Project description:In this study, we applied a quantitative proteomic approach using data-independent acquisition mass spectrometry (DIA-MS) to analyze protein expression profiles during three critical stages of anther development—pollen mother cell (Pms), tetrad (Tds), and mononuclear (Ms) stages—in the CMS line C2P5A and its maintainer line C2P5B in cotton. A total of 498 significantly differentially expressed proteins (DEPs) were identified, with 194 upregulated and 304 downregulated.
Project description:Cytoplasmic male sterility (CMS), a typically maternally inherited trait, causes a failure in producing functional pollen. Although the radish CMS has be widely used to produce hybrid varieties in breeding program, the molecular mechanism of CMS in radish is poorly understood. In this study, two radish CMS lines (HYBP-A and YH-A) and their corresponding maintainer lines (HYBP-B and YH-B) were used to identify genes potentially involving in CMS using Illumina pair-end sequencing. A total of 167.86 million clean sequence reads were generated from the eight libraries (two replicates for each line). These reads were eventually assembled into 130,240 unigenes. Of them, 67,173 (51.6%) unigenes were annotated for their function. Comparison of gene expression levels between CMS line and maintainer line revealed 5,893 differentially expressed genes (DEGs) in HYBP, and 3,739 DEGs inYH. There were 990 DEGs commonly identified in both HYBP and YH, with same direction of expression change in two CMS lines relative to their corresponding maintainer lines, which suggested these 990 DEGs is likely related to CMS of radish. The expression levels of 20 DEGs were further confirmed by real-time quantitative PCR (qRT-PCR). Two pathways and eight functional categories exhibited a significant enrichment with DEGs in HYBP, and one pathway and six functional categories were markedly enriched by DEGs in YH. Among these pathways/functional categories, four of them were enriched in both varieties. A series of candidate genes and pathways that may contribute to the CMS will be helpful for increasing our understanding for this trait in radish.
Project description:We employed a microarray-based transcriptomic status comparison of rice cytoplasmic male sterile (CMS) lines in order to categorize the nuclear gene expressions upon cytoplasmic substitution. In anthers at the uninucleate and bicellular pollen stages, we found that 8,199 genes significantly changed their expression in at least one of the CMS lines. We categorized the genes into 100 clusters by k-means clustering, and common obvious expression patterns were observed in W11, LD and BT.
Project description:We used a high-throughput proteomics method called label-free to compare protein abundance across a pepper CMS line and its isogenic maintainer line.This study explained the mechanisms of cytoplasmic male sterility and contribute to the improvement of pepper hybrid breeding.
Project description:Background: The use of cytoplasmic male sterility (CMS) in F1 hybrid seed production of chili pepper is increasingly popular. However, the molecular mechanisms of cytoplasmic male sterility and fertility restoration remain poorly understood due to limited transcriptomic and genomic data. Therefore, we analyzed the difference between a CMS line 121A and its near-isogenic restorer line 121C in transcriptome level using next generation sequencing technology (NGS), aiming to find out critical genes and pathways associated with the male sterility. Results: We generated approximately 53 million sequencing reads and assembled de novo, yielding 85,144 high quality unigenes with an average length of 643 bp. Among these unigenes, 27,191 were identified as putative homologs of annotated sequences in the public protein databases, 4,326 and 7,061 unigenes were found to be highly abundant in lines 121A and 121C, respectively. Many of the differentially expressed unigenes represent a set of potential candidate genes associated with the formation or abortion of pollen. Conclusions: Our study profiled anther transcriptomes of a chili pepper CMS line and its restorer line. The results shed the lights on the occurrence and recovery of the disturbances in nuclear-mitochondrial interaction and provide clues for further investigations. Anther transcriptomes of a chili pepper CMS line 121A and its nearisogenic restorer line 121C were generated by deep sequencing, using Illumina HiSeq 2000.
Project description:Here, we employed high-throughput sequencing to identify microRNAs in CMS and its maintainer fertile (MF) lines of Brassica juncea. We identified 197 known and 78 new candidate microRNAs during reproductive development of B. juncea. A total of 47 differentially expressed microRNAs between CMS and its maintainer fertile lines were discovered, according to their sequencing read number. Two samples from floral buds of CMS and MF lines.
Project description:Background: The use of cytoplasmic male sterility (CMS) in F1 hybrid seed production of chili pepper is increasingly popular. However, the molecular mechanisms of cytoplasmic male sterility and fertility restoration remain poorly understood due to limited transcriptomic and genomic data. Therefore, we analyzed the difference between a CMS line 121A and its near-isogenic restorer line 121C in transcriptome level using next generation sequencing technology (NGS), aiming to find out critical genes and pathways associated with the male sterility. Results: We generated approximately 53 million sequencing reads and assembled de novo, yielding 85,144 high quality unigenes with an average length of 643 bp. Among these unigenes, 27,191 were identified as putative homologs of annotated sequences in the public protein databases, 4,326 and 7,061 unigenes were found to be highly abundant in lines 121A and 121C, respectively. Many of the differentially expressed unigenes represent a set of potential candidate genes associated with the formation or abortion of pollen. Conclusions: Our study profiled anther transcriptomes of a chili pepper CMS line and its restorer line. The results shed the lights on the occurrence and recovery of the disturbances in nuclear-mitochondrial interaction and provide clues for further investigations.
Project description:ra05-11_cmsrapeseed - cms4:19 and at chr3 - -To understand how/if fertility and the mitochondrial background are interlinked. -To understand how the mitochondrial background influences the nuclear gene expression. -Compare and describe the total nuclear gene expression of CMS vs. fertile and CMS vs. Restored. -Describe and analyse genes that differ in expression (of special interest are genes that differs in both comparisons). -Group genes e.g. floral genes, highly expressed genes, transcription factors, nuclear encoded genes targeted for the mitochondrion. -Comparing two CMS-systems to elucidate differences and similarities between them. - Flower buds from four different B. napus lines (fertile cv Hanna, CMS-line 4:19, restored line 46, cv Hanna with the additon of A. thaliana chr 3) at two developmental stages (stages 0-5 and stage 8). Keywords: wt vs mutant comparison 16 dye-swap - CATMA arrays
Project description:High-throughput sRNA and degradome sequencing was applied in G1+HBP and its fertile type HBP to identify miRNAs and their targets during reproductive development. A total of 184 known miRNAs, 22 novel miRNAs and 86 target genes were identified. Some of the targets are transcription factors involved in floral development, such as auxin response factors (ARFs), SQUAMOSA promoter binding protein box (SBP-box), MYB, basic region-leucine zipper (bZIP), APETALA2 (AP2), transport inhibitor response 1 (TIR1), etc. Eight target genes were confirmed to be sliced by corresponding miRNAs using 5’RACE technology. Based on the sequencing abundance, 42 differentially expressed miRNAs between CMS line G1+HBP and fertile line HBP were discovered. Differential expression of miRNAs and their target genes was validated by quantitative RT-PCR and reciprocal expression patterns between some miRNAs and their targets were demonstrated. The regulation mechanism of miR167a was elucidated by yeast one-hybrid and dual-luciferase assay that a dehydrate responsive element binding (DREB) transcription factor bind miR167a promoter and transcriptionally repress miR167 expression. Our study revealed altered expression of miRNAs and their target genes in cytoplasmic male sterile pummelo (CMS) line and highlighted that miRNA regulatory network may be involved in nucleus-cytoplasmic cross talk of citrus CMS.
Project description:ra05-11_cmsrapeseed - cms4:19 and at chr3 - -To understand how/if fertility and the mitochondrial background are interlinked. -To understand how the mitochondrial background influences the nuclear gene expression. -Compare and describe the total nuclear gene expression of CMS vs. fertile and CMS vs. Restored. -Describe and analyse genes that differ in expression (of special interest are genes that differs in both comparisons). -Group genes e.g. floral genes, highly expressed genes, transcription factors, nuclear encoded genes targeted for the mitochondrion. -Comparing two CMS-systems to elucidate differences and similarities between them. - Flower buds from four different B. napus lines (fertile cv Hanna, CMS-line 4:19, restored line 46, cv Hanna with the additon of A. thaliana chr 3) at two developmental stages (stages 0-5 and stage 8). Keywords: wt vs mutant comparison