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未熟抽薹会对大白菜(Brassica rapa ssp.pekinensis)生产造成重大损失。为研究大白菜抽薹开花相关基因,本研究对添加结球甘蓝(B.oleracea var.capitata)4号染色体片段的大白菜耐抽薹和易抽薹易位系间存在差异的c DNA-扩增片段长度多态性(c DNA-amplified fragment length polymorphism,c DNAAFLP)条带进行回收测序,其差异条带的同源性比对和功能性分析结果表明,差异基因可分为3种:转录翻译类、信号传导类和膜运输类。其功能预测涉及细胞物质运输、信号转导、物质能量代谢、胁迫应答、基因表达调控、细胞周期等。对其中8条序列进行q RT-PCR检测结果表明,在春化处理过程中P65M58-11、P65M58-12和P75M47-9在极晚抽薹材料中表达量峰值极显著高于极早抽薹材料,P65M58-11和P65M58-12条带的表达量峰值出现在春化处理14 d时。而P75M47-9在极晚抽薹材料中表达量峰值出现在春化处理7 d,在极早抽薹材料中从春化处理开始就呈现出下调趋势;P77M58-9、P77M58-12、P63M49-8-1和P63M61-4-2在极晚抽薹材料中表达量峰值极显著低于极早抽薹材料,P63M49-8-1在极早抽薹材料中表达量的峰值出现在春化处理7 d,而在极晚抽薹材料中从春化处理7 d后表达量逐渐上调。P77M58-9、P77M58-12和P63M61-4-2表达量峰值出现在春化处理14 d时;另外P63M49-1-2条带在极晚抽薹和极早抽薹两种材料中表达量差异无明显规律,在极早抽薹材料中表达量峰值出现在春化处理7 d时,而在极晚抽薹材料中峰值出现在春化处理21 d时。本研究获得了在6个易位系、2个自交系、2个商品种的耐抽薹和易抽薹材料间表达量及表达模式存在差异的8个基因,为深入挖掘大白菜抽薹开花相关基因、开展基因功能验证以及获得耐抽薹大白菜材料提供基础资料。
Unpolished bolting can cause significant losses in the production of Brassica rapa ssp. Pekinensis. In order to study the bolting and flowering related genes of Chinese cabbage, we studied the differences of the length of c DNA-amplified fragments between Chinese cabbage and boll-tolerant translocation lines with B.oleracea var.capitata chromosome 4 The sequences of the c DNA-amplified fragment length polymorphism (cDNAAFLP) were sequenced. The homology comparison and functional analysis of the differentially expressed bands showed that there are three types of differential genes: transcriptional translation, Signal transduction and membrane transport. Its functional prediction involves cellular material transport, signal transduction, material energy metabolism, stress response, gene expression regulation, cell cycle and so on. The results of q RT-PCR showed that the peak of expression level of P65M58-11, P65M58-12 and P75M47-9 in very late bolting material was significantly higher than that of early bolting material, and P65M58 The expression peak of -11 and P65M58-12 appeared in the vernalization 14 d. However, the peak of expression of P75M47-9 in the late bolting material appeared in the vernalization treatment for 7 days, and down-regulated in the early bolting material from the vernalization treatment. P77M58-9, P77M58-12, P63M49-8- 1 and P63M61-4-2 in extremely late bolting material were significantly lower than those in early bolting material. The peak of expression of P63M49-8-1 in early bolting material appeared in vernalization treatment for 7 days, whereas in The late bolting material gradually increased from 7 days after vernalization. P77M58-9, P77M58-12 and P63M61-4-2 peaked at vernalization 14 d. In addition, there was no significant difference in the expression of P63M49-1-2 between late bolting and early bolting The peak value of expression in the early bolting material appeared in the vernalization treatment for 7 d, while in the late bolting material, the peak appeared in the vernalization treatment for 21 d. In this study, we obtained eight genes that were different in the expression and expression pattern of bolting and bolting in six translocation lines, two inbred lines and two commercial varieties. In order to further explore the bolting and flowering genes of Chinese cabbage , To carry out gene function verification and access to bolting Chinese cabbage materials to provide basic information.