甘蓝BoSPx的克隆与表达分析

doi:10.3864/j.issn.0578-1752.2018.22.011

Abstract

Abstract:

【Objective】Self-incompatibility (SI) is a genetic barrier to inhibit self-pollination and promote hybridization in flowering plants. Here, cloning of a novel gene co-responsive to SI and calcium production during pollination, and spatio-temporal specific expression analysis of the novel gene BoSPx under self-pollination conditions, screening its interaction proteins were conducted to explore. The responding mechanism of BoSPx to self-pollination stimulated stigma in order to provide some further insights into SI process in Brassica oleracea.var.Capitata.【Method】BoSPx was cloned by using transcriptome sequencing, self-pollination and cross-pollination differential screening, and PCR cloning. Amino acid sequence alignment and conserved domain analysis were performed by DNAMAN and Smart software. Expasy online software was used to predict BoSPx protein molecular weight, isoelectric point, secondary structure and transmembrane domain. Phylogenetic tree that constructed by the neighboring method in MEGA6.0 software was used to speculate on the function of BoSPx protein after self-pollination. RT-PCR and qRT-PCR were used to detect BoSPx tissue-specific expression and the relative expression of BoSPx after self-pollination and cross-pollination. The BoSPx-GFP expression vector was constructed and the subcellular localization of BoSPx was observed under confocal microscopy; The interaction proteins were searched by using yeast two-hybrid system. 【Result】A novel gene, which contains a single exon without any introns, named BoSPx was cloned. The open reading frame of BoSPx is 396 bp, encodes a protein with 131 amino acid residues. BoSPx is a hydrophilic protein, no signal peptide and transmembrane, and the theoretical isoelectric point is 4.54. Conserved domains analysis found BoSPx contains three conserved EF-hand motifs (48-60, 64-80, and 81-96). About 500 bp up-stream of BoSPx translation start code contains an auxin response element. RT-PCR analysis found that BoSPx was expressed highest in stigma, BoSPx was also expressed in sepals, leaves, anthers and petals in flowering stage. The BoSPx expression levels in both self-pollinated and cross-pollinated stigma showed “up-down-up” expression pattern. Moreover, The expression of BoSPx in the stigma of self-pollination and cross-pollination increased at first and then decreased down to the highest expression level in stigma in flowering stage. The expression of BoSPx increased rapidly after self-pollination at 15 min and then decreased sharply so that result in SI progress. The decreased value of BoSPx was 1-2 days before flowering. Subcellular location analysis found that BoSPx expression in both the nucleus and cytoplasm. Yeast two-hybrid system did not detect interaction between BoSPx and SRK and ARC1, but BoSPx interacted with auxin family proteins BoSAUR71 and BoPID. 【Conclusion】BoSPx is highly expressed by self-pollination, which may be an auxin-regulated calcium-binding protein, which has a common response to SI and calcium. The protein has multi-tissue expression and nuclear and cytoplasmic properties, indicating that BoSPx may be involved in unknown signaling pathways other than the SRK-ARC1-ExO70A1 pathway.

Key words: Brassica oleracea, BoSPx, yeast two-hybrid, auxin, self-pollination, self-incompatibility

WANG YuKui,BAI XiaoJing,LIAN XiaoPing,ZHANG HeCui,LUO ShaoLan,PU Min,ZUO TongHong,LIU QianYing,ZHU LiQuan. Cloning and Expression Analysis of BoSPx in Brassica oleracea[J].Scientia Agricultura Sinica, 2018, 51(22): 4328-4338.

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References 41

[1]	CHAPMAN L A, GORING D R . Pollen-pistil interactions regulating successful fertilization in the Brassicaceae. Journal of Experimental Botany, 2010,61(7):1987-1999. doi: 10.1093/jxb/erq021 pmid: 20181663
[2]	GU T, MAZZURCO M, SULAMAN WAHEEDA SULAMAN, DINAH D . MATIAS and DAPHNE R.GORING. Binding of an arm repeat protein to the kinase domain of the S-locus receptor kinase. Proceedings of the National Academy of Sciences of the USA, 1998,95(1):382-387. doi: 10.1073/pnas.95.1.382 pmid: 9419384
[3]	STONE S L, ANDERSON M, MULLEN R T, GORING D R . ARC1 is an E3 ubiquitin ligase and promotes the ubiquitination of proteins during the rejection of self-incompatible Brassica pollen. The Plant Cell, 2003,15(4):885-898. doi: 10.1105/tpc.009845 pmid: 12671085
[4]	VANOOSTHUYSE V, TICHTINSKY G, DUMAS C, GAUDE T, MARK C J . Interaction of calmodulin, a sorting nexin and kinase-associated protein phosphatase with the Brassica oleracea S locus receptor kinase. Plant Physiology, 2003,133(2):919-929.
[5]	MIKHAIL E, NASRALL A H, LIU P, JUNE B . Generation of self-incompatible Arabidopsis thaliana by transfer of two S locus genes from A. lyrata. Science, 2002,297:247-249.
[6]	NASRALLAH M E, LIU P, SHERMAN-BROYLES S, BOGGS N A, NASRALLAH J B . Natural variation in expression of self- incompatibility in Arabidopsis thaliana: Implications for the evolution of selfing. Proceedings of the National Academy of Sciences of the USA, 2004,101(45):16070-16074.
[7]	JUNE B, NASRALL A H, MIKHAIL E . Robust self-incompatibility in the absence of a functional ARC1, gene in Arabidopsis thaliana. The Plant Cell, 2014,26:3838-3841.
[8]	Allen A M, Hiscock S J . Evolution and phylogeny of self- incompatibility systems in angiosperms[M] //Self-Incompatibility in Flowering Plants. Springer, Berlin, Heidelberg, 2008: 73-101.
[9]	SAMUEL M A, GORING D R . Self pollen rejection through the intersection of two cellular pathways in the Brassicaceae: Self- incompatibility and the compatible pollen response. Self-Incompatibility in Flowering Plants, 2008: 173-191. doi: 10.1007/978-3-540-68486-2_8
[10]	SAMUEL M A, MUDGIL Y, SALT J N, DELMAS F, RAMACHANDRAN S, CHILELLI A, GORING D R . Interactions between the s-domain receptor kinases and AtPUB-ARM E3 ubiquitin ligases suggest a conserved signaling pathway in Arabidopsis. Plant Physiology, 2008,147:2084-2095. doi: 10.1104/pp.108.123380 pmid: 18552232
[11]	IWANO M, ITO K, FUJII S, KAKITA M, ASANO-SHIMOSATO H, IGARASHI M, KAOTHIEN-NAKAYAMA P, ENTANI T, KANATANI A, TAKEHISA M, TANAKA M, KOMATSU K, SHIBA H, NAGAI T, MIYAWAKI A, ISOGAI A, TAKAYAMA S . Calcium signalling mediates self-incompatibility response in the Brassicaceae. Nature Plants, 2015,1(9):15128.
[12]	ALONI R, ALONI E, LANGHANS M, ULLRICH C I . Role of auxin in regulatingArabidopsis flower development. Planta, 2006(223):315-328.
[13]	齐国辉, 徐继忠, 张玉星 . 鸭梨自交不亲和性与花柱内源激素关系的研究. 河北农业大学学报, 2007,30(1):31-34. doi: 10.3969/j.issn.1000-1573.2007.01.008
	QI G H, XU J Z, ZHANG Y X . Study on the relationship between self-incompatibility of Ya pear and endogenous hormones in style. Journal of Hebei Agricultural University, 2007,30(1):31-34. (in Chinese) doi: 10.3969/j.issn.1000-1573.2007.01.008
[14]	BAVRINA T V, MILYAEVA E L, MACHACCKOVA I . Effect of phytohormone biosynthesis genes (ipt and iaaM+ iaaH) on the sexual reproduction of transgenic tobacco plants. Russian Journal of Plant Physiology, 2002,49(4):484-491. doi: 10.1023/A:1016355824539
[15]	CHEN D, ZHAO J . Free IAA in stigmas and styles during pollen germination. Physiologia Plantarum, 2008,134:202-221. doi: 10.1111/ppl.2008.134.issue-1
[16]	KIM D, PERTEA G, TRAPNELL C, PIMENTEL H, KELLEY R, SALZBERG S L . TopHat2: Accurate alignment of transcriptomes in the presence of insertions, deletions and gene fusions. Genome Biology, 2013,14:R36. doi: 10.1186/gb-2013-14-4-r36
[17]	LIAO X L, ZHU S F, ZHAO W J . Cloning, Sequencing of 16S rDNA of HLB and establishment of real time PCR method. Journal of Agricultural Biotechnology, 2004,12(1):80-85.
[18]	赵娟莹, 刘佳明, 冯志娟 . 大豆锌指转录因子GmDi19-5对高温的响应及互作蛋白的筛选. 中国农业科学, 2017,50(12):2389-2398. doi: 10.3864/j.issn.0578-1752.2017.12.019
	ZHAO J Y, LIU J M, FENG Z J . Response of soybean zinc fingerprinting factor GmDi19-5 to high temperature and screening of interaction proteins. Scientia Agricultura Sinica, 2017,50(12):2389-2398. (in Chinese) doi: 10.3864/j.issn.0578-1752.2017.12.019
[19]	蒲敏, 罗绍兰, 廉小平, 张贺翠, 白晓璟, 王玉奎, 左同鸿, 高启国, 任雪松, 朱利泉 . 自花授粉诱导的甘蓝功能基因BoSPI的克隆与表达分析. 作物学报, 2018,44(2):177-184.
	PU M, LUO S L, LIAN X P, ZHANG H C, BAI X J, WANG Y K, ZUO T H, GAO Q G, REN X S, ZHU L Q . Cloning and expression analysis of bovine functional gene BoSPI induced by self-pollination. Acta Agronomica Sinica, 2018,44(2):177-184. (in Chinese)
[20]	MARSDEN B J, SHAW G S, SYKES B D . Calcium binding proteins. elucidating the contributions to calcium affinity from an analysis of species variants and peptide fragments. Biochemistry and Cell Biology, 1990,68(3):587-601. doi: 10.1139/o90-084 pmid: 2198059
[21]	许俊强 . 甘蓝花粉管钙感受蛋白编码基因CML49的克隆及功能鉴定研究[D]. 重庆: 西南大学, 2014.
	XU J Q . Cloning and functional identification of calcium sensing protein encoding gene CML49 in Brassica oleracea pollen tube[D]. Chongqing: Southwest University, 2014. ( in Chinese)
[22]	Marsden B J, Shaw G S, Sykes B D . Calcium binding proteins. Elucidating the contributions to calcium affinity from an analysis of species variants and peptide fragments. Biochemistry and Cell Biology, 1990,68(3):587-601. doi: 10.1139/o90-084 pmid: 2198059
[23]	GOH C S, LEE Y, KIM S H . Calcium could be involved in auxin- regulated maintenance of the quiescent center in the Arabidopsis root. Journal of Plant Biology, 2012,55(2):143-150. doi: 10.1007/s12374-011-9197-0
[24]	VANNESTE S, FRIML J . Calcium: The missing link in auxin action. Plants, 2013,2:650-675. doi: 10.3390/plants2040650
[25]	TANTIKANJANA T, NASRALLAH J B . Non-cell-autonomous regulation of crucifer self-incompatibility by auxin response factor ARF3. Proceedings of the National Academy of Sciences of the USA, 2012,109(47):19468-19473. doi: 10.1073/pnas.1217343109 pmid: 23129621
[26]	毕云龙, 高启国, 施松梅, 刘晓欢, 蒲全明, 张莹, 张林成, 廉小平, 柳菁, 朱利泉 . 甘蓝BoSU03蛋白基因的克隆及其与SRK相互作用分析. 园艺学报, 2015,42(11):2206-2214. doi: 10.16420/j.issn.0513-353x.2015-0352
	BI Y L, GAO Q G, SHI S M, LIU X H, PU Q M, ZHANG Y, ZHANG L C, LIAN X P, LIU J, ZHU L Q . Cloning of BoSU03 protein gene and its interaction with SRK. Chinese Journal of Horticulture, 2015,42(11):2206-2214. (in Chinese) doi: 10.16420/j.issn.0513-353x.2015-0352
[27]	THOMAS S G, HUANG S, LI S, STAIGER C J, VERNONICA E, FRANKLIN T . Actin depolymerization is sufficient to induce programmed cell death in self-incompatible pollen. The Journal of Cell Biology, 2006,174(2):221-229. doi: 10.1083/jcb.200604011 pmid: 16831890
[28]	GEITMANN A, SNOWMAN B N, EMONS A M C, VERNONICA E, FRANKLIN T . Alterations in the actin cytoskeleton of pollen tubes are induced by the self-incompatibility reaction in Papaver rhoeas. The Plant Cell, 2000,12(7):1239-1251.
[29]	CHEUNG A Y . Pollen-pistil interactions during pollen-tube growth. Trends in Plant Science, 1996,1(2):45-51. doi: 10.1016/S1360-1385(96)80028-8
[30]	HISCOCK S J, ALLEN A M . Diverse cell signalling pathways regulate pollen-stigma interactions: The search for consensus. New Phytologist, 2008,179(2):286-317. doi: 10.1111/j.1469-8137.2008.02457.x
[31]	SHI S, GAO Q, ZENG J, LIU X H, PU Q M, ZHANG H C, YANG X H, ZHU L Q . N-terminal domains of ARC1 are essential for interaction with the N-terminal region of Exo70A1 in transducing self-incompatibility of Brassica oleracea. Acta Biochimica et Biophysica Sinica, 2016,48(9):777-787. doi: 10.1093/abbs/gmw075
[32]	杨红, 朱利泉, 张贺翠 . 利用酵母双杂交系统鉴定甘蓝SCR与SRK胞外域片段间的相互作用. 中国农业科学, 2011,44(9):1953-1962. doi: 10.3864/j.issn.0578-1752.2011.09.024
	YANG H, ZHU L Q, ZHANG H C . Study on the interactions between the truncated fragments of SCR and eSRK from Brassica oleracea L. by a yeast two-hybrid system. Scientia Agricultura Sinica, 2011,44(9):1953-1962. (in Chinese) doi: 10.3864/j.issn.0578-1752.2011.09.024
[33]	施松梅, 高启国, 廉小平, 毕云龙, 刘晓欢, 蒲全明, 刘贵喜, 柳菁, 任雪松, 杨晓红, 朱利泉, 王小佳 . 结球甘蓝SRK-ARC1-Exo70A1互作域的确定及作用强度. 中国农业科学, 2016,49(1):14-26. doi: 10.3864/j.issn.0578-1752.2016.01.002
	SHI S M, GAO Q G, LIAN X P, BI Y L, LIU X H, PU Q M, LIU G X, LIU J, REN X S, YANG X H, ZHU L Q, WANG X J . Determination and interaction intensity of the SRK-ARC1-Exo70A1 Interaction field of Brassica oleracea. Scientia Agricultura Sinica, 2016,49(1):14-26. (in Chinese) doi: 10.3864/j.issn.0578-1752.2016.01.002
[34]	张贺翠, 柳菁, 廉小平, 曾静, 杨昆 . 甘蓝 ROH1与EXO70A1的表达与相互作用. 中国农业科学, 2016,49(4):775-783. doi: 10.3864/j.issn.0578-1752.2016.04.016
	ZHANG H C, LIU J, LIAN X P, ZENG J, YANG K . Expression and Interaction of ROH1 and EXO70A1 in Brassica oleracea. Scientia Agricultura Sinica, 2016,49(4):775-783. (in Chinese) doi: 10.3864/j.issn.0578-1752.2016.04.016
[35]	BENJAMINS R, CARLOS S, AMPUDIA G, HOOYKAAS P J J, OFFRINGA R . PINOID-mediated signaling involves calcium-binding proteins. Plant Physiology, 2003,132(3):1623-1630. doi: 10.1104/pp.103.019943
[36]	CHRISTENSEN S K, DAGENAIS N, CHORY J, WEIGEL D . Regulation of auxin response by the protein kinase PINOID. Cell, 2000,100(4):469-478. doi: 10.1016/S0092-8674(00)80682-0 pmid: 10693763
[37]	FRIML J, YANG X, MICHNIEWICZ M, WEIJERS D, QUINT A, TIETZ O . A PINOID-dependent binary switch in apical-basal PIN polar targeting directs auxin efflux. Science, 2004,306(5697):862-865. doi: 10.1126/science.1100618
[38]	ZEGZOUTI H, ANTHONY R G, JAHCHAN N, BOGRE L, CHRISTENSEN S K . Phosphorylation and activation of PINOID by the phospholipid signaling kinase 3-phosphoinositide-dependent protein kinase 1 (PDK1) in Arabidopsis. Proceedings of the National Academy of Sciences of the USA, 2006,103(16):6404-6409. doi: 10.1073/pnas.0510283103 pmid: 16601102
[39]	MICHNIEWICZ M, ZAGO M K, ABAS L, WEIJERS D, SCHWEIGHOFER A, MESKIENE I, HEISLER M, GOHNO C, ZHANG J, HUANG F, SCHWAB R, WEIGEL D, MEYEROWITZ E M, LUSCHNIG C, OFFRINGA R, FRIML J . Antagonistic regulation of PIN phosphorylation by PP2A and PINOID directs auxin flux. Cell, 2007,130(6):1044-1056. doi: 10.1016/j.cell.2007.07.033 pmid: 17889649
[40]	PARK J E, KIM Y S, YOON H K, PARK C M . Functional characterization of a small auxin-up RNA gene in apical hook development in Arabidopsis. Plant Science, 2007,172(1):150-157. doi: 10.1016/j.plantsci.2006.08.005
[41]	KANT S, BI Y M, ZHU T, ROTHSTEIN S J . SAUR39, a small auxin-up RNA gene, acts as a negative regulator of auxin synthesis and transport in rice. Plant Physiology, 2009,151(2):691-701.

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相关功能预测 Associated putative function	启动子顺式作用元件 Cis-elements in the promoter region
脱落酸响应元件Abscisic acid responsiveness	ABRE
光响应元件Light responsive element	ATCT-motif, Box4, G-Box, ACE, CATT-motif, GAG-motif Gap-box, TCCC-motif, I-box, MNF1, TCT- motif, as-2-box
顺式作用调节元件Cis-acting regulatory element	ARE
启动子和增强子区域Promoter and enhancer regions	CAAT-box
热应激反应Heat stress responsiveness	HSE
低温响应元件Low-temperature responsiveness	LTR
干旱诱导反应元件Drought induced response element	MBS
调控胚乳表达元件Regulatory element for endosperm expression	GCN4_motif, Skn-1_motif
水杨酸响应元件Salicylic acid responsiveness	TCA-element
茉莉酸响应元件MeJA-responsiveness	CGTCA-motif, TGACG-motif
生长素响应元件Auxin-responsive element	TGA-element

编号 No.	酵母菌种（质粒） Mating strain (plasmid)	培养基 Yeast medium	菌斑 Colony	颜色 Color
1	Y2HGold（pGADT7-T×pGBKT7-53）	SD/-Leu/-Trp	是Yes	白色White
2	Y2HGold(pGADT7-T×pGBKT7-Lam)	SD/-Leu/-Trp	是Yes	红色Red
3	Y2HGold (pGADT7-SRK×pGBKT7-BoSPx)	SD/-Leu/-Trp	是Yes	红色Red
4	Y2HGold (pGADT7-ARC1×pGBKT7-BoSPx)	SD/-Leu/-Trp	是Yes	红色Red
5	Y2HGold (pGADT7-SAUR71×pGBKT7-BoSPx)	SD/-Leu/-Trp	是Yes	白色White
6	Y2HGold (pGADT7-BoPID×pGBKT7-BoSPx)	SD/-Leu/-Trp	是Yes	白色White
7	Y2HGold（pGADT7-T×pGBKT7-53）	SD/-Ade/-His/-Leu/-Trp	是Yes	白色White
8	Y2HGold(pGADT7-T×pGBKT7-Lam)	SD/-Ade/-His/-Leu/-Trp	是Yes	红色Red
9	Y2HGold (pGADT7-SRK×pGBKT7-BoSPx)	SD/-Ade/-His/-Leu/-Trp	无No	无No
10	Y2HGold (pGADT7-ARC1×pGBKT7-BoSPx)	SD/-Ade/-His/-Leu/-Trp	无No	无No
11	Y2HGold (pGADT7-SAUR71×pGBKT7-BoSPx)	SD/-Ade/-His/-Leu/-Trp	是Yes	白色White
12	Y2HGold (pGADT7-BoPID×pGBKT7-BoSPx)	SD/-Ade/-His/-Leu/-Trp	是Yes	白色White

Cloning and Expression Analysis of BoSPx in Brassica oleracea

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