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A highly efficient callus model to study gene functions: HbSRPP1 may play a role in the elongation of cis-1,4-polyisoprene in the rubber tree

文献类型： 外文期刊

作者： Tan, Deguan 1 ; Guo, Zihan 1 ; Fu, Lili 1 ; Yu, Ying 1 ; Peng, Jing 1 ; Huang, Yuchun 1 ; Zhou, Xue 1 ; Sun, Xuepiao 1 ; Zhang, Jiaming 1 ;

作者机构： 1.Chinese Acad Trop Agr Sci, Inst Trop Biosci & Biotechnol, Natl Key Lab Trop Crop Breeding, Key Lab Biol & Genet Resources Trop Crops,Sanya Re, Haikou, Peoples R China

2.Hainan Univ, Sch Trop Crops, Haikou 570228, Hannan, Peoples R China

3.Huazhong Agr Univ, Coll Plant Sci & Technol, Wuhan 430070, Peoples R China

关键词： Rubber tree; Callus model; Laticifer cell; Rubber biosynthesis; Function study; Laticifer-specific promoter; HbSRPP1

期刊名称：INDUSTRIAL CROPS AND PRODUCTS （ 影响因子：6.2； 五年影响因子：6.2 ）

ISSN： 0926-6690

年卷期： 2025 年 225 卷

页码：

收录情况： SCI

摘要： Natural rubber (NR) is synthesized in the laticifer cells of the rubber tree by a series of enzymes located on the membrane of the rubber particles (RPs). Traditional methods for function studies of NR biosynthesis-related genes are of low efficiency due to difficulties in obtaining transgenic plants and years of research cycle. We established a callus model for function studies without the need to regenerate transgenic plants. The callus of the rubber tree differentiated into laticifer cells efficiently with the induction by jasmonic acid, and RPs were observed by histochemical analysis. The rubber isolated from the callus was cis-1,4-polyisoprene with molecular weights similar to the rubber tapped from the tree. Laticifer-specific expression vectors of GUS and EGFP was constructed, and target genes can be easily inserted between the promoter and the reporter genes by Nimble cloning. HbSRPP1 gene was used as an example to test the efficiency of the callus model. HbSRPP1 cDNA was cloned in the laticifer-specific and EGFP-tagged expression vector, and transformed into the callus. The expression of HbSRPP1 gene was significantly up-regulated in the transgenic callus, and EGFP-tagged HbSRPP1 protein was successfully produced and dispatched onto the RPs. As a consequence, the content and molecular weight of the rubber were increased significantly in the transgenic callus, indicating that HbSRPP1 may play a role in the elongation of the cis-1,4-polyisoprene chain of the rubber and thus increase the rubber content. These results indicate that the callus can serve as an effective model for studying biological functions of genes that are related to laticifer development and rubber biosynthesis without the need to regenerate transgenic plants, and thus may accelerate functional genomics research and molecular breeding in the rubber tree.

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