Efficient plant regeneration via direct organogenesis and Agrobacterium tumefaciens-mediated genetic transformation of Picrorhiza kurroa: an endangered medicinal herb of the alpine Himalayas

被引：16

作者：

Bhat, Wajid Waheed ^{[1
]}

Lattoo, Surrinder K. ^{[1
]}

Rana, Satiander ^{[1
]}

Razdan, Sumeer ^{[1
]}

Dhar, Niha ^{[1
]}

Dhar, Rekha S. ^{[1
]}

Vishwakarma, Ram A. ^{[1
]}

机构：

[1] CSIR, Indian Inst Integrat Med, Jammu 180001, India

来源：

IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-PLANT | 2012年 / 48卷 / 03期

关键词：

Acetosyringone; Agrobacterium tumefaciens; Genetic transformation; Organogenesis; Picrorhiza kurroa; Picrosides; ROYLE EX BENTH; CELLS; PROPAGATION;

D O I：

10.1007/s11627-012-9434-3

中图分类号：

Q94 [植物学];

学科分类号：

071001 ;

摘要：

Picrorhiza kurroa Royle ex. Benth. is a medicinal herb of immense therapeutic value with restricted geographic distribution. Efficient plant regeneration via direct organogenesis and Agrobacterium tumefaciens-mediated genetic transformation was developed for this plant. Multiple shoot bud induction was achieved from leaf explants cultured in Gamborg's B-5 medium containing 3 % (w/v) sucrose, 3 mg/l kinetin and 1 mg/l indole-3-butyric acid. More than 90 % of leaf explants formed shoot buds leading to whole plant regeneration. An Agrobacterium-mediated genetic transformation protocol was developed using A. tumefaciens strain GV3101 harboring binary vector pCAMBIA1302 containing the green fluorescent protein and hygromycin phosphotransferase genes. Leaf explants precultured for 2 d were the most suitable for co-cultivation with Agrobacterium and transformation efficiency was enhanced with 200 mu M acetosyringone. Putative transformants were selected using media containing 15 mg/l hygromycin. Transformation was verified by detection of the green fluorescent protein using fluorescence microscopy and by polymerase chain reaction. Approximately 56 % of the explants were transformed with an average of 3.4 +/- 0.4 transgenic plantlets per explant. An efficient regeneration and transformation protocol thus developed enabling a fresh perspective of metabolic engineering in P. kurroa using an Agrobacterium-mediated transformation. This is the first report of direct organogenesis from leaf explants and genetic transformation of P. kurroa.

引用

页码：295 / 303

页数：9

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