ICAR-National Institute for Plant Biotechnology, New Delhi, 110012, India.
ICAR-IARI, Division of Plant Physiology, New Delhi, 110012, India.
Planta. 2022 Sep 9;256(4):74. doi: 10.1007/s00425-022-03977-1.
Mi-msp10 and Mi-msp23 effector genes play a significant role during Meloidogyne incognita parasitism on Arabidopsis roots. The role of these genes was confirmed by demonstrating the decrease of the level of susceptibility of Arabidopsis by the silencing of Mi-msp10 and Mi-msp23 genes using HD-RNAi technology. Root-knot nematodes (RKNs) are the most damaging pathogens severely affecting global food production. The sustainable options to minimize menace of nematode populations through economically feasible measures are limited. Thus, the development of innovative and target-specific strategies that aid in their management is imperative. RNAi technology has emerged as a sustainable and target-specific alternative to control phytonematodes. Here, we characterized two novel subventral gland and dorsal gland-specific effectors, Mi-msp10 and Mi-msp23, to determine their potential effectiveness in controlling M. incognita. Comparative developmental profiling using qRT-PCR revealed higher expression of both effectors in the adult nematode female. Furthermore, functional evaluation of Mi-msp10 and Mi-msp23 dsRNA cassettes was performed using host-delivered RNAi (HD-RNAi) in Arabidopsis. The transgenic lines were examined against M. incognita, and the phenotypic effect of HD-RNAi was evident with a 61% and 51% reduction in gall formation in the Mi-msp10 and Mi-msp23 RNAi lines, respectively. A significant drop in the nematode adult females by 59% for Mi-msp10 and 49% for Mi-msp23-RNAi lines was observed. Similarly, production in egg masses decreased significantly by 76% (Mi-msp10) and 60% (Mi-msp23) for the RNAi lines, which eventually decreased the reproductive factor by 92% and 75%, respectively. The gene expression analysis showed a significant decrease in the transcript level by up to 72% (Mi-msp10) and 66% (Mi-msp23) in M. incognita females feeding on RNAi lines, providing further evidence of effective gene silencing. Overall, our findings provide useful information and support further development of RNAi-based strategies to control M. incognita.
Mi-msp10 和 Mi-msp23 效应基因在南方根结线虫寄生拟南芥根过程中发挥重要作用。通过使用 HD-RNAi 技术沉默 Mi-msp10 和 Mi-msp23 基因来降低拟南芥的易感性，证实了这些基因的作用。根结线虫（RKNs）是最具破坏性的病原体，严重影响全球粮食生产。通过经济可行的措施来最大限度地减少线虫种群威胁的可持续选择是有限的。因此，开发创新和靶向的策略来帮助管理它们是至关重要的。RNAi 技术已成为控制植物寄生线虫的一种可持续和靶向的替代方法。在这里，我们鉴定了两个新的亚腹腺和背腺特异性效应子 Mi-msp10 和 Mi-msp23，以确定它们在控制南方根结线虫中的潜在有效性。使用 qRT-PCR 进行的比较发育分析显示，这两个效应子在成年雌性线虫中表达更高。此外，使用拟南芥中的宿主传递 RNAi（HD-RNAi）对 Mi-msp10 和 Mi-msp23 dsRNA 盒进行了功能评估。对转基因株系进行了南方根结线虫的检测，HD-RNAi 的表型效应明显，Mi-msp10 和 Mi-msp23 RNAi 株系的根结形成分别减少了 61%和 51%。Mi-msp10-RNAi 株系的线虫成年雌性数量减少了 59%，Mi-msp23-RNAi 株系的线虫成年雌性数量减少了 49%。同样，Mi-msp10-RNAi 株系的卵块产量显著减少了 76%，Mi-msp23-RNAi 株系的卵块产量减少了 60%，最终使繁殖因子分别减少了 92%和 75%。基因表达分析显示，在取食 RNAi 株系的线虫雌性中，Mi-msp10 的转录水平显著下降了 72%，Mi-msp23 的转录水平下降了 66%，进一步证明了有效的基因沉默。总的来说，我们的研究结果提供了有用的信息，并支持进一步开发基于 RNAi 的策略来控制南方根结线虫。
