Banerjee Sagar, Gill Sarvajeet S, Gawade Bharat H, Jain Pradeep K, Subramaniam Kuppuswamy, Sirohi Anil
Division of Nematology, ICAR-Indian Agricultural Research Institute, New Delhi, India.
Stress Physiology and Molecular Biology Lab, Centre for Biotechnology, Maharshi Dayanand University, Rohtak, India.
Front Plant Sci. 2018 Jan 22;8:2266. doi: 10.3389/fpls.2017.02266. eCollection 2017.
Root-knot nematodes have emerged as devastating parasites causing substantial losses to agricultural economy worldwide. Tomato is the most favored host for major species of root-knot nematodes. Control strategies like use of nematicides have proved to be harmful to the environment. Other control methods like development of resistant cultivars and crop rotation have serious limitations. This study deals with the application of host generated RNA interference toward development of resistance against root-knot nematode in tomato. Two cuticle collagen genes viz. and involved in the synthesis and maintenance of the cuticle in were targeted through host generated RNA interference. Expression of both and was found to be higher in adult females followed by egg masses and J2s. Tomato var. Pusa Ruby was transformed with the RNAi constructs of these genes to develop transgenic lines expressing the target dsRNAs. 30.80-35.00% reduction in the number of adult females, 50.06-65.73% reduction in the number of egg mass per plant and 76.47-82.59% reduction in the number of eggs per egg mass were observed for the T events expressing dsRNA. Similarly, 34.14-38.54% reduction in the number of adult females, 62.34-66.71% reduction in number of egg mass per plant and 67.13-79.76% reduction in the number of eggs per egg mass were observed for the T generation expressing dsRNA. The multiplication factor of reduced significantly in both the cases and the structure of adult females isolated from transgenic plants were heavily distorted. This study demonstrates the role of the cuticle collagen genes and in the structure and development of cuticle inside the host and reinforces the potential of host generated RNA interference for management of plant parasitic nematodes (PPNs).
根结线虫已成为极具破坏力的寄生虫,给全球农业经济造成了巨大损失。番茄是主要根结线虫种类最喜爱的寄主。事实证明,使用杀线虫剂等防治策略对环境有害。其他防治方法,如培育抗性品种和轮作,也存在严重局限性。本研究探讨了利用寄主产生的RNA干扰来培育番茄抗根结线虫的抗性。通过寄主产生的RNA干扰,靶向了两个参与根结线虫表皮合成和维持的表皮胶原蛋白基因,即 和 。发现 和 在成年雌虫中的表达较高,其次是卵块和J2期幼虫。用这些基因的RNAi构建体转化番茄品种Pusa Ruby,以培育表达目标dsRNA的转基因株系。对于表达 dsRNA的T代植株,观察到成年雌虫数量减少30.80 - 35.00%,单株卵块数量减少50.06 - 65.73%,每个卵块中的卵数量减少76.47 - 82.59%。同样,对于表达 dsRNA的T代植株,观察到成年雌虫数量减少34.14 - 38.54%,单株卵块数量减少62.34 - 66.71%,每个卵块中的卵数量减少67.13 - 79.76%。在这两种情况下,根结线虫的繁殖系数均显著降低,从转基因植物中分离出的成年雌虫结构严重变形。本研究证明了表皮胶原蛋白基因 和 在寄主内根结线虫表皮结构和发育中的作用,并强化了寄主产生的RNA干扰在植物寄生线虫(PPNs)防治中的潜力。