红麻过氧化物酶基因分离、过表达载体构建及转化拟南芥分析

发布时间：2019-04-17 09:23

【摘要】：红麻是一种以收获韧皮纤维为主的经济作物,因其具有纤维产量高,抗逆境生长能力强等特点,在我国大部分地区都有种植。近年来,随着其韧皮纤维在纺织工业中的应用,红麻越来越受到育种家的高度重视。其韧皮纤维中木质素含量高低是限制其纤维品质好坏的重要因素之一,过氧化物酶基因在不同品质的木材中差异表达较为显著。为研究该基因在红麻(Hibiscus cannabinus L.)木质素合成过程中的作用,通过红麻转录组数据库筛选到过氧化物酶(Peroxidase)基因的核心片段,利用Tail-PCR技术获得了Peroxidase基因c DNA全长,命名为Hc POD。生物信息学分析表明:Hc POD基因编码序列长952 bp,编码316个氨基酸,相对分子量为33.9 k D,等电点为4.85。为进一步研究该基因的功能,本研究利用p CAMBIA1301载体作为骨架载体构建了红麻过氧化物酶基因过表达载体,利用花絮侵染法转化拟南芥。通过对转基因和非转基因植株体内木质素含量检测后发现,在拟南芥体内过表达红麻POD基因能显著提高拟南芥的木质素含量水平。研究结果为进一步分析该基因功能和了解红麻木质素合成的分子机制具有意义。
[Abstract]:Kenaf is a kind of cash crop which mainly harvest phloem fiber. It has been planted in most areas of our country because of its characteristics of high fiber yield and strong growth resistance to adversity. In recent years, with the application of bast fiber in textile industry, kenaf has been paid more and more attention by breeders. The lignin content in the phloem fiber is one of the important factors limiting the quality of the fiber, and the expression of peroxidase gene in wood with different quality is significant. In order to study the gene in kenaf (Hibiscus cannabinus L.) In the process of lignin synthesis, the core fragment of peroxidase (Peroxidase) gene was screened by kenaf transcript database. The full length of Peroxidase gene c DNA, named Hc POD., was obtained by Tail-PCR technique. Bioinformatics analysis showed that the coding sequence of: Hc POD gene was 952 bp, encoding 316 amino acids with a relative molecular weight of 33.9 KD and an isoelectric point of 4.85. In order to further study the function of the gene, the p-CAMBIA1301 vector was used as the skeleton vector to construct the over-expression vector of kenaf peroxidase gene, and the transgenic Arabidopsis thaliana was transformed into Arabidopsis thaliana by the method of floss infection. By detecting the lignin content in transgenic and non-transgenic plants, it was found that the overexpression of kenaf POD gene in Arabidopsis thaliana significantly increased the lignin content in Arabidopsis thaliana. The results are of significance for further analysis of the function of the gene and understanding of the molecular mechanism of lignin synthesis in kenaf.
【作者单位】：上饶师范学院生命科学学院;中国农业科学院麻类研究所;
【基金】：江西省自然科学基金(20142BAB204025) 江西省教育厅科学技术研究项目(GJJ151050) 农业部麻类生物学与加工重点实验室2016年度开放课题(201602) 上饶师范学院2014年博士科研启动基金(001055)共同资助
【分类号】：Q943.2;S563.5

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