农业生物技术学报2012，Vol.20Issue(4)：347-355,9.DOI:10.3969/j.issn.1674-7968.2012.04.002
甘蔗果糖-6-磷酸,2-激酶/果糖-2,6-二磷酸酯酶基因(F2KP)的克隆及其功能研究
Cloning and Function Analysis of the Fructose-6-phosphate,2-kinase/fructose-2,6-bisphosphatase Gene (F2KP) from Sugarcane (Saccharum officinarum L.)
摘要
Abstract
Fructose- 6 -phosphate,2-kinase/fructose-2,6-bisphosphatase (F2KP), which is comfirmed to be a bifunctional enzyme, is a key regulatory enzyme involved in carbon accumulation and partitioning processes. In the previous study, F2KP with both kinase and phosphatase domain named SoF2KP-L was obtained from the leaf of sugarcane. In this research, different sizes of F2KP homologous fragments from the stem of sugarcane (Saccharum qfficinarum L.) were amplified and named SoF2KP-Sl, SoF2KP-S2 and SoF2KP-S3, respectively. Based on the bioinfomatical analysis, the putative protein SoF2KP-Sl, SoF2KP-S2 and SoF2KP-S3 were incomplete respectively with part of kinase domain, phosphatase domain and only several ammo acid. Therefore, SoF2KP-L with complete kinase/phosphatase catalytic domains was inserted into plant expression plasmid under the Cauliflower mosaic virus 35S promoter, then transferred into tobacco (Nicotiana tabacum) plants by Agrobacierium-mediated transformation. RT-PCR (Reverse transcription PCR) analysis confirmed that SoF2KP-L was successfully transcripted in mature leaf. The changing rates of soluble sugar/starch, reducing sugar/starch and sucrose/starch indicated that carbohydrate level and carbohydrate partitioning of the transgenic plants had been changed. These results indicate that there may be different transcripts existing in sugarcane and SoF2KP-L regulates the carbon portioning in sugarcane as F2KP in other plants. This stuty will provide theory basis for further study on the biological function of F2KP from sugarcane and for its potential application in sugarcane molecular breeding.关键词
甘蔗/F2KP/转基因/烟草/转录产物Key words
Sugarcane/ F2KP/ Transgenic plants/ Tobacco/ Transcripts引用本文复制引用
何炜,周平,张建福,逯平杰,苏轶,叶冰莹,雷美华,陈由强,陈如凯..甘蔗果糖-6-磷酸,2-激酶/果糖-2,6-二磷酸酯酶基因(F2KP)的克隆及其功能研究[J].农业生物技术学报,2012,20(4):347-355,9.基金项目
农业部现代农业产业技术体系建设专项(No.CARS-20-4-4)和国家自然科学基金(No.30871575) （No.CARS-20-4-4）
