摘要： 为初步探讨V-ATPase c亚基VHA-c1基因在植物生长发育及信号转导中的作用，通过农杆菌介导，将设计的dsDNA片段整合到拟南芥基因组上，筛选获得了能够特异沉默VHA-c1基因的转基因株系c1-1，并对其进行NaCl、ABA和6%葡萄糖胁迫处理。结果表明：在NaCl处理后，沉默株系c1-1的主根伸长量和种子萌发率都有明显的降低，且在浓度为50，100 mmol/L NaCl胁迫处理下，转基因株系c1-1的主根伸长量被显著抑制。在特定浓度的ABA和6%葡萄糖处理后，转基因株系c1-1的主根伸长量和种子萌发率也都有一定程度的抑制。然而，在4，8 μmol/L ABA处理后，野生株系的主根伸长量被显著抑制，与转基因株系c1-1的主根伸长量被抑制效果相比达到了显著水平。沉默VHA-c1基因后，降低了拟南芥对NaCl的耐受性，影响了其生长发育，然而对ABA和葡糖糖的抑制作用却表现为很不敏感，这可能是因为H⁺-ATPase在盐胁迫信号通路和ABA信号通路中起着不同的调控功能。

关键词: 液泡ATP酶, 基因干扰技术, 非生物胁迫, 拟南芥

Abstract: The vacuolar type H⁺-ATPase(VHA)is a multi-subunit endomembrane proton pump and plays a important role in plant adaptation and resistance to abiotic stresses. In order to explore the function of VHA subunit c gene(VHA-c1)in plant growth and signal transduction, the designed double strand DNA fragment was integrated into the genome of Arabidopsis thaliana by Agrobacterium mediation, and then a transgenic line c1-1 was screened to specifically silence the coding gene of VHA-c1 subunit. The results showed that the relative root length and the germination rate of seeds in c1-1 line decreased compared with the wild strain under the stress of NaCl. Especially, under the stress of 50,100 mmol/L NaCl, the relative root length of c1-1 line was suppressed sharply. Under the stress of ABA or glucose, the relative root length of c1-1 line and wild strain was suppressed. However, the relative root length of wild strain decreased more obviously than c1-1 line at 4,8 μmol/L ABA treatment. It suggested that the silenced VHA-c1 gene significantly reduced the tolerance of Arabidopsis to NaCl, and was insensitive to ABA and glucose stresses. This might be due to the different regulatory functions of H⁺-ATPase in salt stress and ABA signaling pathways.

Key words: Vacuolar type H⁺-ATPase, RNA interference technology, Abiotic stresses, Arabidopsis thaliana

中图分类号: 

• Q945.78