Key Laboratory of Molecular and Cytogenetics, College of Life Science and Technology, Harbin Normal University, Harbin, 150025, Heilongjiang, China.
Ecotoxicology. 2014 May;23(4):718-25. doi: 10.1007/s10646-014-1185-y. Epub 2014 Jan 16.
Green fluorescent protein (GFP) is one of the most widely studied and exploited proteins in biochemistry, and has many applications as a marker, especially in plant transformation system. Although a number of studies have been conducted to assess the toxify of this protein to specific organisms, little is known about GFP on rhizosphere microbial community, which is regarded as good indicator for environmental risk assessment. Chloroplast genetic engineering has shown superiority over traditional nuclear genetic engineering, and has been used in many aspects of plant genetic engineering. High levels of chloroplast-based protein accumulation make this technology as an ideal strategy to evaluate biosafety of transgenes. In the present study, the effects of field-released GFP transplastomic tobacco (Nicotiana tabacum) on rhizosphere microbes over a whole growth cycle were investigated by using both culture-dependent and culture-independent methods. Compared to wild-type control, transplastomic tobacco had no significant influence on the microbial population at the seedling, vegetative, flowering and senescing stages. However, developmental stages had more influence than ecotypes (GFP-transformed and wild-type). This was confirmed by colony forming unit, Biolog Eco(TM) and PCR-DGGE analysis. Thus, these results suggest chloroplast transformation with a GFP reporter gene has no significant influence on rhizosphere microbial community, and will be potential platform for plant biotechnology in future.
绿色荧光蛋白（GFP）是生物化学中研究和应用最广泛的蛋白质之一，作为标记物有许多应用，特别是在植物转化系统中。尽管已经进行了许多研究来评估该蛋白对特定生物体的毒性，但对于根际微生物群落中 GFP 的了解甚少，而根际微生物群落被认为是环境风险评估的良好指标。叶绿体遗传工程优于传统的核遗传工程，并已被用于植物遗传工程的许多方面。基于叶绿体的高水平蛋白积累使这项技术成为评估转基因生物安全性的理想策略。在本研究中，采用培养依赖和培养非依赖的方法，研究了田间释放的 GFP 叶绿体转化烟草（Nicotiana tabacum）对整个生长周期根际微生物的影响。与野生型对照相比，叶绿体转化烟草对幼苗、营养生长、开花和衰老阶段的微生物种群没有显著影响。然而，发育阶段比生态型（GFP 转化和野生型）的影响更大。这一点通过平板菌落形成单位计数、Biolog Eco（TM）和 PCR-DGGE 分析得到了证实。因此，这些结果表明，用 GFP 报告基因进行叶绿体转化对根际微生物群落没有显著影响，并且将来可能成为植物生物技术的平台。
