| DNA-SIP鉴定甘蔗//大豆间作土壤15N-DNA富集位置的氮循环功能基因qPCR方法 |
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| 引用本文： | 苟永刚,余玲玲,许霞,王建武.DNA-SIP鉴定甘蔗//大豆间作土壤15N-DNA富集位置的氮循环功能基因qPCR方法[J].农业环境科学学报,2019,38(1):140-147. |
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| 作者姓名： | 苟永刚 余玲玲 许霞 王建武 |
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| 作者单位： | 华南农业大学热带亚热带生态研究所, 广州 510642;农业农村部华南热带农业环境重点实验室, 广州 510642;华南农业大学资源环境学院生态学系, 广州 510642,华南农业大学热带亚热带生态研究所, 广州 510642;农业农村部华南热带农业环境重点实验室, 广州 510642;华南农业大学资源环境学院生态学系, 广州 510642,华南农业大学热带亚热带生态研究所, 广州 510642;农业农村部华南热带农业环境重点实验室, 广州 510642;华南农业大学资源环境学院生态学系, 广州 510642,华南农业大学热带亚热带生态研究所, 广州 510642;农业农村部华南热带农业环境重点实验室, 广州 510642;华南农业大学资源环境学院生态学系, 广州 510642 |
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| 基金项目： | 国家自然科学基金项目（31600348） |
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| 摘 要： | 为筛选有效鉴定甘蔗//大豆间作系统的稳定性同位素核酸探针技术(DNA-SIP)中超高速离心后15N-DNA富集位置的指示功能基因,利用实时荧光定量PCR技术(qPCR),检测6个氮素循环功能基因在不同浮力密度离心液DNA中的相对丰度分布,通过对氮素循环功能基因相对丰度作图分析,nifH和amoA基因在甘蔗//大豆间作和大豆单作种植模式中15N标记组与对照组基因丰度峰发生偏移,chiA基因丰度峰仅在大豆单作种植模式下存在偏移,而nirS、nirK、nosZ等3个基因的丰度峰值在两种种植模式下均不发生偏移。结果表明nifH和amoA基因可作为指示基因,能够有效鉴定甘蔗//大豆间作系统中DNA-SIP技术15N-DNA位置。
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| 关 键 词： | 甘蔗//大豆间作 稳定性同位素核酸探针 荧光定量PCR 功能基因 |
| 收稿时间： | 2018/3/15 0:00:00 |
Identification of 15N-DNA enrichment sites in DNA-SIP to reveal functional genes by qPCR from sugarcanesoybean intercropping soil |
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| GOU Yong-gang,YU Ling-ling,XU Xia and WANG Jian-wu.Identification of 15N-DNA enrichment sites in DNA-SIP to reveal functional genes by qPCR from sugarcanesoybean intercropping soil[J].Journal of Agro-Environment Science( J. Agro-Environ. Sci.),2019,38(1):140-147. |
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| Authors: | GOU Yong-gang YU Ling-ling XU Xia and WANG Jian-wu |
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| Institution: | Institute of Tropical and Subtropical Ecology, South China Agricultural University, Guangzhou 510642, China;Key Laboratory of Agroenvironment in the Tropics, Ministry of Agriculture and Rural Affairs, Guangzhou 510642, China;Department of Ecology, College of Natural Resources and Environment, South China Agricultural University, Guangzhou 510642, China,Institute of Tropical and Subtropical Ecology, South China Agricultural University, Guangzhou 510642, China;Key Laboratory of Agroenvironment in the Tropics, Ministry of Agriculture and Rural Affairs, Guangzhou 510642, China;Department of Ecology, College of Natural Resources and Environment, South China Agricultural University, Guangzhou 510642, China,Institute of Tropical and Subtropical Ecology, South China Agricultural University, Guangzhou 510642, China;Key Laboratory of Agroenvironment in the Tropics, Ministry of Agriculture and Rural Affairs, Guangzhou 510642, China;Department of Ecology, College of Natural Resources and Environment, South China Agricultural University, Guangzhou 510642, China and Institute of Tropical and Subtropical Ecology, South China Agricultural University, Guangzhou 510642, China;Key Laboratory of Agroenvironment in the Tropics, Ministry of Agriculture and Rural Affairs, Guangzhou 510642, China;Department of Ecology, College of Natural Resources and Environment, South China Agricultural University, Guangzhou 510642, China |
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| Abstract: | Using DNA stable isotope probes(DNA-SIPs) is a reliable, new technique for studying the nitrogen cycle. To identify indicator function genes of 15N-DNA enrichment in ultra-high-speed centrifugation in a sugarcane-soybean intercropping system for DNA-SIPs, the relative abundance distributions of six nitrogen cycling functional genes in the DNA of different buoyant density centrifugation fluids were detected via real-time PCR(qPCR). Through the analysis of the relative abundance of nitrogen cycling functional genes, the gene abundance peaks of the nifH and amoA genes in sugarcane-soybean intercropping and soybean monoculture were shifted in the 15N marker group and the control group, the gene abundance peaks of chiA were only shifted in soybean monocropping mode, and the abundance peaks of nirS, nirK, and nosZ did not shift with either planting pattern. The results showed that the nifH and amoA genes could be used as indicator genes to effectively identify DNA-SIP technology 15N-DNA positions in sugarcane-soybean intercropping systems. |
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| Keywords: | sugarcane-soybean intercropping stable isotope probing real-time PCR functional genes |
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