Major biotic and abiotic stresses have led to the reduction of chickpea productivity, creating a strong barrier for its utilization as major food legume. Genetic transformation has revolutionized the chickpea improvement programs for biotic stress tolerance. Recent knowledge on genomic resources offers a range of approaches for genetic improvement of chickpea with a greater resolution, but relies on the transgenic establishment which is the major bottleneck in this process. Poor tissue culture responses along with insensitivity to in vitro rooting hindered the transgenic chickpea development through tissue culture based transformation methods. In the present report, Agrobacterium-mediated plumular meristem transformation was adapted in chickpea to bypass the recalcitrancy of regenerable tissue. Embryonic axis of immature seedlings of chickpea cultivar was decapitated followed by pricking of plumular meristem present in shoot apex and cotyledonary nodes. Pricking is followed by infection by Agrobacterium tumefaciens carrying binary vector pBI121. Three days co-cultivation was performed with infected explants in 6-benzylaminopurine and α-naphthaleneacetic acid supplemented modified Murashige and Skoog medium. Transient GUS expression was observed in co-cultivated explants and PCR based screening strategy allowed the establishment of primary transgenic events with a frequency of 60%. Kanamycin mediated stringent selection of T₁ events helped to eliminate the chimeric plants and 44% of T₁ progenies were confirmed through PCR. Further, Southern hybridization was performed to identify transgene integration in T₁ events. Additionally, prominent GUS activity in T₁ events confirmed the expression of transgene. Plumular transformation method, based on culture dependent Agrobacterium-infection and culture independent plant selection as well as establishment, was reported for the first time in chickpea. This transformation method will boost the recovery of novel genotypes with improved agronomic traits and mutant development for functional genomics in chickpea.

主要的生物和非生物胁迫导致鹰嘴豆的生产力下降，为将其用作主要食品豆科植物创造了强大的障碍。遗传转化彻底改变了鹰嘴豆改善程序的生物胁迫耐受性。关于基因组资源的最新知识为鹰嘴豆的遗传改良提供了更高分辨率的一系列方法，但依赖于转基因建立，这是此过程中的主要瓶颈。通过基于组织培养的转化方法，不良的组织培养反应以及对体外生根的不敏感性阻碍了转基因鹰嘴豆的发育。在本报告中，农杆菌介导的羽状分生组织转化适用于鹰嘴豆，以绕过可再生组织的顽固性。鹰嘴豆品种未成熟幼苗的胚轴被断头，然后刺出茎尖和子叶节中的羽状分生组织。刺破后，携带双元载体pBI121的根癌农杆菌感染。与感染的外植体在6-苄基氨基嘌呤和α-萘乙酸补充的改良Murashige和Skoog培养基中进行三天共培养。在共培养的外植体中观察到瞬时GUS表达，并且基于PCR的筛选策略允许以60％的频率建立初级转基因事件。卡那霉素介导的T _1的严格选择这些事件有助于消除嵌合植物，并通过PCR证实了44％的T ₁后代。此外，进行Southern杂交以鉴定T ₁事件中的转基因整合。另外，在T ₁事件中突出的GUS活性证实了转基因的表达。首次报道了鹰嘴豆基于培养依赖农杆菌感染和不依赖培养的植物选择以及建立的整体转化方法。这种转化方法将促进具有改善的农艺性状和鹰嘴豆功能基因组学突变体开发的新基因型的恢复。