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Production and analysis of transgenic wheat (Triticum aestivum L.) with improved insect resistance by the introduction of cowpea trypsin inhibitor gene

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Abstract

The cowpea trypsin inhibitor gene (CpTI) and neomycin phosphotransferase gene (nptII) were introduced into the embryonic callus cells of immature embryos of wheat elite line Shannong 995604 using Agrobacterium-mediated gene transfer. Independent plantlets were regenerated from kanamycin-resistant calli. PCR and real time PCR analysis, PCR-Southern and Southern blot hybridization indicated that there were three independently-dervied transgenic plants viz. transformed-I, II and III (T-I, T-II and T-III). The segregation of CpTI in the transgenic wheat progenies of T-Iand T-III were consistent with Mendelian inheritance. Resistance to the storage insect pest of wheat viz. the grain moth (Sitotroga cerealella Olivier) was improved significantly in seeds of the three transgenic wheat T2 lines obtained from T1 PCR-positive plants. The frequency of moth-eaten seed from T-I, T-IIand T-III was reduced 66.76%, 62.48% and 43.59% respectively. The investigation of agronomic traits of the three transgenic wheat T1 PCR-positive plants revealed that the three transgenic lines had excellent agronomic traits. They provide good germplasm resource for wheat genetic improvement.

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Abbreviations

AS:

acetosyringone

CpTI:

cowpea trypsin inhibitor

CpTI:

cowpea trypsin inhibitor gene

NPTII:

neomycin phosphotransferase gene

CH:

casein hydrolysate

2,4-D-2,4:

dichlorophenoxyacetic acid

ABA:

abscisic acid

KT:

kinetin

Km:

kanamycin

Cef:

Cefotaxime

MS:

medium-Murashige and Skoog medium

Rif:

rifampicin

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Authors and Affiliations

  1. College of Agronomy, Shandong Agricultural University, Subcenter of National Wheat Improvement Center, The Key Laboratory of Crop Biology of Shandong, Taian, 271018, China

    Hai-Yan Jia, De-Shun Feng & Hong-Gang Wang

  2. Department of Life Science, Heze College, Heze, 274015, China

    Rui-Ming Bi

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  1. Rui-Ming Bi
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  2. Hai-Yan Jia
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  3. De-Shun Feng
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  4. Hong-Gang Wang
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Correspondence to Hong-Gang Wang.

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Bi, RM., Jia, HY., Feng, DS. et al. Production and analysis of transgenic wheat (Triticum aestivum L.) with improved insect resistance by the introduction of cowpea trypsin inhibitor gene. Euphytica 151, 351–360 (2006). https://doi.org/10.1007/s10681-006-9157-9

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