Molecular Cloning, Expression Analysis and Localization of Exo70A1 Related to Self Incompatibility in Non-Heading Chinese Cabbage (Brassica campestris ssp. chinensis)

doi:10.1016/S2095-3119(13)60580-8

Journal of Integrative Agriculture ›› 2013, Vol. 12 ›› Issue (12): 2149-2156.DOI: 10.1016/S2095-3119(13)60580-8

Molecular Cloning, Expression Analysis and Localization of Exo70A1 Related to Self Incompatibility in Non-Heading Chinese Cabbage (Brassica campestris ssp. chinensis)

WANG Li, GE Ting-ting, PENG Hai-tao, WANG Cheng, LIU Tong-kun, HOU Xi-lin

1. Horticultural Department, Nanjing Agricultural University, Nanjing 210095, P.R.China
2.State Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing 210095, P.R.China
3.Key Laboratory of Southern Vegetable Crop Genetic Improvement, Ministry of Agriculture, Nanjing 210095, P.R.China

收稿日期:2012-11-19 出版日期:2013-12-01 发布日期:2013-12-17
通讯作者: LI Ying, Tel: +86-25-84395756, Fax: +86-25-84395266, E-mail: yingli@njau.edu.cn
作者简介:WANG Li, Tel: +86-25-84395756, Fax: +86-25-84395266, E-mail: 2010204017@njau.edu.cn
基金资助:
This work is supported by the National Basic Research Program of China (973 Program, 2009CB119001), the Fundamental Research Funds for the Central Universities of China (KYZ201111), the Natural Science Foundation of Jiangsu Province, China (BK2011643), and the Scientific Innovation Research of College Graduate of Jiangsu Province, China (CXLX11_0688).

Molecular Cloning, Expression Analysis and Localization of Exo70A1 Related to Self Incompatibility in Non-Heading Chinese Cabbage (Brassica campestris ssp. chinensis)

WANG Li, GE Ting-ting, PENG Hai-tao, WANG Cheng, LIU Tong-kun, HOU Xi-lin

1. Horticultural Department, Nanjing Agricultural University, Nanjing 210095, P.R.China
2.State Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing 210095, P.R.China
3.Key Laboratory of Southern Vegetable Crop Genetic Improvement, Ministry of Agriculture, Nanjing 210095, P.R.China

Received:2012-11-19 Online:2013-12-01 Published:2013-12-17
Contact: LI Ying, Tel: +86-25-84395756, Fax: +86-25-84395266, E-mail: yingli@njau.edu.cn
About author:WANG Li, Tel: +86-25-84395756, Fax: +86-25-84395266, E-mail: 2010204017@njau.edu.cn
Supported by:
This work is supported by the National Basic Research Program of China (973 Program, 2009CB119001), the Fundamental Research Funds for the Central Universities of China (KYZ201111), the Natural Science Foundation of Jiangsu Province, China (BK2011643), and the Scientific Innovation Research of College Graduate of Jiangsu Province, China (CXLX11_0688).

摘要/Abstract

摘要： The exocyst is a conserved protein complex, and required for vesicles tethering, fusion and polarized exocytosis. Exo70A1, the exocyst subunit, is essential for assembly of the exocyst complex. To better understand potential roles of Exo70A1 in non-heading Chinese cabbage (Brassica campestris ssp. chinensis), we obtained the full-length cDNA of Exo70A1 gene, which consisted of 1 917 bp and encoded a protein of 638 amino acids. BlastX showed BcExo70A1 shared 94.9% identity with Brassica oleracea var. acephala (AEI26267.1), and clustered into a same group with other homologues in B. oleracea var. acephala and Brassica napus. Subcellular localization analysis showed BcExo70A1 was localized to punctate structures in cytosol of onion epithelial cells. Results showed that BcExo70A1 was widely presented in stamens, young stems, petals, unpollinated pistils, roots and leaves of self compatible and incompatible plants. The transcripts of BcExo70A1 in non- heading Chinese cabbage declined during initial 1.5 h after incompatible pollination, while an opposite trend was presented after compatible pollination. Our study reveals that BcExo70A1 could play essential roles in plant growth and development, and is related to the rejection of self pollen in non-heading Chinese cabbage.

关键词: Exo70A1 , gene expression , subcellular localization , self-incompatibility , non-heading Chinese cabbage

Abstract: The exocyst is a conserved protein complex, and required for vesicles tethering, fusion and polarized exocytosis. Exo70A1, the exocyst subunit, is essential for assembly of the exocyst complex. To better understand potential roles of Exo70A1 in non-heading Chinese cabbage (Brassica campestris ssp. chinensis), we obtained the full-length cDNA of Exo70A1 gene, which consisted of 1 917 bp and encoded a protein of 638 amino acids. BlastX showed BcExo70A1 shared 94.9% identity with Brassica oleracea var. acephala (AEI26267.1), and clustered into a same group with other homologues in B. oleracea var. acephala and Brassica napus. Subcellular localization analysis showed BcExo70A1 was localized to punctate structures in cytosol of onion epithelial cells. Results showed that BcExo70A1 was widely presented in stamens, young stems, petals, unpollinated pistils, roots and leaves of self compatible and incompatible plants. The transcripts of BcExo70A1 in non- heading Chinese cabbage declined during initial 1.5 h after incompatible pollination, while an opposite trend was presented after compatible pollination. Our study reveals that BcExo70A1 could play essential roles in plant growth and development, and is related to the rejection of self pollen in non-heading Chinese cabbage.

Key words: Exo70A1 , gene expression , subcellular localization , self-incompatibility , non-heading Chinese cabbage

WANG Li, GE Ting-ting, PENG Hai-tao, WANG Cheng, LIU Tong-kun, HOU Xi-lin. Molecular Cloning, Expression Analysis and Localization of Exo70A1 Related to Self Incompatibility in Non-Heading Chinese Cabbage (Brassica campestris ssp. chinensis)[J]. Journal of Integrative Agriculture, 2013, 12(12): 2149-2156.

参考文献

[1]Boyd C, Hughes T, Pypaert M, Novick P. 2004. Vesicles carry most exocyst subunits to exocytic sites marked by the remaining two subunits, Sec3p and Exo70p. The Journal of Cell Biology, 167, 889-901

[2]Cao S C, Hou X L, Hao X M. 2002. Studies on breeding and seed reproduction technique of Aijiaohuang self- incompatible line in non-heading Chinese cabbage. Journal of Nanjing Agricultural University, 25, 111-113 (in Chinese)

[3]Chong Y T, Gidda S K, Sanford C, Parkinson J, Mullen R T, Goring D R. 2010. Characterization of the Arabidopsis thaliana exocyst complex gene families by phylogenetic, expression profiling, and subcellular localization studies. New Phytologist, 185, 401-419

[4]Elias M, Drdova E, Ziak D, Bavlnka B, Hala M, Cvrckova F, Soukupova H, Zarsky V. 2003. The exocyst complex in plants. Cell Biology International, 27, 199-201

[5]Finger F P, Hughes T E, Novick P. 1998. Sec3p is a spatial landmark for polarised secretion in budding yeast. Cell, 92, 559-571

[6]Guo W, Roth D, Walch-Solimena C, Novick P. 1999. The exocyst is an effector for Sec4p, targeting secretory vesicles to sites of exocytosis. The EMBO Journal, 4, 71-80

[7]Hála M, Cole R, Synek L, Drdová E, Pecenková T, Nordheim A, Lamkemeyer T, Madlung J, Hochholdinger F, Fowler J E, et al. 2008. An exocyst complex functions in plant cell growth in Arabidopsis and tobacco. The Plant Cell, 20, 1330-1345

[8]He B, Guo W. 2009. The exocyst complex in polarized exocytosis. Current Opinion in Cell Biology, 21, 537-542

[9]Hsu S C, Ting A E, Hazuka C D, Davanger S, Kenny J W, Kee Y, Scheller R H. 1996. The mammalian brain rsec6/8 complex. Neuron, 17, 1209-1219

[10]Hsu S C, TerBush D, Abraham M, Guo W. 2004. The exocyst complex in polarised exocytosis. International Review of Cytology, 233, 243-265

[11]Ivanov R, Fobis-Loisy I, Gaude T. 2010. When no means no: guide to Brassicaceae self-incompatibility. Trends in Plant Science, 15, 387-394

[12]Iwano M, Shiba H, Matoba K, Miwa T, Funato M, Entani T, Nakayama P, Shimosato H, Takaoka A, Isogai A, et al. 2007. Actin dynamics in papilla cells of Brassica rapa during self- and cross-pollination. Plant Physiology, 144, 72-81

[13]Kee Y, Yoo J S, Hazuka C D, Peterson K E, Hsu S C, Scheller R H. 1997. Subunit structure of the mammalian exocyst complex. Proceedings of the National Academy of Sciences of the United States of America, 94, 14438- 14443.

[14]Li S, van Os G M, Ren S, Yu D, Ketelaar T, Emons A M, Liu C M. 2010. Expression and functional analyses of EXO70 genes in Arabidopsis implicate their roles in regulating cell type-specific exocytosis. Plant Physiology, 154, 1819-1830

[15]Livak K J, Schmittgen T D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2(T)(-Delta Delta C) method. Methods, 25, 402-408

[16]Munson M, Novick P. 2006. The exocyst defrocked, a framework of rods revealed. Nature Structural & Molecular Biology, 13, 577-581

[17]Samuel M A, Chong Y T, Haasen K E, Aldea-Brydges M G, Stone S L, Goring D R. 2009. Cellular pathways regulating responses to compatible and self-incompatible pollen in Brassica and Arabidopsis stigmas intersect at Exo70A1, a putative component of the exocyst complex. The Plant Cell, 21, 2655-2671

[18]Samuel M A, Tang W, Jamshed M, Northey J, Patel D, Smith D, Siu K W, Muench D G, Wang Z Y, Goring D R. 2011. Proteomic analysis of Brassica stigmatic proteins following the self-incompatibility reaction reveals a role for microtubule dynamics during pollen responses. Molecular & Cellular Proteomics, 10. doi: 10.1074/mcp. M111.011338

[19]Sheen J, Hwang S, Niwa Y, Kobayashi H, Galbraith D W. 1995. Green fluorescent protein as a new vital marker in plant cells. Plant Journal, 8, 777-784

[20]Synek L, Schlager N, Elia´s M, Quentin M, Hauser M T, Za´rsky´ V. 2006. AtEXO70A1, a member of a family of putative exocyst subunits specifically expanded in land plants, is important for polar growth and plant development. Plant Journal, 48, 54-72

[21]Tamura K, Dudley J, Nei M, Kumar S. 2007. MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Molecular Biology and Evolution, 24, 1596-1599

[22]TerBush D R, Maurice T, Roth D, Novick P. 1996. The exocyst is a multiprotein complex required for exocytosis in Saccharomyces cerevisiae. The EMBO Journal, 15, 6483-6494

[23]TerBush D R, Novick P. 1995. Sec6, Sec8, and Sec15 are components of a multisubunit complex which localizes to small bud tips in Saccharomyces cerevisiae. The Journal of Cell Biology, 130, 299-312

[24]Tsuboi T, Ravier M A, Xie H, Ewart M A, Gould G W, Baldwin S A, Rutter G A. 2005. Mammalian exocyst complex is required for the docking step of insulin vesicle exocytosis. The Journal of Biological Chemistry, 280, 25565-25570

[25]Xiao D, Zhang N W, Zhao J J, Bonnema G, Hou X L. 2012. Validation of reference genes for real-time quantitative PCR normalisation in non-heading Chinese cabbage. Functional Plant Biology, 39, 342-350

[26]Wang L, Hou X L, Zhang A F, Li Y. 2012. Effect of NaCl on overcoming self-incompatibility in non-heading Chinese cabbage (Brassica campestris ssp. chinensis Makino) studied by fluorescent microscopy. Acta Horticulturae, 932, 127-132

[27]Yang K, Zhou Y X, Zhang H C, Zhao Y B, Yang Y J, Lu J X, Zhu L Q, Xue L Y, Lv J, Gao Q G. 2012. Cloning and expression characteristics of EX070A1 from Brassica oleracea, Brassica campestris, and Brassica napus. Acta Agronomica Sinica, 38, 578-588 (in Chinese)

Molecular Cloning, Expression Analysis and Localization of Exo70A1 Related to Self Incompatibility in Non-Heading Chinese Cabbage (Brassica campestris ssp. chinensis)

Molecular Cloning, Expression Analysis and Localization of Exo70A1 Related to Self Incompatibility in Non-Heading Chinese Cabbage (Brassica campestris ssp. chinensis)

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