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Structural diversity and organization of three gene families for Kunitz-type enzyme inhibitors from potato tubers ( Solanum tuberosum L.)

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Abstract

In the potato, Kunitz-type enzyme inhibitors are abundant and highly polymorphic small proteins found in tubers. DNA sequence analysis of 1596 unselected ESTs (expressed sequence tags) from mature tubers of the cultivars Provita and Saturna resulted in the identification of 55 different DNA sequences with high sequence similarity to Kunitz-type enzyme inhibitors. The frequency of Kunitz-type inhibitor ESTs in Provita was four times higher than in Saturna tubers, and none of the Provita ESTs was identical to any of the Saturna ESTs. A phenogram constructed from the deduced amino acid sequences of the inhibitors revealed three major homology groups—A, B and C. Group A inhibitors were all derived from Provita ESTs. Inhibitor groups A and B were more similar to each other than to group C inhibitors, and for most members within-group similarity was at least 90%. Non-conservative amino acid substitutions and insertion/deletion polymorphisms suggest functional differentiation between members of the gene family. A minimum of 21 genes for Kunitz-type enzyme inhibitors (six for group A, nine for group B and six for group C) was estimated to exist in the potato genome. Genetic mapping and the identification of BAC (bacterial artificial chromosome) clones containing more than one member of the gene family indicated that most inhibitor genes of groups A, B and C are organized in a cluster that maps to a single region on potato chromosome III.

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References

  • Ballvora A, Ercolano MR, Weiß J, Meksem K, Bormann CA, Oberhagemann P, Salamini F, Gebhardt C (2002) The R1 gene for potato resistance to late blight ( Phytophthora infestans) belongs to the leucine zipper/NBS/LRR class of plant resistance genes. Plant J 30:361–371

    Article  PubMed  Google Scholar 

  • Bánfalvi Z, Molnár A, Molnár G, Lakatos L, Szabó L (1996) Starch synthesis, and tuber storage protein genes are differently expressed in Solanum tuberosum and in Solanum brevidens. FEBS Lett 383, 159–164

    Google Scholar 

  • Bishop JG, Dean AM, Mitchell-Olds T (2000) Rapid evolution in plant chitinases: molecular targets of selection in plant-pathogen coevolution. Proc Natl Acad Sci USA 97:5322–5327

    Article  CAS  PubMed  Google Scholar 

  • Bohac JR (1991) A modified method to purify patatin from potato tubers. J Agric Food Chem 39:1411–1415

    CAS  Google Scholar 

  • Bonierbale MW, Plaisted RL, Tanksley SD (1988) RFLP maps based on a common set of clones reveal modes of chromosomal evolution in potato and tomato. Genetics 120:1095–1103

    Google Scholar 

  • Boulter D (1993) Insect pest control by copying nature using genetically engineered crops. Phytochemistry 34:1453–1466

    Article  CAS  PubMed  Google Scholar 

  • Devereux J, Haeberli P, Smithies O (1984) A comprehensive set of sequence analysis programmes for the VAX. Nucl Acids Res 12:387–395

    CAS  Google Scholar 

  • Gebhardt C, Ritter E, Debener T, Schachtschabel U, Walkemeier B, Uhrig H, Salamini F (1989a) RFLP analysis and linkage mapping in Solanum tuberosum. Theor Appl Genet 78:65–75

    Google Scholar 

  • Gebhardt C, Blomendahl C, Schachtschabel U, Debener T, Salamini F, Ritter E (1989b) Identification of 2n breeding lines and 4n varieties of potato ( Solanum tuberosum, ssp tuberosum) with RFLP-fingerprints. Theor Appl Genet 78:16–22

    Google Scholar 

  • Gebhardt C, Ritter E, Barone A, Debener T, Walkemeier B, Schachtschabel U, Kaufmann H, Thompson RD, Bonierbale MW, Ganal MW, Tanksley SD, Salamini F (1991) RFLP maps of potato and their alignment with the homeologous tomato genome. Theor Appl Genet 83:49–57

    Google Scholar 

  • Glaczinski H, Heibges A, Salamini F, Gebhardt C (2003) The proteinaceous inhibitor of soluble potato tuber invertase is a member of a gene family known as Kunitz-type proteinase inhibitors. Potato Res, in press

  • Gruden C, Strukelj B, Ravnikar M, Poljsak-Prijatelj M, Mavric I, Brzin J, Pungercar J, Kregar I (1997) Potato cysteine proteinase inhibitor gene family: molecular cloning, characterization and immunocytochemical localization studies. Plant Mol Biol 34:317–323

    Article  CAS  PubMed  Google Scholar 

  • Hannapel DJ (1994) Nucleotide and deduced amino acid sequence of the 22-kilodalton cathepsin D inhibitor of potato ( Solanum tuberosum L.) Plant Physiol 101:703–704

  • Heibges A, Salamini F, Gebhardt C (2003) Functional comparison of homologous members of three groups of Kunitz-type enzyme inhibitors from potato tubers ( Solanum tuberosum L.). Mol Genet Genomics (following paper)

  • Hoffman LM, Sengupta-Gopalanm C, Paaren HE (1984) Structure of soybean Kunitz trypsin inhibitor mRNA determined from cDNA by using oligodeoxynucleotide primers. Plant Mol Biol 3:111–117

    CAS  Google Scholar 

  • Ishikawa A, Ohta S, Matsuoka K, Hattori T, Nakamura K (1994) A family of potato genes that encode Kunitz-type proteinase inhibitors: structural comparisons and differential expression. Plant Cell Physiol 35:303–312

    CAS  PubMed  Google Scholar 

  • Koide T, Ikenaka T (1973) Studies on soybean trypsin inhibitors. 3. Amino acid sequences of the carboxyl-terminal region and the complete amino acid sequence of soybean trypsin inhibitor (Kunitz). Eur J Biochem 32:417–431

    CAS  PubMed  Google Scholar 

  • Kortt AA, Strike PM, De Jersey J (1989) Amino acid sequence of a crystalline seed albumin (winged bean albumin-1) from Psophocarpus tetragonolobus (L.) DC. Sequence similarity with Kunitz-type seed inhibitors and 7S storage globulins. European J Biochem 181:403–408

    CAS  Google Scholar 

  • Kunitz M (1945) Crystallization of a trypsin inhibitor from soybean. Science 101:668–669

    CAS  Google Scholar 

  • Laskowski M Jr, Kato I, Kohr WJ, Park SJ, Tashiro M, Whatley HE (1987) Positive Darwinian selection in evolution of protein inhibitors of serine proteinase. Cold Spring Harbor Symp Quant Biol 52:545–553

    CAS  PubMed  Google Scholar 

  • Leister D, Ballvora A, Salamini F, Gebhardt C (1996) A PCR based approach for isolating pathogen resistance genes from potato with potential for wide application in plants. Nature Genet 14:421–429

    CAS  PubMed  Google Scholar 

  • Logemann J, Schell J, Willmitzer L (1987) Improved method for the isolation of RNA from plant tissues. Anal Biochem 163:16–20

    CAS  PubMed  Google Scholar 

  • Maniatis T, Frisch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Habour Laboratory Press, NY

    Google Scholar 

  • Mares R, Meloun B, Pavlik M, Kostka V, Baudys M (1989) Primary structure of cathepsin D inhibitor from potatoes and its structure relationship to soybean trypsin inhibitor family. FEBS Lett 251:94–98

    Article  CAS  PubMed  Google Scholar 

  • Matsuoka K, Nakamura K (1991) Propeptide of a precursor to a plant vacuolar protein required for vacuolar targeting. Proc Natl Acad Sci USA 88:834–838

    CAS  PubMed  Google Scholar 

  • Matsuoka K, Matsuomo S, Hattori T, Machida Y, Nakamura K (1990) Vacuolar targeting and post-translational processing of the precursor to the sweet potato tuberous root storage protein in heterologous plant cells. J Biol Chem 265:19750–19757

    CAS  PubMed  Google Scholar 

  • Nakamura K, Matsuoka K (1993) Protein targeting to the vacuole in plant cells. Plant Physiol 101:1-5

    Article  CAS  PubMed  Google Scholar 

  • Park KS, Cheong JJ, Lee SJ, Suh MC, Choi D (2000) A novel proteinase inhibitor gene transiently induced by tobacco mosaic virus infection. Biochim Biophys Acta 1492:509–512

    Article  CAS  PubMed  Google Scholar 

  • Peňa-Cortes H, Sanchez-Serrano J, Rocha-Sosa M, Willmitzer L (1988) Systemic induction of proteinase inhibitor II gene expression in potato plants by wounding. Planta 174:84–89

    Google Scholar 

  • Ritonja A, Krizaj I, Mesko P, Kopitar M, Lucovnik P, Strukelj B, Pungercar J, Buttle DJ, Barret AJ, Turk V (1990) The amino acid sequence of a novel inhibitor of cathepsin D from potato. FEBS Lett 267:13–15

    Article  CAS  PubMed  Google Scholar 

  • Ryan CA (1990) Protease inhibitors in plants: genes for improving defenses against insects and pathogens. Annu Rev Phytopathology 28:425–449

    Article  CAS  Google Scholar 

  • Schuler TH, Poppy GM, Kerry BR, Denholm I (1998) Insect-resistant transgenic plants. Trends Biotech 16:168–175

    Article  CAS  Google Scholar 

  • Shewry PR (1999) Enzyme inhibitors of seeds: types and properties. In: Shewry PR, Casey R (eds) Seed proteins. Kluwer Academic Publishers, Amsterdam, pp 587–615

  • Stegemann H, Loeschke V (1976) Index of European potato varieties. Mitteilungen aus der Biologischen Bundesanstalt für Land- und Forstwirtschaft, Berlin-Dahlem, Heft 168

  • Stiekema WJ, Heidekamp F, Dirkse WG, Beckum JV, Haan PD, Bosch CT, Louserse JD (1988) Molecular cloning and analysis of four potato tuber mRNAs. Plant Mol Biol 11:255–269

    CAS  Google Scholar 

  • Strickland JA, Orr GL, Walsh TA (1995) Inhibition of larval growth by patatin, the lipid acylhydrolase from potato tubers. Plant Physiol 109:667–674

    CAS  PubMed  Google Scholar 

  • Strukelj B, Pungercar J, Ritonja A, Krizaj I, Gubensek F, Kregar I, Turk V (1990) Nucleotide and deduced amino acid sequence of an aspartic proteinase inhibitor homologue from potato tubers ( Solanum tuberosum L.) Nucleic Acids Res 18:4605

  • Strukelj B, Ravnikar M, Mesko P, Poljsak-Prijatelj M, Pungercar J, Kopitar G, Kregar I. Turk V (1995) Molecular cloning and immunocytochemical localization of jasmonic acid inducible cathepsin D inhibitor from potato. Adv Exp Med Biol 362:293–298

    CAS  PubMed  Google Scholar 

  • Suh SG, Peterson JE, Stiekema WJ, Hannapel DJ (1990) Purification and characterization of the 22-kilodalton potato tuber proteins. Plant Physiol 94:40–45

    CAS  Google Scholar 

  • Svendsen IB, Hejgaard J, Mundy J (1986) Complete amino acid sequence of the α-amylase/subtilisin inhibitor from barley. Carlsberg Res Comm 51:43–50

    CAS  Google Scholar 

  • Thornburg RW, An G, Cleveland TE, Johnson R, Ryan CA (1987) Wound-inducible expression of a potato inhibitor II-chloramphenicol acetyl-transferase gene fusion in transgenic tobacco plants. Proc Natl Acad Sci USA 84:744–748

    CAS  Google Scholar 

  • Tiffin B, Gaut BS (2001) Molecular evolution of the wound-induced protease inhibitor wip1 in Zea and related genera. Mol Biol Evol 18:2091–2101

    Google Scholar 

  • Walsh TA, Twitchell WP (1991) Two Kunitz-type proteinase inhibitors from potato tubers. Plant Physiol 97:15–18

    CAS  Google Scholar 

  • Weselake RJ, MacGregor AW, Hill RD (1985) Endogenous alpha-amylase inhibitor in various cereals. Cereal Chem 62:120–123

    CAS  Google Scholar 

  • Yamagishi K, Mitsumori C, Kikuta Y (1991) Nucleotide sequence of a cDNA encoding the putative trypsin inhibitor in potato tuber. Plant Mol Biol 17:287–288

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

Part of this work was supported by the European Union under ECLAIR-ERL Project No. 0034

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Authors and Affiliations

  1. Max-Planck-Institut für Züchtungsforschung, Carl-von-Linné-Weg 10, 50829, Köln, Germany

    A. Heibges, H. Glaczinski, A. Ballvora, F. Salamini & C. Gebhardt

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  1. A. Heibges
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  2. H. Glaczinski
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  3. A. Ballvora
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  4. F. Salamini
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  5. C. Gebhardt
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Correspondence to C. Gebhardt.

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Communicated by R. G. Herrmann

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Heibges, A., Glaczinski, H., Ballvora, A. et al. Structural diversity and organization of three gene families for Kunitz-type enzyme inhibitors from potato tubers ( Solanum tuberosum L.). Mol Gen Genomics 269, 526–534 (2003). https://doi.org/10.1007/s00438-003-0860-0

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