利用真空侵染法在紫花苜蓿中瞬时表达GUS基因

Expression of GUS Gene in Medicago sativa L. by Vacuum Infiltration

由于传统的苜蓿遗传转化体系存在周期长的局限性,本研究在紫花苜蓿(Medicago sativa L.)中建立并优化了一种快速、简单、高效的表达系统,即发芽种子真空侵染法。我们以发芽种子作为真空侵染材料,快速表达了β-葡萄糖苷酸酶(β-glucuronidase, GUS),并优化了这种表达系统的侵染条件。结果表明当真空侵染压强为0.08 Mpa,农杆菌重悬液浓度OD600为1.0时,被侵染的植物中GUS基因的表达效率较高。GUS化学组织染色检测和半定量RT-PCR检测结果表明,侵染后第14天GUS基因在苜蓿植物中被成功表达。本研究为植物生物反应器和紫花苜蓿(Medicago sativa L.)中基因功能分析提供了研究基础。

Due to the long cycle limitation of traditional genetic transformation system of alfalfa, we have developed and optimized a fast and simple expression system in Medicago sativa L., namely vacuum infiltration of germinated seeds. We used germinated seeds of alfalfa as the vacuum infiltration material, rapidly expressed the β-glucuronidase(β-glucuronidase, GUS), and optimized the infiltration conditions of this system. The results showed that the expression efficiency of GUS gene in the infected plants was higher when the vacuum pressure was 0.08 MPa and the Agrobacterium resuspension concentration OD600 was 1.0. The detection results of GUS chemical tissue staining and semi-quantitative RT-PCR showed that the GUS gene was successfully expressed on 14 days after infiltration in the alfalfa plants. This paper provided the basis research for plant bioreactor and study of gene function in Medicago sativa.