苜蓿花药悬浮培养体系的建立及其影响因素

doi:10.11733/j.issn.1007-0435.2010.03.009

›› 2010, Vol. 18 ›› Issue (3): 358-364.DOI: 10.11733/j.issn.1007-0435.2010.03.009

苜蓿花药悬浮培养体系的建立及其影响因素

姚喜红^1,3, 魏臻武^1,2, 耿小丽^1,3

1. 甘肃农业大学草业学院, 甘肃, 兰州, 730070;
2. 扬州大学动物科学与技术学院, 江苏, 扬州, 225009;
3. 草业生态系统教育部重点实验室, 中-美草地畜牧业可持续发展研究中心, 甘肃, 兰州, 730070

收稿日期:2009-11-25 修回日期:2010-03-29 出版日期:2010-06-15 发布日期:2010-06-15
通讯作者: 魏臻武,E-mail:zhenwu_wei@yahoo.com.cn
作者简介:姚喜红(1983- ),女,甘肃天水人,硕士研究生,研究方向为牧草遗传育种E-mail:yaoxihong@yahoo.com.cn
基金资助:
国家“863”计划项目(2008AA10Z149);国家自然科学基金项目(30972136);国家科技支撑计划项目(2008BADB3B10)资助

Establishment of Alfalfa Anther Suspension Culture System and the Influencing Factors

YAO Xi-hong^1,3, WEI Zhen-wu^1,2, GENG Xiao-li^1,3

1 .Pratacultural College, Gansu Agricultural University, Lanzhou, Gansu Province, 730070, China;
2. College of Animal Science& Technique, Yangzhou University, Yangzhou, Jiangsu Province, 225009, China;
3. Key Laboratory of Grassland Ecosystem of Ministry of Education/Sino-U.S.Centers for Grazingland Ecosystem Sustainability, Lanzhou, Gansu Province, 730070, China

Received:2009-11-25 Revised:2010-03-29 Online:2010-06-15 Published:2010-06-15

摘要/Abstract

摘要： 苜蓿(Medicago sativa L.)花药悬浮培养体系的建立可为苜蓿体细胞杂交、遗传转化以及获得苜蓿单倍体,建立苜蓿加倍单倍体群体(DH)提供快捷有效的途径。采用固-液培养基结合的方式建立苜蓿花药悬浮培养体系,通过探索体系建立的基础条件及其影响因素,可建立适宜的苜蓿花药悬浮培养体系。结果表明,苜蓿悬浮培养体系中,基本液体培养基为NB培养基,培养液最佳组合为NB+2,4-D 0.3 mg·L^-1+NAA 0.3 mg·L^-1,蔗糖2%;建立苜蓿花药悬浮细胞系,应选取淡黄色或乳白色,结构疏松,颗粒状胚性愈伤组织为悬浮培养的基础材料;苜蓿花药悬浮细胞系继代周期为6~8 d;苜蓿花药悬浮培养时初始接种量为2g/30mL时悬浮细胞PCV、鲜重以及干重的增殖倍数最高,增殖倍数均超过4倍;植物生长调节剂2,4-D的添加浓度以0.3 mg·L^-1为宜;糖作为组织培养过程中唯一的碳源,在苜蓿细胞悬浮培养体系中应有一定的浓度范围。因此,苜蓿花药悬浮培养中,蔗糖浓度为2%-3%时效果最好。

关键词: 苜蓿, 悬浮培养, 接种量, 2,4-D浓度, 蔗糖浓度

Abstract: Establishment of alfalfa anther suspension system will offer an effective approach for alfalfa somatic hybridization and genetic transformation,acquisition of alfalfa haploid,and creation of double haploid(DH).The objectives of this research were to established alfalfa suspension system and explore the basic condition and influencing factors.Combination of solid and liquid culture mediums was chosen to establish this system.The results show that NB was the basic liquid medium for the establishment of alfalfa suspension cell culture system and the best liquid medium combination was NB+2,4-D 0.3 mg/L+NAA 0.3 mg/L and sugar concentration was 2%.Embryogenic callus characterized with straw yellow or ivory yellow,graininess,loose structure should be chosen as basic material to alfalfa anther suspension cell system and the subculture cycle was 6-8 d.When the initial inoculation dosage was 2 g/30 mL,the proliferation multiples of Packed Cell Volume(PCV),wet weight,and dry weight of suspension cell was the highest and all of them were above 4 times.When establishing alfalfa suspension cell system,the ideal concentration of 2,4-D was 0.3 mg/L.Being the only carbon source in tissue culture,sugar should have a given concentration range during the establishment of alfalfa suspension cell culture system.And in alfalfa anther suspension culture,sugar concentration at 2%~3% had the best effect.

Key words: Alfalfa, Suspension culture, Inoculation dosage, Concentration of 2,4-D, Sugar concentration

中图分类号:

Q813.1

姚喜红, 魏臻武, 耿小丽. 苜蓿花药悬浮培养体系的建立及其影响因素[J]. , 2010, 18(3): 358-364.

YAO Xi-hong, WEI Zhen-wu, GENG Xiao-li. Establishment of Alfalfa Anther Suspension Culture System and the Influencing Factors[J]. , 2010, 18(3): 358-364.

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.1007-0435.2010.03.009

https://manu40.magtech.com.cn/Jweb_cdxb/CN/Y2010/V18/I3/358

参考文献

[1] 魏臻武,符昕,耿小丽,等.苜蓿遗传多样性和亲缘关系的SSR和ISSR分析[J]. 草地学报,2007,15(2):118-123
[2] 雷艳芳,魏臻武,杨占花,等.蒺藜苜蓿种子产量相关性状的遗传分析[J]. 草地学报,2009,17(3):125-128
[3] Guha S,Maheshwari S C.In vitro production of embryos from anthers of Datura[J]. Nature,1964,204:497
[4] 中国科学院上海植物生理研究所细胞室.植物组织与细胞培养[M]. 上海:上海科学技术出版社,1978:171
[5] Muir W H,Hildebrandt A C,Riker A J.The preparation,isolation and growth in culture of single cells from higher plants.Botanical Society of America,1958,45(8):589-597
[6] Steward F C,Marion O M,Smith J.Growth and organized development of culture cells.I.growth and division of freely suspended cells.Botanical Society of America,1958,45(9):693-703
[7] 刘庆昌,米凯霞,鲁迪慧,等.甘薯胚性细胞系悬浮培养的建立[J]. 作物学报,1997,23(1):22-26
[8] 祁新,王权,顾德峰,等.马铃薯悬浮细胞培养[J]. 吉林农业大学学报,1996,18(1) 21-24
[9] 陈志贤,李淑敏,Trolinder L.等.棉花细胞悬浮培养胚胎发生和植株再生某些特性的研究[J]. 中国农学科学,1987,20(5):6-11
[10] 陈克贵,张义正.甘薯细胞悬浮培养的建立及其生长研究[J]. 应用与环境生物学报,1999,5(3):275-278
[11] 孟新亚,邵玉春,刘冬霞,等.大蒜胚性悬浮细胞系的建立[J]. 河南农业科学,2002,6:25-27
[12] 胡张华,陈火庆,吴关庭,等.高羊茅悬浮细胞系的建立及绿色植株的高频再生[J]. 草业学报.2003,6:95-99
[13] Bingham E T,Hurley L V,Kaatz D M.Breeding alfalfa which regenerates from callus tissue in culture.Crop Science,1975,15:719-721
[14] Kao K N,Michayluk M R.Embryoid formation in alfalfa cell suspension cultures from different plants[J]. Tissue Culture Association,1981,7:645-648
[15] Tamas K,Robertson A J,Waterborg J H.Histone H3 transcript stability in alfalfa[J]. Plant Molecular Biology,1995,28:901-914
[16] Jiang Y,Miles P W.Phenolic acids in filtrates of Lucerne suspension cell cultures treated with saliva of the spotted alfalfa aphid,Therioaphis trifolii maculata[J]. Entomologia Experunentalis et Applicata,1996,80:551-553
[17] Qamar S F A,Sors T G,Cunningham S M.Phosphate nutrition effects on growth,phosphate transporter transcript levels and physiology of alfalfa cells[J]. Plant Cell,Tissue and Organ Culture,2005,82:131-140
[18] 张晓东,林延安.苜蓿细胞悬浮培养与耐受高浓度PEG变异体的筛选[J]. 核农学报,1994,8(1):7-13.
[19] 杨占花,魏臻武,张丽芳.两类SSR对苜蓿属种质遗传多样性和亲缘关系的比较研究[J]. 草地学报,2008,16(5):432-438.
[20] 耿小丽,魏臻武,程鹏舞,等.苜蓿花药培养与倍性鉴定[J]. 草原与草坪,2008,130 (5):1-5
[21] McDonald K,Alan J.Bioreactor studies of growth and nutrient utilization in alfalfa suspension cultures.Plant Cell Reports,1989,8:455-458
[22] 谢华永,诸葛强.杨树胚性悬浮细胞系建立与原生质体融合研究[D] :[学位论文]. 南京:南京农业大学,2004,1-48
[23] Gamborg O L,Shyluk J P.Nutrition medium and characteristics of plant cell and tissue cultures.In:Thorpe T A.Plant tissue culture:Methods and Application in Agriculture[M]. New York:Academic press,1981,21-42.
[24] 崔红,陈亮,沈明山,等.甘薯胚性悬浮系的建立及其细胞生长特性研究[J]. 厦门大学学报,2002,41(5):55-58

苜蓿花药悬浮培养体系的建立及其影响因素

Establishment of Alfalfa Anther Suspension Culture System and the Influencing Factors

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]	孙伟, 刘玉玲, 王德平, 赵敏, 杨合龙, 宋倩, 肖红, 王开丽, 曹婧, 戎郁萍. 补播羊草和黄花苜蓿对退化草甸植物群落特征的影响[J]. 草地学报, 2021, 29(8): 1809-1817.
[4]	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.
[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]	李红, 李波, 方志坚, 杨曌, 张超, 冯成龙, 金姗姗, 王思琦, 石婉莹. 冻融和冻融+苏打盐碱复合胁迫下苜蓿幼苗的生理响应[J]. 草地学报, 2021, 29(7): 1469-1476.
[8]	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.
[9]	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.
[10]	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.
[11]	孙雷雷, 赵桂琴, 柴继宽, 刘富渊, 聂秀美, 王军. 贮藏方式和贮藏时间对苜蓿干草霉菌数量和种类的影响[J]. 草地学报, 2021, 29(5): 884-893.
[12]	陈彩锦, 张尚沛, 师尚礼, 曾燕霞, 吴娟, 尚继红, 高婷, 张蓉. 基于GGE双标图对苜蓿品种丰产性和稳定性综合评价[J]. 草地学报, 2021, 29(5): 912-918.
[13]	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.
[14]	陈卫东, 张玉霞, 丛百明, 阿玛尔, 田永雷, 张庆昕, 杜晓艳. 钾肥对紫花苜蓿根颈丙二醛、可溶性蛋白含量与抗氧化系统的影响[J]. 草地学报, 2021, 29(4): 717-723.
[15]	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.