多基因型黑果枸杞高效快繁体系的建立
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摘要
旨为建立适用于不同种源多基因型的黑果枸杞高效稳定的组培再生体系,为工厂化育苗与大面积推广奠定基础。收集7个不同种源地的黑果枸杞种子并制备无菌苗,利用植物组织培养技术,研究不同种类、不同浓度植物生长调节剂对黑果枸杞生根和分化的影响。在黑果枸杞的茎段诱导分化培养基筛选过程中,针对黑果枸杞培养过程中容易出现的玻璃化现象,通过多种培养基比较,确定了一种最适合不同种源多基因型黑果枸杞茎段分化的培养基MS+蔗糖30 g/L+琼脂7 g/L+6-BA 0.1 mg/L;在黑果枸杞的叶片诱导分化培养基筛选过程中,通过调节生长素类物质及细胞分裂素的浓度,获得一种适合于不同基因型的黑果枸杞叶片分化的培养基MS+蔗糖30 g/L+琼脂6 g/L+NAA 0.5 mg/L+6-BA 0.5 mg/L;在黑果枸杞生根培养基的筛选过程中,通过改变生长素浓度,获得了一种最适合于不同种源多基因型黑果枸杞生根的培养基1/2MS+蔗糖30 g/L+琼脂7.5 g/L+IBA0.25 mg/L。在大规模试验的基础上,克服了不同种源黑果枸杞使用一种培养基时在生根分化过程中常出现的玻璃化、不生根、不发芽等问题,建立了适宜不同种源多基因型黑果枸杞高效稳定的组织培养再生体系。
This work aims to establish an efficient and stable tissue culture regeneration system suitable for different provenances and multiple genotypes of Lycium ruthenicum Murr. and lay a foundation for planting-raising and large-scale popularization. The seeds of L.ruthenicum from 7 different provenances were collected and sterile seedlings were prepared. Plant tissue culture techniques were used to study the effects of different kinds and concentrations of plant growth regulators on the rooting and differentiation of L. ruthenicum. In the process of screening stem segment differentiation medium of L. ruthenicum,aiming at the vitrification phenomenon which is easy to appear in the culture process of L. ruthenicum,a medium suitable for the stem differentiations of multi-provenance and multi-genotype L. ruthenicum was determined through comparison of various media as such :MS + sucrose 30 g/L + agar 7 g/L + 6-BA 0.1 mg/L. A medium suitable for the leaf differentiation of multi-genotype L. ruthenicum was obtained as MS + sucrose 30 g/L + agar 6 g/L + NAA 0.5 mg/L + 6-BA 0.5 mg/L by adjusting the concentration of auxin and cell division while screening leaf-induced differentiation medium of L. ruthenicum. An optimal medium 1/2 MS+ sucrose 30 g/L+ agar 7.5 g/L + IBA0.25 mg/L for the rooting of and multi-provenance and multi-genotype L. ruthenicum was determined by changing the concentration of auxin. Based on the large-scale experiments,the problems of vitrification,non-rooting,non-germination,etc.,which often occur during rooting and differentiation of Lycium ruthenicum from different provenances when using one medium,were overcome.In another word,an efficient and stable tissue culture and regeneration system suitable for multi-provenance and multi-genotype L. ruthenicum was established.
This work aims to establish an efficient and stable tissue culture regeneration system suitable for different provenances and multiple genotypes of Lycium ruthenicum Murr. and lay a foundation for planting-raising and large-scale popularization. The seeds of L.ruthenicum from 7 different provenances were collected and sterile seedlings were prepared. Plant tissue culture techniques were used to study the effects of different kinds and concentrations of plant growth regulators on the rooting and differentiation of L. ruthenicum. In the process of screening stem segment differentiation medium of L. ruthenicum,aiming at the vitrification phenomenon which is easy to appear in the culture process of L. ruthenicum,a medium suitable for the stem differentiations of multi-provenance and multi-genotype L. ruthenicum was determined through comparison of various media as such :MS + sucrose 30 g/L + agar 7 g/L + 6-BA 0.1 mg/L. A medium suitable for the leaf differentiation of multi-genotype L. ruthenicum was obtained as MS + sucrose 30 g/L + agar 6 g/L + NAA 0.5 mg/L + 6-BA 0.5 mg/L by adjusting the concentration of auxin and cell division while screening leaf-induced differentiation medium of L. ruthenicum. An optimal medium 1/2 MS+ sucrose 30 g/L+ agar 7.5 g/L + IBA0.25 mg/L for the rooting of and multi-provenance and multi-genotype L. ruthenicum was determined by changing the concentration of auxin. Based on the large-scale experiments,the problems of vitrification,non-rooting,non-germination,etc.,which often occur during rooting and differentiation of Lycium ruthenicum from different provenances when using one medium,were overcome.In another word,an efficient and stable tissue culture and regeneration system suitable for multi-provenance and multi-genotype L. ruthenicum was established.
引文
[1]Chen SS,Zeng Z,Hu N,et al.Simultaneous optimization of the ultrasound-assisted extraction for phenolic compounds content and antioxidant activity of Lycium ruthenicum Murr.fruit using response surface methodology[J].Food Chemistry,2018,242(2018):1-8.
[2]李进,瞿伟菁.大孔树脂吸附分离黑果枸杞色素的研究[J].食品科学,2005,26(6):47-51.
[3]陈晨,文怀秀,赵晓辉,等.黑果枸杞色素中原花青素含量的测定[J].光谱实验室,2011,28(4):1767-1769.
[4]杭园园,李悦,王朝警,等.黑果枸杞花青素类型、含量及结构分析研究[J].食品研究与开发,2018,39(13):143-148.
[5]Liu Y,Song Y,Zeng S,et al.Isolation and characterization of a salt stress-responsive betaine aldehyde dehydrogenase in Lycium ruthenicum Murr.[J].Physiologia Plantarum,2018,163(1):73-87.
[6]《内蒙古植物志》编辑委员会.内蒙古植物志[M]:2版.呼和浩特:内蒙古人民出版社,1998:230.
[7]中国科学院西北高原生物研究所.青海植物志[M].西宁:青海省人民出版社,1999:176.
[8]马德滋,刘惠兰.宁夏植物志[M]:2版.银川:宁夏人民出版社,1986:156-157.
[9]米吉提·胡达拜尔地,潘晓巧.新疆植物志[M]:4版.乌鲁木齐:新疆科学技术出版社,2004
[10]汪智军,靳开颜,古丽森.新疆枸杞属植物资源调查及其保育措施[J].北方园艺,2013(3):169-171.
[11]白春雷,王灵茂,王志国,等.黑果枸杞育苗技术综述[J].种子科技,2016,34(12):51-54.
[12]马金平,李建国,王孝,等.黑果枸杞苗木快速繁育及建园技术[J].北方园艺,2013(9):185-187.
[13]唐菊秀.黑果枸杞的特性及育苗技术[J].农业科技与信息,2014(19):9-11.
[14]浩仁塔本,赵颖,郭永盛,等.黑果枸杞的组织培养[J].植物生理学报,2005,41(5):631-631.
[15]包振华,郭军战,周玮,等.枸杞组织培养再生体系优化[J].西北林学院学报,2010,25(5):73-76.
[16]杜敏智.大果黑果枸杞组培快繁技术体系的研究[D].延吉:延边大学,2015.
[17]孙晓红,位书磊,宋强,等.黑果枸杞的叶片分化与快速繁殖[J].植物生理学报,2016,52(5):653-658.
[18]魏艳,赵惠恩.百里香组织培养中克服玻璃化现象的初探[J].内蒙古农业大学学报:自然科学版,2007,28(2):205-208.
[19]王方琳,柴成武,魏小红,等.荒漠区药用植物黑果枸杞(Lycium ruthencium)的组织培养[J].干旱区资源与环境,2016,30(10):104-109.
[20]Sun SY,Cao HN,Yao H.Study on tissue culture and rapid propagation of Lycium ruthenicum Murr.[J].Agricultural Science&Technology,2016,17(5):1060-1064.
[2]李进,瞿伟菁.大孔树脂吸附分离黑果枸杞色素的研究[J].食品科学,2005,26(6):47-51.
[3]陈晨,文怀秀,赵晓辉,等.黑果枸杞色素中原花青素含量的测定[J].光谱实验室,2011,28(4):1767-1769.
[4]杭园园,李悦,王朝警,等.黑果枸杞花青素类型、含量及结构分析研究[J].食品研究与开发,2018,39(13):143-148.
[5]Liu Y,Song Y,Zeng S,et al.Isolation and characterization of a salt stress-responsive betaine aldehyde dehydrogenase in Lycium ruthenicum Murr.[J].Physiologia Plantarum,2018,163(1):73-87.
[6]《内蒙古植物志》编辑委员会.内蒙古植物志[M]:2版.呼和浩特:内蒙古人民出版社,1998:230.
[7]中国科学院西北高原生物研究所.青海植物志[M].西宁:青海省人民出版社,1999:176.
[8]马德滋,刘惠兰.宁夏植物志[M]:2版.银川:宁夏人民出版社,1986:156-157.
[9]米吉提·胡达拜尔地,潘晓巧.新疆植物志[M]:4版.乌鲁木齐:新疆科学技术出版社,2004
[10]汪智军,靳开颜,古丽森.新疆枸杞属植物资源调查及其保育措施[J].北方园艺,2013(3):169-171.
[11]白春雷,王灵茂,王志国,等.黑果枸杞育苗技术综述[J].种子科技,2016,34(12):51-54.
[12]马金平,李建国,王孝,等.黑果枸杞苗木快速繁育及建园技术[J].北方园艺,2013(9):185-187.
[13]唐菊秀.黑果枸杞的特性及育苗技术[J].农业科技与信息,2014(19):9-11.
[14]浩仁塔本,赵颖,郭永盛,等.黑果枸杞的组织培养[J].植物生理学报,2005,41(5):631-631.
[15]包振华,郭军战,周玮,等.枸杞组织培养再生体系优化[J].西北林学院学报,2010,25(5):73-76.
[16]杜敏智.大果黑果枸杞组培快繁技术体系的研究[D].延吉:延边大学,2015.
[17]孙晓红,位书磊,宋强,等.黑果枸杞的叶片分化与快速繁殖[J].植物生理学报,2016,52(5):653-658.
[18]魏艳,赵惠恩.百里香组织培养中克服玻璃化现象的初探[J].内蒙古农业大学学报:自然科学版,2007,28(2):205-208.
[19]王方琳,柴成武,魏小红,等.荒漠区药用植物黑果枸杞(Lycium ruthencium)的组织培养[J].干旱区资源与环境,2016,30(10):104-109.
[20]Sun SY,Cao HN,Yao H.Study on tissue culture and rapid propagation of Lycium ruthenicum Murr.[J].Agricultural Science&Technology,2016,17(5):1060-1064.