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Development of a rapid and highly efficient Agrobacterium-mediated transformation system for pigeon pea [Cajanus cajan (L.) Millsp].
GM Crops & Food ( IF 4.5 ) Pub Date : 2019-06-12 , DOI: 10.1080/21645698.2019.1625653
Subhasis Karmakar 1 , Kutubuddin Ali Molla 2, 3 , Dipak Gayen 4 , Aritra Karmakar 1 , Kaushik Das 1 , Sailendra Nath Sarkar 1 , Karabi Datta 1 , Swapan K Datta 1
Affiliation  

An efficient genetic transformation system is a prerequisite for studying gene functions, molecular breeding program, and introducing new traits. Agrobacterium tumefaciens-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 Agrobacterium suspension, duration of incubation, and co-cultivation on the transformation efficiency. In our system, a transformation efficiency of approximately 83% was achieved using Agrobacterium cells at an optical density (OD600) 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 Agrobacterium-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.

中文翻译:

快速高效农杆菌介导的木豆转化系统的开发[Cajanus cajan(L.)Millsp]。

一个有效的遗传转化系统是研究基因功能,分子育种程序和引入新特性的先决条件。根癌农杆菌介导的遗传转化是许多植物(包括木豆)的广泛优选和公认的方法。然而,使用现有协议对木豆进行转化的效率低且耗时。在当前的研究中,我们开发了一种快速高效的木豆转化系统,使用带有胚轴的子叶作为外植体。我们系统地研究了土壤杆菌悬液的不同光密度,孵育时间和共培养对转化效率的影响。在我们的系统中 使用土壤杆菌,光密度(OD600)为0.25,感染时间为15分钟,与外植体在100µM乙酰丁香酮下共培养72小时,转化效率约为83%。从种子到在土壤中转化植物的建立,整个过程在35至40天内完成。这是一种快速高效的农杆菌介导的木豆转化方法,不仅对木豆的遗传改良,而且对其他顽固的豆科植物都有潜在的参考价值。
更新日期:2019-11-01
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