Isolation and Expression of APETALA2 Transcription Factor Gene in Betula platyphylla

ZHANG Yan; LIU Ying; SUN Feng-bin; DAI Chao; LIU Xue-mei

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Isolation and Expression of APETALA2 Transcription Factor Gene in Betula platyphylla

1.
College of Life Science, Northeast Forestry University, Harbin 150040, Heilongjiang, China

Received Date: 2011-07-26

Abstract

AP2 gene is a key transcription factor involved in flower development in plants. The full cDNA of AP2 gene was isolated from Betula platyphylla Suk. by methods of reverse transcription polymerase chain reaction (RT-PCR) and 5’and 3’rapid amplification of cDNA ends (RACE). Results showed that AP2 gene contains an open reading frame (ORF) of 1554bp encoding 517 amino acids. Molecular weight of the deduced protein of BpAP2 is 56.74kDa and the theoretical isoelectric point is 6. 34. The AP2 functional sites and characterized domains were confirmed in the sequence, so the isolated gene was named as BpAP2, and registered in GenBank with accession number JN247408. The deduced amino acid sequence shared 51% 77% of identity with other twelve plant species, the maximum identity with Arabidopsis thaliana (77%) and minimum identity with Pinus thunbergii (51%). A phylogenetic tree was constructed according to multiple sequences alignment of all the thirteen plant species. Transcription expression of BpAP2 was analyzed by qRT-PCR in different tissues and periods in B. platyphylla. Results showed that BpAP2 was more highly expressed in floral organs than in vegetative organs, expression quantity more highly in young tissues than in mature tissues. It inferred that BpAP2 transfactor involved in the regulation of development of floral organs and meristematic tissues in Betula. In addition, a natural male inflorescence- abnormal mutant of B. platyphylla was used for transcription analysis of BpAP2. Results showed that BpAP2 gene is expressed up-regulatedly in female inflorescences, while down-regulation in male inflorescences, young leaves and young shoots, which predicted that BpAP2 should be involved in regulation and expression of multiple genes, and not only be involved in the development of floral organs, but also play some roles in the development of vegetative tissues.
- Betula platyphylla,
- AP2,
- transcription expression,
- qRT-PCR,
- flower development

References

[1]	刘雪梅,杨传平.白桦雌雄花发育周期的时序特征[J].林业科学,2006,42(12):28-33
[2]	刘雪梅,杨传平,耿峰.白桦胚珠发育及胚,胚乳发育关系的研究[J]. 植物研究,2005,25(3):322-326
[3]	Lei Xing,Xue-mei Liu.Characterization of Betula platyphylla gene transcripts associated with early development of male inflorescence [J]. Mol Biol Rep,2011, DOI 10.1007/s11033-011-0818-y
[4]	WEI Jicheng, YANG Chuanping, WANG Chao, et al. Construction and analysis of a subtracted cDNA library of Betula platyphylla female inflorescence[J]. Journal of Forestry Research,2005,16(2):97-100
[5]	闻可心,刘雪梅.AP2功能基因在植物花发育中的重要作用[J].生物技术通报,2010(2):1-7
[6]	Jofuku K Diane, den Boer Bart G W, Van Montagu Marc, et al. Control of Arabidopsis flower and seed development by the homeotic gene APETALA2[J]. The Plant Cell, 1994, 6(9):1211-1225
[7]	Maes Tamara, Van de Steene Nancy, Zethof Jan, et al.Petunia Ap2-like genes and their role in flower and seed development[J]. The Plant Cell, 2001,13(2):244-299
[8]	闻可心.白桦雌雄花差异表达基因的筛选及表达分析.哈尔滨:东北林业大学,2010
[9]	曾凡锁,南楠,詹亚光.富含多糖和次生代谢产物的白桦成熟叶中总的提取[J].植物生理学通讯,2007,43(5):913-916
[10]	戴超,刘雪梅,周菲.白桦基因表达半定量RT-PCR中内参基因的选择[J].经济林研究,2011,29(1):34-39
[11]	Livak K J, Schmittgen T D.Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C (T))[J]. Methods Methods, 2001,25:402–408
[12]	Jing Zhuang, De-Xiang Deng, Quan-Hong Yao, et al. Discovery, phylogeny and expression patterns of AP2-like genes in maize[J].Plant Growth Regul, 2010,62:51-58
[13]	Sowmya Krishnaswamy, Shiv Verma, Muhammad H Rahman, et al. Functional characterization of four APETALA2-family genes (RAP2.6, RAP2.6L, DREB19 and DREB26) in Arabidopsis[J]. Plant Mol Biol, 2011, 75:107-127
[14]	周盛梅,宿红艳,王磊,等.苹果APETALA2同源基因的克隆和转化研究[J].园艺学报,2006,33(2):239-243
[15]	王晨,刘洪,房经贵,等.基于EST数据库的葡萄APETALA2基因cDNA克隆及其表达分析[J].果树学报,2010,27(2):207-212
[16]	刘雪梅,周菲,邢磊,等.白桦雄花突变体及其细胞学特征[J].东北林业大学学报,2010,38(6):1-3,14
[17]	Shigyo Mikao, Ito Motomi. Analysis of gymnosperm two-AP2-domain-containing genes[J]. Development Genes and Evolution, 2004, 214(3): 105-114
[18]	Masa-aki Ohto,Sandra K Floyd, Robert L Fischer, et al. Effects of APETALA2 on embryo, endosperm, and seed coat development determine seed size in Arabidopsis[J]. Sex Plant Reprod,2009,22:277-289
[19]	赵奇,王台,魏小弟.AP2基因在高等植物花器官发育中的作用概述[J].热带农业科学,2005,25(3):50-56
[20]	Irish V F. Floral development in Arabidopsis [J]. Plant Physiol Biochem, 1998, 36 (1-2):61-68
[21]	MA Hao-li, ZHOU Han-lin, ZHANG Huai-yu, et al. Cloning and Expression Analysis of an AP2/ERF Gene and Its Responses to Phytohormones and Abiotic Stresses in Rice[J]. Rice Science, 2010, 17(1): 1-9
[22]	Lifeng Zhao, Shengbao Xu, Tuanyao Chai, et al. OsAP2-1, an AP2-like gene from Oryza sativa, is required for flower development and male fertility[J]. Sex Plant Reprod, 2006,19:197-206
[23]	Zhao Q,Wang T, Wei X D. Function of AP2 gene during floral organs development in higher plant: review[J].Chinese Journal of Tropical Agriculture,2005,259(3):50-56
[24]	Kramer E M, Hall J C. Evolutionary dynamics of genes controlling floral development[J]. Current Opinion in Plant Biology,2005,8:13-18
[25]	Soltis D E, Ma H, Frohlich M W, et al.The floral genome: an evolutionary history of gene duplication and shifting patterns of gene expression[J]. Trends in Plant Science, 2007, 12(8):358-367
[26]	Bomblies K, Dagenais N, Weigel D. Redundant enhancers mediate transcriptional repression of AGAMOUS by APETALA2[J]. Developmental Biology,1999, 216(1): 260-264

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

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Isolation and Expression of APETALA2 Transcription Factor Gene in Betula platyphylla

1. College of Life Science, Northeast Forestry University, Harbin 150040, Heilongjiang, China

Received Date: 2011-07-26

Available Online: 2012-04-13

Abstract: AP2 gene is a key transcription factor involved in flower development in plants. The full cDNA of AP2 gene was isolated from Betula platyphylla Suk. by methods of reverse transcription polymerase chain reaction (RT-PCR) and 5’and 3’rapid amplification of cDNA ends (RACE). Results showed that AP2 gene contains an open reading frame (ORF) of 1554bp encoding 517 amino acids. Molecular weight of the deduced protein of BpAP2 is 56.74kDa and the theoretical isoelectric point is 6. 34. The AP2 functional sites and characterized domains were confirmed in the sequence, so the isolated gene was named as BpAP2, and registered in GenBank with accession number JN247408. The deduced amino acid sequence shared 51% 77% of identity with other twelve plant species, the maximum identity with Arabidopsis thaliana (77%) and minimum identity with Pinus thunbergii (51%). A phylogenetic tree was constructed according to multiple sequences alignment of all the thirteen plant species. Transcription expression of BpAP2 was analyzed by qRT-PCR in different tissues and periods in B. platyphylla. Results showed that BpAP2 was more highly expressed in floral organs than in vegetative organs, expression quantity more highly in young tissues than in mature tissues. It inferred that BpAP2 transfactor involved in the regulation of development of floral organs and meristematic tissues in Betula. In addition, a natural male inflorescence- abnormal mutant of B. platyphylla was used for transcription analysis of BpAP2. Results showed that BpAP2 gene is expressed up-regulatedly in female inflorescences, while down-regulation in male inflorescences, young leaves and young shoots, which predicted that BpAP2 should be involved in regulation and expression of multiple genes, and not only be involved in the development of floral organs, but also play some roles in the development of vegetative tissues.

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Reference (26)

[1]	刘雪梅,杨传平.白桦雌雄花发育周期的时序特征[J].林业科学,2006,42(12):28-33
[2]	刘雪梅,杨传平,耿峰.白桦胚珠发育及胚,胚乳发育关系的研究[J]. 植物研究,2005,25(3):322-326
[3]	Lei Xing,Xue-mei Liu.Characterization of Betula platyphylla gene transcripts associated with early development of male inflorescence [J]. Mol Biol Rep,2011, DOI 10.1007/s11033-011-0818-y
[4]	WEI Jicheng, YANG Chuanping, WANG Chao, et al. Construction and analysis of a subtracted cDNA library of Betula platyphylla female inflorescence[J]. Journal of Forestry Research,2005,16(2):97-100
[5]	闻可心,刘雪梅.AP2功能基因在植物花发育中的重要作用[J].生物技术通报,2010(2):1-7
[6]	Jofuku K Diane, den Boer Bart G W, Van Montagu Marc, et al. Control of Arabidopsis flower and seed development by the homeotic gene APETALA2[J]. The Plant Cell, 1994, 6(9):1211-1225
[7]	Maes Tamara, Van de Steene Nancy, Zethof Jan, et al.Petunia Ap2-like genes and their role in flower and seed development[J]. The Plant Cell, 2001,13(2):244-299
[8]	闻可心.白桦雌雄花差异表达基因的筛选及表达分析.哈尔滨:东北林业大学,2010
[9]	曾凡锁,南楠,詹亚光.富含多糖和次生代谢产物的白桦成熟叶中总的提取[J].植物生理学通讯,2007,43(5):913-916
[10]	戴超,刘雪梅,周菲.白桦基因表达半定量RT-PCR中内参基因的选择[J].经济林研究,2011,29(1):34-39
[11]	Livak K J, Schmittgen T D.Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C (T))[J]. Methods Methods, 2001,25:402–408
[12]	Jing Zhuang, De-Xiang Deng, Quan-Hong Yao, et al. Discovery, phylogeny and expression patterns of AP2-like genes in maize[J].Plant Growth Regul, 2010,62:51-58
[13]	Sowmya Krishnaswamy, Shiv Verma, Muhammad H Rahman, et al. Functional characterization of four APETALA2-family genes (RAP2.6, RAP2.6L, DREB19 and DREB26) in Arabidopsis[J]. Plant Mol Biol, 2011, 75:107-127
[14]	周盛梅,宿红艳,王磊,等.苹果APETALA2同源基因的克隆和转化研究[J].园艺学报,2006,33(2):239-243
[15]	王晨,刘洪,房经贵,等.基于EST数据库的葡萄APETALA2基因cDNA克隆及其表达分析[J].果树学报,2010,27(2):207-212
[16]	刘雪梅,周菲,邢磊,等.白桦雄花突变体及其细胞学特征[J].东北林业大学学报,2010,38(6):1-3,14
[17]	Shigyo Mikao, Ito Motomi. Analysis of gymnosperm two-AP2-domain-containing genes[J]. Development Genes and Evolution, 2004, 214(3): 105-114
[18]	Masa-aki Ohto,Sandra K Floyd, Robert L Fischer, et al. Effects of APETALA2 on embryo, endosperm, and seed coat development determine seed size in Arabidopsis[J]. Sex Plant Reprod,2009,22:277-289
[19]	赵奇,王台,魏小弟.AP2基因在高等植物花器官发育中的作用概述[J].热带农业科学,2005,25(3):50-56
[20]	Irish V F. Floral development in Arabidopsis [J]. Plant Physiol Biochem, 1998, 36 (1-2):61-68
[21]	MA Hao-li, ZHOU Han-lin, ZHANG Huai-yu, et al. Cloning and Expression Analysis of an AP2/ERF Gene and Its Responses to Phytohormones and Abiotic Stresses in Rice[J]. Rice Science, 2010, 17(1): 1-9
[22]	Lifeng Zhao, Shengbao Xu, Tuanyao Chai, et al. OsAP2-1, an AP2-like gene from Oryza sativa, is required for flower development and male fertility[J]. Sex Plant Reprod, 2006,19:197-206
[23]	Zhao Q,Wang T, Wei X D. Function of AP2 gene during floral organs development in higher plant: review[J].Chinese Journal of Tropical Agriculture,2005,259(3):50-56
[24]	Kramer E M, Hall J C. Evolutionary dynamics of genes controlling floral development[J]. Current Opinion in Plant Biology,2005,8:13-18
[25]	Soltis D E, Ma H, Frohlich M W, et al.The floral genome: an evolutionary history of gene duplication and shifting patterns of gene expression[J]. Trends in Plant Science, 2007, 12(8):358-367
[26]	Bomblies K, Dagenais N, Weigel D. Redundant enhancers mediate transcriptional repression of AGAMOUS by APETALA2[J]. Developmental Biology,1999, 216(1): 260-264

Isolation and Expression of APETALA2 Transcription Factor Gene in Betula platyphylla

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通讯作者: 陈斌, bchen63@163.com

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Isolation and Expression of APETALA2 Transcription Factor Gene in Betula platyphylla

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