Kola Vijaya Sudhakara Rao, Renuka P, Padmakumari Ayyagari Phani, Mangrauthia Satendra K, Balachandran Sena M, Ravindra Babu V, Madhav Maganti S
Department of Biotechnology, Indian Council of Agricultural Research-Indian Institute of Rice Research Hyderabad, India.
Department of Entomology, Indian Council of Agricultural Research-Indian Institute of Rice Research Hyderabad, India.
Front Physiol. 2016 Feb 12;7:20. doi: 10.3389/fphys.2016.00020. eCollection 2016.
RNAi is a powerful tool to target the insect genes involved in host-pest interactions. Key insect genes are the choice for silencing to achieve pest derived resistance where resistance genes are not available in gene pool of host plant. In this study, an attempt was made to determine the effect of dsRNA designed from two genes Cytochrome P450 derivative (CYP6) and Aminopeptidase N (APN) of rice yellow stem borer (YSB) on growth and development of insect. The bioassays involved injection of chemically synthesized 5' FAM labeled 21-nt dsRNA into rice cut stems and allowing the larvae to feed on these stems which resulted in increased mortality and observed growth and development changes in larval length and weight compared with its untreated control at 12-15 days after treatment. These results were further supported by observing the reduction in transcripts expression of these genes in treated larvae. Fluorescence detection in treated larvae also proved that dsRNA was readily taken by larvae when fed on dsRNA treated stems. These results from the present study clearly show that YSB larvae fed on dsRNA designed from Cytochrome P450 and Aminopeptidase N has detrimental effect on larval growth and development. These genes can be deployed to develop YSB resistance in rice using RNAi approach.
RNA干扰是一种靶向参与宿主与害虫相互作用的昆虫基因的强大工具。关键昆虫基因是进行沉默的选择对象,以便在宿主植物基因库中不存在抗性基因的情况下实现害虫衍生抗性。在本研究中,尝试确定从水稻二化螟(YSB)的两个基因细胞色素P450衍生物(CYP6)和氨肽酶N(APN)设计的dsRNA对昆虫生长发育的影响。生物测定包括将化学合成的5'FAM标记的21核苷酸dsRNA注射到水稻切段茎中,并让幼虫取食这些茎,结果导致死亡率增加,并且与处理后12 - 15天的未处理对照相比,观察到幼虫长度和重量的生长发育变化。通过观察处理后幼虫中这些基因转录本表达的降低,进一步支持了这些结果。在处理后的幼虫中的荧光检测也证明,当幼虫取食dsRNA处理的茎时,dsRNA很容易被幼虫摄取。本研究的这些结果清楚地表明,取食从细胞色素P450和氨肽酶N设计的dsRNA的水稻二化螟幼虫对幼虫生长发育有有害影响。这些基因可用于利用RNA干扰方法培育水稻对二化螟的抗性。