胡桃楸组培过程中茎段褐化机理初步分析  

Preliminary Analysis on Browning Mechanism of Juglans mandshurica Stem in Tissue Culture

作　　者：李佳[1] 王斯彤 周强 杨雨春[4] 祁永会 王钦美[1] 张丽杰[1] Li Jia;Wang Sitong;Zhou Qiang;Yang Yuchun;Qi Yonghui;Wang Qinmei;Zhang Lijie(Key Laboratory of forest tree genetics,breeding and Cultivation of Liaoning Province/College of Forestry,Shenyang Agricultural University,Shenyang,110866;Liaoning Institute of Sand Control and Utilization,Fuxin,123000;Liaoning Forestry Development Service Center,Shenyang,110036;Jilin Provincial Academy of Forestry Science,Changchun,130033;Forestry Research Institute of Heilongjiang Province,Harbin,150040)

机构地区：[1]沈阳农业大学林学院,辽宁省林木遗传育种与培育重点实验室,沈阳110866 [2]辽宁省沙地治理和利用研究所,阜新123000 [3]辽宁省林业发展服务中心,沈阳110036 [4]吉林省林业科学研究院,长春130033 [5]黑龙江省林业科学研究所,哈尔滨150040

出　　处：《分子植物育种》2021年第24期8239-8244,共6页Molecular Plant Breeding

基　　金：国家重点研发计划项目(2017YFD0600605);辽宁省自然科学基金项目(20170540801)。

摘　　要：为探讨胡桃楸(Juglans mandshurica)茎段在离体培养过程中褐化发生的机理。在培养基中添加不同浓度和种类的抗褐化剂,对不同培养时间的胡桃楸茎段进行生化指标测定和徒手切片染色观察。在初代培养时外植体采用70%酒精30 s,0.1%Hg Cl;消毒10 min,灭菌效果最好。在初代培养基中添加不同浓度和种类的抗褐化剂可以有效地防止外植体培养过程中发生褐化,并筛选出同时添加硫代硫酸钠和Vc两种试剂的培养基抗褐化能力最强。通过测定不同培养阶段茎段体内酶活性,结果发现随着培养时间的延长,褐化茎段和未褐化茎段体内PPO、POD、SOD、MDA活性均呈现下降趋势,且经过2周离体培养后茎段内各种酶活性开始下降,并逐渐失活而最终导致外植体褐变死亡。这一变化过程与徒手切片染色观察结果相一致。说明胡桃楸茎段培养过程中褐化的发生与多酚类物质和酶活性密切相关。本研究初步阐明了胡桃楸茎段在离体培养过程中发生褐化的原因,为进一步研究胡桃楸组织培养获得再生植株提供一定参考。To study the mechanism of browning of Juglans mandshurica stems in vitro culture.Different concentrations and kinds of anti-browning agents were added to the culture medium,and biochemical indexes of Juglans mandshurica stem segments at different culture times were determined and observed by unarmed section staining.In the primary culture,the explants were sterilized with 70%alcohol for 30 s and 0.1%Hg Cl2 for 10 min,with the best effect.Adding different concentrations and kinds of anti browning agents to the primary culture medium can effectively prevent browning in the process of explant culture,and screening out that adding sodium thiosulfate and Vc at the same time to the culture medium has the strongest anti browning ability.The results showed that the activities of PPO,POD,SOD and MDA in browning and non browning stem segments decreased with the prolongation of culture time,and after 2 weeks in vitro culture,the activities of various enzymes in stem segments began to decline,and gradually inactivated,leading to the browning death of explants.With the result of cell structure.The browning of Juglans mandshurica stem is closely related to polyphenols and enzyme activities.This study preliminarily elucidated the reason of browning of stem explants of Juglans mandshurica in vitro culture,and provided some reference for further study on tissue of Juglans mandshurica to obtain regenerated plants.

关 键 词：胡桃楸 幼嫩茎段 离体培养 褐变机理 

分 类 号：S792.132[农业科学—林木遗传育种]