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Host-induced silencing of a nematode protease gene in tomato plants conferred enhanced resistance to root-knot nematodes

P root-knot nematodes (Meloidogyne spp.) are arguably the most damaging genus of biotrophic pests of vascular plants and thus have a major impact on global agricultural production. Due to the changing climate and agricultural practices RKNs are becoming a menace in newer crops and geographical localities. Currently available management practices have failed to contain the problem; hence, there is a critical need to develop environmentally-friendly and smart approaches tailor-made to reduce the nematode disease burden in Indian agriculture. Utility of host-delivered RNAi has been demonstrated in several plants (Arabidopsis, tobacco and soybean) that exhibited resistance against root-knot and cyst nematodes. In the present study, a M. incognita-specific protease gene, cathepsin L cysteine proteinase (Mi-cpl-1) was targeted to generate tomato transgenic lines to evaluate the genetically modified nematode resistance. In vitro knockdown of Mi-cpl-1 gene led to the reduced attraction and penetration of M. incognita in tomato suggesting the involvement of Mi-cpl-1 in nematode parasitism. Transgenic expression of the dsRNA of Mi-cpl-1 gene resulted in 60-80% reduction in infection and multiplication of M. incognita in tomato. Evidence for in vitro and in planta silencing of Mi-cpl-1 was confirmed by expression analysis using quantitative RT-PCR. Our study demonstrates that Mi-cpl-1 plays crucial role during plant-nematode interaction and plantmediated down regulation of this gene elicits detrimental effect on M. incognita development, reinforcing the potential of RNAi technology for management of phytonematodes in crop plants. The findings of the present study lead to the better understanding of the mechanism of nematode parasitism which ultimately helps in designing smarter nematode management options.