抗性湿地松胚性愈伤组织的维持与增殖  

Maintenance and Proliferation of Resistant Pinus elliottii Embryonic Callus

作　　者：程方[1,2] 叶建仁 Cheng Fang;Ye Jianren(Co-Innovation Center for Sustainable Forestry in Southern China,Nanjing Forest University,Nanjing,Jiangsu 210037,China;Guangdong Xiangtoushan National Nature Reserve Administration,Huizhou,Guangdong 516001,China)

机构地区：[1]南京林业大学南方现代林业协同创新中心,江苏南京210037 [2]广东象头山国家级自然保护区管理局,广东惠州516001

出　　处：《绿色科技》2023年第14期1-7,27,共8页Journal of Green Science and Technology

基　　金：“十四五”国家重点研发项目(编号:2021YFD140090)。

摘　　要：将抗松针褐斑病湿地松愈伤组织转接至不同培养基及多种添加物中,开展了不同愈伤组织类型、不同浓度肌醇与麦芽糖、不同2,4-二氯苯氧乙酸(2,4-D)、6-苄基氨基嘌呤(6-BA)、激动素(KT)浓度组合、添加维生素(VC)、添加硝酸银(AgNO3)、不同基因型胚性愈伤组织、多次继代后胚性愈伤组织与培养基的适应性等研究。结果表明:湿地松不同愈伤组织类型在增殖过程中成活率及增殖状况都存在很大差异;当麦芽糖浓度为15 g/L,肌醇浓度为1000 mg/L时最有利于胚性胚柄细胞团(ESM)的增殖;高浓度的2,4-D、6-BA不利于湿地松胚性愈伤组织的增殖,最佳增殖培养基及激素组合DCR+1.0 mg/LNAA+0.32 mg/L6-BA+0.31 mg/LKT、DCR+1.0 mg/LNAA(萘乙酸)+1.55 mg/L6-BA+0.064 mg/LKT;添加维生素(VC)浓度为10 mg/L时,对湿地松愈伤的褐化有明显抑制作用,但浓度过高则会增加愈伤的褐化率;添加3.4 mg/L的AgNO3促进了湿地松胚性愈伤组织的增殖;湿地松不同家系不同基因型间增殖能力差异显著,20#2和10#1增殖很快明显优于10#4、10#5和20#3;湿地松胚性胚柄细胞团在同样的培养基上多次继代后,愈伤组织增殖变慢,中心褐化较多,有些还会出现水渍状,可能导致其胚性丧失。湿地松不同愈伤组织类型在增殖过程中,成活率及增殖状况都存在很大的差异。Ⅲ型、Ⅳ型愈伤可能同时含有胚性愈伤和非胚性愈伤,在培养基条件适合的情况下胚性愈伤转化成胚性胚柄细胞团,但是转化频率极低。而Ⅴ型、Ⅵ型愈伤在后期培养过程中基本褐化死亡。In this study,Pinus elliottii embryonic callus embryonic of resistant Lecanosticta acicola were transferred to different culture media and various additives to explore different callus types,different concentrations of inositol and maltose,different 2,4-dichlorophenoxyacetic acid(2,4-D),6-Benylaminopurine(6-BA),kinetin(KT)concentration combination,added vitamin(VC),added silver nitrate(AgNO 3),different genotypes of embryogenic callus,multiple generations of embryogenic research on the adaptability of callus and culture medium.The results showed that there were great differences in the survival rate and proliferation status of different callus types of Pinus elliottii during the proliferation process;when the maltose concentration was 15g/L and the inositol concentration was 1000mg/L,it was most beneficial to the Embryonal-suspensor mass(ESM)proliferation;high concentration of 2,4-D,6-BA was not conducive to the proliferation of Pinus elliottii embryogenic callus;the best proliferation medium and hormone combination is DCR+1.0mg/LNAA+0.32mg/L6-BA+0.31mg/LKT,DCR+1.0 mg/LNAA(a-naphthalene acetic acid)+1.55mg/L6-BA+0.064mg/LKT;when the concentration of vitamin(VC)was 10 mg/L,the browning of Pinus elliottii callus had an obvious inhibitory effect,but if the concentration was too high,it would increase the browning rate of the callus;the addition of 3.4 mg/L AgNO 3 promoted the proliferation of Pinus elliottii embryogenic callus;the different genes of Pinus elliottii were different.There were significant differences in proliferation ability between types;20#2 and 10#1 proliferate quickly and are significantly better than 10#4,10#5 and 20#3;Pinus elliottii embryonic stalk cell clusters were subcultured on the same medium for many times.Later,the callus proliferation slowed down.The center browns more,and some would appear water-stained,which may lead to the loss of its embryonicity.TypeⅢand typeⅣcallus may contain both embryogenic callus and non-embryogenic callus.Embryogenic callus transformed into embryogenic emb

关 键 词：抗性湿地松 体细胞胚胎发生 胚性愈伤组织 维持与增殖培养基 

分 类 号：S791[农业科学—林木遗传育种]