通过多种培养基优化长白落叶松体细胞胚再生  

Optimization of Somatic Embryo Regeneration of Larix olgensis Through Multiple Media

作　　者：刘威锋 侯佳音 蒋湘宁[1] Liu Weifeng;Hou Jiayin;Jiang Xiangning(College of Biology,Beijing ForestryUniversity,Beijing,100083)

机构地区：[1]北京林业大学生物学院,北京100083

出　　处：《分子植物育种》2021年第15期5083-5091,共9页Molecular Plant Breeding

基　　金：国家科技专项(2018ZX08020003-001-002)资助

摘　　要：为建立稳定高效的落叶松体细胞胚再生体系,本研究以长白落叶松未成熟合子胚作为外植体材料,选择生长良好的胚性愈伤组织,依次通过液体培养基A,半固体培养基B,固体培养基C三种成熟培养基诱导体细胞胚形成。其中,愈伤诱导培养基为S培养基+2,4-D 1.0 mg/L+6-BA0.5 mg/L+KT 0.3 mg/L,蔗糖20 g/L,琼脂3 g/L;继代培养基为S培养基+2,4-D0.15 mg/L+6-BA 0.05 mg/L+KT 0.05 mg/L,蔗糖30 g/L,琼脂6 g/L;成熟培养基A为S培养基+PEG-4000100 g/L+ABA 15 mg/L+AgNO310 mg/L,蔗糖45 g/L的液体培养基;成熟培养基B和C分别为添加了2 g/L和10 g/L琼脂的成熟培养基A。该方法体胚成熟时间为32 d左右,同步率为78.0%,正常形态体胚比例为83.0%。正常形态体胚形成效率可达223.0个/g胚性愈伤,体细胞胚萌发率可达47.2%。相较于单一成熟培养基,缩短成熟时间,使体胚生成更同步,并提高了体胚产量和质量。为落叶松体胚再生的商业化提供了途径,为大规模化育种繁殖、生理生化研究和转基因改良带来极大的方便,具有较大实际应用价值。To establish a stable and e fficient Larix olgensis somatic embryo regeneration system.In this study,immature zygotic embryos of Larix olgensis were used as explant materials for embryogenic callus induction,and embryogenic callus with good growth were selected to induce somatic embryo formation by liquid medium A,semi-solid medium B,and solid medium C.The callus induction medium is S medium+2,4-D 1.0 mg/L+6-BA 0.5 mg/L+KT 0.3 mg/L,sucrose 20 g/L,agar 3 g/L;the subculture medium is S medium+2,4-D 0.15 mg/L+6-BA 0.05 mg/L+KT 0.05 mg/L,sucrose 30 g/L,agar 6 g/L.Mature medium A is S medium+PEG-4000100 g/L+ABA 15 mg/L+AgNO310 mg/L,sucrose 45 g/L liquid medium.Mature medium B and C are mature medium A supplemented with 2 g/L and 10 g/L agar,respectively.The maturation time of this method is about 32 days,the syn-chronization rate is 78.0%,and the proportion of somatic embryos in normal morphology is 83.0%.Somatic embryo formation efficiency in normal morphology can reach 223.0 embryos/g embryogenic callus,somatic embryo germination rate can reach 47.2%.Compared with a single maturation medium,the maturation time is shortened,somatic embryo generation is more synchronized,and high somatic embryo yield and quality are improved.It pro-vides a way for the commercialization of somatic embryo regeneration of Larix olgensis,brings great convenience to large-scale breeding and reproduction,physiological and biochemical research,and genetic modification,and has great practical application value.

关 键 词：长白落叶松 胚性愈伤 体细胞胚发生 

分 类 号：S791.22[农业科学—林木遗传育种]