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

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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.

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Abbreviations

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

References

  • 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 36–39

    Google Scholar 

  • 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 55–60

    Google Scholar 

  • 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 411–441

    Google Scholar 

  • 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 329–373

    Google Scholar 

  • 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 11–24

    Article  CAS  Google Scholar 

  • 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. 13 165–177

    Article  CAS  Google Scholar 

  • 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 1035–1045

    Article  CAS  Google Scholar 

  • 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 1–11

    Article  CAS  Google Scholar 

  • Escudero J, Neuhaus G, Schlappi M and Hohn B 1996 T-DNA transfer in meristematic cells of maize provided with intracellularAgrobacterium;Plant J. 10 355–360

    Article  CAS  Google Scholar 

  • 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 13–22

    Article  CAS  Google Scholar 

  • 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. 39 967–978

    Article  Google Scholar 

  • 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. 79 295–307

    Article  Google Scholar 

  • Habibi J, Backus E A and Czalpa T H 1993 Plant lectins affect survival of the potato leafhopper (Homoptera: Cicadellidae);J. Econ. Entomol. 86 945–951

    Article  CAS  Google Scholar 

  • 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) 330 160–163

    Article  CAS  Google Scholar 

  • 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 18–25

    Article  CAS  Google Scholar 

  • Hu Q A 2000Agrobacterium-mediated transformation of rice hybrid parental lines using snowdrop lectin gene (gna); Doctoral dissertation, Fudan University, Shanghai, China

    Google Scholar 

  • 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. 14 745–750

    Article  CAS  Google Scholar 

  • Iyer L M, Kumpatla S P, Chandrasekharan M B and Hall T C 2000 Transgene silencing in monocots;Plant Mol. Biol. 43 323–346

    Article  CAS  Google Scholar 

  • 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 20–28

    Article  CAS  Google Scholar 

  • 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. 46 563–565

    Article  CAS  Google Scholar 

  • Matzke M A, Scheid O M and Matzke A J 1999 Rapid structural and epigenetic changes in polyploid and aneuploid genomes;Bioessays 21 761–767

    Article  CAS  Google Scholar 

  • 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. 66 119–126

    Article  CAS  Google Scholar 

  • 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 51–59

    Article  CAS  Google Scholar 

  • 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. 44 529–539

    Article  CAS  Google Scholar 

  • 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. 166 141–149

    Article  CAS  Google Scholar 

  • Rahb Y and Febvay G 1993 Protein toxicity to aphids: anin vitro test onAcyrthosiphon pisum;Entomol. Exp. Appl. 67 149–160

    Article  Google Scholar 

  • 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. 15 469–477

    Article  CAS  Google Scholar 

  • 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. 79 285–293

    Article  CAS  Google Scholar 

  • Schlappi M and Hohn B 1992 Competence of immature maize embryos forAgrobacterium-mediated gene transfer;Plant Cell 4 7–16

    Article  Google Scholar 

  • Sharma H C and Ortiz R 2000 Transgenics, pest management, and the environment;Curr. Sci. 79 421–437

    CAS  Google Scholar 

  • 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 1488–1492

    Article  CAS  Google Scholar 

  • 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. 45 623–631

    Article  CAS  Google Scholar 

  • 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 65–73

    Article  CAS  Google Scholar 

  • 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. 7 371–378

    Article  CAS  Google Scholar 

  • 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 1108–1113

    Google Scholar 

  • 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 208 552–563

    Article  CAS  Google Scholar 

  • 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 14–16

    Google Scholar 

  • 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 2068–2075

    Article  Google Scholar 

  • 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. 47 873–880

    Article  CAS  Google Scholar 

  • 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. 39 155–181

    Google Scholar 

  • 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. 42 719–723

    CAS  Google Scholar 

  • Xiao S H, Liu J G and Huang J Q 2001 Breeding of insecticidal transgenic cotton cultivations;Chin. Cotton 28 19–20

    Google Scholar 

  • Zhang G X and Zhong T S 1983Economic insect fauna of China 1st edition, Vol. 25 (Beijing: Science Press) pp 1–65

    Google Scholar 

  • Zhang X G 1992The research on the resistance of soybean cultivars to Cercospora sojina Hara, Doctoral dissertation, North-East Agricultural University, Harbin, Heilongjiang, China

    Google Scholar 

  • 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. 44 789–798

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Life Science School, Shandong University, 250100, Jinan, People’s Republic of China

    Zhaoyu Wang, Kewei Zhang & Juren Zhang

  2. State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, 200433, Shanghai, People’s Republic of China

    Xiaofen Sun & Kexuan Tang

  3. Plant Biotechnology Research Center, School of Agriculture and Biology, Shanghai Jiao Tong University, 200030, Shanghai, People’s Republic of China

    Kexuan Tang

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  1. Zhaoyu Wang
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  2. Kewei Zhang
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  3. Xiaofen Sun
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  4. Kexuan Tang
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  5. Juren Zhang
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Correspondence to Juren Zhang.

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Wang, Z., Zhang, K., Sun, X. et al. Enhancement of resistance to aphids by introducing the snowdrop lectin genegna into maize plants. J. Biosci. 30, 627–638 (2005). https://doi.org/10.1007/BF02703563

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