Loading...

Table of Content

    26 September 2021, Volume 37 Issue 9
    Silage Microorganisms: An Eternal Topic in the Feed Fermentation Process
    YANG Fu-yu
    2021, 37(9):  1-2. 
    Asbtract ( 353 )   HTML ( 31)   PDF (1015KB) ( 747 )  
    References | Related Articles | Metrics
    Research Progress in the Distribution of Lactic Acid Bacteria on the Surface of Plants
    TIAN Jing, ZHANG Jian-guo
    2021, 37(9):  3-10.  doi:10.13560/j.cnki.biotech.bull.1985.2021-0740
    Asbtract ( 537 )   HTML ( 28)   PDF (1083KB) ( 707 )  
    References | Related Articles | Metrics

    The species and number of lactic acid bacteria(LAB)adhering to the surface of forage crops are the key factors that determine the fermentation quality of silage,and might also affect the growth of forage crops. Studies have found that LAB on the surface of forage crops are affected by many factors. This paper discussed the effects and mechanism of plant characteristics(leaf surface structure,nutrients,chemical composition,etc.),environments(temperature,humidity,rainfall,oxygen,photoperiod,etc.)and field managements(fertilization and mowing)on the distribution of epiphytic LAB,in order to well understand the situation of LAB on the surface of plants. Thus,we may increase the types and quantities of excellent LAB on the surface of plants through planting management and harvesting techniques,which will provide scientific bases for the production of high-quality silage and the healthy growth of plants.

    Research Advances in Silage Microbial Communities and Functions
    CHEN Meng-yan, BAI Jie, KE Wen-can, XU Dong-mei, AI Lin, GUO Xu-sheng
    2021, 37(9):  11-23.  doi:10.13560/j.cnki.biotech.bull.1985.2021-0926
    Asbtract ( 616 )   HTML ( 27)   PDF (1133KB) ( 852 )  
    Figures and Tables | References | Related Articles | Metrics

    Microbial community is a key factor affecting the fermentation quality of silage. Both microorganisms attached to the surface of raw materials and exogenous microorganisms play an important role in the microbial community succession during the ensiling process. Understanding the microbial community,function and fermentation metabolism regulation network of silage are of great significance for in-depth analysis of the biological process of silage fermentation,and also can provide a strong scientific basis for the fermentation and modulation of safe and high-quality silage. However,the traditional detection methods are of low accuracy and cannot deeply analyze the microbial community structure in silage. Therefore,how to comprehensively and accurately use molecular biology techniques to detect the microbial community structure and species in silage will be of great significance for breaking through the research of micro-ecological regulation of silage fermentation. This review depicted the effects of epiphytic microorganisms and growth environment,characterization of silage materials,and lactic acid bacteria inoculants on the microbial structure of silage,as well as summarized the current situation of PICRUSt method applied in silage microbial function,aiming to provide some suggestions for characterizing microbial dynamics and function of silage and to present theoretical foundation for revealing micro-ecology process during the ensiling process.

    Research Progress in the Effects of Additives to Silage on Microbial Diversity
    XIN Ya-fen, CHEN Chen, ZENG Tai-ru, DU Zhao-chang, NI Hao-ran, ZHONG Yi-hao, TAN Xiao-ping, YAN Yan-hong
    2021, 37(9):  24-30.  doi:10.13560/j.cnki.biotech.bull.1985.2021-0818
    Asbtract ( 577 )   HTML ( 24)   PDF (1097KB) ( 769 )  
    References | Related Articles | Metrics

    Ensiling of forage is a very complex process of microbial activities and biochemical changes. The quality of silage crops depends on the microbial community during the early phase of fermentation and its succession in the fermentation process. Additives play an important role to change the microbial diversity and to enhance the silage quality. They improve the fermentation quality and aerobic stability mainly by promoting the rapid fermentation of lactic acid bacteria and other dominant bacteria,consequently reducing the pH,and creating an acidic environment to inhibit the growth and reproduction of spoilage microorganisms. The current paper is aimed to review the effects of lactic acid bacteria,chemical,enzymatic and nutritional additives on the microbial community structure of silage,aiming to provide best references for studying the mechanism of microbial actions during the fermentation process of silage,and directional research and development of silage additives in the future.

    Research Progress in the Succession of Microbial Communities in Total Mixed Ration Silage
    JIANG Di, XU Chun-cheng
    2021, 37(9):  31-38.  doi:10.13560/j.cnki.biotech.bull.1985.2021-0832
    Asbtract ( 459 )   HTML ( 10)   PDF (1084KB) ( 608 )  
    Figures and Tables | References | Related Articles | Metrics

    Total mixed ratio(TMR)silage is a kind of nutritional balanced rations fermented by lactic acid bacteria according to a scientific diet formula based on the nutrition needs at different growth stages of livestock and the nutritional value of feed materials. With the rapid development of animal husbandry in China,the demand for high-quality feed is increasing. In order to reduce the excessive dependence on feed grain,the development and utilization of unconventional feed resources is particularly important. The expansion and popularization of modulating and processing technology for fermenting TMR silage is beneficial to the efficient utilization of by-products to solve the shortage of feed resources that restrict the stable development of animal husbandry. And the microbial communities of TMR silage play an important role in the fermentation quality and aerobic stability. Therefore,this article reviews the application of TMR silage technology,microbial factors affecting aerobic stability,and the diversity of microbial communities in TMR silage,and discusses the significance and existing problems in TMR silage technology. It is of great significance to realize the production potential,to promote this feeding technology to efficiently apply it to production practice. And this would provide new ideas for improving the utilization efficiency of unconventional feed resources and solving the problem of shortage in feed resources.

    Excellent Lactic Acid Bacteria for Silage and Their Application
    XU Jin-yi, NA Bin-bin, LIU Shun, CHEN Chao, SUN Hong, ZHENG Yu-long
    2021, 37(9):  39-47.  doi:10.13560/j.cnki.biotech.bull.1985.2021-0806
    Asbtract ( 585 )   HTML ( 15)   PDF (1064KB) ( 618 )  
    Figures and Tables | References | Related Articles | Metrics

    Silage is an important storage method which can effectively preserve feed nutrients. Silage fermentation is a very complex microbial activity system,whether the fermentation process can be effectively controlled is the key to the success of silage. By adding excellent lactic acid bacteria in silage,the microbial flora in silage can be reasonably adjusted to control the fermentation process and improve the quality of silage,and thus to screen excellent lactic acid bacteria for promoting the rapid development of silage industry. Therefore,based on the researches on the screening and utilization of excellent silage lactic acid bacteria at home and abroad,this paper summarized the types,action mechanism,separation and screening,and application of excellent lactic acid bacteria in silage,and put forward suggestions and prospects for its future development,aiming to provide some reference for the subsequent screening of excellent lactic acid bacteria and the development of high-quality lactic acid bacteria preparation.

    Research Progress in Woody Forage Silage
    ZHANG Ying-chao, YIN Shou-liang, WANG Yi-wei, WANG Xue-kai, YANG Fu-yu
    2021, 37(9):  48-57.  doi:10.13560/j.cnki.biotech.bull.1985.2021-0913
    Asbtract ( 423 )   HTML ( 17)   PDF (1120KB) ( 694 )  
    References | Related Articles | Metrics

    With the rapid development of animal husbandry in China,the demand for high-quality roughage is growingly increasing,and the supply of forage is more and more scarce. Therefore,it is important to exploit new forage which can alleviate the shortage of traditional forage. Woody forage is a new type of forage that is characterized by abundant resources,high yield and high crude protein content. Reasonable development and utilization of woody forage may alleviate the shortage of forage resources. Silage is an important storage method of woody forage,which can preserve forage nutrients with high quality. In recent years,more and more researches have being focused on woody forage silages. This paper summarized the nutritional characteristics,quality of natural ensiling,improvement methods of woody forage silages,application of woody forage silages in livestock production and brought out suggestions for future research,aiming at providing reference for the utilization and development of woody forage silages.

    Isolation,Identification of Lactic Acid Degrading Bacteria in Alfalfa Silage and Their Degradation Characterization
    CUI Xin-yu, LI Rong-rong, CAI Rui, WANG Yan, ZHENG Meng-hu, XU Chun-cheng
    2021, 37(9):  58-67.  doi:10.13560/j.cnki.biotech.bull.1985.2021-0813
    Asbtract ( 399 )   HTML ( 13)   PDF (3390KB) ( 468 )  
    Figures and Tables | References | Related Articles | Metrics

    In order to solve the lactic acid degradation problem during alfalfa silage,three varieties of alfalfa were selected for silage,and sodium lactate was used as the sole carbon source to isolate microorganisms capable of degrading lactic acid. The morphological observation,physiological and biochemical characteristics and 16S rDNA sequence analysis were employed to identify the strains,and the UV spectrophotometry and gas chromatography were used to study degradation rate and characteristics of the strains. A total of 75 strains of lactic acid degrading bacteria were obtained,among which RSM9,RSF15,RSF2 and RSH16 with high degradation rate were selected. They were identified as Hafnia sp.,Proteus sp.,Obesumbacterium sp. and Citrobacter sp. The degradation rates of lactic acid reached 44.64%,33.86%,30.64% and 33.35% after 120 h of anaerobic culture at 30℃ and pH 6.2,respectively. The acid metabolites of the 4 strains were mainly acetic acid and propionic acid,and 4 strains were effectively inhibited at pH<5.0. NAD-independent lactate dehydrogenases and lactate oxidase were detected in all strains except for RSH16. Four strains of lactic acid degrading bacteria are screened from the process of alfalfa silage,as well as,their metabolism of lactic acid can increase the pH of the system and cause the decline of alfalfa silage quality.

    Effects of Addition Amount of Molasses on the Fermentation Quality and Microbial Diversity of Hybrid Broussonetia papyrifera L. Vent Silage
    JIANG Fu-gui, CHENG Hai-jian, WEI Chen, ZHANG Zhao-kun, SU Wen-zheng, SHI Guang, SONG En-liang
    2021, 37(9):  68-76.  doi:10.13560/j.cnki.biotech.bull.1985.2021-1104
    Asbtract ( 386 )   HTML ( 11)   PDF (3781KB) ( 357 )  
    Figures and Tables | References | Related Articles | Metrics

    This experiment was conducted to investigate the effects of addition amount of molasses on nutritional value,fermentation quality and microbial diversity of hybrid Broussonetia papyrifera L. Vent silage. The four treatments were control group(without additive),M5 group(5 g/kg molasses),M10 group(10 g/kg molasses)and M20 group(20 g/kg molasses). Each group had four replicates and ensiled for 60 d. As results,with an increase of molasses addition amount,the contents of dry matter and crude protein of hybrid B. papyrifera L.Vent silages linearly increased(P < 0.05),while the contents of neutral detergent fiber and acid detergent fiber linearly decreased(P < 0.05). With an increase of molasses addition amount,the lactic acid content of hybrid B. papyrifera L.Vent silages linearly increased(P = 0.002),while pH,butyric acid content and NH3-N/TN linearly decreased(P < 0.05). At the phylum level,the dominant phylum in hybrid B. papyrifera L.Vent silages were Firmicutes,followed by Proteobacteria. At the genus level,the proportion of Lactobacillus significantly increased in the M10 and M20 group than the CK and M5 group(P = 0.005),while the proportion of Weissella significantly decreased(P = 0.003). The proportions of Clostridium_sensu_stricto_12 and Enterobacter in the M5,M10 and M20 groups were lower than those in the CK group(P < 0.05). In conclusion,the addition of molasses improved the nutritional value and fermentation quality of hybrid B. papyrifera L. Vent silage,increased the proportion of Lactobacillus,and reduced the proportion of Clostridium_sensu_stricto_12 and Enterobacter. The hybrid B. papyrifera L. Vent silage added with 20 g/kg molasses had the best nutritional value and fermentation quality.

    Effects of Lactobacillus paracasei on the Quality and Bacterial Diversity of Silage Alfalfa After Aerobic Exposure
    WANG Qi, WU Zhi-xuan, CHEN Zhong-ling, WU Bai-yi-la, HU Zong-fu, NIU Hua-xin
    2021, 37(9):  77-85.  doi:10.13560/j.cnki.biotech.bull.1985.2021-0717
    Asbtract ( 384 )   HTML ( 13)   PDF (4602KB) ( 348 )  
    Figures and Tables | References | Related Articles | Metrics

    16S rRNA high-throughput sequencing technology was used to detect the dynamic variations of bacterial community in the silage alfalfa after aerobic exposure,aiming to provide a basis for the dynamic changes of bacterial diversity in the aerobic exposure of silage alfalfa and the inoculation of lactic acid bacteria to prevent aerobic deterioration. Alfalfa silage without inoculation(CON)and inoculated with Lactobacillus paracasei(LCP)were fermented for 56 d,and the dynamic changes of fermentation quality and bacterial diversity of the silage at 0,7,and 14 d after aerobic exposure. The results showed that aerobic exposure resulted in the increase of the pH of uninoculated silage alfalfa and the decrease of the lactic acid content. Compared with CON,inoculation of L. paracasei had a smaller increase in pH but higher lactic acid content during aerobic exposure(P<0.05). CON was mainly composed of Lactobacillus,Enterobacter,and Enterococcus,among which the abundance of Enterobacter and Enterococcus gradually decreased;the LCP group was mainly dominated by Lactobacillus,and its abundance gradually decreased. In addition,abundance of Acetobacter appeared after 14 d of aerobic exposure. Correlation analysis revealed that Lactobacillus was negatively correlated with pH(P<0.05). Enterobacter,Enterococcus,Wierella,Cedecea,Sporolactobacillus were positively correlated with pH. Acetobacter was negatively correlated with lactic acid and acetic acid(P<0.05). In summary,this study found that Acetobacter was abundant in the silage alfalfa exposed to long-term aerobic exposure,reducing the content of lactic acid and acetic acid. Inoculation of L. paracasei may increase the abundance of Lactobacillus and reduce the abundance of Acetobacter in long-term(14 d)aerobic exposure. Therefore,inoculation of silage alfalfa with L. paracasei can improve the quality of short-term aerobic exposure silage,enhance its aerobic stability,and slow down aerobic deterioration.

    Effects of Microbial Inoculants on the Fermentation Quality and Microbial Community Diversity of Alfalfa Silage
    MAO Ting, NIU Yong-yan, ZHENG Qun, YANG Tao, MU Yong-song, ZHU Ying, JI Bin, WANG Zhi-ye
    2021, 37(9):  86-94.  doi:10.13560/j.cnki.biotech.bull.1985.2021-0831
    Asbtract ( 410 )   HTML ( 12)   PDF (2892KB) ( 381 )  
    Figures and Tables | References | Related Articles | Metrics

    In order to develop microbial inoculants suitable for alfalfa silage,the relationship between the microbial community of silage and the quality of alfalfa silage was investigated. The experiment was conducted to isolate and screen lactic acid strains with high acid production capacity from the alfalfa fermented green juice. The 16S rDNA gene sequence homology analysis was used to identify and then to prepare silage bacteria GSSW in compound. Silage samples were divided into 4 groups,adding with fermented green juice(aFGJ),sila-max silage(YB),GSSW bacteria,and no addition as control(CK). A high throughput sequencing technique was used to analyze the microbial community diversity and detect physicochemical fermentation indexes. The results showed that MXLZ-1,MXLZ-2 and MXLZ-4 isolated from the fermented green juice were identified as Pediococcus pentosaceus,Lactobacillus plantarum and Pediococcus parvulus. Compared with CK group,dry matter and crude protein in the other 3 groups significantly increased(P<0.05);neutral detergent fiber,acid detergent fiber and lignin significantly decreased(P<0.05);pH and ammonia nitrogen/total nitrogen significantly decreased(P<0.05),lactic acid significantly increased(P<0.05). Lactobacillis and Pediococcus were the genera with high relative abundance of silage samples in the 4 groups. The abundance of Lactobacillis was higher than that of Pediococcus group with microbial inoculants,while that of CK was the opposite. Lactobacillus plantarum,Pediococcus pentosaceus,Lactobacillus brevis,and Enterococcus mundti of silage samples in the 4 groups were significantly different. Adding three kinds of microbial inoculants increased the abundance of L. plantarum and decreased the abundance of P. pentosaceus and E. mundtii. Adding silage inoculants increased the number of beneficial bacteria,reduced the number of harmful bacteria,and improved the fermentation quality of alfalfa silage. The GSSW can be used in alfalfa silage.

    Study on the Detoxification Characteristics of Antifungal Lactic Acid Bacteria and the Application of Silage
    ZHAO Ya-ru, XU Qing-fang, GAO Wen-jun, GUO Gang, CHEN Lei, YU Zhu
    2021, 37(9):  95-105.  doi:10.13560/j.cnki.biotech.bull.1985.2021-0897
    Asbtract ( 382 )   HTML ( 13)   PDF (4678KB) ( 412 )  
    Figures and Tables | References | Related Articles | Metrics

    This study aims to add Lactobacillus plantarum to inhibit the proliferation of silage mold and degrade toxins in order to improve feed safety. HPLC was adapted to determine the antibacterial components in the cell-free supernatant(CFS)of L. plantarum and to evaluate the adsorption efficiency of AFB1. After that,L. plantarum and Aspergillus flavus were added to the whole corn(Zea mays L.)silage,the silage at 0,24 and 48 d of fermentation was evaluated by sensory,and the fermentation quality,nutritional value,microorganism,aerobic stability and feed mildew condition were determined and analyzed. The results showed that there were organic acids in CFS,and the adsorption rate of AFB1 by L. plantarum reached 59.40%. The sensory quality,microbial composition,fermentation performance,nutrient composition,feed value,CNCPS,aerobic stability,and feed mildew of the silage treated with L. plantarum were significantly better than those of the control group and the A. flavus treatment group(P <0.05),no toxin was detected in each treatment group. L. plantarum destroyed the mold structure,thereby inhibiting its growth,and had the ability to adsorb toxins,which effectively improved the fermentation quality and nutritional components of the whole plant corn silage,and prevented the feed from being infected by molds that leads to reduced safety.

    Molecular Cloning,Expression and VIGS Construction of a Small GTP-binding Protein Gene GhROP3 in Gossypium hirsutum
    HU Zi-yao, DAI Pei-hong, LIU Chao, Madina Mulati, WANG Qian, Wugalihan Abuduwili, ZHAO Yi, SUN Ling, XU Shi-jia, LI Yue
    2021, 37(9):  106-113.  doi:10.13560/j.cnki.biotech.bull.1985.2021-0287
    Asbtract ( 428 )   HTML ( 15)   PDF (3180KB) ( 384 )  
    Figures and Tables | References | Related Articles | Metrics

    This study is to dissect Gossypium hirsutu’s small GTP-binding protein gene GhROP3 expression patterns responding to different abiotic stresses,thus providing a foundation for stress-related genes cloning and elucidating the molecular mechanism of G. hirsutu stress-resistance. Homologous approach was used to clone the GhROP3,and bioinformatics methods were employed to analyze the physical and chemical properties of the gene. qRT-PCR was to investigate the tissue-specific expressions and patterns of GhROP3 under various induction conditions. The VIGS silencing vector of the gene was constructed and transformed into cotton by Agrobacterium-mediated method,and the silencing efficiency was detected by qRT-PCR. As results,the GhROP3 was cloned from the cDNA of G. hirsutu,it contained a 591 bp open reading frame,encoded a basic and hydrophilic protein containing 196 amino acid residues,the relative molecular mass was 21.75 kD. qRT-PCR analysis showed that GhROP3 expressed in the roots,stems,leaves,cotyledons and hypocotyl of G. hirsutu seedlings,and especially at a relatively higher level in stems. GhROP3 expression responded differentially to high salinity,drought,low temperature and Verticillium wilt. The VIGS silencing vector of the gene was constructed and transformed into cotton. The silenced GhROP3 in the leaves and roots of cotton was detected by qRT-PCR,which proved that the VIGS silencing vector was successfully constructed and worked normally in plants. The results suggested that GhROP3 may play important roles in the adversity stress adaption of G. hirsutu plant.

    Expression Analysis and Functional Verification of the HaACO1 Gene in Sunflower
    SUN Rui-fen, ZHANG Yan-fang, NIU Su-qing, GUO Shu-chun, LI Su-ping, YU Hai-feng, NIE Hui, MOU Ying-nan
    2021, 37(9):  114-124.  doi:10.13560/j.cnki.biotech.bull.1985.2020-1482
    Asbtract ( 513 )   HTML ( 18)   PDF (4508KB) ( 438 )  
    Figures and Tables | References | Related Articles | Metrics

    To exploit genes encoding ACC oxidase in sunflower more profitably,we studied one ACC oxidase gene HaACO1(GenBank accession number:KP966508)cloned from previous salt-induced sunflower plants. The expression patterns of HaACO1 under different stress types were investigated. Additionally,the overexpression of this gene in tobacco was carried out. The results showed that the expression of HaACO1 in sunflower was induced by pathogens,mechanical damage,low temperature,NaCl and salicylic acid stress,and expression patterns were distinct under these types of stress. HaACO1 expressed in the roots,hypocotyls and leaves of sunflower plants,with the highest expression in the leaves and the lowest expression in the roots. By taking advantage of a transient expressive vector,the subcellular localization of HaACO1-GFP was found to be largely in the cytoplasm of onion epidermal cells. A plant vector of expressing HaACO1 was constructed to analyze the overexpressing of HaACO1. The results indicated the decrease in green color of leaves of transgenic tobacco was smaller than that of the wild plants placed on differentiation medium with NaCl,while the leaf differentiation was higher than that of the wild plants. Under low temperature,drought and NaCl stresses,the relative expression of HaACO1 in the transgenic tobacco plants was higher than that of the wild plants. During NaCl stress,the contents of total soluble protein,proline and chlorophyll and the activities of peroxidase and superoxide dismutase in the transgenic tobacco plants increased;and the expression of a key gene P5CS for proline biosynthesis and three antioxidant related genes POD,MnSOD and CuZnSOD were all up-regulated. These results indicate that overexpression of HaACO1 in tobacco plants enhances the plants’ tolerance to high concentrations of NaCl,providing foundations for further understanding the molecular mechanism of salt tolerance in sunflower,and laying a foundation for using this gene in crop stress resistance improvement.

    Cloning,Expression and Bioinformatics Analyses of CpBURP from Commelina purpurea
    PENG Guo-ying, LU Shan, HUANG Chao, YANG Kun, WAN Wei, HUANG Chang-gan
    2021, 37(9):  125-131.  doi:10.13560/j.cnki.biotech.bull.1985.2020-1572
    Asbtract ( 360 )   HTML ( 18)   PDF (5044KB) ( 426 )  
    Figures and Tables | References | Related Articles | Metrics

    BURP protein plays an important role in the growth of plant and resistance to abiotic stress. Cloning a CpBURP and analyzing its role in Commelina purpurea may lay a foundation for further deeply studying the gene BURP in C. purpurea. Screening was performed based on the C. purpurea transcriptome database and a CpBURP was cloned,and then its bioinformatics and expressions were analyzed. The result showed the CpBURP of 1 383 bp encoded a protein with 460 amino acids. The hydrophilic CpBURP protein contained a BURP conserved domain and 50 phosphorylation sites,but no transmembrane structure and signal peptide region. It is predicted that CpBURP protein acted on the cytoplasm and peroxisomes. Phylogenetic tree analysis revealed that CpBURP protein and 13 plant BURP proteins were grouped into 6 subcategories,the CpBURP protein had the closest affinity to the BURP protein in Brassica napus. The results of qRT-PCR demonstrated that the expression of CpBURP in the root was higher than that in the stem and leaf. Under the Cu2+ stress,the expression of CpBURP in the roots was significantly higher than that in the control group. CpBURP may play an important role in the response of C. purpurea to Cu2+ stress.

    Cloning and Functional Identification of Monoterpene Synthase Gene NtTPS2 in Tobacco
    LIU Shao-hua, ZHAO Xi-sheng, YANG Qing, YANG Chang-qing, PAN Xu-hao, ZHANG Jian-hui, YANG Ai-guo, LI Yi-ting
    2021, 37(9):  132-141.  doi:10.13560/j.cnki.biotech.bull.1985.2020-1492
    Asbtract ( 609 )   HTML ( 17)   PDF (5107KB) ( 391 )  
    Figures and Tables | References | Related Articles | Metrics

    Terpenoids constitute the most abundant and diverse family in structure and function of the secondary metabolites of plants. They play important roles in the regulation of plant growth and development,stress responses and interactions with environmental biotic and abiotic factors. To study the biosynthesis of terpenoids in tobacco(Nicotiana tabacum),a monoterpene synthase was cloned from N. tabacum,named as NtTPS2. NtTPS2 contained an open reading frame(ORF)of 1 854 bp,encoding a protein of 617 amino acids with a N-terminal signal peptide and conserved DDxxD and NSE/DTE domains. Phylogenetic analysis revealed that NtTPS2 was in the TPS-II clade sharing high amino acid sequence identities with monoterpene synthase genes from petunia(Petunia hybrida),lemon(Citrus limon)and citrus(Citrus reticulata Blanco). NtTPS2 was located in the chloroplast in plant cells. Real-time PCR analysis showed that NtTPS2 expressed abundantly in the roots and pistils,and responded to the treatments of low temperature,high salt,drought and ABA. When NtTPS2 was co-expressed in Escherichia coli with mevalonate pathway and geranyl diphosphate synthase,the formation of monoterpene geraniol and its isomer nerol was detected. These results suggest that NtTPS2 plays a key role in the biosynthesis and metabolism of monoterpene in tobacco.

    Effects of 60Co-γ Radiation on the Seedling Rate and Plant Characteristics of Pinellia ternata Callus
    ZHAO Zhong-ying, LI Qing-miao, TIAN Meng-liang, YANG Xiao-qian, KANG Yao, QIU Yu-jie, ZHANG Qing-ling, LIU Fan
    2021, 37(9):  142-151.  doi:10.13560/j.cnki.biotech.bull.1985.2020-1541
    Asbtract ( 354 )   HTML ( 6)   PDF (6166KB) ( 563 )  
    Figures and Tables | References | Related Articles | Metrics

    The objectives are to explore the effects of different 60Co-γ-rays on the seedling rate and plant characteristics of Pinellia ternata callus and to determine the half lethal dose of radiation treating P. ternata. The P. ternate callus was irradiated with different doses of 60Co-γ-ray,and then the differentiation rate,seedling rate,mortality rate,plant growth and variation were analyzed and compared. The results showed that there was a negative correlation between the bud differentiation rate and radiation dose in the range of 0-30 Gy;the half lethal dose calculated by regression equation was 22.34 Gy;the plant height and tuber size of the partially regenerated plants after irradiation were lower than those of the control group,and the fresh weight of tuber,leaf type,location of bulbils,number of bulbils,chlorophyll content and esterase isozymes of the partially regenerated plants after irradiation presented non-directional variation. According to the morphological,physiological and biochemical indexes,13 mutant plants were selected. The mutation frequency was the highest under 25 Gy treatment,and 5 mutant plants were obtained,which was close to the half lethal dose and was the optimal radiation dose.

    Effects of Foliar Gradient Micro-fertilizer Sprayed by UAV on the Grain Mineral Elements of Different Winter Wheat Varieties
    LI Wen-zong, LI Chun-ping, LIANG Xin, WANG Run-hao, WANG Lei
    2021, 37(9):  152-160.  doi:10.13560/j.cnki.biotech.bull.1985.2020-1434
    Asbtract ( 364 )   HTML ( 9)   PDF (3222KB) ( 300 )  
    Figures and Tables | References | Related Articles | Metrics

    A two-factor split-plot design method was adopted to conduct UAV foliar spraying in 3 wheat varieties at 4 different concentrations of iron,zinc and selenium,in order to explore their influences on the contents of iron,zinc,selenium and other mineral elements in grains,so as to seek the optimal combination of foliar spraying concentration and wheat varieties. The results showed that the grains’iron,zinc and selenium contents of Zhengmai 0943 significantly increased under medium concentration treatment(2.5% FeSO4·7H2O + 2.5% ZnSO4·7H2O + 0.1% Na2SeO3). The zinc content in the grains of Bainong AK58 and Zhengyumai 958 significantly increased under different concentration treatments(T1:0.5% FeSO4·7H2O + 0.5% ZnSO4·7H2O+ 0.02% Na2SeO3,T2:2.5% FeSO4·7H2O + 2.5% ZnSO4·7H2O + 0.1% Na2SeO3,T3:5% FeSO4·7H2O + 5% ZnSO4·7H2O + 0.2% Na2SeO3),while the iron and selenium contents in the grains did not vary significantly under different concentration treatments. There were significant differences in the effects of variety,concentration and interaction on the grains’iron,zinc,and selenium content,and variety was the major factor,and followed by foliar spraying concentration. Foliar spraying of different concentrations of iron,zinc,and selenium mixed micro-fertilizers by UAV demonstrated overall little effect on the P,Ca,Mg,Cu,and Mn contents of 3 wheat varieties. Under this experimental treatment,the contents of iron,zinc and selenium in the wheat grains significantly increased to achieve the purpose of nutritional enhancement. Therefore,it is recommended to plant Bainong AK58 for iron-rich wheat,Bainong AK58 and Zhengmai 0943 for zinc-rich wheat,and Zhengmai 0943 for selenium-rich wheat.

    Biological Function of a Zn2Cys6 Transcription Factor CgAswA in Colletotrichum gloeosporioides
    LIU Sha-yu, CAO Jian, LI Meng, LIU Zhi-qiang, LI Xiao-yu
    2021, 37(9):  161-170.  doi:10.13560/j.cnki.biotech.bull.1985.2021-0047
    Asbtract ( 339 )   HTML ( 10)   PDF (4500KB) ( 238 )  
    Figures and Tables | References | Related Articles | Metrics

    Zn2Cys6-type transcription factor is a unique regulation factor in fungi and plays an important regulatory role in the growth,development and pathogenicity of plant pathogenic fungi. Currently,there are few reports on the transcription factor of Colletotrichum gloeosporioides. The gene of CgaswA was amplified by PCR and bioinformatics analysis was conducted. The CgaswA knock-out mutant was obtained by homologous recombination method,and the complementary strain was constructed on the basis of the mutant strain. Further,the biological function of CgaswA was determined by the phenotypic analyses of vegetative growth,conidia,production,appressorium formation and pathogenicity,etc. Results showed that gene CgaswA was acquired by PCR amplification,and it encoded a 713-amino acids protein. Compared with the wild-type strain,the knock-out mutant showed slow vegetative growth,low conidial yield,delayed spore germination and appressorium formation,and reduced pathogenicity. In conclusion,CgAswA is involved in the regulation of vegetative growth,conidiation,germination,appressorium formation and invasion,and pathogenicity in C. gloeosporioides.

    Effects of Glucose-xylose Co-utilization on the Synthesis of D-1,2,4-Butanetriol by Recombinant Escherichia coli
    LIU Xue-dan, YANG Meng, ZHANG Jing, ZHAO Dong-xu
    2021, 37(9):  171-179.  doi:10.13560/j.cnki.biotech.bull.1985.2021-0033
    Asbtract ( 381 )   HTML ( 17)   PDF (4186KB) ( 289 )  
    Figures and Tables | References | Related Articles | Metrics

    The purpose of this study is to investigate the effects of co-utilization of glucose and xylose on D-1,2,4-butanetriol(BT)biosynthesis by recombinant Escherichia coli,and to optimize the culture condition for BT production when glucose and xylose were used as mixed carbon source. By knocking out the genes affecting glucose phosphotransferase system(PTS),a series of BT-synthesized E. coli that could co-utilize a mixture of glucose and xylose were constructed,the shake-flask culture conditions of mtfA knockout and mlc overexpression strain MJ133k-ΔmtfA-PTMXM were optimized. The results showed that the strains with the genes of ptsG,mtfA and pgi deleted respectively used glucose and xylose simultaneously,and BT biosynthesis was improved. The culture conditions of strain MJ133k-ΔmtfA-PTMXM synthesizing BT was as follows:1.5 × LB was used as culture medium,in which 1% CaCO3 was added to maintain pH stable,culture temperature was kept at 33℃,medium volume of 60 mL was used in 250 mL flask. Twenty gram per liter xylose was added to the medium at 6 h and 5 g/L glucose was added twice at 6 h and 24 h respectively. The yield of BT reached 3.36 g/L,which was 4.15 times higher compared to that by original strain. In conclusion,co-utilization of glucose and xylose effectively improves BT production by the recombinant E. coli.

    Expression,Purification,and Crystallization of Pif1 Helicase from Bacteroides fragilis
    CAO Ru-fei, LI Ze-xuan, XU Huan, ZHANG Sha, ZHANG Min-min, DAI Feng, DUAN Xiao-lei
    2021, 37(9):  180-190.  doi:10.13560/j.cnki.biotech.bull.1985.2020-1419
    Asbtract ( 479 )   HTML ( 11)   PDF (5818KB) ( 519 )  
    Figures and Tables | References | Related Articles | Metrics

    To explore the structure and function of Pif1 helicase from Bacteroides fragilis,we obtained the monocrystal of B. f Pif1 for X-ray diffraction. The prokaryotic expression vector pET15B-SUMO-B.f Pif1 was successfully constructed,and the expression of recombinant B. fragilis Pif1 protein was induced. A series of protein purifications were performed,including Ni-NTA,SUMO-endonuclease digestion,DEAE ion-exchange column chromatography,and S200 gel filtration chromatography. The activity of the purified B. fragilis Pif1 protein was detected using the Stopped-Flow technique. Then,the crystallization by robot screening was performed with a variety of kits following by the optimization of crystal culture conditions,and the preliminary X-ray diffraction analysis was conducted. The B. fragilis Pif1 protein with high purity(>98.5%)and high concentration(17 mg/mL)was obtained. The kinetic results showed that purified protein had good Pif1-helicase activity. Crystallization experiments demonstrated that the monocrystals of B. fragilis Pif1 protein were obtained under the conditions of 0.1 mol/L Bis-Tris acetic acid(pH 8.3),0.05 mol/L sodium bicarbonate,5% glycerol,and 0.015 mol/L spermidine,and its resolution of X -ray diffraction reached 3.5 Å. It is the first time to successfully express,purify and obtain the monocrystal of B. fragilis Pif1 protein with high X-ray diffraction.

    Change of Differentially Expressed Genes and SNP Before or After Pseudomonas aeruginosa Resistance to Tachyplesin I
    HONG Jun, WEI Xia-yi, JI Bing-jie, YE Yan-xin, CHENG Tian-ci
    2021, 37(9):  191-202.  doi:10.13560/j.cnki.biotech.bull.1985.2020-1445
    Asbtract ( 414 )   HTML ( 9)   PDF (3377KB) ( 391 )  
    Figures and Tables | References | Related Articles | Metrics

    To explore the resistance mechanism of Pseudomonas aeruginosa to antibacterial peptide tachyplesin I,we employed RNA sequencing to analyze the changes of differentially expressed genes,sRNA target genes,and SNP between tachyplesin I-resistant strain and the original strain. By GO function and KEGG pathway enrichment,the results showed that differentially expressed genes were the most related to the functions of integral component of membrane,nucleotide binding,formate dehydrogenase(NAD+)activity,and there was no significant enrichment pathway. SNP of 22 differentially expressed genes changed,which was related to known genes encoding lipid A deacylase,outer membrane protein,cold shock protein,and lipopolysaccharide modification and hypothesized proteins. Further prediction and analysis uncovered 11 differentially expressed sRNAs and corresponding 863 differentially expressed target genes. The functions of these sRNA target genes were mainly related to histidine biosynthetic process,homoserine kinase activity,and 5-carboxymethyl-2-hydroxymuconate delta-isomerase activity. It is referred that the resistance mechanism of P. aeruginosa to tachyplesin I may be regulated by affecting the synthesis and metabolism of amino acids,the formation and modification of membrane proteins,and metabolism of iron ions,as well as by leading to the base mutations of few genes. At the same time,sRNA played its regulatory role by acting these related target genes expression. The SNP mutated genes and its mRNA regulations to their target genes by sRNA remain to be further verified.

    Effect of Low Crude Protein Diet on the Liver Transcriptome Sequencing of Goats
    ZHU Wen, TANG Ying-ying, SUN Xin-yang, ZHOU Ming, ZHANG Zi-jun, CHEN Xing-yong
    2021, 37(9):  203-211.  doi:10.13560/j.cnki.biotech.bull.1985.2020-1505
    Asbtract ( 527 )   HTML ( 22)   PDF (5940KB) ( 825 )  
    Figures and Tables | References | Related Articles | Metrics

    In order to further apply the low protein diet(LCP)feeding technology and investigate the regulation mechanisms of LCP on goat liver function,RNA-Seq technology was employed to obtain the transcriptome of the liver tissue of Anhui white goat fed LCP or normal diet. Go annotation and KEGG analysis were used to determine the functions of the differentially expressed genes(DEGs),and PPI was employed for the analysis of DEGs interactions. Results showed that there were 62 genes(FDR<0.05)with over two-fold expression variation of genes in the goat liver after LCP feeding,among which 49 genes were up-regulated and 13 genes were down-regulated. A series of DEGs related to protein metabolism,energy and lipid metabolism were identified. The functions of these DEGs were classified into 42 GO items,and the DEGs were mainly concentrated in binding,catalytic activity,molecular transducer activity and other functions. KEGG analysis showed that 23 metabolic pathways were significantly(P<0.05)enriched,including taste transduction,fatty acid biosynthesis,insulin signaling pathway,and so on. Three core DEGs including PPP1R3B,FASN,and IRS2 were enriched in insulin signal pathway in the PPI network. Above results indicate that LCP significantly affects the metabolism of glucose and lipid in the goat liver,increase fatty acid synthesis and lipid peroxidation,and decrease hepatic gluconeogenesis,which provides theoretical basis for the nutrition regulation of low protein diet.

    Effects of Crude Polysaccharide from Cistanche deserticola in Xinjiang on Foot-and-Mouth Disease Viral Vaccine Antibody and T cell Subgroup
    ZHANG Ai-lian, BA Xue-li, WANG Dan-yang, ZHAO Bing
    2021, 37(9):  212-218.  doi:10.13560/j.cnki.biotech.bull.1985.2020-1455
    Asbtract ( 382 )   HTML ( 7)   PDF (2925KB) ( 221 )  
    Figures and Tables | References | Related Articles | Metrics

    To screen new polysaccharide vaccine adjuvants of foot-and-mouth disease,the crude polysaccharide from Cistanche deserticola in Xinjiang(CDCP)and inactivated foot-and-mouth disease viral antigen(FMDV-Ag)were selected to prepare water adjuvant vaccines,ISA-206 oil adjuvant vaccine and adjuvant-free vaccine. ICR mice were immunized by muscle. Serum antibody level,lymphocyte proliferation response,and expressions of CD3+CD4+,CD3+CD8+ and CD4+CD44+,CD8+CD44+T cell subsets were measured to evaluate the immune enhancement effect after the vaccination,and the behaviors and weights of the mice were monitored for observing the safety of the vaccines. The results showed CDCP substantially enhanced the FMDV-specific antibody level(P<0.05)and significantly increased lymphocyte proliferative responses(P<0.05). T responses were also significantly enhanced(P<0.05),including proliferative responses of CD4+,CD8+,and CD44+ T cells. No abnormal behavior for mice after immunization was observed and the weights of mice also were not affected. CDCP as water adjuvant of FMDV-Ag has a fine immune-promoting effect by enhancing humoral and cellular immune response.

    Recent Advances in Plant Pep Peptide
    NIE Jia-yue, YANG Wen-wen, FAN Hong-xia, WANG You-ping, WU De-wei
    2021, 37(9):  219-225.  doi:10.13560/j.cnki.biotech.bull.1985.2021-0025
    Asbtract ( 614 )   HTML ( 26)   PDF (2465KB) ( 685 )  
    Figures and Tables | References | Related Articles | Metrics

    Plants harness many endogenous signal molecules,including hormones and polypeptides,to orchestrate developmental processes and responses to various environmental stresses. Peps are a class of 20-30 amino acid-long polypeptides that are widely distributed among angiosperms plant species. Peps can be perceived by plasma membrane-associated PEPR receptors and play important roles in multiple aspects of plant growth and defense. It has been reported that Peps are critical for plant defense against biological stresses such as pathogen infection and insect biting,as well as for plant tolerance to abiotic stresses such as high salinity. Moreover,Peps have also been shown to regulate the plant growth and development such as root elongation and leaf senescence. Here,we review recent advances in the generation of Peps,perception of receptor,signal transduction and other biological functions. We also discuss and prospect important unsolved scientific issues in Pep signaling and its potential applications in practice,aiming to provide reference for researchers in this field.

    Research Progress in Heat Shock Transcription Factors in Oryza sativa
    LIU Xiao-yi, YANG Jian, LIU Jing, WANG Bing, DAI Liang-ying, LI Wei
    2021, 37(9):  226-233.  doi:10.13560/j.cnki.biotech.bull.1985.2020-1460
    Asbtract ( 566 )   HTML ( 22)   PDF (1081KB) ( 778 )  
    Figures and Tables | References | Related Articles | Metrics

    Heat shock transcription factors(HSFs),as a type of transcription factor widely present in plants,are responsive to a variety of abiotic stresses. The HSFs at the end of the heat stress responses regulate the expressions of heat shock proteins by binding heat shock elements to forming a complex with other transcription factors,thereby participating in the regulation of plant resistance to abiotic stresses. Here,we summarized the typical structure of heat shock transcription factors in Oryza sativa,their functions and mechanisms in regulating resistance to abiotic and biotic stresses,and other functions in O. sativa,aiming to provide a reference for further dissecting the functions and mechanisms of HSFs and their application in breeding O. sativa’s resistance to abiotic and biotic stresses.

    Research Progress in miR159-GAMYB Regulating Plants Growth and Development
    LI Meng, CHEN Yue, HU Feng-rong
    2021, 37(9):  234-247.  doi:10.13560/j.cnki.biotech.bull.1985.2021-0028
    Asbtract ( 605 )   HTML ( 18)   PDF (1556KB) ( 691 )  
    Figures and Tables | References | Related Articles | Metrics

    miR159(microRNA159)is an ancient and conserved microRNA in plants,which targets a class of regulatory genes called GAMYB-like that encode R2R3 MYB transcription factors. miR159-GAMYB pathway is highly conserved,miR159 plays a critical role in vegetative growth,flowering induction,male reproduction,floral organ development,plant fertility,fruit development,seed sprouting and stress response by the post-transcriptional regulation of GAMYB. The article mainly discusses the influence of miR159-GAMYB pathway regulating plants growth and development,which provides a reference for future related researches.

    Recent Progress in Molecular Mechanism of Interaction Between Rice and Tilletia horrida
    JIANG Yu-qi, SHU Xin-yue, ZHENG Ai-ping, WANG Ai-jun
    2021, 37(9):  248-254.  doi:10.13560/j.cnki.biotech.bull.1985.2021-0075
    Asbtract ( 475 )   HTML ( 12)   PDF (2181KB) ( 432 )  
    Figures and Tables | References | Related Articles | Metrics

    Rice kernel smut(RKS)mainly damages rice floral organs in rice male sterile lines,and it occurs in most hybrid rice growing regions of the world,which becomes one of the main diseases to limit the seed production of hybrid rice. Studying the interaction mechanism between pathogenic bacteria and host may provide an important basis for mining gene resources of T. horrida and molecular breeding of rice with disease resistance. This review summarized the recent advances in infection processes,pathogenic related genes and biological pathways,and rice’s responses to T. horrida infection,as well as put forward future key research directions. The current studies demonstrated that the pathogen T. horrida invading the rice stigma at the earlier booting stage resulted in the disease. The lipid degradation and autophagy processes were key pathways of T. horrida successfully infecting the hosts. Genome sequencing and prediction of candidate effector proteins laid a foundation for the pathogenic genes researches of T. horrida. Furthermore,the report of sterile line resources resistant to T. horrida provided important resistant materials for gene mining and mechanism analysis. Future researches may focus on elucidating the pathogenic mechanism of T. horrida,mining the potential RKS-resistant genes using the resistance resources,and applying it in molecular breeding of resistance.

    Research Progress in the Effects of Genetically Modified Crops on Soil Microbial Community
    WANG Ting, YANG Yang, LI Jin-ping, DU Kun
    2021, 37(9):  255-265.  doi:10.13560/j.cnki.biotech.bull.1985.2020-1422
    Asbtract ( 499 )   HTML ( 22)   PDF (1129KB) ( 754 )  
    Figures and Tables | References | Related Articles | Metrics

    The safety of genetically modified crops is one of the hot issues in the world. Soil microorganisms play an essential role in soil organic matter transformation,nutrient mineralization,humus formation and soil structure improvement. Although transgenic crops have great potential in improving agricultural production,their potential effects on soil security and soil microorganisms need further in-depth study. This paper reviews the effects of insect-resistant,disease-resistant and herbicide-resistant genetically modified crops on soil microbial diversity and community structure. Then the paper puts forward the issues that should be considered in the evaluation of soil ecological security of transgenic crops,and makes a prospect for the future work.

    Application of CRISPR-Cas System in Nucleic Acid Detection
    HU Xiu-wen, LIU Hua, WANG Yu, TANG Xue-ming, WANG Jin-bin, ZENG Hai-juan, JIANG Wei, LI Hong
    2021, 37(9):  266-273.  doi:10.13560/j.cnki.biotech.bull.1985.2021-0015
    Asbtract ( 691 )   HTML ( 36)   PDF (1098KB) ( 1072 )  
    Figures and Tables | References | Related Articles | Metrics

    CRISPR-Cas is an efficient and practical gene editing tool,which is widely used in genome editing and regulation. Due to its high sensitivity and specificity,the CRISPR-Cas system also plays an important role in nucleic acid detection. It can detect pathogens,analyze single nucleotide polymorphisms(SNPs),and detect genetic mutations. The currently discovered Cas series nuclease tools open the door to the development of new strategies for different types of nucleic acid detection for various purposes. In addition,the CRISPR-Cas system is used in nucleic acid detection. The accuracy and efficiency of the field indirectly promote the progress of basic biology and applied biology research. The article outlines the latest developments and uses of different types of CRISPR-Cas systems that target specific nucleic acid detection,and the corresponding new generation of in vitro detection platforms,which provide new research ideas and theoretical basis for the field of nucleic acid detection.

    Construction and Application of a Drug Screening Method Based on PXR Promoter Reporter Gene
    SHEN Ya-li, PAN Yang-yang, WANG Jing-lei, MA Rui, ZHAO Gai-hong, WANG Gui-rong, ZHANG Qian, WANG Meng
    2021, 37(9):  274-284.  doi:10.13560/j.cnki.biotech.bull.1985.2020-1440
    Asbtract ( 492 )   HTML ( 17)   PDF (3983KB) ( 572 )  
    Figures and Tables | References | Related Articles | Metrics

    The objective of this work is to construct a drug screening method based on the dual luciferase reporter gene in mouse pregnane X receptor(mPXR)gene promoter,and to apply it for screening PXR inducers. Primers were designed by homologous recombination method. The mPXR gene promoter was amplified and recovered,and cloned into the pGL3-basic vector containing the luciferase reporter gene,thus the pGL3-basic-PXRpro recombinant plasmid was constructed. Then the recombinant plasmid and the internal reference plasmid pRL-TK were co-transfected into hepa 1-6 mouse hepatocarcinoma cells. Dexamethasone,a positive inducer of PXR in mice,was administered,and the inducing effect of PXR transcription activity was tested 24 h later. Further this method was used to screen PXR inducers from enrofloxacin,florfenicol and 10 natural products of Forsythia suspensa. The positive clones were obtained after monoclonal colony PCR,recombinant plasmids double enzymatic digestion and DNA sequencing. Finally the cloned fragment sequence was detected by the dual luciferase reporter system after transiently transfecting hepa 1-6 cells. The cloned fragment sequence had promoter activity,and the activity was induced by dexamethasone,and its relative activity was 0.112 9(P<0.05). Two antibacterial drugs and 9 natural products from F. suspense activated the PXR promoter using this reporter gene drug screening method. In conclusion,a drug screening method is successfully constructed for the mouse pGL3-basic-PXRpro reporter gene,which provides a tool for screening PXR inducers. Using this reporter gene method to screen natural products that induced PXR may provide candidate drugs for the prevention and treatment of diseases targeted by PXR.

    Others
    Content
    2021, 37(9):  285-285. 
    Asbtract ( 110 )   PDF (462KB) ( 75 )  
    Related Articles | Metrics
    Copyright
    2021, 37(9):  286-286. 
    Asbtract ( 92 )   PDF (145KB) ( 60 )  
    Related Articles | Metrics
    Cover
    2021, 37(9):  287-287. 
    Asbtract ( 83 )   PDF (1351KB) ( 60 )  
    Related Articles | Metrics