標題：Sysmex Partec 倍性分析儀在植物種子研究上的應用---木薯原生質體電融合植株再生
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：
將TMS60444品種的愈傷組織原生質體與SC8品種的葉肉原生質體進行原生質體電融合，作為木薯遺傳改良的一種方法。隨后將融合產物在原生質體培養基（TM2G）中逐漸稀釋培養約1-2個月。然后將原生質體衍生的緊密愈傷組織轉移到懸浮培養基（SH）中進行懸浮培養。培養產物分別在體細胞胚胎發生培養基（MSN）、胚胎成熟培養基（CMM）和芽伸長培養基（CEM）上發育成胚胎、成熟胚胎和芽。然后將芽在Murashige和Skoog（1962）培養基（MS）上生根。獲得了66個細胞系，其中12個發育成小植株。根據倍性水平和染色體計數的評估，其中4株為四倍體，其余8株為二倍體。基于倍性水平評估和簡單序列重復（SSR）分析，獲得了9個四倍體細胞系、1個二倍體變異植株系和9個變異細胞系。基于SSR圖譜，與親本TMS60444相比，二倍體變異植物系和九個變異細胞系都顯示出部分遺傳物質損失。這些結果表明，木薯創造了一些新的種質資源。在這項研究中，開發并驗證了原生質體電融合的方案。這項工作的另一個重要結論是確認了木薯原生質體再生植物的可行方案。展望未來，我們希望為木薯組織培養提供技術指導，也為木薯的進一步遺傳改良提供一些有益的啟示和參考。
關鍵詞：
Sysmex Partec 倍性分析儀，多倍體誘導儀，CyFlow Ploidy Analyser 流式細胞儀，組織培養、染色體計數、DNA丟失、ploidy analysis 倍性分析
