Bae Jinjoo, Choi Yunseo, Song Jae-Young, Lee Jung-Ro, Yoon Munsup, Lee Young-Yi
National Agrobiodiversity Center, National Institute of Agricultural Sciences, RDA, Suwon 16613, Republic of Korea.
Department of Horticultural Bioscience, College of Natural Resources and Life Science, Pusan National University, Busan 50463, Republic of Korea.
Biology (Basel). 2022 Nov 30;11(12):1746. doi: 10.3390/biology11121746.
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.
为了长期保存遗传资源，已经开发了用于草莓种质的冷冻保存技术，主要使用离体培养的茎尖。在本研究中，在温室条件下对六个草莓种质（IT232511、PHS0132、IT245810、IT245830、IT245852和IT245860）进行了遗传稳定性测试，这些种质来源于以下程序：（1）常规繁殖（GH：温室保存）；（2）离体繁殖（TC：组织培养）；（3）冷冻保存前的预处理（-LN：未暴露于液氮）；（4）冷冻保存（+LN：暴露于液氮）。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，但三个种质的一些性状，如果实的含糖量和pH值，与GH相比，+LN中的值更高。然而，在第一代匍匐茎中差异消失。为了测试遗传变异，十二个简单序列重复区间（ISSR）引物共产生了102条带。仅在未进行冷冻保存的IT245860的TC衍生植株中发现了一些多态性条带。对ISSR_15产生的四个多态性条带的测序分析表明，这些序列在NCBI中均与已鉴定的基因不匹配。在所有植株中均未观察到表型异常。本研究表明，六个草莓种质的冷冻保存植株在表型和遗传上是稳定的。因此，本研究结果有助于实施草莓种质的低温保存库。
