud differentiation of Narcissus tazetta var. chinensis, which provides theoretical support for the flowering regulation of Chinese narcissus.  Method  The 3-year-oldChinese narcissus bulb were treated with 15℃ low temperature and room temperature. The protective enzyme activity, soluble protein and carbohydrate content in scales were measured at different stage.  Results  In the control group with the room temperature treatment, the content of both superoxide anion (O2) and hydrogen peroxide (H2O2) first increases, then decreases, and finally increases. The activity of Superoxide dismutase (SOD) remains at a high level, while the peroxidase (POD) activity is at a lower level. In addition, the activity of catalase (CAT) increases significantly at a late stage. Also, the content of sucrose and starch decreases in the floret primordium, whereas the content of α-amylase and soluble sugar stays in a high level at the same time. With the low temperature treatment, the content of both O2 and H2O2 is higher than the one in the control group. The change in the SOD activity is similar to that for the control group. Furthermore, the extent of POD and CAT activity in stamen primordium is significantly higher in the lower temperature treatment. The content of soluble sugar, sucrose, starch, as well as the extent of α-amylase activity in the low temperature treatment are relatively lower at each sample point, compared with the control group.  Conclusion  The contents of soluble sugar, sucrose and starch played an important role in the process of flower bud differentiation. The accumulation of carbohydrates was greatly affected by low temperature treatment at 15°C, which further affected flower bud differentiation. This effect was most intense during the floret primordium stage.
Response of rhizosphere soil bacterial community maize on acid soil to straw incorporation and soil conditioner
ZHANG Hui, LIN Chenqiang, CHEN Longjun, CHEN Jichen
[Abstract](0) [FullText HTML](0) [PDF 995KB](0)
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  Objective   The technology of high-throughput sequencing on Illumina Miseq platform was adopted to the changes of bacterial community composition and diversity of maize rhizosphere soil bacterial community treated by no chemical fertilizer (CK), single chemical fertilizer (T1), straw mulching with chemical fertilizer (T2), and soil conditioner with chemical fertilizer (T3).   Methods   Using the Illumina Miseq sequencing platform sequencing of four groups of treatment of soil samples, based on the analysis of the second generation of high throughput technology no chemical fertilizer (CK), single chemical fertilizers (T1), chemical fertilizer with straw returned (T2), chemical fertilizers with soil conditioner (T3) of maize rhizosphere bacterial 16S rRNA genes in V3, V4 area of diversity index, richness and community composition and structure.   Results   The results showed that the dominant bacteria (relative abundance > 10%) were Proteobacteria, Chloroflexi and Actinobacteria. There were altogether 4, 183 dominant genera (relative abundance > 1%). The richness index of T1 bacteria (ACE index and Chao1 index) was lower than that of CK, which decreased by 17.65% and 17.88% respectively. The bacteria richness index (ACE index and Chao1 index) in T3 treatment were higher than those in CK treatment, with an increase of 14.52% and 14% respectively. The stratified cluster diagram showed that, at the level of bacteria genera, the CK, T1 and T2 treatment soil samples had higher similarity in bacterial community structure, while T3 and the other 3 treatments had greater difference in bacterial community structure.   Conclusion   Fertilizer with soil conditioner influence to soil bacteria community structure is greater than the single fertilizer and chemical fertilizer with straw counters-field processing, single fertilizer reduces the richness of rhizosphere bacteria, chemical fertilizer with soil conditioner can significantly improve the rhizosphere bacteria community richness, applying soil conditioner to improve acidic soil pH had the greatest influence on maize rhizosphere bacteria community.
Relationship between soil properties and quality of Duobei tea in Puding
GUO Jianjun, WANG Jinjin, MA Qiang, ZHOU Yi, LI Huihui, WU Guohua
[Abstract](0) [PDF 0KB](0)
Abstract:
  Objective  The objective of this research was to study the relationship between soil properties and tea quality of Duobei tea, so as to provide theoretical basis for improving nutrient management in tea plantation and subsequent tea quality.  Method  Soil samples were collected from nine different planting areas of Duobei tea. The metal content in the soil and its pH value, organic matter, available phosphorus, alkali hydrolysis nitrogen, available potassium and the activities of sucrase, urease, and catalase was determined. Correlation analysis with the nutritional composition and sensory evaluation and antioxidant activity of Duobei tea.  Result  The results showed that the content of organic matter and available phosphorus in the soil of 9 different planting areas all reached the first-grade fertility standard, and the available nitrogen and available potassium were generally low. Especially the 7# block field, include with 45.57 g·kg−1 organic matter, 46.90 mg·kg−1 alkali hydrolyzed nitrogen, 14.81 mg·kg−1 available phosphorus, 157.00 mg·kg−1 available potassium, and the pH was 4.68. Combined the sensory evaluation and nutrient composition analysis of tea, the quality of Duobei tea showed the characteristics of "tender, fresh, thick and mellow". Among them, the quality of 1#, 2# and 7# tea was better, and the 7# tea was the strongest antioxidant capacity in vitro. Correlation analysis showed: the higher soil organic matter, alkali hydrolysis nitrogen content, urease and invertase activities had a positive effect on the quality of tea nutrients and various sensory evaluation factors; The increased in soil pH, higher available phosphorus content and higher catalase activity had adverse effects on the quality of tea nutrients and various sensory evaluation factors.  Conclusion  In general, to improve the quality of Dobe tea, the management measures should be "addition of nitrogen and potassium to control phosphorus, and increase of organic matter".
Drought-tolerant screening during seed germination and genetic diversity analysis of mungbean germplasms
HUANG Nianying, ZHU Zhenzhen, LIU Changyan, LIAO Fangli, LI Li, CHEN Hongwei, LIU Liangjun, HAN Xuesong, WAN Zhenghuang, SHA Aihua
[Abstract](0) [PDF 0KB](0)
Abstract:
  Objective  Evaluating the drought tolerance of 56 mungbean accessions, and understanding their genetic bases.  Method  10% concentration of mannitol solution was used to simulate drought stress on the 56 germplasm resources, and the relative germination rate, relative germination potential and relative root length etc. were used as drought tolerance evaluation indicators. The drought tolerance of mung beans during germination were evaluated by conbing of the principal component analysis, membership function analysis and multivariate analysis methods. The genetic base of 56 germplasms was investigated by cluster analysis based on SSR.  Result  Drought stress had different effects on 10 drought tolerance evaluation indexes in germination stage. Principal component analysis showed that the contribution rates of the first three comprehensive indicators were 57.87%, 16.42%, and 11.42% with the cumulative contribution rate of 85.71%, which could represent the variation information of 10 evaluation indicators. According to the comprehensive evaluation value D calculated by principal component analysis, the drought tolerance of 56 germplasms was evaluated. 2 high tolerance, 10 drought tolerance, 30 medium tolerance, 13 sensitive and 1 high sensitive germplasm were identified. the 56 accessions were divided into three main groups based on SSR cluster analysis. The first group contained 25 germplasm, which showed relatively drought tolerance. The second group contains 26 germplasm resources showing different degrees of drought tolerance due to complex genetic basis. The third group contains 5 germplasms, which are prone to sensitivity.  Conclusion  The drought tolerance and genetic characteristics of 56 mung bean germplasms at germination stage were determined. These results laid a foundation for drought tolerance breeding of mung bean by utilization of these germplasms.
The Expression Analysis of Panicle Development And Cell Cycle Related Genes in Clustered Spikelets Rice
CHEN Liping, JIANG Jiahuan, ZHENG Yanmei, ZHU Yongsheng, WANG Yingheng, LIN Qiang, XIE Hongguang, LUO Xi, CAI Qiuhua, XIE Huaan, ZHANG Jianfu
[Abstract](0) [PDF 0KB](0)
Abstract:
  Objectives  The increase of grain number of per panicle directly improves the yield of rice and the clustered spikelets increase the grain number of per panicle and the density of panicle. The objectives of this paper are to investigate the expression differences of OsCl-6 and other panicle development related genes and cell cycle related genes between the clustered spikelets variety Neijiang P164 and single spikelet rice variety 9311, to explore the guidance for rice production by regulating the expression of specific genes in rice varieties, and to provide a theoretical basis for the breeding of high-yield rice varieties.  Methods  The young panicles of the clustered spikelets variety Neijiang P164 and single rice variety 9311 planted in the field of net room were taken for RNA extraction, qRT-PCR analysis and related data analysis, respectively.  Results  The results of qRT-PCR showed that the expression level of OsCl-6 in young panicles at different stages of development in Neijiang P164 was higher than in 9311.. The expression levels of cell cycle dependent kinase genes in 9311 was higher than in Neijiang P164, while the expression levels of class B cyclins in Neijiang P164 was higher than in 9311. The expression levels of panicle development related genes such as LAX1, WOX3 and RFL in Neijiang P164 was lower than in 9311, but MFS1 and CL-4 in Neijiang P164 were higher in 9311.  Conclusions  There were significant differences in the expression of panicle development related genes, including OsCl-6 and B-type cyclins in the Naijiang P164 and 9311, OsCl-6 may be up-regulated to inhibit the development of secondary branches in rice, which will provide a reference for the further study of the relationship between panicle development related genes and the molecular mechanism on how OsCl-6 regulate the formation of the clustered spikelets.
Heterologous expression and possible temperature regulation mechanism of the synthesis gene clusters of prodigiosin from Serratia marcecens
LIU Fangchen, JIA Xianbo, WU Liangquan, FANG Yu, ZHAO Ke, LIN Junjie, CHEN Longjun, ZHANG Hui, LIN Chenqiang, CHEN Jichen
[Abstract](0) [PDF 0KB](0)
Abstract:
  Objective  To construct a heterologous expression strain with the prodigiosin synthesis gene cluster from Serratia marcecens FZSF02, and understand the potential temperature regulation mechanism of the synthesis of prodigiosin.  Methods  Three heterologous expression Escherichia coli strains carrying different promoters were constructed, and real-time fluorescent quantitative PCR (RT-qPCR) was used to determine the transcriptional differences of pigA, pigF and pigN in pig gene cluster at 37 ℃ and 28 ℃.  Results  At a low temperature of 28 ℃, all three different promoters can initiate the expression of the pig gene cluster in Escherichia coli, prodigiosin was also produced in these three recombinant Escherichia coli strains at 27 ℃. Among them, the recombinant strain carrying T7 strong promoter could product a small amount of pigment at 37 ℃ after IPTG induction, while the other strains could not product pigment at 37 ℃. RT-qPCR results showed that transcription levels of the above genes were down-regulated at 37 ℃ compared with 28 ℃ in the S.marcecens FZSF02.  Conclusion  The reason why the stain S. marcecens FZSF02 can product prodigiosin at 28 ℃ but can't product that at 37 ℃ might be that, on the one hand, the transcriptional levels of the genes of the pig gene cluster were down-regulated at higher temperatures; on the other hand, the activities of one or several key enzymes encoded by the pig clusterfor the biosynthesis of the precursor MBC and MAP were inhibited at a relatively higher temperature (such as 37 ℃).
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
[Abstract](17) [FullText HTML](14) [PDF 608KB](2)
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
[Abstract](45) [FullText HTML](18) [PDF 1252KB](6)
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
[Abstract](27) [FullText HTML](10) [PDF 575KB](1)
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  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.
The research for genomic DNA extraction methods and authenticity identification in Clematis florida Thunb. var. plena D.Don and its confused
HE Shulan, Li Yongning, ZHU Furong
[Abstract](0) [PDF 0KB](0)
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  Objective  To establish a rapid and economical method for extracting genomic DNA from Clematis florida Thunb.var.plena D.Don, and to identify the authenticity of this Chinese herbal medicine accurately by molecular pharmacognosy, so as to lay a foundation for further development and application of this characteristic Chinese herbal medicine in Fujian Province.  Method  Using CTAB method, modified CTAB method, Kit method, SDS method, high salt and low pH method to extract genomic DNA of the primitive plant leaves of Clematis florida Thunb.var.plena D.Don. The quality and integrity of DNA were detected by agarose gel electrophoresis. The genomic DNA was extracted by the optimized method. Its DNA and its confused DNA were sequenced bidirectionally to evaluate the identification effect of this molecular pharmacognosy method.  Result  The purity of DNA extracted by modified CTAB method was high, and the bands obtained by SSR primer amplification and ITS2 sequence amplification were clear and bright, which was the best method for extracting genomic DNA of Clematis florida Thunb.var.plena D.Don. The PCR amplification products of genomic DNA extracted by this method can be used for bidirectional sequencing, and then identify its source, so as to accurately distinguish it from common confused products.  Conclusion  The improved CTAB method established in this study can effectively extract the genomic DNA of Clematis florida Thunb.var.plena D.Don. The method is simple and has good DNA quality, which can be used for molecular pharmacognosy identification of Clematis florida Thunb.var.plena D.Don and confusion.
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
[Abstract](28) [FullText HTML](11) [PDF 1023KB](4)
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
[Abstract](56) [FullText HTML](27) [PDF 799KB](5)
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.
Identification of the pathogen of Bacterial Soft Rot of Paris polyphylla Smith var. chinensis (Franch.) Hara in Fujian Province
ZHENG Meixia, SU Hailan, XIAO Rongfeng, HE Guiqiang, ZHU Yujing
[Abstract](0) [PDF 0KB](0)
Abstract:
  Objective   The purpose of this study is to investigate the characteristics and harm of Paris polyphylla Smith var. chinensis (Franch.) Hara soft rot, isolate and identify the pathogen of soft rot, and provide evidence for the field control of the disease.   Method   In this study, the investigation of field disease and the collection of Paris polyphylla Smith var. chinensis (Franch.) Hara sample were carried out, and the samples were subjected to pathogen isolation and purification, morphological characteristics identification, 16S rDNA gene sequence characteristics and pathogenicity determination, respectively.   Result   The results showed that Paris polyphylla Smith var. chinensis (Franch.) Hara leaves, stems and tubers all have soft rot. Soft rot usually occured in the field which had poor ventilation and water accumulation. In the early stage of the disease, water-soaked necrosis and pungent odor appeared in the affected part, and then the internal tissue of the affected part was broken down into liquid. Morphological, molecular biological characteristics and pathogenicity determination of the pathogen showed that the pathogen was P. carowvorum.   Conclusion   P. carowvorum is the pathogen of Paris polyphylla Smith var. chinensis (Franch.) Hara soft rot. Paris polyphylla Smith var. chinensis (Franch.) Hara is easily infected by P. carowvorum in poorly ventilated and poorly drained environments.
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
[Abstract](23) [FullText HTML](6) [PDF 599KB](2)
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.
The study on tea plant cultivars resistance to green leafhopper based on PPIN
WANG Fen, PEI Huimin, WEN Di, CHEN Zhi, LI Jing
[Abstract](35) [FullText HTML](7) [PDF 760KB](6)
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.
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
[Abstract](24) [FullText HTML](9) [PDF 747KB](1)
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 then the SU gene was 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.
Cloning and Sequencing Sucrose Synthase Gene of Alpine Potato
HONG Senrong, DENG Yuqing, WU Hongting, CHEN Ting, GUO Shuzhen, YANG Yuxuan, CAI Hong, CHEN Ronghua
[Abstract](16) [FullText HTML](13) [PDF 1103KB](6)
Abstract:
  Objective   Sequence of the gene associated with the key enzyme in sucrose metabolism pathway of potatoes, sucrose synthase (SuSy), was determined for studying the biological function of the enzyme.   Method  The core fragment (PGSC0003DMG400002895, SuSy 4) of SuSy was screened from the transcriptome database on the plantlets of Alpine potatoes (Solanum tuberosum L. cv. Huaiyushan). The gene was cloned by RT-PCR, and its sequence analyzed using the bioinformatics method.   Result   The total length of cDNA of SuSy gene was 2 418 bp containing 45.08% of G+C. The hydrophilic protein had 805 amino acids with a molecular weight of 92 471.33 Da and an isoelectric point of 5.87. Its secondary structure was 45.84% of alpha helix, 15.16% of β -extended strands, and 39.01% of random coils. The β-lamellae and α-helix were found throughout the entire protein but appeared as irregular, curl, extension chains at the C- and N-terminals. The tetrameric gene mainly existed in the cytoplasm, mitochondria, and chloroplast. It closely related to those of S. lycopersicum, S. pennellii, S. chilense, S. tuberosum, Capsicum annuum, and C. baccatum in evolution, especially, S. tuberosum.   Conclusion  The SuSy gene in the Alpine potato of Huaiyushan had the characteristic structure of a typical sucrose synthase. The amino acid and nucleic acid sequences were highly conserved and homologous to the similar species. The findings would be of great significance for further understanding on the biological function of the enzyme.
Genetic Diversity on Phenotypes of Blumea balsamifera Germplasms
XIAO Yongfeng, HUANG Mei, YU Fulai, CHEN Zhenxia, LIAO Li, PANG Yuxin
[Abstract](15) [FullText HTML](1) [PDF 657KB](0)
Abstract:
  Objective  The diversified phenotypic characteristics of Blumea balsamifera (L.) DC. germplasms were classified to facilitate the breeding program.   Method  Nine quantifiable and 13 quality phenotypic traits of the plant were used to describe the 159 germplasm samples of B. balsamifera. Genetic diversity, correlation, principal component, and cluster analyses were applied for the study.   Result   The genetic diversity on phenotypic characteristics of the collected germplasms was rich. Among the quantifiable traits, plant height had the highest genetic diversity index of 2.072; and, number of flowering branches topped the coefficient of variation at 32.76%. The crown breadth of the plants was found significantly correlated with the height, leaf width, petiole length, length of flower branch, and opening angle of flower branch (P<0.01). On the quality traits, the highest genetic diversity index of 1.201 was the leaf shape. And, the color intensity of anthocyanin in leaf vein and that in main stem, as well as, those in leaf edge and petiole were significantly correlated. The cumulative contribution rate of the first eight principal components reached 64.32%. They included the factors associated with yield, color-intensity, leaf smoothness, leaf edge, leaf shape, leaf greenness, and flower-branch angle. Based on the phenotypic traits, the sum of squares deviations method using the genetic distance of 10 on the germplasms divided the 159 varieties into Group Ⅰ of 39 that accounted for 24.53% of the total, Group Ⅱ of 38 that accounted for 23.90% of the total, and Group Ⅲ of 82 that accounted for 51.57% of the total.  Conclusion   Genetically, the phenotypic characteristics of the 159 B. balsamifera germplasms appeared richly diversified. The leaf width of the plants could be the most outstanding trait for breeding selection of high-yield B. balsamifera varieties.
Effects of Rice Straw Biochar on Water-soluble Cd in Three Flooded Different Types of Soil
ZHANG Jiakang, ZHUAN Yalin, ZHANG Liwen, LIN Guiquan, LIN Yongchong, LI Honghong
[Abstract](33) [FullText HTML](13) [PDF 1153KB](7)
Abstract:
  Objective   To investigate the effect of biochar made from rice straws on cadmium (Cd) immobilization in different types of soil under flooding.   Method   An in-lab experiment on 3 Cd-added different types of soil with or without a 5% addition of biochar made from rice straws was conducted under varied durations of flooding. The types of soils used were yellow soil, paddy soil derived from quaternary red clay, and brown soil.   Result   The biochar reduced pH and redox potential (Eh) but increased conductivity of the soils on the first day after flooding. As the flooding persisted, Eh in the soils reduced continuously, but the rate declined in the presence of the biochar. During the initial stage of flooding, the Cd contents in the soils were highest in the yellow soil at 272.5 μg·L−1 followed by the paddy soil at 23.48 μg·L−1, while the brown soil at 1.44 μg·L−1 being the lowest. The Cd reductions by 31.66% in the yellow soil, 75.04% in the paddy soil, and 66.67% in the brown soil were attributed to the added biochar. Under prolonged flooding, the Cd in the yellow and paddy soils gradually decreased even without the biochar addition. In 30d, the reductions were 89.34% on the yellow soil and 76.53% on the paddy soil. In comparison, the addition of the biochar brought about 85.41% Cd reduction on the yellow soil and 37.03% on the paddy soil. After 30d of flooding, the biochar out-performed control with the CaCl2-Cd contents in the yellow, paddy, and brown soils lowered by 17.3%, 56.3%, and 12.4%, respectively.   Conclusion  By adding 5% of the rice straw biochar, the water-soluble Cd in the 3 different types of soil could be significantly reduced. Prolonged flooding made the effect less pronounced, and the biochar immobilization of Cd appeared more effective in the paddy soil than the other two soil types.
Correlation between SlSIP1L12 Expression and Seed Germination of Tomato Plants
CUI Baolu, CHEN Guoping
[Abstract](14) [FullText HTML](9) [PDF 1070KB](1)
Abstract:
  Objective  Role of SlSIP1L12, an SIP1 subfamily gene of trihelix transcription factors, played in tomato seed germination was investigated.   Method  Expressions under and responses to phytohormone and abiotic stress of SlSIP1L12 were tested by RT-PCR. The RNAi technology was used to suppress the SlSIP1L12 expression to reveal its function on the seed germination. ELISA was applied to detect the endogenous ABA contents in the seeds.   Result   (1) The length of SlSIP1L12 was 1125bp in AC++ that encoded 374 amino acids. The phylogenetic analysis confirmed that SIP1 genes had indeed evolved in AC++. (2) SlSIP1L12 was primarily expressed in the stems and mature leaves, and secondly, in the flowers and fruits. (3) The suppressed expression of SlSIP1L12 could be artificially induced by ABA or dehydration, which implied a close relationship between the gene and ABA. (4) The germination of the SlSIP1L12 transgenic seeds was faster and grew to longer radicles than control. (5) ABA contents were significantly reduced in the transgenic seeds 7d after germination.   Conclusion  The downregulated SlSIP1L12 that stimulated germination of tomato seeds was postulated to be closely associated with the ABA response of the plant.
Physiological and Biochemical Properties of Agaricus bisporus in Shaking Flask Culture
DAI Jianqing
[Abstract](19) [FullText HTML](9) [PDF 1161KB](0)
Abstract:
  Objective   Mycelial growth of Agaricus bisporus W192 in a shaking flask was monitored to determine viability of the culture method for scale-up application.   Method   Physiological and biochemical properties of A. bisporus mycelia biomass in the liquid culture including the number and diameter of mycelium pellets, pH of fermentation broth, content of reducing sugar and amino nitrogen as well as the activity of extracellular carboxymethyl cellulase, amylase, acid protease, and laccase were monitored for the analysis.   Result   On the 8th day of culture, the fungal biomass reached a maximum at 9.7 mg·ml−1, the greatest number of mycelium pellets of 880 CFU·mL−1, the largest average diameter of mycelium at 0.809 mm, the peak reducing sugar and amino nitrogen contents at 5.228 and 0.079 mg·ml−1, respectively, and the highest activities of carboxymethyl cellulase, amylase, and laccase of 0.69 , 2.11 , and 15.02 U, respectively. The acid protease activity of 2.93U maxed on the 10th day. By transferring the mycelia in a simulated liquid fermentation tank after 8d of culture, 115 CFU·mL−1 of mycelium pellets and 7.56mg·mL−1 in biomass were obtained.   Conclusion   The mycelial growth of A. bisporus in the liquid shaking flask culture correlated with some of selected physiological and biochemical indices. The established method was deemed applicable for tank fermentation in propagating A. bisporus for large scale production of the mushroom.
Isolation of Novel Goose Astrovirus, FJ-NP
LIN Yusheng, JIANG Jinxiu, ZHANG Jingpeng, YOU Wei, HU Qilin
[Abstract](16) [FullText HTML](2) [PDF 837KB](0)
Abstract:
  Objective  To isolate a previously unknown pathogen that caused an outbreak of highly lethal disease in a geese farm at Nanping, Fujian in August 2019.   Method  Three clinical samples from the diseased geese, which showed symptoms of the infectious disease characterized by gout with an incidence rate as high as 40% that resulted in a mortality rate of 80% on 5- to 20-d-old geese, were collected and tested in laboratory for pathogen identification.   Result  The PCR results confirmed the existence of novel goose astrovirus in all three samples, but no bacteria were isolated from medium culture. Inoculating the virus into the 11-d-old goose embryos did not induce death but showed dotted hemorrhage on the body skin and hemorrhage on the kidneys and lungs. The allantoic fluid and tissue specimens collected for PCR were tested positive as novel goose astrovirus, not other viruses commonly known to infect geese. The isolated virus was named FJ-NP, and its partial ORF2 genes showed a close relationship with the strains found in Anhui (i.e., GD AHAU2, AHAU3, and AHAU5), in Heilongjiang (i.e., AstV-Goose-2018-HLJ01) and in Henan (i.e., AstV-HN02-Goose-1119-18, AstV-AH02-Goose-0715-18, AstV-HB02-Goose-0310-19, and AstV-HN03-Goose-0402-19) but far from the Hubei, Beijing or Fujian strains.  Conclusion  This study successfully isolated a strain of novel goose astrovirus, and performed a genetic evolution analysis on its partial ORF2 genes.
Response Surface Optimization on Subcritical Water Extraction of Polysaccharides from Pitaya Stems
LI Guosheng, HUANG Xiuyin, BAI Xinpeng
[Abstract](0) [FullText HTML](1) [PDF 967KB](0)
Abstract:
  Objective   A subcritical water extraction method was optimized by response surface experiments to extract polysaccharides from pitaya stems.   Method  On the basis of a single factor test, pitaya stems were extracted using phenol-sulfuric acid to determine the polysaccharide content. To optimize the subcritical water extraction process assisted by ultrasonic pretreatment, water temperature, extraction time, liquid-to-material ratio, and pH were used against polysaccharide extraction rate in a response surface experiment.  Result  The optimized processing conditions included the applications of water at pH 5.9 and 144 ℃ with a liquid-to-material ratio of 31:1 (mL:g) to extract for 19 m. A polysaccharide extraction rate of 26.47% was achieved.   Conclusion  The established method appeared adequate for polysaccharide extraction from pitaya stems, and the mathematical model obtained be used to analyze and predict the process parameters.
Visual LAMP for Salmonella Detection on Soil for Vegetable Farming
LYU Xin, LIU Lanying, CHEN Lihua, LI Yueren
[Abstract](7) [FullText HTML](5) [PDF 726KB](3)
Abstract:
  Objective  A visual loop-mediated isothermal amplification (LAMP) technique for rapid detection of Salmonella in soil was developed to facilitate the prevention and control of the pathogens in vegetable fields.  Method  Based on the sequence of the invasion protein A (invA) of Salmonella, LAMP primers were designed and reaction system optimized. Specificity and sensitivity of the methodology were challenged by artificial inoculation and verified by the National Standard Detection Method for Salmonella Detection in Vegetable Cultivation Soil.   Result  The LAMP reacted positively on Salmonella but negatively on 6 non-Salmonella strains. It showed a minimum detection limit of 7 CFU·25 μL−1 and a sensitivity of 4×102CFU·g−1 on the artificially inoculated Salmonella. Compared with the standard method, the LAMP exhibited same testing accuracy.   Conclusion  The newly developed visual LAMP assay was rapid, accurate and sensitive for the detection of Salmonella in soil for vegetable farming.
Prokaryotic Expression and Purification of Tobacco Mosaic Virus Specific P54 Protein
LI Cong, TIAN Peijie, ZHANG Yu, ZHANG Deyong, LIU Yong, ZHANG Songbai
[Abstract](24) [FullText HTML](13) [PDF 1579KB](0)
Abstract:
  Objective   Pathogenicity and molecular mechanisms of tobacco mosaic virus (TMV), a typical member belonging to Tobamovirus of Vigaviridae that infect more than 400 species in 36 families of plants causing 20-30% reduction or complete loss on crop yield, were studied.   Methods   Sequence of the 1 425 bp TMV specific P54 protein was amplified by RT-PCR from the cDNA of Nicotiana tabacum var. Samsun NN infected with TMV and cloned into prokaryotic expressing plasmid, pEASY®-Blunt E1, followed by expressing in E. coli BL21 (DE3) by IPTG induction. Subsequently, the expression products were retrieved and purified by Ni-NTA chromatography and confirmed by western-blotting identification.   Results  The genome of TMV was a positive-sense single-stranded RNA of 6 400 bp that encoded one structural protein and two nonstructural proteins. The amplified 1 425 bp P54 was inserted in the virus replication associated P183 gene. The prokaryotic expressed recombinant P54 was insoluble. It was retrieved and purified to show a molecular weight of approximately 60 kDa and verified by means of western blot.   Conclusions   TMV P54 protein was successfully expressed and purified for further study on the devastating diseases on plants caused by TMV.
Rapid Species-specific PCR Identification of Bactrocera tau (Diptera: Tephritidae)
HUANG Zhen, HOU Youming, GUO Qiongxia, CHEN Shaoping
[Abstract](13) [FullText HTML](3) [PDF 786KB](0)
Abstract:
  Objective   A rapid method to detect Bactrocera tau (Walker) was established for inspection on produce to prevent potential introduction and spread of the invasive pest.   Method   Based on the species-specific PCR (SS-PCR) technology, B. tau as control and 20 common fruit flies of similar morphology were used to extract the genomic DNA templates, and the homology of the specimens checked with the COI mt DNA sequence. A rapid and accurate identification method applying the B. tau-specific primers NF404 and NR610 was designed and verified by PCR amplification and electrophoresis analysis.   Result  The newly developed SS-PCR detection method clearly amplified a single 207 bp band only on the target B. tau, not on any other fruit flies. Comparisons on the test results on the insects or body parts of all fruit flies with homogenous band sequence to B. tau validated the new methodology.   Conclusion  The newly established SS-PCR method exhibited high repeatability, specificity, and accuracy in detecting B. tau. It was considered applicable for the entry/exit and quarantine inspection on the invasive insect, larvae or dead body parts found on vegetables and fruits.
Microalgae for Effective Nitrogen and Phosphorus Removal from Aquaculture Effluence
JIA Wei, NIE Yilei, CHEN Hong, LUO Lijin, LE Zhanxian, ZHUANG Hong, ZHENG Junrong
[Abstract](112) [FullText HTML](73) [PDF 700KB](1)
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.
Preparation and Regeneration of Didymella bryoniae Protoplasts
MAO Jiancai, WANG Haojie, LI Junhua, ZHAI Wenqiang
[Abstract](15) [FullText HTML](11) [PDF 965KB](0)
Abstract:
  Objective  To better understand the pathogenicity of Didymella bryoniae on melons, this study aimed to establish an efficient means of preparing and regenerating the fungal protoplasts.   Method   Driselase at 20 g·L−1 and a lysing enzyme at 8 g·L−1 were combined and used for the protoplast preparations. A single-factor experiment was conducted to analyze the effects of age of mycelia, time and temperature of enzymatic digestion, rotational speed of culture vessel, type and concentration of osmotic stabilizer, and pH of medium on releasing of the protoplasts. Conditions for the subsequent protoplast regeneration in medium were also optimized.   Result  The highest yield of protoplasts of 9.65×107 cells·mL−1 was achieved using the mycelia cultured for 36 h to be enzymatically digested in a solution containing NaCl 0.7 mol ·L−1 as the osmotic stabilizer at 30 ℃ and pH 7.0 with 140 r·min−1 constant rotation for 4 h. The protoplast regeneration rate could reach up to 22.53% in a 0.5% agar SR culture medium but declined rapidly in 10 h and then leveled off in storage.   Conclusion  The protoplasmic preparation and regeneration methods were efficient and could materially aid the establishment and research on the genetic transformation of D. bryoniae.
Growth and Nutrient Content of Dendrobium officinale as Affected by Epulorhiza Symbiosis
WANG Weiying, ZOU Hui, DAI Yimin, LIN Jiangbo
[Abstract](14) [FullText HTML](9) [PDF 641KB](1)
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.
Cloning and Bioinformatics Analysis of Protein Phosphatase Genes MiSTPP1 and MiSTPP4 from Macadamia integrifolia
LI Jidong, YANG Xiangyan, CAI Yuanbao, LI Mu, ZENG Liming, HUANG Sijie, WU Fumin, LIN Yuhong, ZHENG Wenwu, HUANG Jinyuan
[Abstract](157) [FullText HTML](84) [PDF 967KB](3)
Abstract:
  Objective  Two new protein phosphatase genes, MiSTPP1 and MiSTPP4, from macadamia (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 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.
Effect of endogenous hormone content and gene expression related to hormone synthesis on tuber expansion of Bikeqi yam
AO Lan-ji-ya, JI Xiang, SHAO Ying, ZHAO Lingmin, ZHANG Yan-fang, HUO Xiu-wen
[Abstract](582) [FullText HTML](456) [PDF 693KB](14)
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.
2021, (1): 1-2.  
[Abstract](1) [FullText HTML](0) [PDF 369KB](0)
Abstract:
Animal Science
Codon Optimization and Expression of Bovine Pregnancy-related Glycoprotein-16 Gene
LIU Changbin, LU Chunxia, LU Shouliang, NI Jianhong
2021, 36(1): 1-8.   doi: 10.19303/j.issn.1008-0384.2021.01.001
[Abstract](7) [FullText HTML](5) [PDF 891KB](1)
Abstract:
  Objective   The bPAG16 gene related to bovine pregnancy was codon-optimized and synthesized to construct recombinant vector expressed in HEK293 cells for the development of a diagnostic tool on pregnancy of dairy cows.   Methods  Bioinformatics were applied to codon-optimize and synthesize the bPAG16 gene followed by connecting it to the proEM vector by T4 DNA ligase. The resulting proEM-bPAG16 recombinant vector was transfected into HEK293 cells. The expressed gene was purified by Ni2+ affinity chromatography and verified by SDS-PAGE and western blotting.   Result   With the optimization, the codon adaptation index (CAI) of the bPAG16 gene rose from 0.77 to 0.96 and the GC content from 48% to 58%. After enzymatic digestion, fragments of approximately 1 179 bp and 4 369 bp were obtained from the proEM-bPAG16 expression vector as expected. The nucleotide sequence of the inserted bPAG16 in the recombinant plasmid was same as that of the optimized gene and absent of any mutated amino acids. SDS-PAGE and western blotting showed that the 48kDa recombinant bPAG16 fusion protein was correctly expressed in the HEK293 cells with a purity greater than 90%.   Conclusion   The successfully executed codon optimization and synthesis of bPAG16 gene provided the base material for further study on early diagnosis of pregnancy in dairy cows.
Crop Science
Response and Tolerance Threshold of Bainong Aikang-58 Wheat Seedlings toward Na2SO4 Stress
LIU Duo, BAI Shuang, QI Xuebin, NING Dongfeng, LIANG Zhijie, GUO Wei, LI Ping
2021, 36(1): 9-16.   doi: 10.19303/j.issn.1008-0384.2021.01.002
[Abstract](10) [FullText HTML](7) [PDF 923KB](2)
Abstract:
  Objective  Bainong Aikang-58 is one of the major wheat cultivars in China. To investigate the response mechanism of Bainon Aikang-58 seedling to Na2SO4 stress was discussed and to define its tolerance threshold to Na2SO4 stress was determined so as to provide basic data support for further research and scientific basis and and theoretical guidance for agricultural production practice.   Method  Effects of Na2SO4 stress at 0, 20, 40, 60, 80, and 100 mmol·L−1 on the growth and physiology of the wheat seedlings were studied in a hydroponic experimentation.   Result   As the Na2SO4 concentration in the medium increased, the plant height, fresh weight, and biomass of the seedlings 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 membrane permeability and content of MDA in the synplasmic membrane increase with increasing of Na2SO4 concentration to maximize 3.2 and 4.94 times of the control. The chlorophyll content in the leaves were 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 Whereas, the proline content kept increasing to reach a maximum that was 8.51 times of control.   Conclusion  The tolerance threshold of Bainong Aikang-58 wheat seedling toward Na2SO4 stress was determined to be 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
2021, 36(1): 17-23.   doi: 10.19303/j.issn.1008-0384.2021.01.003
[Abstract](24) [FullText HTML](7) [PDF 1112KB](2)
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.
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
2021, 36(1): 24-35.   doi: 10.19303/j.issn.1008-0384.2021.01.004
[Abstract](130) [FullText HTML](58) [PDF 1252KB](8)
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 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.
Plant Protection
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
2021, 36(1): 36-40.   doi: 10.19303/j.issn.1008-0384.2021.01.005
[Abstract](118) [FullText HTML](69) [PDF 753KB](11)
Abstract:
  Objective  The rice cultivar Gufeng B confers strong, broad-spectrum, durable resistance against various rice blast isolates. The present study was aim to identify and map the blast resistance gene(s) in Gufeng B.  Method   The F1 and F2 population were obtained by crossing Gufeng B and Nipponbare, and the genetic model of blast resistance was analyzed after inoculating with 7 strains of Magnaporthe grisea. Subsequently, F2 population was used to construct a resistant pool and a sensitive pool respectively, and to map the associated loci via the method of bulked segregation analysis.   Result   Gufeng B exhibited high resistance to all of the tested strains, such as KJ201, RB22, CHNOS, RB6, 2Y838-1, 501-3 and IR16-1, suggesting that Gufeng B may carry the broad-spectrum and high resistance genes. The F1 progenies from the cross between Gufeng B and Nipponbare conferred resistance against the strains 501-3 and IR16-1, and the segregation ratio of resistance and susceptibility among F2 progenies does not fit 3:1, assuming that the resistance against the strains 501-3 and IR16-1 were controlled by multiple locus in Gufeng B. Whole genome re-sequencing of the two parental lines Gufeng B and Nipponbare identified 1,756,964 SNPs. Calculation results of △SNP-index showed that there were two candidate loci conferring resistance to rice blast disease, which were located at Chr.6: 10,082-11,397Kb, corresponding to the Pi2/9 locus, and Chr.11: 120-266Kb. 4006 SNPs and 623 InDels markers were searched within the interval of Chromosome 6, 752 SNPs and 195 InDels within the corresponding region of Chromosome 11, respectively.  Conclusion  The resistance of Gufeng B to 501-3 strain may be controlled by two resistance genes on chromosomes 6 and 11. Our results laid the foundation for finely mapping and cloning the resistance genes in Gufeng B, and provided marker resources for molecular marker-assisted selection.
Assessment on Anthracnose-resistance of 590 Soybean Cultivars in China
SHI Niuniu, DU Yixin, HE Yanqin, RUAN Hongchun, TENG Zhenyong, GAN Lin, LIAN Jinfan, YANG Zhonglu, CHEN Furu
2021, 36(1): 41-52.   doi: 10.19303/j.issn.1008-0384.2021.01.006
[Abstract](24) [FullText HTML](3) [PDF 660KB](5)
Abstract:
  Objective  Anthracnose-resistance of varieties of soybean in China was assessed and classified to facilitate the breeding of resistant cultivars.   Method  From 2011 to 2019, 590 varieties of domestic soybean cultivars were collected and challenged by an artificial inoculation of Colletotrichum truncatum to determine their degrees of resistance to anthracnose.   Result   The test showed 70 of the varieties belonged to the resistant (R), 143 to the medium resistant (MR), 219 to the medium susceptible (MS), 148 to the susceptible (S), and 10 to the highly susceptible (HS) categories, representing 11.86%, 24.24%, 37.12%, 25.08%, and 1.69%, respectively, of the entire sampled population. The overall disease resistant rate (RR) in a particular year varied from year to year, ranging from 13.64% to 68.00%, with the highest shown in 2013, followed by 52% in 2012 and the lowest in 2016. The RRs of the spring varieties in the tropical and subtropical areas (S1), the summer varieties in the tropical and subtropical areas (S2), the spring-sowing green varieties (S3), the summer-sowing green varieties (S4), the spring soybean from Yangtze river basin (S5), the new variety from Fujian (S6), and the new varieties from other regions (S7) ranked from the lowest of S3 at 5.89% to the highest of S2 at 69.23%, while S4 being the second highest at 57.69%.   Conclusion  Out of the 590 varieties, 36.10% were only resistant or moderately resistant to anthracnose with none exhibited high resistance. And, there was no spring-sowing green soybeans for fresh consumption being substantially resistant to the disease.
Inhibitory Effects of Ailanthus altissima Alkaloids on Pathogenic Fungi
LI Yongchun, ZHAO Meirong, ZHANG Zhi
2021, 36(1): 53-58.   doi: 10.19303/j.issn.1008-0384.2021.01.007
[Abstract](15) [FullText HTML](8) [PDF 750KB](0)
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 perse appeared promising to be a new source for the development of a plant-based biocontrol agent.
Pathogens Infecting Chinese Yams Planted on Ridges in Northern Hubei
YANG Xiaolin, ZHANG Youhong, WANG Zuoqian, CHANG Xiangqian, LYU Liang, ZHANG Shu, YANG Xinsun
2021, 36(1): 59-64.   doi: 10.19303/j.issn.1008-0384.2021.01.008
[Abstract](10) [FullText HTML](2) [PDF 1115KB](5)
Abstract:
  Objective   Pathogens that cause serious diseases on Dioscorea batatas Decne grown on the mountain ridges in northern Hubei province were investigated to facilitate drafting effective disease prevention and control measures.   Method   Specimens of aboveground parts of the diseased plants were collected for pathogen isolation. The isolates were identified by colonial morphology, microstructure, and blasting sequence of ITS as well as the pathogenicity determination with tissue separation of phytopathology and ITS-PCR.   Result   The isolated pathogenic fungi included Alternaria alternate, Colletotrichum fructicola, C. gloeosporiaides, and Lasiodiplodia pseudotheobromae.   Conclusion   The pathogens causing the diseases, such as anthracnose, that can lead to withering or death of Chinese yams at the region were highly diversified and complex.
Optimized Application of Fe3O4 Nanoparticles for Pesticide Detection in Tea
LI Jie, ZHANG Feng, HUANG Jingjing, DING Liping, YANG Fang, YAO Qinghua
2021, 36(1): 65-70.   doi: 10.19303/j.issn.1008-0384.2021.01.009
[Abstract](23) [FullText HTML](9) [PDF 870KB](5)
Abstract:
  Objective  A newly modified analytical method for detecting pesticide residues in tea was developed to improve test efficiency and reduce cost.   Method   Fe3O4 nanoparticles were used to replace the conventional pigment-removing material prior to measurements on pesticide residues, including carbendazim, pyrimethanil, tricyclazol, imidachloprid, acetamiprid, methomyl, and thiamethoxam, in tea by liquid chromatography-tandem mass spectrometry. The methodology was evaluated according to its applicability and validated in a spiked residue test.   Result  The recovery rates on the 7 pesticides under the modified method using 300 mg Fe3O4 nanoparticles for pigment removal pretreatment were higher than graphitizing with carbon black (GCB). The pigment-removed 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-1 ammonium acetate as Phase A and acetonitrile as Phase B for the mobile phase prior to the tandem mass spectrometric analysis under the multiple reaction monitoring mode with external standards. A linearity on the measurements by the new method was achieved within the pesticide concentrations of 0-50 µg·L-1 showing correlation coefficients (R2) greater than 0.995 for all 7 pesticides. On 5, 10, and 50 ng·g-1 spiked pesticide samples, average recovery rates of the new method ranged from 71.6 % to 107.7 % with relative standard deviations of 3.95%-13.62 %. The limits of detection (LOD) on the pesticides by the method were between 0.15 µg·kg-1 and 0.60 µg·kg-1, and the limits of quantitation (LOQ) between 0.5 µg·kg-1 and 2.0 µg·kg-1.   Conclusion  The newly developed pesticide detection method was stable, repeatable, and accurate for the designed purpose.
Resources and Environmental Science
Effect of peanut shell replacing peat soil on cultivation substrate properties and Anoectochilus roxburghii growth and quality
ZHU Qiaolian, CHI Bingjie, TONG Chenxiao, ZHANG Hongxue, HU Kun, GUO Liming, ZHOU Biqing, MAO Yanling
2021, 36(1): 71-77.   doi: 10.19303/j.issn.1008-0384.2021.01.010
[Abstract](41) [FullText HTML](15) [PDF 627KB](9)
Abstract:
  Objective  Effects of peanut shell replacing peat soil as the cultivation substrate on the substrate properties and the growth quality of Anoectochilus roxburghii were studied to obtain the optimal substrate ratio for Anoectochilus roxburghii cultivation.  Method  Five ratios of peanut shell to peat soil, including 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) and N33 (33% peat soil + 67% peanut shell), were setted to study the physical and chemical properties of the soil and the growth and quality of Anoectochilus roxburghii by pot experiment.  Result  (1) The replacement of peat soil by peanut shell improved the aeration, water holding capacity and fertility of cultivation substrate. Compared with control, the bulk density of 33% peat soil + 67% peanut shell treatment reduced by 53.84%, maximum water holding capacity and non-capillary porosity increased by 97.14% and 148.35%, and total nitrogen, available phosphorus, and vailable potassium content increased by 7.85%, 67.76%, 1830.53%, resectively, with significant difference. (2) The growth and quality of Anoectochilus roxburghiithe was the best under 33% peat soil + 67% peanut shell treatmentm, the plant height, root length, leaf length, and leaf width of which were significantly better than those of the control group; and the contents of total flavonoids, polysaccharides, free amino acids, and total phenols were significantly increased by 41.57%, 30.59%, 45.08%, and 53.16%, respectively, with significant differences.   Conclusion  33% peat soil + 67% peanut shell was the optimal ratio as substract for Anoectochilus roxburghiithe by increasing the soil fertility and improving the growth quality.
Carbon Footprint-based Temporal and Spatial Analysis on Carbon Sources/Sinks at Farmlands in Anhui Province
Zhang Jing, Fang Yu, Wei Jinda, Lin Jiahao, CHEN Peibin, Zhu Chaozhi
2021, 36(1): 78-90.   doi: 10.19303/j.issn.1008-0384.2021.01.011
[Abstract](1) [FullText HTML](0) [PDF 1173KB](1)
Abstract:
  Objective  To ensure a low-carbon agriculture for Anhui province, temporal and spatial variations on the carbon sources/sinks at the farmlands were analyzed based on the carbon footprints at selected localities.   Method   Statistical data on the related agricultural inputs and crop biomass in the province from 2008 to 2016 were applied to calculate the carbon sources/sinks and footprints using the emission coefficient method. ArcGIS and Geoda software helped visualize and auto-correlate the spatial and temporal characteristics of the selected 16 cities in 9 years.   Results   The emission intensity at the two major contributing municipalities, Huangshan and Suzhou, showed a trend of "high-in-the-north/south, low-in-the-central, and balanced-on-east/west", whereas, the absorption a mode of "heavy-on-both-ends and light-in-the-center" at Fuyang and Bengbu, the major contributing cities. The carbon emission of the ecosystem was declining during the study period, but the absorption irregular. The varied land uses over the years might contribute to the results.   Conclusion   The carbon emission intensity and absorption in the province varied with characteristic patterns. In general, the carbon footprints in the area were below the ecological tolerance capacity indicating rooms for spatial autocorrelation. Such surplus, thus, would allow the surrounding prefectures and cities a better situation in dealing with the environmental carbonization. Based on the finding, it appeared that the Action Plan for Implementing Green Development could be adequately carried out in Anhui with a scientifically planned low-carbon emission operation taking local conditions into consideration. The desirable land use and agricultural practices could be well accomplished without jeopardizing the target ecological improvement for the province.
Food Science
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
2021, 36(1): 91-103.   doi: 10.19303/j.issn.1008-0384.2021.01.012
[Abstract](23) [FullText HTML](13) [PDF 1758KB](2)
Abstract:
  Objective  Bacteriostatic property of Lactobacillus plantarum LV02 bacteriocin produced on an optimized fermentation medium was determined.   Method  Based on a single-factor design, the PB and the steepest ascent experiments were conducted to locate the center point of the step length and direction of the influencing factors for a CCD test. Subsequently, effects of different media on the bacteriostatic property and OD600 of the cultured L. plantarum LV02 were evaluated for formulation optimization. The Oxford cup method was employed to determine the bacteriostatic capacity and stability under application conditions of the resulting LV02 bacteriocin produced on the optimized medium.   Result  The optimized formula for the LV02 fermentation medium in 1 L of water constituted 34.07 g of glucose, 18.12 g of yeast extract, 2 g of dipotassium hydrogen phosphate, 0.16 g of manganese sulfate, 5 g of sodium acetate, 0.20 g of magnesium sulfate, 1 g of ammonium citrate, 1 mL of Tween 80, and 50 mL of carrot juice. On the optimized medium, LV02 grew to yield a 12% increase on OD600 and bacteriocin with a 26% increase on the diameter of inhibition zone against Escherichia coli YS. For the crude LV02 bacteriocin extraction, 80% saturation concentration of ammonium sulfate was used. The antibacterial LV02 bacteriocin was stable under 100 ℃ for 120 m and pH 3.0~7.5.   Conclusion  L. plantarum LV02 cultured on the optimized fermentation medium was bacteriostatic against E. coli YS. The fermentation produced bacteriocin with desirable stabilities to heat, acid, and slight alkaline condition and was considered promising for the development of an antibacterial agent.
Optimized Preparation of Spray-dried Anthocyanins Microcapsules
ZHAO Yanqiao, WANG Yue, MENG Xiangyu, LI Yukun
2021, 36(1): 104-114.   doi: 10.19303/j.issn.1008-0384.2021.01.013
[Abstract](0) [FullText HTML](0) [PDF 1558KB](0)
Abstract:
  Objective  TPreparation of an spray-dried encapsulated anthocyanins from Hibiscus sabdariffa was optimized.   Method   Single factor experiment and response surface method were employed for the process optimization. Structure of the microcapsules was characterized with scanning electron microscopy (SEM). Product stability under exposure of light, heat, and metal ions was determined.   Results   The optimum encapsulation used 7% total solids that consisted of 70% maltodextrin, 3% gum Arabic, and 7% H. sabdariffa anthocyanin, and the dehydration applied a spray-drying inlet-air temperature at 143 ℃. The resulting encapsulation rate reached 97.11%. Observed under SEM, the spherical microcapsules were smooth on the surface with no cracks or voids. Exposed to temperature or light, the microcapsules were significantly more stable than the purified anthocyanins. And, the product stability in the presence of Cu2+ or Zn2+ was also significantly improved by the process. A maximum shelf life of the product could be achieved by storing it at 4 ℃ in darkness and avoiding direct contact with Cu2+ or Zn2+.   Conclusion   H. sabdariffa anthocyanin microcapsules with a significantly improved shelf life were successfully prepared by the optimized microencapsulation and spray-drying.
Review
Research Progress on Genetics 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
2021, 36(1): 115-123.   doi: 10.19303/j.issn.1008-0384.2021.01.014
[Abstract](35) [FullText HTML](2) [PDF 699KB](8)
Abstract:
Variants of anthocyanins content in grain hull gives brown rice the appearance of varied colors such as black, purple, red, green, yellow, etc. These varieties of rice are, therefore, categorically termed as “colored rice”. The beneficial and functional components in colored rice include amino acids, functional lipids, dietary fiber, vitamins, minerals, anthocyanins, phenolic compounds and γ-oryzanol. Such substances are either nutritionally or medicinally advantageous to the human well-being and valued by the consumers making colored rice highly popular on the market. For instance, in comparison to research activities on the regular varieties, programs specifically targeting colored rice are relatively few and commercial products emphasizing the functionality sparse. This article reviews the status on the germplasm resource, evaluation, utilization, functional ingredients, characteristics, genetics, breeding and application of colored rice. Obstacles hinder the industrial development are analyzed. Hence, in the attempt to expand the utilization and meet the market demand of colored rice, systematic and effective studies on the germplasm genetics for advancements in breeding and the rice chemistry for new product development would be indispensable.
Research Progress on HD-ZIP Transcription Factors in Response to Pathogenic or Abiotic Stresses
LIU Jiao, SHUAI Peng
2021, 36(1): 124-134.   doi: 10.19303/j.issn.1008-0384.2021.01.015
[Abstract](208) [FullText HTML](107) [PDF 954KB](7)
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.
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