Induction of callus from Glycyrrhiza inflata root and changes of soluble protein content

doi:10.3969/j.issn.1000-1565.2018.03.010

Abstract

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- 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 Yuling^1,2,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.

Key words: Glycyrrhiza inflata Batal, root, callus, salt stress, soluble protein

CLC Number:

Q943.1

MA Jian,LIANG Yuling,PAN Xiao,LIU Jing. Induction of callus from Glycyrrhiza inflata root and changes of soluble protein content[J]. Journal of Hebei University (Natural Science Edition), 2018, 38(3): 291-298.

References

[1] 许秋霞,邹敏.甘草的药理作用概述[J].实用中医药杂志,2005,21(7):450-451.DOI:10.3969/j.issn.1004-2814.20 05.07.075. XU Q X,ZHOU M.Outlines of pharmacological action of Glycyrrhizae [J].Progress in Modern Biomedicine, 2005,21(7):450-451.DOI:10.3969/j.issn.1004-2814.2005.07.075.
[2] 程焕欣.胁迫条件对胀果甘草愈伤组织及其黄酮含量的影响[D].保定:河北大学,2010.DOI:10.7666/d.d079358. CHENG H X.Effect of abioticstresson growth and liquorice flavonoid production of Glycyrrhiza inflata callus[D]. Baoding:Hebei University,2010.DOI:10.7666/d.d079358.
[3] 谢虹,杨兰,李忠光.脯氨酸在植物非生物胁迫耐性形成中的作用[J].生物技术通报,2011(2):23-27,60.D OI:10.13560/j.cnki.biotech.bull.1985.2011.02.015. XIE H,YANG L,LI Z G.The roles of proline in the formation of plant tolerance to abiotic stress[J].Biotechnology Bulletin,2011(2):23-27,60.DOI:10.13560/j.cnki.biotech.bull.1985.2011.02.015.
[4] 全先庆,张渝洁,单雷,等.高等植物脯氨酸代谢研究进展[J].生物技术通报,2007(1):14-18.DOI:10.3969/j.issn. 1002-5464.2007.01.004. QUAN X Q,ZHANG Y J,SHAN L,et al.Advances in proline metabolism researches of higher plants[J].Bio technology Bulletin,2007(1):14-18.DOI:10.3969/j.issn.1002-5464.2007.01.004.
[5] 董秋丽,夏方山,董宽虎.碱性盐胁迫对芨芨草苗期脯氨酸和可溶性蛋白含量的影响[J].畜牧与饲料科学,2010, 31(4):11-12,28.DOI:10.3969/j.issn.1672-5190.2010.04.005. DONG Q L,XIA F S,DONG K H.Effects of alkaline salinity stress on proline content and soluble protein conte nt of Achnatherum splendens at seedling stage[J].Animal Husbandry And Feed Science,2010,31(4):11-12,28.DOI:10.3969/j.issn.1672-5190.2010.04.005.
[6] 梁玉玲,管延英,阳丽,等.胀果甘草愈伤组织培养及甘草酸含量分析[J].河北大学学报(自然科学版),2000,20 (4):365-368.DOI:10.3969/j.issn.1000-1565.2000.04.016. LIANG Y L,GUAN Y Y,YANG L,et al.Cultivation of callus tissue of Glycyrrhiza inflata and the Glycyrrhizic acid content analysis[J].Journal of Hebei University(Natural Science Edition),2000,20(4):365-368.DOI: 10.3969/j.issn.1000-1565.2000.04.016.
[7] 张坚,高文远,吕连媛,等.甘草不定根培养的研究[J].天津中医药,2011,28(1):75-77. ZHANG J,GAO W Y,LÜ L Y,et al.Studies on culture of adventitious roots of Liquorice[J].Tianjin Journal of Traditional Chinese Medicine,2011,28(1):75-77.
[8] 邓丽莉,潘晓倩,生吉萍,等.考马斯亮蓝法测定苹果组织微量可溶性蛋白含量的条件优化[J].食品科学,2012,3 3(24):185-189. DENG L L,PAN X Q,SHENG J P,et al.Optimization of experimental conditions for the determnation of water soluble protein in Apple pulp using coomassie brilliant blue method[J].Food Science,2012,33(24):185-189.
[9] 张淑娟,王进茂,张世红,等.欧洲白桦叶片器官发生过程中生理指标的变化[J].河北林果研究,2006,21(3):22 9-232.DOI:10.3969/j.issn.1007-4961.2006.03.001. ZHANG S J,WANG J M,ZHANG S H,et al.Study on several physiological indices in the organogenesis of curly birch(Betula pendula ×B. pendula var.carelica Sok.)[J].Hebei Journal of Forestry and Orchard Research,20 06,21(3):229-232.DOI:10.3969/j.issn.1007-4961.2006.03.001.
[10] 李永德,李旭,金香花,等.NaCl胁迫对文冠果幼苗生长和生理生化特征的影响[J].延边大学农学学报,2015,37 (3):212-216.DOI:10.13478/j.cnki.jasyu.2015.03.006. LI Y D,LI X,JIN X H,et al.Growth and physiological characteristics of Xanthoceras sorbifolia seedlings und er NaCl stress[J].Journal of Agricultural Science Yanbian University,2015,37(3):212-216.DOI:10.13478/j.cnki.j asyu.2015.03.006.
[11] 刘亚菊,殷红,朱四易.噻二唑苯基脲在平贝母脱分化过程中引起的生理生化变化[J].西北植物学报,2003,23(3):433-437.DOI:10.3321/j.issn:1000-4025.2003.03.013. LIU Y J,YIN H,ZHU S Y.Thidiazuron induced physiological and biochemical changes in the process of dediffer entiation of Fritillaria ussuriensis[J].Acta Botanica Boreali-occidentalia Sinica,2003,23(3):433-437.DOI:10.3321/ j.issn:1000-4025.2003.03.013.
[12] 张超强,郑秀芳.披针叶黄华试管苗适应盐胁迫的渗透调节机制[J].西北植物学报,2014,34(5):963-969.DOI:1 0.7606/j.issn.1000-4025.2014.05.0963. ZHANG C Q,ZHENG X F.Mechanisms of osmotic adjustment of Themopsis lanceolata test-tuble plantlets in ad apting to salt stress[J].Acta Botanica Boreali-Occidentalia Sinica,2014,34(5):963-969.DOI:10.7606/j.issn.1000- 4025.2014.05.0963.
[13] 杨国会,石德成.甘草对NaCl和Na₂CO₃胁迫的生理响应特点[J].湖北农业科学,2009,48(5):1189-1191,1197.D OI:10.3969/j.issn.0439-8114.2009.05.052. YANG G H,SHI D C. Physiological responses of Glycyrrhiza uralensis Fisch to NaCl and Na₂CO₃ stresses[J].Hubei Agricultural Sciences,2009,48(5):1189-1191,1197.DOI:10.3969/j.issn.0439-8114.2009.05.052.
[14] 汪贵斌,曹福亮.盐胁迫对落羽杉生理及生长的影响[J].南京林业大学学报(自然科学版),2003,27(3):11-14.D OI:10.3969/j.issn.1000-2006.2003.03.003. WANG G B,CAO F L.Effects of salinity on growth and physiology of Bald Cypress Seedlings[J].Journal of Nanj ing Forestry University(Natural Science Edition),2003,27(3):11-14.DOI:10.3969/j.issn.1000-2006.2003.03.003.