为了长期保存遗传资源,已经开发了草莓种质的冷冻保存技术,主要使用体外生长的茎尖。在这项研究中,在温室条件下测试了六种草莓种质(IT232511,PHS0132,IT245810,IT245830,IT245852和IT245860)的遗传稳定性,这些草莓种质来自以下程序:(1)常规繁殖(GH:温室维持);(2)体外繁殖(TC:组织培养);(3)冷冻保存前的预处理(-LN:非液氮暴露);氮(4)为了测试表型性状的表现,我们测量了六个营养性状和五个果实性状。大多数特征没有明显差异,但是有一些特征,例如三个种质中水果的糖含量和pH值,与GH相比,+LN显示更高的值。然而,差异在第一代跑步者中消失了。为了测试遗传变异,通过12个简单序列间重复(ISSR)引物共产生102条条带。仅在源自IT245860的TC的植物中发现了一些多态性带,该植物未被冷冻保存。对ISSR_15产生的四个多态性条带的测序分析表明,这些序列均未与NCBI中的特征基因匹配。在所有植物中未观察到表型异常。这项研究表明,六个草莓种质的冷冻保存植物在表型和遗传上都是稳定的。因此,本研究结果有助于草莓种质资源的冷冻保存。
For the long-term preservation of genetic resources, cryopreservation techniques have been developed for strawberry germplasm, mainly using in vitro-grown shoot tips. In this study, genetic stability was tested under greenhouse conditions for six strawberry accessions (IT232511, PHS0132, IT245810, IT245830, IT245852, and IT245860) derived from the following procedures: (1) conventional propagation (GH: greenhouse maintained); (2) in vitro propagation (TC: tissue culture); (3) pretreatment before cryopreservation (-LN: non-liquid nitrogen exposure); and (4) cryopreservation (+LN: liquid nitrogen exposure). To test the performance of phenotypic traits, we measured six vegetative and five fruit traits. There were no distinct differences in most of the characteristics, but a few traits, such as sugar content and pH of fruits in three accessions, showed higher values in +LN compared to GH. However, the differences disappeared in the first runner generation. To test genetic variations, a total of 102 bands were generated by twelve inter simple sequence repeat (ISSR) primers. A few polymorphic bands were found only in plants derived from TC of IT245860, which was not cryopreserved. The sequencing analysis of four polymorphic bands produced by ISSR_15 showed that none of these sequences matched the characterized genes in NCBI. Phenotypic abnormality was not observed across all plants. This study indicates that cryopreserved plants of the six strawberry accessions are phenotypically and genetically stable. Therefore, the results of this study can help to implement cryobanking of strawberry germplasm.