利用胀果甘草无菌苗根作为外植体诱导愈伤组织,经过多次继代培养筛选出4种类型的愈伤组织,并以不同质量浓度的NaCl胁迫处理Ⅰ型愈伤组织.测定了在根愈伤组织诱导过程中,不同类型愈伤组织继代培养中和Ⅰ型愈伤组织在NaCl胁迫处理条件下的生长情况及可溶性蛋白含量的变化情况.结果显示,以苗龄10 d的无菌苗根为外植体可以诱导产生愈伤组织,以MS培养基附加1.0 mg/L 2,4-D和 0.2 mg/L 6-BA时,根愈伤组织的诱导率最高,继代培养生长状况良好为Ⅰ型愈伤组织;根外植体两端开始形成愈伤组织时可溶性蛋白含量最高,为2.01 mg/g;不同培养基上继代培养的根愈伤组织生长状态不同,可溶性蛋白含量也有不同,以Ⅰ型愈伤组织中可溶性蛋白含量最高;Ⅰ型愈伤组织在5 g/L NaCl胁迫下,继代培养21 d后,愈伤组织生长量明显提升,在14~21 d愈伤组织可溶性蛋白含量明显提高.本研究表明甘草愈伤组织对低质量浓度的盐胁迫有一定的适应性,使其可溶性蛋白含量升高,在可溶性蛋白含量剧烈变化期间,愈伤组织生长量明显提高.
The root of Glycyrrhiza inflata aseptic seedling was used as explant to induce callus in this study. Four types of callus were selected after several rounds of subculture and type I callus selected was treated by NaCl stress of different mass concentrations. The growth and the amounts of soluble proteins in the root callus during the period of induction, in the callus subcultured on different media and in typeⅠcallus treated with NaCl stress were measured. The results show that the callus could be induced using the root of 10 d aseptic seedling as explant. The highest induction rate of root callus was obtained by the addition of 1.0 mg/L 2, 4-D and 0.2 mg/L 6-BA in MS medium. The highest soluble protein content of callus was 2.01 mg / g at initiation of the induction from the two ends of the root explants. Both the growth of- DOI:10.3969/j.issn.1000-1565.2018.03.010甘草根愈伤组织诱导与可溶性蛋白含量变化马健1,梁玉玲1,2,潘笑1,刘静1(1.河北大学 生命科学学院,河北 保定 071002;2.河北省生物工程技术研究中心,河北 保定 071002)摘 要:利用胀果甘草无菌苗根作为外植体诱导愈伤组织,经过多次继代培养筛选出4种类型的愈伤组织,并以不同质量浓度的NaCl胁迫处理Ⅰ型愈伤组织.测定了在根愈伤组织诱导过程中,不同类型愈伤组织继代培养中和Ⅰ型愈伤组织在NaCl胁迫处理条件下的生长情况及可溶性蛋白含量的变化情况.结果显示,以苗龄10 d的无菌苗根为外植体可以诱导产生愈伤组织,以MS培养基附加1.0 mg/L 2,4-D和 0.2 mg/L 6-BA时,根愈伤组织的诱导率最高,继代培养生长状况良好为Ⅰ型愈伤组织;根外植体两端开始形成愈伤组织时可溶性蛋白含量最高,为2.01 mg/g;不同培养基上继代培养的根愈伤组织生长状态不同,可溶性蛋白含量也有不同,以Ⅰ型愈伤组织中可溶性蛋白含量最高;Ⅰ型愈伤组织在5 g/L NaCl胁迫下,继代培养21 d后,愈伤组织生长量明显提升,在14~21 d愈伤组织可溶性蛋白含量明显提高.本研究表明甘草愈伤组织对低质量浓度的盐胁迫有一定的适应性,使其可溶性蛋白含量升高,在可溶性蛋白含量剧烈变化期间,愈伤组织生长量明显提高.关键词:胀果甘草;根;愈伤组织;盐胁迫;可溶性蛋白中图分类号:Q943.1 文献标志码:A 文章编号:1000-1565(2018)03-0291-08Induction of callus from Glycyrrhiza inflata root andchanges of soluble protein contentMA Jian1,LIANG Yuling1,2,PAN Xiao1,LIU Jing1(1.College of Life Sciences,Hebei University,Baoding 071002,China;2.Biological Engineering Technology Research Center of Hebei Province, Baoding 071002,China)Abstract:The root of Glycyrrhiza inflata aseptic seedling was used as explant to induce callus in this study. Four types of callus were selected after several rounds of subculture and type I callus selected was treated by NaCl stress of different mass concentrations. The growth and the amounts of soluble proteins in the root callus during the period of induction, in the callus subcultured on different media and in typeⅠcallus treated with NaCl stress were measured. The results show that the callus could be induced using the root of 10 d aseptic seedling as explant. The highest induction rate of root callus was obtained by the addition of 1.0 mg/L 2, 4-D and 0.2 mg/L 6-BA in MS medium. The highest soluble protein content of callus was 2.01 mg / g at initiation of the induction from the two ends of the root explants. Both the growth of- 收稿日期:2017-09-20 基金项目:河北大学自然科学研究计划项目(799207217054);河北省生物学重点学科资助项目 第一作者:马健(1991—),男,山东济南人,河北大学在读硕士研究生,主要从事植物基因与细胞工程研究.E-mail:1140299778@qq.com 通信作者:梁玉玲(1965—),女,河北定州人,河北大学教授,博士,主要从事植物基因与细胞工程研究.E-mail:yuling_liang@163.com第3期马健等:甘草根愈伤组织诱导与可溶性蛋白含量变化callus subcultured on different media and the contents of soluble proteins were found to be different with the greatest change found in type I callus. The growth of typeⅠcallus was improved significantly after subculture for 21 d under 5 g/L NaCl;and the soluble proteins of the callus in 14~21 d was dramatically increased. The present study shows that Glycyrrhiza inflata licorice root-induced callus has some adaptability to NaCl stress at the concentration of 5 g/L, and the biggest change of callus growth was accompanied by the significant variation of soluble proteins.
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