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Unravelling the proteomic profile of rice meiocytes during early meiosis.

Collado-Romero M, Alós E, Prieto P - Front Plant Sci (2014)

Bottom Line: Moreover, the putative roles of 16 proteins which have not been previously associated to meiosis or were not identified in rice before, are also discussed namely: seven proteins involved in chromosome structure and remodeling, five regulatory proteins [such as SKP1 (OSK), a putative CDK2 like effector], a protein with RNA recognition motifs, a neddylation-related protein, and two microtubule-related proteins.Revealing the proteins involved in early meiotic processes could provide a valuable tool kit to manipulate chromosome associations during meiosis in rice breeding programs.The data have been deposited to the ProteomeXchange with the PXD001058 identifier.

View Article: PubMed Central - PubMed

Affiliation: Plant Breeding Department, Institute for Sustainable Agriculture, Agencia Estatal Consejo Superior de Investigaciones Científicas Córdoba, Spain.

ABSTRACT
Transfer of genetic traits from wild or related species into cultivated rice is nowadays an important aim in rice breeding. Breeders use genetic crosses to introduce desirable genes from exotic germplasms into cultivated rice varieties. However, in many hybrids there is only a low level of pairing (if existing) and recombination at early meiosis between cultivated rice and wild relative chromosomes. With the objective of getting deeper into the knowledge of the proteins involved in early meiosis, when chromosomes associate correctly in pairs and recombine, the proteome of isolated rice meiocytes has been characterized by nLC-MS/MS at every stage of early meiosis (prophase I). Up to 1316 different proteins have been identified in rice isolated meiocytes in early meiosis, being 422 exclusively identified in early prophase I (leptotene, zygotene, or pachytene). The classification of proteins in functional groups showed that 167 were related to chromatin structure and remodeling, nucleic acid binding, cell-cycle regulation, and cytoskeleton. Moreover, the putative roles of 16 proteins which have not been previously associated to meiosis or were not identified in rice before, are also discussed namely: seven proteins involved in chromosome structure and remodeling, five regulatory proteins [such as SKP1 (OSK), a putative CDK2 like effector], a protein with RNA recognition motifs, a neddylation-related protein, and two microtubule-related proteins. Revealing the proteins involved in early meiotic processes could provide a valuable tool kit to manipulate chromosome associations during meiosis in rice breeding programs. The data have been deposited to the ProteomeXchange with the PXD001058 identifier.

No MeSH data available.


Flow chart overview for the extraction of rice meiocytes in prophase I, the subsequent proteomics experiments and the data analysis.
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Figure 1: Flow chart overview for the extraction of rice meiocytes in prophase I, the subsequent proteomics experiments and the data analysis.

Mentions: Ethanol and acetic acid were used as fixatives because they have been previously proved to be successfully employed in proteomic analysis (Ahram et al., 2003; De Souza et al., 2004; Milcheva et al., 2013) (Figure 1). Although the anther length is generally used as a criterion to determine the developmental stages of anthers in rice (Kerim et al., 2003; Itoh et al., 2005; Nonomura et al., 2011), an anther from each fixed flower was removed using fine forceps under a dissection microscope (Stemi 2000-C stereomicroscope, Carl Zeiss, Göttingen, Germany) equipped with a cold-light source to stage meiosis as much accurately as possible. Anthers were then stained in acetocarmine solution, squashed on ethanol-cleaned slides and checked under a PrimoStar light microscope (Carl Zeiss, Göttingen, Germany). The identification of the meiotic developmental stage was based on previous cytological descriptions of rice male meiosis (Chen et al., 2005; Itoh et al., 2005). Photographs were taken using an AxioCam ICc3 digital camera (Carl Zeiss, Göttingen, Germany) attached to the microscope.


Unravelling the proteomic profile of rice meiocytes during early meiosis.

Collado-Romero M, Alós E, Prieto P - Front Plant Sci (2014)

Flow chart overview for the extraction of rice meiocytes in prophase I, the subsequent proteomics experiments and the data analysis.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4109522&req=5

Figure 1: Flow chart overview for the extraction of rice meiocytes in prophase I, the subsequent proteomics experiments and the data analysis.
Mentions: Ethanol and acetic acid were used as fixatives because they have been previously proved to be successfully employed in proteomic analysis (Ahram et al., 2003; De Souza et al., 2004; Milcheva et al., 2013) (Figure 1). Although the anther length is generally used as a criterion to determine the developmental stages of anthers in rice (Kerim et al., 2003; Itoh et al., 2005; Nonomura et al., 2011), an anther from each fixed flower was removed using fine forceps under a dissection microscope (Stemi 2000-C stereomicroscope, Carl Zeiss, Göttingen, Germany) equipped with a cold-light source to stage meiosis as much accurately as possible. Anthers were then stained in acetocarmine solution, squashed on ethanol-cleaned slides and checked under a PrimoStar light microscope (Carl Zeiss, Göttingen, Germany). The identification of the meiotic developmental stage was based on previous cytological descriptions of rice male meiosis (Chen et al., 2005; Itoh et al., 2005). Photographs were taken using an AxioCam ICc3 digital camera (Carl Zeiss, Göttingen, Germany) attached to the microscope.

Bottom Line: Moreover, the putative roles of 16 proteins which have not been previously associated to meiosis or were not identified in rice before, are also discussed namely: seven proteins involved in chromosome structure and remodeling, five regulatory proteins [such as SKP1 (OSK), a putative CDK2 like effector], a protein with RNA recognition motifs, a neddylation-related protein, and two microtubule-related proteins.Revealing the proteins involved in early meiotic processes could provide a valuable tool kit to manipulate chromosome associations during meiosis in rice breeding programs.The data have been deposited to the ProteomeXchange with the PXD001058 identifier.

View Article: PubMed Central - PubMed

Affiliation: Plant Breeding Department, Institute for Sustainable Agriculture, Agencia Estatal Consejo Superior de Investigaciones Científicas Córdoba, Spain.

ABSTRACT
Transfer of genetic traits from wild or related species into cultivated rice is nowadays an important aim in rice breeding. Breeders use genetic crosses to introduce desirable genes from exotic germplasms into cultivated rice varieties. However, in many hybrids there is only a low level of pairing (if existing) and recombination at early meiosis between cultivated rice and wild relative chromosomes. With the objective of getting deeper into the knowledge of the proteins involved in early meiosis, when chromosomes associate correctly in pairs and recombine, the proteome of isolated rice meiocytes has been characterized by nLC-MS/MS at every stage of early meiosis (prophase I). Up to 1316 different proteins have been identified in rice isolated meiocytes in early meiosis, being 422 exclusively identified in early prophase I (leptotene, zygotene, or pachytene). The classification of proteins in functional groups showed that 167 were related to chromatin structure and remodeling, nucleic acid binding, cell-cycle regulation, and cytoskeleton. Moreover, the putative roles of 16 proteins which have not been previously associated to meiosis or were not identified in rice before, are also discussed namely: seven proteins involved in chromosome structure and remodeling, five regulatory proteins [such as SKP1 (OSK), a putative CDK2 like effector], a protein with RNA recognition motifs, a neddylation-related protein, and two microtubule-related proteins. Revealing the proteins involved in early meiotic processes could provide a valuable tool kit to manipulate chromosome associations during meiosis in rice breeding programs. The data have been deposited to the ProteomeXchange with the PXD001058 identifier.

No MeSH data available.