植物研究2025，Vol.45Issue(4)：533-545,13.DOI:10.7525/j.issn.1673-5102.2025.04.006
二倍体青钱柳愈伤组织诱导及次生代谢物积累
Callus Induction and Secondary Metabolites Accumulation of Diploid Cyclocarya paliurus
摘要
Abstract
This study aimed to explore the effects of hormone combination on callus induction and proliferation of diploid Cyclocarya paliurus and the accumulation of main secondary metabolites during subculture.The effects of different hormone combinations on accumulation of secondary metabolites during callus induction and subproliferation were investigated using diploid C.paliurus leaves as explants by orthogonal experiment design.An optimal treatment for the maximum yields of total triterpenoids,total polyphenols,total flavonoid and crude polysaccharides were selected by entropy weight method.The highest callus induction rate was 100%in treatment 2(0.5 mg•L-1 6-BA+1 mg•L-1 2,4-D+0.2 mg•L-1 NAA),followed by treatment 9(2.5 mg•L-1 6-BA+1.5 mg•L-1 2,4-D+0.2 mg•L-1 NAA)and treatment 5(1.5 mg•L-1 6-BA+1 mg•L-1 2,4-D+0.4 mg•L-1 NAA),with an induction rate of 96.67%and 93.64%,respectively.The order of factors affecting callus induction rate was 2,4-D,NAA,6-BA.An optimal concentration of 2,4-D and NAA was 1.0 mg•L-1 and 0.2 mg•L-1,with no significant effect of 6-BA.The colors of induced callus were light green and yellowish green;the morphology of the callus was characterized by loose texture.There were differences in the callus growth and accumulation of secondary metabolites among treatments when different combinations of hormones were used for subculture;the growth characteristics of calli in the same treatment changed greatly during continuous subculture.The first ranked formula was treatment 4(0.2 mg•L-1 KT+0.5 mg•L-1 6-BA+0.75 mg•L-1 NAA+0.4 mg•L-1 2,4-D)with a score of 0.44,followed by treatment 2(0.75 mg•L-1 6-BA+0.75 mg•L-1 NAA+0.2 mg•L-1 2,4-D)and treatment 6(0.2 mg•L-1 KT+1.0 mg•L-1 6-BA+0.5 mg•L-1 NAA+0.2 mg•L-1 2,4-D)with a score of 0.31 and 0.26,respectively.The yield of secondary metabolites in treatment 4 generally reached a high level from the 8th generation,with total triterpenoid yield of 28.91 mg•bottle-1,total polyphenol yield of 6.89 mg•bottle-1,total flavonoid yield of 1.77 mg•bottle-1,and crude polysaccharide yield of 57.45 mg•bottle-1.In summary,the optimal formula for callus induction of diploid C.paliurus was MS+0.5 mg•L-1 6-BA+1.0 mg•L-1 2,4-D+0.2 mg•L-1 NAA.The subculture proliferation formula was MS+0.2 mg•L-1 KT+0.5 mg•L-1 6-BA+0.75 mg•L-1 NAA+0.4 mg•L-1 2,4-D.This study provided a reference for production of secondary metabolites by callus culture and further cell suspension culture.关键词
青钱柳/愈伤组织诱导/植物激素/继代增殖/次生代谢物Key words
Cyclocarya paliurus/callus induction/plant hormone/subculture proliferation/secondary metabolites分类
生物科学引用本文复制引用
李卓穗,高一琳,刘寒,尚旭岚..二倍体青钱柳愈伤组织诱导及次生代谢物积累[J].植物研究,2025,45(4):533-545,13.基金项目
国家自然科学基金项目(31971642). （31971642）
