Novel?iridoviral kinase induces mortality and?reduces performance  of?green peach aphids (Myzus persicae) in?transgenic Arabidopsis plants

Saranya?Ganapathy; Megha?N.?Parajulee; Michael?San?Francisco; Hong?Zhang; Shan?L.?Bilimoria

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자료유형: 학술저널

저자정보: Saranya?Ganapathy (Texas Tech University) Megha?N.?Parajulee (Texas Tech University) Michael?San?Francisco (Texas Tech University) Hong?Zhang (Texas Tech University) Shan?L.?Bilimoria (Texas Tech University)

저널정보: 한국식물생명공학회 Plant Biotechnology Reports Plant Biotechnology Reports 제15권 제1호

발행연도: 2021.1

수록면: 13 - 25 (13page)

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Insect pests are a serious threat to agricultural productivity. Genetically-modifed crops ofer a promising alternative, and Bacillus thuringiensis endotoxin genes have played a major role in this respect. However, to overcome insect tolerance issues and to broaden the target range, it is critical to identify alternative insecticidal toxins working through novel mechanisms. In our previous study, a kinase from Chilo iridescent virus (CIV) that has insecticidal activity was identifed and designated as Iridovirus Serine/Threonine Kinase (ISTK). A 35?kDa truncated form of ISTK, designated iridoptin, was obtained during expression and purifcation of ISTK in the yeast system. This yeast-expressed CIV toxin induced 50% mortality in cotton aphids and 100% mortality in green peach aphids (GPA). In this study, codon optimized ISTK gene and iridoptin fragment (designated oISTK and oIRI, respectively) were designed by altering the codon usage features that are seldom present in plant exons. Codon-optimized gene(s) cloned into plant expression vectors were used to stably transform Arabidopsis plants. PCR analysis of genomic DNA of transformed plants confrmed the presence of the DNA insert (oISTK/oIRI) in selected transgenic lines. Further screening was performed by selecting the PCR positive lines, which showed expression of respective toxins at the polypeptide level, using Western blot analyses. Codon-optimized gene constructs resulted in signifcant improvement in levels of expression of ISTK and iridoptin polypeptide and confrmed its stability in planta. The stable lines expressing either of the two toxin forms induced moderate to very high mortality in GPAs and signifcantly afected GPA development and fecundity.

#Transgenic insect-resistant plants?· Green peach aphids?· Chilo iridescent virus?· Serine/threonine kinase?· Plant-incorporated protectants

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