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通过结合 bulked segregant analysis 和 RNA-seq,快速鉴定玉米中烟嘧磺隆敏感性的候选基因(Nss)。

Rapid identification of a candidate nicosulfuron sensitivity gene (Nss) in maize (Zea mays L.) via combining bulked segregant analysis and RNA-seq.

机构信息

Institute of Cereal and Oil Crops, Hebei Academy of Agriculture and Forestry Sciences, Shijiazhuang, 050035, China.

Department of Crop, Soil and Environmental Science, Auburn University, Auburn, AL, 36849, USA.

出版信息

Theor Appl Genet. 2019 May;132(5):1351-1361. doi: 10.1007/s00122-019-03282-8. Epub 2019 Jan 16.

Abstract

A candidate nicosulfuron sensitivity gene Nss was identified by combining bulked segregant analysis and RNA-seq. Multiple mutations of this gene were discovered in nicosulfuron-sensitive maize compared with the tolerant. It has been demonstrated that variabilities exist in maize response to nicosulfuron. Two nicosulfuron-sensitive inbred lines (HB39, HB41) and two tolerant inbred lines (HB05, HB09) were identified via greenhouse and field trials. Genetic analysis indicated that the sensitivity to nicosulfuron in maize was controlled by a single, recessive gene. To precisely and rapidly map the nicosulfuron sensitivity gene (Nss), two independent F segregating populations, Population A (HB41 × HB09) and Population B (HB39 × HB05), were constructed. By applying bulked segregant RNA-Seq (BSR-Seq), the Nss gene was, respectively, mapped on the short arm of chromosome 5 (chr5: 1.1-15.3 Mb) and (chr5: 0.5-18.2 Mb) using two populations, with 14.2 Mb region in common. Further analysis revealed that there were 43 and 119 differentially expressed genes in the mapping intervals, with 18 genes in common. Gene annotation results showed that a cytochrome P450 gene (CYP81A9) appeared to be the candidate gene of Nss associated with nicosulfuron sensitivity in maize. Sequence analysis demonstrated that two common deletion mutations existed in the sensitive maize, which might lead to the nicosulfuron sensitivity in maize. The results will make valuable contributions to the understanding of molecular mechanism of herbicide sensitivity in maize.

摘要

通过结合 bulked segregant analysis 和 RNA-seq,鉴定出候选的烟嘧磺隆敏感基因 Nss。与耐受品种相比,在烟嘧磺隆敏感的玉米中发现了该基因的多个突变。已经证明玉米对烟嘧磺隆的反应存在变异性。通过温室和田间试验,鉴定出了两个烟嘧磺隆敏感的自交系(HB39、HB41)和两个耐受的自交系(HB05、HB09)。遗传分析表明,玉米对烟嘧磺隆的敏感性由一个单一的隐性基因控制。为了精确和快速定位烟嘧磺隆敏感基因(Nss),构建了两个独立的 F 分离群体,群体 A(HB41×HB09)和群体 B(HB39×HB05)。通过应用 bulked segregant RNA-Seq(BSR-Seq),分别在两个群体中将 Nss 基因定位在 5 号染色体的短臂上(chr5:1.1-15.3 Mb)和(chr5:0.5-18.2 Mb),共有 14.2 Mb 区域。进一步分析表明,在定位区间内有 43 个和 119 个差异表达基因,其中有 18 个基因是共同的。基因注释结果表明,一个细胞色素 P450 基因(CYP81A9)似乎是与玉米中烟嘧磺隆敏感性相关的 Nss 的候选基因。序列分析表明,在敏感玉米中存在两个共同的缺失突变,这可能导致玉米对烟嘧磺隆的敏感性。这些结果将有助于理解玉米中除草剂敏感性的分子机制。

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