《植物生理学报》 2013, 49(12): 1447-1453

间接竞争ELISA测定花生植株内源水杨酸含量

鄢洪海^1,*, 王学武^2,*, 张茹琴¹, 况川¹, 迟玉成³, 夏淑春^1,**

¹青岛农业大学农学与植物保护学院, 山东青岛266109; ²青岛市植物保护站, 山东青岛266100; ³山东省花生研究所, 山东青岛266101

通信作者：鄢洪海；E-mail: xiashuchun@163.com；Tel: 0532-88030496

摘　要：

以SA-N=CH-(CH₂)₃-CH=N-OVA为包被抗原, 利用SA-NH-CH₂-NH-BSA为免疫原产生的抗体, 来建立能定量测定花生组织中游离水杨酸(SA)的间接竞争ELISA工作程序, 并明确该方法的工作条件以及基本参数。结果表明: 该方法对SA标准品的最低检测量为10 ng·mL^-1, 线性检测范围为10 ng·mL^-1~10 μg·mL^-1。在该范围内, 标准曲线的批内和批间变异系数分别为1.90%和6.81%; 花生叶片和根组织中添加SA的平均回收率分别为93.9%和91.0%。

关键词：花生; 内源水杨酸; 间接竞争ELISA

收稿：2013-06-27   修定：2013-09-27

资助：山东省科技发展项目(2009GG10009022)、山东省自然科学基金项目(ZR2011CL005)和山东省“泰山学者”建设工程专项经费(BS2009NY040)

An Indirect Competitive ELISA Determination to Detect Salicylic Acid in Peanut Plant

YAN Hong-Hai^1,*, WANG Xue-Wu^2,*, ZHANG Ru-Qin¹, KUANG Chuan¹, CHI Yu-Cheng³, XIA Shu-Chun^1,**

¹College of Agronomy and Plant Protection, Qingdao Agricultural University, Qingdao, Shandong 266109, China; ²Qingdao Plant Protection Station, Qingdao, Shandong 266100, China; ³Peanut Research Institute in Shandong Province, Qingdao, Shandong 266101, China

Corresponding author: YAN Hong-Hai; E-mail: xiashuchun@163.com; Tel: 0532-88030496

Abstract:

SA-N=CH-(CH₂)₃-CH=N-OVA was used as coating antigen, and SA-NH-CH₂-NH-BSA was used as immunogen to induce antibodies to establish a procedure that could quantitatively determine free salicylic acid (SA) in peanut plants, and clear the working conditions and basic parameters in the procedure. The results indicated that, using the procedure, the minimum detectable content of SA in peanut plants was 10 ng·mL^-1, the linear detectable content range from 10 ng·mL^-1 to 10 μg·mL^-1. Within this range, the intra- and inter-assay coefficients of standard curve were 1.90% and 6.81%, respectively; the average recovery of added SA in peanut leave and root tissues were 93.9% and 91.0%, respectively.

Key words: Arachis hypogaea; endogenesis salicylic acid; indirect competitive ELISA