摘要
【目的】探究温度胁迫对白桦丛生苗三萜、黄酮及抗逆酶积累的影响,为提高白桦次生代谢产物积累以及工厂化生产奠定基础。【方法】利用不同浓度(0.5、1.0、2.0和5.0 mg/L)的6-苄氨基腺嘌呤(6-BA),诱导产生白桦丛生苗及建立悬浮培养体系,同时对丛生苗进行温度胁迫诱导次生产物合成。【结果】附加2.0 mg/L 6-BA的NT固体培养基可诱导白桦愈伤组织获得白桦丛生苗。2种温度(45和4℃)胁迫下,白桦丛生苗中总三萜含量均比对照(25℃)有所提高,其中45℃处理4 h,恢复培养6 h后,总三萜含量比对照提高5.03倍。4℃处理白桦丛生苗1 h,并在24 h时取样,齐墩果酸含量比对照提高了14.52倍,最高含量达2.33 mg/g。4℃处理白桦丛生苗4 h,在恢复培养48 h时,黄酮含量比对照提高38.37%。同时明确温度胁迫下,4℃胁迫4 h,恢复培养72 h,超氧化物歧化酶(SOD)活性最高,比对照提高69.77%。经4℃处理1 h的丛生苗在96 h时,过氧化氢酶(CAT)活性达到最高峰,是对照的1.81倍。4℃处理1 h的丛生苗在恢复培养6 h时抗坏血酸过氧化物酶(APX)活性比对照提高55.29%。当胁迫时间延长至4 h,在45℃处理下,黄酮含量与APX酶,SOD与齐墩果酸含量均呈显著负相关(P<0.05);经4℃处理,总三萜与APX(P<0.01)、CAT(P<0.05)分别呈极显著正相关和显著正相关;黄酮与SOD含量呈显著负相关(P<0.05);CAT与APX(P<0.01)、SOD(P<0.05)相关性达极显著和显著水平,3种抗逆酶协调发挥作用共同参与次生产物合成。【结论】2.0 mg/L的6-BA能成功诱导白桦愈伤组织产生丛生苗,并建立悬浮培养系。短时间高温或低温胁迫均可以刺激3种防御酶发生显著变化,3种酶相互协调共同参与并促进白桦丛生苗总三萜和黄酮物质的合成与积累。
[Objective]The effects of temperature stress on the accumulation of triterpenoids,flavonoids and defense enzymes in multiple shoots of birch(Betula platyphylla)were investigated so as to lay the foundation for increasing the accumulation and factory production of secondary metabolites in birch.[Method]Different concentrations(0.5,1.0,2.0 and 5.0 mg/L)of 6-benzylaminoadenine(6-BA)were used to induce multiple shoots and establish suspension culture system of birch.Furthermore,multiple shoots were treated by temperature stress so that the synthesis of secondary metabolites in birch was induced.[Result]The callus of birch could be induced to obtain the multiple shoots by NT medium supplemented with 2.0 mg/L 6-BA.The content of total triterpenoids in birch multiple shoots under temperature stress(45 and 4℃)was higher than that of the control(25℃),among them the total triterpenoids content increased by 5.03 times after 4 h and then restoration culture for 6 h.Also the content of oleanolic acid in birch multiple shoots treated by 4℃for 1 h and sampled 24 h later was increased by 14.52 times,and even for 2.33 mg/g.And the content of flavonoids was enhanced by 38.37%under 4℃for 4 h and then 25℃for 48 h.Meanwhile,the superoxide dismutase(SOD)activity was highest under 4℃for 4 h and normal culture for 72 h,which was 69.77%higher than that of control,and the catalase(CAT)activity of birch multiple shoots treated by 4℃for 1 h and normal culture for 96 h reached the peak,which was increased to 1.81 times.Then the ascorbate peroxidase(APX)activity of multiple shoots treated at recovery culture for 6 h under 4℃for 1 h was 55.29%higher than that of control.While treated by 45℃for 4 h,the content of flavonoids and oleanolic acid was significantly negatively correlated(P<0.05)with APX enzyme and SOD activity,respectively.Furthermore,the content of total triterpenes was extremely significantly positively correlated or significantly positively correlated with APX activity(P<0.01)and CAT activity(P<0.05),and the cont
作者
李可鑫
赵微
徐林琳
李影
张宝莲
詹亚光
尹静
Li Kexin;Zhao Wei;Xu Linlin;Li Ying;Zhang Baolian;Zhan Yaguang;Yin Jing(Key Laboratory of Saline-Alkali Vegetation Ecology Restoration of Ministry of Education,College of Life Sciences,Northeast Forestry University,Harbin 150040,Heilongjiang,China;Landscaping Construction and Maintenance Center of Longjiang County,Qiqihar 161000,Heilongjiang,China)
出处
《北京林业大学学报》
CAS
CSCD
北大核心
2021年第7期31-39,共9页
Journal of Beijing Forestry University
基金
国家自然科学基金项目(31570589)
中央高校基本科研业务费专项(2572020DY22)
东北林业大学大学生创新项目(201910225082)。
关键词
白桦
丛生苗
悬浮培养
6-BA
次生产物
防御酶
Betula platyphylla
multiple shoots
suspension culture
6-BA
secondary product
defensive enzyme
