Project description:Maize LOB30 (Zm00001d036435) is a transcription factor and is specifically expressed in anthers. Our previous RNA-seq data showed that expression of some genes were upregualted in maize lob30 mutant maize anthers. To confirm these genes are the downstrem target genes, we generated proLOB30: GFP-LOB30 transgenic maize lines, collected stage 9 to stage10 anther materials and performed ChIP-seq using the GFP antibody.
Project description:In maize, 24-nt phased, secondary small interfering RNAs (phasiRNAs) are abundant in meiotic stage anthers, but their distribution and functions are not precisely known. Using laser capture microdissection we analyzed tapetal cells, meiocytes, and other somatic cells at several stages of anther development to establish the timing of 24-PHAS precursor transcripts and the 24-nt phasiRNA products. By integrating RNA and small RNA (sRNA) profiling plus single-molecule and sRNA FISH (smFISH or sRNA-FISH) spatial detection, we demonstrate that the tapetum is the primary site of 24-PHAS precursor and Dcl5 transcripts and the resulting 24-nt phasiRNAs. Interestingly, 24-nt phasiRNAs accumulate in all cell types, with the highest levels in meiocytes, followed by tapetum. Our data support the conclusion that 24-nt phasiRNAs are mobile from tapetum to meiocytes and to other somatic cells. We discuss possible roles for 24-nt phasiRNAs in anther cell types.
Project description:Agilent oligonucleotide arrays were used to profile gene expression in dissected maize anthers of 3 types of male-sterile plants and their fertile siblings at four stages of development: after anther initiation, at the rapid mitotic proliferation stage, pre-meiosis, and meiotic prophase I. The male-sterile mutants (ms23, msca, and mac1) lack a range of normal cell types resulting from a temporal progression of anther failure. By combining the data sets from the comparisons between individual sterile and fertile anthers, candidate genes predicted to play important roles during maize anther development were assigned to stages and to likely cell types. Keywords: anther development, maize, male sterility
Project description:Successful male gametogenesis involves orchestration of sequential gene regulation for somatic differentiation in pre-meiotic anthers. We report here the cloning of Male Sterile23 (Ms23), encoding an anther-specific predicted basic helix-loop-helix (bHLH) transcription factor required for tapetal differentiation; transcripts localize initially to the precursor secondary parietal cells then predominantly to daughter tapetal cells. In knockout ms23-ref mutant anthers, five instead of the normal four wall layers are observed. Microarray transcript profiling demonstrates a more severe developmental disruption in ms23-ref than in ms32 anthers, which possess a different bHLH defect. RNA-seq and proteomics data together with yeast two-hybrid assays suggest that MS23 along with MS32, bHLH122, and bHLH51 act sequentially as either homo- or heterodimers to choreograph tapetal development. Among them, MS23 is the earliest-acting factor, upstream of bHLH51 and bHLH122, controlling tapetal specification and maturation. In contrast, MS32 is constitutive and independently regulated and is required later than MS23 in tapetal differentiation. We characterized a maize (Zea mays) mutant line, ms23-ref, which the tapetal cells lack a dense cytoplasm and are not Binucleate -- the two characteristics of normal TP during meiosis. The meiocytes in ms23-ref fail to progress beyond meiotic prophase I. By profiling the small RNA population in ms23 mutants, we investigated the impact of ms23 mutation on small RNAs abundance in 0.4 mm, 0.7 mm, 1.0 mm, 1.5 mm and 2.0 mm anther. We also investigated the transcript abundance that were impacted in ms23 mutant by transcriptome profiling. The result provides an avenue for future research to understand the genetic networks and protein interactions among ms23, important for tapetal development in maize anther.
Project description:Global gene expression profiles of seven stages representing 29 days of anther development are analyzed using a 44K oligonucleotide array querying ~80% of maize protein-coding genes. Each anther stage expresses ~10,000 constitutive and ~10,000 or more transcripts restricted to one or a few stages. Keywords: anther development, maize
2008-12-01 | GSE12579 | GEO
Project description:RNA-Seq data of maize kernal mutant
| PRJNA957614 | ENA
Project description:Maize anther transcriptomes of ms33-6038 mutant
Project description:Transcriptomes from multiple pre-meiotic stages of wild type, mac1, and msca1 maize anthers were characterized by microarray hybridization. The goal was to characterize the developmental progression as the anther specifies five cell types and grows rapidly precedeing meiotic entry. The stages characterized were immature anther primordia (0.15 mm long in maize) containing just stem cells, through somatic and germinal cell fate specification (0.20 and 0.25 mm), mitotic proliferation (0.4 mm), and finally the birth of the middle layer and tapetum (0.7 mm). To obtain cell-type specific markers, at 0.7 mm we also compared whole anthers to collections of laser-microdissected anther cell types including the archesporial cells (pre-meiotic germinal cells), nutritive layers (middle layer and tapetum) and structural layers (endothecium and epidemis) of the anther lobe. keyword: anther development, maize, male-sterile