Agrobacterium-mediated transformation of spinach (Spinacia oleracea) with Bacillus thuringiensis cry1Ac gene for resistance against two common vegetable pests

スポンサーリンク

概要

• 論文の詳細を見る

• Agrobacterium-mediated genetic transformation was applied to produce transgenic plants of spinach (Spinacia oleracea) resistant to 2 pest species, Trichoplusia ni and Autographa nigrisigna. Leaf segments from in vitro spinach plants of cultivar ‘Glory’ were co-cultivated with A. tumefaciens strain EHA105, which harbored the plasmid pBE2111FMB containing a synthetic cry1Ac gene encoding an insecticidal crystal protein of Bacillus thuringiensis, and neomycin phosphotransferase II and bialaphos resistance genes as selectable marker genes. Kanamycin (Km)-resistant calluses were obtained from co-cultivated leaf segments 1 month after selection on 2.5 g l−1 gellan gum-solidified MS medium containing 30 g l−1 sucrose, 0.5 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D), 2 mg l−1 kinetin, 100 mg l−1 Km and 20 mg l−1 meropenem trihydrate. Regeneration of adventitious shoots from Km-resistant calluses was performed on 2.5 g l−1 gellan gum-solidified MS medium containing 30 g l−1 sucrose, 0.01 mg l−1 2,4-D, 1 mg l−1 kinetin, 1 mg l−1 gibberellic acid, 100 mg l−1 Km and 10 mg l−1 meropenem trihydrate at 14°C. PCR and Southern blot analyses of genomic DNA isolated from T1 plants confirmed the successful integration of T-DNA into the plant genome. Expression of Cry1Ac protein was confirmed in leaves of transgenic plants by Western blot analysis. Insect bioassays against T. ni and A. nigrisigna performed with T1 plants showed more than 93.3% insect mortality within 1 week. These results suggest that the cry1Ac gene was effectively expressed in spinach.

• 日本植物細胞分子生物学会の論文
• 2009-03-25

著者

• Chin Dong

  Graduate School Of Horticulture Chiba Univ.

• Chin Dong

  Lab. Of Plant Cell Technol. Graduate School Of Horticulture Chiba Univ.

• MII Masahiro

  Graduate School of Horticulture, Chiba University

• Bao Jin

  Laboratory Of Plant Cell Technology Graduate School Of Horticulture Chiba University

• BAO Jin

  Graduate School of Horticulture, Chiba University

• FUKAMI Masanobu

  Chiba Prefectural Agriculture and Forestry Research Center

• UGAKI Masashi

  Graduate School of Frontier Sciences, The University of Tokyo

• NOMURA Masashi

  Graduate School of Horticulture, Chiba University

• Mii Masahiro

  Graduate School Of Horticulture Chiba University

• Ugaki Masashi

  Graduate School Of Frontier Sciences The University Of Tokyo

• Nomura Masashi

  Graduate School Of Horticulture Chiba University

• Mii Masahiro

  Graduate School Of Horticulture Chiba Univ.

関連論文

• Transgenic spinach plants produced by Agrobacterium-mediated method based on the low temperature-dependent high plant regeneration ability of leaf explants
• Generation of phenotypically normal marker-free transgenic plants of Kalanchoe blossfeldiana through hairy root induction
• Thidiazuron-induced rapid shoot regeneration via embryo-like structure formation from shoot tip-derived callus culture of sugarcane
• Agrobacterium-mediated transformation of spinach (Spinacia oleracea) with Bacillus thuringiensis cry1Ac gene for resistance against two common vegetable pests
• Gametosomatic hybridization between egg cell protoplast and mesophyll protoplast of Petunia hybrida
• Direct isolation of female germ units from ovules of Petunia hybrida by enzymatic treatment without releasing somatic protoplasts from ovular tissue
• Transgenic Phalaenopsis plants with resistance to Erwinia carotovora produced by introducing wasabi defensin gene using Agrobacterium method
• Rapid and simple DNA extraction method from rice using a glass-fiber filter inserted pipette tip
• Transgenic potatoes expressing wasabi defensin peptide confer partial resistance to gray mold (Botrytis cinerea)
• Effect of heavy ion-beam irradiation on plant growth and mutation induction in Nicotiana tabacum
• Unreduced gametes are the major causal factor for the production of polyploid interspecific hybrids in Primula
• Histological observation of root bud formation of hairy roots in Lotus corniculatus L.
• Recent advances in wide hybridization and embryo rescue of floricultural plants in Japan
• Thidiazuron-induced rapid shoot regeneration via embryo-like structure formation from shoot tip-derived callus culture of sugarcane
• Agrobacterium-mediated genetic transformation of Cattleya with an Odontoglossum ringspot virus replicase gene sequence
• Overproduction of artemisinin in tetraploid Artemisia annua L.
• Phylogenetic classification of the phytoplasma: Identification of a new phytoplasma from diseased chestnut tree in Korea (第28回日本マイコプラズマ学会学術集会)
• Cloning of oligopeptide permiase homologues from OY phytoplasma genome (第28回日本マイコプラズマ学会学術集会)
• Interspecific hybridization of Begonia semperflorens (section Begonia) with B. pearcei (section Eupetalum) for introducing yellow flower color
• Ionic compositions play an important role on in vitro propagation of PLBs of spring-flowering Calanthe
• Plant regeneration from Crotalaria spectabilis hairy roots which showed inhibited growth of root-knot nematodes (Meloidogyne hapla and M. incognita) in vitro
• Transgenic Petunia hybrids expressing a synthetic fungal chitinase gene confers disease tolerance to Botrytis cinerea
• Highly efficient system for plant regeneration from leaf and stem explants in Dahlia
• Establishment of Agrobacterium-mediated genetic transformation system in Dahlia
• Characterization of lignocellulose of Erianthus arundinaceus in relation to enzymatic saccharification efficiency
• Efficient plant regeneration system from seed-derived callus of ravenna grass [Erianthus ravennae (L.) Beauv.]
• Characterization of lignocellulose of Erianthus arundinaceus in relation to enzymatic saccharification efficiency

もっと見る 閉じる

スポンサーリンク