摘要
【目的】卡尔文循环是植物光合作用中极为重要的生理过程,对植物的生长发育具有显著影响。前期研究表明,高光合速率的速生欧美杨的景天庚酮糖-1,7-二磷酸酯酶(SBPase)基因表达水平在速生期显著上调,预示该基因在光合碳固定过程中可能起着关键作用。【方法】为进一步解析SBPase在木本植物光合速率和生长发育中的作用,本文从速生欧美杨品系NE19中克隆得到了PdSBPase基因,并构建35S:PdSBP:GFP表达载体,采用农杆菌花序侵染法转化拟南芥,通过抗生素筛选,PCR鉴定和组织定位等多种方式鉴定并成功得到了超表达PdSBPase拟南芥株系。【结果】在正常生长状态下,超表达植株的叶面积、根长、株高都优于野生型和突变体,其中叶面积是野生型的1.79倍,根长是野生型的1.93倍,而突变体表现为植株矮化,叶子明显发黄短小,叶绿素含量低于野生型株系。转基因株系SBPase酶活是野生型1.4倍,是突变体的1.9倍,RuBP产量以及淀粉含量均要高于野生型和突变体株系,RuBP产量分别是野生型和突变体的1.37和1.76倍,转基因株系的淀粉含量达到了50.26μg/g,而突变体的淀粉含量未检出。【结论】这些结果说明,PdSBPase对RuBP的形成和淀粉等多糖的合成起到关键作用,能促进植物积累更多的碳水化合物,进而正向调控植物的光合能力。
[Objective]Calvin cycle plays an important role in the photosyntheic process and strongly impacts the growth and development of plants. Previous studies have showed that expression level of sedoheptulose-1,7-bisphosphatase（ SBPase） gene is significantly up-regulated in the fast-growing hybrid poplar with the high photosynthetic rate in fast-growth phase,suggesting that the gene may plays a key role in the photosynthetic carbon fixation. [Method]In order to further analyze the function of SBPase in photosynthetic efficiency and plant growth and development in woody plants,PdSBP gene was cloned from fast-growing hybrid Populus NE-19（ Populus nigra ×（ Populus deltoids × Populus nigra））. The 35 S：PdSBP： GFP expression vector was constructed and transformed into Arabidopsis thaliana by Agrobacterium tumefaciens inflorescence infection. We used antibiotic selection,PCR identification and subcellular localization to successfully achieve the overexpressing PdSBPase Arabidopsis. [Result]Undernormal growth conditions,the leaf area,root length,and plant height of overexpressing plants were all better than those of wild type and mutant. Among them,the leaf area of transgenic plants was 1. 79 times of the wild type and the root length was 1. 93 times of the wild type. In comparison,the mutant had an obvious dwarfing phenomenon,with yellow and short leaves. Furthermore,chlorophyll content was lower than other genotypes. In addition,the SBPase activity of transgenic lines was 1. 4 times of wild type,1. 9 times of the mutant. The RuBP yield and starch content were also higher than those of wild type and the mutant. The yield of RuBP in transgenic plants was 1. 37 and 1. 76 times of the wild type and the mutant,respectively. The starch content of the transgenic lines reached 50. 26 μg/g,while that of the mutants was not detected. [Conclusion] These results indicate that PdSBPase plays a vital role in the formation of RuBP and starch and other polysaccharides synthesis,and can promote plants to accumulate more carbohydrates and then positively regulate the photosynthetic capacity of plants.
作者
王俊秀
周扬颜
韩潇
安轶
郭惠红
夏新莉
尹伟伦
刘超
Wang Junxiu, Zhou Yangyan, Han Xiao, An Yi, Guo Huihong, Xia Xinli, Yin Weilun, Liu Chao(National Engineering Laboratory for Tree Breeding, Beijing Forestry University, Beijing 100083, Chin)
出处
《北京林业大学学报》
CAS
CSCD
北大核心
2018年第3期26-33,共8页
Journal of Beijing Forestry University
基金
国家重点研发计划课题(2016YFD0600403)
国家自然科学基金项目(31570308
31600484
31770649)
