Identification and fine mapping of a stay-green gene (Brnye1) in pakchoi (Brassica campestris L. ssp. chinensis)

Identification and fine mapping of a stay-green gene (Brnye1) in pakchoi (Brassica campestris L. ssp. chinensis)
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DOI:
10.1007/s00122-017-3028-8
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发表时间:
2018-03
影响因子:
5.4
通讯作者:
Nan Wang;Zhiyong Liu;Yun Zhang;Chengyu Li;H. Feng
Nan Wang;Zhiyong Liu;Yun Zhang;Chengyu Li;H. Feng
中科院分区:
农林科学1区
文献类型:
--
作者:
Nan Wang;Zhiyong Liu;Yun Zhang;Chengyu Li;H. Feng

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关键信息利用批量分离分析和下一代测序相结合的方法,我们确定了控制小白菜保绿特性的Brnye1基因。序列分析确定Bra019346为候选基因。摘要“保绿”是指叶片在衰老过程中保持绿色的一种植物性状。这一特性在小白菜(Brassica campestrisL.)的栽培中是有用的。中国),这是一种绿叶产品。本研究旨在找出影响小白菜保绿性状的基因。我们在小白菜中发现了一个保持绿色的突变体,我们称之为“奈伊”。遗传分析表明,保绿性状受1对隐性基因Brnye1控制。利用BSA-seq方法,在A03染色体上定位了一个3.0Mb的候选区域,通过连锁分析,将Brnye1定位在SSR标记SSRWN27和SSRWN30之间。我们在该区域克隆了12个基因,其中11个基因是基于Brassica rapaa数据库的注释,1个是功能未知的基因。拟南芥基因ATNYE1的一个同源基因Bra019346被确定为Brnye1的潜在候选基因。序列分析显示,在Bra019346innye的第二个外显子上插入了一个40bp的插入,该插入产生了TAA终止密码子。在1561个F2个体中,一个候选基因特异的Indel标记与突变体中的Brnye1表现出完美的共分离。这些结果为揭示小白菜保绿性状的分子机制奠定了基础。
Key messageUsing bulked segregant analysis combined with next-generation sequencing, we delimited theBrnye1gene responsible for the stay-green trait ofnyein pakchoi. Sequence analysis identifiedBra019346as the candidate gene.Abstract“Stay-green” refers to a plant trait whereby leaves remain green during senescence. This trait is useful in the cultivation of pakchoi (Brassica campestrisL. ssp.chinensis), which is marketed as a green leaf product. This study aimed to identify the gene responsible for the stay-green trait in pakchoi. We identified a stay-green mutant in pakchoi, which we termed “nye”. Genetic analysis revealed that the stay-green trait is controlled by a single recessive gene,Brnye1. Using the BSA-seq method, a 3.0-Mb candidate region was mapped on chromosome A03, which helped us localizeBrnye1to an 81.01-kb interval between SSR markers SSRWN27 and SSRWN30 via linkage analysis in an F2population. We identified 12 genes in this region, 11 of which were annotated based on theBrassica rapaannotation database, and one was a functionally unknown gene. An orthologous gene of theArabidopsisgeneAtNYE1,Bra019346, was identified as the potential candidate forBrnye1. Sequence analysis revealed a 40-bp insertion in the second exon ofBra019346innye, which generated the TAA stop codon. A candidate gene-specific Indel marker in 1561 F2individuals showed perfect cosegregation withBrnye1in thenyemutant. These results provide a foundation for uncovering the molecular mechanism of the stay-green trait in pakchoi.