Rapid identification of the purple stem (Ps) gene of Chinese kale (Brassica oleracea var. alboglabra) in a segregation distortion population by bulked segregant analysis and RNA sequencing
Rapid identification of the purple stem (Ps) gene of Chinese kale (Brassica oleracea var. alboglabra) in a segregation distortion population by bulked segregant analysis and RNA sequencing
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通过批量分离分析和 RNA 测序快速鉴定分离扭曲群体中羽衣甘蓝 (Brassica oleracea var. alboglabra) 的紫色茎 (Ps) 基因
DOI：
10.1007/s11032-017-0752-3
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发表时间：
2017-12-01
影响因子：
3.1
通讯作者：
Yan, Chenghuan
中科院分区：
文献类型：
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作者：
Tang, Qiwei;Tian, Mengyu;Yan, Chenghuan
丰富的花青素积累在许多花、叶、果实和茎中。本研究以紫茎芥和绿茎芥杂交的F-2分离群体为材料，分析了紫茎芥和绿茎芥的茎色遗传基础。采用散装分离分析结合RNA测序（BSR-seq）对紫色茎（Ps）的致病基因进行了遗传定位，结果表明，两群等位基因的频率差异在C09染色体上呈单峰。用406 kb序列将Ps精细定位到0.32 cM区间。二氢黄酮醇4-还原酶（DFR）是花青素生物合成途径中的一种酶，其编码基因在两个基因池中存在差异表达，并被确定为芥蓝紫色茎的候选基因。序列分析表明，在甘蓝BoDRF基因外显子2的编码序列中插入了1 bp，导致移码和功能丧失。BoDFR基因的表达量显著低于紫茎芥。Ps基因的遗传结果将为紫色蔬菜的进一步开发提供参考。
Abundant anthocyanins accumulate in many flowers, leaves, fruits, as well as stems. In this study, an F-2 segregating population derived from a cross between Chinese kale with a purple stem and kale with a green stem was used to decipher the genetic basis for the stem color. Bulked segregant analysis in combination with RNA sequencing (BSR-seq) was used to genetically map the causal gene for the purple stem (Ps), and the allele frequency difference between the two pools showed a single peak on chromosome C09. Ps was fine-mapped to the 0.32 cM interval with 406 kb sequences. A gene-encoding dihydroflavonol 4-reductase (DFR), an enzyme in the anthocyanin biosynthesis pathway, was differentially expressed between the two pools, and was identified as the candidate gene for the purple stem in Chinese kale. Sequence analysis revealed that 1 bp was inserted in the coding sequence of exon 2 of the BoDRF gene in green kale, causing frameshift and loss of function. The expression level of the BoDFR gene was significantly lower than the Chinese kale cultivar with a purple stem. The genetic results of the Ps gene will be useful for future development of purple vegetables.