a Laboratory of Translational Research on Transgenic Crops, Department of Botany , University of Calcutta , Kolkata , India.
b Crop Improvement Division , ICAR-National Rice Research Institute , Cuttack , India.
GM Crops Food. 2019;10(2):115-138. doi: 10.1080/21645698.2019.1625653. Epub 2019 Jun 12.
An efficient genetic transformation system is a prerequisite for studying gene functions, molecular breeding program, and introducing new traits. mediated genetic transformation is a widely preferred and accepted method for many plants, including pigeon pea. However, the efficiency of transformation of pigeon pea using the existing protocols is low and time-consuming. In the present study, we developed a rapid and highly efficient transformation system of pigeon pea, using embryonic axis-attached cotyledons as explants. We systematically investigated the influence of varying optical densities of suspension, duration of incubation, and co-cultivation on the transformation efficiency. In our system, a transformation efficiency of approximately 83% was achieved using cells at an optical density (OD) of 0.25, infection time of 15 min, and co-culturing with explants for 72 h in the light with 100µM acetosyringone. The entire procedure, starting from seed to establishment of transformed plants in soil, was achieved in 35-40 days. This is a rapid and highly efficient protocol for -mediated transformation of pigeon pea, which could potentially be a useful reference, not only for the genetic improvement of pigeon pea but also for other recalcitrant leguminous plants.
一个高效的遗传转化系统是研究基因功能、分子育种计划和引入新特性的前提。介导的遗传转化是许多植物(包括兵豆)广泛采用和接受的方法。然而,利用现有的方案进行兵豆的转化效率低且耗时。在本研究中,我们使用胚轴附着子叶作为外植体,开发了一种快速高效的兵豆遗传转化系统。我们系统地研究了悬浮液的不同光密度、孵育时间和共培养对转化效率的影响。在我们的系统中,使用光密度为 0.25、感染时间为 15 分钟的细胞,在有 100µM 乙酰丁香酮的光下共培养 72 小时,转化效率约为 83%。从种子到在土壤中建立转化植物的整个过程在 35-40 天内完成。这是一种快速高效的兵豆介导遗传转化方法,不仅对兵豆的遗传改良有潜在的参考价值,对其他难处理的豆科植物也有参考价值。