caa a22 4500 463236378 CHVBK 20180405153258.0 cr unu---uuuuu 170326e20070501xx s 000 0 eng 10.1007/s11240-007-9229-0 doi (NATIONALLICENCE)springer-10.1007/s11240-007-9229-0 Agrobacterium -mediated genetic transformation of Elymus breviaristatus with Pseudomonas pseudoalcaligenes insecticidal protein gene [Elektronische Daten] [Daxu Li, Qun Sun, Min Huang, Jie Zhang, Shiqie Bai, Linyong Zheng, Jian Zhao, Dunlian Qiu, Li Li, Zhirong Yang, Minghong You, Gang Liu, Yu Zhang, Changbing Zhang, Shaocai Li] An efficient and repeatable protocol for generating transgenic Elymus breviaristatus plants was first established by Agrobacterium-mediated transformation: calli of Elymus breviaristatus were induced from mature seeds, and proliferated on callus induction and maintenance medium containing kinetin 0.05mg/l and 2, 4-D 8.0mg/l under dim-light condition (10-20μmol/m2s,16h light) at 26°C. The calli produced were used for transformation mediated by Agrobacterium tumefaciens, EHA105, which carries the binary vector pCAMBIA 1304-ppip, coding for the Pseudomonas pseudoalcaligenes insecticidal protein gene (ppip) and hygromycin phosphotransferase II gene (hptII). The calli transformed were selected and grown in the presence of 60mg/l hygromycin. Approximately 90 transgenic plants were produced, and transformation frequency of the calli reached 10.52%. The presence of ppip gene in the genomic DNA of regenerated plants was detected by means of PCR and Southern-blot analysis, and the expression of the transgenes was verified by reverse transcription-PCR. Approximately 90% of the tested plants appeared to have only one or two copies of the T-DNA inserts. Verification of anti-grasshoppers activity shows the mortality of grasshoppers reached 16.73%. These results indicated that Agrobacterium-mediated transformation in our condition was effective for transferring foreign genes into Elymus breviaristatus. Springer Science+Business Media B.V., 2007 Agrobacterium -mediated transformation nationallicence Callus regeneration nationallicence Mature embryo nationallicence ppip gene nationallicence Transgenic plant nationallicence ABA : abscisic acid nationallicence AS : acetosyringone nationallicence 6-BA : 6-benzylaminopurine nationallicence 2,4-D : 2,4-dichlorophenoxyacetic acid nationallicence hpt II : hygromycin phosphotransferase II nationallicence MS : Murashige and Skoog medium nationallicence NAA : 1-naphthaleneacetic acid nationallicence ppip : seudoalcaligenes insecticidal protein gene nationallicence Li Daxu Sichuan Academy of Grassland Science, 611731, Chengdu, P.R. China aut Sun Qun College of Life Sciences, Key Laboratory of Bio-resources and Eco-environment, The Ministry of Education, Sichuan University, 610064, Chengdu, P.R. China aut Huang Min College of Life Sciences, Key Laboratory of Bio-resources and Eco-environment, The Ministry of Education, Sichuan University, 610064, Chengdu, P.R. China aut Zhang Jie College of Life Sciences, Key Laboratory of Bio-resources and Eco-environment, The Ministry of Education, Sichuan University, 610064, Chengdu, P.R. China aut Bai Shiqie Sichuan Academy of Grassland Science, 611731, Chengdu, P.R. China aut Zheng Linyong College of Life Sciences, Key Laboratory of Bio-resources and Eco-environment, The Ministry of Education, Sichuan University, 610064, Chengdu, P.R. China aut Zhao Jian College of Life Sciences, Key Laboratory of Bio-resources and Eco-environment, The Ministry of Education, Sichuan University, 610064, Chengdu, P.R. China aut Qiu Dunlian College of Life Sciences, Key Laboratory of Bio-resources and Eco-environment, The Ministry of Education, Sichuan University, 610064, Chengdu, P.R. China aut Li Li College of Life Sciences, Key Laboratory of Bio-resources and Eco-environment, The Ministry of Education, Sichuan University, 610064, Chengdu, P.R. China aut Yang Zhirong College of Life Sciences, Key Laboratory of Bio-resources and Eco-environment, The Ministry of Education, Sichuan University, 610064, Chengdu, P.R. China aut You Minghong Sichuan Academy of Grassland Science, 611731, Chengdu, P.R. China aut Liu Gang Sichuan Academy of Grassland Science, 611731, Chengdu, P.R. China aut Zhang Yu Sichuan Academy of Grassland Science, 611731, Chengdu, P.R. China aut Zhang Changbing Sichuan Academy of Grassland Science, 611731, Chengdu, P.R. China aut Li Shaocai College of Life Sciences, Key Laboratory of Bio-resources and Eco-environment, The Ministry of Education, Sichuan University, 610064, Chengdu, P.R. China aut Plant Cell, Tissue and Organ Culture Kluwer Academic Publishers 89/2-3(2007-05-01), 159-168 0167-6857 89:2-3<159 2007 89 11240 https://doi.org/10.1007/s11240-007-9229-0 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11240-007-9229-0 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Li Daxu Sichuan Academy of Grassland Science, 611731, Chengdu, P.R. China aut NATIONALLICENCE 700 1- Sun Qun College of Life Sciences, Key Laboratory of Bio-resources and Eco-environment, The Ministry of Education, Sichuan University, 610064, Chengdu, P.R. China aut NATIONALLICENCE 700 1- Huang Min College of Life Sciences, Key Laboratory of Bio-resources and Eco-environment, The Ministry of Education, Sichuan University, 610064, Chengdu, P.R. China aut NATIONALLICENCE 700 1- Zhang Jie College of Life Sciences, Key Laboratory of Bio-resources and Eco-environment, The Ministry of Education, Sichuan University, 610064, Chengdu, P.R. China aut NATIONALLICENCE 700 1- Bai Shiqie Sichuan Academy of Grassland Science, 611731, Chengdu, P.R. China aut NATIONALLICENCE 700 1- Zheng Linyong College of Life Sciences, Key Laboratory of Bio-resources and Eco-environment, The Ministry of Education, Sichuan University, 610064, Chengdu, P.R. China aut NATIONALLICENCE 700 1- Zhao Jian College of Life Sciences, Key Laboratory of Bio-resources and Eco-environment, The Ministry of Education, Sichuan University, 610064, Chengdu, P.R. China aut NATIONALLICENCE 700 1- Qiu Dunlian College of Life Sciences, Key Laboratory of Bio-resources and Eco-environment, The Ministry of Education, Sichuan University, 610064, Chengdu, P.R. China aut NATIONALLICENCE 700 1- Li Li College of Life Sciences, Key Laboratory of Bio-resources and Eco-environment, The Ministry of Education, Sichuan University, 610064, Chengdu, P.R. China aut NATIONALLICENCE 700 1- Yang Zhirong College of Life Sciences, Key Laboratory of Bio-resources and Eco-environment, The Ministry of Education, Sichuan University, 610064, Chengdu, P.R. China aut NATIONALLICENCE 700 1- You Minghong Sichuan Academy of Grassland Science, 611731, Chengdu, P.R. China aut NATIONALLICENCE 700 1- Liu Gang Sichuan Academy of Grassland Science, 611731, Chengdu, P.R. China aut NATIONALLICENCE 700 1- Zhang Yu Sichuan Academy of Grassland Science, 611731, Chengdu, P.R. China aut NATIONALLICENCE 700 1- Zhang Changbing Sichuan Academy of Grassland Science, 611731, Chengdu, P.R. China aut NATIONALLICENCE 700 1- Li Shaocai College of Life Sciences, Key Laboratory of Bio-resources and Eco-environment, The Ministry of Education, Sichuan University, 610064, Chengdu, P.R. China aut NATIONALLICENCE 773 0- Plant Cell, Tissue and Organ Culture Kluwer Academic Publishers 89/2-3(2007-05-01), 159-168 0167-6857 89:2-3<159 2007 89 11240 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer