Advertisement

Journal of Biosciences

, Volume 30, Issue 5, pp 627–638 | Cite as

Enhancement of resistance to aphids by introducing the snowdrop lectin genegna into maize plants

  • Zhaoyu Wang
  • Kewei Zhang
  • Xiaofen Sun
  • Kexuan Tang
  • Juren Zhang
Article

Abstract

In order to enhance the resistance to pests, transgenic maize (Zea mays L.) plants from elite inbred lines containing the gene encoding snowdrop lectin (Galanthus nivalis L. agglutinin; GNA) under control of a phloem-specific promoter were generated through theAgrobacterium tumefaciens- mediated method. The toxicity of GNA-expressing plants to aphids has also been studied. The independently derived plants were subjected to molecular analyses. Polymerase chain reaction (PCR) and Southern blot analyses confirmed that thegna gene was integrated into maize genome and inherited to the following generations. The typical Mendelian patterns of inheritance occurred in most cases. The level of GNA expression at 0.13%-0.28% of total soluble protein was observed in different transgenic plants. The progeny of nine GNA-expressing independent transformants that were derived separately from the elite inbred lines DH4866, DH9942, and 8902, were selected for examination of resistance to aphids. These plants synthesized GNA at levels above 0.22% total soluble protein, and enhanced resistance to aphids was demonstrated by exposing the plants to corn leaf aphid (Rhopalosiphum maidis Fitch) under greenhouse conditions. The nymph production was significantly reduced by 46.9% on GNA-expressing plants. Field evaluation of the transgenic plants supported the results from the inoculation trial. After a series of artificial self-crosses, some homozygous transgenic maize lines expressing GNA were obtained. In the present study, we have obtained new insect-resistant maize material for further breeding work.

Keywords

Agrobacterium tumefaciens aphid Galanthus nivalis L. agglutinin gene maize transgene 

Abbreviations used

AP

Alkaline phosphatase

DI

damage index

GNA

Galanthus nivalis L. agglutinin

PCR

polymerase chain reaction

RDI

relative damage index

RSs-1

rice sucrose synthase-1

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Ausubel F M, Brent R, Kingston R E, Moore D D, Seidman J G, Smith J A and Struhl K 1995a Preparation of genomic DNA from plant tissue; inShort protocols in molecular biology 3rd edition (sed.) E J Richards (Beijing: Science Press) pp 36–39Google Scholar
  2. Ausubel F M, Brent R, Kingston R E, Moore D D, Seidman J G, Smith J A and Struhl K 1995b Southern blotting; inShort protocols in molecular biology 3rd edition (sed.) T Brown (Beijing: Science Press) pp 55–60Google Scholar
  3. Ausubel F M, Brent R, Kingston R E, Moore D D, Seidman J G, Smith J A and Struhl K 1995c Immunology; inShort protocols in molecular biology 3rd edition (seds) H M Cooper and Y Paterson (Beijing: Science Press) pp 411–441Google Scholar
  4. Ausubel F M, Brent R, Kingston R E, Moore D D, Seidman J G, Smith J A and Struhl K 1995d Analysis of proteins; inShort protocols in molecular biology 3rd edition (seds) J A Smith, S R Gallagher and S E Winston (Beijing: Science Press) pp 329–373Google Scholar
  5. Cristofoletti P T, Ribeiro A F, Deraison C, Rahb Y and Terra W R 2003 Midgut adaptation and digestive enzyme distribution in a phloem feeding insect, the pea aphidAcyrthosiphon pisum;J. Insect Physiol. 49 11–24PubMedCrossRefGoogle Scholar
  6. Deraison C, Darboux I, Duportets L, Gorojankina T, Rahb Y and Jouanin L 2004 Cloning and characterization of a gut-specific cathepsin L from the aphidAphis gossypii;Insect Mol. Biol. 13 165–177PubMedCrossRefGoogle Scholar
  7. Down R E, Gatehouse AMR, Hamilton W D O and Gatehouse J A 1996 Snowdrop lectin inhibits development and fecundity of the glasshouse potato aphid (Aulacorthum solani) when administeredin vitro and via transgenic plants, both in laboratory and glasshouse trials;J. Insect Physiol. 42 pp 1035–1045CrossRefGoogle Scholar
  8. Eisemann C H, Donaldson R A, Pearson R D, Cadagon L C, Vuocolo T and Pellam R L 1994 Larvicidal action of lectins onLucilia cuprina; mechanism of action;Entomol. Exp. Appl. 72 1–11CrossRefGoogle Scholar
  9. Escudero J, Neuhaus G, Schlappi M and Hohn B 1996 T-DNA transfer in meristematic cells of maize provided with intracellularAgrobacterium;Plant J. 10 355–360CrossRefGoogle Scholar
  10. Frame B R, Shou H, Chikwamba R K, Zhang Z, Xiang C, Fonger T M, Sue Ellen K P, Li B, Nettleton D S, Pei D and Wang K 2002Agrobacterium tumefaciens-mediated transformation of maize embryos using a standard binary vector system;Plant Physiol. 129 13–22PubMedCrossRefGoogle Scholar
  11. Foissac X, Edwards M G, Du J P, Gatehouse AMR and Gatehouse J A 2002 Putative protein digestion in a sapsucking homopteran plant pest (rice brown planthopper;Nilaparvata lugens; Delphacidae) — identification of trypsin-like and cathepsin B-like proteases;Insect Biochem. Mol. 39 967–978CrossRefGoogle Scholar
  12. Gatehouse AMR, Down R E, Powell K S, Sauvion N, Rahb Y, Newell C A, Merryweather A, Hamilton W D O and Gatehouse J A 1996 Transgenic potato plants with enhanced resistance to the peach-potato aphidMyzus persicae;Entomol. Exp. Appl. 79 295–307CrossRefGoogle Scholar
  13. Habibi J, Backus E A and Czalpa T H 1993 Plant lectins affect survival of the potato leafhopper (Homoptera: Cicadellidae);J. Econ. Entomol. 86 945–951Google Scholar
  14. Hilder V A, Gatehouse AMR, Sheerman S E, Barker R F and Boulter D 1987 A novel mechanism of insect resistance engineered into tobacco;Nature (London) 330 160–163CrossRefGoogle Scholar
  15. Hilder V A, Powell K S, Gatehouse AMR, Gatehouse J A, Shi Y, Hamilton W D O, Merryweather A, Newell C A, Timans J C, Peumans W J, Van Damme E and Boulter D 1995 Expression of snowdrop lectin in transgenic tobacco plants results in added protection against aphids;Transgenic Res. 4 18–25CrossRefGoogle Scholar
  16. Hu Q A 2000Agrobacterium-mediated transformation of rice hybrid parental lines using snowdrop lectin gene (gna); Doctoral dissertation, Fudan University, Shanghai, ChinaGoogle Scholar
  17. Ishida Y, Saito H, Ohta S, Hiei Y, Komari T and Kumashiro T 1996 High efficiency transformation of maize (Zea mays L.) mediated byAgrobacterium tumefaciens;Nat. Biotechnol. 14 745–750PubMedCrossRefGoogle Scholar
  18. Iyer L M, Kumpatla S P, Chandrasekharan M B and Hall T C 2000 Transgene silencing in monocots;Plant Mol. Biol. 43 323–346PubMedCrossRefGoogle Scholar
  19. Kumar S and Fladung M 2000 Transgene repeats in aspen: molecular characterisation suggests simultaneous integration of independent T-DNAs into receptive hotspots in the host genome;Mol. Gen. Genet. 264 20–28PubMedCrossRefGoogle Scholar
  20. Li G S, Yang A F, Zhang J R, Bi Y P and Shan L 2001 Genetic transformation of calli from maize and regeneration of herbicide-resistant plantlets;Chin. Sci. Bull. 46 563–565CrossRefGoogle Scholar
  21. Matzke M A, Scheid O M and Matzke A J 1999 Rapid structural and epigenetic changes in polyploid and aneuploid genomes;Bioessays 21 761–767PubMedCrossRefGoogle Scholar
  22. Powell K S, Gatehouse AMR, Hilder V A and Gatehouse J A 1993 Antimetabolic effects of plant lectins and plant and fungal enzymes on the nymphal stage of two important rice pests,Nilaparvata lugens andNephotettix nigropictuc;Entomol. Exp. Appl. 66 119–126CrossRefGoogle Scholar
  23. Powell K S, Gatehouse AMR, Hilder V A, van Damme E J, Peumans W J, Boonjawat J, Horsham K and Gatehouse J A 1995 Different antimetabolic effects of related lectins towards nymphal stages ofNilaparvata lugens;Entomol. Exp. Appl. 75 51–59CrossRefGoogle Scholar
  24. Powell K S, Spence J, Bharathi M, Gatehouse J and Gatehouse AMR 1998 Immunohistochemical and developmental studies to elucidate the mechanism of action of the snowdrop lectin on the rice brown planthopper,Nilaparvata lugens (Stl.);J. Insect Physiol. 44 529–539PubMedCrossRefGoogle Scholar
  25. Quan R D, Shang M, Zhang H, Zhao Y X and Zhang J R 2004 Improved chilling tolerance by transformation withbetA gene for the enhancement of glycinebetaine synthesis in maize;Plant Sci. 166 141–149CrossRefGoogle Scholar
  26. Rahb Y and Febvay G 1993 Protein toxicity to aphids: anin vitro test onAcyrthosiphon pisum;Entomol. Exp. Appl. 67 149–160CrossRefGoogle Scholar
  27. Rao K V, Rathore K S, Hodges T K, Fu X, Stoger E, Sudhakar, D Williams S, Christou P, Bharathi M, Bown D P, Powell K S, Spence J, Gatehouse A M R and Gatehouse J A 1998 Expression of snowdrop lectin (GNA) in transgenic rice plants confers resistance to rice brown planthopper;Plant J. 15 469–477PubMedCrossRefGoogle Scholar
  28. Sauvion N, Rahb Y, Peumans W J, VanDamme E J M, Gate-house J A and Gatehouse AMR 1996 Effects of GNA and other mannose-binding lectins on development and fecundity of the peach-potato aphid;Entomol. Exp. Appl. 79 285–293CrossRefGoogle Scholar
  29. Schlappi M and Hohn B 1992 Competence of immature maize embryos forAgrobacterium-mediated gene transfer;Plant Cell 4 7–16PubMedCrossRefGoogle Scholar
  30. Sharma H C and Ortiz R 2000 Transgenics, pest management, and the environment;Curr. Sci. 79 421–437Google Scholar
  31. Shen W H, Escudero J, Schlappi M, Ramos C, Hohn B and Koukolikova-Nicola Z 1993 T-DNA transfer to maize cells: histochemical investigation of beta-glucuronidase activity in maize tissues;Proc. Natl. Acad. Sci. USA 90 1488–1492PubMedCrossRefGoogle Scholar
  32. Shi Y, Wang M B, Powell K S, van Damme E, Hilder V A, Gatehouse AMR, Boulter D and Gatehouse J A 1994 Use of the rice sucrose synthase-1 promoter to direct phloem specific expression of Β-glucuronidase and snowdrop lectin genes in transgenic tobacco plants;J. Exp. Bot. 45 623–631CrossRefGoogle Scholar
  33. Stoger E, Williams S, Christou P, Down R E and Gatehouse J A 1999 Expression of the insecticidal lectin from snowdrop (Galanthus nivalis agglutinin; GNA) in transgenic wheat plants: effects on predation by the grain aphidSitobion avenae;Mol. Breed. 5 65–73CrossRefGoogle Scholar
  34. Sudhakar D, Fu X, Stoger E, Williams S, Spence J, Brown D P, Bharathi M, Gatehouse J A and Christou P 1998 Expression and immunolocalisation of the snowdrop lectin, GNA in transgenic rice plants;Transgenic Res. 7 371–378PubMedCrossRefGoogle Scholar
  35. Sun X F, Tang K X, Wan B L, Qi H X and Lu X G 2001 Transgenic rice homozygous lines expressing GNA showed enhanced resistance to rice brown planthopper;Chin. Sci. Bull. 46 1108–1113Google Scholar
  36. Tang K X, Tinjuangjun P, Xu Y N, Sun X F, Gatehouse J A, Ronald P C, Qi H X, Lu X G, Christou P and Kohli A 1999 Particle-bombardment-mediated co-transformation of elite Chinese rice cultivars with genes conferring resistance to bacterial blight and sap-sucking insect pests;Planta 208 552–563CrossRefGoogle Scholar
  37. Teng H T, Zhao J R, Guo J L, Wang Y D, Xing J F and Duan M X 2002 Report on transgenic maize breeding with borer-resistance;J. Maize Sci. 10 14–16Google Scholar
  38. Wang Z B and Guo S D 1999 Transgenic tobacco co-expressingcryIA andgna resisted both of the cotton boll worm and aphids;Chin. Sci. Bull. 44 2068–2075Google Scholar
  39. Wang Z Y, Sun X F, Wang F, Tang K X and Zhang J R 2005 Enhanced Resistance of Snowdrop Lectin (Galanthus nivalis L. agglutinin)-Expressing Maize (Zea mays L.) to Asian Corn Borer (Ostrinia furnacalis Guenée);J. Integrative Plant Biol. 47 873–880CrossRefGoogle Scholar
  40. WhittenM J and Oakenshott J G 1991 Opportunities for modern biotechnology in control of insect pests and weeds with special reference to developing countries;FAO Plant Prot. Bull. 39 155–181Google Scholar
  41. Wu C Y, Ye Z B, Li H X and Tang K X 2000 Genetic transformation of tomato with snowdrop lectin gene (GNA);Acta Bot. Sin. 42 719–723Google Scholar
  42. Xiao S H, Liu J G and Huang J Q 2001 Breeding of insecticidal transgenic cotton cultivations;Chin. Cotton 28 19–20Google Scholar
  43. Zhang G X and Zhong T S 1983Economic insect fauna of China 1st edition, Vol. 25 (Beijing: Science Press) pp 1–65Google Scholar
  44. Zhang X G 1992The research on the resistance of soybean cultivars to Cercospora sojina Hara, Doctoral dissertation, North-East Agricultural University, Harbin, Heilongjiang, ChinaGoogle Scholar
  45. Zhao Z Y, Cai T, Tagliani L, Wang N, Pang H, Rudert M, Schroeder S, Hondred D, Seltzer J and Pierce D 2000Agrobacterium-mediated sorghum transformation;Plant Mol. Biol. 44 789–798PubMedCrossRefGoogle Scholar

Copyright information

© Indian Academy of Sciences 2005

Authors and Affiliations

  • Zhaoyu Wang
    • 1
  • Kewei Zhang
    • 1
  • Xiaofen Sun
    • 2
  • Kexuan Tang
    • 2
    • 3
  • Juren Zhang
    • 1
  1. 1.Life Science SchoolShandong UniversityJinanPeople’s Republic of China
  2. 2.State Key Laboratory of Genetic EngineeringSchool of Life Sciences, Fudan UniversityShanghaiPeople’s Republic of China
  3. 3.Plant Biotechnology Research CenterSchool of Agriculture and Biology, Shanghai Jiao Tong UniversityShanghaiPeople’s Republic of China

Personalised recommendations