长白落叶松胚性愈伤组织诱导及体细胞胚胎发生  

Embryogenic Callus Induction and Somatic Embryogenesis from Immature Zygotic Embryos of Larix olgensis

作　　者：宋跃[1] 甄成[1] 张含国[1] 李淑娟[1] 

机构地区：[1]林木遗传育种国家重点实验室(东北林业大学),哈尔滨150040

出　　处：《林业科学》2016年第10期45-54,共10页Scientia Silvae Sinicae

基　　金：国家"863"计划项目(2013AA102704-04-03);东北林业大学林木遗传育种国家重点实验室创新项目(2015B03)

摘　　要：【目的】以长白落叶松的未成熟合子胚为外植体,进行胚性愈伤组织的诱导、增殖培养及体细胞胚胎发生的相关研究,揭示影响长白落叶松胚性愈伤组织诱导的关键因素,并探讨继代培养过程中添加不同种类、浓度的生长调节剂对胚性愈伤组织增殖及体胚发生的影响。【方法】采集长白落叶松3个家系优良单株的种子,诱导胚性愈伤组织,比较外植体采集时间、家系、2,4-D浓度及基本培养基对胚性愈伤组织诱导的影响。诱导出的胚性愈伤组织继代于含不同种类、浓度生长调节剂的增殖培养基,通过体胚发生途径获得体细胞胚。选取发育正常的体胚进行萌发,待体胚生根后移栽。【结果】不同时间采集的未成熟合子胚,其胚性愈伤组织诱导率存在较大差别,授粉后63天的合子胚诱导率为5.61%,授粉后70天诱导率为22.35%,而授粉后80天未诱导出胚性愈伤组织;‘长77-22’、‘长77-37’和‘长73-50’3个家系胚性愈伤组织的平均诱导率分别为6.69%,11.17%和3.11%;2,4-D浓度对长白落叶松胚性愈伤组织诱导具有一定影响,在一定范围内胚性愈伤组织的诱导率会随2,4-D浓度升高而升高,当其浓度为1.5 mg·L^(-1)时胚性愈伤组织的诱导率最高,达到11.11%,而当2,4-D浓度超过1.5 mg·L^(-1)时,胚性愈伤组织的诱导率则开始下降;BM,MS,S培养基均能诱导出胚性愈伤组织,其中,BM培养基的诱导率最高,S培养基的诱导率次之,MS培养基的诱导率最低。胚性愈伤组织在含2,4-D 0.3 mg·L^(-1)、6-BA0.1 mg·L^(-1)及KT 0.1 mg·L^(-1)的BM培养基上增殖15天可获得相对较多的胚性愈伤组织,增殖率为345.93%;在含2,4-D 1.5 mg·L^(-1)、6-BA 0.5 mg·L^(-1)及KT 0.5 mg·L^(-1)的BM培养基上培养14天,再经诱导可获得较多的体细胞胚胎,每克愈伤组织平均诱导179.87个体胚,并且这些体胚的萌发率及植株再生率相对较高,分别为75.00%,66.67%;再生植株移栽成活�【Objective 】 Immature zygotic embryos of Larix olgensis were used as explants to study induction,proliferation and somatic embryogenesis of embryogenic callus in order to reveal the key factors affecting Larix olgensis embryogenic callus induction,and at the same time,to explore the effects of growth regulators of different types and concentrations on proliferation and somatic embryogenesis of embryogenic callus,in subculture process. 【Method 】Embryogenic callus was induced using immature seeds from superior individuals of three families of Larix olgensis,and the effects of seed collection time,family,concentration of 2,4-D and basic medium on the induction of embryogenic calluswere studied. Subsequently,the embryogenic callus subculture on the proliferation medium containing different kinds and concentrations of growth regulators,and the somatic embryos were obtained by the process of somatic embryogenesis.Finally,morphologically normal somatic embryos were selected and germinated,and transplanted after rooting of the somatic embryos. 【Result】There was significant differences in induction rate of embryonic callus of immature embryos with different collection times. The induction rate was 5. 61% 63 days after pollination,the induction rate was 22. 35%70 days after pollination,and the induction rate was zero 80 days after pollination. In this experiment,the average induction rate of embryogenic callus from the families of ‘77-22＇,‘77-37 ＇and ‘73-50 ＇was 6. 69%,11. 17% and3. 11%,respectively. The concentration of 2,4-D had a certain effect on the induction of embryogenic callus,the induction rate was increased with the proliferated of 2,4-D concentration in a certain range. It was up to 11. 11% when the concentration of 2,4-D reached 1. 5 mg·L-（-1）. The induction rate began to decrease when the concentration of 2,4-D exceeded 1. 5 mg·L-（-1）. The medium of BM,MS and S were able to induce embryogenic callus,the induction rate in BM medium was the highest,followed by S medium,and the

关 键 词：长白落叶松 体胚诱导 胚性愈伤组织 体细胞胚胎 植株再生 

分 类 号：S718.46[农业科学—林学]