紫花苜蓿柠檬酸合成酶基因的克隆及对烟草的转化

doi:10.11733/j.issn.1007-0435.2010.04.013

›› 2010, Vol. 18 ›› Issue (4): 550-555.DOI: 10.11733/j.issn.1007-0435.2010.04.013

紫花苜蓿柠檬酸合成酶基因的克隆及对烟草的转化

靳苗苗¹, 杨青川², 金洪¹, 康俊梅², 龙瑞才³

1. 内蒙古农业大学, 呼和浩特, 010019;
2. 中国农业科学院北京畜牧兽医研究所, 北京, 100193;
3. 重庆大学, 重庆, 400044

收稿日期:2010-03-30 修回日期:2010-05-18 出版日期:2010-08-15 发布日期:2010-08-15
通讯作者: 杨青川,qchyang66@yahoo.com.cn,jinhong915@163.com
作者简介:靳苗苗(1984- ),女,内蒙古鄂尔多斯人,硕士研究生,研究方向为植物资源利用与保护方面的研究,E-mail:nancy0194@qq.com.
基金资助:
国家科技支撑计划(2006BAD01A19-3)资助

Cloning of MsCS Gene from Medicago sativa L. and Transformation of Tobaccos

JIN Miao-miao¹, YANG Qing-chuan², JIN Hong¹, KANG Jun-mei², LONG Rui-cai³

1. Inner Mongolia Agricultural University, Hohhot, Inner Mongolia Autonomous Region 010019, China;
2. Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100094, China;
3. Chong Qing University, Chongqing 400044, China

Received:2010-03-30 Revised:2010-05-18 Online:2010-08-15 Published:2010-08-15

摘要/Abstract

摘要： 柠檬酸合成酶是调节植物体内有机酸的活性酶之一,增加植物体内的柠檬酸可以提高植物对土壤中磷的吸收以及抗铝毒性。通过从紫花苜蓿(Medicago sativa L.)中克隆柠檬酸合成酶基因(MsCS),以期获得该基因全长,并为将来研究紫花苜蓿耐铝性以及对磷的吸收方向提供基础。首先利用cDNA末端快速扩增方法(RACE),克隆获得MsCS的cDNA全序列,然后利用pBI121构建植物超表达载体pBI-MsCS,通过农杆菌介导的叶盘法转化烟草(Nicotiana tabacum L.)。结果显示:该基因序列全长为2031 bp,开放阅读框1551 bp,编码517个氨基酸,与其他物种的柠檬酸合成酶具有高度的同源性。获得17株卡那抗性植株。经PCR检测有6株为阳性植株,初步表明该基因已整合到烟草的基因组中。

关键词: 紫花苜蓿, 柠檬酸合成酶基因, 基因克隆, 载体构建, 转化

Abstract: The citrate synthase,an active enzyme,can regulate organic acids in plants.Increasing citric acid can improve plant capabilities of absorbing soil phosphorus and also affect resistance to aluminum toxicity.The gene named MsCS was cloned from alfalfa(Medicago sativa L.) for alfalfa to proceed with basic research of aluminum tolerance and soil absorption for phosphorus.First a full-length cDNA of MsCS was cloned by rapid amplification of cDNA ends(RACE).Then the plant expression vector named pBI-MsCS was constructed on pBI121 vector,and transformed the tobacco by using agrobacterium-mediated.Results indicate that the length of cDNA sequence was 2031 bp and included a 1551 bp open reading frame which encoded a deduced protein of 517-amino-acid polypeptide.The amino acid sequence comparison revealed high homology with citrate synthase of other plants.We obtained a total of 17 kanamycin resistant transgenic plants.PCR detection confirmed 6 positive plants,preliminary studies showed that the MsCS had been integrated into the tobacco genome.

Key words: Medicago sativa L., MsCS gene, Cloning, Vector construction, Transformation

中图分类号:

Q946.5

靳苗苗, 杨青川, 金洪, 康俊梅, 龙瑞才. 紫花苜蓿柠檬酸合成酶基因的克隆及对烟草的转化[J]. , 2010, 18(4): 550-555.

JIN Miao-miao, YANG Qing-chuan, JIN Hong, KANG Jun-mei, LONG Rui-cai. Cloning of MsCS Gene from Medicago sativa L. and Transformation of Tobaccos[J]. , 2010, 18(4): 550-555.

0
/ / 推荐

导出引用管理器 EndNote|Ris|BibTeX

链接本文: https://manu40.magtech.com.cn/Jweb_cdxb/CN/10.11733/j.issn.1007-0435.2010.04.013

https://manu40.magtech.com.cn/Jweb_cdxb/CN/Y2010/V18/I4/550

参考文献

[1] Hanninng I.,Heldt H.W.On the function of mitochondrial metabolism during photosynthesis in spinach leaves (Spinacia oleracea L.)Partitioning between respiration and export of redox equivalents and precursors for nitrate assimilation products[J].Plant Physiology,1993,103:1147-11549
[2] Lo pez-Bucio J,Octavio de la Vega M,Guevara-Garci a A,et al.Enhanced phosphorus uptake in transgenic tobacco plants that Over-produce citrate[J].Nature Biotechnology,2000:18:450-453
[3] Juan Manuel de la Fueme,Verenice jise,et al.Aluminum tolerance in transgenic plants by alteration of citrate synthesis.Science[J].1997,176:1566-1568
[4] 张丽.烟草柠檬酸合成酶基因对紫花苜蓿的遗传转化[D] :[学位论文].重庆:西南大学,2008
[5] 苏加楷.中国牧草新品种选育的回顾和展望[J].草原与草坪,2001,(4):3-9
[6] 李剑峰.紫花苜蓿在酸铁胁迫下的适应性研究[D] :[学位论文].兰州:甘肃农业大学,2007:34-36
[7] Foy CD.Plant adaptation to acid,aluminum-toxic soils[J].Communications in Soil Science and Plant Analysis.1988,19:959-987
[8] Taylor G J.The physidogy of aluminum tolerance in higher plants[J].Communications in Soil Science and Plant Analysis,1988,19:1179-1194
[9] Alva A K,Edwards D G,Asher C J,et al.Effects of phosphorus/aluminum molar ratio and calcium concentration on plant response to aluminum toxicity[J].Soil Science Society of America Journal,1986,50:133-137
[10] Shen Z G,Wang J L,Guan H Y.Effect of aluminum and calcium on growth of wheat seedlings and germination of seeds[J].Joumal of Plant Nutrition,1993.16:2136-2148
[11] Bessho T,Bell L C.Soil solid and solution phase changes and mung bean response during amelioration of aluminum toxicity with organic mater[J].Plant and Soil,1992,140:181-196
[12] 余国泰,秦遂初.有机肥缓解小麦铝毒效果的研究[J].植物营养与肥料学报,1998,4:57-62
[13] Ahmad F,Tan K H.Effects of lime and organic matter on soybean seedlings grown in aluminum toxic soil[J].Soil Science Society of America Joumal,1986,50:656-661
[14] 沈同,王镜岩,赵邦悌,等.生物化学(第二版)下册[M].北京:高等教育出版社,2000:91-100
[15] Yang Z M,Yang H,Wang J,et al.Aluminum regulation of citrate metabolism for Al-induced citrate efflux in the roots of Cassia tora L[J].Plant and Soil.,2004,166:1589-1594
[16] 李德华,贺立源,刘武定.耐铝的和对铝敏感的玉米自交系根系的有机酸分泌[J].植物生理与分子生物学学报,2003,29(2):114-120
[17] 王建林,曹志洪.根际营养环境与持续农业.植物生理学通讯[J] ,1993,29(5):329-336
[18] Hoffland E,Findenegg G R,Nelemans J A.Solubi lization of rock phosphate by rape.2.Local root exudation of organic acids as a response to P starvation[J].Plant and Soil,1989,113:161-165
[19] Koyama H,Takita E,Kawamura A,et al.Over expression of mitochondrial citrate synthase gene improves the growth of carrot cells in Al-phosphate medium[J].Plant Cell Physiology.1999,40:482-488
[20] 刘小琳,康俊梅,孙彦,等.MsNHXl基因转化紫花苜蓿及转基因植株的鉴定[J].草地学报,2008,16(2):119-120

紫花苜蓿柠檬酸合成酶基因的克隆及对烟草的转化

Cloning of MsCS Gene from Medicago sativa L. and Transformation of Tobaccos

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	王静, 刘晓静, 程甜甜, 童长春, 汪雪. 不同氮效率紫花苜蓿叶特征及其产量效应研究[J]. 草地学报, 2021, 29(9): 1941-1949.
[2]	孟宣辰, 马鹏程, 马杰, 段珍, 韩阳阳, 张吉宇. 黄土高原半干旱区紫花苜蓿覆膜垄沟和施肥效应研究[J]. 草地学报, 2021, 29(9): 2098-2106.
[3]	陈雷, 暴雪艳, 郭刚, 霍文婕, 李清宏, 许庆方, 王聪, 刘强. 单宁酸和乳酸菌对紫花苜蓿青贮品质和体外瘤胃发酵的影响[J]. 草地学报, 2021, 29(8): 1853-1858.
[4]	金一锋, 陈阳, 齐欣, 高岩松, 金忠民, 赵清峰, 王琦. 草地早熟禾NRT1/PTR FAMILY 8.3基因的克隆及表达分析[J]. 草地学报, 2021, 29(7): 1397-1405.
[5]	李春萍, 邱轶辉, 夏厚胤, 谭璐娜, 许环宇, 张瀚文, 高景慧. 紫花苜蓿多父本F₁代低蒸腾及产量性状的隶属函数分析[J]. 草地学报, 2021, 29(7): 1416-1422.
[6]	李天琦, 林志玲, 卢轩, 黄佳媛, 杨宁, 沃野, 高凯, 柳小妮, 王显国. 灌溉量对科尔沁沙地紫花苜蓿土壤速效养分分布及淋失的影响[J]. 草地学报, 2021, 29(7): 1454-1461.
[7]	李鹏珍, 赵得琴, 邓波, 杨军, 赵国华, 赵亮. 生物炭与干旱胁迫对接种紫花苜蓿光合效率及生长的影响[J]. 草地学报, 2021, 29(6): 1257-1264.
[8]	王文静, 麻冬梅, 赵丽娟, 马巧利. 2,4-表油菜素内酯对盐胁迫下紫花苜蓿生理指标及根系离子积累的影响[J]. 草地学报, 2021, 29(6): 1363-1368.
[9]	潘新怡, 史昆, 龚攀, 韦宝, 吴欣明, 王赞. 紫花苜蓿花青苷合成的转录组分析[J]. 草地学报, 2021, 29(5): 866-875.
[10]	黄荣, 姚博, 张玉娟, 贾倩英, 李向阳, 张宏, 张振粉. 成团泛菌CQ10脂多糖对紫花苜蓿种子萌发及苗期生长的影响[J]. 草地学报, 2021, 29(5): 965-971.
[11]	徐洪雨, 甄莉丽, 李钰莹, 董宽虎, 李向林. 低温干旱环境对紫花苜蓿根颈耐寒性的影响[J]. 草地学报, 2021, 29(4): 724-733.
[12]	李满有, 马忠仁, 王斌, 杨雨琦, 沈笑天, 曹立娟, 李小云, 兰剑. 宁夏引黄灌区2个苜蓿品种不同行比混播模式研究[J]. 草地学报, 2021, 29(4): 798-804.
[13]	侯红雁, 申晨, 霍文婕, 刘强, 张拴林, 王聪, 陈雷, 许庆方, 王永新, 王忠豪, 郭刚. 同型发酵乳杆菌对低水分紫花苜蓿青贮发酵品质及体外消化率的影响[J]. 草地学报, 2021, 29(4): 835-841.
[14]	陈德奎, 郭香, 郑明扬, 陈晓阳, 周玮, 张庆. 砂仁精油对紫花苜蓿青贮品质的影响[J]. 草地学报, 2021, 29(4): 855-860.
[15]	夏方山, 王聪聪, 李红玉, 刘曼, 郑川, 张耀丹, 范华芳. 硒引发对紫花苜蓿种子抗氧化性能的影响[J]. 草地学报, 2021, 29(3): 472-477.