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Pyramiding of Bt cry1Ia8 and cry1Ba3 genes into cabbage (Brassica oleracea L. var. capitata) confers effective control against diamondback moth

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Abstract

A binary vector carrying Bacillus thuringiensis cry1Ia8 and cry1Ba3 genes was introduced into an inbred line of white cabbage by Agrobacterium tumefaciens-mediated transformation for control of diamondback moth (DBM, Plutella xylostella), and 14 kanamycin-resistant plantlets were obtained. Presence and expression of the cry1Ia8 and cry1Ba3 genes in five transformed plants were confirmed by polymerase chain reaction (PCR), Southern blot, reverse transcription-polymerase chain reaction (RT-PCR), and Western blot analyses. Insect bioassays showed that these transgenic plants were able to effectively control both susceptible and Cry1Ac-resistant DBM larvae as compared to non-transformed counterparts. Ten homozygous insect-resistant cabbage lines were obtained in the T2 generation through self-pollination, and molecular methods and insect bioassays. After natural infestation under greenhouse and field conditions, the pyramided lines exhibited excellent efficacy against DBM. Furthermore, data from field trials indicated that there were no significant differences in most agronomic traits between most the homozygous lines and the original variety. These transgenic lines may allow field study of resistance management strategies involving gene pyramiding and serve as novel insect-resistant resources in cabbage breeding.

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Acknowledgments

We are grateful to Prof. Dafang Huang, Prof. Jie Zhang, and Dr. Zhihong Lang for providing Bt genes and vector. We thank Prof. Youjun Zhang for their help in sending us the DBM larvae. Thanks are due to Prof. Elizabeth Earle (Department of Plant Breeding & Genetics, Cornell University, USA) for constructive advice on writing the manuscript and to Dr. Jun Cao (Athenix Corporation, USA) for providing the Brassica genetic transformation protocol. This work was supported by grants from the National High Technology Research and Development Program of China (863 Program, No. 2008AA10Z155), the National Natural Science Foundation of China (No. 31071697), and the earmarked fund for Modern Agro-industry Technology Research System (No. nycytx-35-gw01).

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Authors and Affiliations

  1. College of Agronomy and Biotechnology, China Agricultural University, Beijing, 100193, People’s Republic of China

    Dengxia Yi & Zhenxian Zhang

  2. Key Laboratory of Biology and Genetic Improvement of Horticultural Crops, Ministry of Agriculture, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, 100081, People’s Republic of China

    Dengxia Yi, Lei Cui, Li Wang, Yumei Liu, Mu Zhuang, Yangyong Zhang, Zhiyuan Fang & Limei Yang

  3. Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, 100193, People’s Republic of China

    Jie Zhang

  4. Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing, 100081, People’s Republic of China

    Zhihong Lang

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  1. Dengxia Yi
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Correspondence to Limei Yang.

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Yi, D., Cui, L., Wang, L. et al. Pyramiding of Bt cry1Ia8 and cry1Ba3 genes into cabbage (Brassica oleracea L. var. capitata) confers effective control against diamondback moth. Plant Cell Tiss Organ Cult 115, 419–428 (2013). https://doi.org/10.1007/s11240-013-0373-4

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  • DOI: https://doi.org/10.1007/s11240-013-0373-4

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