Dong, Shujun

Huo, Xiuwen

Mi, Fugui

Wang, Guihua

English

University of Kentucky

University of Kentucky UKnowledge International Grassland Congress Proceedings 21st International Grassland Congress / 8th International Rangeland Congress Studies on the Expression of Exogenous p5CS Gene in Transgenic Wheatgrasses Fugui Mi Inner Mongolia Agricultural University, China Guihua Wang Inner Mongolia Agricultural University, China Xiuwen Huo Inner Mongolia Agricultural University, China Shujun Dong 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/21/1-5/34 The 21st International Grassland Congress / 8th 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. 　 Multifunctional Grasslands in a Changing World 　 Volume Ⅰ 　 瞯 ]233　 瞯Grasslands/Rangelands Resources and Ecology ——— Ecology of Grasslands/RangelandsStudies on the expression of exogenous p5CS gene in transgenic wheatgrassesMi f ugui , W ang guihua , H uo X iuwen , Dongshuj unCollege o f Ecology and Env ironment , Inner Mongolia A gricultural University ; H uhhot , ０１００１８ ,ChinaE‐mail : m f gui＠ yahoo .com .cnKey words :Wheatgrasses , T ransgenic , p５CS gene , Northern blotIntroduction T ransgenic wheatgrasses generated from hybrid wheatgrasses ( A gropy ron cristatum × A . desertorum cv .�Hycrest‐Mengnong�) were indentified by PCR analysis and Southern blot . The p５CS gene , which regulates the last step ofproline synthesis in plants , was transferred into wheatgrasses . Northern blot was investigated . Results of Northern blot assaydisplayed that exogenous gene p５CS was expressed at transcription level in transgenic plants . New salt resistance plant lineswhich are adapted to extensive arid and semiarid areas of west China are expected to breed through these processes .Materials and methods Plants of �Hycrest‐Mengnong�wheatgrasses with p５CS gene that have been tested by PCR and southernblot were used as materials and compared to non transgenic plants . Plant total RNA was extracted by kit ; af ter plasmid DNAwas amplified by PCR , arm fragments were extracted by kit as templates . They were labeled with DIG High prime DNALabeling and Detection Starter Kit Ⅰ by the random primer method ; electrophoresed RNA in formol denatured gel ; transferredusing capillary blotting ; hybridized and detected by probe labeled with DIG .Results Figure １ shows the results of Northern blotting of p５CS transgenic plants and negative plants . The hybridization bandof p５CS transgenic plants tested by PCR and Southern blot hybridized with the DIG probe was obvious . It proved thatexogenous gene p５CS is expressed at transcription level in transgenic plants .Figure 1 5 Northern blotting o f transgenic wheatgrass p lant .1 ～ 5 － transgenic p lants ,6 － negative controlConclusions The p５CS catalyse the proline biosynthesis . Its activity , inhibited by proline content , p５CSF１２９A is a mutant wipeoff feedback inhibition of p５CS and it led to a multiple increase of proline content , thus can enhance the protection of plantunder osmotic stress . Now it has been transformed into many plant species as tobacco , rice , ryegrass and tall fescue to enhancethe resistance to drought and salt . The results indicated that the exogenous gene p５CS was expressed at transcription level intransgenic plants .ReferencesHuo Xiuwen , J .Wei , H .Xu , F .Mi , J .Yun . Plant Regeneration and Genetic T ransformation in Wheatgrass . A griculturalsciences in China , ２００６ ,５ (９) : ６４８‐６５４ .Hong Z , Lakkineni K , Zhang Z , Verma D P S . Removal of feedback inhibition of １ pyrroline‐５‐carboxylatesynthetase ( p５CS)results in increased proline accumulation and protection of plant from osmotic stress [ J] . Plant Physiol ,２０００ ,１２２ : １１２９‐１１３６ .2 3 4 5 6 

Studies on the Expression of Exogenous p5CS Gene in Transgenic Wheatgrasses

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Studies on the Expression of Exogenous p5CS Gene in Transgenic Wheatgrasses

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