Recalcitrant nature is a major constraint for the in vitro regeneration and genetic transformation of leguminous species members. Therefore, an improved genetic transformation in horse gram has been developed via in planta method, in which Agrobacterium strain harboring binary vector pCAMBIA2301 was used for the transformation. Several factors affecting in planta transformations were put forth viz. Agrobacterium cell density, co-cultivation, and sonication combined with vacuum infiltration duration which were optimized. Germinated seeds were sonicated and vacuum infiltrated with different densities of Agrobacterium culture and co-cultivated in half-strength MS medium with 100 μM of acetosyringone for 48 h. Seedlings were washed with cefotaxime and sowed in vermiculite soil for maturation. T1 plants were subjected to histochemical and molecular analysis to ensure transformation efficiency. Among various combinations analyzed, maximum transformation efficiency (20.8%) was attained with seeds of 5 min sonication combined with vacuum infiltration with 0.6 optical density of Agrobacterium culture. It concludes that a different Agrobacterium cell density with sonication combined with vacuum infiltration has improved transgenic efficiency in horse gram plants. This simple and efficient method is feasible for the stable expression of foreign genes that could be beneficial for future food security.

中文翻译：

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一种简单有效的农杆菌介导的马克植物转化方案（Macromylyloma uniflorum Lam。Verdc。）

顽calc性是豆科物种成员体外再生和遗传转化的主要障碍。因此，已经通过植物方法开发了改进的以马为单位的遗传转化，其中将携带二元载体pCAMBIA2301的农杆菌菌株用于转化。提出了影响植物转化的几个因素。农杆菌细胞密度，共培养和超声处理与真空渗透持续时间相结合进行了优化。对发芽的种子进行超声处理，并用不同密度的土壤杆菌培养物进行真空浸润，并在含有100μM乙酰丁香酮的半强度MS培养基中共培养48 h。幼苗用头孢噻肟洗涤并播种在ver石土壤中以使其成熟。对T1植物进行组织化学和分子分析以确保转化效率。在分析的各种组合中，超声处理5分钟的种子与农杆菌培养物的0.6光学密度的真空浸渗结合获得了最高转化效率（20.8％）。结论是，通过超声处理与真空浸渗相结合的不同土壤杆菌细胞密度提高了马氏植物的转基因效率。这种简单有效的方法对于稳定表达外源基因是可行的，这可能对未来的粮食安全有利。6.农杆菌培养的光密度。结论是，通过超声处理与真空浸渗相结合的不同土壤杆菌细胞密度提高了马氏植物的转基因效率。这种简单有效的方法对于稳定表达外源基因是可行的，这可能对未来的粮食安全有利。6.农杆菌培养的光密度。结论是，通过超声处理与真空浸渗相结合的不同土壤杆菌细胞密度提高了马氏植物的转基因效率。这种简单有效的方法对于稳定表达外源基因是可行的，这可能对未来的粮食安全有利。