Cloning and Expression Analysis of CsIAA29 Gene Related to Fruit Development in Cucumber

doi:10.7668/hbnxb.20190916

Abstract

Abstract: Transcriptome analysis was conducted on cucumber short fruit mutants by EMS in the previous experiment,a differentially expressed gene up-regulated in short-fruit mutants was obtained, CsIAA29 (Csa2G381840),log₂FC=2.09. This gene belonged to Aux/IAA family and is involved in auxin response in early plant growth and development. In order to study the role of CsIAA29 gene in the growth and development of cucumber,the CDS sequence of CsIAA29 gene was cloned from cucumber 649 by PCR. Expasy and other online software were used for the physicochemical analysis of CsIAA29 protein. The results showed that CsIAA29 protein encoded 207 amino acids,the molecular formula was C₁₀₆₂H₁₆₃₄N₃₀₂O₃₁₃S₇,the fat coefficient was 71.16,and the total average hydrophilicity was-0.831. Protein structure prediction found four CsIAA29 structure domains,conform to the domain of Aux/IAA family structure characteristics,a preliminary estimate the function of the four structural domain and Aux/IAA family Ⅰ,Ⅱ,Ⅲ and Ⅳ was consistent. Evolutionary analysis showed that CsIAA29 of cucumber was closely related to IAA29 of pumpkin and melon. The results of qRT-PCR showed that the expression of CsIAA29 gene was the highest in cucumber ovary,while the expression of CsIAA29 gene in male flowers and leaves was relatively low. The CsIAA29 gene promoter component was analyzed using the online software PLANTCAT,the results of promoter element analysis showed that the CsIAA29 gene promoter contained several regulatory elements related to the regulation of endogenous growth hormone transport in plants,which were involved in photosynthesis and auxin polarity transport in cucumber. The above experimental results proved that CsIAA29 might affect the growth and development of cucumber fruits by regulating endogenous growth hormone.

Key words: Cucumber, Fruit development, Expression analysis, CsIAA29

CLC Number:

S642.2
X172

YU Yue, CHEN Haiyan, ZHOU Long, GUO Dongxue, GONG Siyu, HAO Ning, DU Yalin, WU Tao. Cloning and Expression Analysis of CsIAA29 Gene Related to Fruit Development in Cucumber[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(S1): 11-17. doi: 10.7668/hbnxb.20190916.

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References

[1] 李亚男,冯霞,陈大清. ARF、Aux/IAA和生长素受体对基因表达的调控[J].安徽农学通报,2008,14(7):36-39. doi:10.16377/j.cnki.issn1007-7731.2008.07.003.
Li Y N,Feng X,Chen D Q. Regulation of gene expression by ARF,Aux/IAA and auxin receptor[J]. Anhui Agricultural Science Bulletin,2008,14(7):36-39.
[2] 王垒,陈劲枫,娄丽娜,娄群峰,贾利. 黄瓜果实中ARF和Aux/IAA基因对外源激素的应答[J].西北植物学报,2011,31(6):1127-1131.
Wang L,Chen J F,Lou L N,Lou Q F,Jia L. Response of ARF and Aux/IAA genes to exogenous hormones in cucumber fruits[J]. Acta Botanica Boreali-occidentalis Sinica,2011,31(6):1127-1131.
[3] Wang L,Lou L N,Yan L Y,Lou Q F,Chen J F. A differential expression analysis of some Aux/IAA family genes in cucumber fruit during the early development[J]. Journal of Nanjing Agricultural University,2011,34(4):156-198.
[4] 傅丰庆. 黄瓜果实发育调控的分子机理研究[D].杭州:浙江大学,2008.
Fu F Q. Molecular mechanism of regulation of cucumber fruit development[D].Hangzhou:Zhejiang University,2008.
[5] Liu K D,Yuan C C,Feng S X,Zong S T,Li H L,Zhong J D,Shen C J,Liu J X. Genome-wide analysis and characterization of Aux/IAA family genes related to fruit ripening in papaya (Carica papaya L.)[J]. BMC Genomics,2017,18(1):892-1024. doi:10.1186/s12864-017-3722-6.
[6] 司马晓娇,郑炳松. 植物生长素原初响应基因Aux/IAA研究进展[J].浙江农林大学学报,2015,32(2):313-318. doi:10.11833/j.issn.2095-0756.2015.02.021.
Sima X J,Zheng B S. Research progress of auxin primary response gene Aux/IAA[J]. Journal of Zhejiang University,2015,32(2):313-318.
[7] Guille T J,Hagen G. Auxin response factors[J]. Current Opinion in Plant Biology,2007,10(5):453-460. doi:10.1016/j.pbi.2007.08.014.
[8] Reed J W. Roles and activities of Aux/IAA proteins in Arabidopsis[J]. Trends Plant Sci,2001,6(9):420-425. doi:10.1016/S1360-1385(01)02042-8.
[9] Sato A,Yama K T. Over-expression of the noncanonical Aux/IAA genes causes auxin-related aberrant phe-notypes in Arabidopsis[J]. Physiol Plant,2008,133(2):397-405. doi:10.1111/j.1399-3054.2008.01055.x.
[10] Middleton A M,King J R,Bennett M J,Owen M R. Mathematical modeling of the Aux/IAA negative feedback loop[J]. Bull Math Biol, 2010,72(6):1383-1407. doi:10.1007/s11538-009-9497-4.
[11] Wang Y J,Deng D X,Bian Y L,Lü Y P,Xie Q. Genome-wide analysis of primary auxin-responsive Aux/IAA gene family in maize(Zea mays L.)[J]. Molecur, 2010,37(8):3991-4001. doi:10.1007/s11033-010-0058-6.
[12] 李小靖. 番茄 SlIAA9、SlIAA14 基因启动子克隆及功能分析[D].海口:海南大学,2010.
Li X J. Cloning and functional analysis of promoters of SlIAA9 and SlIAA14 genes in tomato[D].Haikou:Hainan University,2010.
[13] Wu J,Peng Z,Liu S Y,He Y J,Cheng L,Kong F L,Wang J,Lu G. Genome-wide analysis of Aux/IAA gene family in Solanaceae species using tomato as a model[J]. Mol Genet Genomics,2012,287(4):295-311.doi:10.1007/s00438-012-0675-y.
[14] Atsushi K,Yamashino T,Nakamichi N,Niwa Y,Nakanishi H,Mizuno T. Phytochrome-interacting factor 4 and 5(PIF4 and PIF5) activate the homeobox ATHB2 and auxin-inducible IAA29 gene in the coincidence mechanism underlying photoperiodic control of plant growth of Arabidopsis thaliana[J]. Plant Cell, 2011,52(8):1315-1329.doi:10.1093/pcp/pcr076.
[15] 董秀春,樊金会,于学宁. Aux/IAA基因家族的毛白杨 PtIAA1 的克隆和序列分析[J].分子植物育种,2007,5(4):565-571. doi:10.3969/j.issn.1672-416X.2007.04.020.
Dong X C,Fan J H,Yu X N. Cloning and sequencing of PtIAA1 from populus tomentosa,a member of the Aux/IAA gene family[J]. Molecular Plant Bred,2007,5(4):565-571.
[16] Kalluri U C,Difazio S P,Brunner A M,Tukan G A. Genome-wide analysis of Aux/IAA and ARF gene families in Populus trichocarpa[J]. BMC Plant Biol,2007,7(1):59. doi:10.1186/1471-2229-7-59.
[17] Zhao J Y,Li J,Che C,Pan Y P,Li Y Q,Hou Y,Zhao W S,Zhong Y T,Ding L,Yan S S,Sun C Z,Liu R Y,Yan L Y,Wu T,Li X X,Weng Y X,Zhang X L. A functional allele of CsFUL1 regulates fruit length through repressing CsSUP and inhibiting auxin transport in cucumbert[J].Plant Cell,2019,31(6):1289-1307.doi:10.1105/tpc.18.00905.
[18] Xin T X,Zhang Z,Li S,Zhang S,Li Q,Zhang Z H,Huang S W,Yang X Y. Gentic regulation of ethylene dosage for cucumber fruit elongation[J]. Plant Cell,2019,31(5):1063-1076.doi:10.1105/tpc.18.00957.
[19] Weng Y Q,Colle M,Wang Y H,Yang L M,Rubstion M,Sherman A,Ophir R,Grumet R. QTL mapping in multiple populations and development stages reveals dynamic quantitative trait loci for fruit size in cucumbers of different market classes[J]. Theor Appl Genet,2015,128(9):1747-1763.doi:10.1007/s00122-015-2544-7.
[20] Xu X W,Lu L,Zhu B Y,Xu Q,Qi X H,Chen X H. QTL mapping of cucumber fruit flesh thickness by SLAF-seq[J]. Sci Rep,2015,7(58):158-192. doi:10.1038/srep15829.
[21] 吴铠朝,黄诚梅,李杨瑞,吴建明.TRIzol试剂法快速高效提取3种作物不同组织总RNA[J].甘肃农业大学学报,2012,43(12):1934-1939.doi:10.3969/j:issn.2095-1191.2012.12.1934.
Wu K C,Huang C M,Li Y R,Wu J M. Rapid and efficient extraction of total RNA from different tissues of three crops by TRIzol reagent method[J]. Journal of Gansu Agricultural University, 2012,43(12):1934-1939.
[22] Tom J F. Aux/IAA proteins and auxin signal transduction[J]. Trens Plant Sci,1998,3(6):205-207.doi:10.1016/S1360-1385(98)01244-8.
[23] Kazama H,Dan H,Imaseki H,Wasteneys G. Exposure to ethylene stimulates cell division and alters the fate and polarity of hypocotyl epidermal cells[J]. Plant Physiol,2004,134(4):16141623.doi:10.1104/pp.103.031088.
[24] Wang S C,Hagen G,Guilfoyle T J. Aux/IAA proteins contain a potent transcriptional repression domain[J]. Plant Cell Online,2004,16(2):533-543.doi:10.4161/psb.24526.
[25] 樊书宏,施海涛,刘国银. 一个木薯Aux/IAA转录因子的克隆及调节活性氧功能分析[J].分子植物育种,2019,17(102):362-369.doi:10.13271/j.mpb.017.000362.
Fan S H,Shi H T,Liu G Y. Cloning and functional analysis of a cassava Aux/IAA transcription factor and its regulation of reactive oxygen species[J]. Molecular Plant Breeding,2019,17(102):362-369.
[26] 王垒. 黄瓜单性吉实相关基因的分离、表达及SSR标记分析[D].南京:南京农业大学,2011.
Wang L. Isolation,expression and SSR marker analysis of unisexual gene related to cucumber[D].Nanjing:Nanjing Agricultural University,2011.
[27] 李俊男,燕晓杰,李枢航,张荣沭. 植物AUX/IAA基因家族研究进展[J].中国农学通报,2018,34(15):89-92.doi:10.1111/tpj.12882.
Li J N,Yan X J,Li S H,Zhang R S. Advances in research on AUX/IAA gene family in plants[J]. China Agricultural Science Bulletin,2018,34(15):89-92.
[28] Shinozaki Y,Hao S,Kojima M,Sakakibara H,Ozeki-lida Y,Zheng Y,Fei Z J,Zhong S L,Giovannoni J J,Rose J K C,Okabe Y,Heta Y,Ezura H,Ariizumi T. Ethylene suppresses tomato(Solanuml ycopersicum)fruit set through modification of gibberellin metabolism[J]. Plant Cell, 2015,83(2):237-251. doi:10.1111/tpj.12882.

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