甜瓜再生体系的建立
2021-07-19祁宏英徐洪国王秀文王芳尹帮果高美玲
祁宏英 徐洪国 王秀文 王芳 尹帮果 高美玲



摘 要:以葫芦科的甜瓜为试验对象,探讨不同基因型、不同激素浓度配比及抗生素胁迫对其不定芽萌发的影响,以期建立甜瓜再生培养体系,为甜瓜遗传转化奠定基础。结果表明,甜瓜在初代培养时,不定芽发生的培养基为MS+1.0 mg·L-1 6-BA,甜瓜再生率最高,达到92.5%;甜瓜在继代培养中,0.3 mg·L-1 6-BA的处理最适宜不定芽的伸长生长,0.7 mg·L-1 6-BA的处理甜瓜不定芽增殖系数最高达到2.33;当Kan(卡那霉素)质量浓度大于 20 mg·L-1 时,外植体基本全部褐化,无法分化出绿芽。Amp(氨苄青霉素)在一定质量浓度下对不定芽发生的影响差异性不显著。
关键词:甜瓜;不定芽;再生;抗生素
中图分类号:S652 文献标志码:A 文章编号:1673-2871(2021)05-105-04
Optimization of regeneration system of melon
QI Hongying1,2, XU Hongguo1,2, WANG Xiuwen1, WANG Fang1,2, YIN Bangguo1, GAO Meiling1,2
(1.College of Life Science and Agriculture Forestry, Qiqihar University, Qiqihar 161006, Heilongjiang, China; 2. Heilongjiang Provincial Key Laboratory of Resistance Gene Engineering and Protection of Biodiversity in Cold Areas, Qiqihar 161006, Heilongjiang, China)
Abstract: In this experiment, the effects of different hormone combinations on the regeneration system of different genotypes of melon were studied,and the effects of different hormone combinations and genotypes on the regeneration of melon lines were studied, and the effects of antibiotics on the tolerance of adventitious buds of melon were explored. The aim was to optimize the regeneration culture system of muskmelon and lay the foundation for genetic transformation of melon. The results showed that when the medium of adventitious bud was MS+1.0 mg·L-1 6-BA, the regeneration rate of muskmelon was the highest up to 92.5%. The concentration of 0.3 mg·L-1 6-BA in muskmelon was the most suitable for the growth of adventitious buds. Under the concentration of 0.7 mg·L-1 6-BA, the highest multiplication coefficient of adventitious buds was 2.33. When the concentration of Kan was higher than 20 mg·L-1, the explant was browning and could not differentiate into green buds. There was no significant difference in the effect of certain concentration of Amp on adventitious bud formation.
Key words: Melon; Adventitious buds; Regeneration; Antibiotics
甜瓜(Cucumis melo)是全國乃至全球普遍种植的一种经济作物,果实口感佳、营养丰富、品味独特,是一种广受人们喜爱的食用果品[1]。大多数研究表明,植物的基因型是其器官发生的主要影响因素。随着植物基因工程的发展,通过转基因技术改良甜瓜品质成为新的研究热点,这为提高甜瓜品质和抗逆性开辟了更加安全可靠、快捷可行的途径[2-3]。建立甜瓜高效再生体系是遗传转化的基础,基因型不同,其再生能力也会受到很大的影响,种间、品种之间的再生能力、再生频率差异显著[4-6]。在农杆菌介导的培养转化过程中,一个很重要的环节是抑制农杆菌的生长,防止农杆菌过度增殖对植物组织细胞造成伤害和影响植株的再生,这就需要用抗生素及时有效地抑制,要求使用的抗生素在不伤害或少伤害受体材料的同时能完全抑制农杆菌的生长[7]。……
