马铃薯不同组织的诱导分化及其对遗传转化效率的影响  

Influence on the Conversion Efficiency of Induced Differentiation of Various Potato Tissues

作　　者：朱雪瑞 季静[1] 王罡[1] 马志刚[2] 杨丹[1] 金超[1] 李辰[1] 

机构地区：[1]天津大学环境科学与工程学院,天津300072 [2]天津大学化工学院,天津300072

出　　处：《中国生物工程杂志》2016年第10期53-59,共7页China Biotechnology

基　　金：国家自然科学基金面上项目(2013B1-0043);国家自然科学基金青年项目(31300329)资助项目;国家科技重大专项(2014Z1-0002)

摘　　要：在农业快速发展过程中,基因工程作为改造马铃薯性状的重要手段,一直备受关注。优化马铃薯组织培养体系及外源基因转化条件是进行马铃薯转基因工作的基础。以三种熟性不同的马铃薯栽培品种(东农303、早大白、大西洋)的试管苗为试验材料,采用正交实验,对茎段和试管薯的分化体系进行筛选、优化,建立不同品种茎段的愈伤再生体系及试管薯直接分化再生体系。将以LYCB为目的基因,以NPTII为筛选标记的载体,利用农杆菌转化法,分别对三种材料试管苗的茎段及试管薯进行转基因操作,并对转化条件进行优化,以建立适合于不同品种马铃薯茎段及试管薯的遗传转化体系。以研究中得到的最佳离体再生体系及最优遗传转化体系为基础,利用PCR方法,进行阳性植株检测并统计。研究发现,在东农303、早大白、大西洋三种品种中,经由茎段愈伤组织转化,获得的阳性植株转化率分别为36%、35%、28%;经由试管薯直接分化,获得的阳性植株转化率分别为43%、45%、17%。在后期转基因操作中,东农303/早大白转化时可采用试管薯作为试验组织,大西洋则适合用茎段。With rapid development of agriculture,genetic engineering as an important means of transformation of the potato traits has been extensively applied. Optimization of potato tissue culture system and exogenous gene conversion conditions are the basis of the work of the transgenic potato. Three potato cultivars having different maturity period（ Dongnong 303,Zaodabai,Atlantic） have been tested,with the use of orthogonal experiment,screen,optimize stem segments and Microtuber differentiation system. Stem callus regeneration system and in vitro differentiation of potato direct regeneration system were built. Then applied genetic transformation with Agrobacterium tumefaciens including the reconstruction vector carrying the target gene LYCB and selection marker NPTII. Respectively,the three materials stem section plantlets and microtubers were manipulated,and the transgenic conversion conditions were optimized for each one. Then the three were applyied to the best regeneration system and genetic transformation system,finally positive plants using PCR screening and statistically analysised the data were got. The results show that conversion rate via stem callus of Dongnong 303,Zaodabai and Atlantic respectively were 36%,35%,28%; direct differentiation rate via Microtuber of them separately were43%,45%,17%. In conclusion,microtuber of Dongnong 303 / Zaodabai is suitable for the conversion tissue of test organization,stem of the Atlantic is better conversion one.

关 键 词：马铃薯 试管薯 再生体系 直接分化 遗传转化 

分 类 号：Q812[生物学—生物工程]