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    继代培养14年的转AhDREB1基因毛白杨环境释放的安全性评价

    Safety evaluation of environmental release of transgenic Populus tomentosa with AhDREB1 gene after 14 years of subculture

    • 摘要:
      目的 本研究以继代培养14年,而后移栽到天津和河北大田生长6年的转 AhDREB1 基因毛白杨为研究对象,从生长适应性、基因表达稳定性、基因水平转移、对土壤环境影响以及根际土浸提液的化感作用等方面,系统评价转基因毛白杨经多年继代培养后又于田间释放后的稳定性与安全性,旨在提供转基因林木田间试验长期监测数据,为转基因林木的后续推广应用提供科学支持。
      方法 通过测量转基因毛白杨在天津滨海和河北沧州盐碱地试验林的树高、胸径、材积等生长指标来分析其生长状况,提取叶片和土壤及微生物总 DNA后运用 PCR 技术测定外源基因在转基因毛白杨中的表达稳定性以及探究外源基因对土壤环境的影响,采用稀释平板计数法解析不同月份土壤微生物数量,利用根际土浸提液来检测转基因杨树根系分泌物对白菜种子胚根、胚轴生长的化感效应。
      结果 (1)转AhDREB1基因毛白杨在天津滨海和河北沧州盐碱地的生长情况皆优于非转基因株系。(2)外源基因 AhDREB1 在继代培养14年又田间生长6年的转基因毛白杨中仍稳定存在,未检测到外源基因向土壤及土壤微生物转移。(3)不同季节,转AhDREB1基因毛白杨与对照毛白杨的土壤微生物数量均无显著差异,其根际土浸提液对白菜种子胚根和胚轴的生长影响均无显著差异。
      结论 在盐碱地条件下,转AhDREB1基因毛白杨生长适应性显著增强,且该基因在多年生转基因植株内稳定表达,未发现对环境产生生态安全隐患问题。本研究结果将为AhDREB1转基因株系在盐碱环境下的后续推广应用提供科学依据。

       

      Abstract:
      Objective This study used transgenic plants that had been subcultured for 14 years and subsequently grown in field conditions in Tianjin and Hebei of northern China for 6 years as research materials. The assessment covered multiple aspects including growth adaptability, gene expression stability, horizontal gene transfer potential, impact on soil environment, and allelopathic effects of rhizosphere soil extracts. The study systematically evaluated the stability and safety of transgenic P. tomentosa for AhDREB1 gene after long-term subculture and field release. The aim of this research was to provide long-term monitoring data from field trials of transgenic trees, thereby offering scientific support for the subsequent application and promotion of transgenic woody plants.
      Method The growth status of transgenic P. tomentosa was analyzed by measuring its growth indicators such as tree height, DBH, and volume in the experimental forests on saline-alkali research sites in Tianjin Binhai and Hebei Cangzhou. After extracting total DNA from leaves, soil, and microorganisms, PCR technology was used to determine expression stability of exogenous genes in transgenic P. tomentosa and to explore the impact of exogenous genes on soil environment. The dilution plate counting method was adopted to analyze the quantity of soil microorganisms in different months. The allelopathic effect of root exudates from transgenic poplars on the growth of radicle and hypocotyl of Chinese cabbage (Brassica pekinensis) seeds was detected using rhizosphere soil extracts.
      Result (1) The growth of transgenic P. tomentosa for AhDREB1 gene in the Tianjin Binhai and Hebei Cangzhou saline-alkali research sites was better than that of the non-transgenic lines. (2) The exogenous gene AhDREB1 still stably existed in the transgenic poplar through 14 years of subculturing and 6 years of field cultivation. No transfer of exogenous gene to soil and soil microorganisms was detected. (3) In different seasons, there was no significant difference in the quantity of soil microorganisms between the transgenic P. tomentosa for AhDREB1 gene and control. Additionally, the extracts of their rhizosphere soils showed no significant differences in their effects on the growth of radicles and hypocotyls of Chinese cabbage seeds.
      Conclusion Under saline-alkali soil conditions, transgenic P. tomentosa for AhDREB1 gene exhibites significantly enhanced growth adaptability compared with non-transgenic P. tomentosa. Moreover, the AhDREB1 gene was stably expressed in perennial plants, and no potential ecological security risks to the environment caused by P. tomentosa transgenic for the AhDREB1 gene were observed. The results of this study will lay a scientific foundation for promotion and application of this transgenic line in saline-alkali environments.

       

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