Limits...
Aberrant Meiotic Prophase I Leads to Genic Male Sterility in the Novel TE5A Mutant of Brassica napus

View Article: PubMed Central - PubMed

ABSTRACT

Genic male sterility (GMS) has already been extensively utilized for hybrid rapeseed production. TE5A is a novel thermo-sensitive dominant GMS line in Brassica napus, however, its mechanisms of GMS remain largely unclear. Histological and Transmission electron microscopy (TEM) analyses of anthers showed that the male gamete development of TE5A was arrested at meiosis prophase I. EdU uptake of S-phase meiocytes revealed that the TE5A mutant could accomplish DNA replication, however, chromosomal and fluorescence in situ hybridization (FISH) analyses of TE5A showed that homologous chromosomes could not pair, synapse, condense and form bivalents. We then analyzed the transcriptome differences between young floral buds of sterile plants and its near-isogenic fertile plants through RNA-Seq. A total of 3,841 differentially expressed genes (DEGs) were obtained, some of which were associated with homologous chromosome behavior and cell cycle control during meiosis. Dynamic expression changes of selected candidate DEGs were then analyzed at different anther developmental stages. The present study not only demonstrated that the TE5A mutant had defects in meiotic prophase I via detailed cytological analysis, but also provided a global insight into GMS-associated DEGs and elucidated the mechanisms of GMS in TE5A through RNA-Seq.

No MeSH data available.


Confocal images of EdU labeled meiocytes.EdU labeling of DNA is detected by a green fluorescent signal. DNA was counterstained with DAPI (blue). EdU uptake of S-phase meiocytes revealed that TE5A mutant could succeed in completing DNA replication (A–C) compared with the wild-type (D–F). Scale bars = 5 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC5037387&req=5

f6: Confocal images of EdU labeled meiocytes.EdU labeling of DNA is detected by a green fluorescent signal. DNA was counterstained with DAPI (blue). EdU uptake of S-phase meiocytes revealed that TE5A mutant could succeed in completing DNA replication (A–C) compared with the wild-type (D–F). Scale bars = 5 μm.

Mentions: Recently a new thymidine analog 5-ethynyl-2-deoxyuridine (EdU), was developed and utilized in investigating meiotic DNA replication. DNA is labeled through EdU uptake by S-phase meiocytes undergoing DNA replication. EdU labeling of DNA is detected with a fluorophore-tagged azide that forms a covalent bond with the terminal acetylene group on the ethynyl component of EdU. This new technology has been extensively employed in animals to study cell proliferation414243. EdU uptake of S-phase meiocytes revealed that TE5A mutants could succeed in completing DNA replication (Fig. 6A–C) compared to the wild-type (Fig. 6D–F).


Aberrant Meiotic Prophase I Leads to Genic Male Sterility in the Novel TE5A Mutant of Brassica napus
Confocal images of EdU labeled meiocytes.EdU labeling of DNA is detected by a green fluorescent signal. DNA was counterstained with DAPI (blue). EdU uptake of S-phase meiocytes revealed that TE5A mutant could succeed in completing DNA replication (A–C) compared with the wild-type (D–F). Scale bars = 5 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5037387&req=5

f6: Confocal images of EdU labeled meiocytes.EdU labeling of DNA is detected by a green fluorescent signal. DNA was counterstained with DAPI (blue). EdU uptake of S-phase meiocytes revealed that TE5A mutant could succeed in completing DNA replication (A–C) compared with the wild-type (D–F). Scale bars = 5 μm.
Mentions: Recently a new thymidine analog 5-ethynyl-2-deoxyuridine (EdU), was developed and utilized in investigating meiotic DNA replication. DNA is labeled through EdU uptake by S-phase meiocytes undergoing DNA replication. EdU labeling of DNA is detected with a fluorophore-tagged azide that forms a covalent bond with the terminal acetylene group on the ethynyl component of EdU. This new technology has been extensively employed in animals to study cell proliferation414243. EdU uptake of S-phase meiocytes revealed that TE5A mutants could succeed in completing DNA replication (Fig. 6A–C) compared to the wild-type (Fig. 6D–F).

View Article: PubMed Central - PubMed

ABSTRACT

Genic male sterility (GMS) has already been extensively utilized for hybrid rapeseed production. TE5A is a novel thermo-sensitive dominant GMS line in Brassica napus, however, its mechanisms of GMS remain largely unclear. Histological and Transmission electron microscopy (TEM) analyses of anthers showed that the male gamete development of TE5A was arrested at meiosis prophase I. EdU uptake of S-phase meiocytes revealed that the TE5A mutant could accomplish DNA replication, however, chromosomal and fluorescence in situ hybridization (FISH) analyses of TE5A showed that homologous chromosomes could not pair, synapse, condense and form bivalents. We then analyzed the transcriptome differences between young floral buds of sterile plants and its near-isogenic fertile plants through RNA-Seq. A total of 3,841 differentially expressed genes (DEGs) were obtained, some of which were associated with homologous chromosome behavior and cell cycle control during meiosis. Dynamic expression changes of selected candidate DEGs were then analyzed at different anther developmental stages. The present study not only demonstrated that the TE5A mutant had defects in meiotic prophase I via detailed cytological analysis, but also provided a global insight into GMS-associated DEGs and elucidated the mechanisms of GMS in TE5A through RNA-Seq.

No MeSH data available.