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1.
Gene ; 747: 144698, 2020 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-32325091

RESUMO

Stamen development is an important developmental process controlled by multiple internal and external factors. Developmental abnormalities of stamens can disrupt the structure and function of anthers, and then result in male sterility. As well known, APETELA 3 (AP3) has a clear function in regulating stamen development, which may impact in male sterility. However, the mechanisms of stamen development and male sterility controlled by AP3 are still not very clear, particular in Pak-choi (Brassica rapa ssp. chinensis). In this work, BcAP3 encoded a protein containing a MADS-box domain, which was a homolog of AtAP3, was identified in Pak-choi. Sequence alignments and phylogenetic analysis indicated that BcAP3 was highly similar to AtAP3. BcAP3 was shown to be localized to the nucleus and exhibited the potential of transcription factor. The transcript of BcAP3 was only expressed in flowers of Pak-choi, indicating that it may act in flower development. Overexpression of BcAP3 in Arabidopsis resulted in developmental abnormalities of anther wall and low vigor pollen, which were associated with the phenotype of male sterility. Expression levels of NST1 and NST2, involved in secondary wall thickening in anther walls, were significantly higher in the BcAP3-transgenic plants than in control plants, suggesting that BcAP3 may affect anther wall development by regulating NST1 and NST2. Taken together, our study demonstrated that BcAP3 could play an essential role in stamen development and male sterility.


Assuntos
Brassica rapa/crescimento & desenvolvimento , Brassica rapa/genética , Flores/crescimento & desenvolvimento , Flores/genética , Genes de Plantas , Proteínas de Domínio MADS/genética , Infertilidade das Plantas/genética , Proteínas de Plantas/genética , Sequência de Aminoácidos , Regulação da Expressão Gênica de Plantas , Proteínas de Domínio MADS/metabolismo , Fenótipo , Filogenia , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Pólen , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
2.
Mol Genet Genomics ; 295(3): 645-660, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32172356

RESUMO

Genetic male sterility (GMS) facilitates hybrid seed production in crops including cotton (Gossypium hirsutum). However, the genetic and molecular mechanisms specifically involved in this developmental process are poorly understood. In this study, small RNA sequencing, degradome sequencing, and transcriptome sequencing were performed to analyze miRNAs and their target genes during anther development in a GMS mutant ('Dong A') and its fertile wildtype (WT). A total of 80 known and 220 novel miRNAs were identified, 71 of which showed differential expressions during anther development. A further degradome sequencing revealed a total of 117 candidate target genes cleaved by 16 known and 36 novel miRNAs. Based on RNA-seq, 24, 11, and 21 predicted target genes showed expression correlations with the corresponding miRNAs at the meiosis, tetrad and uninucleate stages, respectively. In addition, a large number of differentially expressed genes were identified, most of which were involved in sucrose and starch metabolism, carbohydrate metabolism, and plant hormone signal transduction based on Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. The results of our study provide valuable information for further functional investigations of the important miRNAs and target genes involved in genetic male sterility and advance our understanding of miRNA regulatory functions during cotton anther development.


Assuntos
Regulação da Expressão Gênica no Desenvolvimento , Gossypium/crescimento & desenvolvimento , Gossypium/genética , MicroRNAs/genética , Mutação , Infertilidade das Plantas/genética , Proteínas de Plantas/genética , Flores/genética , Flores/crescimento & desenvolvimento , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Sequenciamento de Nucleotídeos em Larga Escala , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/crescimento & desenvolvimento , Transcriptoma
3.
Mol Biol Rep ; 47(2): 1275-1282, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31894465

RESUMO

The cytoplasmic male sterility (CMS) system is a useful tool for commercial hybrid cotton seed production. Two main CMS systems, CMS-D8 and CMS-D2, have been recognized with Rf2 and Rf1 as the restorer genes, respectively. The development of molecular markers tightly linked with restorer genes can facilitate the breeding of restorer lines. In this study, the InDel-1892 marker was developed to distinguish Rf2 and Rf1 simultaneously. Sequence alignment implied that CMS-D8-Rf2 has a 32 bp insertion and that CMS-D2-Rf1 has a 186 bp insertion at the InDel-1892 locus. The codominant marker was co-segregated with Rf1 and Rf2. Hence, this marker can be used for tracing Rf1 and Rf2 simultaneously and identifying the allele status at the restorer gene locus. The results of this study will facilitate efficient marker-assisted selection for restorer lines and hybrids of CMS systems.


Assuntos
Genes de Plantas , Marcadores Genéticos , Gossypium/genética , Mutação INDEL , Infertilidade das Plantas/genética , Mapeamento Cromossômico , Cruzamentos Genéticos , Loci Gênicos , Reprodutibilidade dos Testes , Análise de Sequência de DNA
4.
Genes (Basel) ; 11(1)2020 01 09.
Artigo em Inglês | MEDLINE | ID: mdl-31936663

RESUMO

Radish (Raphanus sativus L.) is an important root vegetable worldwide. The development of F1 hybrids, which are extensively used for commercial radish production, relies on cytoplasmic male sterility (CMS). To identify candidate genes responsible for CMS in NWB cytoplasm, we sequenced the normal and NWB CMS radish mitochondrial genomes via next-generation sequencing. A comparative analysis revealed 18 syntenic blocks and 11 unique regions in the NWB CMS mitogenome. A detailed examination indicated that orf463a was the most likely causal factor for male sterility in NWB cytoplasm. Interestingly, orf463a was identical to orf463, which is responsible for CMS in Dongbu cytoplasmic and genic male sterility (DCGMS) radish. Moreover, only structural variations were detected between the NWB CMS and DCGMS mitochondrial genomes, with no nucleotide polymorphisms (SNPs) or meaningful indels. Further analyses revealed these two mitochondrial genomes are coexisting isomeric forms belonging to the same mitotype. orf463a was more highly expressed in flower buds than in vegetative organs and its expression was differentially regulated in the presence of restorer of fertility (Rf) genes. orf463a was confirmed to originate from Raphanus raphanistrum. In this study, we identified a candidate gene responsible for the CMS in NWB cytoplasm and clarified the relationship between NWB CMS and DCGMS.


Assuntos
DNA Mitocondrial/genética , Infertilidade das Plantas/genética , Raphanus/genética , Sequência de Bases , Brassicaceae/genética , Mapeamento Cromossômico , Citoplasma , DNA Mitocondrial/metabolismo , Genoma Mitocondrial/genética , Sequenciamento de Nucleotídeos em Larga Escala , Mitocôndrias/genética
5.
BMC Plant Biol ; 20(1): 8, 2020 Jan 06.
Artigo em Inglês | MEDLINE | ID: mdl-31906856

RESUMO

BACKGROUND: Photoperiod and/or thermo-sensitive male sterility is an effective pollination control system in crop two-line hybrid breeding. We previously discovered the spontaneous mutation of a partially male sterile plant and developed a thermo-sensitive genic male sterile (TGMS) line 373S in Brassica napus L. The present study characterized this TGMS line through cytological observation, photoperiod/ temperature treatments, and genetic investigation. RESULTS: Microscopic observation revealed that the condensed cytoplasm and irregular exine of microspores and the abnormal degradation of tapetum are related to pollen abortion. Different temperature and photoperiod treatments in field and growth cabinet conditions indicated that the fertility alteration of 373S was mainly caused by temperature changes. The effects of photoperiod and interaction between temperature and photoperiod were insignificant. The critical temperature leading to fertility alteration ranged from 10 °C (15 °C/5 °C) to 12 °C (17 °C/7 °C), and the temperature-responding stage was coincident with anther development from pollen mother cell formation to meiosis stages. Genetic analysis indicated that the TGMS trait in 373S was controlled by one pair of genes, with male sterility as the recessive. Multiplex PCR analysis revealed that the cytoplasm of 373S is pol type. CONCLUSIONS: Our study suggested that the 373S line in B. napus has a novel thermo-sensitive gene Bnmst1 in Pol CMS cytoplasm background, and its fertility alteration is mainly caused by temperature changes. Our results will broaden the TGMS resources and lay the foundation for two-line hybrid breeding in B. napus.


Assuntos
Brassica napus/genética , Flores/citologia , Infertilidade das Plantas/genética , Brassica napus/anatomia & histologia , Flores/genética , Genes de Plantas , Fotoperíodo , Pólen/genética , Temperatura
6.
BMC Plant Biol ; 20(1): 10, 2020 Jan 07.
Artigo em Inglês | MEDLINE | ID: mdl-31910796

RESUMO

BACKGROUND: Cytoplasmic male sterility (CMS) plays a crucial role in the utilization of heterosis and various types of CMS often have different abortion mechanisms. Therefore, it is important to understand the molecular mechanisms related to anther abortion in wheat, which remain unclear at present. RESULTS: In this study, five isonuclear alloplasmic male sterile lines (IAMSLs) and their maintainer were investigated. Cytological analysis indicated that the abortion type was identical in IAMSLs, typical and stainable abortion, and the key abortive period was in the binucleate stage. Most of the 1,281 core shared differentially expressed genes identified by transcriptome sequencing compared with the maintainer in the vital abortive stage were involved in the metabolism of sugars, oxidative phosphorylation, phenylpropane biosynthesis, and phosphatidylinositol signaling, and they were downregulated in the IAMSLs. Key candidate genes encoding chalcone--flavonone isomerase, pectinesterase, and UDP-glucose pyrophosphorylase were screened and identified. Moreover, further verification elucidated that due to the impact of downregulated genes in these pathways, the male sterile anthers were deficient in sugar and energy, with excessive accumulations of ROS, blocked sporopollenin synthesis, and abnormal tapetum degradation. CONCLUSIONS: Through comparative transcriptome analysis, an intriguing core transcriptome-mediated male-sterility network was proposed and constructed for wheat and inferred that the downregulation of genes in important pathways may ultimately stunt the formation of the pollen outer wall in IAMSLs. These findings provide insights for predicting the functions of the candidate genes, and the comprehensive analysis of our results was helpful for studying the abortive interaction mechanism in CMS wheat.


Assuntos
Regulação da Expressão Gênica de Plantas/genética , Redes Reguladoras de Genes , Infertilidade das Plantas/genética , Transcriptoma/genética , Triticum , Biopolímeros/metabolismo , Carotenoides/metabolismo , Flores/citologia , Flores/ultraestrutura , Perfilação da Expressão Gênica/métodos , Ontologia Genética/estatística & dados numéricos , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Infertilidade das Plantas/fisiologia , Proteínas de Plantas/genética , Pólen/citologia , Pólen/ultraestrutura , Espécies Reativas de Oxigênio/metabolismo , Açúcares/metabolismo , Triticum/citologia , Triticum/genética , Triticum/metabolismo
7.
Mol Genet Genomics ; 295(1): 233-249, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31673754

RESUMO

In Chinese cabbage, hybrid seed production is performed using male sterility lines, an important approach to heterosis utilization. In this study, a stably inherited male sterile mutant msm was obtained from the 'FT'-doubled haploid line of Chinese cabbage using isolated microspore culture combined with 60Co γ-ray mutagenesis. The genetic backgrounds of 'FT' and msm were highly consistent; however, compared with wild-type 'FT', msm exhibited completely degenerated stamens and no pollen phenotype. Other characters showed no significant differences. Cytological observations revealed that stamen abortion in msm begins during the tetrad period and that tapetum cells were abnormally expanded and highly vacuolated, leading to microspore abortion. Genetic analysis indicated that the msm mutant phenotype is controlled by a single recessive nuclear gene. Comparative transcriptome analysis of 'FT' and msm flower buds using RNA-Seq technology revealed 1653 differentially expressed genes, among which, a large number associated with male sterility were detected, including 64 pollen development- and pollen tube growth-related genes, 94 pollen wall development-related genes, 11 phytohormone-related genes, and 16 transcription factor-related genes. An overwhelming majority of these genes were down-regulated in msm compared with 'FT'. Furthermore, KEGG pathway analysis indicated that a variety of carbohydrate metabolic and lipid metabolic pathways were significantly enriched, which may be related to pollen abortion. The expression patterns of 24 male sterility-related genes were analyzed using qRT-PCR. In addition, 24,476 single-nucleotide polymorphisms and 413,073 insertion-deletion events were specifically detected in msm. These results will facilitate elucidation of the regulatory mechanisms underlying male sterility in Chinese cabbage.


Assuntos
Brassica/genética , Genes de Plantas/genética , Infertilidade das Plantas/genética , Flores/genética , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica de Plantas/genética , Genes Recessivos/genética , Reguladores de Crescimento de Planta/genética , Proteínas de Plantas/genética , Pólen/genética , RNA-Seq/métodos , Transcriptoma/genética , Sequenciamento Completo do Exoma/métodos
8.
Int J Mol Sci ; 20(24)2019 Dec 11.
Artigo em Inglês | MEDLINE | ID: mdl-31835796

RESUMO

Male sterility is a valuable trait for genetic research and production application of wheat (Triticum aestivum L.). NWMS1, a novel typical genic male sterility mutant, was obtained from Shengnong 1, mutagenized with ethyl methane sulfonate (EMS). Microstructure and ultrastructure observations of the anthers and microspores indicated that the pollen abortion of NWMS1 started at the early uninucleate microspore stage. Pollen grain collapse, plasmolysis, and absent starch grains were the three typical characteristics of the abnormal microspores. The anther transcriptomes of NWMS1 and its wild type Shengnong 1 were compared at the early anther development stage, pollen mother cell meiotic stage, and binucleate microspore stage. Several biological pathways clearly involved in abnormal anther development were identified, including protein processing in endoplasmic reticulum, starch and sucrose metabolism, lipid metabolism, and plant hormone signal transduction. There were 20 key genes involved in the abnormal anther development, screened out by weighted gene co-expression network analysis (WGCNA), including SKP1B, BIP5, KCS11, ADH3, BGLU6, and TIFY10B. The results indicated that the defect in starch and sucrose metabolism was the most important factor causing male sterility in NWMS1. Based on the experimental data, a primary molecular regulation model of abnormal anther and pollen developments in mutant NWMS1 was established. These results laid a solid foundation for further research on the molecular mechanism of wheat male sterility.


Assuntos
Genes de Plantas , Mutação/genética , Infertilidade das Plantas/genética , Pólen/genética , Triticum/genética , Apoptose/genética , Análise por Conglomerados , Bases de Dados Genéticas , Regulação da Expressão Gênica de Plantas , Biblioteca Gênica , Ontologia Genética , Redes Reguladoras de Genes , Pólen/ultraestrutura , Análise de Componente Principal , Transcriptoma/genética , Triticum/ultraestrutura
9.
Int J Mol Sci ; 20(24)2019 Dec 05.
Artigo em Inglês | MEDLINE | ID: mdl-31817342

RESUMO

Cytoplasmic male sterility (CMS) is an important agronomic feature and provides an effective tool for heterosis utilization of crops. This study reports the comparative transcriptomic sketches between a novel allohexaploid cotton progeny CMS line LD6A and its maintainer line LD6B using de novo transcriptome sequencing technology at the pollen abortion stage. A total of 128,901 Unigenes were identified, in which 2007 were upregulated and 11,864 were downregulated. The significantly differentially expressed genes (DEGs) in LD6A show a distant and diverse genetic nature due to their distant hybrid hexaploidy progeny. Further analysis revealed that most of the DEGs participated in the tricarboxylic acid (TCA) cycle, oxidative phosphorylation, histone acetyltransferase activity, sepal development, stigma development, cotyledon development and microsporogenesis. A highly differentially expressed toxic protein, Abrin, was identified in the CMS line LD6A, which can catalyze the inactivation of ribosomes and consequently lead to cell death through the mitochondrial pathway in human cells. Twelve DEGs were selected randomly to validate transcriptome data using quantitative reverse-transcribed PCR (qRT-PCR). This study will contribute to new ideas and foundations related to the molecular mechanism of CMS and the innovation of cotton germplasm resources.


Assuntos
Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Gossypium , Infertilidade das Plantas , Proteínas de Plantas , Proteômica , Transcriptoma , Gossypium/genética , Gossypium/metabolismo , Proteínas de Plantas/biossíntese , Proteínas de Plantas/genética
10.
Int J Mol Sci ; 20(23)2019 Dec 02.
Artigo em Inglês | MEDLINE | ID: mdl-31810186

RESUMO

DNA methylation is an important epigenetic modification involved in multiple biological processes. Altered methylation patterns have been reported to be associated with male sterility in some plants, but their role in cotton cytoplasmic male sterility (CMS) remains unclear. Here, integrated methylome and transcriptome analyses were conducted between the CMS-D2 line ZBA and its near-isogenic maintainer line ZB in upland cotton. More methylated cytosine sites (mCs) and higher methylation levels (MLs) were found among the three sequence contexts in ZB compared to ZBA. A total of 4568 differentially methylated regions (DMRs) and 2096 differentially methylated genes (DMGs) were identified. Among the differentially expressed genes (DEGs) associated with DMRs (DMEGs), 396 genes were upregulated and 281 genes were downregulated. A bioinformatics analysis of these DMEGs showed that hyper-DEGs were significantly enriched in the "oxidative phosphorylation" pathway. Further qRT-PCR validation indicated that these hypermethylated genes (encoding the subunits of mitochondrial electron transport chain (ETC) complexes I and V) were all significantly upregulated in ZB. Our biochemical data revealed a higher extent of H2O2 production but a lower level of adenosine triphosphate (ATP) synthesis in CMS-D2 line ZBA. On the basis of the above results, we propose that disrupted DNA methylation in ZBA may disrupt the homeostasis of reactive oxygen species (ROS) production and ATP synthesis in mitochondria, triggering a burst of ROS that is transferred to the nucleus to initiate programmed cell death (PCD) prematurely, ultimately leading to microspore abortion. This study illustrates the important role of DNA methylation in cotton CMS.


Assuntos
Epigenoma/genética , Gossypium/genética , Infertilidade das Plantas/genética , Transcriptoma/genética , Citoplasma/genética , Metilação de DNA/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas/genética , Ontologia Genética , Pólen/genética , Pólen/crescimento & desenvolvimento
11.
BMC Genomics ; 20(1): 837, 2019 Nov 11.
Artigo em Inglês | MEDLINE | ID: mdl-31711411

RESUMO

BACKGROUND: Cytoplasmic male sterility (CMS) and its restoration of fertility (Rf) system is an important mechanism to produce F1 hybrid seeds. Understanding the interaction that controls restoration at a molecular level will benefit plant breeders. The CMS is caused by the interaction between mitochondrial and nuclear genes, with the CMS phenotype failing to produce functional anthers, pollen, or male gametes. Thus, understanding the complex processes of anther and pollen development is a prerequisite for understanding the CMS system. Currently it is accepted that the Rf gene in the nucleus restores the fertility of CMS, however the Rf gene has not been cloned. In this study, CMS line 8A and the Rf line R1, as well as a sterile pool (SP) of accessions and a restorer pool (RP) of accessions analyzed the differentially expressed genes (DEGs) between CMS and its fertility restorer using the conjunction of RNA sequencing and bulk segregation analysis. RESULTS: A total of 2274 genes were up-regulated in R1 as compared to 8A, and 1490 genes were up-regulated in RP as compared to SP. There were 891 genes up-regulated in both restorer accessions, R1 and RP, as compared to both sterile accessions, 8A and SP. Through annotation and expression analysis of co-up-regulated expressed genes, eight genes related to fertility restoration were selected. These genes encode putative fructokinase, phosphatidylinositol 4-phosphate 5-kinase, pectate lyase, exopolygalacturonase, pectinesterase, cellulose synthase, fasciclin-like arabinogalactan protein and phosphoinositide phospholipase C. In addition, a phosphatidylinositol signaling system and an inositol phosphate metabolism related to the fertility restorer of CMS were ranked as the most likely pathway for affecting the restoration of fertility in pepper. CONCLUSIONS: Our study revealed that eight genes were related to the restoration of fertility, which provides new insight into understanding the molecular mechanism of fertility restoration of CMS in Capsicum.


Assuntos
Capsicum/genética , Flores/genética , Regulação da Expressão Gênica de Plantas , Capsicum/crescimento & desenvolvimento , Capsicum/metabolismo , Fertilidade/genética , Flores/crescimento & desenvolvimento , Flores/metabolismo , Perfilação da Expressão Gênica , Ontologia Genética , Redes e Vias Metabólicas/genética , Infertilidade das Plantas , Análise de Sequência de RNA , Transcriptoma
12.
Int J Mol Sci ; 20(21)2019 Nov 05.
Artigo em Inglês | MEDLINE | ID: mdl-31694312

RESUMO

Cytoplasmic male sterility (CMS) is a maternally inherited trait used for hybrid production in plants, a novel kenaf CMS line 722HA was derived from the thermo-sensitive male-sterile mutant 'HMS' by recurrent backcrossing with 722HB. The line 722HA has great potential for hybrid breeding in kenaf. However, the underlying molecular mechanism that controls pollen abortion in 722HA remains unclear, thus limiting the full utilization of this line. To understand the possible mechanism governing pollen abortion in 722HA, cytological, transcriptomic, and biochemical analyses were carried out to compare the CMS line 722HA and its maintainer line 722HB. Cytological observations of the microspore development revealed premature degradation of the tapetum at the mononuclear stage, which resulted in pollen dysfunction. The k-means clustering analysis of differentially expressed genes (DEGs) revealed that these genes are related to processes associated with the accumulation of reactive oxygen species (ROS), including electron transport chain, F1F0-ATPase proton transport, positive regulation of superoxide dismutase (SOD), hydrogen peroxide catabolic, and oxidation-reduction. Biochemical analysis indicated that ROS-scavenging capability was lower in 722HA than in 722HB, resulting in an accumulation of excess ROS, which is consistent with the transcriptome results. Taken together, these results demonstrate that excessive ROS accumulation may affect the normal development of microspores. Our study provides new insight into the molecular mechanism of pollen abortion in 722HA and will promote further studies of kenaf hybrids.


Assuntos
Regulação da Expressão Gênica de Plantas , Hibiscus/genética , Infertilidade das Plantas/genética , Pólen/genética , Transcriptoma , Citoplasma/genética , Citoplasma/ultraestrutura , Hibiscus/crescimento & desenvolvimento , Hibiscus/ultraestrutura , Melhoramento Vegetal , Pólen/crescimento & desenvolvimento , Pólen/ultraestrutura , Espécies Reativas de Oxigênio/metabolismo
13.
BMC Genomics ; 20(1): 813, 2019 Nov 06.
Artigo em Inglês | MEDLINE | ID: mdl-31694534

RESUMO

BACKGROUND: Nsa cytoplasmic male sterility (CMS) is a novel alloplasmic male sterility system derived from somatic hybridization between Brassica napus and Sinapis arvensis. Identification of the CMS-associated gene is a prerequisite for a better understanding of the origin and molecular mechanism of this CMS. With the development of genome sequencing technology, organelle genomes of Nsa CMS line and its maintainer line were sequenced by pyro-sequencing technology, and comparative analysis of the organelle genomes was carried out to characterize the organelle genome composition of Nsa CMS as well as to identify the candidate Nsa CMS-associated genes. RESULTS: Nsa CMS mitochondrial genome showed a higher collinearity with that of S. arvensis than B. napus, indicating that Nsa CMS mitochondrial genome was mainly derived from S. arvensis. However, mitochondrial genome recombination of parental lines was clearly detected. In contrast, the chloroplast genome of Nsa CMS was highly collinear with its B. napus parent, without any evidence of recombination of the two parental chloroplast genomes or integration from S. arvensis. There were 16 open reading frames (ORFs) specifically existed in Nsa CMS mitochondrial genome, which could not be identified in the maintainer line. Among them, three ORFs (orf224, orf309, orf346) possessing chimeric and transmembrane structure are most likely to be the candidate CMS genes. Sequences of all three candidate CMS genes in Nsa CMS line were found to be 100% identical with those from S. arvensis mitochondrial genome. Phylogenetic and homologous analysis showed that all the mitochondrial genes were highly conserved during evolution. CONCLUSIONS: Nsa CMS contains a recombined mitochondrial genome of its two parental species with the majority form S. arvensis. Three candidate Nsa CMS genes were identified and proven to be derived from S. arvensis other than recombination of its two parental species. Further functional study of the candidate genes will help to identify the gene responsible for the CMS and the underlying molecular mechanism.


Assuntos
Brassica napus/genética , Brassica napus/fisiologia , Citoplasma/genética , Genes de Plantas/genética , Genômica , Organelas/genética , Infertilidade das Plantas/genética , Brassica napus/citologia , Genoma de Cloroplastos/genética , Genoma Mitocondrial/genética , Fases de Leitura Aberta/genética
14.
Int J Mol Sci ; 20(22)2019 Nov 06.
Artigo em Inglês | MEDLINE | ID: mdl-31698756

RESUMO

The cytoplasmic male sterility (CMS)/restorer-of-fertility system is an important tool to exploit heterosis during commercially hybrid seed production. The importance of long noncoding RNAs (lncRNAs) in plant development is recognized, but few analyses of lncRNAs during anther development of three-line hybrid cotton (CMS-D2 line A, maintainer line B, restorer-of-fertility line R) have been reported. Here, we performed transcriptome sequencing during anther development in three-line hybrid cotton. A total of 80,695 lncRNAs were identified, in which 43,347 and 44,739 lncRNAs were differentially expressed in A-B and A-R comparisons, respectively. These lncRNAs represent functional candidates involved in CMS and fertility restoration. GO analysis indicated that cellular hormone metabolic processes and oxidation-reduction reaction processes might be involved in CMS, and cellular component morphogenesis and small molecular biosynthetic processes might participate in fertility restoration. Additionally, 63 lncRNAs were identified as putative precursors of 35 miRNAs, and quantitative reverse transcription polymerase chain reaction (qRT-PCR) showed a similar expression pattern to RNA-seq data. Furthermore, construction of lncRNA regulatory networks indicated that several miRNA-lncRNA-mRNA networks might be involved in CMS and fertility restoration. Our findings provide systematic identification of lncRNAs during anther development and lays a solid foundation for the regulatory mechanisms and utilization in hybrid cotton breeding.


Assuntos
Perfilação da Expressão Gênica , Gossypium/genética , Gossypium/fisiologia , Infertilidade das Plantas/genética , RNA Longo não Codificante/genética , Sequência de Bases , Fertilidade/genética , Flores/genética , Flores/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas , Redes Reguladoras de Genes , Genoma de Planta , Hibridização Genética , MicroRNAs/genética , MicroRNAs/metabolismo , RNA Longo não Codificante/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
15.
Int J Mol Sci ; 20(20)2019 Oct 16.
Artigo em Inglês | MEDLINE | ID: mdl-31623069

RESUMO

Cotton (Gossypium hirsutum L.) is one of the most important cash crops worldwide. Cytoplasmic male sterility (CMS) is an excellent breeding system for exploitation of heterosis, which has great potential to increase crop yields. To understand the molecular mechanism of CMS in cotton, we compared transcriptome, cytomorphological, physiological and bioinformatics data between the CMS line C2P5A and its maintainer line C2P5B. By using high-throughput sequencing technology, 178,166 transcripts were assembled and 2013 differentially expression genes (DEGs) were identified at three different stages of C2P5A anther development. In this study, we identified DEGs associated with reactive oxygen species (ROS), peroxisomes, aldehyde dehydrogenases (ALDH), cytochrome oxidase subunit VI, and cytochrome P450, and DEGs associated with tapetum development, Jojoba acyl-CoA reductase-related male sterility protein, basic helix-loop-helix (bHLH) and MYB transcription factors. The abnormal expression of one of these genes may be responsible for the CMS C2P5A line. In gene ontology (GO) annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment, DEGs were mainly related to carbohydrate metabolism, amino acid metabolism, transport and catabolism, and signal transduction. Carbohydrate metabolism provides energy for anther development, starch and sucrose metabolism, fatty acid biosynthesis and metabolism and ascorbate and aldarate metabolism. These results showed that numerous genes and multiple complex metabolic pathways regulate cotton anther development. Weighted correlation network analysis (WGCNA) indicated that three modules, 'turquoise,' 'blue,' and 'green,' were specific for the CMS C2P5A line. The 'turquoise' and 'blue' modules were mainly related to carbohydrate metabolism, amino acid metabolism, energy metabolism, peroxisomes, pyruvate metabolism as well as fatty acid degradation. The 'green' module was mainly related to energy metabolism, carbon metabolism, translation, and lipid metabolism. RNA-sequencing and WGCNA polymerization modules were screened for key genes and pathways related to CMS in cotton. This study presents a new perspective for further research into the metabolic pathways of pollen abortion in the CMS C2P5A line and also provides a theoretical basis for its breeding and production.


Assuntos
Genes de Plantas , Gossypium/genética , Infertilidade das Plantas/genética , Transcriptoma , Biologia Computacional , Citoplasma , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Ontologia Genética , Imuno-Histoquímica , Redes e Vias Metabólicas , Anotação de Sequência Molecular , Fenótipo
16.
Plant Reprod ; 32(4): 371-380, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31620875

RESUMO

Nuclear male sterility (NMS) is a potential characteristic in crop recurrent selection and hybrid breeding. Mapping of nuclear male-sterile genes is key to utilizing NMS. Previously, we discovered a spontaneous soybean (Glycine max [L.] Merr.) male-sterile female-fertile mutant NJS-13H, which was conferred by a single recessive gene, designated msNJ. In this study, the msNJ was mapped to Chromosome 10 (LG O), and narrowed down between two SSR (simple sequence repeats) markers, BARCSOYSSR_10_794 and BARCSOYSSR_10_819 using three heterozygote-derived segregating populations, i.e., (NJS-13H × NN1138-2)F2, (NJS-13H × N2899)F2 and (NJS-13H)SPAG (segregating populations in advanced generations). This region spans approximately 1.32 Mb, where 27 genes were annotated according to the soybean reference genome sequence (Wm82.a2.v1). Among them, four genes were recognized as candidate genes for msNJ. Comparing to the physical locations of all the known male-sterile loci, msNJ is demonstrated to be a new male-sterile locus. This result may help the utilization and cloning of the gene.


Assuntos
Mapeamento Cromossômico , Genes de Plantas , Infertilidade das Plantas/genética , Soja/genética , Genes Recessivos , Repetições de Microssatélites
17.
Planta ; 250(6): 2159-2171, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31628536

RESUMO

MAIN CONCLUSION: In the wheat TCMS line YS3038, the anther development is inhibited from late uninucleate stage to the binucleate stage. The disruption of energy metabolism pathways by aberrant transcriptional regulation causes the male sterility under low temperatures. The utilization of thermosensitive male sterile (TMS) lines provides a basis for two-line breeding. Previous work, including morphological and cytological observations, has shown that the development process of the TMS line YS3038 is inhibited from the late uninucleate stage to the binucleate stage. Transcriptomics studies could now help to elucidate the overall expression of related genes in a specific reproductive process, revealing the metabolic network and its regulatory mechanism of the reproductive process from the transcription level. Considering the fertility characteristics of YS3038, three important stages for transcriptome analysis were determined to be the early uninucleate, late uninucleate and binucleate stages. The number of differentially expressed genes (DEGs) was found to be highest in the binucleate stage, and most were related to energy metabolism. Quantitative PCR analysis of selected genes related to energy metabolism revealed that their expression patterns were consistent with the sequencing results. Analysis of the fertility mechanism of YS3038 showed that although the tapetum of anthers was degraded in advance of the tetrad stage, the development of microspores did not result in obvious abnormalities until the binucleate stage, because the genes involved in energy metabolism pathways, including starch and sucrose metabolism (SSM), glycolysis, the tricarboxylic acid (TCA) cycle, oxidative phosphorylation, and respiration electron transport chain are differentially expressed under sterile and fertile conditions. Therefore, the pollen in YS3038 was sterile.


Assuntos
Metabolismo Energético/fisiologia , Fertilidade/genética , Fertilidade/fisiologia , Regulação da Expressão Gênica de Plantas , Infertilidade das Plantas/genética , Infertilidade das Plantas/fisiologia , Triticum/genética , Triticum/fisiologia , Perfilação da Expressão Gênica
18.
Genes Genomics ; 41(12): 1475-1492, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31576519

RESUMO

BACKGROUND: WS24-3A is a newly bred non-heading Chinese cabbage genic male-sterile line, in which sterility is controlled by a recessive gene, designated as Bra2ms. WS24-3A has been used for hybrid breeding. OBJECTIVE: To reveal the underlying molecular mechanisms responsible for the sterility of WS24-3A. METHODS: Cytological observation of the process of sterile/fertile anther development was performed to determine the tissue and stage in which sterility occurs. Phenotyping and transcriptomic analyses were performed to identify differentially expressed genes (DEGs) between sterile and fertile flower buds at different stages. RESULTS: Cytological analysis revealed no tetrads at stage 7 or at later stages of anther development, and the degradation of callose was delayed. Abnormal meiocytes were surrounded by sustaining callose that degenerated gradually in WS24-3A. Comparative transcript profiling identified 3282 DEGs during three anther developmental stages, namely, pre-meiotic anther, meiotic anther, and anthers with single-celled pollen stage. The difference in DEG percentage between up-regulated and down-regulated at meiotic anther stage was obviously larger than at the other two stages; further, most DEGs are important for male meiosis, callose synthesis and dissolution, and tapetum development. Ten DEGs were found to be involved in anther and pollen development, which were analyzed by quantitative PCR. CONCLUSION: Bra2ms affected gene expression in meiocytes and associated with callose synthesis, degradation and tapetum development. Our results provide clues to elucidate the molecular mechanism of genic male sterility in non-heading Chinese cabbage.


Assuntos
Brassica rapa/genética , Infertilidade das Plantas/genética , Brassica rapa/anatomia & histologia , Brassica rapa/crescimento & desenvolvimento , Brassica rapa/metabolismo , Flores/anatomia & histologia , Flores/crescimento & desenvolvimento , Perfilação da Expressão Gênica , Genes Recessivos , Glucanos/biossíntese , Meiose/genética , Fenótipo , Reação em Cadeia da Polimerase em Tempo Real , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
19.
J Genet ; 982019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31544786

RESUMO

Cytoplasmic male sterility (CMS) is an important trait for large-scale hybrid seed production which avoids manual emasculation and undesired horizontal spread of pollen. Rearrangements in mitochondrial genome in terms of deletions and insertions are frequent causes leading to CMS. Mitochondrial ATP synthase is a multisubunit molecular machine which is involved in synthesis of ATP. In this study, three mutations in ATPase subunit 6 were identified and their cosegregation with male sterility was established using tobacco male sterile hybrids and Nicotiana suvaolensis. A breeder friendly Kompetitive allele specific polymerase chain reaction (KASP) SNP marker was developed for high throughput and quick genotyping. Introgression of this trait into selected germplasm lines (n = 9) was achieved based on foreground for CMS and background selection for recurrent parent using KASP marker and 50K custom tobacco SNP array, respectively. Analysis of genotyping data from SNP array revealed the presence of 88-99% of recurrent parent genome in BC3F1 plants. The selected BC3F1 plants exhibit CMS and are indistinguishable from the fertile recurrent parent (germplasm) in terms of plant morphology.


Assuntos
ATPases Mitocondriais Próton-Translocadoras/genética , Infertilidade das Plantas/genética , Pólen/genética , Tabaco/genética , Alelos , DNA Mitocondrial/genética , Marcadores Genéticos , Genótipo , Análise de Sequência com Séries de Oligonucleotídeos , Fenótipo , Reação em Cadeia da Polimerase , Polimorfismo de Nucleotídeo Único
20.
J Genet ; 982019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31544787

RESUMO

Cytoplasmic male sterility (CMS) is widely used for hybrid seed production in cultivated Solanaceae species. However, there is very limited information about CMS-Rf genetic systems in potato (Solanum tuberosum). Studying the CMS-Rf systems in potato is both of theoretical and practical significance due to the emergence of a new revolutionary strategy of reinventing potato as adiploid inbred line-based crop to develop F1 hybrid seed potato breeding (Lindhout et al. 2011; Jansky et al. 2016). To search for potato Rf gene candidates, the comparative genetic approach was applied. Based on similarity to petunia Rf-PPR592 gene, 38 fragments were identified in five loci of the whole-genome nucleotide sequence of the accession DM 1-3 516 R44 S. tuberosum Phureja group (https://blast.ncbi.nlm.nih.gov/Blast.cgi). The putative encoded mitochondrial proteins have 589-597 amino acid residues, similarto RF-PPR proteins of petunia and chili pepper and contain 14 or 15 PPR motifs. Primers have been developed flanking the most variable 782-865 bp regions of the selected loci, and polymorphism of the cloned fragments has been investigated in a subset of nine potato genotypes. The amplified fragments included seven or eight PPR motifs and lacked introns. The SNP frequencies ranged from 7.0 to 19.8% depending on the locus, while the ratio of nonsynonymous to synonymous substitutions varied between 0.9 and 2.1. Positions 1, 3 and 6 were the most variable in the studied PPR motifs. Our results demonstrated that the analysed sequences belong to the RFL-PPR gene subfamily and may be considered as Rf gene candidates in potato.


Assuntos
Proteínas de Plantas/genética , Polimorfismo Genético , Solanum tuberosum/genética , Motivos de Aminoácidos , Sequência de Aminoácidos , Fertilidade/genética , Íntrons , Filogenia , Infertilidade das Plantas , Proteínas de Plantas/química , Pólen/genética , Pólen/crescimento & desenvolvimento
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