巴西橡胶树HbSERK2的克隆与表达分析
|
摘要
为了探究体细胞胚胎发生相关类受体蛋白激酶基因在橡胶树体细胞胚发生中的作用,在橡胶树体细胞胚转录组测序的基础上,克隆出一条编码巴西橡胶树体细胞胚胎发生相关类受体蛋白激酶(somatic embryogenesis receptor-like kinase, SERK)基因的序列,命名为HbSERK2。HbSERK2长1 404 bp,编码467个氨基酸,推测HbSERK2蛋白分子量为51.49 kD,等电点为5.53。Hb SERK2序列与木薯、毛果杨、胡杨、蓖麻、柑橘、麻风树等物种的体细胞胚胎发生相关类受体蛋白激酶具有较高的同源性,分别为99%、97%、97%、97%、97%、97%。生物信息学分析表明,Hb SERK2含有一个高度保守的蛋白激酶家族结构域,并且含有ATP结合区、蛋白激酶活化位点和Ser/Thr活性位点。q RT-PCR结果显示,HbSERK2在50 d愈伤、初级胚、球形胚、子叶形胚和幼苗中均有表达,在初级胚中表达量最高。本研究结果表明HbSERK2可能在橡胶树体细胞胚发生早期发挥重要作用。
To study the function of somatic embryogenesis receptor-like kinase gene in Hevea brasiliensis' s somatic embryogenesis, based on the transcriptome sequencing of Hevea brasiliensis somatic embryos, the full-length cDNA encoding a somatic embryogenesis receptor-like kinase(SERK), designated as HbSERK2, was isolated from Hevea brasiliensis. HbSERK2 contained a 1 404 bp open reading frame encoding 467 amino acids with molecular mass of 51.49 kD and pI was 5.53. The deduced amino acid sequences of HbSERK2 shared 99%, 97%, 97%, 97%,97% and 97% identity with the SERKs from Manihot esculenta, Populus trichocarpa, Populus euphratica, Ricinus communis, Citrus unshiu and Jatropha curcas. Hb SERK2 contained a conserved intracellular serine/threonine protein kinase domain. HbSERK2 was expressed at different levels, with the highest transcription in the primary embryo. It was suggested that HbSERK2 may play an important role in the early stage of somatic embryogenesis in Hevea brasiliensis.
To study the function of somatic embryogenesis receptor-like kinase gene in Hevea brasiliensis' s somatic embryogenesis, based on the transcriptome sequencing of Hevea brasiliensis somatic embryos, the full-length cDNA encoding a somatic embryogenesis receptor-like kinase(SERK), designated as HbSERK2, was isolated from Hevea brasiliensis. HbSERK2 contained a 1 404 bp open reading frame encoding 467 amino acids with molecular mass of 51.49 kD and pI was 5.53. The deduced amino acid sequences of HbSERK2 shared 99%, 97%, 97%, 97%,97% and 97% identity with the SERKs from Manihot esculenta, Populus trichocarpa, Populus euphratica, Ricinus communis, Citrus unshiu and Jatropha curcas. Hb SERK2 contained a conserved intracellular serine/threonine protein kinase domain. HbSERK2 was expressed at different levels, with the highest transcription in the primary embryo. It was suggested that HbSERK2 may play an important role in the early stage of somatic embryogenesis in Hevea brasiliensis.
引文
Baudino S.,Hansen S.,Brettschneider R.,Hecht V.F.G.,Dresselhaus T.,Lorz H.,Dumas C.,and Rogowsky P.M.,2001,Molecular characterisation of two novel maize LRR receptor-like kinases,which belong to the SERK gene family,Planta,213(1):1-10
Boutilier K.,Offringa R.,Sharma V.K.,Kieft H.,Ouellet T.,Zhang L.,Hattori J.,Liu C.M.,Van Lammeren A.A.M.,Miki B.L.A.,Custers J.B.M.,and Van Lookeren Campagne M.M.,2002,Ectopic expression of BABY BOOMtriggers a conversion from vegetative to embryonic growth,Plant Cell,14(8):1737-1749
Gao Y.,Xi M.L.,Wang G.F.,Yang L.W.,and Shi J.S.,2010,Molecular characterization and expression analysis of PmSERK1 during somatic embryogenesis in masson pine,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),8(1):53-58(高燕,席梦利,王桂凤,杨立伟,施季森,2010,马尾松体细胞胚胎发生相关基因PmSERK1的克隆与表达分析,分子植物育种,8(1):53-58)
Gu J.L.,2010,Studies on EC long-term subculture and proteomics during somatic embryogenesis in Litchi chinensis Sonn.,Thesis for M.S.,Fujian Agriculture and Forestry U-niversity,Supervisor:Lai Z.X.,pp.9-10(顾金玲,2010,荔枝EC长期继代保持与体胚发生及其蛋白质组学研究,硕士学位论文,福建农林大学,导师:赖钟雄,pp.9-10)
Hecht V.,Philippe J.,Calzada V.,Hartog M.V.,Schmidt E.D.L.,Boutilier K.,Grossniklaus U.,and De Vries S.C.,2001,The Arabidopsis SOMATIC EMBRYOGENESIS RECEP-TOR KINASE 1 gene is expressed in developing ovules and embryos and enhances embryogenic competence in culture,Plant Physiol.,127(3):803-816
Hu H.,Xiong L.,and Yang Y.,2005,Rice SERK1 gene positively regulates somatic embryogenesis of cultured cell and host defense response against fungal infection,Planta,222(1):107-117
Keith K.,Kraml M.,Dengler N.G.,and McCourt P.,1994,Fusca3:a hetetachmnic mutation affecting late embryo development in Arabidopsis,Plant Cell,6(5):589-600
Li L.,Zhang L.F.,Qi L.W.,and Han S.Y.,2013,Characterization and expression of the somatic embryogenesis receptor like kinase 1(SERK1)gene during somatic embryogenesis of Larix kaempferi,Linye Kexue Yanjiu(Forest Research),26(6):673-680(李龙,张立峰,齐力旺,韩素英,2013,日本落叶松体细胞胚胎发生相关基因LaSERK1的克隆与表达分析,林业科学研究,26(6):673-680)
Lin Q.G.,Cui B.M.,and Peng M.,2007,Advanced study on SERK genes family,Yichuan(Hereditas),29(6):681-687(林庆光,崔百明,彭明,2007,SERK基因家族的研究进展,遗传,29(6):681-687)
Lu M.,Xu C.Y.,and Guo W.D.,2006,Research progress of plant LRR receptor protein kinases,Zhejiang Shifan Daxue Xuebao(Journal of Zhejiang Normal University(Natural Sciences)),29(3):322-325(路梅,徐传雨,郭卫东,2006,植物LRR类受体蛋白激酶的研究进展,浙江师范大学学报(自然科学版),29(3):322-325)
Ma Y.Y.,Gan R.,and Wang N.N.,2005,Biological function of leucine-rich repeat like receptor like protein kinases in plants,Zhiwu Shengli Yu Fenzi Shengwuxue Xuebao(Journal of Plant Physiology and Molecular Biology),31(4):331-339(马媛媛,甘睿,王宁宁,2005,植物富含亮氨酸重复序列型类受体蛋白激酶的生物学功能,植物生理与分子生物学学报,31(4):331-339)
Philip B.W.,1998,Receptor kinases in plant development,Trends Plant Sci.,3(10):384-388
Santos M.D.O.,Romano E.,Yotoko K.S.C.,Tinoco M.L.P.,Dias B.B.A.,and Arag觔o F.J.,2005,Characterisation of the Cacao somatic embryogenesis receptor-like kinase(SERK)gene expressed during somatic embryogenesis,Plant Sci.,168(3):723-729
Schmidt E.D.,Guzzo F.,Toonen M.A.,and De Vries S.C.,1997,A leucine-rich repeat containing receptor like kinase marks somatic plant cells competent to form embryos,Development,124(10):2049-2062
Shimada T.,Hirabayashi T.,Endo T.,Fujii H.,Kita M.,and O-mura M.,2005,Isolation and characterization of the somatic embryogenesis receptor like kinase gene homologue(CitSERK1)from Citrus unshiu Marc.,Sci.Hortic.,103(2):233-238
Shiu S.H.,and Bleecker A.B.,2001,Receptor-like kinases from Arabidopsis form a monophyletic gene family related to animal receptor kinases,Proc.Natl.Acad.Sci.USA,98(19):10763-10768
Tian N.,Wang A.X.,Zhang K.Z.,Cui J.T.,and Jia Y.H.,2015,Clone and expression of AaSERK gene in Anthurium andraeanum's somatic embryogenesis,Xibei Zhiwu Xuebao(Acta Botanica Boreali-Occidentalia Sinica),35(4):674-681(田娜,王爱香,张克中,崔金腾,贾月慧,2015,火鹤AaSERK基因克隆及其在体细胞胚胎发生中的表达分析,西北植物学报,35(4):674-681)
Tichtinsky G.,Vanoosthuyse V.,Cock J.M.,and Gaude T.,2003,Making inroads into plant receptor kinase signalling pathways,Trends Plant Sci.,8(5):231-237
Wang X.J.,Zhang J.X.,He C.M.,Zeng S.J.,Wu K.L.,and Duan J.,2015,Cloning and expression analysis of somatic embryogenesis receptor kinase gene(DoSERK)from Dendrobium officinale Kimura et Migo,Redai Yaredai Zhiwu Xuebao(Journal of Tropical and Subtropical Botany),23(5):518-526(王晓娟,张建霞,何春梅,曾宋君,吴坤林,段俊,2015,铁皮石斛体细胞胚胎发生类受体激酶基因DoSERK的克隆和表达分析,热带亚热带植物学报,23(5):518-526)
Wu C.T.,Li W.G.,and Huang H.S.,2012,New advance on Hevea brasiliensis of recent years at home and abroad,Guangxi Zhiwu(Guihaia),32(6):860-866(吴春太,李维国,黄华孙,2012,近年来国内外橡胶树育种研究新进展,广西植物,32(6):860-866)
Yang Y.W.,He B.,Zhang B.L.,and Ni W.C.,2010,Molecular cloning and expression analysis of a receptor like protein kinase gene in upland cotton,Mianhua Xuebao(Cotton Science),23(1):15-21(杨郁文,何冰,张保龙,倪万潮,2010,一个棉花类受体蛋白激酶基因的克隆与表达分析,棉花学报,23(1):15-21)
Zha X.J.,Ma B.J.,Pan J.W.,and Yang J.S.,2010,Research progress of plant leucine rich repeat receptor protein kinases,Zhejiang Shifan Daxue Xuebao(Journal of Zhejiang Normal University(Natural Sciences)),33(1):7-12(查笑君,马伯军,潘建伟,杨金水,2010,植物富亮氨酸重复类受体蛋白激酶的研究进展,浙江师范大学学报(自然科学版),33(1):7-12)
Zuo J.R.,Niu Q.W.,Frugis G.,and Chua N.H.,2002,The WUSCHEL gene promotes vegetative-to-embryonic transition in Arabidopsis,Plant J.,30(3):349-359
Boutilier K.,Offringa R.,Sharma V.K.,Kieft H.,Ouellet T.,Zhang L.,Hattori J.,Liu C.M.,Van Lammeren A.A.M.,Miki B.L.A.,Custers J.B.M.,and Van Lookeren Campagne M.M.,2002,Ectopic expression of BABY BOOMtriggers a conversion from vegetative to embryonic growth,Plant Cell,14(8):1737-1749
Gao Y.,Xi M.L.,Wang G.F.,Yang L.W.,and Shi J.S.,2010,Molecular characterization and expression analysis of PmSERK1 during somatic embryogenesis in masson pine,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),8(1):53-58(高燕,席梦利,王桂凤,杨立伟,施季森,2010,马尾松体细胞胚胎发生相关基因PmSERK1的克隆与表达分析,分子植物育种,8(1):53-58)
Gu J.L.,2010,Studies on EC long-term subculture and proteomics during somatic embryogenesis in Litchi chinensis Sonn.,Thesis for M.S.,Fujian Agriculture and Forestry U-niversity,Supervisor:Lai Z.X.,pp.9-10(顾金玲,2010,荔枝EC长期继代保持与体胚发生及其蛋白质组学研究,硕士学位论文,福建农林大学,导师:赖钟雄,pp.9-10)
Hecht V.,Philippe J.,Calzada V.,Hartog M.V.,Schmidt E.D.L.,Boutilier K.,Grossniklaus U.,and De Vries S.C.,2001,The Arabidopsis SOMATIC EMBRYOGENESIS RECEP-TOR KINASE 1 gene is expressed in developing ovules and embryos and enhances embryogenic competence in culture,Plant Physiol.,127(3):803-816
Hu H.,Xiong L.,and Yang Y.,2005,Rice SERK1 gene positively regulates somatic embryogenesis of cultured cell and host defense response against fungal infection,Planta,222(1):107-117
Keith K.,Kraml M.,Dengler N.G.,and McCourt P.,1994,Fusca3:a hetetachmnic mutation affecting late embryo development in Arabidopsis,Plant Cell,6(5):589-600
Li L.,Zhang L.F.,Qi L.W.,and Han S.Y.,2013,Characterization and expression of the somatic embryogenesis receptor like kinase 1(SERK1)gene during somatic embryogenesis of Larix kaempferi,Linye Kexue Yanjiu(Forest Research),26(6):673-680(李龙,张立峰,齐力旺,韩素英,2013,日本落叶松体细胞胚胎发生相关基因LaSERK1的克隆与表达分析,林业科学研究,26(6):673-680)
Lin Q.G.,Cui B.M.,and Peng M.,2007,Advanced study on SERK genes family,Yichuan(Hereditas),29(6):681-687(林庆光,崔百明,彭明,2007,SERK基因家族的研究进展,遗传,29(6):681-687)
Lu M.,Xu C.Y.,and Guo W.D.,2006,Research progress of plant LRR receptor protein kinases,Zhejiang Shifan Daxue Xuebao(Journal of Zhejiang Normal University(Natural Sciences)),29(3):322-325(路梅,徐传雨,郭卫东,2006,植物LRR类受体蛋白激酶的研究进展,浙江师范大学学报(自然科学版),29(3):322-325)
Ma Y.Y.,Gan R.,and Wang N.N.,2005,Biological function of leucine-rich repeat like receptor like protein kinases in plants,Zhiwu Shengli Yu Fenzi Shengwuxue Xuebao(Journal of Plant Physiology and Molecular Biology),31(4):331-339(马媛媛,甘睿,王宁宁,2005,植物富含亮氨酸重复序列型类受体蛋白激酶的生物学功能,植物生理与分子生物学学报,31(4):331-339)
Philip B.W.,1998,Receptor kinases in plant development,Trends Plant Sci.,3(10):384-388
Santos M.D.O.,Romano E.,Yotoko K.S.C.,Tinoco M.L.P.,Dias B.B.A.,and Arag觔o F.J.,2005,Characterisation of the Cacao somatic embryogenesis receptor-like kinase(SERK)gene expressed during somatic embryogenesis,Plant Sci.,168(3):723-729
Schmidt E.D.,Guzzo F.,Toonen M.A.,and De Vries S.C.,1997,A leucine-rich repeat containing receptor like kinase marks somatic plant cells competent to form embryos,Development,124(10):2049-2062
Shimada T.,Hirabayashi T.,Endo T.,Fujii H.,Kita M.,and O-mura M.,2005,Isolation and characterization of the somatic embryogenesis receptor like kinase gene homologue(CitSERK1)from Citrus unshiu Marc.,Sci.Hortic.,103(2):233-238
Shiu S.H.,and Bleecker A.B.,2001,Receptor-like kinases from Arabidopsis form a monophyletic gene family related to animal receptor kinases,Proc.Natl.Acad.Sci.USA,98(19):10763-10768
Tian N.,Wang A.X.,Zhang K.Z.,Cui J.T.,and Jia Y.H.,2015,Clone and expression of AaSERK gene in Anthurium andraeanum's somatic embryogenesis,Xibei Zhiwu Xuebao(Acta Botanica Boreali-Occidentalia Sinica),35(4):674-681(田娜,王爱香,张克中,崔金腾,贾月慧,2015,火鹤AaSERK基因克隆及其在体细胞胚胎发生中的表达分析,西北植物学报,35(4):674-681)
Tichtinsky G.,Vanoosthuyse V.,Cock J.M.,and Gaude T.,2003,Making inroads into plant receptor kinase signalling pathways,Trends Plant Sci.,8(5):231-237
Wang X.J.,Zhang J.X.,He C.M.,Zeng S.J.,Wu K.L.,and Duan J.,2015,Cloning and expression analysis of somatic embryogenesis receptor kinase gene(DoSERK)from Dendrobium officinale Kimura et Migo,Redai Yaredai Zhiwu Xuebao(Journal of Tropical and Subtropical Botany),23(5):518-526(王晓娟,张建霞,何春梅,曾宋君,吴坤林,段俊,2015,铁皮石斛体细胞胚胎发生类受体激酶基因DoSERK的克隆和表达分析,热带亚热带植物学报,23(5):518-526)
Wu C.T.,Li W.G.,and Huang H.S.,2012,New advance on Hevea brasiliensis of recent years at home and abroad,Guangxi Zhiwu(Guihaia),32(6):860-866(吴春太,李维国,黄华孙,2012,近年来国内外橡胶树育种研究新进展,广西植物,32(6):860-866)
Yang Y.W.,He B.,Zhang B.L.,and Ni W.C.,2010,Molecular cloning and expression analysis of a receptor like protein kinase gene in upland cotton,Mianhua Xuebao(Cotton Science),23(1):15-21(杨郁文,何冰,张保龙,倪万潮,2010,一个棉花类受体蛋白激酶基因的克隆与表达分析,棉花学报,23(1):15-21)
Zha X.J.,Ma B.J.,Pan J.W.,and Yang J.S.,2010,Research progress of plant leucine rich repeat receptor protein kinases,Zhejiang Shifan Daxue Xuebao(Journal of Zhejiang Normal University(Natural Sciences)),33(1):7-12(查笑君,马伯军,潘建伟,杨金水,2010,植物富亮氨酸重复类受体蛋白激酶的研究进展,浙江师范大学学报(自然科学版),33(1):7-12)
Zuo J.R.,Niu Q.W.,Frugis G.,and Chua N.H.,2002,The WUSCHEL gene promotes vegetative-to-embryonic transition in Arabidopsis,Plant J.,30(3):349-359