Lu, Xiaoping

Mi, Fugui

Wang, Shuyan

Xiao, Yuhong

Yin, Li

English

University of Kentucky

University of Kentucky UKnowledge International Grassland Congress Proceedings XXI International Grassland Congress / VIII International Rangeland Congress Genetic Transformation of Tomato Pollen Lysine‐Rich Gene to Elytrigia intermedium and Regeneration of Transgenic Plants Xiaoping Lu Inner Mongolia Agricultural University, China Li Yin Inner Mongolia Agricultural University, China Fugui Mi Inner Mongolia Agricultural University, China Yuhong Xiao Chinese Academy of Sciences, China Shuyan Wang Inner Mongolia Agricultural University, 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/21/12-2/1 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. 瞯 ]382　 瞯 　 Multifunctional Grasslands in a Changing World 　 Volume Ⅱ 　Grasslands/Rangelands Production Systems——— Developing Improved PlantsGenetic transformation of tomato pollen lysine‐rich gene to Elytrigia intermedium andregeneration of transgenic plantsX iaop ing Lu1 , L i Y in1 , Fugui M i1 , Y uhong X iao2 , Shuyan W ang11 College o f A gron . , Inner Mongolia A gric . Univ . , H uhhot , Inner Mongolia 010018 P .R . o f China ; 2 Bei j ing Gen .A cad . in Sci . A cad . o f China , Bei j ing 101300 . 1 E‐mail :lx p1960＠ 163 .comKey words : Ely trigia intermedia ,lysine‐rich gene ,transformation ,regenerationIntroduction Elytrigia intermedium ( Hest ) Nevski is a perennial T riticeae grass that has high biomass production , highpalatability and digestion , but low lysine content . A pollen‐specific lysine‐rich gene cDNA sequence from tomato was obtainedby RT‐PCR . A plant expression plasmid pBIUB‐TSB was constructed with the ubiquitin promoter . Plant regeneration wasconstructed using the mature seed of Elytrigia intermedia as the explant . Plasmids of pBIUB‐TSB were transferred into thecalli by the particle gun and the laser microbeam methods . The regenerated plants were selected for kanamycin resistance .Materials and methods The plant regeneration of E . intermedium was accomplished by use of a : １) calli inducing medium of MS＋ ２ .４‐D (４ .５mg / L) ＋ BA ０ .１mg /L ＋ Sucrose ３０g / L ＋ Agar ６ .５g / L PH :５ .８ ～ ６ .０ , ２) subculture medium of １) ＋ Vc ５mg/ L ,３ ) differentiation medium of MS ＋ BA ２mg /L ＋ NAA １mg / L ＋ CoCl２ ５mg /L ＋ ZT ０ .５mg/ L , and ４ ) rooting medium of １ /２MS ＋ IAA ０ .５mg /L ＋ NAA ０ .５mg/ L ＋ AC ０ .１％ . The particle gun and the laser microbeam methods were used intransformation .Results Some regenerated plants were obtained by both the particle gun and laser microbeam under selection for kanamycinresistance of ３００mg /L . A total of １４ plants were expressed positively by genomic PCR detection and southern blottinganalysis , ９ plants were transformed with the particle gun , and ５ plants were transformed with the laser microbeam ( Figure １ ,Figure ２) .Figure 1 Electropherogram o f PCR o f transgenic p lants : 1 ＝ DNA marker , 2 ＝ positive control ; 3 ＝ negative control , 4 － 7 ＝particle gun trans f ormed p lants , and 8 ,9 ＝ laser microbeam trans f ormed p lants .Figure 2 Southern blot o f transgene p lants :1 ＝ p lasmid pBIUB － TSB , 2 ＝ negative control ,3 ,4 ＝ trans f ormed p lants w iththe particle gun method , 5 ,6 ＝ trans f ormed p lants w ith the laser microbeam method .Conclusions A regeneration system of E . intermedium ( Hest) Nevski was obtained with seed explant . Both the particle gun andthe laser microbeam methods were used successfully in transformation of calli of E . intermedium . ; PCR and southern blottingproved that the lysine‐rich gene was integrated into the genome of E . intermedium .ReferencesShirsat , A . , Wild , N .R .C . , Bouter , D . , １９８９ . Sequences responsible for the tissue specific promoter activity of a pealegumin gene in tobacco . Mol . Gen . Genet . ２１５ , ３２６‐３３１ .Zhao , Q . , Yu , J .J . , Zhu , D .Y . , Ao , G .M . , ２００４ . Cloning and characterization of a pollen‐specific lysine‐rich proteincDNA ( tsb) from tomato . Chin . J . Biochem . &Mol . Biol . ２０ (２) , ２７５‐２７９ .

Genetic Transformation of Tomato Pollen Lysine‐Rich Gene to \u3ci\u3eElytrigia intermedium\u3c/i\u3e and Regeneration of Transgenic Plants

https://uknowledge.uky.edu/cgi/viewcontent.cgi?article=3306&amp;context=igc

Genetic Transformation of Tomato Pollen Lysine‐Rich Gene to \u3ci\u3eElytrigia intermedium\u3c/i\u3e and Regeneration of Transgenic Plants

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