Departamento de Biodiversidad y Biología Experimental, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Intendente Guiraldes 2160, Pab. 2, Ciudad Universitaria, C1428EGA, Argentina.
Ann Bot. 2013 Nov;112(7):1253-62. doi: 10.1093/aob/mct184. Epub 2013 Sep 22.
In mature quinoa (Chenopodium quinoa) seeds, the lasting endosperm forms a micropylar cone covering the radicle. The suspensor cells lie within the centre of the cone. During the final stage of seed development, the cells of the lasting endosperm accumulate protein and lipids while the rest are crushed and disintegrated. Both the suspensor and endosperm die progressively from the innermost layers surrounding the embryo and extending towards the nucellar tissue. Ricinosomes are endoplasmic reticulum-derived organelles that accumulate both the pro-form and the mature form of cysteine endopeptidase (Cys-EP), first identified in castor bean (Ricinus communis) endosperm during germination. This study sought to identify associations between the presence of ricinosomes and programmed cell death (PCD) hallmarks in suspensor and endosperm cells predestined to die during quinoa seed development.
A structural study using light microscopy and transmission electron microscopy was performed. To detect the presence of Cys-EP, both western blot and in situ immunolocalization assays were carried out using anti-R. communis Cys-EP antibody. A TUNEL assay was used to determine DNA fragmentation.
Except for the one or two cell layers that constitute the lasting endosperm in the mature seed, ricinosomes were found in suspensor and endosperm cells. These cells were also the site of morphological abnormalities, including misshapen and fragmented nuclei, vesiculation of the cytosol, vacuole collapse and cell wall disorganization. It is proposed that, in suspensor and endosperm cells, the early detection of Cys-EP in ricinosomes predicts the occurrence of PCD during late seed development.
在成熟的藜麦(Chenopodium quinoa)种子中,持久胚乳形成一个覆盖胚根的珠孔锥形物。胚柄细胞位于锥形物的中心。在种子发育的最后阶段,持久胚乳的细胞积累蛋白质和脂质,而其余的则被压碎和分解。胚柄和胚乳都从最内层的胚胎周围逐渐死亡,并向珠心组织延伸。类蓖麻体是内质网衍生的细胞器,在蓖麻(Ricinus communis)胚乳在萌发过程中积累半胱氨酸内肽酶(Cys-EP)的原形式和成熟形式。本研究旨在鉴定类蓖麻体与在藜麦种子发育过程中注定死亡的胚柄和胚乳细胞程序性细胞死亡(PCD)特征之间的关联。
使用光镜和透射电镜进行结构研究。使用抗蓖麻 Cys-EP 抗体进行 Western blot 和原位免疫定位检测,以检测 Cys-EP 的存在。使用 TUNEL 测定法测定 DNA 片段化。
除了成熟种子中构成持久胚乳的一到两个细胞层外,在胚柄和胚乳细胞中都发现了类蓖麻体。这些细胞也是形态异常的部位,包括畸形和碎片化的核、细胞质泡状化、液泡塌陷和细胞壁紊乱。因此,在胚柄和胚乳细胞中,类蓖麻体中 Cys-EP 的早期检测预示着晚期种子发育过程中 PCD 的发生。