课题基金基金详情
大麦5HS染色体上叶锈病成株抗性基因(LR-APR)的精细定位与克隆
结题报告
批准号:
31201212
项目类别:
青年科学基金项目
资助金额:
25.0 万元
负责人:
徐延浩
依托单位:
学科分类:
C1307.作物基因组及遗传学
结题年份:
2015
批准年份:
2012
项目状态:
已结题
项目参与者:
杨飞、郭晓华、孙正祥、张文英、姚仪敏、付义川
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中文摘要
叶锈病是大麦的重要病害,利用成株抗性培育具持久抗性的大麦品种是控制叶锈病的根本措施。LR-APR是大麦5HS染色体上成株抗性QTL位点的主效基因,在WABAR2482×Pompadour F2群体中被解析为显性单基因,定位在2.0 cM区间内。在此基础上,本研究拟用大麦基因组测序信息,加密LR-APR基因区间分子标记,在上述群体中对该基因进行精细定位,筛选该基因共分离标记,比对5H染色体序列,构建该基因物理图谱,预测候选基因,比较候选基因亲本单体型及表达差异,缩小候选基因范围。构建抗性亲本的TAC文库,利用候选基因特异分子标记筛选TAC文库,对阳性克隆测序并转化感病品种。比较分析LR-APR基因位点在抗性品种中的多态性,明确5HS上是否存在成株抗性的多基因家族或LR-APR复等位基因。LR-APR基因的精细定位和克隆,将促进对大麦叶锈病成株抗性机制的理解,为大麦抗叶锈病育种提供信息。
英文摘要
Leaf rust caused by the fungus Puccinia hordei is one of the most significant diseases of barley worldwide. Adult plant resistance (APR) is considered potentially more durable for controlling barley leaf rust than seedling Rph (Resistance to Puccinia hordei) genes. The most preferable and economical method for controlling barley leaf rust is to develop and utilize the durable host resistance. LR-APR is the major gene of QTL on barley chromosome 5HS controlling adult plant resistance to Puccinia hordei, which was dissected into a dominant gene in F2 population of WABAR2482 × Pompadour and was mapped in a 2.0 cM interval (with 0.7 cM from marker bPb-0837 and 1.3 cM from marker EBmag0833). In this study, the barley genome sequence will be used to develop new markers in the LR-APR gene interval. The F2 population with 2600 individuals from WABAR2482 × Pompadour will be used to construct a fine mapping of LR-APR gene and to identify markers cosegregated with LR-APR. The cosegregated markers will be used to screen the sequences from barley 5HS chromosome to construct the physical map of LR-APR gene. The candidate genes will be predicted by bioinformatics approach. The haplotype and the expression patterns of the candidate genes will be analyzed in the DH population of Pompadour × Stirling to narrow the scope of candidate genes. The TAC library of resistance cultivar Pompadour will be constructed. The cosegregated markers will be used to screen the TAC library of Pompadour, the positive clone will be sequenced and transformed into susceptible barley cultivar. The polymorphism of LR-APR gene will be analyzed in different resistance cultivars to understand if there is a multiple gene family or multiple alleles of LR-APR gene for the 5HS locus resistance. The fine mapping and clone of LR-APR gene will improve the understanding of the molecular mechanism for barley leaf rust resistance and provide reference gene and molecular markers for barley resistance breeding.
叶锈病是大麦的重要病害,利用成株抗性培育具持久抗性的大麦品种是控制叶锈病的根本措施。最近的研究表明,大麦5HS染色体上可能存在一个叶锈病的抗病基因簇。利用不同来源的抗性材料构建的4个DH系:Baudin x RAH1995;Stirling x Baronesse;WI3407 x Baudin;Pompadour x Stirling进行多年多点的表型鉴定并结合分子标记技术进行基因定位。结果表明,5HS染色体上确实存在一个叶锈病的抗病基因簇。利用抗性品种WI3407与抗性品种Pompadour,Vada和RAH1995分别构建F2群体,抗性鉴定表明这些F2群体中都出现了感病单株,遗传分析表明,5HS染色体上至少有2个叶锈病的抗病(LR-APR基因)。在前期工作的基础上,构建了WABAR2482 X Pompadour的F2群体,并进行表型鉴定。利用2B-RAD技术开发分子标记,筛选出75位于基因区间内的分子标记,并进行重组单株的筛选和F2:3家系的表型验证。Pompadour 5HS染色体上LR-APR基因被定位在0.6 cM的区间内,并与标记MC463750紧密连锁。使用合作伙伴开发的大麦表型和基因型集成生物信息系统(Phenotype and Genome Integration System, PGIS)对LR-APR基因候选基因进行鉴定,初步确定了2个候选基因。
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DOI:10.1016/s2095-3119(13)60351-2
发表时间:2013-10
期刊:Journal of Integrative Agriculture
影响因子:4.8
作者:R. Lü;Yanhao Xu;Rodger Boyd;Xiao-qi Zhang;S. Broughton;Michael G. K. Jones;C. Li;Yaofeng Chen
通讯作者:R. Lü;Yanhao Xu;Rodger Boyd;Xiao-qi Zhang;S. Broughton;Michael G. K. Jones;C. Li;Yaofeng Chen
国内基金
海外基金