小麥品種Madsen抗禾谷孢囊線蟲的遺傳控制與抗性利用
發(fā)布時間：2018-10-31 13:05
【摘要】：禾谷孢囊線蟲(Cereal cyst nematode,CCN)已報道在我國16個省(市、自治區(qū))的小麥生產(chǎn)地區(qū)發(fā)生危害,地處黃淮冬麥區(qū)的河南省受害面積和程度尤其嚴重,對小麥的安全生產(chǎn)構(gòu)成嚴重威脅。Heteroder avenae和H.filipjevi兩種禾谷孢囊線蟲在我國混合發(fā)生且致病型多樣,增加了防治的難度。當前我國的絕大多數(shù)品種對兩種禾谷孢囊線蟲都不具有抗性,而且有效抗源也匱乏,嚴重影響了利用寄主抗性有效防治禾谷孢囊線蟲。研究發(fā)現(xiàn)美國小麥品種Madsen具有良好的線蟲抗性。本研究圍繞Madsen我國發(fā)生的禾谷孢囊線蟲抗性、對小麥根際土壤線蟲群體的影響、抗兩種禾谷孢囊線蟲的遺傳控制機制和抗性種質(zhì)創(chuàng)新等進行研究,主要結(jié)果如下:1.通過多年重復(fù)的田間病圃鑒定和溫室接種鑒定,Madsen對來自河南、山東和安徽等地的4個H.filipjevi群體和6個H.avenae群體都表現(xiàn)抗病或中抗反應(yīng)型,根系孢囊數(shù)均顯著少于感病對照溫麥19,表明Madsen對我國多個地區(qū)禾谷孢囊線蟲群體具有穩(wěn)定抗性和廣譜抗性。2.在PF-127膠體中,Madsen和溫麥19的根尖對H.aavenae都表現(xiàn)出吸引性,但在禾谷孢囊線蟲危害地塊,Madsen根系正常生長,溫室接種條件下早期(25天)侵入Madsen根系的線蟲數(shù)目少且最終形成孢囊少。所以,Madsen的抗性機制可能是抑制線蟲的侵入或使侵入的線蟲無法完成生活史。連續(xù)兩年分別在河南H.filipjevi和H.avenae病田,采用人工計數(shù)白雌蟲、繁殖系數(shù)法(Rf)和南澳大利亞研究與開發(fā)研究所的PreDicta B分子檢測方法對Madsen等7個不同抗性品種根際土壤線蟲群體進行分析,三種評價方法分析結(jié)果一致表明,種植Madsen等抗性品種能夠減少線蟲在當季小麥根系上增殖,同時降低下一生長季節(jié)根際土壤中線蟲的密度。3.基于187個Madsen×良星99 F6:9重組近交系(RIL)為作圖群體,采用IlluminaiSelect 90K SNP芯片結(jié)合SSR標記,利用3219個多態(tài)性標記構(gòu)建了覆蓋小麥A、B和D基因組21條染色體的高密度遺傳連鎖圖譜,包含2466個位點,覆蓋染色體總長度為4576.11 cM。根據(jù)3年的田間病圃(2013-2015年)和2年(2013-2014年)溫室接種共5個環(huán)境對H.filipjevi.線蟲的表型鑒定數(shù)據(jù),進行抗線蟲QTL分析,在7D染色體上發(fā)現(xiàn)一個主效QTL位點QCre-ma7D,解釋表型變異率為13.6%-42.0%,7D染色體上未報道有抗H.filipjevi的基因或QTL,因此,QCre-ma7D可能是抗H.filipjevi新的主效QTL位點。在H.avenae溫室接種和田間病圃2個環(huán)境中檢測到一個位于2A染色體的QTL位點QCre-ma2A,解釋表型變異率為15.4%-19.6%,該QTL位點可能是已知抗線蟲基因Cre5。獲得與QTL抗性位點緊密連鎖的SNP標記,將與QCre-ma7D緊密連鎖的三個SNP(Kukri-_c45628_892,BS00021745_51 和Kukri_rep_c68335_607)轉(zhuǎn)化為分型結(jié)果清晰的KASP標記,建立了QCre-ma7D的分子標記輔助選擇技術(shù)。根據(jù)對Madsen系譜涉及品種的表型鑒定結(jié)果結(jié)合分子標記檢測結(jié)果,Madsen對Hfilipjevi和H.atvenae的抗性可能來自親本VPM1攜帶的偏凸山羊草染色體片段。4.Madsen抗兩種禾谷孢囊線蟲但極晚熟,且不抗白粉病,影響其抗性利用,而良星99攜帶抗白粉病基因Pm52。利用分子標記輔助選擇技術(shù),從Madsen×良星99雜交組合中選育出兼抗兩種線蟲和白粉病的ML99-18和ML99-46,兩個家系遺傳了來自親本Madsen的抗線蟲QTL和良星99的抗白粉病基因Pm52,而且抽穗期和農(nóng)藝性狀都與親本良星99相近,可作為抗病蟲新種質(zhì)資源加以利用。
[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.
【學位授予單位】：中國農(nóng)業(yè)大學
【學位級別】：博士
【學位授予年份】：2017
【分類號】：S435.12
本文編號：2302295
[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.
【學位授予單位】：中國農(nóng)業(yè)大學
【學位級別】：博士
【學位授予年份】：2017
【分類號】：S435.12
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