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赤红壤地区森林土壤球囊霉素相关蛋白测定方法

张静1,2,3,唐旭利1,3**,郑克举1,2,3,童琳1,2,3,陈红锋1   

  1. (1中国科学院华南植物园,广州 510650;2中国科学院大学,北京 100049;3中国科学院退化生态系统植被恢复与管理重点实验室,广州 510650)
  • 出版日期:2014-01-10 发布日期:2014-01-10

An improved procedure for glomalin-related soil protein measurement in subtropical forest.

ZHANG Jing1,2,3, TANG Xu-li1,3**, ZHENG Ke-ju1,2,3, TONG Lin1,2,3, CHEN Hong-feng1   

  1. (1South China Botanical Garden, Chinese Academy of Sciences, Guangzhou 510650, China; 2University of Chinese Academy of Sciences, Beijing 100049, China; 3Key Laboratory of Vegetation Restoration and Management of Degraded Ecosystem, Chinese Academy of Sciences, Guangzhou 510650, China)
  • Online:2014-01-10 Published:2014-01-10

摘要: 球囊霉素是丛枝菌根真菌菌丝产生的一种糖蛋白,被证明具有改善土壤结构、固定土壤中重金属、保护有机碳等重要的生态功能。球囊霉素目前尚无法纯化,一般通过测定球囊霉素相关蛋白(GRSP)来对其定量。由于GRSP测定方法的非专一性,目前尚无适宜于各种陆地生态系统的标准测定方法。本文针对GRSP测定结果可能受到提取时间、离心力以及测定前再次离心时离心力的大小等因素的影响,设置对比试验,对亚热带赤红壤地区土壤GRSP提取测定,以期获取适合于该地区的GRSP测定方法。结果表明:当提取时离心时间为10 min,10000×g的离心力提取的易提取GRSP(EE-GRSP)是5000×g提取的1.15~1.82倍,但离心力大小对总GRSP(T-GRSP)影响不显著;当提取时离心力为5000×g时,离心时间由10 min延长至15 min能够显著提高EE-GRSP含量;测定之前对上清液再次离心,若使用较大的离心力(≥10000×g),将显著影响GRSP测定值;经过6次连续提取,能够提取T-GRSP理论最大含量的97.7%~99.8%;因此,在亚热带酸性赤红壤森林生态系统中,GRSP的提取建议采用10000×g 10 min的离心力和离心时间组合,测定前对上清液再次离心时,应以≥10000×g的离心力为宜,研究结果为该地区GRSP的标准化测定提供了理论依据。

关键词: 温度, 相对湿度, 形状特征, 城市湿地

Abstract: Glomalin as a recalcitrant glycoprotein produced by arbuscular mycorrhizal fungi is proved to improve soil structure, fix heavy metals in soils, and be a substantial contributor to soil carbon. At present, glomalin cannot be directly purified but can be quantified by measuring the content of glomalin-related soil protein (GRSP) in soil. Currently no standard procedure for GRSP measurement is available for all terrestrial ecosystems because of the non-specific nature of GRSP measurement. To investigate the possible influences of extraction time, centrifugal force, and centrifugal force before colorimetric test on GRSP measurement, we carried out a comparative study using latosolic red soils collected from a subtropical forest.  Higher centrifugal force extracted more easily extractable GRSP (EE-GRSP) for a given centrifuge time. Under 10 minutes of extraction, EEGRSP extracted with centrifugal force at 10000×g was 1.15-1.82 times more than at 5000×g, but the effect of centrifugal force on T-GRSP was not significantly different at α=0.05 level. Extending centrifugal time increased EE-GRSP concentration for a given centrifugal force. With the centrifugal force at 5000×g, EE-GRSP content increased significantly with the increase of centrifugation time from 10 minutes to 15 minutes. Centrifugation of the supernate with higher centrifugal force (≥10000×g) before colorimetric test significantly influenced the measurement of GRSP. The T-GRSP value reached 97.7%-99.8% of the theoretical maximum after 6 times of repeated extraction. Based on this study, the procedure for GRSP measurement for latosolic red soils in subtropical forests is recommended as centrifugation of soil solution at 10000×g for 10 minutes, and centrifugation of the supernate at 10000×g before colorimetric test.

Key words: relative humidity., temperature, urban wetland, shape feature