水稻丙酮酸激酶基因OsPK1的过表达转基因株系的获得与鉴定  

Generation and Characterization of Overexpression Lines of a Rice Pyruvate Kinase Gene OsPK1

作　　者：张艳[1,2] 庞金环[3] 吴家和[3] 何朝族[2,4] 

机构地区：[1]清华大学生命科学学院,北京100084 [2]清华大学深圳研究生院,广东深圳518055 [3]中国科学院微生物研究所,植物基因组国家重点实验室,北京100101 [4]海南大学海南省热带生物资源可持续利用重点实验室,海南海口570228

出　　处：《华北农学报》2012年第2期1-5,共5页Acta Agriculturae Boreali-Sinica

基　　金：海南省自然科学基金(30204)

摘　　要：利用农杆菌介导的转化方法研究OsPK1过表达对水稻的影响。将CaMV 35S启动子驱动的OsPK1的全长CDS序列导入到日本晴基因组中。选择3个过表达转基因系作为代表进行鉴定和分析。结果显示,过表达植株在株高、分蘖数、穗长和千粒重四方面接近野生型水稻,结实率略下降,但每穗饱满种子数明显增加。通过定量RT-PCR分析,发现有4个代谢酶在过表达植株嫩叶中表达存在着上调或下调。用GC-MS方法检测糖含量,显示OsPK1过表达株系中葡萄糖、果糖、半乳糖和蔗糖的含量与野生型差异不大。总之,对OsPK1过表达株系的鉴定,有助于更好了解OsPK1的功能,并且每穗饱满种子数明显增加具潜在的应用价值。We have reported that downregulation of OsPK1, a rice pyruvate kinase, causes dwarfism, reduced seed set and so on. Therefore,the aim in this study is to explore the effect of the overexpression of OsPK1 on rice. The construct containing a CaMV 35S-driven full-length CDS of OsPK1 was introduced into the genome of wild-type （WT） Nipponbare via an Agrobacterium-mediated method. Three overexpression lines were selected for further char- acterization and analysis. Statistics showed that these overexpression plants approach WT plants in plant height,tiller number, panicle length and thousand seed weight, while present slightly reduced seed set and significantly increased viable seed number per panicle. Quantitative RT-PCR results exhibited that the expressions of four metabolic en- zymes were downregulated or upregulated in the young leaves of overexpression plants. The sugar contents were measured using GC-MS, and the results indicated that the contents of glucose, fructose, galactose and sucrose in Os- PK1 overexpression plants showed few differences compared with those of WT plants. Together,the characterization of OsPK1 overexpression plants can help us better understand the role of OsPK1 ,and increased viable seed number per panicle should be a prospective application.

关 键 词：OsPK1 过表达 丙酮酸激酶 水稻 

分 类 号：Q78[生物学—分子生物学]