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Expression of a Chinese cabbage cysteine proteinase inhibitor, BrCYS1, retards seed germination and plant growth in transgenic Tobacco plant

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Abstract

Phytocystatins are plant cysteine proteinase inhibitors that regulate endogenous and heterologous cysteine proteinases of the papain family. A cDNA encoding the phytocystatin BrCYS1 (Brassica rapa cysteine proteinase inhibitor 1 ) has been isolated from Chinese cabbage (B. rapa subsp.pekinensis) flower buds. In order to explore the role of this inhibitory enzyme, tobacco plants (Nicotiana tabacum L. cv. Samson) containing altered amounts of phytocystatin were generated by over-expressingBrCYS1 cDNA in either the sense or the antisense configuration. The resulting plants hadin vitro enzyme inhibitory activities that were over 10% of those detected in wild type plants. The transgenic plants exhibited retarded seed germination and seedling growth and a reduced seed yield, whereas these properties were enhanced in antisense plants. These data suggest that BrCYS1 participates in the control of seed germination, post-germination and plant growth by regulating cysteine peptidase activity.

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Literature Cited

  • Abe K, Emori Y, Kondo H, Suzuki K, Arai S (1987) Molecular cloning of a cysteine proteinase inhibitor of rice (Oryzacystatin). J Biol Chem 262: 16793–16797

    PubMed  CAS  Google Scholar 

  • Abe K, Kondo H, Watanabe H, Emori Y, Arai S (1991) Oryzacystatins as the first well-defined cystatins of plant origin and their target proteinases in rice seeds. Biomed Biochim Acta 50: 637–641

    PubMed  CAS  Google Scholar 

  • Abe M, Abe K, Iwabuchi K, Domoto C, Arai S (1994) Corn cystatin I expressed in Escherichia coli: investigation of its inhibitory profile and occurrence in corn kernels. J Biochem 116: 488–492

    PubMed  CAS  Google Scholar 

  • Abe M, Abe K, Kuroda M, Arai S (1992) Corn kernel cysteine proteinase inhibitor as a novel cystatin superfamily member of plant origin. Eur J Biochem 209: 933–937

    Article  PubMed  CAS  Google Scholar 

  • Barrett AJ (1987) The cystatins: a new class of peptidase inhibitors. Trends Biochem Sci 12: 193–196

    Article  CAS  Google Scholar 

  • Benchekroun A, Michaud D, Nguyen-Quoc B, Overnay S, Desjardins Y, Yelle S (1995) Synthesis of active oryzacystatin I in transgenic potato plants. Plant Cell Reports 14: 585–588

    Article  CAS  Google Scholar 

  • Bonade-Bottino M, Lerin J, Zaccomer B, Jouanin L (1999) Physiological adaptation explains the insensitivity ofBaris coerulescens to transgenic oilseed rape expressing oryzacystatin I. Insect Biochem Mol Biol 29: 131–138

    Article  CAS  Google Scholar 

  • Botella MA, Xu Y, Prabha TN, Zhao Y, Narasimhan ML, Wilson KA, Nielsen SS, Bressan RA, Hasegawa PM (1996) Differential expression of soybean cysteine proteinase inhibitor genes during development and in response to wounding and methyl jasmonate. Plant Physiol 112: 1201–1210

    Article  PubMed  CAS  Google Scholar 

  • Debeaujon I, Léon-Kloosterziel KM, Koornneef M (2000) Influence of the testa on seed dormancy, germination, and longevity inArabidopsis. Plant Physiol 122: 403–413

    Article  PubMed  CAS  Google Scholar 

  • Fernandes KVS, Sabelli PA, Barratt DHP, Richardson M, Xavier-Filho J, Shewry PR (1993) The resistance of cowpea seeds to bruchid beetles is not related to levels of cysteine proteinase inhibitors. Plant Mol Biol 23: 215–219

    Article  PubMed  CAS  Google Scholar 

  • Gaddour K, Vicente-Carbajosa J, Lara P, Isabel-Lamoneda I, Diaz I, Carbonero P (2001) A constitutive cystatin-encoding gene from barley (Icy) responds differentially to abiotic stimuli. Plant Mol Biol 45: 599–608

    Article  PubMed  CAS  Google Scholar 

  • He X, Kermode AR (2003) Proteases associated with programmed cell death of megagametophyte cells after germination of white spruce (Piceaglauca) seeds. Plant Mol Biol 52: 729–744

    Article  PubMed  CAS  Google Scholar 

  • HongJK, Hwang JE, Lim CJ, Lee KO, Chung WS, Park B-S, Lim CO (2008) Molecular characterization of phytocystatins isolated from Chinese cabbage flower buds. Genes Genomics 30: 235–243

    Google Scholar 

  • Hong JK, Hwang JE, Lim CJ, Yang KA, Jin JL, Kim CY, Koo JC, Chung WS, Lee KO, Lee SY, Cho MJ, Lim CO (2007) Over-expression of Chinese cabbage phytocystatin 1 retards seed germination inArabidopsis. Plant Sci 172: 556–563

    Article  CAS  Google Scholar 

  • HongJK, Lee EY, Jin ZL, Yang KA, Choi YJ, Chung WS, Kim HI, Yun D-J, Lee SY, Cho MJ, Lim CO (2003) Expression of a Chinese cabbage cysteine proteinase inhibitor, BCPI-1, retards seed germination and early seedling growth rate. J Kor Soc Hort Sci 44: 553–559

    Google Scholar 

  • Irie K, Hosoyama H, Takeuch T, Iwabuchi K, Watanabe H, Abe M, Abe K, Arai S (1996) Transgenic rice established to express corn cystatin exhibits strong inhibitory activity against insect gut proteinases. Plant Mol Biol 30: 149–157

    Article  PubMed  CAS  Google Scholar 

  • Jin ZL, HongJK, Yang KA, Koo JC, Choi YJ, Chung WS, Yun D-J, Lee SY, Cho MJ, Lim CO (2005) Over-expression of Chinese cabbage calreticulin 1,BrCRT1, enhances shoot and root regeneration, but retards plant growth in transgenic tobacco. Transgenic Res 14: 619–626

    Article  PubMed  CAS  Google Scholar 

  • Jin ZL, HongJK, Yun D-J, Lee SY, Choi YJ, Bahk JD, Beachy RN, Cho MJ, Lim CO (2002) Expression of the tobacco mosaic virus movement protein alters starch accumulation inNicotiana benthamiana. J Plant Biol 45: 77–82

    Article  CAS  Google Scholar 

  • Katiyar-Agarwal S, Agarwal M, Grover A (2003) Heat-tolerant basmati rice engineered by over-expression of hsp101. Plant Mol Biol 51: 677–686

    Article  PubMed  CAS  Google Scholar 

  • Kondo H, Abe K, Emori Y, Arai S (1991) Gene organization of oryzacystatin-II, a new cystatin superfamily member of plant origin, is closely related to that of oryzacystatin-I but different from those of animal cystatins. FEBS Lett 278: 87–90

    Article  PubMed  CAS  Google Scholar 

  • Kondo H, Abe K, Nishimura I, Watanabe H, Emori Y, Arai S (1990) Two distinct cystatin species in rice seeds with different specificities against cysteine proteinases. Molecular cloning, expression, and biochemical studies on oryzacystatin-II. J Biol Chem 265: 15832–15837

    PubMed  CAS  Google Scholar 

  • Koo JC, Chun HJ, Park HC, Kim MC, Koo YD, Koo SC, Ok HM, Park SJ, Lee S-H, Yun D-J, Lim CO, Bahk JD, Lee SY, Cho MJ (2002) Over-expression of a seed specific hevein-like antimicrobial peptide fromPharbitis nil enhances resistance to a fungal pathogen in transgenic tobacco plants. Plant Mol Biol 50: 441–452

    Article  PubMed  CAS  Google Scholar 

  • Kuroda M, Kiyosaki T, Matsumoto I, Misaka T, Arai S, Abe K (2001) Molecular cloning, characterization, and expression of wheat cystatins. Biosci Biotechnol Biochem 65: 22–28

    Article  PubMed  CAS  Google Scholar 

  • Lecardonnel A, Chauvin L, Jouanin L, Beaujean A, Prevost G, Sang-wan-Norreel B (1999) Effects of rice cystatin I expression in transgenic potato on Colorado potato beetle larvae. Plant Sci 140: 71–79

    Article  CAS  Google Scholar 

  • Lim CO, Lee SI, Chung WS, Park SH, Hwang I, Cho MJ (1996) Characterization of a cDNA encoding cysteine proteinase inhibitor from Chinese cabbage (Brassica campestris L. ssp.pekinensis) flower buds. Plant Mol Biol 30: 373–379

    Article  PubMed  CAS  Google Scholar 

  • Margis R, Reis EM, Villeret V (1998) Structural and phylogenetic relationships among plant and animal cystatins. Arch Biochem Biophys 359: 24–30

    Article  PubMed  CAS  Google Scholar 

  • Martínez M, Rubio-Somoza I, Carbonero P, Diaz I (2003) A cathepsin B-like cysteine protease gene fromHordeum vulgare (geneCatB) induced by GA in aleurone cells is under circadian control in leaves. J Exp Bot 54: 951–959

    Article  PubMed  Google Scholar 

  • Misaka T, Kuroda M, Iwabuchi K, Abe K, Arai S (1996) Soyacystatin, a novel cysteine proteinase inhibitor in soybean, is distinct in protein structure and gene organization from other cystatins of animal and plant origin. Eur J Biochem 240: 609–614

    Article  PubMed  CAS  Google Scholar 

  • Masoud SA, Johnson LB, White FF, Reeck GR (1993) Expression of a cysteine proteinase inhibitor (oryzacystatin-I) in transgenic tobacco plants. Plant Mol Biol 21: 655–663

    Article  PubMed  CAS  Google Scholar 

  • Müntz K, Belozersky MJ, Dunaevsky YE, Schlerenth A, Tiedemann J (2001) Stored proteinases and the initiation of storage protein mobilization in seeds during germination and seedling growth. J Exp Bot 52: 1741–1752

    Article  PubMed  Google Scholar 

  • Pernas M, Sánchez-Monge R, Gómez L, Salcedo G (1998) A chestnut seed cystatin differentially effective against cysteine proteinases from closely related pests. Plant Mol Biol 38: 1235–1242

    Article  PubMed  CAS  Google Scholar 

  • Schlereth A, Standhardt D, Mock HP, Müntz K (2001) Stored cysteine proteinases start globulin mobilization in protein bodies of embryonic axes and cotyledons during vetch (Vicia sativa L.) seed germination. Planta 212: 718–727

    Article  PubMed  CAS  Google Scholar 

  • Solomon M, Belenghi B, Delledonne M, Menachem E, Levine A (1999) The involvement of cysteine proteases and protease inhibitor genes in the regulation of programmed cell death in plants. Plant Cell 11: 431–443

    Article  PubMed  CAS  Google Scholar 

  • Song I, Taylor M, Baker K, Bateman Jr. RC (1995) Inhibition of cysteine proteinases by Caricapapaya cystatin produced inEscherichia coli. Gene 162: 221–224

    Article  PubMed  CAS  Google Scholar 

  • Turk V, Bode W (1991) The cystatins: protein inhibitors of cysteine proteinases. FEBS 285: 213–219

    Article  CAS  Google Scholar 

  • Vain P, Worland B, Clarke MC, Richard G, Beavis M, Liu H, Kohli A, Leech M, Snape J, Christou P, Atkinson H (1998) Expression of an engineered cysteine proteinase inhibitor (Oryzacystatin-IDELTAD86) for nematode resistance in transgenic rice plants. Theor Appl Genet 96: 266–271

    Article  CAS  Google Scholar 

  • Zhao Y, Botella MA, Subramanian L, Niu X, Nielsen SS, Bressan RA, Hasegawa PM (1996) Two wound-inducible soybean cysteine proteinase inhibitors have greater insect digestive proteinase inhibitory activities than a constitutive homolog. Plant Physiol 111:1299–1306

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Environmental Biotechnology National Core Research Centre and PMBBRC, Division of Applied Life Science (BK21 Program), Graduate School of Gyeongsang National University, 660-701, Jinju, Republic of Korea

    Jung Eun Hwang, Woo Sik Chung, Kyun Oh Lee & Chae Oh Lim

  2. Brassica Genomics Team, National Institute of Agricultural Biotechnology, Rural Development Administration, 447-707, Suwon, Republic of Korea

    Joon Ki Hong & Beom -Seok Park

  3. Department of Food and Nutrition, Silla University, 617-736, Busan, Republic of Korea

    Young Ju Choi

  4. Department of Microbiological Engineering, Jinju National University, 660-758, Jinju, Republic of Korea

    Sang Wan Gal

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  1. Joon Ki Hong
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  2. Jung Eun Hwang
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  3. Woo Sik Chung
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  4. Kyun Oh Lee
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  5. Young Ju Choi
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  7. Beom -Seok Park
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  8. Chae Oh Lim
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Correspondence to Chae Oh Lim.

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Hong, J.K., Hwang, J.E., Chung, W.S. et al. Expression of a Chinese cabbage cysteine proteinase inhibitor, BrCYS1, retards seed germination and plant growth in transgenic Tobacco plant. J. Plant Biol. 51, 347–353 (2008). https://doi.org/10.1007/BF03036137

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  • DOI: https://doi.org/10.1007/BF03036137

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