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玉米(Zea mays L.)抗茎腐病基因Rfg1的遗传分析与分子定位

Genetic analysis and molecular mapping of maize (Zea mays L.) stalk rot resistant gene Rfg1.

作者信息

Yang D E, Zhang C L, Zhang D S, Jin D M, Weng M L, Chen S J, Nguyen H, Wang B

机构信息

Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, 100101 Beijing, China.

出版信息

Theor Appl Genet. 2004 Feb;108(4):706-11. doi: 10.1007/s00122-003-1466-y. Epub 2003 Nov 27.

Abstract

One single pathogen Fusarium graminearum Schw. was inoculated to maize inbred lines 1,145 (Resistant) and Y331 (Susceptive), and their progenies of F(1), F(2) and BC(1)F(1) populations. Field statistical data revealed that all of the F(1) individuals were resistant to the disease and that the ratio of resistant plants to susceptive plants was 3:1 in the F(2) population, and 1:1 in the BC(1)F(1 )population. The results revealed that a single dominant gene controls the resistance to F. graminearum Schw. The resistant gene to F. graminearum Schw. was denominated as Rfg1 according to the standard principle of the nomenclature of the plant disease resistant genes. RAPD (randomly amplified polymorphic DNA) combined with BSA (bulked segregant analysis) analysis was carried out in the developed F(2) and BC(1)F(1 )populations, respectively. Three RAPD products screened from the RAPD analysis with 820 Operon 10-mer primers showed the linkage relation with the resistant gene Rfg1. The three RAPD amplification products (OPD-20(1000), OPA-04(1100) and OPY-04(900)) were cloned and their copy numbers were determined. The results indicated that only OPY-04(900) was a single-copy sequence. Then, OPY-04(900) was used as a probe to map the Rfg1 gene with a RIL F(7) mapping population provided by Henry Nguyen, which was developed from the cross "S3xMo17". Rfg1 was primarily mapped on chromosome 6 between the two linked markers OPY-04(900) and umc21 (Bin 6.04-6.05). In order to confirm the primary mapping result, 25 SSR (simple sequence repeat) markers and six RFLP (restriction fragment length polymorphism) markers in the Rfg1 gene-encompassing region were selected, and their linkage relation with Rfg1 was analyzed in our F(2) population. Results indicated that SSR marker mmc0241 and RFLP marker bnl3.03 are flanking the Rfg1 gene with a genetic distance of 3.0 cM and 2.0 cM, respectively. This is the first time to name and to map a single resistant gene of maize stalk rot through a single pathogen inoculation and molecular marker analysis.

摘要

将单一病原菌禾谷镰刀菌接种到玉米自交系1145(抗病)和Y331(感病)及其F(1)、F(2)和BC(1)F(1)群体的后代中。田间统计数据显示,所有F(1)个体均抗病,F(2)群体中抗病植株与感病植株的比例为3:1,BC(1)F(1)群体中该比例为1:1。结果表明,一个显性单基因控制对禾谷镰刀菌的抗性。根据植物抗病基因命名的标准原则,将禾谷镰刀菌的抗性基因命名为Rfg1。分别在构建的F(2)和BC(1)F(1)群体中进行了RAPD(随机扩增多态性DNA)与BSA(混合群体分析法)分析。用820个Operon 10聚体引物进行RAPD分析筛选出的3个RAPD产物显示与抗性基因Rfg1存在连锁关系。对这3个RAPD扩增产物(OPD - 20(1000)、OPA - 04(1100)和OPY - 04(900))进行克隆并测定其拷贝数。结果表明,只有OPY - 04(900)是单拷贝序列。然后,以OPY - 04(900)为探针,利用Henry Nguyen提供的由“S3×Mo17”杂交构建的RIL F(7)作图群体对Rfg1基因进行定位。Rfg1初步定位在第6号染色体上,位于两个连锁标记OPY - 04(900)和umc21之间(Bin 6.04 - 6.05)。为了验证初步定位结果,在Rfg1基因所在区域选择了25个SSR(简单序列重复)标记和6个RFLP(限制性片段长度多态性)标记,并在我们的F(2)群体中分析它们与Rfg1的连锁关系。结果表明,SSR标记mmc0241和RFLP标记bnl3.03分别在Rfg1基因两侧,遗传距离分别为3.0 cM和2.0 cM。这是首次通过单一病原菌接种和分子标记分析对玉米茎腐病的单个抗性基因进行命名和定位。

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