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1. Assessment of suitable reference genes for qRT-PCR analysis in Adelphocoris suturalis
LUO Jing, MA Chao, LI Zhe, ZHU Bang-qin, ZHANG Jiang, LEI Chao-liang, JIN Shuang-xia, J. Joe Hull, CHEN Li-zhen
Journal of Integrative Agriculture    2018, 17 (12): 2745-2757.   DOI: 10.1016/S2095-3119(18)61926-4
摘要258)      PDF(pc) (1312KB)(318)    收藏
Quantitative reverse transcription polymerase chain reaction (qRT-PCR) is the most commonly-used tool for measurement of gene expression, but its accuracy and reliability depend on appropriate data normalization with the use of one or more stable reference genes.  Adelphocoris suturalis is one of the most destructive pests of cotton, but until recently knowledge of its underlying molecular physiology had been hindered by a lack of molecular resources.  To facilitate research on this pest, we evaluated 12 common housekeeping genes studied in insects (GAPDH, ACT, βACT, TBP, SDH, βTUB, EF1γ, EF1α, EF1δ, RPL32, RPS15, and RPL27) for their expression stability in A. suturalis when subjected to various experimental treatments, including three biotic (developmental stage and sex, tissue type, and metathoracic scent gland for varying developmental stages and sexes) and one abiotic (RNA interference injection) conditions.  Four dedicated algorithms (ΔCt method, geNorm, BestKeeper and NormFinder) were used to analyze gene expression stability.  In addition, RefFinder provided an overall ranking of the stability/suitability of these candidates.  This study is the first to provide a comprehensive list of suitable reference genes for gene expression analyses in A. suturalis, which can serve to facilitate transcript expression study of related biological processes in this and related species.
 
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2. Toxicity and binding analyses of Bacillus thuringiensis toxin Vip3A in Cry1Ac-resistant and -susceptible strains of Helicoverpa armigera (Hübner)
ZHANG Qian, CHEN Li-zhen, LU Qiong, ZHANG Yan, LIANG Ge-mei
Journal of Integrative Agriculture    2015, 14 (2): 347-354.   DOI: 10.1016/S2095-3119(14)60770-X
摘要2171)      PDF    收藏
The Bacillus thuringiensis vegetative insecticidal protein, Vip3A, represents a new family of Bt toxin and is currently applied to commercial transgenic cotton. To determine whether the Cry1Ac-resistant Helicoverpa armigera is cross-resistant to Vip3Aa protein, insecticidal activities, proteolytic activations and binding properties of Vip3Aa toxin were investigated using Cry1Ac-susceptible (96S) and Cry1Ac-resistant H. armigera strain (Cry1Ac-R). The toxicity of Vip3Aa in Cry1Ac-R slightly reduced compared with 96S, the resistance ratio was only 1.7-fold. The digestion rate of full-length Vip3Aa by gut juice extracts from 96S was little faster than that from Cry1Ac-R. Surface plasmon resonance (SPR) showed there was no significant difference between the binding affinity of Vip3Aa and BBMVs between 96S and Cry1Ac-R strains, and there was no significant competitive binding between Vip3Aa and Cry1Ac in susceptible or resistant strains. So there had little cross-resistance between Vip3Aa and Cry1Ac,Vip3A+Cry proteins maybe the suitable pyramid strategy to control H. armigera in China in the future.
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