cDNA Cloning and Expression Analysis of CjAPL1 Gene from Camellia japonica

SUN Ying-kun; LI Ji-yuan; YIN Heng-fu; FAN Zheng-qi; ZHOU Xing-wen

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cDNA Cloning and Expression Analysis of CjAPL1 Gene from Camellia japonica

1.
Research Institute of Subtropical Forestry, Chinese Academy of Forestry, Fuyang 311400, Zhejiang, China
2.
College of Landscape Architecture and Forestry, Qingdao Agricultural University, Qingdao 266109, Shandong, China

Received Date: 2012-11-01

Abstract

In order to study the role of A-class genes in the process of forming plena flower for Camellia japonica, a full-length cDNA sequence of APETALA1 (AP1) gene was cloned from early flower bud of C. japonica ‘Jinpan Lizhi’ using RT-PCR and RACE,named CjAPL1 (GenBank accession # JX657332). The results of nucleotide sequence analysis show that the length of CjAPL1 is 1 149 bp, containing an Opening Reading Frame (ORF) of 741 bp, a 5'-Untranslated Region (5'-UTR) of 206 bp, and a 3'-UTR of 202 bp. The CjAPL1 gene encodes a protein of 246 amino acids, and has more than 75% homology with A-class genes from Actinidia chinensis and Hydrangea macrophylla. Quantitative real-time PCR analysis shows that the gene expression pattern is that in early flower bud of III stage> early flower bud of II stage > early flower bud of I stage > squaring stage, and the tendency shows increasing slowly at the early stage and then decreasing sharply. In different floral organs, the highest expression was observed in style, next in ovary and sepals, and the lowest in bottom stamens and upper outer flowers. The difference of gene expression pattern indicates that CjAPL1 may play a role in the morphogenesis of pleiopetalous flower of Camellia.
- Camellia japonica,
- CjAPL1 gene,
- sequence analysis,
- relative real-time PCR,
- expression

References

[1]	Hu J Y, Saedler H. Evolution of the inflated-calyx-syndrome (ICS) in Solanaceae[J]. Mol Biol Evol, 2007, 24(11):2443-2453
[2]	Viaene T, Vekemans D, Irish V F, et al. Pistillata-duplications as a mode for floral diversification in (Basal) asterids[J]. Mol Biol Evol, 2009, 26(11):2627-2645
[3]	Coen E S, Meyerowitz E M. The war of the whorls: Genetic interactions controlling flower development[J]. Nature, 1991, 353(6339):31-37
[4]	Pelaz S, Ditta G S, Baumann E, et al. B and C floral organ identity functions require SEPALLATA MADS-box genes[J].Nature, 2000, 405(6783):200-203
[5]	Theissen G, Saedler H. Floral quartets[J]. Nature, 2001, 409(6819):469-471
[6]	Ditta G, Pinyopich A, Robles P, et al. The SEP4 gene of Arabidopsis thaliana functions in floral organ and meristem identity[J]. Curr Biol, 2004,14(21):1935-1940
[7]	Jack T. Molecular and genetic mechanisms of floral control[J]. Plant Cell, 2004, 16(Suppl):1-17
[8]	徐勇. 桃花发育相关MADS box基因研究 [D].北京:首都师范大学, 2007:23
[9]	Colombo L, Franken J, Koeteje E, et al. The petunia MADS box gene FBP11 determines ovule identity[J]. Plant Cell,1995, 7(11):1859-1868
[10]	Becker A, Theissen G. The major clades of MADS-box genes and their role in the development and evolution of flowering plants[J]. Mol Phylogenet Evol, 2003, 29(3):464-489
[11]	Mandal M A, Yanofsky M. A gene triggering flower formation in Arabidopsis[J]. Nature, 1995, 377(6549):522-524
[12]	Mandel M A, Gustafson-Brown C, Savidge B, et al. Molecular characterization of the Arabidopsis floral homeotic-gene, APETALA1[J]. Nature, 1992, 360(6401):273-277
[13]	Bowman J L, Alvarez J, Weigel D,et al. Control of flower development in Arabidopsis thaliana by APETALA1 and interacting gene[J].Development, 1993, 119(3):721-743
[14]	周兴文, 李纪元, 范正琪, 等. 金花茶查尔酮异构酶基因全长克隆与表达的初步研究[J]. 林业科学研究, 2012,25(1):93-99
[15]	朱高浦, 李纪元, 范正琪, 等. 完全重瓣型山茶花品种‘红十八学士’CjHDEF基因cDNA的序列分析[J]. 生物技术通报, 2011(2):98-102
[16]	朱高浦, 李纪元, 范正琪, 等. 完全重瓣型山茶花品种‘红十八学士’CjHTM6基因cDNA的克隆与分析[J]. 热带作物学报, 2011,32(3):456-462
[17]	朱高浦. 山茶花花MADS-BOX家族B类基因克隆及在重瓣花形成中的作用 [D]. 北京:中国林业科学研究院, 2011:32-36
[18]	邢勇. 猕猴桃科植物的系统发育[J]. 教学与管理, 1987(4):102-104
[19]	Thomas J. Relearning our ABCs: new twists on an old model[J]. Trends Plant Sci, 2001, 6(7):370-376
[20]	赵印泉, 刘青林. 重瓣花的形成机理及遗传特性研究进展[J]. 西北植物学报, 2009, 29(4): 832-841
[21]	中国科学院中国植物志编辑委员会. 中国植物志:第四十九卷第三分册山茶科(一) 山茶亚科[M]. 北京:科学出版社,1998
[22]	Kramer E M, Hall J C. Evolutionary dynamics of genes controlling floral development[J]. Curr Opin Plant Biol, 2005, 8(1):13-18
[23]	Immink R G H, Ferrario S, Busscher-Lange J, et al. Analysis of the petunia MADS-box transcription factor family[J]. Mol Genet Genomics, 2003, 268(5):598-606
[24]	李贵生. 金粟兰花器官特征基因CsAP1、CsAP3和CsSEP3的结构、表达和进化研究 [D]. 北京:中国科学院研究生院, 2005:47-48
[25]	Lowman A C, Purugganan M D. Duplication of the Brassica oleracea APETALA1 floral homeotic gene and the evolution of domesticated cauliflower[J].Hered, 1999, 90(5):514-520
[26]	孙海峰, 孟玉平, 曹秋芬, 等. 枣中APETALA1(AP1)同源基因的克隆及其表达特征分析[J]. 山西大学学报:自然科学版, 2009, 32(2):266-272
[27]	Yao J, Dong Y, Kvarnheden A, et al. Seven MADS-box genes in apple are expressed in different parts of the fruit[J]. J Amer Soc Hort Sci, 1999, 124(1):8-13

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通讯作者: 陈斌, bchen63@163.com

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沈阳化工大学材料科学与工程学院沈阳 110142

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cDNA Cloning and Expression Analysis of CjAPL1 Gene from Camellia japonica

1. Research Institute of Subtropical Forestry, Chinese Academy of Forestry, Fuyang 311400, Zhejiang, China

2. College of Landscape Architecture and Forestry, Qingdao Agricultural University, Qingdao 266109, Shandong, China

Received Date: 2012-11-01

Abstract: In order to study the role of A-class genes in the process of forming plena flower for Camellia japonica, a full-length cDNA sequence of APETALA1 (AP1) gene was cloned from early flower bud of C. japonica ‘Jinpan Lizhi’ using RT-PCR and RACE,named CjAPL1 (GenBank accession # JX657332). The results of nucleotide sequence analysis show that the length of CjAPL1 is 1 149 bp, containing an Opening Reading Frame (ORF) of 741 bp, a 5'-Untranslated Region (5'-UTR) of 206 bp, and a 3'-UTR of 202 bp. The CjAPL1 gene encodes a protein of 246 amino acids, and has more than 75% homology with A-class genes from Actinidia chinensis and Hydrangea macrophylla. Quantitative real-time PCR analysis shows that the gene expression pattern is that in early flower bud of III stage> early flower bud of II stage > early flower bud of I stage > squaring stage, and the tendency shows increasing slowly at the early stage and then decreasing sharply. In different floral organs, the highest expression was observed in style, next in ovary and sepals, and the lowest in bottom stamens and upper outer flowers. The difference of gene expression pattern indicates that CjAPL1 may play a role in the morphogenesis of pleiopetalous flower of Camellia.

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Reference (27)

[1]	Hu J Y, Saedler H. Evolution of the inflated-calyx-syndrome (ICS) in Solanaceae[J]. Mol Biol Evol, 2007, 24(11):2443-2453
[2]	Viaene T, Vekemans D, Irish V F, et al. Pistillata-duplications as a mode for floral diversification in (Basal) asterids[J]. Mol Biol Evol, 2009, 26(11):2627-2645
[3]	Coen E S, Meyerowitz E M. The war of the whorls: Genetic interactions controlling flower development[J]. Nature, 1991, 353(6339):31-37
[4]	Pelaz S, Ditta G S, Baumann E, et al. B and C floral organ identity functions require SEPALLATA MADS-box genes[J].Nature, 2000, 405(6783):200-203
[5]	Theissen G, Saedler H. Floral quartets[J]. Nature, 2001, 409(6819):469-471
[6]	Ditta G, Pinyopich A, Robles P, et al. The SEP4 gene of Arabidopsis thaliana functions in floral organ and meristem identity[J]. Curr Biol, 2004,14(21):1935-1940
[7]	Jack T. Molecular and genetic mechanisms of floral control[J]. Plant Cell, 2004, 16(Suppl):1-17
[8]	徐勇. 桃花发育相关MADS box基因研究 [D].北京:首都师范大学, 2007:23
[9]	Colombo L, Franken J, Koeteje E, et al. The petunia MADS box gene FBP11 determines ovule identity[J]. Plant Cell,1995, 7(11):1859-1868
[10]	Becker A, Theissen G. The major clades of MADS-box genes and their role in the development and evolution of flowering plants[J]. Mol Phylogenet Evol, 2003, 29(3):464-489
[11]	Mandal M A, Yanofsky M. A gene triggering flower formation in Arabidopsis[J]. Nature, 1995, 377(6549):522-524
[12]	Mandel M A, Gustafson-Brown C, Savidge B, et al. Molecular characterization of the Arabidopsis floral homeotic-gene, APETALA1[J]. Nature, 1992, 360(6401):273-277
[13]	Bowman J L, Alvarez J, Weigel D,et al. Control of flower development in Arabidopsis thaliana by APETALA1 and interacting gene[J].Development, 1993, 119(3):721-743
[14]	周兴文, 李纪元, 范正琪, 等. 金花茶查尔酮异构酶基因全长克隆与表达的初步研究[J]. 林业科学研究, 2012,25(1):93-99
[15]	朱高浦, 李纪元, 范正琪, 等. 完全重瓣型山茶花品种‘红十八学士’CjHDEF基因cDNA的序列分析[J]. 生物技术通报, 2011(2):98-102
[16]	朱高浦, 李纪元, 范正琪, 等. 完全重瓣型山茶花品种‘红十八学士’CjHTM6基因cDNA的克隆与分析[J]. 热带作物学报, 2011,32(3):456-462
[17]	朱高浦. 山茶花花MADS-BOX家族B类基因克隆及在重瓣花形成中的作用 [D]. 北京:中国林业科学研究院, 2011:32-36
[18]	邢勇. 猕猴桃科植物的系统发育[J]. 教学与管理, 1987(4):102-104
[19]	Thomas J. Relearning our ABCs: new twists on an old model[J]. Trends Plant Sci, 2001, 6(7):370-376
[20]	赵印泉, 刘青林. 重瓣花的形成机理及遗传特性研究进展[J]. 西北植物学报, 2009, 29(4): 832-841
[21]	中国科学院中国植物志编辑委员会. 中国植物志:第四十九卷第三分册山茶科(一) 山茶亚科[M]. 北京:科学出版社,1998
[22]	Kramer E M, Hall J C. Evolutionary dynamics of genes controlling floral development[J]. Curr Opin Plant Biol, 2005, 8(1):13-18
[23]	Immink R G H, Ferrario S, Busscher-Lange J, et al. Analysis of the petunia MADS-box transcription factor family[J]. Mol Genet Genomics, 2003, 268(5):598-606
[24]	李贵生. 金粟兰花器官特征基因CsAP1、CsAP3和CsSEP3的结构、表达和进化研究 [D]. 北京:中国科学院研究生院, 2005:47-48
[25]	Lowman A C, Purugganan M D. Duplication of the Brassica oleracea APETALA1 floral homeotic gene and the evolution of domesticated cauliflower[J].Hered, 1999, 90(5):514-520
[26]	孙海峰, 孟玉平, 曹秋芬, 等. 枣中APETALA1(AP1)同源基因的克隆及其表达特征分析[J]. 山西大学学报:自然科学版, 2009, 32(2):266-272
[27]	Yao J, Dong Y, Kvarnheden A, et al. Seven MADS-box genes in apple are expressed in different parts of the fruit[J]. J Amer Soc Hort Sci, 1999, 124(1):8-13

cDNA Cloning and Expression Analysis of CjAPL1 Gene from Camellia japonica

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通讯作者: 陈斌, bchen63@163.com

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cDNA Cloning and Expression Analysis of CjAPL1 Gene from Camellia japonica

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