RISA、DGGE和2D-PAGE技术对大豆根际土壤细菌多样性分析的比较

Bacterial diversity in rhizosphere soil of soybean:A comparison of RISA,DGGE,and 2D-PAGE techniques

采用基于PCR扩增的核糖体间隔区分析(RISA)、变性梯度凝胶电泳(DGGE)和双向电泳(2D-PAGE)3种分子生态学技术对大豆根际土壤细菌多样性比对分析。结果表明:2D-PAGE技术得到的土壤细菌多样性(基因点)最丰富,其次为DGGE技术(基因片段),RISA技术(基因片段)最低。RISA技术得到的条带数最少,但结果稳定性较高,并且实验操作比较简单;DGGE技术得到的条带数较多,具有较高的精度,但误差来源也最多;2D-PAGE技术作为一种新颖的土壤微生物研究方法,可以很好地解决其他2种技术分辨率低的缺点,能够获得丰富的土壤细菌多样性信息,在土壤微生物生态学研究中发挥重要作用,但与前2种技术相比,其操作比较复杂,条件要求相对比较严格。尽管存在不足,2D-PAGE技术在土壤微生物生态学研究领域中已显示出潜在的优势。

The PCR-based RISA, DGGE, and 2D-PAGE were adopted to analyze the bacterial diversity in rhizosphere soil of soybean. Most abundant soil bacterial diversity was acquired by 2D-PAGE （gene spots）, followed by DGGE （gene bands）, and by RISA （gene bands）. The RISA gave the fewest bands, but the result was more stable and the experimental operation was more easy; DGGE obtained more bands and showed good precision, but had more error sources; while 2D-PAGE as a novel soil microbial research method could overcome the low-resolution problem of the other two methods, achieve abundant information of soil bacterial diversity, and play an important role in the research of soil microbial ecology. However, the operation of 2D- PAGE was more complicated and required strict conditions, compared with RISA and DGGE. In spite of these disadvantages, 2D-PAGE had shown potential advantages in the research field of soil microbial ecology.