多基因型黑果枸杞高效快繁体系的建立

Establishment of Highly Efficient and Rapid Propagation System of Lycium ruthenicum for Multiple Genotypes

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.