文档介绍：农杆菌介导的玉米遗传转化及再生体系的建立
摘要: 为了建立玉米高频再生及高效遗传转化体系,本研究选用玉米自交系178,昌72,郑58,H736760A,MQ704,2193为材料,对影响玉米成熟胚愈伤组织诱导、继代培养及分化的因素进行了研究。结果表明:高浓度的2,4-二氯苯氧乙酸(2,4-D)()是诱导愈伤组织必须的;在继代培养基中添加适量的2,4-D()、6-苄基嘌呤(6-BA)()和硝酸银(AgNO3)(10mg/L)能显著增加胚性愈伤组织的形成; 6-BA有利于提高愈伤组织的分化频率。这个体系的建立,为以成熟胚为受体系统的遗传转化体系奠定了基础。
关键词: 玉米;成熟胚;愈伤组织;转化;植株再生
Establishment of transformation system and regeneration system
of maize mediated by Agrobacterium tumefaciens
Abstract: In order to establish the system of high-frequency regeneration and high-efficiency ic transformation in maize, the inbred line 178, Chang72, Zheng58, H736760A, MQ704, 2193 have been chosen to study on initiation, maintenance and differentiation of calli in this paper. The results showed that inclusion higher concentration of 2,4-Dichlorophenoxyacetic acid(2,4-D, mg/L) into induction medium was a critical factor for producing primary calli. Supplement with optimal level of 2,4-D( mg/L), 6-Benzylaminopurine(6-BA, mg/L)and silver nitrate(10 mg/L)in the subculture medium significantly promoted the formation of embryogenic calli. And the addition mg/L of 6-BA into regeneration medium has significantly increased the frequency of plant regeneration. This efficient regeneration system provides a solid basis for ic transformation of maize.
Key words: Maize; Mature embryo; Calli; Transformation; Plant regeneration
摘要
ABSTRACT
目录
1序言 4
2玉米再生及基因转化体系的研究 4
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电激法 6
花粉管通道法 7
PEG法 7
基因枪法 7
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3玉米成熟胚再生体系的建立 11
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材料 11
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外植体的选择与预培养 11
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胚性愈伤组织的形成及愈伤组织的继代 12
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胚性愈伤组织的形成 15
AgNO3对胚性愈伤组织形成的影响 16
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结论与讨论 17
4 农杆菌介导的玉米转化体系的建立 18