Abstract
Ethylene is one of the most important hormones for flowering of cucumber (Cucumis sativus L.). The molecular interaction between cucumber and ethylene was investigated by construction and characterization of a suppression subtractive hybridization cDNA library from stem apex of ethrel-treated cucumber. To screen differentially expressed genes, dot blotting was used with cDNA probes prepared from mRNAs isolated from control and ethrel-treated samples. Acquired positive clones were subjected to sequence and homology search analysis. In total, 103 expressed sequence tags (ESTs) were acquired after screening; among these, 73 showed significant sequence similarity to known genes, 26 had unknown functions, and four clones did not match any known sequences. Gene ontology (GO) analysis revealed that ESTs with known functions were involved in photosynthesis, metabolism, protein synthesis, cellular communication and transport, signal transduction, and transcriptional regulation. The expression levels and overall profiles in female and male buds of 11 ESTs were confirmed by reverse transcription polymerase chain reaction (RT–PCR) and three of them were detected by real-time quantitative PCR (qRT–PCR) in the whole plant. One of these was an F-box protein, EBF1 (EIN3 BINDING F-BOX1), which might play a positive role in female expression of cucumber.
Similar content being viewed by others
Abbreviations
- EST:
-
Expressed sequence tag
- GO:
-
Gene ontology
- EBF1:
-
EIN3-binding F-box protein 1
- qRT–PCR:
-
Real-time quantitative RT–PCR
- RT–PCR:
-
Reverse transcription polymerase chain reaction
- SSH:
-
Suppression subtractive hybridization
References
Bai SL, Peng YB, Cui JX, Gu HT, Xu LY, Li YQ, Xu ZH, Bai SN (2004) Developmental analyses reveal early arrests of the spore-bearing parts of reproductive organs in unisexual flowers of cucumber (Cucumis sativus L.). Planta 220:230–40
Binder BM, Walker JM, Gagne JM, Emborg TJ, Hemmann G, Bleecker AB, Vierstra RD (2007) The Arabidopsis EIN3 binding F-box protein EBF1 and EBF2 have distinct but overlapping roles in ethylene signaling. Plant Cell 19:509–523
Boualem A, Fergany M, Fernandez R, Troadec C, Martin A, Morin H, Sari MA, Collin F, Flowers JM, Pitrat M, Purugganan MD, Dogimont C, Bendahmane A (2008) A conserved mutation in an ethylene biosynthesis enzyme leads to andromonoecy in melons. Science 321:836–838
Boualem A, Troadec C, Kovalski I, Sari MA, Treves RP, Bendahmane A (2009) A conserved ethylene biosynthesis enzyme leads to andromonoecy in two cucumis species. PLoS One July 3:4(7)
Chan SWL, Henderson IR, Jacobsen SE (2005) Gardening the genome: DNA methylation in Arabidopsis thaliana. Nat Rev Genet 6:351–360
Chen SM, Miao HB, Chen FD, Jiang BB, Lu JG, Fang WM (2009) Analysis of expressed sequence tags (ESTs) collected from the inflorescence of chrysanthemum. Plant Mol Biol Rep 27:503–510
Conaway RC, Brower CS, Conaway JW (2002) Emerging roles of ubiquitin in transcription regulation. Science 296:1254–1258
Daniel AC, Nancy RF, Daniel RG, Daniel MV (2005) Arabidopsis emb 175 and other ppr knockout mutants reveal essential roles for pentatricopeptide repeat (PPR) proteins in plant embryogenesis. Planta 211:424–436
Dellaporta SL, Calderon-Urrea A (1993) Sex determination in flowering plants. Plant Cell 5:1241–1251
Janousek B, Siroky J, Vyskot B (1996) Epigenetic control of sexual phenotype in a dioecious plant, Melandrium album. Mol Gen Genet 250:483–490
Joann ML, Nigel GC, David LR, Schuyler SK (2009) Sequence and in silico characterization of the tomato polygalacturonase (PG) promoter and terminator regions. Plant Mol Biol Rep 27:250–256
Kim WY, Fujiwara S, Suh SS, Kim J, Kim Y, Han L, David K, Putterill J, Nam HG, Somers DE (2007) ZEITLUPE is a circadian photoreceptor stabilized by GIGANTEA in blue light. Nature 449:356–360
Kim HJ, Oh SA, Brownfield L, Hong SH, Ryu H, Hwang I, Twell D, Nam HG (2008) Control of plant germline proliferation by SCFFBL17 degradation of cell cycle inhibitors. Nature 455:1134–1137
Kostov RV, Small CL, McFadden BA (1997) Mutations in a sequence near the N-terminus of the small subunit alter the CO2/O2 specificity factor for ribulose bisphosphate carboxylase/oxygenase. Photosynth Res 54:127–134
Lechner E, Achard P, Vansiri A, Potuschak T, Genschik P (2006) F-box proteins everywhere. Curr Opin Plant Biol 9:631–638
Li Z, Huang SW, Liu SQ, Pan JS, Zhang ZH, Tao QY, Shi QX, Jia ZQ, Zhang WW, Chen HM, Si LT, Zhu LH, Cai R (2009) Molecular isolation of the M gene suggests that a conserved-residue conversion induces the formation of bisexual flowers in cucumber plants. Genetics 182:1381–1385
Liu YZ, Dong T, Lei Y, Deng XX, Gu QQ (2011) Isolation of a polygalacturonase gene from Citrus sinensis fruit and its expression relative to fruit mastication trait, fruit development, and calcium or boron treatments. Plant Mol Biol Rep 29:51–59
Malepszy S, Niemirowicz-Szczytt K (1991) Sex determination in cucumber (Cucumis sativus) as a model system for molecular biology. Plant Sci 80:39–47
Martin A, Troadec C, Boualem A, Rajab M, Fernandez R, Morin H, Pitrat M, Dogimont C, Bendahmane A (2009) A transposon-induced epigenetic change leads to sex determination in melon. Nature 461:1135–1138
Mibus H, Tatlioglu T (2004) Molecular characterization and isolation of the F/f gene for femaleness in cucumber (Cucumis sativus L.). Theor Appl Genet 109:1669–1676
Outchkourov NS, Peters J, de Jong J, Rademakers W, Jongsma MA (2003) The promoter-terminator of chrysanthemum rbcS1 directs very high expression levels in plants. Planta 216:1003–1012
Peñaranda A, Payán MC, Garrido D, Gómez P, Jamilena M (2007) Production of fruits with attached flowers in zucchini squash is correlated with the arrest of maturation of female flowers. J Hortic Sci Biotechnol 82:579–584
Perl-Treves R (1999) Male to female conversion along the cucumber shoot: approaches to studying sex genes and floral development in Cucumis sativus. In: Ainsworth CC (ed) Sex determination in plants. Bios Scientific, Oxford, pp 189–215
Poole CF, Grimball PC (1939) Inheritance of new sex forms in Cucumis melo L. J Hered 30:21–25
Rimon-Knopf R, Trebitsh T (2006) The female-specific Cs-ACS1G gene of cucumber—a case of gene duplication and recombination between the non sex-specific 1-aminocyclopropane-1-carboxylate synthase gene and a branched-chain amino acid transaminase gene. Plant Cell Physiol 47:1217–1228
Roman G, Lubarsky B, Kieber JJ, Rothenberg M, Ecker JR (1995) Genetic analysis of ethylene signal transduction in Arabidopsis thaliana: five novel mutant loci integrated into a stress response pathway. Genetics 139:1393–1409
Saha D, Kumar V, Bhat SR, Srinivasan R (2010) Characterization of upstream sequences of the LOJ gene leads to identification of a novel enhancer element conferring lateral organ junction-specific expression in Arabidopsis thaliana. Plant Mol Biol Rep. doi:10.1007/s11105-010-0229-6
Samach A, Klenz JE, Kohalmi SE, Risseeuw E, Haughn GW, Crosby WL (1999) The UNUSUAL FLORAL ORGANS gene of Arabidopsis thaliana is an F-box protein required for normal patterning and growth in the Xoral meristem. Plant J 20:433–445
Sawa M, Nusinow DA, Kay SA, Imaizumi T (2007) FKF1 and GIGANTEA complex formation is required for day-length measurement in Arabidopsis. Science 318:261–265
Schmitz-Linneweber C, Small I (2008) Pentatricopeptide repeat proteins: a socket set for organelle gene expression. Trends Plant Sci 13(12):663–670
Small ID, Peeters N (2000) The PPR motif—a TPR-related motif prevalent in plant organellar proteins. Trends Biochem Sci 25(2):45–47
Tanaka H, Masuta C, Uehara K, Kataoka J, Koiwai A, Noma M (1997) Morphological changes and hypomethylation of DNA in transgenic tobacco expressing antisense RNA of the s-adenosyl-l-homocysteine hydrolase gene. Plant Mol Biol 35:981–986
Tanurdzic M, Banks JA (2004) Sex-determining mechanisms in land plants. Plant Cell 16:S61–S71
Thines B, Katsir L, Melotto M, Niu Y, Mandaokar A, Liu G, Nomura K, He SY, Howe GA, Browse J (2007) JAZ repressor proteins are targets of the SCFCOI1 complex during jasmonate signalling. Nature 448:661–665
Wang K, Gao F, Zhu RS, Li SQ, Zhu YG (2010) Expression, purification, and secondary structure prediction of pentatricopeptide repeat protein RF1A from rice. Plant Mol Biol Rep. doi:10.1007/s11105-010-0260-7
Xu XB, Liu ZX, Zhang DF, Liu Y, Song WB, Li JS, Dai JR (2009) Isolation and analysis of rice Rf1-orthologus PPR genes co-segregating with Rf3 in maize. Plant Mol Biol Rep 27:511–517
Yamasaki S, Fujii N, Matsuura S, Mizusawa H, Takahashi H (2001) The M locus and ethylene-controlled sex determination in andromonoecious cucumber plants. Plant Cell Physiol 42:608–619
Yamasaki S, Fujii N, Takahashi H (2003) Characterization of ethylene effects on sex determination in cucumber plants. Sex Plant Reprod 16:103–111
Yang ZJ, Peng ZS, Yang H, Yang J, Wei SH, Cai P (2010) Suppression subtractive hybridization identified differentially expressed genes in pistil mutations in wheat. Plant Mol Biol Rep. doi:10.1007/s11105-010-0249-2
Yin T, Quinn JA (1995) Tests of a mechanistic model of one hormone regulating both sexes in Cucumis sativus (Cucurbitaceae). Am J Bot 82:1537–1546
Zhang ZF, Zheng YL (2008) Identification of candidate genes associated with fertility restoration in maize S cytoplasmic male sterility. Plant Mol Biol Rep 26:60–71
Zhou MB, Yang P, Gao PJ, Tang DQ (2011) Identification of differentially expressed sequence tags in rapidly elongating Phyllostachys pubescens internodes by suppressive subtractive hybridization. Plant Mol Biol Rep 29:224–231
Acknowledgments
This work was supported by the Hi-Tech Research and Development Program of China (2007AA10Z177), National Agriculture Science and Technology funding (2010GB2B200131), Heilongjiang Science and Technology Innovation Team (2009td07). This work was carried out in the “Genetic improvement of cold horticultural crops” Key Laboratory of the Ministry of Agriculture of China.
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
ESM 1
(DOC 234 kb)
Rights and permissions
About this article
Cite this article
Jin, X., Qin, Z., Wu, T. et al. Identification of Ethylene-Responsive Genes in Ethrel-Treated Shoot Apices of Cucumber by Suppression Subtractive Hybridization. Plant Mol Biol Rep 29, 875–884 (2011). https://doi.org/10.1007/s11105-011-0304-7
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11105-011-0304-7