数字农科院2.0

Identification, charectrization and genetic transformation of lignin and pectin polysaccharides through CRISPR/Cas9 in Nicotiana tobacum

文献类型： 外文期刊

作者： Rana Imtiaz Ahmed;Angyan Ren;Dalal Sulaiman Alshaya;Sajid Fiaz;Yingzhen Kong;Shoaib Liaqat;Naushad Ali;Muhammad Abu Bakar Saddique;Kotb A. Attia;Mahmood Ul Hassan Taga

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关键词： Agrobacterium-mediated transformation;Biopolymers;Homogalacturonan;Lignin;Pectin;Polysaccharides;Primary cell wall

期刊名称： Functional and Integrative Genomics

ISSN： 1438-793X

年卷期： 2024 年 24 卷 5 期

页码：

收录情况： SCIE（2024版）

摘要： CRISPR/Cas9 system has been successfully implemented in animals and plants is a second-generation genome editing tool. We are able to optimize a Cas9 system to edited Ntab06050 and Ntab0857410 genes in HD and K326 tobacco cultivars respectively. The gene Ntab06050 is related to lignin synthesis while the gene Ntab0857410 belongs to pectin synthesis by utilizing Agrobacterium-mediated leaf disc method. We have constructed total eight different constructs for the lignin related gene family CCoAMT, out of which three constructs have been selected from Ntab0184090, two constructs from Ntab0392460 while one construct from each Ntab0540120, Ntab0857410 and Ntab0135940 gene. To study the Cas9 system in pectin related genes, total five constructs have been utilized under Cas9 system and multiple target sites were selected by identifying PAM sequences. Out of which three constructs were targeted from NtabGAE1and NtabGAE6 homologous while two were targeted from NtabGAUT4 homologous. Where as, UDP-D-glucuronate 4-epimerase gene family is a Golgi localized, might have a role in the interconvertion of UDP-D-GlcA and UDP-D-GalA in pectin synthesis. We have succeeded in the mutation of pectin related NtabGAUT4 and lignin related NtabCCoAMT genes with 6.2% and 9.4% mutation frequency.

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