Bai, Shiqie

Li, Daxu

Liu, Gang

You, Minghong

English

University of Kentucky

University of Kentucky UKnowledge International Grassland Congress Proceedings XXI International Grassland Congress / VIII International Rangeland Congress Agrobacterium‐Mediated Genetic Transformation of Elymus breviaristatus with Pseudomonas Pseudoalcaligenes Insecticidal Protein Gene Daxu Li Sichuan Academy of Grassland Science, China Shiqie Bai Sichuan Academy of Grassland Science, China Gang Liu Sichuan Academy of Grassland Science, China Minghong You Sichuan Academy of Grassland Science, China Follow this and additional works at: https://uknowledge.uky.edu/igc  Part of the Plant Sciences Commons, and the Soil Science Commons This document is available at https://uknowledge.uky.edu/igc/21/12-1/10 The XXI International Grassland Congress / VIII International Rangeland Congress took place in Hohhot, China from June 29 through July 5, 2008. Proceedings edited by Organizing Committee of 2008 IGC/IRC Conference Published by Guangdong People's Publishing House This Event is brought to you for free and open access by the Plant and Soil Sciences at UKnowledge. It has been accepted for inclusion in International Grassland Congress Proceedings by an authorized administrator of UKnowledge. For more information, please contact UKnowledge@lsv.uky.edu. brought to you by COREView metadata, citation and similar papers at core.ac.ukprovided by University of Kentucky　 Multifunctional Grasslands in a Changing World 　 Volume Ⅱ 　 瞯 ]341　 瞯Grasslands/Rangelands Production Systems——— Developing Improved PlantsAgrobacterium‐mediated genetic transformation of Elymus breviaristatus with pseudomonaspseudoalcaligenes insecticidal protein geneL i Daxu , Bai Shiqie , L iu Gang , You MinghongSichuan Grassland Science A cademy , Chengdu , 611731 , China . E‐mail : ldx136＠ 163 .comKey words :Agrobacterium‐mediated transformation ,callus regeneration ,mature embryo ,ppip ,transgenic plantIntroduction Elymus brev iaristatus is the predominant type of pasture grass in cold season . This forage grass , however , is afavorable food for grasshoppers( Locusta ,Orthop tera) . Grasshopper plague has become a major damage to the maintenance ofpasturage . The development of transgenic technology provides an alternative and effective method for reducing the damage ofgrasshoppers by the products of pesticidal genes ( Akutsu et al , ２００４) .Materials and methods Embryogenic calli of Elymus brev iaristatus were induced from mature seeds , and proliferated on callusinduction and maintenance medium containing kinetin ０ .０５ mg/ L and ２ , ４‐D ８ .０ mg /L under dim‐light condition at ２６ ℃ . Theembryogenic calli produced were used for transformation mediated by EHA１０５ which carries the pCAMBIA １３０４‐ppip . Thecalli transformed were selected and grown in the presence of ６０ mg / L hygromycin ( Li et al , ２００６) .Results MS medium was more favorable for callus induction and regeneration of Elymus brev iaristatus than other media whenmature embryos of seeds were used as explants ( data not shown ) . The calli of three‐and four‐month‐old what had higherfrequency of hygromycin‐resistant callus when co‐cultivated at １９‐２３ ℃ ( data not shown) . The transformation frequency of thecalli reached １０ .５２ ％ ( data not shown) . Calluses induction and regeneration of Elymus brev iaristatus are shown in Figure １ .The presence of p p ip gene in the genomic DNA of regenerated plants was detected by means of Southern hybridization ( Figure２ ) , and the expression of the trans‐genes was verified by reverse transcription‐PCR ( Figure ３ ) .Figure1 A lluses induction andregenerationo f Elymus breviaristatus .　Figure 2 DNA gel blots analysis o ftransgenic Elymus brev iaristatus .　Figure 3 RT‐PCR analysis o f transgenicElymus brev iaristatus .Conclusions A grobacterium‐mediated transformation in theconditions tested was effective for transferring foreign genes intoElymus brev iaristatus . These results may set an example for the A grobacterium method that can effectively transfer foreigngenes into Elymus brev iaristatus .ReferencesAkutsu M , Ishizaki T , Sato H (２００４) T ransformation of the monocotyledonous Alstroemeria by Agrobacterium tumefaciens .Plant Cell Rep . ２２(８) : ５６１‐８ .Li DX , Zhang J , Zhao J , Zhang Y , L１ L , Liu SJ , Chen F , Yang ZR (２００６) . T ransformation of calli of siberian wildrye grass( elymus sibiricus L . cv . chuancao No .２ ) mediated by agrobacterium . Journal o f Plant Physiology and MolecularBiology . ２００６ , ３２ (１) : ４５‐５１ .

Agrobacterium‐Mediated Genetic Transformation of \u3ci\u3eElymus breviaristatus\u3c/i\u3e with Pseudomonas Pseudoalcaligenes Insecticidal Protein Gene

https://uknowledge.uky.edu/cgi/viewcontent.cgi?article=3265&amp;context=igc

Agrobacterium‐Mediated Genetic Transformation of \u3ci\u3eElymus breviaristatus\u3c/i\u3e with Pseudomonas Pseudoalcaligenes Insecticidal Protein Gene

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