标题：Sysmex Partec 倍性分析仪在植物种子研究上的应用---木薯原生质体电融合植株再生
Abstract：
Protoplast electrofusion between callus protoplasts of cultivar TMS60444 and mesophyll protoplasts of cultivar SC8 was performed as an approach for the genetic improvement of cassava. The fusion products were subsequently cultured in protoplast culture medium (TM2G) with gradual dilution for approximately 1–2 months. Then the protoplast-derived compact calli were transferred to suspension culture medium (SH) for suspension culture. The cultured products developed successively into embryos, mature embryos, and shoots on somatic embryo emerging medium (MSN), embryo maturation medium (CMM), and shoot elongation medium (CEM), respectively. And the shoots were then rooted on Murashige and Skoog (1962) medium (MS). Sixty-six cell lines were obtained and 12 of them developed into plantlets. Based on assessment of ploidy level and chromosome counting, four of these plantlets were tetraploid and the remaining eight were diploid. Based on assessment of ploidy level and simple sequence repeat (SSR) analysis, nine tetraploid cell lines, one diploid variant plant line and nine variant cell lines were obtained. The diploid variant plant line and the nine variant cell lines all showed partial loss of genetic material compared to that of the parent TMS60444, based on SSR patterns. These results showed that some new germplasm of cassava were created. In this study, a protocol for protoplast electrofusion was developed and validated. Another important conclusion from this work is the confirmation of a viable protocol for the regeneration of plants from cassava protoplasts. Going forward, we hope to provide technical guidance for cassava tissue culture, and
also provide some useful inspiration and reference for further genetic improvement of cassava.
摘要：
将TMS60444品种的愈伤组织原生质体与SC8品种的叶肉原生质体进行原生质体电融合，作为木薯遗传改良的一种方法。随后将融合产物在原生质体培养基（TM2G）中逐渐稀释培养约1-2个月。然后将原生质体衍生的紧密愈伤组织转移到悬浮培养基（SH）中进行悬浮培养。培养产物分别在体细胞胚胎发生培养基（MSN）、胚胎成熟培养基（CMM）和芽伸长培养基（CEM）上发育成胚胎、成熟胚胎和芽。然后将芽在Murashige和Skoog（1962）培养基（MS）上生根。获得了66个细胞系，其中12个发育成小植株。根据倍性水平和染色体计数的评估，其中4株为四倍体，其余8株为二倍体。基于倍性水平评估和简单序列重复（SSR）分析，获得了9个四倍体细胞系、1个二倍体变异植株系和9个变异细胞系。基于SSR图谱，与亲本TMS60444相比，二倍体变异植物系和九个变异细胞系都显示出部分遗传物质损失。这些结果表明，木薯创造了一些新的种质资源。在这项研究中，开发并验证了原生质体电融合的方案。这项工作的另一个重要结论是确认了木薯原生质体再生植物的可行方案。展望未来，我们希望为木薯组织培养提供技术指导，也为木薯的进一步遗传改良提供一些有益的启示和参考。
关键词：
Sysmex Partec 倍性分析仪，多倍体诱导仪，CyFlow Ploidy Analyser 流式细胞仪，组织培养、染色体计数、DNA丢失、ploidy analysis 倍性分析
