Over-expression of RPEase gene promotes the growth and development of Arabidopsis thaliana
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摘要: 卡尔文循环是光合作用CO2同化的重要途径,在植物生长发育过程中起着重要作用。磷酸核酮糖3-差向异构酶(RPEase:EC 5.1.3.1)是卡尔文循环再生阶段的一种重要酶类。本文从速生欧美杂交黑杨NE-19中克隆得到RPEase基因,构建植物表达载体,利用农杆菌花序侵染法转化野生型和突变体拟南芥,通过普通PCR检测和绿色荧光蛋白(GFP)观测进一步鉴定得到CaMV35S:PdRPE:GFP超表达株系,然后对野生型、超表达株系、突变体和回补株系的生理指标进行测定。结果显示,超表达株系RPEase活性显著升高(P<0.05)。在正常浇水的生长条件下,超表达株系相比于其他3个株系(野生型、突变体、回补株系),气孔数目减少,气孔变大,提高了植物的水分利用效率以及净光合速率,使得超表达株系有更好的生长优势,积累更多的淀粉。在10d的短期干旱条件下,超表达株系的净光合速率和水分利用效率依然显著高于其他3个株系(P<0.01)。因此,研究表明超表达RPEase基因会提高植物生物量的积累以及对短期干旱的抵抗能力。Abstract: The Calvin cycle is an irreplaceable pathway for photosynthetic organisms to assimilate CO2 from the air and therefore plays a crucial role in plant growth and development. The ribulose-phosphate 3-epimerase (RPEase: EC 5.1.3.1) is one of the integral enzymes in the Calvin cycle regeneration phase. In our study PdRPE gene was cloned from fast-growing hybrid poplar NE-19 (Populus nigra × (Populus deltoids × Populus nigra)), and then PdRPE was transformed into Arabidopsis thaliana by the floral dip method. Using PCR and Green Fluorescent Protein (GFP) detection methods, we obtained the CaMV35S:PdRPE:GFP transgenic plants. Then we validated the growth status and physiological indexes of wild type (Col-0), overexpressing (OxPdRPE), mutant (rpe) and complementary (rpe/PdRPE) lines. The results indicated that in overexpressing lines RPEase activity increased significantly (P<0.05). When well-watered, the overexpressing lines had lower leaf stomatal density, larger stomatal size, improved water use efficiency and higher net photosynthetic rate, showing a better growth condition and more starch accumulation. When exposed to 10 days of short-term drought, the net photosynthetic rate and water use efficiency of overexpressing lines were still significantly higher than those of the other three strains (P<0.01). Therefore, we reach the conclusion that over-expression of RPEase gene can promote the plant growth and development and enhance the resistance of short-duration drought in Arabidopsis thaliana.
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Keywords:
- poplar /
- Calvin cycle /
- RPEase /
- growth /
- development /
- water use efficiency
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