中国水仙花器官愈伤组织诱导、分化及组织学观察
Callus Induction, Differentiation and Histological Observation of Narcissus tazetta var. chinensis Floral Organs
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摘要: 为了建立中国水仙遗传转化体系,为转基因分子育种提供良好受体,以中国水仙‘金盏银台’品种的不同花器官为实验材料,以MS为基本培养基,研究了花药、花梗、子房、花柄愈伤组织诱导情况。对愈伤组织诱导效果较好的花梗和花药培养基中添加不同浓度6-苄基腺嘌呤(6-BA)、硫酸腺嘌呤(Ad)、萘乙酸(NAA)和2,4-二氯苯氧乙酸(2,4-D),筛选出愈伤组织诱导及鳞茎芽再生的最适培养基配方。结果表明:花梗愈伤组织诱导及鳞茎芽分化的最佳培养基为MS+1 g·L-1 NaH2PO4+0.003 g·L-1 6-BA + 0.001 g·L-1 NAA+0.2 g·L-1 Ad+30 g·L-1 蔗糖,花药愈伤组织诱导及鳞茎芽分化的最佳培养基为MS+0.002 g·L-1 6-BA+0.001 g·L-1 NAA+30 g·L-1 蔗糖;花梗愈伤组织诱导到鳞茎芽分化的时间需要30 35 d,而花药这一过程为80 90 d。对花梗愈伤组织诱导及鳞茎芽分化过程进行组织学观察表明:本研究中产生的小鳞茎是由愈伤组织再分化得来的,表明花梗更有利于作为转基因受体,从而解决了转基因研究过程的一个瓶颈问题。Abstract: In order to select the high-efficient in vitro regeneration approaches for transgenic receptor system, the floral organs of Narcissus tazetta var. chinensis ‘Jinzhanyintai’ variety was served as explants and cultured in the MS basic medium. The influence of plant growth regulators on explants type, such as anther, peduncle, ovary and pedicle were studied. The effects of different concentrations of 6-BA, Ad, NAA, 2,4-D and their combinations on callus induction and shoots regeneration from anthers and peduncles were tested by in vitro culture. The results showed that the suitable media for callus induction and shoots regeneration from peduncles were MS + 30 g·L-1 sucrose with 1 g·L-1 NaH2PO4+0.003 g·L-1 6-BA+0.001 g·L-1 NAA+0.2 g·L-1 Ad, and that from anthers were MS+30 g·L-1 with 0.002 g·L-1 6-BA+0.001 g·L-1 NAA. The process of callus from peduncles and differentiation lasted for 30 35 days and the process of callus from anthers and differentiation lasted for 80 90 days. Shoots were regenerated by peduncles callus for histological examination in the process of callus from peduncles and differentiation.
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Key words:
- Narcissus tazetta var. chinensis
- / callus
- / histological observation
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