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Transgenic pepper plants carrying RNA interference constructs of CaCOI1 gene show severe abnormality

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Abstract

Coronatine insensitive 1 (COI1) is a key regulator in the jasmonate signaling pathway, which plays a central role in jasmonate signaling by forming SCF complexes in the ubiquitination pathway. In this study, the full-length cDNA of CaCOI1 was isolated from chili pepper by reverse transcription PCR. CaCOI1 contains a 1,812-bp open reading frame encoding 603 amino acids. The calculated molecular weight and isoelectric point of the deduced CaCOI1 protein are 68.35 and 6.32 kDa, respectively. CaCOI1 protein shares extensive homology with COI1 proteins of other plant species, and has been predicted to possess a F-box of 30 amino acid residues at the N-terminal and six leucine-rich repeat domains at the C-terminal. Quantitative real-time PCR analysis showed that CaCOI1 can be expressed in detected tissues and organs. The expression level of CaCOI1 in flowers was dramatically increased 2.8–5.4 times higher than that in other tissues and organs. The transgenic suppression of the CaCOI1 gene in the transgenic pepper plants allowed us to test the role of the CaCOI1 protein. CaCOI1-silenced pepper plants showed a severely abnormal phenotype. Compared with wild-type peppers, the CaCOI1-silenced pepper plants suffered from dwarfism with abnormal fruits, and bore misshapen and sterile seeds. The data indicate that CaCOI1 plays an important role in the developmental processes of pepper plants and fruits.

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References

  • Alexandrov NN, Brover VV, Freidin S, Troukhan ME, Tatarinova TV, Zhang H, Swaller TJ, Lu YP, Bouck J, Flavell RB, Feldmann KA (2009) Insights into corn genes derived from large-scale cDNA sequencing. Plant Mol Biol 69:179–194

    Article  PubMed  CAS  Google Scholar 

  • Bennypaul HS, Mutti JS, Rustgi S, Kumar N, Okubara PA, Gill KS (2012) Virus-induced gene silencing (VIGS) of genes expressed in root, leaf, and meiotic tissues of wheat. Funct Integr Genomics 12:143–156

    Article  PubMed  CAS  Google Scholar 

  • Berger S (2001) Jasmonate-related mutants of Arabidopsis as tools for studying stress signaling. Planta 214:497–504

    Article  Google Scholar 

  • Blechert S, Brodschelm W, Hölder S, Kammerer L, Kutchan TM, Mueller MJ, Xia ZQ, Zenk MH (1995) The octadecanoic pathway: signal molecules for the regulation of secondary pathways. Proc Natl Acad Sci USA 92:4099–4105

    Article  PubMed  CAS  Google Scholar 

  • Chini A, Fonseca S, Fernandez G, Adie B, Chico JM, Lorenzo O, Garcia-Casado G, Lopez-Vidriero I, Lozano FM, Ponce MR, Micol JL, Solano R (2007) The JAZ family of repressors is the missing link in jasmonate signalling. Nature 448:666–671

    Article  PubMed  CAS  Google Scholar 

  • Creelman RA, Mullet JE (1997) Biosynthesis and action of jasmonates in plants. Annu Rev Plant Physiol Plant Mol Biol 48:355–381

    Article  PubMed  CAS  Google Scholar 

  • Devoto A, Nieto-Rostro M, Xie DX, Ellis C, Harmston R, Patrick E, Davis J, Sherratt L, Coleman M, Turner JG (2002) COI1 links jasmonate signalling and fertility to the SCF ubiquitin ligase complex in Arabidopsis. Plant J 32:457–466

    Article  PubMed  CAS  Google Scholar 

  • Farmer EE (2001) Surface-to-air signals. Nature 411:854–856

    Article  PubMed  CAS  Google Scholar 

  • Feys BJF, Benedetti CE, Penfold CN, Turner JG (1994) Arabidopsis mutants selected for resistance to the phytotoxin coronatine are male sterile, insensitive to methyl jasmonate, and resistant to a bacterial pathogen. Plant Cell 6:751–759

    PubMed  CAS  Google Scholar 

  • Höfgen R, Willmitzer L (1988) Storage of competent cells for Agrobacterium transformation. Nucleic Acids Res 16:9877

    Article  PubMed  Google Scholar 

  • Howe GA, Jander G (2008) Plant immunity to insect herbivores. Annu Rev Plant Biol 59:41–66

    Article  PubMed  CAS  Google Scholar 

  • Hu T, Wang W, Cao K, Wang X (2006) OsCOI1, a putative COI1 in rice, show MeJA and ABA dependent expression. Prog Biochem Biophys 33:388–393

    CAS  Google Scholar 

  • Hu T, Zeng H, Chen Z, Huang X, Yang Y, Wang G (2012) The optimization of regeneration tissue culture system of three chilli peppers cultivars based on the uniform design and the mathematical model equation. Acta Biol Hung 63:372–388

    Article  PubMed  CAS  Google Scholar 

  • Jaillon O, Aury JM, Noel B, Policriti A, Clepet C, Casagrande A et al (2007) The grapevine genome sequence suggests ancestral hexaploidization in major angiosperm phyla. Nature 449:463–467

    Article  PubMed  CAS  Google Scholar 

  • Katsir L, Schilmiller AL, Staswick PE, He SY, Howe GA (2008) COI1 is a critical component of a receptor for jasmonate and the bacterial virulence factor coronatine. Proc Natl Acad Sci USA 105:7100–7105

    Article  PubMed  CAS  Google Scholar 

  • Kothari SL, Joshi A, Kachhwaha S, Ochoa-Alejo N (2010) Chilli peppers—a review on tissue culture and transgenesis. Biotechnol Adv 28:35–48

    Article  PubMed  CAS  Google Scholar 

  • Lee YH, Kim HS, Kim JY, Jung M, Park YS, Lee JS, Choi SH, Her NH, Lee JH, Hyung NI, Lee CH, Yang SG, Harn CH (2004) A new selection method for pepper transformation: callus-mediated shoot formation. Plant Cell Rep 23:50–58

    PubMed  CAS  Google Scholar 

  • Lee HY, Seo JS, Cho JH, Jung H, Kim JK, Lee JS, Rhee S, Do Choi Y (2013) Oryza sativa COI homologues restore jasmonate signal transduction in Arabidopsis coi1-1 mutants. PLoS One 8:e52802. doi:10.1371/journal.pone.0052802

    Article  PubMed  CAS  Google Scholar 

  • Li L, Zhao Y, McCaig BC, Wingerd BA, Wang J, Whalon ME, Pichersky E, Howe GA (2004) The tomato homolog of coronatine-insensitive1 is required for the maternal control of seed maturation, jasmonate-signaled defense responses, and glandular trichome development. Plant Cell 16:126–143

    Article  PubMed  CAS  Google Scholar 

  • Li C, Zhao J, Jiang H, Wu X, Sun J, Zhang C, Wang X, Lou Y, Li C (2006) The wound response mutant suppressor of prosystemin-mediated responses6 (spr6) is a weak allele of the tomato homolog of coronatine-insensitive1 (COI1). Plant Cell Physiol 47:653–663

    Article  PubMed  CAS  Google Scholar 

  • Lin X, Kaul S, Rounsley S et al (1999) Sequence and analysis of chromosome 2 of the plant Arabidopsis thaliana. Nature 402:761–768

    Article  PubMed  CAS  Google Scholar 

  • Matsumoto T, Tanaka T, Sakai H, Amano N, Kanamori H, Kurita K, Kikuta A, Kamiya K, Yamamoto M, Ikawa H, Fujii N, Hori K, Itoh T, Sato K (2011) Comprehensive sequence analysis of 24,783 barley full-length cDNAs derived from 12 clone libraries. Plant Physiol 156:20–28

    Article  PubMed  CAS  Google Scholar 

  • Melotto M, Mecey C, Niu Y, Chung HS, Katsir L, Yao J, Zeng W, Thines B, Staswick PE, Browse J, Howe GA, He SY (2008) A critical role of two positively charged amino acids in the Jas motif of Arabidopsis JAZ proteins in mediating coronatine- and jasmonoyl isoleucine-dependent interactions with the COI1 F-box protein. Plant J 55:979–988

    Article  PubMed  CAS  Google Scholar 

  • Moon J, Parry G, Estelle M (2004) The ubiquitin–proteasome pathway and plant development. Plant Cell 16:3181–3195

    Article  PubMed  CAS  Google Scholar 

  • Paschold A, Halitschke R, Baldwin IT (2007) Co(i)-ordinating defenses: NaCOI1 mediates herbivore-induced resistance in Nicotiana attenuata and reveals the role of herbivore movement in avoiding defenses. Plant J 51:79–91

    Article  PubMed  CAS  Google Scholar 

  • Paschold A, Bonaventure G, Kant MR, Baldwin IT (2008) Jasmonate perception regulates jasmonate biosynthesis and JA–Ile metabolism: the case of COI1 in Nicotiana attenuata. Plant Cell Physiol 49:1165–1175

    Article  PubMed  CAS  Google Scholar 

  • Paterson AH, Bowers JE, Bruggmann R, Dubchak I, Grimwood J, Gundlach H et al (2009) The Sorghum bicolor genome and the diversification of grasses. Nature 457:551–556

    Article  PubMed  CAS  Google Scholar 

  • Peng SQ, Xu J, Li HL, Tian WM (2009) Cloning and molecular characterization of HbCOI1 from Hevea brasiliensis. Biosci Biotechnol Biochem 73:665–670

    Article  PubMed  CAS  Google Scholar 

  • Ren C, Pan J, Peng W, Genschik P, Hobbie L, Hellmann H, Estelle M, Gao B, Peng J, Sun C, Xie D (2005) Point mutations in Arabidopsis Cullin1 reveal its essential role in jasmonate response. Plant J 42:514–524

    Article  PubMed  CAS  Google Scholar 

  • Reymond P, Farmer EE (1998) Jasmonate and salicylate as global signals for defense gene expression. Curr Opin Plant Biol 1:404–411

    Article  PubMed  CAS  Google Scholar 

  • Sembdner G, Parthier B (1993) The biochemistry and the physiological and molecular actions of jasmonates. Annu Rev Plant Physiol Plant Mol Biol 44:569–589

    Article  CAS  Google Scholar 

  • Shoji T, Ogawa T, Hashimoto T (2008) Jasmonate-induced nicotine formation in tobacco is mediated by tobacco COI1 and JAZ genes. Plant Cell Physiol 49:1003–1012

    Article  PubMed  CAS  Google Scholar 

  • Staswick PE (1992) Jasmonate, genes, and fragrant signals. Plant Physiol 99:804–807

    Article  PubMed  CAS  Google Scholar 

  • Thines B, Katsir L, Melotto M, Niu Y, Mandaokar A, Liu GH, 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

    Article  PubMed  CAS  Google Scholar 

  • Tuskan GA, Difazio S, Jansson S, Bohlmann J, Grigoriev I, Hellsten U et al (2006) The genome of black cottonwood, Populus trichocarpa (Torr. & Gray). Science 313:1596–1604

    Article  PubMed  CAS  Google Scholar 

  • Wang Z, Dai L, Jiang Z, Peng W, Zhang L, Wang G, Xie D (2005) GmCOI1, a soybean F-box protein gene, shows ability to mediate jasmonate-regulated plant defense and fertility in Arabidopsis. Mol Plant Microbe Interact 18:1285–1295

    Article  PubMed  CAS  Google Scholar 

  • Wasternack C, Parthier B (1997) Jasmonate-signalled plant gene expression. Trends Plant Sci 2:302–307

    Article  Google Scholar 

  • Xie DX, Feys BF, James S, Nieto-Rostro M, Turner JG (1998) COI1: an Arabidopsis gene required for jasmonate-regulated defense and fertility. Science 280:1091–1094

    Article  PubMed  CAS  Google Scholar 

  • Xu L, Liu F, Lechner E, Genschik P, Crosby WL, Ma H, Peng W, Huang D, Xie D (2002) The SCFCOI1 ubiquitin–ligase complexes are required for jasmonate response in Arabidopsis. Plant Cell 14:1919–1935

    Article  PubMed  CAS  Google Scholar 

  • Yan Y, Stolz S, Chételat A, Reymond P, Pagni M, Dubugnon L, Farmer EE (2007) A downstream mediator in the growth repression limb of the jasmonate pathway. Plant Cell 19:2470–2783

    Article  PubMed  CAS  Google Scholar 

  • Yan J, Zhang C, Gu M, Bai Z, Zhang W, Qi T, Cheng Z, Peng W, Luo H, Nan F, Wang Z, Xie D (2009) The Arabidopsis coronatine insensitive1 protein is a jasmonate receptor. Plant Cell 21:2220–2236

    Article  PubMed  CAS  Google Scholar 

  • Zhao D, Ma H (2000) Male fertility: a case of enzyme identity. Curr Biol 10:R904–R907

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was funded by the Natural Science Foundation Project of Chongqing Science and Technology Committee (CSTC, 2011jjA80027) and the Natural Science Foundation Project of Chongqing Education Committee (KJ111108).

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Correspondence to Tingzhang Hu.

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Hu, T., Yang, Y., Zeng, H. et al. Transgenic pepper plants carrying RNA interference constructs of CaCOI1 gene show severe abnormality. Mol Breeding 31, 971–979 (2013). https://doi.org/10.1007/s11032-013-9850-z

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