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首页> 外文期刊>Protoplasma: An International Journal of Cell Biology >A milestone in the doubled haploid pathway of cassava
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A milestone in the doubled haploid pathway of cassava

机译：木薯单倍体途径加倍的里程碑

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This study was aimed at inducing androgenesis in cultured anthers of cassava (Manihot esculenta Crantz) to develop a protocol for the production of doubled haploids. Microspore reprogramming was induced in cassava by cold or heat stress of anthers. Since the anthers contain both haploid microspores and diploid somatic cells, it was essential to verify the origin of anther-derived calli. The origin of antherderived calli was assessed bymorphological screening followed by histological analysis and flow cytometry (FCM). Additionally, simple sequence repeat (SSR) and amplified fragmented length polymorphism (AFLP) assays were used for the molecular identification of the microspore-derived calli. The study clearly demonstrated the feasibility of producing microspore-derived calli using heat- or cold-pretreated anthers. Histological studies revealed reprogramming of the developmental pathway of microspores by symmetrical division of the nucleus. Flow cytometry analysis revealed different ploidy level cell types including haploids, which confirmed their origin from the microspores. The SSR and AFLP marker assays independently confirmed the histological and FCMresults of a haploid origin of the calli at the DNA level. The presence of multicellular microspores in the in vitro system indicated a switch of developmental program, which constitutes a crucial step in the design of protocols for the regeneration of microspore-derived embryos and plants. This is the first detailed report of calli, embryos, and abnormal shoots originated from the haploid cells in cassava, leading to the development of a protocol for the production of doubled haploid plants in cassava.
机译：这项研究旨在诱导木薯培养的花药中的雄激素生成（Manihot esculenta Crantz），以开发生产双倍单倍体的方案。花药的冷或热胁迫在木薯中诱导了小孢子重编程。由于花药同时包含单倍体小孢子和二倍体体细胞，因此必须验证花药来源的愈伤组织的起源。通过形态学筛选，组织学分析和流式细胞术（FCM）评估花药愈伤组织的起源。此外，简单的序列重复（SSR）和扩增的片段长度多态性（AFLP）分析用于分子鉴定小孢子来源的愈伤组织。该研究清楚地证明了使用热或冷预处理的花药生产小孢子来源的愈伤组织的可行性。组织学研究显示，小孢子的发育途径通过细胞核的对称分裂而重新编程。流式细胞仪分析揭示了不同倍性水平的细胞类型，包括单倍体，证实了它们起源于小孢子。 SSR和AFLP标记测定法在DNA水平上独立证实了愈伤组织单倍体起源的组织学和FCM结果。体外系统中多细胞小孢子的存在表明发展计划的转变，这构成了设计用于再生小孢子来源的胚和植物的方案的关键步骤。这是关于木薯中单倍体细胞的愈伤组织，胚胎和异常芽的首次详细报道，从而导致开发出在木薯中生产双倍单倍体植物的方案。

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