JAZ基因家族6个成员在橡胶树上的物理定位
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摘要
茉莉酸信号途径与橡胶树的防御能力及胶乳合成密切相关,JAZ蛋白是茉莉酸信号途径下游基因转录调控关键的抑制因子,目前虽已报道多个JAZ基因,但该家族基因在染色体上的具体位置尚未明确。本研究中,以巴西橡胶树的"热研7-33-97"品种作为实验材料,分别利用原位PCR技术和和荧光原位杂交技术对巴西橡胶树的JAZ家族的6个基因(jaz1,jaz2,jaz3,jaz5,jaz6,jaz7)在染色体上的位置进行了物理定位分析,结果表明:HbJAZ1、HbJAZ2、HbJAZ3、HbJAZ5、HbJAZ6和HbJAZ7基因分别位于巴西橡胶树(热研7-33-97)的第4号染色体的长臂、第4号染色体的长臂、第8号染色体的长臂、第9号染色体的短臂、第6号染色体的长臂上和第1号染色体的短臂上;信号位点距离着丝点的平均百分比距分别为:47.29、41.87、51.98、81.74、20.07和29.11。基因与其它基因间的连锁关系,并有效的应用于橡胶分子遗传育种方面的研究。
The jasmonate pathway is closely related to the defense function and the production of plastic function of rubber tree, and JAZ protein is the key inhibitor of the transcriptional regulation of downstream genes. Although several JAZ genes have been reported, the specific location of the family gene on the chromosome is not yet clear.In this study, we took the "Reyan 7-33-97"of Hevea brasiliensis as experimental materials, the location of six genes(jaz1, jaz2, jaz3, jaz5, jaz6 and jaz7) of the JAZ family in the chromosome were analysed by using in situ PCR technology and fluorescence in situ hybridization technology. The results showed that HbJAZ1, HbJAZ2, HbJAZ3,HbJAZ5, HbJAZ6 and HbJAZ7 genes were located separately in the long arm of chromosome 4, long arm of chromosome 4, long arm of chromosome 8, short arm of chromosome 9, long arm of chromosome 6 and the short arm of chromosome 1 of the Hevea brasiliensis; the average percentage distance of the signal loci from the centromere were 47.29, 41.87, 51.98, 81.74, 20.07, 29.11, respectively. the linkage between these genes and other genes, which can effectively applied to the research of rubber molecular genetic breeding.
The jasmonate pathway is closely related to the defense function and the production of plastic function of rubber tree, and JAZ protein is the key inhibitor of the transcriptional regulation of downstream genes. Although several JAZ genes have been reported, the specific location of the family gene on the chromosome is not yet clear.In this study, we took the "Reyan 7-33-97"of Hevea brasiliensis as experimental materials, the location of six genes(jaz1, jaz2, jaz3, jaz5, jaz6 and jaz7) of the JAZ family in the chromosome were analysed by using in situ PCR technology and fluorescence in situ hybridization technology. The results showed that HbJAZ1, HbJAZ2, HbJAZ3,HbJAZ5, HbJAZ6 and HbJAZ7 genes were located separately in the long arm of chromosome 4, long arm of chromosome 4, long arm of chromosome 8, short arm of chromosome 9, long arm of chromosome 6 and the short arm of chromosome 1 of the Hevea brasiliensis; the average percentage distance of the signal loci from the centromere were 47.29, 41.87, 51.98, 81.74, 20.07, 29.11, respectively. the linkage between these genes and other genes, which can effectively applied to the research of rubber molecular genetic breeding.
引文
Ga ll J.G.,and Pardue M.L.,1969,Formation and detection of RAN-DAN hybrid molecules in cytological preparations,Proc.Natl.Acad.Sci.USA,63(2):378-383
Gao H.Q.,Wang Y.,Jin G.,Qiu H.Y.,and Zhuang N.S.,2009a,Optimization of chromosome producing method for rubber tree leaves,Redai Zuowu Xuebao(Journal of Tropical Crops),30(5):565-569(高和琼,王英,金鸽,邱海燕,庄南生,2009a,橡胶树叶片染色体制片方法的优化,热带作物学报,30(5):565-569)
Gao H.Q.,Zhuang N.S.,and Wang Y.,2009b,Karyotype analysis of two varieties of rubber tree,Wuhan Zhiwuxue Yanjiu(Wuhan Botanical Research),27(5):537-540(高和琼,庄南生,王英,2009b,橡胶树两个品种的核型分析,武汉植物学研究,27(5):537-540)
Gao J.J.,2014,Study on the Physical positioning of several rubber-related genes in Hevea brasiliensis,Thesis for M.S.,Hainan University,Supervisor:Zhuang N.S.,pp.24-31(高佳佳,2014,巴西橡胶树几个与橡胶合成相关基因的物理定位研究,硕士学位论文,海南大学,导师:庄南生,pp.24-31)
Gao J.J.,Wang Y.,Gao H.Q.,Zhu W.,Zhuang N.S.,2014,Study on the physical location of the HbNIN gene family of rubber latex in Brazilian rubber tree,Redai Zuowu Xuebao(Journal of Tropical Crops),35(9):1704-1709(高佳佳,王英,高和琼,朱稳,庄南生,2014,巴西橡胶树胶乳转化酶HbNIN基因家族物理定位的研究,热带作物学报,35(9):1704-1709)
Greelman R.A.,and Mullet J.E.,1997,Biosynthesis and action jasmonates in plants,Annu.Rev.Plant Physiol.Plant Mol.Biol.,48:355-381
Guan J.Y.,Wang Y.,Gao H.Q.,and Zhuang N.S.,2014,Physical localization of four rings of zinc finger protein gene(HbRZF)in Brazilian rubber tree,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),33(3):610-616(官锦燕,王英,高和琼,庄南生,2014,巴西橡胶树4个环锌指蛋白基因(HbRZF)的物理定位,基因组学与应用生物学,33(3):610-616)
Hong H.,Xiao H.,Yuan H.,Zhai J.,and Huang X.,2014,Cloning and characterisation of JAZ gene family in Hevea brasiliensis,Plant Biol.,17(3):618-624
He S.B.,Chai L.Q.,Tan J.J.,Shen R.,Zhou P.,Ma Y.F.,and Li L.J.,2014,Advances in fluorescence in situ hybridization,Zhiwu Kexue Xuebao(Journal of Plant Science),32(2):199-204(何世斌,柴连琴,谭珺隽,沈尧,周萍,马云峰,李立家,2014,荧光原位杂交技术的研究进展,植物科学学报,32(2):199-204)
Li F.L.,and Chen Q.F.,2010,Application of in situ PCR in botany,Guizhou Shifan Daxue Xuebao(Journal of Guizhou Normal University(Natural Science Edition)),28(2):111-114(李分龙,陈庆富,2010,原位PCR技术在植物学中的应用,贵州师范大学学报(自然科学版),28(2):111-114)
Lin X.Q.,Mao J.,Lu X.,Liu X.L.,Ma L.,and Cai Q.,2012,Applications and prospects of fluorescence in situ hybridization on plant molecular cytogenetics,Ganzhe Tangye(Sugarcane and Canesugar),(1):51-57(林秀琴,毛钧,陆鑫,刘新龙,马丽,蔡青,2012,荧光原位杂交技术(FISH)及其在植物分子细胞遗传学中的应用与展望,甘蔗糖业,(1):51-57)
Ning H.,Ren N.,Song Y.C.,and Li L.J.,1996,The physical location of the maize nac1 genes by in situ hybridization,Huazhong Shifan Daxue Xuebao(Journal of Huazhong Normal University(Natural Science Edition)),30(4):480-485(宁华,任南,宋运淳,李立家,1996,玉米(Zea may L.)盐逆境胁迫蛋白基因nac1的染色体原位杂交物理定位,华中师范大学学报(自然科学版),30(4):480-485)
Peng B.F.,Wang Y.,Gao H.Q.,and Zhuang N.S.,2016,Physical locations of four members of hevein gene family in Hevea brasiliensis,Redai Shengwu Xuebao(Journal of Tropical Biology),7(3):318-324(彭宝丰,王英,高和琼,庄南生,2016,橡胶素基因家族4个成员在橡胶树染色体上的定位,热带生物学报,7(3):318-324)
Qiu H.Y.,2010,Study on the physical mapping of REF,RT and SRPP genes in brazilian rubber tree,Thesis for M.S.,Hainan University,Supervisor:Zhuang N.S.,pp.24-26(邱海燕,2010,巴西橡胶树REF、RT和SRPP基因物理定位的研究,硕士学位论文,海南大学,导师:庄南生,pp.24-26)
Song Y.C.,and Gustafson J.P.,1995,The physical location of fourteen RFLP markers in rice(Oryza sativa L.),Theor.Appl.Genet.,90(1):113-119
Wu W.Q.,Huang D.Y.,and Zhuang N.S.,2006,In situ PCR and its role in plant research,Redai Nongye Kexue(Tropical Agriculture Sciences),26(2):65-69(吴文蔷,黄东益,庄南生,2006,原位PCR及其在植物研究中的作用,热带农业科学,26(2):65-69)
Yang S.G.,Yu J.H.,Shi M.J.,Chen Y.Y.,and Tian W.M.,2008,Effects of dicing,mechanical injury and exogenous jasmonic acid on the expression of defensive protein gene in the cell of rubber tubal,Redai Zuowu Xuebao(Chinese Journal of Tropical Crops),29(5):535-540(杨署光,于俊红,史敏晶,陈月异,田维敏,2008,割胶、机械伤害和外源茉莉酸对橡胶树乳管细胞防卫蛋白基因表达的影响,热带作物学报,29(5):535-540)
Yao D.,2016,Cloning and mutation of HbJAZs gene in brazilian rubber tree,Thesis for M.S.,Hainan University,Supervisor:Huang X.,pp.5-7(姚笛,2016,巴西橡胶树HbJAZs基因的克隆与互作蛋白鉴定,硕士学位论文,海南大学,导师:黄惜,pp.5-7)
Yao D.,Wang Q.C.,Chao Y.X.,and Huang X.,2016,Cloning and expression analysis of JAZ family gene in Brazilian Rubber tree,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),14(5):1115-1122(姚笛,王启超,曹玉鑫,洪灏,黄惜,2016,巴西橡胶树JAZ家族基因的克隆和表达分析,分子植物育种,14(5):1115-1122)
Zhang X.Y.,Jiang H.B.,Li L.J.,Ma Q.,Yang J.,and Liu T.,1996,In situ amplification of DNA fragments specific for human Y chromosome in cellular nuclei by PCR,Sci.China C Life Sci.,39(1):45-52
Zhang X.X.,2013,Physical localization of four latex biosynthetic genes in Brazilian rubber tree,Thesis for M.S.,Hainan University,Supervisor:Zhuang N.S.,pp.47(张新新,2013,巴西橡胶树4个胶乳生物合成相关基因的物理定位,硕士学位论文,海南大学,导师:庄南生,pp.47)
Gao H.Q.,Wang Y.,Jin G.,Qiu H.Y.,and Zhuang N.S.,2009a,Optimization of chromosome producing method for rubber tree leaves,Redai Zuowu Xuebao(Journal of Tropical Crops),30(5):565-569(高和琼,王英,金鸽,邱海燕,庄南生,2009a,橡胶树叶片染色体制片方法的优化,热带作物学报,30(5):565-569)
Gao H.Q.,Zhuang N.S.,and Wang Y.,2009b,Karyotype analysis of two varieties of rubber tree,Wuhan Zhiwuxue Yanjiu(Wuhan Botanical Research),27(5):537-540(高和琼,庄南生,王英,2009b,橡胶树两个品种的核型分析,武汉植物学研究,27(5):537-540)
Gao J.J.,2014,Study on the Physical positioning of several rubber-related genes in Hevea brasiliensis,Thesis for M.S.,Hainan University,Supervisor:Zhuang N.S.,pp.24-31(高佳佳,2014,巴西橡胶树几个与橡胶合成相关基因的物理定位研究,硕士学位论文,海南大学,导师:庄南生,pp.24-31)
Gao J.J.,Wang Y.,Gao H.Q.,Zhu W.,Zhuang N.S.,2014,Study on the physical location of the HbNIN gene family of rubber latex in Brazilian rubber tree,Redai Zuowu Xuebao(Journal of Tropical Crops),35(9):1704-1709(高佳佳,王英,高和琼,朱稳,庄南生,2014,巴西橡胶树胶乳转化酶HbNIN基因家族物理定位的研究,热带作物学报,35(9):1704-1709)
Greelman R.A.,and Mullet J.E.,1997,Biosynthesis and action jasmonates in plants,Annu.Rev.Plant Physiol.Plant Mol.Biol.,48:355-381
Guan J.Y.,Wang Y.,Gao H.Q.,and Zhuang N.S.,2014,Physical localization of four rings of zinc finger protein gene(HbRZF)in Brazilian rubber tree,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),33(3):610-616(官锦燕,王英,高和琼,庄南生,2014,巴西橡胶树4个环锌指蛋白基因(HbRZF)的物理定位,基因组学与应用生物学,33(3):610-616)
Hong H.,Xiao H.,Yuan H.,Zhai J.,and Huang X.,2014,Cloning and characterisation of JAZ gene family in Hevea brasiliensis,Plant Biol.,17(3):618-624
He S.B.,Chai L.Q.,Tan J.J.,Shen R.,Zhou P.,Ma Y.F.,and Li L.J.,2014,Advances in fluorescence in situ hybridization,Zhiwu Kexue Xuebao(Journal of Plant Science),32(2):199-204(何世斌,柴连琴,谭珺隽,沈尧,周萍,马云峰,李立家,2014,荧光原位杂交技术的研究进展,植物科学学报,32(2):199-204)
Li F.L.,and Chen Q.F.,2010,Application of in situ PCR in botany,Guizhou Shifan Daxue Xuebao(Journal of Guizhou Normal University(Natural Science Edition)),28(2):111-114(李分龙,陈庆富,2010,原位PCR技术在植物学中的应用,贵州师范大学学报(自然科学版),28(2):111-114)
Lin X.Q.,Mao J.,Lu X.,Liu X.L.,Ma L.,and Cai Q.,2012,Applications and prospects of fluorescence in situ hybridization on plant molecular cytogenetics,Ganzhe Tangye(Sugarcane and Canesugar),(1):51-57(林秀琴,毛钧,陆鑫,刘新龙,马丽,蔡青,2012,荧光原位杂交技术(FISH)及其在植物分子细胞遗传学中的应用与展望,甘蔗糖业,(1):51-57)
Ning H.,Ren N.,Song Y.C.,and Li L.J.,1996,The physical location of the maize nac1 genes by in situ hybridization,Huazhong Shifan Daxue Xuebao(Journal of Huazhong Normal University(Natural Science Edition)),30(4):480-485(宁华,任南,宋运淳,李立家,1996,玉米(Zea may L.)盐逆境胁迫蛋白基因nac1的染色体原位杂交物理定位,华中师范大学学报(自然科学版),30(4):480-485)
Peng B.F.,Wang Y.,Gao H.Q.,and Zhuang N.S.,2016,Physical locations of four members of hevein gene family in Hevea brasiliensis,Redai Shengwu Xuebao(Journal of Tropical Biology),7(3):318-324(彭宝丰,王英,高和琼,庄南生,2016,橡胶素基因家族4个成员在橡胶树染色体上的定位,热带生物学报,7(3):318-324)
Qiu H.Y.,2010,Study on the physical mapping of REF,RT and SRPP genes in brazilian rubber tree,Thesis for M.S.,Hainan University,Supervisor:Zhuang N.S.,pp.24-26(邱海燕,2010,巴西橡胶树REF、RT和SRPP基因物理定位的研究,硕士学位论文,海南大学,导师:庄南生,pp.24-26)
Song Y.C.,and Gustafson J.P.,1995,The physical location of fourteen RFLP markers in rice(Oryza sativa L.),Theor.Appl.Genet.,90(1):113-119
Wu W.Q.,Huang D.Y.,and Zhuang N.S.,2006,In situ PCR and its role in plant research,Redai Nongye Kexue(Tropical Agriculture Sciences),26(2):65-69(吴文蔷,黄东益,庄南生,2006,原位PCR及其在植物研究中的作用,热带农业科学,26(2):65-69)
Yang S.G.,Yu J.H.,Shi M.J.,Chen Y.Y.,and Tian W.M.,2008,Effects of dicing,mechanical injury and exogenous jasmonic acid on the expression of defensive protein gene in the cell of rubber tubal,Redai Zuowu Xuebao(Chinese Journal of Tropical Crops),29(5):535-540(杨署光,于俊红,史敏晶,陈月异,田维敏,2008,割胶、机械伤害和外源茉莉酸对橡胶树乳管细胞防卫蛋白基因表达的影响,热带作物学报,29(5):535-540)
Yao D.,2016,Cloning and mutation of HbJAZs gene in brazilian rubber tree,Thesis for M.S.,Hainan University,Supervisor:Huang X.,pp.5-7(姚笛,2016,巴西橡胶树HbJAZs基因的克隆与互作蛋白鉴定,硕士学位论文,海南大学,导师:黄惜,pp.5-7)
Yao D.,Wang Q.C.,Chao Y.X.,and Huang X.,2016,Cloning and expression analysis of JAZ family gene in Brazilian Rubber tree,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),14(5):1115-1122(姚笛,王启超,曹玉鑫,洪灏,黄惜,2016,巴西橡胶树JAZ家族基因的克隆和表达分析,分子植物育种,14(5):1115-1122)
Zhang X.Y.,Jiang H.B.,Li L.J.,Ma Q.,Yang J.,and Liu T.,1996,In situ amplification of DNA fragments specific for human Y chromosome in cellular nuclei by PCR,Sci.China C Life Sci.,39(1):45-52
Zhang X.X.,2013,Physical localization of four latex biosynthetic genes in Brazilian rubber tree,Thesis for M.S.,Hainan University,Supervisor:Zhuang N.S.,pp.47(张新新,2013,巴西橡胶树4个胶乳生物合成相关基因的物理定位,硕士学位论文,海南大学,导师:庄南生,pp.47)