甘蓝型油菜蜡质相关基因的克隆与表达分析
发布时间：2018-12-23 12:37
【摘要】：植物叶片中,表皮蜡质对植物的生长发育和适应外界环境起着重要作用。本研究针对拟南芥(Arabidopsis thaliana)已发表的蜡质相关基因信息,基于生物信息学的方法获得以甘蓝型油菜(Brassica napus)品种中双11号为参考的10个蜡质相关基因的44个拷贝序列。然后,以在抽薹期前后叶片蜡质差异明显的3对自然突变的甘蓝型油菜(L1050,P144和W481)为材料,利用同源克隆的方法获得这些拷贝成员在3对甘蓝型油菜中的核苷酸和蛋白序列,通过多重比对分析发现,甘蓝型油菜蜡质基因(wax inducer 2,WIN2)家族中的Bn WIN2C01在核苷酸与氨基酸水平上均存在稳定的突变位点。利用q RT-PCR分析基因Bn WIN2C01在甘蓝型油菜突变体材料抽薹期前后叶片中的表达情况,结果表明该基因在表皮蜡质合成前后的叶片中达到极显著差异水平(P0.01),说明Bn WIN2C01基因的特异性表达可能影响了叶片蜡质的合成。本研究将有助于阐明叶表皮蜡质形成过程的分子机理,为提高油菜抗逆性提供一定的参考依据。
[Abstract]:In plant leaves, epidermal wax plays an important role in the growth and development of plants and adaptation to the external environment. In this study, based on bioinformatics, we obtained 44 copy sequences of 10 waxy related genes of Arabidopsis thaliana (Arabidopsis thaliana) based on bioinformatics, which were referenced by Zhongshuang11, a Brassica napus (Brassica napus) variety. Then, three pairs of natural mutant Brassica napus (L1050P144 and W481), which had obvious difference in waxy quality of leaves before and after bolting, were used as materials. The nucleotide and protein sequences of these copy members in 3 pairs of Brassica napus were obtained by homologous cloning. The waxy gene (wax inducer 2 of Brassica napus was found by multiple alignment analysis. Bn WIN2C01 in the WIN2 family had stable mutation sites at nucleotide and amino acid levels. Q RT-PCR analysis was used to analyze the expression of Bn WIN2C01 gene in the leaves of Brassica napus mutant before and after bolting stage. The results showed that the gene reached a very significant difference level in the leaves before and after epidermal waxy synthesis (P0.01). These results suggest that the specific expression of Bn WIN2C01 gene may affect the waxy synthesis in leaves. This study will be helpful to elucidate the molecular mechanism of the formation of epidermis waxy and provide some references for improving the stress resistance of rape.
【作者单位】： 西南大学农学与生物科技学院/重庆市油菜工程技术研究中心;西南大学农业科学研究院;
【基金】：国家重点基础研究发展计划(973)项目(No.2015CB150201) 国家自然科学基金项目(No.31401412,No.U1302266和No.31571701) 国家科技支撑计划项目(No.2013BAD01B03-12) 中央高校基本科研业务费专项基金(No.XDJK2016B030) 西南地区强优势杂种优势利用项目(No.2016YFD0101300)
【分类号】：Q943.2;S565.4
本文编号：2389993
[Abstract]:In plant leaves, epidermal wax plays an important role in the growth and development of plants and adaptation to the external environment. In this study, based on bioinformatics, we obtained 44 copy sequences of 10 waxy related genes of Arabidopsis thaliana (Arabidopsis thaliana) based on bioinformatics, which were referenced by Zhongshuang11, a Brassica napus (Brassica napus) variety. Then, three pairs of natural mutant Brassica napus (L1050P144 and W481), which had obvious difference in waxy quality of leaves before and after bolting, were used as materials. The nucleotide and protein sequences of these copy members in 3 pairs of Brassica napus were obtained by homologous cloning. The waxy gene (wax inducer 2 of Brassica napus was found by multiple alignment analysis. Bn WIN2C01 in the WIN2 family had stable mutation sites at nucleotide and amino acid levels. Q RT-PCR analysis was used to analyze the expression of Bn WIN2C01 gene in the leaves of Brassica napus mutant before and after bolting stage. The results showed that the gene reached a very significant difference level in the leaves before and after epidermal waxy synthesis (P0.01). These results suggest that the specific expression of Bn WIN2C01 gene may affect the waxy synthesis in leaves. This study will be helpful to elucidate the molecular mechanism of the formation of epidermis waxy and provide some references for improving the stress resistance of rape.
【作者单位】： 西南大学农学与生物科技学院/重庆市油菜工程技术研究中心;西南大学农业科学研究院;
【基金】：国家重点基础研究发展计划(973)项目(No.2015CB150201) 国家自然科学基金项目(No.31401412,No.U1302266和No.31571701) 国家科技支撑计划项目(No.2013BAD01B03-12) 中央高校基本科研业务费专项基金(No.XDJK2016B030) 西南地区强优势杂种优势利用项目(No.2016YFD0101300)
【分类号】：Q943.2;S565.4
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