中植棉2号抗黄萎病的主基因+多基因遗传特性分析

以感病品种861为父本、抗病品种中植棉2号为母本配制杂交组合,构建6个世代群体(P_1、P_2、F_1、B_1、B_2和F_2),并在田间病圃进行抗病性鉴定,利用主基因￣多基因混合遗传模型的多世代联合分析法研究陆地棉抗黄萎病遗传特性。结果表明,中植棉2号抗性遗传符合E-1遗传模型,即2对加性￣显性￣上位性主基因+加性￣显性多基因遗传模型。2对主基因遗传以显性效应为主,且第2对主基因的显性效应比第1对主基因的显性效应大,多基因遗传以加性效应为主。B_1、B_2和F_2的主基因遗传率分别为68.24%、30.71%和82.09%,多基因遗传率分别为0、24.96%和0,环境方差占总表型方差的17.01%~44.33%。     

重病田和无病田棉花根组织中细菌群落结构的差异

为了研究黄萎病菌侵染条件下内生细菌群落结构多样性变化,以不同时期健康棉株和感染黄萎病菌(Verticillium dahliae)的棉株根部组织为材料,提取样品总DNA,扩增内生细菌16S r DNA的V3可变区,将扩增产物进行变性梯度凝胶电泳分析,并对21个优势条带进行切胶回收、克隆测序,构建系统发育树。结果表明:不同时期发病棉株和健康棉株样品可以聚成2大类,一是苗期和蕾期的根部样品,二是初花期和结铃期的根部样品;感染黄萎病棉株内生细菌种类多样性下降;BLAST比对结果发现21个克隆序列与Gen Bank数据库中已知16S r DNA基因序列同源性很高(99%~100%),绝大多数属于变形菌门(Proteobacteria),占总克隆序列的80.95%,其他不可培养细菌占19.05%。说明黄萎病菌的侵染能明显影响内生细菌群落结构多样性。     

Targeted transfer of trait for Verticillium wilt resistance from Gossypium barbadense into G. hirsutum using SSR markers

Verticillium wilt (VW) is a soil‐borne disease of cotton that is destructive worldwide. Transferring desired traits from Gossypium barbadense is challenging through traditional interspecific introgression. We previously demonstrated that a molecular marker, BNL3255‐208, is associated with VW resistance in G. barbadense. This breakthrough opens the way for marker‐assisted selection (MAS) breeding. Here, the highly resistant G. barbadense cv. ‘Pima90‐53’ and the severe diseased Gossypium hirsutum cv. ‘CCRI8’ were used as donor parent and recipient parent, respectively. Our goal was to transfer the disease resistance from donor to recipient via MAS. Among 71 MAS obtained lines, as many as 19 lines had enhanced resistance. Among those lines, 11 lines showed high resistance and four lines displayed resistance to VW. Moreover, seven lines displayed improved fibre quality. After combining the markedly improved resistance and fibre properties, we identified two elite innovated introgression lines – ZY2 and ZY31 – that did not seem to differ in other agronomic traits from the recipient parent. This study first successfully transferred of G. barbadense resistance into G. hirsutum by MAS.     

GFP标记的马铃薯大丽轮枝菌生物学特性研究

为了研究马铃薯黄萎病菌的侵染机制,将绿色荧光蛋白基因(GFP)通过农杆菌介导的遗传转化方法转入马铃薯黄萎病菌VD012中,经过潮霉素选择性培养基的筛选和分子鉴定,获得了47株有绿色荧光信号的阳性转化子。随机挑取8株阳性转化子,以野生型菌株为对照,对转化子的菌落形态、菌丝的生长速率、产孢量、粗毒素含量和致病力进行了研究。结果表明,8株阳性转化子中有3株微菌核产生的数量明显高于野生型,1株转化子微菌核产生量低于野生型菌株;各阳性转化子的生长速率和野生型菌株差异不显著;阳性转化子的产孢量与对照相比,有2株差异不显著。其余均有不同程度的降低。保湿培养8 h,所有阳性转化子的平均萌发率低于野生型菌株。相比对照,粗毒素的含量表现为升高趋势的转化子有7株,1株表现为下降趋势。致病力测定的结果表明,致病力增强的转化子有1株,2株转化子的致病力呈现降低的趋势。     

我国棉花黄萎病菌致病力分化及ISSR指纹分析

 来自我国12个省84个县（市）的棉花黄萎病菌,在PDA培养基上存在5种不同的培养类型,其中,产生微菌核较多的B型菌株为优势类群,占72.9%。长江流域的菌株培养性状变异最大,新疆棉区的变异最小。致病力测定结果和ISSR指纹图谱均将167个单孢菌株划分为强、弱、中3个致病力类型,供试菌株的ISSR指纹图谱与菌株的致病力存在明显的相关性。中等致病力类型菌株在我国占主导地位;强致病力类型的菌株主要分布在河北、河南、湖北等省;弱致病力类型菌株主要分布在新疆和江苏。     

大丽轮枝菌（Verticillium dahliae VdLs.17）分泌组预测及分析

 【目的】预测并分析大丽轮枝菌基因组范围内的分泌蛋白,为大丽轮枝菌分泌蛋白致病机理的研究奠定基础。【方法】利用已公布的大丽轮枝菌全基因组序列,组合使用生物信息学软件SignalP、TargetP、TMHMM、Big-pi和PROSITE,预测大丽轮枝菌基因组范围内所有分泌蛋白,定义为分泌组。统计分析分泌组中蛋白N-端信号肽特点;应用碳水化合物活性酶类数据库和病原菌-寄主互作蛋白数据库对分泌组蛋白进行注释,预测分泌组中潜在果胶酶、纤维素酶和病原菌寄主互作蛋白;利用真菌激发子的保守结构域,预测分泌组中潜在的激发子蛋白集;应用BLASTP程序比较分析大丽轮枝菌和黑白轮枝菌分泌组,获得大丽轮枝菌相对于黑白轮枝菌特异的分泌蛋白。【结果】大丽轮枝菌分泌组共有922个蛋白。信号肽分析表明,以19个氨基酸为信号肽的蛋白数目最多,非极性氨基酸丙氨酸的出现频率最高,而有带电侧链的氨基酸天冬氨酸和谷氨酸的出现频率最低,信号肽的-3和-1位置上的氨基酸相对保守。大丽轮枝菌分泌组含有158个潜在的碳水化合物活性酶类,其中,包括10个果胶水解酶和14个果胶裂解酶;190个潜在的病原菌-寄主互作蛋白、97个含有RxLx[EDQ]模体的蛋白和52个富含半胱氨酸的小分子量分泌蛋白;58个相对于黑白轮枝菌分泌组特异的蛋白。【结论】本文建立了预测大丽轮枝菌分泌组蛋白的方法。分泌组蛋白信号肽长度具有高度的变异性,氨基酸组成多为脂肪族氨基酸,序列在C-端结构域较为保守。分泌组中包含大量潜在的果胶降解酶、病原菌-寄主互作蛋白、RxLx[EDQ]模体蛋白和富含半胱氨酸的小分子量蛋白等致病相关蛋白。     

大丽轮枝菌拮抗细菌菌株12-51的筛选鉴定与抗菌物性质分析

由大丽轮枝菌（Verticillium dahliae Kleb）引起的棉花黄萎病,是一种危害严重的世界性病害。通过初筛和复筛得到了对棉花黄萎病具有较高拮抗活性的细菌菌株12-51,通过对其进行形态鉴定、生理生化特征鉴定和16S rDNA全序列分析,最终鉴定此菌株为Bacillus velezensis,16S rDNA序列相似度达99.74%。采用有机溶剂萃取和盐析法对发酵产物进行了提取,初步断定拮抗物质为蛋白类。通过对粗蛋白进行进一步研究发现,60%组分活性较高,并且对热以及酸碱敏感性较小。试验结果为棉花黄萎病的生物防治奠定了基础。     

蜡蚧轮枝菌对平菇厉眼蕈蚊幼虫和蛹的毒力测定

摘 要: 采用蜡蚧轮枝菌(Verticillium lecanii) MZ041024菌株不同分生孢子浓度对平菇厉眼蕈蚊的幼虫和蛹进行毒力测定。结果表明,MZ041024菌株杀虫活性较高,对幼虫和蛹最高校正死亡率分别为91.79%和88.94%,LC50分别达到1.745×105个/ml和4.723×105个/ml;LT50最短分别为3.51d和3.78d 。     

The potential for early generation selection to identify potato clones with resistance to Verticillium wilt

Verticillium wilt (VW) of potato, caused primarily by the fungus Verticillium dahliae, results in yield loss and is therefore an important soil-borne disease. Resistance to VW exists in potato germplasm and is used by breeders during cultivar development. Breeders could make more rapid progress toward the development of VW resistant clones if they had an effective early generation selection strategy. The purpose of this study was to determine whether selection for VW resistance could be carried out in the first tuber generation on single hills. One hundred and fifty-two clones from 19 families were planted as single hills on a V. dahliae-infested field. Each plant was scored for vine maturity, VW symptom expression, yield, stem colonization (colony forming units (cfu), in dried basal stem segments) and incidence (percent infected stems). In the second clonal generation, which consisted of replicated four-hill plots, stem colonization scores and incidence values were used to identify clones which were more resistant than a moderately resistant cultivar and others which were more susceptible than a susceptible cultivar. The efficiency and reliability of the single-hill selection strategy, based on symptoms and yield, was then determined by comparison to the four-hill results. We determined that the best single-hill selection strategy was negative selection (discard clones with the lowest performance) with low stringency, based on yield.     

棉花黄萎病拮抗细菌12-47发酵条件的优化

本实验采用单因素实验的方法在摇床培养的基础上对影响棉花黄萎病拮抗细菌12-47菌株产生拮抗物质的营养因素进行了考察,其中包括发酵培养基的C源、N源、无机盐等。在此基础上通过正交试验对各因素的浓度和组合进行了优化;通过L16（43）正交实验对培养最佳培养基起始pH值、摇瓶装液量、接种量、发酵时间等条件进行了摸索。最终确定优化后的培养基为：玉米粉3%、大豆蛋白胨5%、Mn2+ 0.1%、K+ 0.05%、起始pH8.0、装液量100mL/250mL三角瓶、接种量8%、发酵时间48h。用优化后的培养基进行发酵,发酵液抑菌活性比优化前提高了269%。