辣椒小孢子衍生多细胞的胚胎发生[j]。

Fan Liu, Hong Zhao, Bin Chen, Yue Yun Zhang
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引用次数: 0

摘要

将花药在固化培养基上预培养15d后,分离出小孢子和衍生的多细胞,在改性CP液体培养基中培养。悬浮培养30天后,在28℃的黑暗条件下形成不同发育阶段的胚状体。12个花药的细胞悬浮液可形成22个胚状体,其中约23%的胚状体处于子叶期。荧光和光镜观察显示这些胚状体来源于小孢子。单核小孢子的细胞核经过多次对称分裂,形成多核细胞或多细胞,并进一步发育成胚状体。TTC染色显示,辣椒胚状体表面有白色茸毛,部分胚状体活力较低,部分胚状体活力正常。鱼雷期和子叶期胚状体在凝固培养基上可形成植株。7℃、32℃和35℃胁迫均可诱导花药形成胚状体,7℃和35℃条件下花药胚发生频率较高,35℃和32℃条件下多细胞胚发生频率较高。讨论了产生这一结果的原因。不同基因型和不同温度胁迫条件下的胚发生频率存在明显差异。流式细胞分析结果显示,再生植株存在单倍体、双倍单倍体和单倍体-二倍体嵌合体。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
[Embryogenesis of microspore derived multicells in Capsicum annuum L].

Microspores and derived multicells were isolated and cultured in modified liquid CP medium after a 15d's preculture of anthers on solidified medium. Thirty days later in suspension culture, at 28 degrees C dark condition embryoids with different developmental stages were formed. Up to 22 embryoids could be formed from the cell suspension of 12 anthers, and about 23% of the embryoids were at the cotyledonary stage. Fluorescence and light microscope observations revealed that these embryoids derived from microspores. After several symmetrical division of the nuclei of uninucleated microspores, multi-nuclei cells or multi-cells were formed, and developed further into embryoids. There were white hairs on the surface of pepper embryoids, and some embryoids showed low vigor while others showed normal by TTC staining. Plants could be formed from torpedo and cotyledonary stage embryoids on solidified medium. Embryoids could be induced by 7 degrees C, 32 degrees C or 35 degrees C stress treatment on anthers, Higher embryogenesis frequencies were got at 7 degrees C and 35 degrees C condition in anther culture while 35 degrees C and 32 degrees C treatment showed a higher embryogenesis in isolated multicell culture. The reason of this result was discussed. There were obvious differences in embryogenesis frequency among different genotypes and different temperature stress conditions. Flow cytometric analysis revealed that there were haploidy, doubled haploidy and haploid-diploid chimera in the regenerated plants.

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