欢迎访问作物学报,今天是

作物学报 ›› 2008, Vol. 34 ›› Issue (03): 423-428.doi: 10.3724/SP.J.1006.2008.00423

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

玉米成熟胚胚性愈伤组织的诱导、高频再生及转化的研究

王世玉1; 郑用琏1; 刘亚2; 赵久然2; 张方东1,*

  

  1. 1华中农业大学作物遗传改良国家重点实验室, 湖北武汉130070; 2北京市农林科学院玉米研究中心, 北京100089
  • 收稿日期:2007-06-04 修回日期:1900-01-01 出版日期:2008-03-12 网络出版日期:2008-03-12
  • 通讯作者: 张方东

Transformation, Inducing and High-Frequency Regeneration of Embryogenic Callus Initiated from Mature Embryos of Maize (Zea mays L.)

WANG Shi-Yu1,ZHENG Yong-Lian1,LIU Ya2,ZHAO Jiu-Ran2,ZHANG Fang-Dong1*   

  1. 1 National Key Laboratory for Genetic Improvement of Crops, Huazhong Agricultural University, Wuhan 430070, Hubei; 2 Maize Research Center, Beijing Academy of Agricultural and Forestry Sciences, Beijing 100089, China
  • Received:2007-06-04 Revised:1900-01-01 Published:2008-03-12 Published online:2008-03-12
  • Contact: ZHANG Fang-Dong

摘要: 以玉米自交系CML295、CML304和18-599R的成熟胚为外植体, 结合幼胚离体培养方法, 探讨并优化了成熟胚来源的胚性愈伤组织诱导及继代培养方法。对其愈伤组织的形态和组织切片的研究结果显示, 继代过程可产生良好的Ⅱ型胚性愈伤组织。在分化培养中分别获得68.6%、75.4%和84.8%的高频再生率, 每愈伤组织块成苗数分别为2.45、2.43和2.75。利用基因枪法转化pCAMBIA1301质粒后的GUS瞬时表达效率分别为57.9%、62.5%和73.1%, 转化pCAMBIA1303质粒后检测GFP的瞬时表达效率分别为23.3%、40%和45.5%。以上3种基因型成熟胚来源的愈伤组织转化率与其对应的幼胚来源的胚性愈伤组织转化率相似。这一技术体系为玉米的遗传改良和功能基因组研究提供了重要的技术平台。

关键词: 玉米, 成熟胚, 胚性愈伤组织, 再生, 转化

Abstract: Immature embryo-derived callus is more efficient for transgenic and plant regeneration than calli from other explant tissues. It’s reported that embryogenic callus can be initiated from mature embryos. The use of mature embryos from dry seed are easy to handle, available year round and in bulk quantities. In this study we established an efficient transgenic acceptor system and developed some new methods on maize (Zea mays L.) tissue culture, plant regeneration and genetic transformation with embryogenic callus initiated from mature embryos of three elite inbred lines CML295, CML304, and 18-599R. The calli were cultured in subculture medium of immature embryos. Tissue slice showed that the structure of callus formed from mature embryos was the same as those from immature embryos, being the typeⅡ embryogenic callus. The calli were transferred into regeneration medium of immature embryos supplemented with 2 mg L-1 6-BA. The regeneration frequencies of the calli from mature embryos of CML295, CML304, and 18-599R were 68.6%, 75.4%, and 84.8%, respectively. Vector pCAMBIA1301 was transformed into callus from mature embryos of CML295, CML304, and 18-599R by particle gun using GUS as a report gene; vector pCAMBIA1303 was transformed by particle gun using GFP as a report gene, with GUS staining frequency of 57.9%, 62.5%, and 73.1%; and GFP transient expression frequency of 23.3%, 40%, and 45.5%, respectively. The transgenic rates of embryogenic callus from mature embryos of the three inbred lines were similar to those from immature embryos. In conclusion, using embryogenic callus from mature embryos as transgenic acceptors is efficient and available.

Key words: Mature embryo, Maize, Embryogenic callus, Regenetion, Transformation

[1] 肖颖妮, 于永涛, 谢利华, 祁喜涛, 李春艳, 文天祥, 李高科, 胡建广. 基于SNP标记揭示中国鲜食玉米品种的遗传多样性[J]. 作物学报, 2022, 48(6): 1301-1311.
[2] 崔连花, 詹为民, 杨陆浩, 王少瓷, 马文奇, 姜良良, 张艳培, 杨建平, 杨青华. 2个玉米ZmCOP1基因的克隆及其转录丰度对不同光质处理的响应[J]. 作物学报, 2022, 48(6): 1312-1324.
[3] 王丹, 周宝元, 马玮, 葛均筑, 丁在松, 李从锋, 赵明. 长江中游双季玉米种植模式周年气候资源分配与利用特征[J]. 作物学报, 2022, 48(6): 1437-1450.
[4] 杨欢, 周颖, 陈平, 杜青, 郑本川, 蒲甜, 温晶, 杨文钰, 雍太文. 玉米-豆科作物带状间套作对养分吸收利用及产量优势的影响[J]. 作物学报, 2022, 48(6): 1476-1487.
[5] 陈静, 任佰朝, 赵斌, 刘鹏, 张吉旺. 叶面喷施甜菜碱对不同播期夏玉米产量形成及抗氧化能力的调控[J]. 作物学报, 2022, 48(6): 1502-1515.
[6] 徐田军, 张勇, 赵久然, 王荣焕, 吕天放, 刘月娥, 蔡万涛, 刘宏伟, 陈传永, 王元东. 宜机收籽粒玉米品种冠层结构、光合及灌浆脱水特性[J]. 作物学报, 2022, 48(6): 1526-1536.
[7] 单露英, 李俊, 李亮, 张丽, 王颢潜, 高佳琪, 吴刚, 武玉花, 张秀杰. 转基因玉米NK603基体标准物质研制[J]. 作物学报, 2022, 48(5): 1059-1070.
[8] 许静, 高景阳, 李程成, 宋云霞, 董朝沛, 王昭, 李云梦, 栾一凡, 陈甲法, 周子键, 吴建宇. 过表达ZmCIPKHT基因增强植物耐热性[J]. 作物学报, 2022, 48(4): 851-859.
[9] 刘磊, 詹为民, 丁武思, 刘通, 崔连花, 姜良良, 张艳培, 杨建平. 玉米矮化突变体gad39的遗传分析与分子鉴定[J]. 作物学报, 2022, 48(4): 886-895.
[10] 冯亚, 朱熙, 罗红玉, 李世贵, 张宁, 司怀军. 马铃薯StMAPK4响应低温胁迫的功能解析[J]. 作物学报, 2022, 48(4): 896-907.
[11] 闫宇婷, 宋秋来, 闫超, 刘爽, 张宇辉, 田静芬, 邓钰璇, 马春梅. 连作秸秆还田下玉米氮素积累与氮肥替代效应研究[J]. 作物学报, 2022, 48(4): 962-974.
[12] 徐宁坤, 李冰, 陈晓艳, 魏亚康, 刘子龙, 薛永康, 陈洪宇, 王桂凤. 一个新的玉米Bt2基因突变体的遗传分析和分子鉴定[J]. 作物学报, 2022, 48(3): 572-579.
[13] 宋仕勤, 杨清龙, 王丹, 吕艳杰, 徐文华, 魏雯雯, 刘小丹, 姚凡云, 曹玉军, 王永军, 王立春. 东北主推玉米品种种子形态及贮藏物质与萌发期耐冷性的关系[J]. 作物学报, 2022, 48(3): 726-738.
[14] 渠建洲, 冯文豪, 张兴华, 徐淑兔, 薛吉全. 基于全基因组关联分析解析玉米籽粒大小的遗传结构[J]. 作物学报, 2022, 48(2): 304-319.
[15] 张倩, 韩本高, 张博, 盛开, 李岚涛, 王宜伦. 控失尿素减施及不同配比对夏玉米产量及氮肥效率的影响[J]. 作物学报, 2022, 48(1): 180-192.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
No Suggested Reading articles found!