首页 > 硕士 > 农学 > 正文

水稻纹枯病抗性遗传分析及稻种资源的抗病鉴定与评价

Genetic Analysis of Rice Sheath Blight Resistance and Rice Germplasm Identification and Evaluation of Resistance

作者: 专业:作物遗传育种 导师:罗利军 年度:2010 学位:硕士  院校: 华中农业大学

Keywords

Rice sheath blight, Recombinant inbred lines(RILs), Quantitative traitlocis(QTLs), Identification and evaluation

        由立枯丝核菌(Rhizoctonia solani Kuhn)引起的水稻纹枯病是最为严重的真菌性水稻病害之一,对水稻的产量和品质都有很大的影响。本研究利用155个株系组成的沪旱1B/RSB02重组自交系群体,构建了一张包含163个SSR标记的水稻分子遗传连锁图谱,全长2588.8cM,标记间的平均间距为15.88cM。采用短牙签嵌入接种方法,分别在三个环境条件下对该群体的病级(disease rate, DR)、病斑长度(lesion length, LL)、病斑高度(lesion heigth, LH)、相对病斑长(relative lesion length, RLL)、相对病斑高(relative lesion heigth, RLH)、抽穗期(heading date, HD)、株高(plant height,PH)、结实率(spikelet fertility percentage, SF)和千粒重(1000-grain weight, TGW)9个性状进行了QTL定位,并分析了环境对水稻纹枯病抗性的影响。采用自然诱病和人工牙签接种法对中国水稻微核心种质和“全球水稻分子育种计划”亲本进行了多年多点的纹枯病抗性鉴定和评价。主要研究结果如下:1.2008年在上海(E1)、2008年在海南(E2)和2009年在上海(E3)对亲本和群体材料进行了纹枯病抗性鉴定和QTL定位分析。在三个环境中,共检测到22、22和20个主效QTLs,20、23和34对上位性QTLs控制DR、LL、LH、RLL、RLH、PH、HD、SF和TGW性状。在E1和E2环境中都检测到位于第1染色体RM3442-RM5501区间的QTL同时影响着DR、PH、RLL、RLH,位于第6染色体RM1369-RM8121区间的QTL影响着RLL;在E1和E3环境中都检测到位于第1染色体RM3233-RM8134区间的QTL对RLH的影响。这些重复定位的QTL的抗性等位基因都来自抗病亲本RSB02,其中位于第1染色体RM3442-RM5501区间的QTL在E1和E2环境中分别解释了病级表型变异的30.19%和5.63%,在增强纹枯病抗性的同时也使株高大幅度增加。2.利用三个环境条件下的表型数据进行联合QTLs定位,检测到35个主效QTLs,分别位于第1,2,3,4,5,6,7,9染色体上,多数位点的抗性等位基因来自抗病亲本RSB02,位于第1染色体RM3442-RM5501区间和RM3233-RM8134区间的QTLs同时控制着DR、PH、RLL、RLH。检测到46对上位性QTLs,其中只有HD没有检测到与环境互作的上位性QTLs,其它性状均有与环境存在互作的QTLs.3.通过相关性分析,在三个环境条件下DR与LL、LH、RLL和RLH均表现为极显著正相关,而与PH,SF和TGW呈负相关,其中与PH的负相关性达极显著水平。平均相关系数最高的是DR与RLH达到了0.913,其次是与RLL的平均相关系数也达到0.778。在联合定位中检测到的控制DR的6个QTLs中,5个QTLs被检测到同时控制RLL和RLH,2个与控制PH的QTLs位于同一染色体区间内。本研究结果表明,株高对水稻纹枯病抗性影响较大。RLH和RLL与DR极显著相关,且操作简单易行,受人为主观因素的影响较小,可作为水稻纹枯病抗性鉴定的重要评价指标之一4.在2008年恩施和海南分别采取自然诱病和人工接种方法对159份中国水稻微核心种质和137份“全球水稻分子育种计划”亲本进行抗纹枯病大田鉴定和评价,重复鉴定材料的抗性级别以发病重的为准。发现抗病品种有4份,中抗品种有68份,分别占鉴定总数的1.4%和23%,其中有14份材料在两地鉴定中对纹枯病均表现为中抗水平;中感、感病和高感品种分别占33.8%、22%和19.9%。抽穗期、剑叶枕高、株高和剑叶枕高/株高的比值与病级之间呈极显著负相关,病斑长、病斑高、病斑长/剑叶枕高的比值、病斑长/株高的比值、病斑高/剑叶枕高的比值和病斑高/株高的比值与病级之间呈极显著正相关。
    Rice sheath blight, caused by Rhizoctonia solani, is one of the most prevalent fungal diseases in rice, which leads to a great loss of rice yield and quality worldwide. In the present study, a recombinant inbred lines (RILs) population which consists of 155 individuals was derived from a cross between HuHan 1B and RSB02, and a genetic linkage map was constructed based on the RILs and 163 SSR markers, which covered a total of 2588.8cM with an average interval of 15.88cM between adjacent markers. The QTLs analysis and the effect of environment for 9 traits on resistance to rice sheath blight, including disease rate (DR), lesion length (LL), lesion height (LH), relative lesion length (RLL), relative lesion height (RLH), heading date (HD), plant height(PH), spikelet fertility percentage (SF),1000-grain weight (TGW), were conducted according to the phenotypic data across three environments by using the toothpick inoculation method. Cultivars from China rice mini core collection and "global rice molecular breeding program" were identified and evaluated for resistance to rice sheath blight under natural induction or artificial inoculation. Main results are as following:1. Rice sheath blight resistance of parents and RILs population were identified in Shanghai in 2008(E1), Hainan (E2) and Shanghai in 2009 (E3). A total of 22,22 and 20 main QTLs,20,23 and 34 epistatic QTLs were detected across three environments, respectively, for nine traits, DR, LL, LH, RLL, RLH, PH, HD, SF and TGW. Among them, the QTL that influenced DR, PH, RLL and RLH simultaneously locating between RM3442 and RM5501 on chromosome 1 and the QTL for RLL locating at interval of RM1369-RM8121 on chromosome 6 were identified repeatedly in E1 and E2. While, the QTL for RLH flanked by RM3233 andRM8134 on chromosome 1 was identified repeatedly in E1 and E3. The resistance alleles of these QTLs came from resistant parent, RSB02. The QTL that located at interval of RM3442-RM5501 on chromosome 1 enhanced resistance to sheath blight, while greatly increased the plant height.2. Combined QTLs analysis was conducted with the total data in three environments. 35 main QTLs were detected which located on chromosome 1,2,3,4,5,6,7,9. The resistance alleles of most QTLs came from resistant parent RSB02. The QTLs at interval of RM3442-RM5501 and RM3233-RM8134 on chromosome 1 influenced DR, PH, RLL and RLH. And 46 epistatic QTLs were detected for nine traits. All traits had QTLs with significant Q X E interation excepted HD.3. Correlation analysis shoewed that DR had significant positive correlations with LL, LH, RLL, and RLH but negatively correlated with PH, SF and TGW. The average correlation coefficients between DR and RLH, RLL were 0.913 and 0.778, respectively. Among 6 QTLs for DR detected by combined QTLs analysis,5 QTLs were identified to control RLL and RLH and 2 QTLs to influence PH simultaneously. The results showed that plant height had great impact on rice sheath blight resistance. RLH and RLL could be used as important indices indicading sheath blight resistance due to its high correlation with DR and simple, objective measurement.4.159 rice cultivars from China mini core collection and 137 varieties from the "global rice molecular breeding program" were identified and evaluated for resistance to rice sheath blight using naturally induced and artificial inoculation methods in Enshi and Shanghai in 2008. The number of resistant and moderately resistant germplasm were 4 and 68. The percentage of resistant, moderately resistant, moderately susceptible, susceptible and highly susceptible germplasm were 1.4%,23%,33.8%,22%,19.9%, respectively. HD, FLPH, PH, the ratio of FLPH and PH were significant negatively correlated with DR. LL, LH, the ratio of LL and FLPH, the ratio of LL and PH, the ratio of LH and FLPH and the ratio of LH and PH were significant positively correlated with DR.
        

水稻纹枯病抗性遗传分析及稻种资源的抗病鉴定与评价

摘要6-8
Abstract8-9
缩略名词表10-11
1 文献综述11-20
    1.1 水稻纹枯病病原菌生物学特性11
    1.2 水稻纹枯病的田间抗性鉴定体系11-13
        1.2.1 接种方法11-12
        1.2.2 接种时期和调查时期12
        1.2.3 病情评价标准和指标12-13
    1.3 水稻纹枯病抗源的筛选和抗性种质创制13-15
    1.4 QTL定位和分子标记辅助选择15-17
        1.4.1 QTL定位的方法15-16
        1.4.2 QTL定位的群体16
        1.4.3 分子标记辅助选择16-17
    1.5 水稻纹枯病抗性的遗传研究17-20
    1.6 本研究的意义和目的20
2 利用重组自交系分析水稻纹枯病抗性位点和效应20-52
    2.1 材料和方法20-23
        2.1.1 试验材料20-21
        2.1.2 试验方法21-22
            2.1.2.1 田间试验设计21
            2.1.2.2 水稻纹枯病菌接种21
            2.1.2.3 病情评价标准21-22
            2.1.2.4 性状调查22
        2.1.3 表型数据统计分析22
        2.1.4 亲本多态性检测、群体基因型分析和遗传图谱构建22-23
            2.1.4.1 DNA提取22
            2.1.4.2 亲本多态性筛选22-23
            2.1.4.3 SSR分析23
            2.1.4.4 遗传连锁图谱的构建和QTL分析23
    2.2 结果和分析23-38
        2.2.1 亲本与群体的纹枯病抗性表现23-24
        2.2.2 接种纹枯病对产量相关性状的影响24
        2.2.3 RILs群体产量相关性状的t测验24-28
        2.2.4 RILs群体纹枯病抗性相关性状的相关性分析28-29
        2.2.5 连锁遗传图谱构建和群体基因型分析29-30
        2.2.6 单个环境下的水稻纹枯病抗性QTLs分析30-36
            2.2.6.1 E1中水稻纹枯病抗性QTLs分析30-32
            2.2.6.2 E2中水稻纹枯病抗性QTLs分析32-33
            2.2.6.3 E3中水稻纹枯病抗性QTLs分析33-35
            2.2.6.4 单个环境条件下水稻纹枯病抗性相关QTLs结果的比较35-36
        2.2.7 水稻纹枯病抗性相关三个环境下的联合QTLs定位36-38
            2.2.7.1 水稻纹枯病抗性相关QTLs及其与环境的互作36-37
            2.2.7.2 水稻纹枯病抗性相关上位性互作位点及其与环境的互作37-38
    2.3 讨论38-52
        2.3.1 抗纹枯病QTL的单季定位结果与联合定位结果比较38
        2.3.2 QTLs定位结果的比较38-39
        2.3.3 环境对纹枯病抗性的影响39-40
        2.3.4 水稻农艺性状对纹枯病抗性鉴定的影响40-41
        2.3.5 水稻纹枯病发病程度的评价指标41-52
3 水稻优异种质资源的纹枯病抗性鉴定与评价52-60
    3.1 材料和方法52-53
        3.1.1 材料52
        3.1.2 方法52
        3.1.3 病情调查52
        3.1.4 抗病性评价52-53
        3.1.5 农艺性状和病级的相关性及差异显著性分析53
    3.2 结果和分析53-57
        3.2.1 纹枯病抗性鉴定结果53
        3.2.2 纹枯病抗性鉴定稳定性的分析53-54
        3.2.4 中国水稻微核心种质的纹枯病抗性表现54-55
        3.2.5 全球水稻分子育种亲本的纹枯病抗性表现55
        3.2.6 农艺性状和病级的相关性及差异显著性分析55-57
    3.3 讨论57-60
        3.3.1 抗纹枯病稻种资源57-58
        3.3.2 水稻纹枯病抗性影响因素58
        3.3.3 纹枯病抗性鉴定指标58-60
参考文献60-65
致谢65-66
附录66-69
    附录Ⅰ 植物DNA提取步骤66-67
        附录Ⅰ-1 大样法(改良CTAB法)66
        附录Ⅰ-2 小样法66-67
        附录Ⅰ-3 小样改进法67
    附录Ⅱ SSR反应体系和扩增67
    附录Ⅲ 电泳检测67-69
        附录Ⅲ-1 琼脂糖电泳检测67-68
        附录Ⅲ-2 PAGE电泳检测68-69
        下载全文需50


本文地址:

上一篇:旱稻IRAT109辐照突变体的抗旱性鉴定和窄叶突变体(nal8)的初步定位
下一篇:鹅CYP7A1基因克隆、表达调节及表达规律的研究

分享到: 分享水稻纹枯病抗性遗传分析及稻种资源的抗病鉴定与评价到腾讯微博           收藏
评论排行
公告