|
|
|
| The Genetic Transformation of Peanut (Arachis hypogaea L.) Cotyledon Explants Using Xylose as Screening Reagent |
|
|
|
|
Abstract Those antibiotics and herbicides marker genes generally used at present in plant genetic transformation might be potential treats and dangers for human being and environment, so it is urgent to search and develop new secure screening reagents instead of antibiotics and herbicides reagents. New recombinant plasmid pCAMBIA1301-xylA-Ah-Glu-P with promoter of peanut (Arachis hypogaea L.) β-1,3-glucanase gene (Ah-Glu-P) and xylose isomerase gene (xylA) from Escherichia coli was constructed successfully. pCAMBIA1301-xylA-Ah-Glu-P was then transformed to cotyledon explants of peanut variety Huayu23 by Agrobacterium mediated method using xylose as screening reagent. The frequency of regeneration seedling was 16.1% and the frequency of transformed positive plants reached to 78.4%, when the concentration of sucrose and xylose in medium was 10 and 20 g/L, respectively. The transgenic positive plants were then transplanted to field by grafting and domesticating and the survival rate of transplanted plants was up to 74.5%. The transformation method with cotyledons as explants, with xylose as screening reagent, has characteristics of simple and rapid, safe and pollution-free and higher transgenic positive frequency. So it was a safe and efficient genetic transformation method compared with other transformation methods using other explants and antibiotics screening reagent.
|
|
Received: 12 January 2015
Published: 09 July 2015
|
|
Corresponding Authors:
lixian qiao
E-mail: lxqiao73@163.com
|
|
|
|
| [1]Miki B, McHugh S.Selectable marker genes in transgenic plants: applications,alternatives and biosafety[J].J Biotechnol, 2004, 107(3):193-232
[2] 刘广金, 姚晓林, 蒋家平等.转基因植物中安全标记基因的研究进展[J].现代农业科技, 2008, (4):208-208
[3]Jaiwal PK, Sahoo L, Singh ND.Strategies to deal with the concern about marker genes in transgenic plants:some environment-friendly approaches[J].Current science, 2002, 83(2):128-136
[4] 刘风珍, 万勇善, 潘昱名.花生遗传转化高效基因型的筛选与研究[J].花生学报, 2009, 38(4):21-25
[5] 李拴柱.花生维生素E合成关键酶基因的克隆、序列分析及γ-TMT基因的转化研究[D]. 山东农业大学, 2012
[6] 高明, 谭化, 王云鹏等.根癌农杆菌介导花生遗传转化体系的建立[J].吉林农业科学, 2013, 38(3):35-38
[7] 贾宇臣, 王利, 陈琦等.花生子叶节外植体遗传转化体系的建立[J].科学技术与工程, 2011, 11(14):3142-3146
[8] 王允.花生子叶再生体系及转化体系的建立与优化[D]. 河南农业大学, 2012.
[9]Lucca P, Ye X, Potrykus I.Effective selection and regeneration of transgenic rice plants with mannose as selective agent[J].Mol Breed, 2001, 7(1):43-49
[10] Wang AS, Evans RA, Altendorf PR.A mannose selection system for production of fertile transgenic maize plants from protoplasts[J].Plant Cell Reports, 2000, (19):654-660
[11]Wright M, Dawson J, Dunder E et al..Efficient biolistic transformation of maize ( Zea mays L.) and wheat ( Triticum aestivum L.) using the phosphomannose isomerase gene, pmi , as the selectable marker[J].Plant Cell Reports, 2001, (20):429-436
[12]Joersbo M, Petersen SG, Okkels FT.Parameters interacting with mannose selection employed for the production of transgenic sugar beet[J].Physiologia Plantarum, 1999, 105(1):109-115
[13]岳建雄, 孟钊红, 张炼辉等.以甘露糖作为筛选剂的棉花遗传转化[J].棉花学报, 2005, 17(1):3-7
[14] 彭世清, 陈守才.甘露糖阳性选择系统的建立及在番茄转化中的应用[J]. ,, (2): 141-144[J].农业生物技术学报, 2005, 13(2):141-144
[15] 杨彩云, 尹伟伦, 夏新莉.甘露糖正向筛选体系的建立及在拟南芥遗传转化中的应用[J].分子植物育种, 2009, 7(6):1120-1129
[16] 韦正乙.利用木糖选择系统获得HAL1转基因百脉根（Lotus cirniculatus）植株及耐盐性鉴定[D]. 东北师范大学, 2007
[17]Haldrup A, Petersen SG, Okkels FT.The xylose isomerase gene from Thermoanaerobacterium thermosulfurogenes allows effective selection of transgenic plant cells using D-xylose as the selection agent[J].Plant Molecular Biology, 1998, 37(2):287-296
[18] 崔丽巍.木糖筛选系统在黄瓜转化中的应用[D]. 东北师范大学, 2010
[19]郭新梅, 张晓东, 梁荣奇，等.以木糖异构酶基因为筛选标记的玉米遗传转化[J].植物生理与分子生物学学报, 2007, 33(6):547-552
[20]丁霄, 隋炯明, 王晶珊, 等.以木糖异构酶基因作为选择标记的花生遗传转化[J].核农学报, 2012, 26(3):0444-0448
[21]郝世俊, 隋炯明, 乔利仙, 王晓杰, 王晶珊.花生组培苗嫁接技术的研究[J].青岛农业大学学报自然科学版, 2010, 27(2):110-113
[22] 王秀峰, 王学国, 马艺荞等.木糖筛选剂在辣椒遗传转化上的应用[J].辣椒杂志, 2014, (1):30-32
|
|
|
|
