Kesiraju Karthik, Mishra Pragya, Bajpai Akansha, Sharma Manju, Rao Uma, Sreevathsa Rohini
ICAR-National Institute for Plant Biotechnology, Pusa Campus, New Delhi, 110012 India.
Amity Institute of Biotechnology, Amity University Haryana, Manesar, Gurugram, 122413 India.
Physiol Mol Biol Plants. 2020 Nov;26(11):2319-2327. doi: 10.1007/s12298-020-00887-y. Epub 2020 Oct 19.
Cotton ( L.), a mercantile crop plant, is grown worldwide for fiber and seed oil. As with other economically important crops, cotton is bogged down with many biotic and abiotic stress factors. Towards this, genetic engineering offers numerous protocols to engineer plants for better resilience. However, recalcitrance of cotton to plant tissue culture has been the major constraint for successful in vitro regeneration. Hence, alternate methods that evade tissue culture regeneration have been envisaged. Non tissue culture-based transformation strategies are in vogue due to amenability and ease in the generation of transgenic plants. In the present study, we demonstrate the utility of an transformation protocol and establishment of a stringent selection agent-based screening for the identification of transgenics. The genotype independent nature of the protocol was validated in cotton cv. Pusa 8-6 using Preliminary transformation efficiency of 28% was achieved with a screening efficiency of 20% in the presence of hygromycin. The proof of T-DNA integration by various molecular and expression analysis in T1 and T2 generations proved that this technique can be employed to generate transgenic cotton.
棉花(棉属)是一种商业作物,在全球范围内种植以获取纤维和籽油。与其他经济上重要的作物一样,棉花受到许多生物和非生物胁迫因素的困扰。为此,基因工程提供了许多方案来对植物进行工程改造以提高其抗逆性。然而,棉花对植物组织培养的难处理性一直是其成功进行体外再生的主要限制因素。因此,人们设想了规避组织培养再生的替代方法。基于非组织培养的转化策略因易于操作且便于生成转基因植物而流行起来。在本研究中,我们展示了一种转化方案的实用性,并建立了基于严格选择剂的筛选方法来鉴定转基因植株。该方案不依赖基因型的特性在棉花品种Pusa 8 - 6中得到了验证,在潮霉素存在的情况下,初步转化效率达到28%,筛选效率为20%。通过T1和T2代的各种分子和表达分析证明T - DNA整合,这证明该技术可用于生成转基因棉花。