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利用从混合分离群体RNA测序中鉴定出的SNP标记对大白菜根肿病抗性基因进行精细定位

Fine Mapping of a Clubroot Resistance Gene in Chinese Cabbage Using SNP Markers Identified from Bulked Segregant RNA Sequencing.

作者信息

Huang Zhen, Peng Gary, Liu Xunjia, Deora Abhinandan, Falk Kevin C, Gossen Bruce D, McDonald Mary R, Yu Fengqun

机构信息

Saskatoon Research and Development Centre, Agriculture and Agri-Food Canada, SaskatoonSK, Canada.

State Key Laboratory of Crop Stress Biology for Arid Areas, College of Agronomy, Northwest A&F UniversityYangling, China.

出版信息

Front Plant Sci. 2017 Aug 28;8:1448. doi: 10.3389/fpls.2017.01448. eCollection 2017.

Abstract

Clubroot, caused by , is an important disease of canola () in western Canada and worldwide. In this study, a clubroot resistance gene () was identified and fine mapped in Chinese cabbage cv. "Jazz" using single-nucleotide polymorphisms (SNP) markers identified from bulked segregant RNA sequencing (BSR-Seq) and molecular markers were developed for use in marker assisted selection. In total, 203.9 million raw reads were generated from one pooled resistant (R) and one pooled susceptible (S) sample, and >173,000 polymorphic SNP sites were identified between the R and S samples. One significant peak was observed between 22 and 26 Mb of chromosome A03, which had been predicted by BSR-Seq to contain the causal gene . There were 490 polymorphic SNP sites identified in the region. A segregating population consisting of 675 plants was analyzed with 15 SNP sites in the region using the Kompetitive Allele Specific PCR method, and was fine mapped between two SNP markers, SNP_A03_32 and SNP_A03_67 with 0.1 and 0.3 cM from , respectively. Five SNP markers co-segregated with in this region. Variants were identified in 14 of 36 genes annotated in the target region. The numbers of poly variants differed among the genes. Four genes encode TIR-NBS-LRR proteins and two of them and , had high numbers of polymorphic variants and so are the most likely candidates of .

摘要

根肿病由[病原体名称未给出]引起,是加拿大西部及全球范围内油菜([油菜品种未给出])的一种重要病害。在本研究中,在中国白菜品种“Jazz”中鉴定并精细定位了一个根肿病抗性基因([基因名称未给出]),使用从混合分离群体RNA测序(BSR-Seq)中鉴定出的单核苷酸多态性(SNP)标记,并开发了分子标记用于标记辅助选择。总共从一个混合抗性(R)样本和一个混合感病(S)样本中产生了2.039亿条原始读数,并且在R和S样本之间鉴定出超过17.3万个多态性SNP位点。在A03染色体的22至26 Mb之间观察到一个显著峰值,BSR-Seq预测该区域包含致病基因[基因名称未给出]。在该区域鉴定出490个多态性SNP位点。使用竞争性等位基因特异性PCR方法,对由675株植物组成的分离群体进行该区域的15个SNP位点分析,[基因名称未给出]被精细定位在两个SNP标记SNP_A03_32和SNP_A03_67之间,分别距[基因名称未给出]0.1和0.3 cM。在该区域有5个SNP标记与[基因名称未给出]共分离。在[目标区域未明确指出]注释的36个基因中的14个中鉴定出变异。多态变异的数量在不同基因之间有所不同。四个基因编码TIR-NBS-LRR蛋白,其中两个基因[基因名称未给出]和[基因名称未给出]具有大量的多态变异,因此是[基因名称未给出]最有可能的候选基因。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e323/5581393/b626b403efac/fpls-08-01448-g001.jpg

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