|
|
- 2018
以诺丽茎段为外植体的无籽基因转化研究
|
Abstract:
以诺丽无菌苗不带腋芽的茎段为转化材料，采用根癌农杆菌介导法，将含有无籽基因的质粒pCAMBIA1305.1导入到诺丽细胞中，通过Kan快速筛选、PCR检测等方法对转化子进行鉴定.实验结果表明：抗性芽的平均分化频率为42%，经过PCR检测，以茎为外植体进行遗传转化的转化率为5.4%.在转化过程中，有些植株可能会发生饰变.该实验初步证实已把无籽基因转入诺丽，后续需Southern杂交进一步验证.
In this study, the method of Agrobacterium-mediation was used to import pCAMBIA 1305.1, which carries the seedlessness gene, into the cells of the stem segments without axillary buds of aseptic noni (Morinda citrifolia). Then we identified the transformant with the methods of Kan quick screening and PCR checking. PCR results showed that the average differentiation rate of resistant buds was 42%, and the conversion rate of the genetic transformation using noni stem segments as the explants was 5.4%. Furthermore, in the process of transformation, modification could occur in some plants. This study preliminarily confirmed that the seedlessness gene had been transformed into the noni plants, but Southern hybridization is to be made in further study to verify the results of this study
| [1] | 李奕星, 袁德宝, 郑晓燕, 等. 诺丽果汁的抗氧化性研究[J]. 热带作物学报, 2013, 34(8): 1531-1534. DOI:10.3969/j.issn.1000-2561.2013.08.022 |
| [2] | 沈烨.根癌农杆菌介导的甜樱桃茎尖遗传转化体系的建立[D].重庆: 西南农业大学, 2002. http://www.wanfangdata.com.cn/details/detail.do?_type=degree&id=Y458549 |
| [3] | GIRI C C, SHYARNKUMAR B, ANJANEYULU C. Progress in Tissue Culture, Genetic Transformation and Applications of Biotechnology to Trees:All Overview[J]. Trees Struct Funcu, 2004, 18(2): 115-135. DOI:10.1007/s00468-003-0287-6 |
| [4] | 李菊, 程春振, 张永艳, 等. 柑橘和拟南芥RIN4基因的过表达载体构建及其对沙田柚的DNA转化[J]. 西南大学学报(自然科学版), 2015, 37(3): 16-21. |
| [5] | 张海芹, 刘雅丽, 张锋, 等. 根癌农杆菌介导的葡萄风信子遗传转化体系[J]. 西北农林科技大学学报(自然科学版), 2017, 45(2): 135-142. |
| [6] | 于波, 吴华玲. 根癌农杆菌介导的巴西橡胶树胚性悬浮细胞的遗传转化[J]. 中国农学通报, 2012, 28(31): 15-20. DOI:10.3969/j.issn.1000-6850.2012.31.003 |
| [7] | 潘晓晴, 吴田, 蓝增全, 等. 诺丽茎段的离体再生[J]. 东北林业大学学报, 2014, 42(8): 20-24. DOI:10.3969/j.issn.1000-5382.2014.08.005 |
| [8] | 中国科学院植物研究所. 中国高等植物图鉴:第4册[M]. 北京: 科学出版社, 1975: 135-142. |
| [9] | 徐春香, 何勇强, 尉义明, 等. 抗冻蛋白(afp)基因表达载体的构建及对香蕉胚性细胞悬浮系的转化[J]. 广西植物, 2009, 29(5): 664-668. DOI:10.3969/j.issn.1000-3142.2009.05.021 |
| [10] | 吴田, 蓝增全. 诺丽离体培养研究[J]. 山东农业大学学报(自然科学版), 2011, 42(2): 179-182. DOI:10.3969/j.issn.1000-2324.2011.02.003 |
| [11] | 黄骐, 何文锦, 叶冰莹, 等. 诺丽离体快速繁殖研究[J]. 福建师范大学学报(自然科学版), 2007, 23(1): 87-90. DOI:10.3969/j.issn.1000-5277.2007.01.021 |
| [12] | 甘炳春, 何明军. 海巴戟天的栽培及其利用[J]. 中国林副特产, 2004, 2(69): 7-8. |
| [13] | 孙春玉, 孙玚, 刘庆忠, 等. 根癌农杆菌介导的苹果遗传转化研究进展[J]. 中国农学通报, 2010, 26(4): 231-233. |
| [14] | 董志刚, 马小河, 王国平, 等. 根癌农杆菌介导的葡萄遗传转化体系的优化[J]. 核农学报, 2014, 28(7): 1169-1174. |
| [15] | LINDSEY K, CALLOIS P. Transformation of Sugarbeet (Beta vulgaris L.) by Agrobacterium Tumefaciens[J]. Exp Bot, 1990, 41: 529-536. DOI:10.1093/jxb/41.5.529 |
| [16] | PEYMAN NOROUZI, MOHAMMAD A M, KATAYOUN ZAMANI, et al. Using a Competent Tissure for Efficent Transformation of sugarbeet (Beta vulgaris L.) in Vitro Cell[J]. Dev Biol Plant, 2005, 41: 11-16. DOI:10.1079/IVP2004589 |
| [17] | 江一北, 李政, 林晓露, 等. 农杆菌介导的美洲商路抗病毒蛋白基因对非洲菊遗传转化研究[J]. 西南师范大学学报(自然科学版), 2010, 35(3): 76-80. |
| [18] | ROUX N S, TOLOZA A, DOLEZEL J, et al. Usefulness of Embrogenic Cell Suspension Cultures for the Induction and Selection of Mutants in Musa spp[C]. Jain S M, Swennen R. Banana Improvment: Cellular, Molecular Biology, and Induced Mutations. Enfield USA: Science Publishers, 2004, Inc: 33-43. |
| [19] | 熊换英, 钟伟光, 张寿文, 等. 农杆菌介导的植物遗传转化影响因素研究进展[J]. 安徽农业科学, 2012, 40(17): 9214-9217, 9287. DOI:10.3969/j.issn.0517-6611.2012.17.011 |
| [20] | 李玉平, 龚宁, 王永宏, 等. 大花金挖耳细胞悬浮系的建立及黄酮类化合物的产生[J]. 食品科学, 2010, 31(21): 239-243. |
