棉花抗黄萎病QTL初步定位

本研究以高抗黄萎病的海岛棉品种海7124和高感黄萎病的陆地棉品种邯郸14组配的F2群体184个株系,构建了一个分子标记遗传连锁图,包括了142个位点和30个连锁群,标记间的平均距离为8.24cM,全长1169.6cM,覆盖棉花总基因组约23.4%。用复合区间作图分析共检测到12个抗黄萎病相关QTL,其中田间抗病性检测到不同时期7个病情指数相关QTL,1个病株率相关QTL,温室苗期抗病性定位4个病株率相关QTL。从绝对量上看,各QTL加性效应从2.20到42.39,显性效应从1.65到32.25,解释表型变异1.09%～22.73%,且田间抗病性获得的QTL与温室苗期抗病性得到的QTL基本相同,其中qFDI711-30-0.01位点距MUSS294仅为0.01cM,加性效应较高解释表型变异22.32%,这对于培育优质抗病材料用于标记辅助育种具有一定的参考价值。     

棉花品种对黄萎病慢病性初步研究

2002-2003年在河南省新乡县李台基地重病田,通过定株定时调查发病情况和各单株的产量结构,研究了不同抗病性棉花品种对黄萎病慢萎性。结果表明,若单纯以8月下旬的病情指数为标准,不少品种的抗病性均不理想,但从黄萎病发展历程分析,则发现有一部分耐至感病品种,其病情发展缓慢,对产量的影响也达不到显著水平。为此,提出棉花品种存在对黄萎病的慢病性,简称棉花品种慢萎性。     

转基因抗虫棉抗黄萎病鉴定及黄萎病发生规律

采用田间病圃鉴定法,研究了国内外45个转基因抗虫棉品种(系)的黄萎病抗病性,结果表明,以相对抗性指数(IR)划分抗性类型,抗病品种(系)占6.7％,耐病品种(系)占51.1％,感病品种(系)占42.2％。美国引进品种(系)比国内自育品种(系)的抗病性稍差。棉花黄萎病菌在不同转基因抗虫棉品种(系)上的侵染、扩展规律有较大差异,可分为5类。不同品种(系)的抗病特性不同,可分为3大类。此外,筛选出抗虫棉抗病性鉴定的对照品系—J103。     

棉花远缘杂交创制抗黄萎病新种质

本研究通过陆地棉与异常棉、辣根棉、旱地棉、雷蒙德氏棉、黄褐棉等野生棉的远缘杂交和回交等,育成65份陆地棉背景的远缘种质系。农艺性状鉴定结果 ,这些种质系的株高平均值与陆地棉品种处于相同水平,单株果枝数、单株果节数、单株结铃数、铃重、衣分平均值均低于陆地棉品种,纤维长度、断裂比强度均高于陆地棉品种,马克隆值均低于陆地棉品种。落叶型黄萎病菌系V991人工病圃抗性鉴定结果,从65份陆地棉背景的远缘种质系中获得5份抗黄萎病的棉花新种质系,其中苏远040、苏远045、苏远061病情指数分别为8.33、4.35、7.79,高抗黄萎病;苏远030、苏远034病情指数分别为12.35、13.70,抗黄萎病。追踪陆地棉抗黄萎病新种质系的亲缘关系发现,苏远040和苏远045为陆地棉与雷蒙德氏棉杂交后代,苏远061为陆地棉与黄褐棉杂交后代,苏远030和苏远034为陆地棉与旱地棉杂交后代。这些优质、抗黄萎病新种质的育成,拓宽了陆地棉的遗传基础,为我国棉花抗病新品种选育提供种质材料。     

中棉所12的黄萎病抗性遗传与育种应用研究

 以2个海岛棉品种和5个陆地棉品种为材料与中棉所12进行正反交,配制14个杂交组合的F₁和F₂ 。采用纸钵育苗,撕底伤根接种方法对14个组合的F₁和F₂群体进行黄萎病抗性鉴定。结果表明,以中棉所12作父本与海岛棉抗黄萎病品种或陆地棉抗黄萎病品种进行杂交,F₂抗（耐）病株与感病株的分离符合3：1的分离规律,说明海岛棉的抗黄萎病性对于中棉所12的耐黄萎病性为显性,中棉所12的耐黄萎病性对于陆地棉的感黄萎病性为显性,控制黄萎病抗性的基因为一个显性主基因。然而,以中棉所12为母本与海岛棉品种、抗病陆地棉品种和感病陆地棉品种进行杂交,F₂群体中90%以上的个体为抗病类型,说明中棉所12的细胞质中存在着抗黄萎病的遗传成分,具有细胞质母体遗传的特点,在棉花抗黄萎病育种中具有重要的利用价值。     

棉花抗黄萎病鉴定与评价标准的商榷

【目的】通过系统研究黄萎病抗病鉴定的关键技术和评价标准,解决现有棉花品种黄萎病抗性评价标准中存在的问题,建立新的田间棉花黄萎病分级标准和评价标准。【方法】以14个2014年国家主推棉花品种为试验材料,从接种量、可靠样本量、病情分级和评价方法 4个方面系统研究黄萎病抗性鉴定的关键技术和评价标准。【结果】确定了小区接种和行沟接种的最佳接菌量分别为60 g·m-2和10 g·m-1,小区调查最低样本量为50株;制定了新的分级标准为0级(0%)、1级(1%~33%)、2级(34%~66%)、3级(67%~99%)和4级(100%)。连续2年利用相对病程发病面积和发病高峰期的相对病情指数2种评价方法判定品种抗病性,前者作为抗病性评价的依据更为科学可靠,一致率达到100%。【结论】本试验为完善抗病鉴定技术操作规程,规范抗性评价方法,促进棉花抗病品种的选育、审定和推广提供了科学依据。     

棉花黄萎病菌与品种互作的格局分析

 以23个棉花品种(系)接种18个黄萎病菌株的抗感表现(病情指数)为分析对象,在聚类分析的基础上用双标图方法研究了寄主 病原的互作格局。结果表明：23个棉花品种分为6个抗病性不同的品种组,18个菌株聚为5个致病力不同的菌株群。5031和50312组成的品种组G1对所有菌株群具有最高抗性,可作为抗黄育种的高抗质源。来自河北成安的VD25与来自江苏常熟的XS4、XS6和XS7组成的菌株群S4与G5品种组存在互作,其致病力最强,是棉花黄萎病的优势菌株群,在抗黄育种和抗黄鉴定中应予以高度重视。S2和S3菌株群与56%的参试品种构成的3个品种组间存在互作,是潜在的优势菌株群。具平均抗病性的G2品种组是黄萎病菌的优良寄主。     

新疆棉花黄萎病新型菌株NH1致病性鉴定及抗病分析

为抗病育种提供新的强致病力资源菌株,分析新疆主要产棉区棉花黄萎病病原菌的形态特征、菌系变异和致病力分化情况,探究菌株的抗病水平。本研究从新疆南、北疆植棉区共采集35株棉花黄萎病病杆,经分离纯化、形态鉴定及菌落观察,筛选出1株致病力较强的菌株NH1作为供试菌株。采用鉴别寄主法,检测发现NH1菌株对20个不同抗性的寄主品种均有表现不同程度的致病力。抗性鉴定结果显示,参与鉴别寄主的20个品种材料中未出现免疫或高抗的材料,表现抗病的材料有5个,占鉴定材料的25%;感病品种11个,占鉴定材料的55%;耐病的材料4个,占鉴定材料的20%。根据病情指数进行划分,NH1菌株的相对病情指数为37.1,大于30.1,生理型鉴定为1号生理小种,PCR鉴定为落叶型。结果表明,NH1菌株为生理型1型的强致病力落叶型菌株,可作为新的强致病力菌,对提升棉花黄萎病的统防统治具有重要实践意义。     

我国棉花品种抗黄萎病鉴定存在的问题及对策

对１９９０～１９９６年通过省级审定的８４个棉花品种的黄萎病病指分析表明,大多数品种的病指与其实际抗病性不符。分析其原因认为,抗性鉴定的病圃及鉴定方法不规范,品种审定时对抗黄萎病性能无明确要求等所造成,为此提出解决问题的对策。     

棉花黄萎病菌致病力分化与寄主抗病性遗传研究进展

棉花黄萎病是世界各主要产棉国家和地区普遍发生和危害最重的病害之一，本文综述了棉花黄萎病菌致病力分化与寄主抗病性遗传研究进展。病菌种内存在着明显的致病力分化。寄主抗病性的遗传在种间和种内材料间亦存有差异。为了定向而有效地培育抗病品种、合理进行品种布局，对此类问题仍需进一步研究和探讨。     

转几丁质酶和葡聚糖酶双价基因棉花株系对黄萎病的抗性

 采用人工病圃、重病田及苗期室内鉴定相结合的方法,研究了转双价（Chi+Glu）基因的棉花株系61217-1对棉花黄萎病的抗性。结果表明,转基因棉花株系61217-1在人工病圃连续3年达抗病水平,病指极显著低于其受体中棉所24。2009年该转基因株系在河南安阳、江苏大丰及新疆石河子棉花黄萎病重病田的病指分别为18.7、18.1和16.7,均极显著低于受体中棉所24的病指。该转基因株系对来自河北、湖北、新疆的3个强致病力落叶型菌株均达抗病水平,病指分别为16.4、16.8和15.6,也极显著低于受体中棉所24的病指。表明转双价（Chi+Glu）基因的棉花株系61217-1具有优异稳定的抗黄萎病性。     

Physiological Mechanism of Different Varieties and Potassium Application Amounts on Cotton Resistance to Verticillium Wilt

[Objective] It has theoretical guiding significance for the comprehensive control technology of Verticillium wilt of cotton to explore the effect of potassium nutrition on mechanism of cotton Verticillium wilt resistance under the condition of pathogen stress of Verticillium dahliae. [Method] Three cotton cultivars Jimian 11 (JM-11 as susceptible cultivar), Zhongzhimian 2 (ZZM-2 as control cultivar), Nongda 601 (ND-601 as resistant cultivar) were adopted, and each cultivar had 0, 75, 150, 225, 300 kg· hm－2 potassium levels represented by K0, K75, K150, K225, K300. Two factors split plot experiment was designed under artificial nursery inoculated Verticillium dahlia, to observe disease index of Verticillium wilt, and researched resistance associated substances content and enzyme activity. [Result] (1) The lignin, total phenol contents and POD, PAL activity in leaves or roots of JM-11 were more greatly influenced by Verticillium wilt compared with ZZM-2, ND-601; the crucial disease resistance substances and enzymes were lignin, total phenol and PAL for JM-11, total phenol for ZZM-2 and ND-601. (2) The lignin, total phenol contents and POD, PAL activities of JM-11 were more sensitive to root potassium than ND-601, but weaker than that of ZZM-2. The sensitivity of POD, PAL activities in functional leaves of JM-11 to potassium were more sensitive than ND-601 and ZZM-2. (3) The best effect of potash fertilizer for relieving disease index for JM-11, ZZM-2, ND-601 were 32.5%, 26.4%, 16.5%, and corresponding potassium application amounts were 290.5 kg·hm－2, 253.5 kg·hm－2 and 229.5 kg·hm－2, respectively. [Conclusion] Application of potassium could increase the resistance to Verticillium wilt and reduce the occurrence of cotton Verticillium wilt.     

Creation of a New Resistant Germplasm to Verticillium Wilt by Distant Hybridization in Upland Cotton

To create new germplasm lines resistant to Verticillium wilt in upland cotton, 65 distant hybridization germplasm lines (DHGLs) in upland cotton genetic background were cultivated by interspecific hybridization between Gossypium hirsutum and wild species including G. anomalum, G. armourianum, G. aridum, G. raimondii, G. mustelinum, interspecific F₁ backcrossing with G. hirsutum for four generations, and selfing for four generations, followed by a conventional breeding program. The results of agronomic trait identification during 2011-2012 indicated that average plant heights of DHGLs were closely similar to commercial cultivars of upland cotton, while average fruit branches, fruit nodes, bolls of DHGLs individual plant were lower than those in commercial cultivars of upland cotton. Average single boll weight and lint percentage of DHGLs were lower than commercial cultivars of upland cotton. Fiber length, strength, fineness and maturity of DHGLs were reasonably collocated. Fiber of most lines was suitable for spinning extra high count yarn, but the main fiber quality indices of commercial cultivars of upland cotton were not well coordinated. Identification of resistance to Verticillium wilt in defoliation disease nursery during 2012-2013 indicated that five DHGLs resistant to Verticillium wilt . Suyuan 040, Suyuan 045 and Suyuan 061 were highly resistant to Verticillium wilt with a disease index of 8.33, 4.35 and 7.79, respectively. Suyuan 030 and Suyuan 034 were resistant to Verticillium wilt with a disease index of 12.35 and 13.70, respectively. The genetic relationship of new germplasm lines resistant to Verticillium wilt were traced and showed that Suyuan 040 and Suyuan 045 were DHGLs of G. raimondii, Suyuan 061 was DHGL of G.mustelinum, Suyuan 030 and Suyuan 034 were DHGLs of G. aridum.     

转CpTI基因对棉花主要病害及早衰的影响

 2006-2007年,采用4个转CpTI基因棉花材料及其受体研究了转CpTI基因对棉花苗病、黄萎病、后期叶斑病及早衰的影响。结果表明：3个转CpTI基因材料苗病死苗率均低于其对应的受体材料。在6月底和7月初黄萎病的发生初期,转基因材料黄萎病发生较受体材料轻,7月中旬到8月底9月初,不同品种以及同一品种的不同生育期之间差异较大,转基因材料C连续两年病指极显著低于对照受体材料。与非转基因材料相比,转CpTI基因材料叶斑病的发生显著加重,早衰发生也加重。     

不同抗性品种对棉花黄萎病菌致病力的影响

 以1个海岛棉品种和15个陆地棉品种为分离寄主,采取单孢分离,共分离出67个黄萎病菌系。以抗性差异较大的海岛棉品种Pima90-53和陆地棉品种冀棉20、邯208和中棉所8号为鉴别寄主,采用苗期营养钵定量注菌液法对分离菌系进行了致病力鉴定。结果表明,同一地块田间不同品种所分离菌系的致病力类型并非单一类型,而是由致病力连续变化的多种致病力菌系类型组成;由不同品种分离出的菌系致病力存在差异,寄主品种的抗性与分离的黄萎病菌系的致病力无关,但不同基因型品种可能会影响不同致病力菌落的消长。     

棉花GhSKIP35基因的克隆与表达分析

以中植棉KV1 SSH文库为基础,从陆地棉中植棉KV1中克隆出来泛素途径SKIP35基因,命名为GhSKIP35。该基因的开放阅读框全长为1863个核苷酸,编码620个氨基酸,含有1个ANK保守结构域。进化分析表明,Gh SKIP35蛋白与甜橙(Citrus sinensis)的SKIP35蛋白相似性最高。q RT-PCR分析显示:在接种大丽轮枝菌V991菌系后初期Gh SKIP35基因表达量激增,12 h便可达到顶峰,24~48 h逐渐下降,推测它在陆地棉抗黄萎病早期防卫反应中起重要作用。利用VIGS(Virus induced gene silencing)技术,在中植棉KV1中成功地沉默Gh SKIP35基因;对沉默棉株接种大丽轮枝菌V991鉴定显示,其病情指数为50.75,而野生型的中植棉KV1和转化空载体棉株的病情指数分别为9.38和11.54。Gh SKIP35基因沉默后,中植棉KV1对黄萎病的抗性丧失,表明Gh SKIP35基因在陆地棉抗黄萎病的过程中起重要作用。     

Identification of Brassica oleracea germplasm with improved resistance to Verticillium wilt

Two hundred and ninety‐nine accessions representing 11 cultivar groups of Brassica oleracea and eight additional accessions of the wild species B. cretica, B. incana, B. insularis and B. villosa were screened for resistance to Verticillium wilt. A disease index (DI) was calculated for each accession, and a correction of the DI was carried out to compensate for a fluctuating infection level between 11 independent trials. A total of 235 , or 77% of the accessions tested, had a DI_corr less or equal to the oilseed rape cv. ‘Express’ (DI_corr= 2.81), the reference cultivar. Only one accession of the wild species, B. incana, showed an enhanced level of resistance (DI_corr= 2.01). Twenty‐four accessions, distributed over eight cultivar groups of B. oleracea were selected for subsequent crosses involving B. rapa. Hybrid plants with 14 accessions were resultant and seed was obtained from crosses where the cultivar groups acephala, alboglabra, botrytis, capitata, gemmifera, italica and sabellica were used as female parents. When progeny of the produced resynthesized rapeseed lines were evaluated for Verticillium wilt resistance, three lines showed a significantly lower disease index (P ≥ 0.01) compared with the cv. ‘Express’. This source of resistance is now being crossed to advanced breeding material of oilseed rape.     

棉花黄萎病抗性与过氧化物酶同工酶分析

采用等电聚焦电泳方法 ,对棉花二对抗黄萎病近等基因系以及三对色素腺体近等基因系的子叶进行过氧化物同工酶 (POD)分析。结果表明 ,色素腺体和棉酚含量与棉花抗性没有必然的联系 ;环境条件对棉花抗性表现有一定的影响 ,但对子叶中 POD同工酶谱带无显著的影响 ;4条 POD同工酶酶带 (P4、P6、P7、P10 )的表现与棉花抗黄萎病性有关 ,这些酶带的特殊表现可用于棉花抗病性辅助鉴定和辅助育种