PING Lu,GU Defeng,WEI Zhengyi,et al.Screening of High Frequency Regeneration Materials Suitable for Chloroplast Transformation From Three Cultivars of Brassica oleracea var.acephala[J].Northern Horticulture,2016,40(02):92-99.[doi:10.11937/bfyy.201602025]
三种基因型羽衣甘蓝筛选适合叶绿体转化材料
- Title:
- Screening of High Frequency Regeneration Materials Suitable for Chloroplast Transformation From Three Cultivars of Brassica oleracea var.acephala
- Keywords:
- Brassica oleracea var.acephala; adventitious bud induction; chloroplast transformation; selection; high frequency regeneration
- 文献标志码:
- A
- 摘要:
- 以3种不同基因型的羽衣甘蓝品种“皱叶红心”、“白鸥”和“红鸥”为试材，以MS培养基为基础培养基，分别添加不同质量浓度的6-BA、NAA、IBA激素，筛选不同品种真叶叶片不定芽的诱导频率，同时确定不同材料对除草剂双丙氨膦和抗生素壮观霉素的致死临界质量浓度，为羽衣甘蓝的叶绿体转化研究筛选受体材料。结果表明：影响羽衣甘蓝叶片诱导不定芽因素中，激素质量浓度的影响比品种的影响更大；当诱导培养基中的6-BA质量浓度为1.0 mg/L、NAA质量浓度为0.1 mg/L时，诱导效果最佳，是3个品种的最适培养基；按照由高到低的顺序排列，3个品种的不定芽诱导率依次为“红鸥”、“白鸥”和“皱叶红心”，因此择优利用品种“红鸥”作为后续转基因研究的受体材料；分别利用双丙氨膦和壮观霉素的不同质量浓度梯度进行筛选，研究二者对“红鸥”品种叶片分化的影响，发现双丙氨膦和壮观霉素的最低抑制质量浓度分别为4 mg/L和50 mg/L。该研究结果对今后利用叶绿体遗传转化技术改良羽衣甘蓝品种提供了技术支持。
- Abstract:
- Taking three Brassica oleracea var.acephala cultivars with different color，‘Zhouyehongxin’，‘Bai’ou’ and ‘Hong’ou’ as materials，the frequency of adventitious bud induction were studied and so as to provide explants for subsequent chloroplast transformation.The MS medium was served as basic medium added with different concentrations of 6-BA，NAA and IBA，respectively，with the addition of bialaphos sodium and spectinomycin.The results showed that the impact of cultivar on induction of hormone from leaves of Brassica oleracea var.acephala was more visible than that of adventitious buds.The induction frequency of adventitious buds would be highest when the media were added with concentration of 1.0 mg/L 6-BA and 0.1 mg/L NAA for the cultivars of three Brassica oleracea var.acephala and so-called optimal medium for the three cultivars.According to the order from high to low，the ranking of frequencies of adventitious bud induction for three cultivars was ‘Hong’ou’，‘Bai’ou’ and ‘Zhouyehongxin’.Cultivar ‘Hong’ou’，therefore，was considered to be the suitable explant for the genetic transformation.The minimum inhibitory concentration of bialaphos-sodium and spectinomycin in induction medium for ‘Hong’ou’ cultivar was 4 mg/L and 50 mg/L separately.These results provided technical support for chloroplast transformation of Brassica oleracea var.acephala to implement genetic improvement.
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备注/Memo
第一作者简介：平璐（1990-），女，吉林长春人，硕士研究生，研究方向为园林植物种质资源创新。E-mail：1241036296@qq.com. 责任作者：邢少辰(1964-)，男，博士，研究员，研究方向为叶绿体生物反应器。E-mail：xingsc@cjaas.com. 基金项目：吉林省科技发展计划资助项目（20076021；C42070501）。