Abstract

AKT-interacting protein (AKTIP) interacts with serine/threonine protein kinase B (PKB)/AKT. AKTIP modulates AKT’s activity by enhancing the phosphorylation of the regulatory site and plays a crucial role in multiple biological processes. In this study, the full length cDNA of NlAKTIP, a novel AKTIP gene in the brown planthopper (BPH) Nilaparvata lugens, was cloned. The reverse transcription quantitive PCR (RT-qPCR) results showed that the NlAKTIP gene was strongly expressed in gravid female adults, but was relatively weakly expressed in nymphs and male adult BPH. In female BPH, treatment with dsAKTIP resulted in the efficient silencing of NlAKTIP, leading to a significant reduction of mRNA levels, about 50% of those of the untreated control group at day 7 of the study. BPH fed with dsAKTIP had reduced growth with lower body weights and smaller sizes, and the body weight of BPH treated with dsAKTIP at day 7 decreased to about 30% of that of the untreated control. Treatment of dsAKTIP significantly delayed the eclosion for over 7 days relative to the control group and restricted ovarian development to Grade I (transparent stage), whereas the controls developed to Grade IV (matured stage). These results indicated that NlAKTIP is crucial to the growth and development of female BPH. This study provided a valuable clue of a potential target NlAKTIP for inhibiting the BPH, and also provided a new point of view on the interaction between BPH and resistant rice.

Highlights

  • The brown planthopper (BPH) Nilaparvata lugens is a major insect pest for rice crops

  • Full-length NlAKTIP cDNA (GenBank Acc no: KP027545, not released) was cloned using the rapid amplification of cDNA ends (RACE) method according to our previous transcriptome sequence information

  • We found that NlAKTIP was down-regulated when the BPH was reared on resistant rice variety RHT, including different colonies of RF (RF1 to RF5) and Rh colony, suggesting that NlAKTIP

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Summary

Introduction

The brown planthopper (BPH) Nilaparvata lugens is a major insect pest for rice crops. Sustainable control of BPH is a serious problem, and to solve it requires exploring the interaction between BPH and resistant rice varieties, which includes revealing the molecular mechanism of BPH growth and development. We analyzed BPH gene expression profile [23] and found that a gene encoding AKT interacting protein (NlAKTIP) in our Rh colony [reared on resistant Rathu Heenati (RHT, with Bph 3). NlAKTIP might be an important responsive gene in BPH that feed on resistant rice. We intended to explore the function of NlAKTIP in BPH and expected our study to provide new clues for screening important genes as targets for pest control

Results
Effects of NlAKTIP Knockdown on mRNA and Protein Expression
Effects of NlAKTIP RNAi on Body Weight and Size
Effects of NlAKTIP RNAi on Eclosion and Ovarian Development
Effects of NlAKTIP RNAi on BPH Survival Rates
Discussion
Insects
Cloning of Full-Length NlAKTIP cDNA
Gene Expression Analysis of NlAKTIP
Synthesis of dsRNA
Insect Bioassays
Western Blot Analysis
Data Analysis
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