Cytoplasmic male sterility (CMS), encoded by the interacting mitochondrial and nuclear genes, causes pollen abortion or non-viability. CMS is widely used in agriculture and extensively studied in crops. Much less is known about CMS in wild species. We performed a comparative transcriptomic analysis of male sterile and fertile individuals of Silene vulgaris, a model plant for the study of gynodioecy, to reveal the genes responsible for pollen abortion in this species. We used RNA-seq datasets previously employed for the analysis of mitochondrial and plastid transcriptomes of female and hermaphrodite flower buds, making it possible to compare the transcriptomes derived from three genomes in the same RNA specimen. We assembled de novo transcriptomes for two haplotypes of S. vulgaris and identified differentially expressed genes between the females and hermaphrodites, associated with stress response or pollen development. The gene for alternative oxidase was downregulated in females. The genetic pathways controlling CMS in S. vulgaris are similar to those in crops. The high number of the differentially expressed nuclear genes contrasts with the uniformity of organellar transcriptomes across genders, which suggests these pathways are evolutionarily conserved and that selective mechanisms may shield organellar transcription against changes in the cytoplasmic transcriptome.
- Klíčová slova
- Silene vulgaris, cytoplasmic male sterility, differential gene expression,
- MeSH
- anotace sekvence MeSH
- buněčné jádro genetika MeSH
- down regulace genetika MeSH
- genová ontologie MeSH
- haplotypy genetika MeSH
- messenger RNA genetika metabolismus MeSH
- mitochondriální proteiny genetika metabolismus MeSH
- neplodnost rostlin genetika MeSH
- oxidační stres genetika MeSH
- oxidoreduktasy genetika metabolismus MeSH
- pyl genetika MeSH
- regulace genové exprese u rostlin * MeSH
- rostlinné proteiny genetika metabolismus MeSH
- Silene genetika fyziologie MeSH
- stanovení celkové genové exprese * MeSH
- transkriptom genetika MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- alternative oxidase MeSH Prohlížeč
- messenger RNA MeSH
- mitochondriální proteiny MeSH
- oxidoreduktasy MeSH
- rostlinné proteiny MeSH
The function of the plant hormone jasmonic acid (JA) in the development of tomato (Solanum lycopersicum) flowers was analyzed with a mutant defective in JA perception (jasmonate-insensitive1-1, jai1-1). In contrast with Arabidopsis (Arabidopsis thaliana) JA-insensitive plants, which are male sterile, the tomato jai1-1 mutant is female sterile, with major defects in female development. To identify putative JA-dependent regulatory components, we performed transcriptomics on ovules from flowers at three developmental stages from wild type and jai1-1 mutants. One of the strongly downregulated genes in jai1-1 encodes the MYB transcription factor SlMYB21. Its Arabidopsis ortholog plays a crucial role in JA-regulated stamen development. SlMYB21 was shown here to exhibit transcription factor activity in yeast, to interact with SlJAZ9 in yeast and in planta, and to complement Arabidopsis myb21-5 To analyze SlMYB21 function, we generated clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR associated protein 9 (Cas9) mutants and identified a mutant by Targeting Induced Local Lesions in Genomes (TILLING). These mutants showed female sterility, corroborating a function of MYB21 in tomato ovule development. Transcriptomics analysis of wild type, jai1-1, and myb21-2 carpels revealed processes that might be controlled by SlMYB21. The data suggest positive regulation of JA biosynthesis by SlMYB21, but negative regulation of auxin and gibberellins. The results demonstrate that SlMYB21 mediates at least partially the action of JA and might control the flower-to-fruit transition. .
- MeSH
- cyklopentany metabolismus MeSH
- down regulace MeSH
- fenotyp MeSH
- fertilita MeSH
- gibereliny metabolismus MeSH
- květy genetika fyziologie MeSH
- kyseliny indoloctové metabolismus MeSH
- mutace MeSH
- neplodnost rostlin MeSH
- ovoce genetika fyziologie MeSH
- oxylipiny metabolismus MeSH
- proteiny huseníčku genetika metabolismus MeSH
- regulace genové exprese u rostlin * MeSH
- regulátory růstu rostlin metabolismus MeSH
- rostlinné proteiny genetika metabolismus MeSH
- Solanum lycopersicum genetika fyziologie MeSH
- transkripční faktory genetika metabolismus MeSH
- vajíčko rostlin genetika fyziologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- cyklopentany MeSH
- gibereliny MeSH
- jasmonic acid MeSH Prohlížeč
- kyseliny indoloctové MeSH
- MYB21 protein, Arabidopsis MeSH Prohlížeč
- oxylipiny MeSH
- proteiny huseníčku MeSH
- regulátory růstu rostlin MeSH
- rostlinné proteiny MeSH
- transkripční faktory MeSH
BACKGROUND: Silene vulgaris (bladder campion) is a gynodioecious species existing as two genders - male-sterile females and hermaphrodites. Cytoplasmic male sterility (CMS) is generally encoded by mitochondrial genes, which interact with nuclear fertility restorer genes. Mitochondrial genomes of this species vary in DNA sequence, gene order and gene content. Multiple CMS genes are expected to exist in S. vulgaris, but little is known about their molecular identity. RESULTS: We assembled the complete mitochondrial genome from the haplotype KRA of S. vulgaris. It consists of five chromosomes, two of which recombine with each other. Two small non-recombining chromosomes exist in linear, supercoiled and relaxed circle forms. We compared the mitochondrial transcriptomes from females and hermaphrodites and confirmed the differentially expressed chimeric gene bobt as the strongest CMS candidate gene in S. vulgaris KRA. The chimeric gene bobt is co-transcribed with the Cytochrome b (cob) gene in some genomic configurations. The co-transcription of a CMS factor with an essential gene may constrain transcription inhibition as a mechanism for fertility restoration because of the need to maintain appropriate production of the necessary protein. Homologous recombination places the gene cob outside the control of bobt, which allows for the suppression of the CMS gene by the fertility restorer genes. We found the loss of three editing sites in the KRA mitochondrial genome and identified four sites with highly distinct editing rates between KRA and another S. vulgaris haplotypes (KOV). Three of these highly differentially edited sites were located in the transport membrane protein B (mttB) gene. They resulted in differences in MttB protein sequences between haplotypes. CONCLUSIONS: Frequent homologous recombination events that are widespread in plant mitochondrial genomes may change chromosomal configurations and also the control of gene transcription including CMS gene expression. Posttranscriptional processes, e.g. RNA editing shall be evaluated in evolutionary and co-evolutionary studies of mitochondrial genes, because they may change protein composition despite the sequence identity of the respective genes. The investigation of natural populations of wild species such as S. vulgaris are necessary to reveal important aspects of CMS missed in domesticated crops, the traditional focus of the CMS studies.
- MeSH
- cytochromy b genetika metabolismus MeSH
- editace RNA MeSH
- genom mitochondriální * MeSH
- haplotypy MeSH
- homologní rekombinace * MeSH
- membránové glykoproteiny genetika MeSH
- mitochondriální protonové ATPasy genetika MeSH
- mitochondrie genetika MeSH
- neplodnost rostlin genetika MeSH
- otevřené čtecí rámce genetika MeSH
- rostlinné proteiny genetika MeSH
- Silene genetika MeSH
- transkriptom MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- cytochromy b MeSH
- membránové glykoproteiny MeSH
- mitochondriální protonové ATPasy MeSH
- rostlinné proteiny MeSH
Chromatin is assembled by histone chaperones such as chromatin assembly factor CAF-1. We had noticed that vigor of Arabidopsis thaliana CAF-1 mutants decreased over several generations. Because changes in mutant phenotype severity over generations are unusual, we asked how repeated selfing of Arabidopsis CAF-1 mutants affects phenotype severity. CAF-1 mutant plants of various generations were grown, and developmental phenotypes, transcriptomes and DNA cytosine-methylation profiles were compared quantitatively. Shoot- and root-related growth phenotypes were progressively more affected in successive generations of CAF-1 mutants. Early and late generations of the fasciata (fas)2-4 CAF-1 mutant displayed only limited changes in gene expression, of which increasing upregulation of plant defense-related genes reflects the transgenerational phenotype aggravation. Likewise, global DNA methylation in the sequence context CHG but not CG or CHH (where H = A, T or C) changed over generations in fas2-4. Crossing early and late generation fas2-4 plants established that the maternal contribution to the phenotype severity exceeds the paternal contribution. Together, epigenetic rather than genetic mechanisms underlie the progressive developmental phenotype aggravation in the Arabidopsis CAF-1 mutants and preferred maternal transmission reveals a more efficient reprogramming of epigenetic information in the male than the female germline.
- Klíčová slova
- Arabidopsis thaliana, CAF-1, Chromatin, DNA methylation, Development, epigenetics, histone,
- MeSH
- alely MeSH
- Arabidopsis genetika MeSH
- epigeneze genetická * MeSH
- fenotyp MeSH
- fyziologický stres genetika MeSH
- genová ontologie MeSH
- metylace DNA genetika MeSH
- mutace genetika MeSH
- neplodnost rostlin MeSH
- proteiny huseníčku genetika metabolismus MeSH
- regulace genové exprese u rostlin MeSH
- sekvence nukleotidů MeSH
- semena rostlinná embryologie MeSH
- sestřihové faktory genetika metabolismus MeSH
- transkriptom genetika MeSH
- typy dědičnosti genetika MeSH
- vajíčko rostlin embryologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- At2g20020 protein, Arabidopsis MeSH Prohlížeč
- FAS protein, Arabidopsis MeSH Prohlížeč
- proteiny huseníčku MeSH
- sestřihové faktory MeSH
The interactions between mitochondria and nucleus substantially influence plant development, stress response and morphological features. The prominent example of a mitochondrial-nuclear interaction is cytoplasmic male sterility (CMS), when plants produce aborted anthers or inviable pollen. The genes responsible for CMS are located in mitochondrial genome, but their expression is controlled by nuclear genes, called fertility restorers. Recent explosion of high-throughput sequencing methods enabled to study transcriptomic alterations in the level of non-coding RNAs under CMS biogenesis. We summarize current knowledge of the role of nucleus encoded regulatory non-coding RNAs (long non-coding RNA, microRNA as well as small interfering RNA) in CMS. We also focus on the emerging data of non-coding RNAs encoded by mitochondrial genome and their possible involvement in mitochondrial-nuclear interactions and CMS development.
- Klíčová slova
- cytoplasmic male sterility, gene expression, global transcriptome, non-coding RNA, pollen development,
- MeSH
- autoinkompatibilita krytosemenných rostlin genetika MeSH
- Magnoliopsida fyziologie MeSH
- mitochondriální geny MeSH
- nekódující RNA genetika metabolismus MeSH
- neplodnost rostlin genetika MeSH
- pyl genetika fyziologie MeSH
- rostlinné geny MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- Názvy látek
- nekódující RNA MeSH
Cytoplasmic male sterility (CMS) is a widespread phenomenon in flowering plants caused by mitochondrial (mt) genes. CMS genes typically encode novel proteins that interfere with mt functions and can be silenced by nuclear fertility-restorer genes. Although the molecular basis of CMS is well established in a number of crop systems, our understanding of it in natural populations is far more limited. To identify CMS genes in a gynodioecious plant, Silene vulgaris, we constructed mt transcriptomes and compared transcript levels and RNA editing patterns in floral bud tissue from female and hermaphrodite full siblings. The transcriptomes from female and hermaphrodite individuals were very similar overall with respect to variation in levels of transcript abundance across the genome, the extent of RNA editing, and the order in which RNA editing and intron splicing events occurred. We found only a single genomic region that was highly overexpressed and differentially edited in females relative to hermaphrodites. This region is not located near any other transcribed elements and lacks an open-reading frame (ORF) of even moderate size. To our knowledge, this transcript would represent the first non-coding mt RNA associated with CMS in plants and is, therefore, an important target for future functional validation studies.
- Klíčová slova
- Cytoplasmic male sterility, Silene vulgaris, editing, mitochondrion, non-coding RNA, splicing, transcriptome.,
- MeSH
- editace RNA MeSH
- květy genetika růst a vývoj MeSH
- mitochondriální geny * MeSH
- nekódující RNA * MeSH
- neplodnost rostlin * MeSH
- rostlinné proteiny genetika metabolismus MeSH
- Silene genetika fyziologie MeSH
- transkriptom * MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
- Názvy látek
- nekódující RNA * MeSH
- rostlinné proteiny MeSH
The 7B-1 tomato (Solanum lycopersicum L. cv Rutgers) is a male-sterile mutant with enhanced tolerance to abiotic stress, which makes it a potential candidate for hybrid seed breeding and stress engineering. To underline the molecular mechanism regulating the male-sterility in 7B-1, transcriptomic profiles of the 7B-1 male-sterile and wild type (WT) anthers were studied using mRNA sequencing (RNA-Seq). In total, 768 differentially expressed genes (DEGs) were identified, including 132 up-regulated and 636 down-regulated transcripts. Gene ontology (GO) enrichment analysis of DEGs suggested a general impact of the 7B-1 mutation on metabolic processes, such as proteolysis and carbohydrate catabolic process. Sixteen candidates with key roles in regulation of anther development were subjected to further analysis using qRT-PCR and in situ hybridization. Cytological studies showed several defects associated with anther development in the 7B-1 mutant, including unsynchronized anther maturation, dysfunctional meiosis, arrested microspores, defect in callose degradation and abnormal tapetum development. TUNEL assay showed a defect in programmed cell death (PCD) of tapetal cells in 7B-1 anthers. The present study provides insights into the transcriptome of the 7B-1 mutant. We identified several genes with altered expression level in 7B-1 (including beta-1,3 glucanase, GA2oxs, cystatin, cysteine protease, pectinesterase, TA29, and actin) that could potentially regulate anther developmental processes, such as meiosis, tapetum development, and cell-wall formation/degradation.
- MeSH
- genetická transkripce * MeSH
- genová ontologie MeSH
- glukany metabolismus MeSH
- mutace * MeSH
- neplodnost rostlin genetika MeSH
- regulace genové exprese u rostlin * MeSH
- Solanum lycopersicum genetika metabolismus MeSH
- stanovení celkové genové exprese MeSH
- výpočetní biologie metody MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- callose MeSH Prohlížeč
- glukany MeSH
Sex determination in Rumex acetosa, a dioecious plant with a complex XY1 Y2 sex chromosome system (females are XX and males are XY1 Y2 ), is not controlled by an active Y chromosome but depends on the ratio between the number of X chromosomes and autosomes. To gain insight into the molecular mechanisms of sex determination, we generated a subtracted cDNA library enriched in genes specifically or predominantly expressed in female floral buds in early stages of development, when sex determination mechanisms come into play. In the present paper, we report the molecular and functional characterization of FEM32, a gene encoding a protein that shares a common architecture with proteins in different plants, animals, bacteria and fungi of the aerolysin superfamily; many of these function as β pore-forming toxins. The expression analysis, assessed by northern blot, RT-PCR and in situ hybridization, demonstrates that this gene is specifically expressed in flowers in both early and late stages of development, although its transcripts accumulate much more in female flowers than in male flowers. The ectopic expression of FEM32 under both the constitutive promoter 35S and the flower-specific promoter AP3 in transgenic tobacco showed no obvious alteration in vegetative development but was able to alter floral organ growth and pollen fertility. The 35S::FEM32 and AP3::FEM32 transgenic lines showed a reduction in stamen development and pollen viability, as well as a diminution in fruit set, fruit development and seed production. Compared with other floral organs, pistil development was, however, enhanced in plants overexpressing FEM32. According to these effects, it is likely that FEM32 functions in Rumex by arresting stamen and pollen development during female flower development. The aerolysin-like pore-forming proteins of eukaryotes are mainly involved in defence mechanisms against bacteria, fungi and insects and are also involved in apoptosis and programmed cell death (PCD), a mechanism that could explain the role of FEM32 in Rumex sex determination.
- Klíčová slova
- Rumex acetosa, FEM32 gene, aerolysin-like protein, male sterility, reproductive development, sex determination,
- MeSH
- bakteriální toxiny klasifikace genetika MeSH
- cytotoxické proteiny tvořící póry klasifikace genetika MeSH
- fylogeneze MeSH
- geneticky modifikované rostliny MeSH
- květy genetika růst a vývoj MeSH
- neplodnost rostlin genetika MeSH
- ovoce genetika růst a vývoj MeSH
- pyl genetika růst a vývoj MeSH
- regulace genové exprese u rostlin MeSH
- rostlinné proteiny klasifikace genetika MeSH
- Rumex genetika růst a vývoj MeSH
- sekvence aminokyselin MeSH
- sekvenční homologie aminokyselin MeSH
- semena rostlinná genetika růst a vývoj MeSH
- stanovení celkové genové exprese metody MeSH
- tabák genetika růst a vývoj MeSH
- vývojová regulace genové exprese MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- aerolysin MeSH Prohlížeč
- bakteriální toxiny MeSH
- cytotoxické proteiny tvořící póry MeSH
- rostlinné proteiny MeSH
Hybrid seed production in rice relies on cytoplasmic male sterility (CMS) induced by specific mitochondrial proteins, whose deleterious effects are suppressed by nuclear Restorer of Fertility (RF) genes. The majority of RF proteins belong to a specific clade of the RNA-binding pentatricopeptide repeat protein family. We have characterised 'restorer-of-fertility-like' (RFL) sequences from 13 Oryza genomes and the Brachypodium distachyon genome. The majority of the RFL sequences are found in genomic clusters located at two or three chromosomal loci with only a minor proportion being present as isolated genes. The RFL genomic cluster located on Oryza chromosome 10, the location of almost all known active rice RF genes, shows extreme variation in structure and gene content between species. We show evidence for homologous recombination events as an efficient mechanism for generating the huge repertoire of RNA sequence recognition motifs within RFL proteins and a major driver of RFL sequence evolution. The RFL sequences identified here will improve our understanding of the molecular basis of CMS and fertility restoration in plants and will accelerate the development of new breeding strategies.
- MeSH
- Brachypodium genetika MeSH
- chromozomy rostlin genetika MeSH
- genom rostlinný genetika MeSH
- neplodnost rostlin genetika MeSH
- rostlinné geny genetika MeSH
- rostlinné proteiny genetika MeSH
- rýže (rod) genetika MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- rostlinné proteiny MeSH
Ergot alkaloids are widely used in the pharmaceutical industry in drug preparations for treating migraines and Parkinson's disease, inducing uterine contraction, and other purposes. Phytopathogenic fungi of the genus Claviceps (e.g. C. purpurea) comprise a major biological source of ergot alkaloids. Worldwide industrial production of these alkaloids derives almost equally from two biotechnological procedures: submerged culture of the fungus in fermenters and field parasitic production in dormant fungal organs known as sclerotia (also termed ergot). Ergot yields from field cultivation are greatly affected by weather and also can be much reduced by pollen contamination from imperfectly male-sterile rye, as only unfertilized ovaries can be infected by C. purpurea spores. Two substances with gametocidal effect - maleic hydrazide and 2-chloroethylphosphonic acid - were tested during three consecutive seasons in small field experiments for the ability to induce or amplify the male sterility of rye as well as the impacts on germination of C. purpurea spores and general vitality of rye host plants. Maleic hydrazide was proven to be a highly effective gametocide on both a fertile rye variety and a variety with imperfectly induced cytoplasmic male sterility. It showed negligible effect on germination of C. purpurea spores. Both accurate dosaging of the active gametocidal compound and timing of the application just 2-3 weeks before onset of anthesis proved crucial to achieving high ergot yield with minimum grain impurities.
- MeSH
- hydrazid kyseliny maleinové aplikace a dávkování MeSH
- námelové alkaloidy biosyntéza izolace a purifikace MeSH
- neplodnost rostlin účinky léků fyziologie MeSH
- organofosforové sloučeniny aplikace a dávkování MeSH
- regulátory růstu rostlin farmakologie MeSH
- upregulace účinky léků fyziologie MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- zárodečné buňky rostlin účinky léků MeSH
- žito účinky léků metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- ethephon MeSH Prohlížeč
- hydrazid kyseliny maleinové MeSH
- námelové alkaloidy MeSH
- organofosforové sloučeniny MeSH
- regulátory růstu rostlin MeSH