中国野生植物资源2024,Vol.43Issue(10) :72-77.DOI:10.3969/j.issn.1006-9690.2024.10.009

延胡索叶片愈伤组织诱导及其2种活性成分含量测定

Callus Induction of Corydalis yanhusuo Leaf and Content Determination of Its Two Active Ingredients

梁宇莹 高靖靖 马阳阳 文莉 樊修和 于金高 颜永刚 李铂
中国野生植物资源2024,Vol.43Issue(10) :72-77.DOI:10.3969/j.issn.1006-9690.2024.10.009

延胡索叶片愈伤组织诱导及其2种活性成分含量测定

Callus Induction of Corydalis yanhusuo Leaf and Content Determination of Its Two Active Ingredients

梁宇莹 1高靖靖 2马阳阳 2文莉 2樊修和 1于金高 1颜永刚 2李铂1
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作者信息

  • 1. 陕西中医药大学 陕西中药资源产业化省部共建协同创新中心/秦药特色资源研究与开发国家重点实验室(培育),陕西 咸阳 712083
  • 2. 陕西中医药大学 药学院,陕西 咸阳 712046
  • 折叠

摘要

目的:以叶片为外植体诱导延胡索(Corydalis yanhusuo)形成愈伤组织,测定愈伤组织生长指标,检测生物碱类成分含量,为研究延胡索生物碱代谢调控机制和培育优良品系提供依据.方法:运用2,4-二氯苯氧乙酸(2,4-D)与6-苄基氨基嘌呤(6-BA)诱导形成愈伤组织,采用重量法测定生物量,组织化学染色法定位生物碱,高效液相色谱法测定延胡索乙素和脱氢紫堇碱含量.结果:延胡索叶片经MS+2 mg/L 2,4-D+1 mg/L 6-BA处理,愈伤组织诱导率可达75%;外观呈黄白色,表面呈颗粒状,生长迅速;继代培养的愈伤组织鲜重(2.7153±0.4398)g,增殖倍数4.43;干重(0.1552±0.0207)g,含水量94.16%.愈伤组织薄壁细胞内含能被碘化铋钾染色的橘黄色斑块,提示生物碱的存在;愈伤组织中延胡索乙素含量达(0.8600±0.3502)mg/g DW(干重),脱氢紫堇碱含量为(0.1744±0.0372)mg/g DW.结论:利用叶片作为外植体获得的延胡索愈伤组织诱导率高、生长迅速,是快速、稳定获取延胡索乙素等生物碱类活性成分的适宜材料.

Abstract

Objective:The leaf of Corydalis yanhusuo was used as explants to induce callus,the growth parameters and contents of alkaloids in the callus were measured,to provide a basis for the study of alkaloids metabolic regulation and superior varieties breeding of C.yanhusuo.Methods:The combination of 2,4-dichlorophenoxyacetic acid(2,4-D)and 6-benzyladenine(6-BA)were used for callus induction.The callus biomass was measured by weight method.Localization of alkaloids was performed by histochemical staining method.The contents of tetrahydropalmatine and dehydrocoryda-line were determined by high-performance liquid chromatography method.Results:The callus induc-tion rate of C.yanhusuo reached 75%by Murashige and Skoog medium with 2 mg/L 2,4-D and 1 mg/L 6-BA treatment.The callus was yellowish-white with a granular surface and grew rapidly.The fresh weight of subcultured callus was(2.7153±0.4398)g,with the multiplication coefficient of 4.43.The dry weight was(0.1552±0.0207)g,and the water content reached 94.16%.Callus parenchyma cells contained orange patches stained with Dragendorff's reagent,indicating the presence of alkaloids.The content of tetrahydropalmatine in the callus reached(0.8600±0.3502)mg/g DW(dry weight),meanwhile,dehydrocorydaline content reached(0.1744±0.0372)mg/g DW.Conclusion:The callus of C.yanhusuo induced by using leaves as explants showed a high induction rate and rapid growth,which was a suitable material for deriving alkaloids active ingredients such as tetrahydropalmatine quickly and stably.

关键词

延胡索/愈伤组织/生长指标/生物碱/延胡索乙素

Key words

Corydalis yanhusuo/Callus/Growth parameters/Alkaloids/Tetrahydropalmatine

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出版年

2024
中国野生植物资源
中华全国供销合作总社南京野生植物综合利用研究院

中国野生植物资源

影响因子:0.667
ISSN:1006-9690
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