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1. JIA-2021-2149 基于手性异构体的茶毛虫高效性诱剂研发
LI Zhao-qun, YUAN Ting-ting, CUI Shao-wei, ZHAO Ying-jie, SHAO Yuan-hai, SHANG Jian-nong, LUO Zong-xiu, CAI Xiao-ming, BIAN Lei, CHEN Zong-mao
Journal of Integrative Agriculture    2023, 22 (1): 195-201.   DOI: 10.1016/j.jia.2022.08.113
摘要279)      PDF    收藏

研究茶毛虫主要性信息素组分的手性对映体和次要组分的田间引诱活性,明确其最佳配比,可为茶毛虫的高效性诱杀技术研发提供理论基础。本研究利用昆虫触角电位技术测定了茶毛虫雄蛾触角对主要性信息组分的两个手性对映体(S)-10,14-二甲基十五醇异丁酸酯和(R)-10,14-二甲基十五醇异丁酸酯及次要组分14-甲基十五醇异丁酸酯的电生理活性。通过田间诱捕试验研究3个单组分及其不同配比对茶毛虫雄蛾的引诱活性,并比较了其最优配比与商品化产品的田间引诱活性。(1(R)-10,14-二甲基十五醇异丁酸酯的电生理活性显著高于(S)-10,14-二甲基十五醇异丁酸酯,14-甲基十五醇异丁酸酯的电生理活性也具有一定的电生理活性;(2(R)-10,14-二甲基十五醇异丁酸酯的田间引诱活性显著高于(S)-10,14-二甲基十五醇异丁酸酯和外消旋体10,14-二甲基十五醇异丁酸酯;(314-甲基十五醇异丁酸酯虽无田间引诱活性,但其可以显著的提高其他两个组分的引诱活性,(4(R)-10,14-二甲基十五醇异丁酸酯的田间诱捕活性随浓度增加而增强,14-甲基十五醇异丁酸酯对 (R)-10,14-二甲基十五醇异丁酸酯诱捕活性的增强作用随浓度增加先增强后降低,性信息素配比以R)-10,14-二甲基十五醇异丁酸酯和4-甲基十五醇异丁酸酯含量分别为0.75mg 0.1mg时引诱活性最强,显著优于现有商品化产品。以往的研究报道显示,中国种群茶毛虫性信息素仅有10,14-二甲基十五醇异丁酸酯一个组分,且该组分的两个手性对映体田间引诱活性差异不显著。本研究通过系统的研究,明确了(R)-10,14-二甲基十五醇异丁酸酯是茶毛虫的主要性信息素组分,14-甲基十五醇异丁酸酯可能是茶毛虫次要组分。在此基础上,获得了一个最佳的茶毛虫性信息素配比,可用于研发高效茶毛虫性诱杀技术

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2. Validation of Reference Genes for Quantitative Real-Time PCR in Laodelphax striatellus
HE Xiu-ting, LIU Cheng-cheng, LI Zhao-qun, ZHANG Zan, LI Guo-qing, LI Fei , DONG Shuang-
Journal of Integrative Agriculture    2014, 13 (4): 811-818.   DOI: 10.1016/S2095-3119(13)60515-8
摘要2579)      PDF    收藏
The normalization of quantitative real-time PCR (qPCR) is important to obtain accurate gene expression data, and the most common method for qPCR normalization is to use reference genes. However, reference genes can be regulated under different conditions. qPCR has recently been used for gene expression study in Laodelphax striatellus, but there is no study on validation of the reference genes. In this study, five new housekeeping genes (LstrTUB1, LstrTUB2, LstrTUB3, LstrARF and LstrRPL9) in L. striatellus were cloned and deposited in the GenBank with accession numbers of JF728809, JF728810, JF728811, JF728807 and JF728806, respectively. Furthermore, mRNA expressions of the five genes and β-actin were measured by qPCR with insect samples of different instar at nymph stage, and the expression stabilities were determined by the software geNorm and NormFinder. As a result, ARF and RPL9 were consistently more stable than β-actin, while three TUB genes were less stable than β-actin. To determine the optimal number of reference genes used in qPCR, a pairwise variations analysis by geNorm indicated that two references ARF and RPL9 were required to obtain the accurate quantification. These results were further confirmed by the validation qPCR experiment with chitinase gene as the target gene, in which the standard error of the mRNA quantification by using binary reference ARF-RPL9 was much lower than those by ARF, RPL9 or β-actin alone. Taken together, our study suggested that the combination of ARF-RPL9 could replace β-actin as the reference genes for qPCR in L. striatellus.
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