反义抑制水稻蔗糖转运蛋白基因(OsSUT5)的表达降低其愈伤组织诱导和植株再生频率  

Antisense Suppression Expression of Rice(Oryza sativa L.) Sucrose Transporter Gene(OsSUT5) Leads to Reducing Callus Induction and Plantlet Regeneration

作　　者：张武君[1,2] 管其龙 付艳萍[1] 苏军[1] 

机构地区：[1]福建省农业遗传工程重点实验室/福建省农业科学院生物技术研究所,福州350003 [2]福建农林大学生命科学学院,福州350002

出　　处：《农业生物技术学报》2014年第7期825-831,共7页Journal of Agricultural Biotechnology

基　　金：福建省自然科学基金资助项目(No.2011J01105)

摘　　要：蔗糖转运蛋白(sugar transporters,SUT)是一类介导蔗糖跨膜运输的蛋白,在植物的生长发育过程中起重要作用。水稻(Oryza sativa L.)SUT家族中有5个蔗糖转运蛋白,其中OsSUT3、OsSUT4和OsSUT5的大部分功能还未揭晓。本研究以转反义OsSUT5基因水稻为实验材料,通过抑制OsSUT5基因的表达,比较愈伤组织增殖率和植株分化率,分析反义抑制OsSUT5基因表达对愈伤组织诱导和分化能力的影响。结果显示,反义抑制OsSUT5的水稻,盾片膨大以及愈伤组织的诱导和增殖率显著下降(P<0.05),第1代愈伤组织干重为0.028 80 g/皿,第2代为0.119 93 g/皿,显著低于野生型水稻的0.058 70 g/皿和0.174 55 g/皿(P<0.05);继代培养5代之后,转基因和野生型的水稻愈伤组织鲜重增加没有显著差异,但转基因水稻的愈伤组织活力下降、水渍状明显,绿苗分化率为65%,显著低于野生型水稻的82.5%(P<0.05);对愈伤组织的OsSUTs基因定量分析显示,与野生型水稻相比,转基因水稻的OsSUT5和OsSUT1基因表达量下降近1倍,而OsSUT2和OsSUT4基因表达差异不显著(P>0.05)。研究结果提示,OsSUT5基因参与水稻组织培养过程中外植体对蔗糖的吸收和转运,为深入解析OsSUT5基因的功能提供参考资料。Sugar transporters （SUT） are proteins which are responsible for mediating sucrose transmembrane transport,and play a vital role in plant growth and developing.At present,5 members are identified in rice （Oryza sativa L.）,which are named as OsSUT 1～5,respectively,and the most functions of OsSUT3,OsSUT4 and OsSUT5 are not documented.In order to understand the role of OsSUT5 in rice callus induction and regeneration,experiments were conducted using antisense OsSUT5 transgenic rice and wild type rice MingHui86.The results showed that scutellum of antisense OsSUT5 transgenic rice became enlargement,and the induction and increase rate of callus was significantly declined（P＜0.05）.The dry weight of the first and second generation callus of transgenic rice were 0.028 80 and 0.119 93 g/peridish,respectively,significantly lower than that of the wild type（0.058 70 and 0.174 55 g/peridish）（P＜0.05）.After subculturing five generations,proliferation rate of callus flesh weight of transgenic rice was not significently different compared with wild type rice callus,however,the callus vitality of transgenic rice was significantly lower than that of wild type rice,and the callus of transgenic rice had obviously hygrophanous shape,and the green seedling differentiation rate was 65％.The callus differentiation rate was significantly decreased in the transgenic rice compared to the wild type（82.5％）（P＜0.05）.Real-time PCR analysis of the OsSUTs expression of the two generations of callus showed that the OsSUT5 expression decreased nearly one time of transgenic rice compared to the wild type,and OsSUT1 gene expression exhibited the same trend with the OsSUT5,while the other three OsSUTs genes were not influenced by OsSUT5 down expression.Our results suggested that sucrose transporter OsSUT5 can play an important role in regulating the rice callus induction and regeneration.This study provide the reference data for further analysis of OsSUT5 gene function.

关 键 词：水稻 愈伤组织 蔗糖转运蛋白基因(OsSUT5) 植株再生 

分 类 号：S567.239[农业科学—中草药栽培]