厚皮甜瓜'SN-1'组培再生和遗传转化体系的建立
Establishment of Tissue Culture and Genetic Transformation Systems for Thick-peeled Melon'SN-1'
赵欣冉 1孙科新 1王建全 1胡宗霞 1郭永杰 1史庆华 2杨晓玉2
作者信息
- 1. 山东农业大学园艺科学与工程学院,山东泰安 271018
- 2. 山东农业大学园艺科学与工程学院,山东泰安 271018;农业农村部黄淮地区园艺作物生物学与种质创制重点实验室,山东 泰安 271018
- 折叠
摘要
本研究以厚皮甜瓜自交系'SN-1'(Cucumis melo ssp.melo)为试材,通过对种子和外植体处理条件、培养基组分、激素配比、共培养条件等进行筛选,建立厚皮甜瓜自交系'SN-1'的组培再生和遗传转化体系.结果表明,种子在Agar培养基的萌发时间显著优于1/2MS+Agar和MS+Agar;分析不同浓度(2%、3%和4%,v/v)NaClO消毒剂和处理时间(10、15、20和25 min)对种子萌发的影响,发现3%NaClO处理15 min的材料萌发率和整齐度均显著优于其他处理;相较"二分法","四分法"在不影响分化的前提下,可获得更多子叶节、显著提高种子的利用率;进一步分析发现,以播种后48 h、胚根长约5 mm的种子收集子叶节,不定芽分化效果显著优于其他时期;研究不同激素配比和浓度对子叶节的影响,发现培养基中6-BA、IAA和ABA浓度分别为0.5 mg·L-1、0.05 mg·L-1和1 mg·L-1时,不定芽分化最佳;当生根培养基中添加0.4 mg·L-1IAA时,再生苗不定根诱导效果显著优于其他处理;对侵染后的共培养周期和培养基激素配比进行分析,发现当共培养3 d、且培养基中添加0.5 mg·L-1 6-BA+0.1 mg·L-1 IAA时,子叶节发育最佳.
Abstract
We used the self-pollinated line'SN-1'(Cucumis melo ssp.melo)of muskmelon as the experimental material.By screening conditions for seed and explant treatment,culture medium components,hormone ratios,and co-culture conditions,a tissue culture regeneration and genetic transformation system for the self-pollinated line'SN-1'of muskmelon was established.The results indicated that the germination time of seeds on Agar medium was significantly better than on 1/2MS+Agar and MS+Agar;analyzing the effects of different concentrations(2%,3%,and 4%,v/v)of NaClO disinfectant and treatment times(10,15,20,and 25 min)on seed germination revealed that treatment with 3%NaClO for 15 minutes resulted in significantly higher germination rates and uniformity compared to other treatments;compared to the"binary method,"the"quaternary method"can obtain more cotyledonary nodes without affecting differentiation,significantly improving seed utilization;further analysis found that collecting cotyledonary nodes from seeds with a radicle length of about 5 mm at 48 hours after sowing resulted in significantly better adventitious bud differentiation than other periods;research on the effects of different hormone ratios and concentrations on cotyledonary nodes revealed that the best adventitious bud differentiation was achieved with concentrations of 0.5 mg·L-1 6-BA,0.05 mg·L-1 IAA,and 1 mg·L-1 ABA in the culture medium;when adding 0.4 mg·L-1 IAA to the rooting medium,the induction effect of adventitious roots in the regenerated seedlings was significantly better than other treatments;analysis of the co-culture period and medium hormone ratios after infection revealed that the best cotyledonary node development occurred with a 3-day co-culture period and the addition of 0.5 mg·L-1 6-BA+0.1 mg·L-1 IAA to the medium.
关键词
厚皮甜瓜/组培再生/遗传转化Key words
Cucumis melo L.ssp.melo/tissue culture/genetic transformation引用本文复制引用
基金项目
国家自然科学基金面上项目(32272795)
山东省泰山学者青年专家项目(tsqnz20230605)
山东省自然科学基金面上项目(ZR2022MC029)
山东省农业良种工程项目(2022LZGCQY006)
山东省农业良种工程项目(2023LZGCQY021)
山东省蔬菜产业技术体系项目(SDAIT-05)
出版年
2024
国家科技期刊平台
NSTL
万方数据