An Efficient Hairy Root Transformation Method for Common Bean based on Petiole Explants

Y.H. Liu, X.Q. Cai, K. Ning, P. Xu
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Abstract

Background: Common bean (Phaseolus vulgaris L.) is a major food legume with high nutritional value and economic importance globally. A remarkable attribute of this plant is its propensity to generate adventitious roots from petioles of detached leaves when moisture is appropriate. This distinctive feature presents promising prospects for harnessing petioles in the facilitation of transgenic hairy root production. Methods: We achieved the successful induction of transgenic hairy roots from common bean petioles through the utilization of Agrobacterium rhizogenes strain K599. Our experimentation encompassed a diverse array of common bean varieties and leaves of varying ages. Subsequently, we refined and optimized the procedure for hairy root induction. Result: Our investigations have revealed that the most conducive conditions for hairy root transformation with K599 are attained using 5-day-old leaves from the cultivar ‘Honghuaqingjia’, exhibiting a remarkable induction efficiency of 59%. The usefulness of this system was demonstrated through a subcellular localization analysis of the transcription factor PvTCP2 protein in combination with GFP (Green Fluorescent Protein).
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基于叶柄外植体的普通豆高效毛根转化方法
背景:蚕豆(Phaseolus vulgaris L.)是一种主要的食用豆科植物,具有很高的营养价值和全球重要的经济价值。这种植物的一个显著特点是,当水分适宜时,它能从脱落叶片的叶柄上产生不定根。这一显著特点为利用叶柄促进转基因毛细根的生产带来了广阔的前景。方法:我们利用根瘤农杆菌 K599 株成功地从蚕豆叶柄中诱导出了转基因毛细根。我们的实验涵盖了多种蚕豆品种和不同年龄的叶片。随后,我们改进并优化了毛根诱导程序。结果:我们的研究表明,使用 K599 进行毛细根转化的最有利条件是栽培品种 "红华清香 "的 5 天龄叶片,其诱导效率高达 59%。通过对转录因子 PvTCP2 蛋白与 GFP(绿色荧光蛋白)的亚细胞定位分析,证明了该系统的实用性。
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