Welcome to Acta Agronomica Sinica,

Acta Agron Sin ›› 2015, Vol. 41 ›› Issue (06): 972-978.doi: 10.3724/SP.J.1006.2015.00972

• RESEARCH NOTES • Previous Articles     Next Articles

Interaction Analysis of Brassica oleracea L. BoExo70A1 with BoSEC3 and BoExo84 Proteins by Using Yeast Two-hybird System

GAO Qi-Guo1**,LIU Yu-Dong1,**,PU Quan-Ming1,ZHANG Lin-Cheng1,ZHU Li-Quan2,WANG Xiao-Jia1   

  1. 1 College of Horticulture and Landscape Architecture, Southwest University / Key Laboratory of Horticulture Science for Southern Mountainous Regions, Ministry of Education, Chongqing 400716, China; 2 College of Agronomy and Biotechnology, Southwest University, Chongqing 400716, China
  • Received:2014-11-12 Revised:2015-03-19 Online:2015-06-12 Published:2015-04-07


Exo70A1 is necessary for Brassica stigma accepting the compatible pollen, and may play an important role in regulating the movement of water from the dry stigma to the pollen grain and compatible pollen hydration. Here, the coding sequences of BoExo70A1, BoSEC3, BoSEC10, BoSEC15, and BoExo84 genes were amplified from the stigma cDNA of the highly self-incompatible Brassica oleracea L. line A1. Sequence analysis showed that the cDNA sequences of BoExo70A1, BoSEC3, BoSEC10, BoSEC15, and BoExo84 genes were highly homologous to those of AtExo70A1, AtSEC3A, AtSEC10, AtSEC15B, and AtExo84B genes respectively. No signal peptide was found in their deduced protein sequences. BoSEC3 protein contained an EF-hand calcium-binding domain. The BoExo84 protein was divided into two fragments, BoExo84-N and BoExo84-C, and then their encoding sequences were subcloned into pGADT7 vector together with those of BoSEC3, BoSEC10, and BoSEC15 proteins respectively, whereas the encoding sequence of BoExo70A1 protein was subcloned into pGBKT7 vector for analysis of the proteins interaction by yeast two-hybid system. The results showed that BoExo70A1 protein interacted with BoSEC3 or BoExo84-N protein, but not with BoSEC10 or BoSEC15 protein.

Key words: Brassica oleracea L. self-incompatibility, BoExo70A1, Exocyst complex, Interaction

[1]Dickinson H. Dry stigmas, water and self-incompatibility in Brassica. Sex Plant Reprod, 1995, 8: 1–10

[2]Samuel M A, Yee D, Haasen K E, Goring D R. ‘Self’ pollen rejection through the intersection of two cellular pathways in the Brassicaceae: self-compatbility and the compatible pollen response. In: Franklin-Tong V E, ed. Self-incompatibility in flowering plants evolution, diversity, and mechanisms. Berlin: Spring-Verlag, 2008. pp 175–191

[3]Chapman L A, Goring D R. Pollen-pistil interactions regulating successful fertilization in the Brassicaceae. J Exp Bot, 2010, 61: 1987–1999

[4]Iwano M, Takayama S. Self/non-self discrimination in angiosperm self-incompatibility. Curr Opin Plant Biol, 2012, 15: 78–83

[5]Indriolo E, Goring D R. A conserved role for the ARC1 E3 ligase in Brassicaceae self-incompatibility. Front Plant Sci, 2014, 5: 181

[6]Kachroo A, Schopfer C R, Nasrallah M E, Nasrallah J B. Alelle-specific receptor-ligand interactions in Brassica self-incompatibility. Science, 2001, 293: 1824–1826

[7]Haffani Y Z, Gaude T, Cock J M, Goring D R. Antisense suppression of thioredoxin h mRNA in Brassica napus cv. Westar pistil causes a low level constitutive pollen rejection response. Plant Mol Biol, 2004, 55: 619–630

[8]Stone S L, Anderson E M, Mullen R T, Goring D R. ARC1 is an E3 ubiquitin ligase and promotes the ubiquitination of protein during the rejection of self-incompatible Brassica pollen. Plant Cell, 2003, 15: 885–898

[9]Samul M A, Chong Y T, Haasenn K E, Aldea-Brydges M G, Stone S L, Goring D R. Cellular pathways regulating responses to copatible and self-incompatible pollen in Brassica and Arabidopsis stigma intersect at Exo70A1, a putative component of the exocyst complex. Plant Cell, 2009, 21: 2655–2571

[10]Safavian D, Goring D R. Secretory activity is rapidly induced in stigmatic papillae by compatible pollen, but inhibited for self-incompatible pollen in the Brassicaceae. PloS One, 2013, 8: e84286

[11]Safavian D, Jamshed M, Sankaranarayanan S, Indriolo E, Samuel M A, Goring D R. High humidity partially resecues the Arabidopsis thaliana exo70A1 stigmatic defect for accepting compatible pollen. Plant Reprod, 2014, 27: 121–127

[12]Fendrych M, Synek L, Pecenková T, Toupalová H, Cole R, Drdová E, Nebesárová J, Sedinová M, Hála M, Fowler J E, Zársky V. The Arabidopsis exocyst complex is involved in cytokinesis and cell plate maturation. Plant Cell, 2010, 22: 3053–3065

[13]Cvr?ková F, Grunt M, Bezvoda R, Hála M, Kulich I, Rawat A, Zárský V. Evolution of the land plant exocyst complexes. Front Plant Sci, 2012, 3: 159

[14]Hála M, Cole R, Synek L, Drdová E, Pecenková T, Nordheim A, Lamkemeyer T, Madlung J, Hochholdinger F, Fowler J E, Zárský V. An exocyst complex functions in plant cell growth in Arabidopsis and tobacco. Plant Cell, 2008, 20: 1330–1345

[15]Iwano M, Shiba H, Matoba K, Miwa T, Funato M, Entani T, Nakayama P, Shimosato H, Takaoka A, Isogai A, Takayama S. Actin dynamics in papilla cells of Brassica rapa during self- and cross-pollination. Plant Physiol, 2007, 144: 72–81

[16]Finger F P, Hughes T E, Novick P. Sec3p is a spatial landmark for polarized secretion in budding yeast. Cell, 1998, 92: 559–571

[17]Boyd C, Hughes T, Pypaert M, Novick P. Vesicles carry most exocyst subunits to exocytic sites marked by the remaining two subunits, Sec3p and Exo70p. J Cell Biol, 2004, 167: 889–901

[18]Zajac A, Sun X, Zhang J, Guo W. Cyclical regulation of the exocyst and cell polarity determinants for polarized cell growth. Mol Biol Cell, 2005, 16: 1500–1512

[19]Bendezú F O, Vincenzetti V, Martin S G. Fission yeast Sec3 and Exo70 are transported on actin cables and localize the exocyst complexto cell poles. PLoS One, 2012, 7: e40248

[20]Zhang X, Zajac A, Zhang J, Wang P, Li M, Murray J, TerBush D, Guo W. The critical role of Exo84p in the organization and polarized localization of the exocyst complex. J Biol Chem, 2005, 280: 20356–20364

[21]Fendrych M, Synek L, Pecenková T, Drdová E J, Sekeres J, de Rycke R, Nowack M K, Zársky V. Visualization of the exocyst complex dynamics at the plasma membrane of Arabidopsis thaliana. Mol Biol Cell, 2013, 24: 510–520

[1] ZHANG Hai, CHENG Guang-Yuan, YANG Zong-Tao, LIU Shu-Xian, SHANG He-Yang, HUANG Guo-Qiang, XU Jing-Sheng. Sugarcane PsbR subunit response to SCMV infection and its interaction with SCMV-6K2 [J]. Acta Agronomica Sinica, 2021, 47(8): 1522-1530.
[2] WANG Yi-Fan, YIN Wen, HU Fa-Long, FAN Hong, FAN Zhi-Long, ZHAO Cai, YU Ai-Zhong, CHAI Qiang. Response of photosynthetic performance of intercropped wheat to interaction intensity between above- and below-ground [J]. Acta Agronomica Sinica, 2021, 47(5): 929-941.
[3] LI Lan-Lan, MU Dan, YAN Xue, YANG Lu-Ke, LIN Wen-Xiong, FANG Chang-Xun. Effect of OsPAL2;3 in regulation of rice allopathic inhibition on barnyardgrass (Echinochloa crusgalli L.) [J]. Acta Agronomica Sinica, 2021, 47(2): 197-209.
[4] MENG Yu-Yu, WEI Chun-Ru, FAN Run-Qiao, YU Xiu-Mei, WANG Xiao-Dong, ZHAO Wei-Quan, WEI Xin-Yan, KANG Zhen-Sheng, LIU Da-Qun. TaPP2-A13 gene shows induced expression pattern in wheat responses to stresses and interacts with adaptor protein SKP1 from SCF complex [J]. Acta Agronomica Sinica, 2021, 47(2): 224-236.
[5] CHEN Yu-Ting, LIU Lu, CHU Pan-Pan, WEI Jia-Xian, QIAN Hui-Na, CHEN Hua, CAI Tie-Cheng, ZHUANG Wei-Jian, ZHANG Chong. Construction of yeast two-hybrid cDNA library induced by Ralstonia solanacearum and interaction protein screening for AhRRS5 in peanut [J]. Acta Agronomica Sinica, 2021, 47(11): 2134-2146.
[6] JIA Xiao-Ping,YUAN Xi-Lei,LI Jian-Feng,WANG Yong-Fang,ZHANG Xiao-Mei,ZHANG Bo,QUAN Jian-Zhang,DONG Zhi-Ping. Photo-thermal interaction model under different photoperiod-temperature conditions and expression analysis of SiCCT gene in foxtail millet (Setaria italica L.) [J]. Acta Agronomica Sinica, 2020, 46(7): 1052-1062.
[7] ZHENG Qing-Lei,YU Chen-Jing,YAO Kun-Cun,HUANG Ning,QUE You-Xiong,LING Hui,XU Li-Ping. Cloning and expression analysis of sugarcane Fe/S precursor protein gene ScPetC [J]. Acta Agronomica Sinica, 2020, 46(6): 844-857.
[8] Fei-Na ZHENG,Jin-Peng CHU,Xiu ZHANG,Li-Wei FEI,Xing-Long DAI,Ming-Rong HE. Interactive effects of sowing pattern and planting density on grain yield and nitrogen use efficiency in large spike wheat cultivar [J]. Acta Agronomica Sinica, 2020, 46(3): 423-431.
[9] YAO Shu, ZHANG Ya-Dong, LIU Yan-Qing, ZHAO Chun-Fang, ZHOU Li-Hui, CHEN Tao, ZHAO Qing-Yong, ZHU Zhen, Balakrishna Pillay, WANG Cai-Lin. Effects of SSIIa and SSIIIa alleles and their interaction on eating and cooking quality under Wxmp background of rice [J]. Acta Agronomica Sinica, 2020, 46(11): 1690-1702.
[10] DU Jin-Yong,CHAI Qiang,WANG Yi-Fan,FAN Hong,HU Fa-Long,YIN Wen,LI Deng-Ye. Effect of above- and below-ground interaction intensity on photosynthetic characteristics of wheat-maize intercropping [J]. Acta Agronomica Sinica, 2019, 45(9): 1398-1406.
[11] JIA Xiao-Ping,QUAN Jian-Zhang,WANG Yong-Fang,DONG Zhi-Ping,YUAN Xi-Lei,ZHANG Bo,LI Jian-Feng. Effects of different photoperiod conditions on agronomic traits of foxtail millet [J]. Acta Agronomica Sinica, 2019, 45(7): 1119-1127.
[12] Xiao-Qiang ZHAO,Bin REN,Yun-Ling PENG,Ming-Xia XU,Peng FANG,Ze-Long ZHUANG,Jin-Wen ZHANG,Wen-Jing ZENG,Qiao-Hong GAO,Yong-Fu DING,Fen-Qi CHEN. Epistatic and QTL × environment interaction effects for ear related traits in two maize (Zea mays) populations under eight watering environments [J]. Acta Agronomica Sinica, 2019, 45(6): 856-871.
[13] Tao FENG,Chun-Yun GUAN. Cloning and characterization of phytochrome interacting factor 4 (BnaPIF4) gene from Brassica napus L. [J]. Acta Agronomica Sinica, 2019, 45(2): 204-213.
[14] Sha-Sha LI,Geng MA,Wei-Xing LIU,Juan KANG,Yu-Lu CHEN,Yang-Yang HU,Pan-Pan ZHANG,Chen-Yang WANG. Effects of Long-Term Irrigation and Nitrogen Regimes on Soil Nitrogen Content and Paste Property of Wheat Grain [J]. Acta Agronomica Sinica, 2018, 44(7): 1067-1076.
[15] Quan ZHOU, Long-Chang WANG, Shu-Min MA, Xiao-Duan ZHANG, Yi XING, Sai ZHANG. Influences of Rape Intercropping with Chinese Milk Vetch and Straw Mulching on Productive Benefits in Dryland of Southwest China [J]. Acta Agronomica Sinica, 2018, 44(03): 431-441.
Full text



No Suggested Reading articles found!