核桃叶片愈伤组织的诱导及植株再生研究
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摘要
以杂种核桃组培苗的叶片和叶柄为材料,设置5种激素组合进行叶片和叶柄愈伤组织的诱导,并设置4种激素组合进行愈伤组织再分化的诱导,以研究杂种核桃的组培再生技术。结果表明:叶柄较叶片更容易发生愈伤组织,并且褐化率较低;TDZ 4mg·L~(-1)+2,4-D 1.5mg·L~(-1)和6-BA 0.5mg·L~(-1)+NAA 0.5mg·L~(-1)+TDZ1.0mg·L~(-1)2种植物激素组合最适合愈伤组织的诱导。产生的愈伤组织呈白色和黄白色,为非胚性愈伤组织。在4种激素组合下愈伤组织器官,但均未诱导成功。部分愈伤组织有增殖,培养1个月后全部褐化。研究说明杂种核桃的愈伤组织较容易诱导发生,而器官的再分化较困难。
In order to study the tissue culture regeneration technology of hybrid walnut,in this study we used the leaves and petioles of hybrid walnut tissue culture seedlings as materials,and 5 hormone combinations were set to induce callus of leaves and petioles. In addition,4 hormone combinations were set to induce callus redifferentiation.The results showed that the petioles was more easy to induce callus than leaves,and the browning rate was lower. We found two hormone combinations which TDZ4 mg·L~(-1)+2,4-D1.5 mg·L~(-1) and 6-BA0.5 mg·L~(-1)+NAA0.5 mg·L~(-1)+TDZ1.0 mg·L~(-1) were the most suitable for callus induction. The callus induced was white and yellowish white,but it was non-embryonic callus. Organ induction from callus was all not successful under the combinations of 4 hormones.Some of the callus proliferated and all browned after cultured one month. It showed that callus of hybrid walnut was easy to induce,but the redifferentiation of organs was difficult. The results of this study laid a certain foundation for further study on walnut callus organ differentiation.
In order to study the tissue culture regeneration technology of hybrid walnut,in this study we used the leaves and petioles of hybrid walnut tissue culture seedlings as materials,and 5 hormone combinations were set to induce callus of leaves and petioles. In addition,4 hormone combinations were set to induce callus redifferentiation.The results showed that the petioles was more easy to induce callus than leaves,and the browning rate was lower. We found two hormone combinations which TDZ4 mg·L~(-1)+2,4-D1.5 mg·L~(-1) and 6-BA0.5 mg·L~(-1)+NAA0.5 mg·L~(-1)+TDZ1.0 mg·L~(-1) were the most suitable for callus induction. The callus induced was white and yellowish white,but it was non-embryonic callus. Organ induction from callus was all not successful under the combinations of 4 hormones.Some of the callus proliferated and all browned after cultured one month. It showed that callus of hybrid walnut was easy to induce,but the redifferentiation of organs was difficult. The results of this study laid a certain foundation for further study on walnut callus organ differentiation.
引文
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[6]韩婷婷,孙周平.矮丛蓝莓叶片的愈伤组织诱导及植株再生[J].西北植物学报,2010,30(3):0615-0620.
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[10]李好先.核桃遗传转化体系的建立及rolB基因的导入[D].北京:中国农业科学院,2013.
[11]张燕,范宏伟,张国强,等.汾阳核桃叶片愈伤组织的诱导、增殖及分化培养[J].中国农学通报,2006,22(8):91-91.
[12]Heile-Sudholt C,Huetteman C A,Preece J E,Van Sambeek J W,Gaffney G R,In vitro embryonic axis and seedling shoot tip culture of Juglans nigra L[J]. Plant Cell Tissue Organ Cult. 1986,(6):189–197.
[13]李好先,曹尚银,薛辉,等.‘绿波’核桃叶片愈伤组织的诱导研究[J].分子植物育种,2015,(5):1060-1066.
[2]张志华,裴东.核桃学[M].北京:中国农业出版社,2018.
[3]郗荣庭,张毅萍.中国核桃[M].北京:中国林业出版社,1992.
[4]刘艳丽.柚木无性系愈伤组织诱导及植株再生[D].北京:中国林业科学研究院,2014.
[5]陈宗礼,张向前,齐向英,等.枣树叶片愈伤组织培养与再生植株的研究[J].福建林业科技,2007,3(43):23-26,35.
[6]韩婷婷,孙周平.矮丛蓝莓叶片的愈伤组织诱导及植株再生[J].西北植物学报,2010,30(3):0615-0620.
[7]Chalupa V,Clonal propagation of broad-leaved forest trees in vitro[J]. Commun Inst. For. Cech.,1981,12:255-271.
[8]袁巧平,董茂山,Christian Jay-Allemand.离体培养条件下核桃器官发生和体细胞胚胎发生[J].林业科学,1990,6:495-499,581-582.
[9]汤浩茹,王永清,任正隆.核桃体细胞胚发生与转基因研究进展[J].林业科学,2000,36(3):102-110.
[10]李好先.核桃遗传转化体系的建立及rolB基因的导入[D].北京:中国农业科学院,2013.
[11]张燕,范宏伟,张国强,等.汾阳核桃叶片愈伤组织的诱导、增殖及分化培养[J].中国农学通报,2006,22(8):91-91.
[12]Heile-Sudholt C,Huetteman C A,Preece J E,Van Sambeek J W,Gaffney G R,In vitro embryonic axis and seedling shoot tip culture of Juglans nigra L[J]. Plant Cell Tissue Organ Cult. 1986,(6):189–197.
[13]李好先,曹尚银,薛辉,等.‘绿波’核桃叶片愈伤组织的诱导研究[J].分子植物育种,2015,(5):1060-1066.