干旱胁迫下喷施外源植物激素对甘薯生理特性和产量的影响

为探讨不同时期干旱胁迫下喷施外源植物激素对甘薯生理特性和产量的影响,明确喷施外源植物激素的最佳时期,在人工控水条件下研究了移栽后20(前期)、60(中期)和100 d(后期)干旱胁迫下喷施6-苄氨基嘌呤(6-BA)、α-萘乙酸(NAA)和脱落酸(ABA)对甘薯内源激素含量、光合荧光特性和产量的影响。结果表明: 与喷清水相比,喷施外源植物激素均能显著提高甘薯产量,以6-BA增幅最大,其次是NAA和ABA;前期喷施效果好于中期和后期。不同时期干旱胁迫下,喷施外源植物激素可显著提高甘薯叶片的光合和叶绿素荧光参数,缓解因干旱引起的玉米素核糖核苷(ZR)和生长素(IAA)含量下降的现象。逐步回归分析表明,内源激素和光合特性是影响甘薯产量的关键指标。通径分析表明,前期干旱胁迫下喷施外源植物激素主要通过影响净光合速率(P_n)、IAA、ZR、最大光化学效率和光化学性能指数来影响甘薯产量。生长前期喷施6-BA可调控甘薯内源激素含量,提高光合特性,有效缓解干旱造成的产量损失。     

外源砷胁迫对两种土壤中细菌和古菌群落的影响

采矿和冶炼等活动会导致土壤中砷的累积,给农产品质量安全和土壤微生物带来不利影响。本文研究了外源砷进入黄壤(YS)和紫色砂页岩发育土壤(RS)后有效砷含量随时间的变化,并采用MiSeq高通量测序研究土壤中细菌和古菌在未加砷和外源砷胁迫1、30、360 d后的群落变化,探讨砷胁迫下土壤细菌和古菌的适应机制。结果表明: 外源砷进入土壤后,土壤有效砷含量随时间推移逐渐降低,并显著影响土壤细菌和古菌的群落组成。土壤细菌的优势菌群丰度变化显著;而古菌中仅丰度较低的菌群发生显著改变,优势菌群丰度变化较小,推测古菌群落具有高耐砷性和稳定性。与砷胁迫时间相比,土壤砷的有效性对细菌和古菌群落结构的影响更大。研究结果可为砷污染农田的安全利用及微生物修复等提供参考。     

Overexpression of the chitinase gene CmCH1 from Coniothyrium minitans renders enhanced resistance to Sclerotinia sclerotiorum in soybean

Transgenic Research - Pathogenic fungi represent one of the major biotic stresses for soybean production across the world. Sclerotinia sclerotiorum, the causal agent of Sclerotinia stem rot, is a...     

圈养林麝麝香分泌时间节律及影响因素

在2016年和2017年的5—8月,我们对川西马尔康麝场圈养林麝（Moschus berezovskii）的麝香分泌进行了行为与生理监测,对麝香分泌的各阶段进行了准确判定,记录了泌香启动、泌香盛期开始、泌香盛期终止及泌香结束的时间阶段及持续时间长,分析了林麝麝香分泌的时间阶段与体重和年龄等因素的关系。结果表明,马尔康麝场的雄性林麝平均泌香启动日在6月16日[（167.06±7.75）d,n=141],于6月17日进入泌香盛期[（168.52±7.67）d,n=141],6月21日[（172.17±7.26）d, n=138]泌香活动减弱,至6月25日[（176.27±8.11）d, n=131]泌香结束;雄麝体重与其泌香启动、泌香盛期停止及泌香结束时间呈显著负相关（r_PS = -0.234,P_PS =0.028;r _VSF = -0.215,P_VSF = 0.047;r _SE = -0.229,P_SE = 0.043）,即雄麝的体重越大,其泌香越早;各年龄组间的平均泌香时长差异显著（F _{17, 113} = 3.482, P = 0.003）,其中2岁雄麝平均泌香时长最长[（13.07±2.08）d, n=20],显著高于3岁[（9.38±0.76）d, n=12, P = 0.042]和4岁[（7.80±1.60）d, n=5, P = 0.013]个体;马尔康林麝平均泌香量为（11.85g±0.96）g, （n=114）,随泌香时长延长有增加趋势,但不显著（P = 0.854）。基于上述林麝雄体的泌香时间、泌香量与年龄和体重等因素间的关联,可对圈养的林麝个体间的泌香力、泌香量等进行区分和预判,作为圈养林麝驯养生产力优化的依据,并可为圈养林麝优质品系的选育提供参考。     

地涌金莲MlCCD8b基因的克隆与表达分析

该研究以地涌金莲(黄色苞片型YN01和红色苞片型RD05)为材料,采用RACE技术克隆获得地涌金莲CCD8b基因的cDNA全长,进行氨基酸序列比对及系统进化树构建,并采用实时荧光定量PCR技术检测MlCCD8b基因在不同地涌金莲类型和不同组织中的表达模式。结果表明：(1)序列分析显示,MlCCD8b的ORF全长1 671 bp,编码556个氨基酸,存在1个类胡萝卜素加氧酶家族的典型保守结构域RPE65,推测其相对分子量为61 574.26 Da,等电点6.61,亚细胞定位于叶绿体基质中的类囊体上,所编码的MlCCD8b蛋白为亲水性蛋白。(2)同源对比分析及构建系统进化树发现,地涌金莲MlCCD8b蛋白与小果野蕉亚种、凤梨等单子叶植物的CCD8b蛋白遗传关系最近。(3)荧光定量PCR检测结果表明,MlCCD8b在吸芽数量多的黄色苞片型YN01的所有组织中均有表达,而在吸芽数量少的红色苞片型RD05中,其苞片内未能检测到MlCCD8b的表达,但其他组织中皆有表达;MlCCD8b在2种类型地涌金莲中呈现一致的组织表达特异性,即在花序轴中的表达量最高,其次是吸芽芽点、根尖和叶片,在苞片中的表达量最低或不表达。(4)2种类型地涌金莲同一组织部位比较结果显示,RD05的花序轴、吸芽芽点、根尖和叶片中的MlCCD8b相对表达量分别是YN01的4.47、4.67、2.09和1.10倍。(5)利用高效液相色谱 串联质谱法测得RD05根尖部位的5 脱氧独脚金醇含量是YN01的15.57倍,与MlCCD8b在根尖的表达趋势一致。研究认为,MlCCD8b基因可能通过调控独角金内酯的合成,促进或抑制地涌金莲吸芽的萌生。该研究结果可为MlCCD8b基因的生物学功能研究提供依据,并为今后通过分子辅助育种调控MlCCD8b的表达,从而控制地涌金莲吸芽数量提供理论支持。     

有机物料还田对双季稻田土壤有机碳及其活性组分的影响

有机物料还田是提升农田土壤有机碳、培肥土壤的重要措施。为探讨不同有机物料的还田效果,采用室外培养方法,研究了在等碳输入条件下,施用水稻秸秆、紫云英、生物有机肥、猪粪和水稻秸秆生物炭对洞庭湖双季稻区潮土有机碳和活性有机碳组分含量的影响。结果表明: 经过180 d的培养试验,与不施用有机物料相比,施用有机物料提高了土壤活性有机碳含量。生物有机肥、猪粪和水稻秸秆生物炭处理分别使土壤有机碳含量显著提升了26.1%、9.7%和30.7%,水稻秸秆和紫云英对土壤有机碳含量的提升效应在试验期间并不显著。水稻秸秆和紫云英还田更有利于土壤可溶性有机碳和微生物生物量碳的积累,猪粪更有利于土壤可溶性有机碳的积累,生物有机肥更有利于土壤微生物生物量碳和易氧化有机碳的积累,水稻秸秆生物炭则更有利于土壤微生物生物量碳和轻组有机碳的积累。与水稻秸秆还田相比,紫云英、生物有机肥、猪粪和水稻秸秆生物炭还田使土壤碳库管理指数分别提高了31.8%、111.6%、62.2%和50.7%。从土壤固碳和土壤碳库管理指数来看,生物有机肥、猪粪和水稻秸秆生物炭的还田效果优于水稻秸秆和紫云英还田。     

Rapamycin inhibits AR signaling pathway in prostate cancer by interacting with the FK1 domain of FKBP51

Reactivation of the androgen receptor signaling pathway in the emasculated environment is the main reason for the occurrence of castration-resistant prostate cancer (CRPC). The immunophilin FKBP51, as a co-chaperone protein, together with Hsp90 help the correct folding of AR. Rapamycin is a known small-molecule inhibitor of FKBP51, but its effect on the FKBP51/AR signaling pathway is not clear. In this study, the interaction mechanism between FKBP51 and rapamycin was investigated using steady-state fluorescence quenching, X-ray crystallization, MTT assay, and qRT-PCR. Steady-state fluorescence quenching assay showed that rapamycin could interact with FKBP51. The crystal of the rapamycin-FKBP51 complex indicated that rapamycin occupies the hydrophobic binding pocket of FK1 domain which is vital for AR activity. The residues involving rapamycin binding are mainly hydrophobic and may overlap with the AR interaction site. Further assays showed that rapamycin could inhibit the androgen-dependent growth of human prostate cancer cells by down-regulating the expression levels of AR activated downstream genes. Taken together, our study demonstrates that rapamycin suppresses AR signaling pathway by interfering with the interaction between AR and FKBP51. The results of this study not only can provide useful information about the interaction mechanism between rapamycin and FKBP51, but also can provide new clues for the treatment of prostate cancer and castration-resistant prostate cancer.     

Integration of high-throughput analytics and cell imaging enables direct early productivity and product quality assessment during Chinese Hamster ovary cell line development for a complex multi-subunit vaccine antigen

Mammalian cell line generation typically includes stable pool generation, single cell cloning and several rounds of clone selection based on cell growth, productivity and product quality criteria. Individual clone expansion and phenotype-based ranking is performed initially for hundreds or thousands of mini-scale cultures, representing the major operational challenge during cell line development. Automated cell culture and analytics systems have been developed to enable high complexity clone selection workflows; while ensuring traceability, safety, and quality of cell lines intended for biopharmaceutical applications. Here we show that comprehensive and quantitative assessment of cell growth, productivity, and product quality attributes are feasible at the 200–1,200 cell colony stage, within 14 days of the single cell cloning in static 96-well plate culture. The early cell line characterization performed prior to the clone expansion in suspension culture can be used for a single-step, direct selection of high quality clones. Such clones were comparable, both in terms of productivity and critical quality attributes (CQAs), to the top-ranked clones identified using an established iterative clone screening approach. Using a complex, multi-subunit antigen as a model protein, we observed stable CQA profiles independently of the cell culture format during the clonal expansion as well as in the batch and fed-batch processes. In conclusion, we propose an accelerated clone selection approach that can be readily incorporated into various cell line development workstreams, leading to significant reduction of the project timelines and resource requirements.     

Revealing mechanism of Caulis Sargentodoxae for the treatment of ulcerative colitis based on network pharmacology approach

Objective: The traditional Chinese medicine Caulis Sargentodoxae is widely used in the treatment of ulcerative colitis (UC), but the mechanism remains unknown. The present study aims to reveal its effective components, targets and pathways through network pharmacology and bioinformatics approaches.Materials and methods: Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) was used to identify effective components. The ligand-based targets prediction was achieved through SwissTargetPrediction and TargetNet. UC-related targets were identified using Gene Expression Omnibus (GEO) data and DisGeNET. The common targets of disease and components were constructed and analyzed by PPI network. Lastly, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses are used to explain the functions of these common targets. Components-Targets-Pathways network was visualized and analyzed to further reveal the connection between the components and targets.Results: Eight active components and 102 key targets were identified to play an important role in UC. These targets were related to regulation of protein serine/threonine kinase activity, positive regulation of cell motility, response to molecule of bacterial origin, response to toxic substance, ERK1 and ERK2 cascade, peptidyl-tyrosine modification, inositol lipid-mediated signaling, cellular response to drug, regulation of inflammatory response and leukocyte migration. Moreover, HIF-1 signaling pathway and PI3K-Akt signaling pathway were the key targets involved in UC-related signaling pathways.Conclusion: The eight active components of Caulis Sargentodoxae mainly play a therapeutic role for UC through synergistic regulation of HIF-1 signaling pathway and PI3K-Akt signaling pathway.