Ammonia emissions from soil under sheep grazing in Inner Mongolian grasslands of China

Ammonia (NH3) emission and redeposition play a major role in terrestrial nitrogen (N) cycles and can also cause environmental problems, such as changes in biodiversity, soil acidity, and eutrophication. Previous field grazing experiments showed inconsistent (positive, neutral, and negative) NH3 volatilization from soils in response to varying grazing intensities. However, it remains unclear whether, or to what extent, NH3 emissions from soil are affected by increasing grazing intensities in Inner Mongolian grasslands. Using a 5-year grazing experiment, we investigated the relationship between NH3 volatilization from soil and grazing pressure (0.0, 3.0, 6.0, and 9.0 sheep/hm2 ) from June to September of 2009 and 2010 via the vented-chamber method. The results show that soil NH3 volatilization was not significantly different at different grazing intensities in 2009, although it was higher at the highest stocking rate during 2010. There was no significant linear relationship between soil NH3 volatilization rates and soil NH4+-N, but soil NH3 volatilization rates were significantly related to soil water content and air temperature. Grazing intensities had no significant influence on soil NH3 volatilization. Soil NH3 emissions from June to September (grazing period), averaged over all grazing intensities, were 9.6±0.2 and 19.0±0.2 kg N/hm2 in 2009 and 2010, respectively. Moreover, linear equations describing monthly air temperature and precipitation showed a good fit to changes in soil NH3 emissions (r=0.506, P=0.014). Overall, grazing intensities had less influence than that of climatic factors on soil NH3 emissions. Our findings provide new insights into the effects of grazing on NH3 volatilization from soil in Inner Mongolian grasslands, and have important implications for understanding N cycles in grassland ecosystems and for estimating soil NH3 emissions on a regional scale.     

哈克尼西棉细胞质陆地棉雄性不育系orf160的克隆及遗传转化

以哈克尼西棉细胞质陆地棉雄性不育系及其保持系、恢复系为材料,克隆并测序线粒体功能基因的侧翼序列。通过生物信息学分析,在atp4下游发现一个哈克尼西棉细胞质不育系特有的orf160。orf160全长480 bp,N端与atp6序列部分同源,C端序列与核序列部分同源;编码的159个氨基酸序列,与膜蛋白、细胞周期蛋白具有部分同源性。以含有线粒体导肽的GFP瞬时表达载体转化后,在洋葱表皮的细胞膜上观察到绿色荧光,表明线粒体定向载体正常表达。构建酵母和植物表达载体,分别转化酵母、烟草和拟南芥;同对照相比,转基因酵母菌斑畸形且生长缓慢;转基因拟南芥和烟草种子大部分畸形,T₁和T₂代植株的结实率和种子发芽率降低,花粉着色比野生型浅。结果表明,orf160基因的表达影响受体正常发育。     

陆地棉叶绿体基因组密码子使用偏性的分析

 【目的】通过分析陆地棉叶绿体基因组密码子使用偏性,探讨影响密码子偏性形成的主要因素。【方法】以陆地棉叶绿体基因组为研究对象,利用Mobyle及其它软件,分析叶绿体50个基因密码子的使用模式。【结果】陆地棉叶绿体基因组密码子GC1与GC2之间的相关系数达到极显著水平,GC3含量最低,与GC12的相关系数为0.14,双尾检验未达到显著水平;单个基因ENC比值多分布在-0.05—0.05;对应性分析,第一轴上显示了10.27%的差异,与GC3、ENC的相关系数分别为0.223和-0.147,均未达到显著水平。【结论】陆地棉叶绿体密码子第三位偏好使用嘧啶,而最优密码子多以A或T结尾;密码子使用模式可能是由选择和突变及其它因素共同作用形成的。     

抗虫杂交棉棉铃性状与单铃重的关系分析

研究不同铃重类型抗虫杂交棉棉铃性状的差异及与铃重的相关性。以3个高品质陆地棉品种(系)和6个转基因抗虫棉品种(系)为亲本,配制36个正反交组合。以单铃重为聚类指标,采用最小离差平方和法和欧氏平方距离将36个杂交组合分为大、中、小铃3种类型。对3种类型的单铃重和11个棉铃性状进行了方差分析和相关分析。结果表明:单铃重、铃壳重、单铃皮棉重、单铃种子重、单铃种子数、单粒种子重、铃横径和铃纵径表现出显著或极显著差异,单粒种子纤维重、单铃不孕子数、不孕子率和铃壳率差异不显著;简单相关分析和偏相关分析表明多数棉铃性状与单铃重的相关性在不同铃重类型间表现基本相同。单铃种子重对单铃重的贡献最大,依次是单铃皮棉重、单铃种子数。铃壳率、单粒种子重、单粒种子纤维重、铃纵径与单铃重的简单相关和偏相关均未达到显著水平;大中铃重类型的铃重与铃壳重的简单相关系数达到显著水平。该研究结果对于通过棉铃性状的调控提高单铃重具有一定的指导意义。     

陆地棉胞质雄性不育系与保持系线粒体基因组RFLP分析

【目的】研究棉花细胞质雄性不育（CMS）系与保持系线粒体基因组的差异,为克隆棉花CMS基因奠定基础。【方法】以atpA, atp6, atp9, ccmB, ccmC, ccmFN1, cob, coxI, coxII, coxIII, matR, nad1bc, nad2, nad4, nad5, nad6, nad7c, nad9, rpl5, rrn18等20个线粒体基因保守序列为探针,利用Southern blot方法对棉花细胞质雄性不育系、保持系线粒体基因组进行RFLP分析。【结果】发现atpA、atp9、ccmB、nad1bc、nad6、nad7c、rrn18等基因在棉花不育系和保持系间存在明显RFLP多态性,其中atpA的差异最明显。【结论】atpA是棉花CMS系和保持系线粒体基因组间的一个明显差异位点。CMS系比保持系缺失一个rrn18拷贝。     

棉花DELLA蛋白GhGAI4b基因的克隆及功能初步分析

[目的]DELLA蛋白已在不同的物种中被广泛鉴定,并且功能区高度保守,是GA信号通路中的负调控因子,在棉花纤维细胞起始和发育中起重要的作用.研究棉花中DELLA基因的功能.[方法]从棉花中克隆DELLA蛋白的同源基因GhGAI4b,构建35S:GhGAI4b植物过量表达载体,通过农杆菌侵染花序的方法转入野生型拟南芥中,观察转基因植株的表型.[结果]序列分析表明GhGAI4b全长开放阅读框1 617 bp,编码538个氨基酸,具有DELLA蛋白的典型结构域.采用Clustal X将GhGAI4b的氨基酸序列与其他已知的DELLA蛋白氨基酸序列进行相似性比对,发现GhGAI4b氨基酸序列与陆地棉、拟南芥的相似性较高,达到47;以上.转基因株系表现出叶片颜色深绿,叶片缩小且肥厚,叶片细长呈锯齿状;有些株系会出现多重莲座叶,雄蕊发育不全,晚花,株高变矮等赤霉素不敏感突变体的表型.[结论]过表达GhGAI4b能够抑制拟南芥莲座叶的生长和发育,影响转基因拟南芥花器官的发育,严重时引起转基因拟南芥的不育.     

陆地棉单萜合酶基因GhTPS4和GhTPS5的克隆及棉铃虫诱导的基因表达分析

通过本地Blast筛选陆地棉转录组数据并结合RACE的方法,克隆获得2个陆地棉萜类合酶基因全长,命名为GhTPS4(Genebank登录号:KF860865)和GhTPS5(Genebank登录号:KF860866)。GhTPS4和GhTPS5基因分别编码590个和598个氨基酸,预测分子量为68.5 kDa和69.9 kDa,等电点为5.59和5.66。生物信息分析表明:GhTPS4和GhTPS5均含有叶绿体转运肽,并含有"DDxx D"和"RRX8W"保守域,具有典型的单萜合酶特征。此外,聚类分析表明GhTPS4和GhTPS5均属于Tpsb亚家族。利用荧光定量PCR的方法,对GhTPS4和GhTPS5在棉铃虫为害陆地棉(石远321)叶片的表达模式进行了研究,结果表明:GhTPS4的表达量在处理后不同时间均高于未处理对照,在处理后36 h表达量最高,表明GhTPS4在应答棉铃虫为害时可能起到了积极的防御作用。GhTPS5在处理后不同时间均比未处理对照表达量低,其功能尚需进一步探讨。     

Nonstructural carbohydrates in leaves subtending cotton bolls,fibers and embryos in response to nitrogen stress

A field study was conducted in 2013 and 2014 where cotton was exposed to three N regimes: (1) the control without N application (low N); (2) 260 kg N ha^?1 (medium N); (3) 520 kg N ha^?1 (high N). Boll size, lint mass per boll, seed mass per boll, fiber length and strength were significantly decreased under N deprivation in the two years. The increased carbohydrate levels of LSCB (leaf subtending the cotton boll) led to decreased carbohydrate levels of fibers in the low N relative to the other N treatments. The low N embryos exhibited lower starch concentrations at 17 and 31 DPA (days post anthesis), and TNC (total nonstructural carbohydrate) concentrations at 17, 31, 45 and 52 DPA compared to medium N embryos. Starch levels in LSCB had negative associations with those in fibers at 17, 31 and 45 DPA, but positive associations with those in embryos at 24 and 45 DPA. Fibers expressed negative associations with embryos in glucose level at 24 and 38 DPA, and in TNC levels at 17 and 45 DPA. The study suggests that carbon assimilate levels in fibers and embryos could explain the difference in boll yield components and fiber quality.     

Variation of pathotypes and races and their correlations with clonal lineages in Verticillium dahliae

Understanding pathogenic variation in plant pathogen populations is key for the development and use of host resistance for managing verticillium wilt diseases. A highly virulent defoliating (D) pathotype in Verticillium dahliae has previously been shown to occur only in one clonal lineage (lineage 1A). By contrast, no clear association has yet been shown for race 1 with clonal lineages. Race 1 carries the effector gene Ave1 and is avirulent on hosts that carry resistance gene Ve1 or its homologues. The hypothesis tested was that race 1 arose once in a single clonal lineage, which might be expected if V. dahliae acquired Ave1 by horizontal gene transfer from plants, as hypothesized previously. In a diverse sample of 195 V. dahliae isolates from nine clonal lineages, all race 1 isolates were present only in lineage 2A. Conversely, all lineage 2A isolates displayed the race 1 phenotype. Moreover, 900‐bp nucleotide sequences from Ave1 were identical among 27 lineage 2A isolates and identical to sequences from other V. dahliae race 1 isolates in GenBank. The finding of race 1 in a single clonal lineage, with identical Ave1 sequences, is consistent with the hypothesis that race 1 arose once in V. dahliae. Molecular markers and virulence assays also confirmed the well‐established finding that the D pathotype is found only in lineage 1A. Pathogenicity assays indicated that cotton and olive isolates of the D pathotype (lineage 1A) were highly virulent on cotton and olive, but had low virulence on tomato.