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Microspore reprogramming to embryogenesis Induces changes in cell wall and starch accumulation dynamics associated with proliferation and differentiation events

机译:小孢子重编程为胚胎发生诱导与增殖和分化事件相关的细胞壁和淀粉积累动态的变化

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摘要

Plant cell wall polymers are regulated during development, but the specific roles of their different molecular components and the functional meaning of cell wall changes in different cell types and cell processes are still unclear. In the present workthe presence and distribution of different cell wall components in Cops/cum annuum L pollen have been analyzed in situ in order to monitor how they change during two developmental programs. These programs are: pollen development, which is a differentiation process, and stress-induced pollen reprogramming to embryogenesis, which involves proliferation followed later by differentiation processes. Specific antibodies recognizing the major cell wall polymers, the major hemicellulose, xyloglucan (XG), the rhamnogalacturonan II (RGII) pectin domain and high- and low-methyl-esterified pectins were used for both dot-blot and immunolocalization assays at light and electron microscopy levels during defined developmental stages. For comparison purposes, a similar approach was also used in zygotic embryogenesis and root apical tip growth. Results showed differences in the distribution pattern of these molecular complexes, in the proportion of esterified and de-esterified pectins in the two pollen developmental pathways, and defined wall changes during microspore reprogramming. These changes were associated with proliferation and differentiation events where highly esterified pectins were characteristic of proliferation, while de-esterified pectins, XG and RGIIwere abundant in walls of differentiating cells. Starch deposits were also studied and the results revealed changes in starch synthesis dynamics after switching the pollen embryogenic developmental program. These changes occurred together with modifications in the distribution patterns of cell wall polymers, starch accumulation being associated with cell differentiation. As in the case of proliferating cells, esterified pectins were also abundant in the apertures of developing microspores, regions of new cell wall formation. The different distribution patterns of cell wall polymers were common for proliferating cells and differentiating cells in all the plant systems analyzed, including zygotic embryos and root tip cells, suggesting that these patternsare markers of proliferation and differentiation events as well as markers of pollen reprogramming to embryogenesis.
机译:植物细胞壁聚合物在发育过程中受到调节,但是它们不同分子成分的具体作用以及在不同细胞类型和细胞过程中细胞壁变化的功能含义仍不清楚。在目前的工作中,已经对原代Cops / cumulum L花粉中不同细胞壁成分的存在和分布进行了现场分析,以监测它们在两个发育程序中的变化。这些程序是:花粉发育,这是一个分化过程,以及应激诱导的花粉重编程为胚胎发生,其涉及增殖,随后是分化过程。识别主要细胞壁聚合物,主要半纤维素,木葡聚糖(XG),鼠李糖半乳糖醛酸聚糖II(RGII)果胶结构域以及高和低甲基酯化果胶的特异性抗体用于光和电子斑点印迹和免疫定位测定在确定的发育阶段的显微镜检查水平。为了进行比较,在合子胚发生和根尖生长中也使用了类似的方法。结果表明,这些分子复合物的分布方式,两种花粉发育途径中的酯化果胶和去酯化果胶的比例存在差异,并且在小孢子重编程过程中定义了壁的变化。这些变化与增殖和分化事件有关,其中高度酯化的果胶是增殖的特征,而去酯化的果胶,XG和RGII在分化细胞壁中丰富。还研究了淀粉沉积物,结果揭示了在切换花粉胚胎发生发育程序后淀粉合成动力学的变化。这些变化与细胞壁聚合物分布模式的改变一起发生,淀粉积累与细胞分化有关。与增殖细胞的情况一样,酯化果胶在发育中的小孢子的孔中也很丰富,这些孔是新细胞壁形成的区域。在所有分析的植物系统中,包括合子胚和根尖细胞,细胞壁聚合物的不同分布模式对于增殖细胞和分化细胞都是常见的,这表明这些模式是增殖和分化事件的标志物,以及花粉重编程为胚胎发生的标志物。

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