Identification and characterization of pectin related gene NbGAE6 through virus-induced gene silencing in Nicotiana benthamiana

文献类型： 外文期刊

第一作者： Ahmed, Rana Imtiaz

作者： Ahmed, Rana Imtiaz;Ren, Angyan;Ding, Anming;Yang, Dahai;Kong, Yingzhen;Ahmed, Rana Imtiaz

作者机构：

关键词： GAE family; Homogalacturunan; N. benthamiana; Primary cell wall; VIGS

期刊名称：GENE （ 影响因子：3.688； 五年影响因子：3.329 ）

ISSN： 0378-1119

年卷期： 2020 年 741 卷

页码：

收录情况： SCI

摘要： Virus-induced gene silencing (VIGS) is a transient based reverse genetic tool used to elucidate the function of novel gene in N. benthamiana. In current study, 14 UDP-D-glucuronate 4-epimerase (GAE) family members were identified and their gene structure, phylogeny and expression pattern were analyzed. VIGS system was optimized for the functional characterization of NbGAE6 homologous genes in N. benthamiana. Whilst the GAE family is well-known for the interconversion of UDP-D-GlcA and UDP-D-GalA during pectin synthesis. Our results revealed that the downregulation of these genes significantly reduced the amount of GalA in the homogalacturunan which is the major component of pectin found in primary cell wall. Biphenyl assay and high performance liquid chromatography analysis (HPLC) depicted that the level of 'GalA' monosaccharide reduced to 40-51% in VIGS plants as compared to the wild type plants. Moreover, qRT-PCR also confirmed the downregulation of the NbGAE6 mRNA in VIGS plants. In all, this is the first comprehensive study of the optimization of VIGS system for the provision of rapid silencing of GAE family members in N. benthamiana, eliminating the need of stable transformants.

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