沙棘胚状体诱导分化技术研究
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摘要
[目的]探究不同基因型、基本培养基、激素种类及激素浓度对沙棘胚性愈伤组织诱导和胚状体分化的影响,为体胚发生体系的建立提供科学依据。[方法]将中国沙棘和蒙古沙棘种子经5d暗培养后在(14L∶10D)h·d-1的光周期下培养15d,获得的无菌试验材料,并将种胚的各个结构为外植体进行胚性愈伤组织诱导,用子叶和下胚轴以L9(34)正交设计进行胚性愈伤组织培养条件优化以及胚状体分化诱导。[结果]试验范围内,中国沙棘种子培养的最佳培养基是1/2 MS;由中国沙棘和蒙古沙棘种子培养20d所得的无菌子叶连带0.2cm下胚轴是胚性愈伤组织和胚状体诱导分化的良好材料。[结论]最适合中国沙棘和蒙古沙棘胚性愈伤组织诱导的培养基为:1/3 MS+6-BA0.5mg·L-1+IBA 1.0mg·L-1,诱导率可达75.9%;诱导胚状体分化的最佳培养基为:1/3 MS+IBA 0.05mg·L-1+KT 0.1mg·L-1+6-BA 1.0mg·L-1,分化率可达66%。
[Objective]The purpose of the present study was to determine the effect of various genetic types,basic culture media,and the type and concentration of certain hormone on the differentiation and developmental rate of embryogenic callus of Hippophaerhamnoides,and to provide scientific support for the establishment of somatic embryogenesis.[Methods]The seeds of H.rhamnoidessubsp.sinensis and H.rhamnoides subsp.mongolica were growing under completely dark condition for 5 days,and a photoperiod of 14∶10 hL∶D for 15 d.Sterile cotyledon,hypocotyl,and taproot were collected and used for inducing embryogenic callus and embryoid.Conditions were optimized with L9(34)Orthogonal Design Test.[Results]Under study conditions,the best cultural medium for the seeds of H.rhamnoides subsp.Sinensis was 1/2 MS,and the 20-day old sterile cotyledons with 0.2 cm-hypocotyl of both two species of H.rhamnoides were determined as the best material for embryogenic callus induction and differentiation.[Conclusion]The optimal medium for the embryogenic callus induction was 1/3 MS+6-BA 0.5 mg·L-1+IBA 1.0 mg·L-1,and the highest induction rate was reached up to 75.9%.The optimal medium for the somatic embryogenesis induction was 1/3 MS+IBA 0.05 mg·L-1+KT 0.1 mg·L-1+6-BA 1.0 mg·L-1 with the rate at 66%.
[Objective]The purpose of the present study was to determine the effect of various genetic types,basic culture media,and the type and concentration of certain hormone on the differentiation and developmental rate of embryogenic callus of Hippophaerhamnoides,and to provide scientific support for the establishment of somatic embryogenesis.[Methods]The seeds of H.rhamnoidessubsp.sinensis and H.rhamnoides subsp.mongolica were growing under completely dark condition for 5 days,and a photoperiod of 14∶10 hL∶D for 15 d.Sterile cotyledon,hypocotyl,and taproot were collected and used for inducing embryogenic callus and embryoid.Conditions were optimized with L9(34)Orthogonal Design Test.[Results]Under study conditions,the best cultural medium for the seeds of H.rhamnoides subsp.Sinensis was 1/2 MS,and the 20-day old sterile cotyledons with 0.2 cm-hypocotyl of both two species of H.rhamnoides were determined as the best material for embryogenic callus induction and differentiation.[Conclusion]The optimal medium for the embryogenic callus induction was 1/3 MS+6-BA 0.5 mg·L-1+IBA 1.0 mg·L-1,and the highest induction rate was reached up to 75.9%.The optimal medium for the somatic embryogenesis induction was 1/3 MS+IBA 0.05 mg·L-1+KT 0.1 mg·L-1+6-BA 1.0 mg·L-1 with the rate at 66%.
引文
[1]李永霞,王照程,杨小林.江孜沙棘种子萌发特性研究[J].种子,2018,37(3):86-88.
[2]刘丽,王莉,阮成江.沙棘种子萌发的基本培养基的优化[J].大连民族大学学报,2016,18(5):469-472.
[3]徐虹,梁宗锁.沙棘组织培养技术的研究[J].西北植物学报,2001,21(2):267-272.
[4]李燕.秦艽发根系体细胞胚的诱导和植株再生[D].兰州:兰州大学,2008.
[5]孙政,陈发菊,高晗,等.楸树胚性愈伤组织与非胚性愈伤组织的生理生化差异[J].分子植物育种,2017,15(11):4642-4646.
[6]王雪丽,李萍萍,吴坤阳,等.日本五针松针叶胚性愈伤诱导及愈伤组织细胞学特性观察[J].植物生理学报,2017,53(3):422-428.
[7]刘艳军,张超,杨静慧,等.塔杨松散型均质胚性愈伤组织培养体系[J].植物研究,2016,36(1):123-128.
[8]肖望,涂红艳,张爱玲.白姜花胚性愈伤组织的诱导与植株再生体系的建立[J].园艺学报,2016,43(8):1605-1612.
[9]陈琴,唐红亮,梁机,等.利用广西杉木良种未成熟胚诱导胚性愈伤组织发生[J].基因组学与应用生物学,2017,36(5):2076-2081.
[10]Sherif N A,Benjamin J H F,Kumar T S,et al.Somatic embryogenesis,acclimatization and genetic homogeneity assessment of regenerated plantlets of Anoectochilus elatus Lindl.,an endangered terrestrial jewel orchid[J].Plant Cell Tissue and Organ Culture,2018,132(2):303-316.
[11]Mamedes-Rodrigues T C,Batista,D S,Vieira N M.Regenerative potential,metabolic profile,and genetic stability of Brachypodium distachyon embryogenic calli as affected by successive subcultures[J].Protoplasma,2018,255(2):655-667.
[12]Lin H S,Toorn C V D,Raemakers K J J M,et al.Development of a plant regeneration system based on friable embryogenic callus in the ornamental Alstroemeria[J].Plant Cell Reports,2000,19(5):529-534.
[13]高洁,王元忠,黄衡宇.绿花百合胚性愈伤组织诱导与植株再生研究[J].植物研究,2016,36(1):52-57.
[14]刘志红,李周岐.接骨木茎段组织培养技术研究[J].西北林学院学报,2008,23(6):101-103.
[15]赵晓敏,沈海龙,杨玲,等.兴安落叶松胚性愈伤组织诱导影响因子的研究[J].植物研究,2007,27(5):538-543.
[16]周思宇,王永清.枇杷叶片胚性愈伤组织诱导与内源激素含量的关系[J].植物科学学报,2017,35(1):99-106.
[17]Xiao Y Q,Chen Y L,Ding Y P,et al.Effects of GhWUS from upland cotton(Gossypium hirsutwn L.)on somatic embryogenesis and shoot regeneration[J].Plant Science,2018,270:157-165.
[18]Ebrahimi M,Mokhtari A,Amirian R.A highly efficient method for somatic embryogenesis of Kelussia odorotissima Mozaff.,an endangered medicinal plant[J].Plant Cell Tissue and Organ Culture,2018,132(01):99-110.
[19]Borji M,Bouamama-Gzara B,Chibani Farhat,et al.Micromorphology,structural and ultrastructural changes during somatic embryogenesis of a Tunisian oat variety(Avena sativa L.var‘Meliane’)[J].Plant Cell Tissue and Organ Culture,2018,132(2):329-342.
[20]王征,伍江波,刘雪梅,等.杜仲愈伤组织诱导和植株再生体系的建立[J].西北农林科技大学学报:自然科学版,2015,43(7):57-65.
[21]张健欣,唐红.不同外源激素对紫斑牡丹愈伤组织诱导的影响[J].甘肃农业大学学报,2015,50(6):74-80.
[22]柳玉晶,龚束芳,樊金萍,等.百合愈伤组织的诱导及植株再生[J].东北农业大学学报,2007,38(3):352-355.
[2]刘丽,王莉,阮成江.沙棘种子萌发的基本培养基的优化[J].大连民族大学学报,2016,18(5):469-472.
[3]徐虹,梁宗锁.沙棘组织培养技术的研究[J].西北植物学报,2001,21(2):267-272.
[4]李燕.秦艽发根系体细胞胚的诱导和植株再生[D].兰州:兰州大学,2008.
[5]孙政,陈发菊,高晗,等.楸树胚性愈伤组织与非胚性愈伤组织的生理生化差异[J].分子植物育种,2017,15(11):4642-4646.
[6]王雪丽,李萍萍,吴坤阳,等.日本五针松针叶胚性愈伤诱导及愈伤组织细胞学特性观察[J].植物生理学报,2017,53(3):422-428.
[7]刘艳军,张超,杨静慧,等.塔杨松散型均质胚性愈伤组织培养体系[J].植物研究,2016,36(1):123-128.
[8]肖望,涂红艳,张爱玲.白姜花胚性愈伤组织的诱导与植株再生体系的建立[J].园艺学报,2016,43(8):1605-1612.
[9]陈琴,唐红亮,梁机,等.利用广西杉木良种未成熟胚诱导胚性愈伤组织发生[J].基因组学与应用生物学,2017,36(5):2076-2081.
[10]Sherif N A,Benjamin J H F,Kumar T S,et al.Somatic embryogenesis,acclimatization and genetic homogeneity assessment of regenerated plantlets of Anoectochilus elatus Lindl.,an endangered terrestrial jewel orchid[J].Plant Cell Tissue and Organ Culture,2018,132(2):303-316.
[11]Mamedes-Rodrigues T C,Batista,D S,Vieira N M.Regenerative potential,metabolic profile,and genetic stability of Brachypodium distachyon embryogenic calli as affected by successive subcultures[J].Protoplasma,2018,255(2):655-667.
[12]Lin H S,Toorn C V D,Raemakers K J J M,et al.Development of a plant regeneration system based on friable embryogenic callus in the ornamental Alstroemeria[J].Plant Cell Reports,2000,19(5):529-534.
[13]高洁,王元忠,黄衡宇.绿花百合胚性愈伤组织诱导与植株再生研究[J].植物研究,2016,36(1):52-57.
[14]刘志红,李周岐.接骨木茎段组织培养技术研究[J].西北林学院学报,2008,23(6):101-103.
[15]赵晓敏,沈海龙,杨玲,等.兴安落叶松胚性愈伤组织诱导影响因子的研究[J].植物研究,2007,27(5):538-543.
[16]周思宇,王永清.枇杷叶片胚性愈伤组织诱导与内源激素含量的关系[J].植物科学学报,2017,35(1):99-106.
[17]Xiao Y Q,Chen Y L,Ding Y P,et al.Effects of GhWUS from upland cotton(Gossypium hirsutwn L.)on somatic embryogenesis and shoot regeneration[J].Plant Science,2018,270:157-165.
[18]Ebrahimi M,Mokhtari A,Amirian R.A highly efficient method for somatic embryogenesis of Kelussia odorotissima Mozaff.,an endangered medicinal plant[J].Plant Cell Tissue and Organ Culture,2018,132(01):99-110.
[19]Borji M,Bouamama-Gzara B,Chibani Farhat,et al.Micromorphology,structural and ultrastructural changes during somatic embryogenesis of a Tunisian oat variety(Avena sativa L.var‘Meliane’)[J].Plant Cell Tissue and Organ Culture,2018,132(2):329-342.
[20]王征,伍江波,刘雪梅,等.杜仲愈伤组织诱导和植株再生体系的建立[J].西北农林科技大学学报:自然科学版,2015,43(7):57-65.
[21]张健欣,唐红.不同外源激素对紫斑牡丹愈伤组织诱导的影响[J].甘肃农业大学学报,2015,50(6):74-80.
[22]柳玉晶,龚束芳,樊金萍,等.百合愈伤组织的诱导及植株再生[J].东北农业大学学报,2007,38(3):352-355.