Skip to main content
Log in

Assessment of Nematode Resistance in Wheat Transgenic Plants Expressing Potato Proteinase Inhibitor (PIN2) Gene

  • Published:
Transgenic Research Aims and scope Submit manuscript

Abstract

Serine proteinase inhibitors (IP’s) are proteins found naturally in a wide range of plants with a significant role in the natural defense system of plants against herbivores. The question addressed in the present study involves assessing the ability of the serine proteinase inhibitor in combating nematode infestation. The present study involves engineering a plant serine proteinase inhibitor (pin2) gene into T. durum PDW215 by Agrobacterium-mediated transformation to combat cereal cyst nematode (Heterodera avenae) infestation. Putative T0 transformants were screened and positive segregating lines analysed further for the study of the stable integration, expression and segregation of the genes. PCR, Southern analysis along with bar gene expression studies corroborate the stable integration pattern of the respective genes. The transformation efficiency is 3%, while the frequency of escapes was 35.71%. χ2 analysis reveals the stable integration and segregation of the genes in both the T1 and T2 progeny lines. The PIN2 systemic expression confers satisfactory nematode resistance. The correlation analysis suggests that at p < 0.05 level of significance the relative proteinase inhibitor (PI) values show a direct positive correlation vis-à-vis plant height, plant seed weight and also the seed number.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • HJ Atkinson (1993) Opportunities for improved control of plant parasitic nematodes via plant biotechnology DJ Beadle DHL Bishop LG Copping GK Dixon DW Holloman (Eds) Opportunities for Molecular Biology in Crop Production British Crop Protection Council Nottingham, UK 257–66

    Google Scholar 

  • HJ Atkinson P Urwin E Hansen M McPherson (1995) ArticleTitleDesigns for engineered resistance to root-parasitic nematodes Trends Biotechnol 13 369–374 Occurrence Handle10.1016/S0167-7799(00)88983-0

    Article  Google Scholar 

  • HJ Atkinson PE Urwin MJ McPherson (2003) ArticleTitleEngineering plants for nematode resistance Ann Rev Phytopathol 41 615–639 Occurrence Handle10.1146/annurev.phyto.41.052002.095737

    Article  Google Scholar 

  • M Cheng JE Fry S Pang H Zhou CM Hironaka DR Duncan TW Conner Y Wan (1997) ArticleTitleGenetic transformation of wheat mediated by Agrobacterium tumefaciens Plant Physiol 115 971–980 Occurrence Handle12223854

    PubMed  Google Scholar 

  • X Duan X Li Q Xue M Abo-El-Saad D Xu R Wu (1996) ArticleTitleTransgenic rice plants harboring an introduced potato proteinase inhibitor II gene are insect resistant Nature Biotechnol 14 494–498 Occurrence Handle10.1038/nbt0496-494

    Article  Google Scholar 

  • Hepher A and Atkinson HJ (1992) Nematode control with proteinase inhibitors. Patent WO92/15690.

  • T Hu S Metz C Chay HP Zhou N Biest G Chen M Cheng X Feng M Radionenko F Lu J Fry (2003) ArticleTitleAgrobacterium-mediated large-scale transformation of wheat (Triticum aestivum L.) using glyphosate selection Plant Cell Rep 21 1010–1019 Occurrence Handle10.1007/s00299-003-0617-6 Occurrence Handle12835912

    Article  PubMed  Google Scholar 

  • RS Hussey (1993) ArticleTitleEvaluation of transgenic plants expressing proteinase inhibitor genes for root-knot nematode Soc Nematol Mol Biol Newslett 5 15–16

    Google Scholar 

  • RS Hussey FMW Grundler (1998) Nematode parasitism of plants RN Perry DJ Wright (Eds) The Physiology and Biochemistry of Free- Living and Plant-Parasitic Nematodes CABI Publishing Wallingford 43–213

    Google Scholar 

  • RA Jefferson TA Kavanagh MW Bevan (1987) ArticleTitleGUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants EMBO J 6 3901–3907 Occurrence Handle3327686

    PubMed  Google Scholar 

  • R Johnson J Narvaez G An C Ryan (1989) ArticleTitleExpression of proteinase inhibitors I and II in transgenic tobacco plants: effects on natural defense against Manduca sexta larvae Proc Natl Acad Sci (USA) 86 9871–9875

    Google Scholar 

  • HK Khanna GE Daggard (2003) ArticleTitleAgrobacterium tumefaciens-mediated transformation of wheat using a superbinary vector and a polyamine-supplemented regeneration medium Plant Cell Rep 21 429–36 Occurrence Handle12789445

    PubMed  Google Scholar 

  • J Khurana A Chugh P Khurana (2002) ArticleTitleRegeneration from mature and immature embryos and transient gene expression via Agrobacterium-mediated transformation in emmer wheat (Triticum dicoccum Schuble) Indian J Exp Biol 40 1295–303 Occurrence Handle13677634

    PubMed  Google Scholar 

  • C Kramer J Di Mio GK Carswell RD Shillito (1993) ArticleTitleSelection of transformed protoplast-derived Zea mays colonies with phosphinothricin and a novel assay using the pH indicator chlorophenol red Planta 190 454–458 Occurrence Handle10.1007/BF00224783

    Article  Google Scholar 

  • Lawrence PK and Koundal KR (2002) Plant protease inhibitors in control of phytophagous insects. Electronic J. Biotechnol 5: 93–104. (http://www.ejb.org/content/vol5/issue1).

    Google Scholar 

  • B Lee K Murdoch M Kreis MGK Jones (1989) ArticleTitleA method for large-scale progeny screening of putative transformed cereal Plant Mol Biol Rep 7 129–134

    Google Scholar 

  • CJ Lilley PE Urwin HJ Atkinson MJ McPherson (1997) ArticleTitleCharacterization of cDNAs encoding serine proteinases from the soybean cyst nematode Heterodera glycines Mol Biochem Parasitol 89 195–207 Occurrence Handle10.1016/S0166-6851(97)00116-3 Occurrence Handle9364965

    Article  PubMed  Google Scholar 

  • CJ Lilley PE Urwin MJ McPherson HJ Atkinson (1996) ArticleTitleCharacterisation of intestinally active proteinases of cyst nematodes Parasitol 113 415–425

    Google Scholar 

  • DM Lonsdale S Lindup LJ Moisan AJ Harvey (1998) ArticleTitleUsing firefly luciferase to identify the transition from transient to stable expression in bombarded wheat scutellar tissue Physiol Plant 102 447–453 Occurrence Handle10.1034/j.1399-3054.1998.1020313.x

    Article  Google Scholar 

  • A Mahalakshmi A Chugh P Khurana (2000) ArticleTitleExogenous DNA uptake via cellular perneabilization and expression of foreign gene in wheat zygotic embryos Plant Biotech 17 235–240

    Google Scholar 

  • MT McManus EPJ Burgess B Philip LM Watson WA Laing CR Voisey DWR White (1999) ArticleTitleExpression of the soybean (Kunitz) trypsin inhibitor in transgenic tobacco: Effects on larval development of Spodoptera litura Transgenic Res 8 383–395 Occurrence Handle10.1023/A:1008957610872

    Article  Google Scholar 

  • KB Mullis F Faloona (1987) ArticleTitleSpecific synthesis of DNA in vitro via a polymerase chain reaction Method Enzymol 155 350–355

    Google Scholar 

  • NS Nehra RN Chibbar N Leung K Caswell C Mallard L Steinhauer M Baga KK Kartha (1994) ArticleTitleSelf-fertile transgenic wheat plants regenerated from isolated scutellar tissues from microprojectile bombardment with two distinct gene constructs Plant J 5 285–297 Occurrence Handle10.1046/j.1365-313X.1994.05020285.x

    Article  Google Scholar 

  • M Ozgen M Turet S Ozcan C Sancak (1996a) ArticleTitleCallus induction and plant regeneration from immature and mature embryos of winter durum wheat genotypes Plant Breed 115 455–458

    Google Scholar 

  • M Ozgen M Turet S Altinok C Sanzak (1996b) ArticleTitleEfficient callus induction and plant regeneration from mature embryo culture of winter wheat (Triticum aestivum L.) genotypes Plant Cell Rep 18 331–335

    Google Scholar 

  • Patnaik D, Khurana P (2001) Wheat Biotechnology: a minireview. Electronic J Biotechnol 4: 1–29. (http://www.ejb.org/content/vol4/issue2/full/4/).

    Google Scholar 

  • D Patnaik Khurana P (2003) ArticleTitleGenetic transformation of Indian bread (T. aestivum) and pasta (T. durum) wheat by particle bombardment of mature embryo-derived calli. BMC Plant Biology 3 5–16 Occurrence Handle10.1186/1471-2229-3-5 Occurrence Handle12952555

    Article  PubMed  Google Scholar 

  • Patnaik D and Vishnudasan D, Khurana P (2005) Agrobacterium-mediated transformation of mature embryos of Triticum aestivum and Triticum durum. BMC Biotechnol (in press).

  • A Pellegrineschi R Brito L Velazquez L Noguera W Pfeiffer S McLean D Hoisington (2002) ArticleTitleThe effect of pretreatment with mild heat and drought stresses on the explant and biolistic transformation frequency of three durum wheat cultivars Plant Cell Rep 20 955–960 Occurrence Handle10.1007/s00299-002-0438-z

    Article  Google Scholar 

  • CA Ryan (1990) ArticleTitleProtease inhibitors in plants: genes for improving defenses against insects and pathogens Annu Rev Phytopathol 28 425–449 Occurrence Handle10.1146/annurev.py.28.090190.002233

    Article  Google Scholar 

  • Sasser JN, Freckman DW (1987) A world perspective on nematology: The role of the society. In: Veech JA, Dickerson DW (eds), Vistas on nematology. (pp. 7–14) Society of Nematologists.

  • V Srivastava OD Anderson DW Ow (1999) ArticleTitleSingle-copy transgenic wheat generated through the resolution of complex integration patterns Proc Natl Acad Sci (USA) 96 11117–11121 Occurrence Handle10.1073/pnas.96.20.11117

    Article  Google Scholar 

  • S Takumi T Shimada (1996) ArticleTitleProduction of transgenic wheat through particle bombardment of scutellar tissues: frequency is influenced by culture duration J Plant Physiol 149 418–423

    Google Scholar 

  • PE Urwin CJ Lilley MJ McPherson HJ Atkinson (1997) ArticleTitleResistance to both cyst and root-knot nematodes conferred by transgenic Arabidopsis expressing a modified plant cystatin Plant J 12 455–461 Occurrence Handle10.1046/j.1365-313X.1997.12020455.x Occurrence Handle9301094

    Article  PubMed  Google Scholar 

  • D Vishnudasan P Khurana (2005) ArticleTitleNew paradigms towards appraising plant parasitic nematodes infestation with special emphasis on Cereal Cyst Nematode (H.avenae) Physiol Mol Biol plants 11 1–18

    Google Scholar 

  • H Wu C Sparks B Amoah HD Jones (2003) ArticleTitleFactors influencing successful Agrobacterium-mediated genetic transformation of wheat Plant Cell Rep 21 659–68 Occurrence Handle12789416

    PubMed  Google Scholar 

  • U Wyss U Zunke (1986) ArticleTitleObservations on the behaviour of second stage juveniles of Heterodera schachtii inside host roots Rev Nematol 9 153–165

    Google Scholar 

  • D Xu D McElroy RW Thornburg R Wu (1993) ArticleTitleSystemic induction of a potato pin2 promoter by wounding, methyl jasmonate and abscisic acid in transgenic rice plants Plant Mol Biol 22 573–588 Occurrence Handle10.1007/BF00047399 Occurrence Handle8343596

    Article  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Vishnudasan, D., Tripathi, M., Rao, U. et al. Assessment of Nematode Resistance in Wheat Transgenic Plants Expressing Potato Proteinase Inhibitor (PIN2) Gene. Transgenic Res 14, 665–675 (2005). https://doi.org/10.1007/s11248-005-5696-4

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11248-005-5696-4

Keywords

Navigation