水稻锌指蛋白基因OsSZP载体构建、遗传转化及功能分析
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摘要
文章通过RNA干扰(RNA interference,RNAi)技术对水稻锌指蛋白基因OsSZP的功能进行分析。定量聚合链式反应(polymerase chain reaction,PCR)分析结果表明,通过RNAi技术可获得OsSZP表达水平明显降低的转基因株系。与野生型相比,OsSZP基因下调植株的秕谷率显著增加,花粉育性降低,表明锌指蛋白基因OsSZP对水稻雄配子发育具有调控作用。表达模式分析结果表明,OsSZP在水稻幼苗和成熟植株根中均不表达,在水稻花药中表达水平较高。此外,OsSZP基因的表达也受温度、光照的影响。
The function of a rice zinc-finger protein gene OsSZP was investigated by RNA interference(RNAi).The analysis results of quantitative polymerase chain reaction(PCR)and phenotype showed that the transgenic plants with significantly lower OsSZPexpression level could be obtained by RNAi.The OsSZPdown-regulated transgenic plants exhibited increased blasted rates and reduced pollen fertilities compared with wildtype,suggesting that OsSZPfunctions as a regulator of male gamete development in rice.The expression pattern analysis results revealed that OsSZP was not expressed in roots of rice seedlings and mature plants,but showed higher level in rice anther.In addition,the abundance of OsSZP was also affected by temperature and light.
The function of a rice zinc-finger protein gene OsSZP was investigated by RNA interference(RNAi).The analysis results of quantitative polymerase chain reaction(PCR)and phenotype showed that the transgenic plants with significantly lower OsSZPexpression level could be obtained by RNAi.The OsSZPdown-regulated transgenic plants exhibited increased blasted rates and reduced pollen fertilities compared with wildtype,suggesting that OsSZPfunctions as a regulator of male gamete development in rice.The expression pattern analysis results revealed that OsSZP was not expressed in roots of rice seedlings and mature plants,but showed higher level in rice anther.In addition,the abundance of OsSZP was also affected by temperature and light.
引文
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[24]LEVY Y Y,MESNAGE S,MYLNW J S,et al.Multiple Roles of Arabidopsis VRN1in vernalization and flowering time control[J].Science,2002,297(5579):243-246.
[2]TAKATSUJI H.Zinc-finger proteins:the classical zinc finger emerges in contemporary plant science[J].Plant Molecular Biology,1999,39(6):1073-1078.
[3]LIU Liansen,WHITE M J,MACRAE T H.Transcription factors and their genes in higher plants:functional domains,evolution and regulation[J].European Journal of Biochemistry,1999,262(2):247-257.
[4]GAMSIAEGER R,LIEW C K,LOUGHLIN F E,et al.Sticky fingers:zinc-fingers as protein-recognition motifs[J].Trend in Biochemical Sciences,2007,32(2):63-70.
[5]ENGLBRECHT C C,SCHOOF H,BOHM S.Conservation,diversification and expansion of C2H2zinc finger proteins in the Arabidopsis thaliana genome[J].BMC Genomics,2004,5:39.
[6]Agarwal P,Arora R,Ray S,et al.Genome-wide identification of C2H2zinc-finger gene family in rice and their phylogeny and expression analysis[J].Plant Molecular Biology,2007,65:467-485.
[7]WANG D,GUO Y,WU C,et al.Genome-wide analysis of CCCH zinc finger family in Arabidopsis and rice[J].BMC Genomics,2008,9:44-54.
[8]CIFTCI-YILMAZ S,MITTLER R.The zinc finger network of plants[J].Cellular and Molecular Life Sciences,2008,65:1150-1160.
[9]TAKATSUJI H.Zinc-finger transcription factors in plant[J].Cellular and Molecular Life Sciences,1998,54(6):582-596.
[10]SINGH K B,FOLEY R C,O珘nate-sanchez L.Transcription factors in plant defense and stress responses[J].Current Opinion in Plant Biology,2002,5(5):430-436.
[11]YANO M,KATAYOSE Y,ASHIKARI M,et al.Hd1,a major photoperiod sensitivity quantitative trait locus in rice,is closely related to the Arabidopsis flowering time gene CONSTANS[J].The Plant Cell,2000,12(12):2473-2484.
[12]KOZAKI A,HAKE S,COLASANTI J.The maize ID1flowering time regulator is a zinc finfer protein with novel DNA binding properties[J].Nucleic Acids Research,2004,32(5):1710-1720.
[13]WU C,YOU C,LI C,et al.RID1,encoding a Cys Z-His2-type zinc finger transcription factor,acts as a master switch from vegetative to floral development in rice[J].Proceedings of the National Academy of Sciences of the United States of America,2008,105(35):12915-12920.
[14]KAPOOR S,KOBAYASHI A,TAKATUSJI H.Silencing of the tapetum-specific zinc finger gene TAZ1causes premature degeneration of tapetum and pollen abortion in petunia[J].The Plant Cell,2002,14(10):2353-2367.
[15]ITO T,NAGATA N,YOSHIBA Y,et al.Arabidopsis MALE STERILITY1encodes a PHD-type transcription factor and regulates pollen and tapetum development[J].The Plant Cell,2007,19(11):3549-3562.
[16]胡骏,黄文超,朱仁山,等.水稻雄性不育与杂种优势的利用[J].武汉大学学报(理学版),2013,59(1):1-9.
[17]FIRE A,XU S Q,MONTGOMERY M K,et al.Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans[J].Nature,1998,391:806-811.
[18]MAO J,ZHANG Y C,SANG Y,et al.A role for Arabidopsis cryptochromes and COP1in the regulation of stomatal opening[J].Proceedings of the National Academy of Sciences of the United States of America,2005,102(34):12270-12275.
[19]HIEI Y,OHTA S,KOMARI T,et al.Efficient transformation of rice(Oryza sativa L.)mediated by Agrobacterium and sequence analysis of the boundaries of the TDNA[J].The Plant Journal,1994,6(2):271-282.
[20]张惠莹,张黎,李嵘,等.西南地区4个籼稻恢复系遗传转化植株再生体系研究[J].合肥工业大学学报(自然科学版),2013,36(12):1511-1517.
[21]谭学林.滇型杂交水稻雄性不育系子房孤雌发育的观察[J].云南农业大学学报,1989,4(1):82-83.
[22]黄方,迟英俊,喻德跃.植物MADS-box基因研究进展[J].南京农业大学学报,2012,35(5):9-18.
[23]MAES T,ZETHOF J,KARIMI M,et al.Petunia Ap2-like genes and their role in flower and seed development[J].The Plant Cell,2001,13(2):229-244.
[24]LEVY Y Y,MESNAGE S,MYLNW J S,et al.Multiple Roles of Arabidopsis VRN1in vernalization and flowering time control[J].Science,2002,297(5579):243-246.