scholarly journals Proteomic Analysis of Generative and Vegetative Nuclei Reveals Molecular Characteristics of Pollen Cell Differentiation in Lily

2021 ◽  
Vol 12 ◽  
Author(s):  
Chen You ◽  
YuPing Zhang ◽  
ShaoYu Yang ◽  
Xu Wang ◽  
Wen Yao ◽  
...  

In plants, the cell fates of a vegetative cell (VC) and generative cell (GC) are determined after the asymmetric division of the haploid microspore. The VC exits the cell cycle and grows a pollen tube, while the GC undergoes further mitosis to produce two sperm cells for double fertilization. However, our understanding of the mechanisms underlying their fate differentiation remains limited. One major advantage of the nuclear proteome analysis is that it is the only method currently able to uncover the systemic differences between VC and GC due to GC being engulfed within the cytoplasm of VC, limiting the use of transcriptome. Here, we obtained pure preparations of the vegetative cell nuclei (VNs) and generative cell nuclei (GNs) from germinating lily pollens. Utilizing these high-purity VNs and GNs, we compared the differential nucleoproteins between them using state-of-the-art quantitative proteomic techniques. We identified 720 different amount proteins (DAPs) and grouped the results in 11 fate differentiation categories. Among them, we identified 29 transcription factors (TFs) and 10 cell fate determinants. Significant differences were found in the molecular activities of vegetative and reproductive nuclei. The TFs in VN mainly participate in pollen tube development. In comparison, the TFs in GN are mainly involved in cell differentiation and male gametogenesis. The identified novel TFs may play an important role in cell fate differentiation. Our data also indicate differences in nuclear pore complexes and epigenetic modifications: more nucleoporins synthesized in VN; more histone variants and chaperones; and structural maintenance of chromosome (SMC) proteins, chromatin remodelers, and DNA methylation-related proteins expressed in GN. The VC has active macromolecular metabolism and mRNA processing, while GC has active nucleic acid metabolism and translation. Moreover, the members of unfolded protein response (UPR) and programmed cell death accumulate in VN, and DNA damage repair is active in GN. Differences in the stress response of DAPs in VN vs. GN were also found. This study provides a further understanding of pollen cell differentiation mechanisms and also a sound basis for future studies of the molecular mechanisms behind cell fate differentiation.

2014 ◽  
Vol 50 (3) ◽  
pp. 367-380 ◽  
Author(s):  
Elżbieta Bednarska

DNA and histone synthesis in five consecutive morphological stages of <em>Hyacinthus orientalis</em> L. pollen grain differentiation were studied autoradiographically. DNA synthesis was found to occur in both the generative and the vegetative cell. DNA replication in the generative cell took place when the generative cell was still adhered to the pollen grain wall but already devoid of callose wall. DNA synthesis in the generative cell slightly preceded that in the vegetative cell. Histones were synthesized in phase S of the generative and vegetative cell. In the generative cell histone synthesis also continued at a lower level after completion of DNA replication. In the developmental stages under study the nuclei of the generative cells were decidedly richer in lysine histones than vegetative cell nuclei.


2020 ◽  
Vol 54 (1) ◽  
pp. 121-149 ◽  
Author(s):  
Benjamin Loppin ◽  
Frédéric Berger

Nucleosome dynamics and properties are central to all forms of genomic activities. Among the core histones, H3 variants play a pivotal role in modulating nucleosome structure and function. Here, we focus on the impact of H3 variants on various facets of development. The deposition of the replicative H3 variant following DNA replication is essential for the transmission of the epigenomic information encoded in posttranscriptional modifications. Through this process, replicative H3 maintains cell fate while, in contrast, the replacement H3.3 variant opposes cell differentiation during early embryogenesis. In later steps of development, H3.3 and specialized H3 variants are emerging as new, important regulators of terminal cell differentiation, including neurons and gametes. The specific pathways that regulate the dynamics of the deposition of H3.3 are paramount during reprogramming events that drive zygotic activation and the initiation of a new cycle of development.


Genome ◽  
2003 ◽  
Vol 46 (1) ◽  
pp. 83-88 ◽  
Author(s):  
Roberto Gargione Junqueira Filho ◽  
Andréa Beatriz Mendes-Bonato ◽  
Maria Suely Pagliarini ◽  
Nilton Cesar Pires Bione ◽  
Cacilda Borges do Valle ◽  
...  

Meiotic division and male gametophyte development were analyzed in one tetraploid (2n = 4x = 36) accession of Brachiaria decumbens cv. Basilisk that showed some pollen sterility. Meiotic process was typical of polyploids in that it consisted of multiple chromosome associations. Precocious chromosome migration to the poles, laggards, and micronucleus formation were abundant in both meiosis I and II and resulted in tetrads with micronuclei. After callose dissolution, microspores were released into the anther locule and had the semblance of being normal. Although each microspore initiated its differentiation by pollen mitosis, in 43.24% of the microspores, nuclear polarization was not observed and the typical hemispherical cell plate was not detected. Division was symmetric and microspores lacked differentiation between the vegetative and the generative cell. Both nuclei were of equal size, presented equal chromatin condensation, and had a spherical shape. After the first pollen mitosis and cytokinesis, each cell underwent a new symmetric mitosis without nuclear polarization. At the end of the second pollen mitosis, four equal nuclei were observed in each pollen grain. After the second cytokinesis, the cells gave rise to four equal-sized pollen grains with a similar tetrad configuration that initially remained together. Sterile pollen grains resulted from abnormal pollen mitosis. This anomaly may be explained by a mutation, probably affecting microtubule cytoskeleton formation. The importance of this male-sterile mutation for Brachiaria breeding programs is discussed.Key words: Brachiaria decumbens, male sterility, pollen mitosis, microspore polarity, symmetric division, pollen cell fate, grasses.


MRS Advances ◽  
2019 ◽  
Vol 5 (12-13) ◽  
pp. 601-607
Author(s):  
Jie Zou ◽  
Weiwei Wang ◽  
Xianlei Sun ◽  
Wingtai Tung ◽  
Nan Ma ◽  
...  

ABSTRACTThe dynamic mechanical force transmitted through microenvironments during tissue formation and regeneration continuously impacts the mechanics of cells and thereby regulates gene and protein expression. The mechanical properties are altered during the process of stem cells differentiating into different lineages. At different stages of differentiation, stem cells display different mechanical properties in response to surrounding microenvironments, which depend on the subcellular structures, especially the cytoskeleton and nucleus. The mechanical properties of the cell nucleus affect protein folding and transport as well as the condensation of chromatin, through which the cell fate is regulated. These findings raise the question as to how cell mechanics change during differentiation. In this study, the mechanical properties of human bone marrow mesenchymal stem cells (hBMSCs) were determined during adipogenic and osteogenic differentiation by atomic force microscopy (AFM). The cytoskeletal structure and the modification of histone were investigated using laser confocal microscope and flow cytometry. The mechanical properties of cell nuclei at different stages of cell differentiation were compared. The stiffness of nuclei increased with time as osteogenesis was induced in hBMSCs. The H3K27me3 level increased during osteogenesis and adipogenesis according to flow cytometry analysis. Our results show conclusively that AFM is a facile and effective method to monitor stem cell differentiation. The measurement of cell mechanical properties by AFM improves our understanding on the connection between mechanics and stem cell fate.


1977 ◽  
Vol 73 (2) ◽  
pp. 521-526 ◽  
Author(s):  
V Raghavan

Continued DNA synthesis in the generative cell nucleus, followed by mitosis and cytokinesis, results in the formation of pollen embryoids in cultured anthers of H. niger. In contrast, the nucleus of the vegetative cell undergoes no DNA synthesis after it is cut off, or synthesizes DNA only during a limited number of cell cycles. DNA synthetic patterns in the generative and vegetative cell nuclei confirm the ontogeny of embryoids described in this plant.


The development of the male gametophyte of Taxus baccata has been studied over a period of 20 weeks, from germination of the microspore in February to spermatogenesis in July. A few days after germination the microspore nucleus divides and a transverse wall forms at the equator cutting off the small generative cell and a large tube cell. The latter immediately begins to expand to form the pollen tube. The first division thus establishes the polarity of the gametophyte and the generative cell is regarded as proximal. The transverse wall is ephemeral, and within six weeks it has disappeared. The nucleus of the generative cell divides while still at the proximal pole. The two daughter nuclei are unequal in size, but they remain associated and together move distally. The larger nucleus eventually becomes the nucleus of the spermatogenous cell, and the smaller the sterile nucleus. The spermatogenous cell acquires a distinctive cytoplasm and becomes surrounded by a wall which arises de novo . The nucleus of the spermatogenous cell enlarges, but always remains towards one side of the cell so that at mitosis the spindle is contained within one hemisphere. After division the wall of the spermatogenous cell is ruptured and the two sperms are released as naked nuclei of equal size. The cytoplasm of the spermatogenous cell degenerates as it enters the tube, but remains recognizable until fertilization.


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