小麦品种Madsen抗禾谷孢囊线虫的遗传控制与抗性利用

发布时间：2018-10-31 13:05

【摘要】：禾谷孢囊线虫(Cereal cyst nematode,CCN)已报道在我国16个省(市、自治区)的小麦生产地区发生危害,地处黄淮冬麦区的河南省受害面积和程度尤其严重,对小麦的安全生产构成严重威胁。Heteroder avenae和H.filipjevi两种禾谷孢囊线虫在我国混合发生且致病型多样,增加了防治的难度。当前我国的绝大多数品种对两种禾谷孢囊线虫都不具有抗性,而且有效抗源也匮乏,严重影响了利用寄主抗性有效防治禾谷孢囊线虫。研究发现美国小麦品种Madsen具有良好的线虫抗性。本研究围绕Madsen我国发生的禾谷孢囊线虫抗性、对小麦根际土壤线虫群体的影响、抗两种禾谷孢囊线虫的遗传控制机制和抗性种质创新等进行研究,主要结果如下:1.通过多年重复的田间病圃鉴定和温室接种鉴定,Madsen对来自河南、山东和安徽等地的4个H.filipjevi群体和6个H.avenae群体都表现抗病或中抗反应型,根系孢囊数均显著少于感病对照温麦19,表明Madsen对我国多个地区禾谷孢囊线虫群体具有稳定抗性和广谱抗性。2.在PF-127胶体中,Madsen和温麦19的根尖对H.aavenae都表现出吸引性,但在禾谷孢囊线虫危害地块,Madsen根系正常生长,温室接种条件下早期(25天)侵入Madsen根系的线虫数目少且最终形成孢囊少。所以,Madsen的抗性机制可能是抑制线虫的侵入或使侵入的线虫无法完成生活史。连续两年分别在河南H.filipjevi和H.avenae病田,采用人工计数白雌虫、繁殖系数法(Rf)和南澳大利亚研究与开发研究所的PreDicta B分子检测方法对Madsen等7个不同抗性品种根际土壤线虫群体进行分析,三种评价方法分析结果一致表明,种植Madsen等抗性品种能够减少线虫在当季小麦根系上增殖,同时降低下一生长季节根际土壤中线虫的密度。3.基于187个Madsen×良星99 F6:9重组近交系(RIL)为作图群体,采用IlluminaiSelect 90K SNP芯片结合SSR标记,利用3219个多态性标记构建了覆盖小麦A、B和D基因组21条染色体的高密度遗传连锁图谱,包含2466个位点,覆盖染色体总长度为4576.11 cM。根据3年的田间病圃(2013-2015年)和2年(2013-2014年)温室接种共5个环境对H.filipjevi.线虫的表型鉴定数据,进行抗线虫QTL分析,在7D染色体上发现一个主效QTL位点QCre-ma7D,解释表型变异率为13.6%-42.0%,7D染色体上未报道有抗H.filipjevi的基因或QTL,因此,QCre-ma7D可能是抗H.filipjevi新的主效QTL位点。在H.avenae温室接种和田间病圃2个环境中检测到一个位于2A染色体的QTL位点QCre-ma2A,解释表型变异率为15.4%-19.6%,该QTL位点可能是已知抗线虫基因Cre5。获得与QTL抗性位点紧密连锁的SNP标记,将与QCre-ma7D紧密连锁的三个SNP(Kukri-_c45628_892,BS00021745_51 和Kukri_rep_c68335_607)转化为分型结果清晰的KASP标记,建立了QCre-ma7D的分子标记辅助选择技术。根据对Madsen系谱涉及品种的表型鉴定结果结合分子标记检测结果,Madsen对Hfilipjevi和H.atvenae的抗性可能来自亲本VPM1携带的偏凸山羊草染色体片段。4.Madsen抗两种禾谷孢囊线虫但极晚熟,且不抗白粉病,影响其抗性利用,而良星99携带抗白粉病基因Pm52。利用分子标记辅助选择技术,从Madsen×良星99杂交组合中选育出兼抗两种线虫和白粉病的ML99-18和ML99-46,两个家系遗传了来自亲本Madsen的抗线虫QTL和良星99的抗白粉病基因Pm52,而且抽穗期和农艺性状都与亲本良星99相近,可作为抗病虫新种质资源加以利用。
[Abstract]:Cereal cyyssal nematode (CCN) has been reported to be harmful to the wheat production area of 16 provinces (cities, autonomous regions) in China. Howeder avenae and H. filipjevi are mixed in China and have a variety of pathogenic types, increasing the difficulty of prevention and treatment. Most of the varieties in China do not have resistance to two kinds of cereal-type nematodes, and there is a shortage of effective anti-sources, which seriously affects the prevention and treatment of cereal-parasitic nematodes by using host resistance. The study found that the American wheat variety Madsen had good nematode resistance. The main results are as follows: 1. Through multi-year repeated field disease nursery identification and greenhouse inoculation identification, Madsen showed that four H. filipjevi populations and 6 H. aviae groups from Henan, Shandong and Anhui provinces showed disease-resistant or anti-reactive type, and the root nodule count was significantly less than that of control temperature wheat 19. It is shown that Madsen has stable resistance and chemotaxis to the nematode population in various regions of China. In PF-127 colloid, the root tips of Madsen and Wenmai 19 showed the attraction of H. aavenae, but the number of nematodes invaded by Madsen root system was less and the root formation was less than that of Madsen root system at the same time (25 days). Therefore, Madsen's anti-sexual mechanism may be the inhibition of nematode invasion or the inability of invasive nematodes to complete the disease. Seven different resistant cultivars of Madsen et al. were analyzed by means of artificial counts of white female worm, propagation coefficient method (Rf) and the Prepta B molecular detection method of South Australia Research and Development Institute in Henan H. filipjevi and H. avenae disease fields for two consecutive years. The results of the three evaluation methods showed that the resistant varieties such as Madsen were able to reduce the proliferation of nematodes on the root of the quaternary wheat, while reducing the density of nematodes in the next growing season. A high density genetic linkage map covering 21 chromosomes of wheat A, B and D was constructed based on 187 Madsen's Wisconsin 99 F6: 9 recombinant inbred line (RIL) as a mapping population, and a high density genetic linkage map covering 21 chromosomes of wheat A, B and D was constructed using 3219 polymorphic markers. The total length of the chromosome covering the chromosome was 4576. 11%. H. filipjevi was inoculated in 3 years of field disease nursery (2013-2015) and 2 years (2013-2014) greenhouse. According to the phenotypic identification data of nematodes, a QTL analysis of nematode resistance was carried out, a major QTL site QCe-ma7D was found on the 7D chromosome, and the phenotypic variation rate was 13. 6% -42. 0%, and no gene or QTL against H. filipjevi was reported on the 7D chromosome. Therefore, QCre-ma7D may be a new major QTL site against H. filipjevi. A QTL locus QCe-ma2A, located on the 2A chromosome, was detected in two environments of H. avenae greenhouse inoculation and field disease nursery, and the phenotypic variation rate was 15. 4%-19. 6%, which could be a known anti-nematode gene Cre5. Three SNPs linked with QCre-ma7D (Kukri-_ c45628 _ 892, BS00021745 _ 51 and Kukri _ rep _ c68335 _ 607), which are closely linked to the QTL resistance locus, were transformed into KASP markers with clear typing results, and molecular marker-assisted selection techniques for QCre-ma7D were established. The resistance of Madsen to Hfilipjevi and H. atvenae may be derived from the chromosome segment of Leymus chinensis chromosome carried by the parent VPM1 according to the results of phenotypic identification and molecular marker detection of Madsen's pedigree. and the good star 99 carries the powdery mildew resistance gene Pm52. ML99-18 and ML99-46, which were both resistant to both nematodes and powdery mildew, were selected from the hybrid combination of Madsen and Liangliang Star 99 using molecular marker-assisted selection techniques, and two families inherited the anti-nematode QTL from the parent Madsen and the powdery mildew resistance gene Pm52 of Liangxing 99, and the heading date and the agronomic characters are similar to that of the parent good star 99, and can be used as a new germplasm resource of the disease-resistant insect.
【学位授予单位】：中国农业大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：S435.12

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