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1.
Yi Chuan ; 41(8): 703-715, 2019 Aug 20.
Artigo em Chinês | MEDLINE | ID: mdl-31447421

RESUMO

Hybrid rice has contributed greatly to global food security. Cytoplasmic male sterility (CMS) and photo/ thermo sensitive genic male sterility (P/TGMS) are genetic bases for three-line and two-line hybrid rice production, respectively. In contrast, (sub-) specific hybrid sterility (HS) is a major barrier for utilization of hybrid vigor of distant hybrid rice. Therefore, understanding the molecular regulatory mechanism of rice fertility is a key technical issue for hybrid rice industry, and a long-standing basic scientific issue for nuclear-cytoplasmic interaction and reproductive isolation. Chinese geneticists of plant sciences have made tremendous contributions on the molecular genetic basis of rice fertility related to hybrid rice production. Here, we review the development of hybrid rice production systems in China and summarize current advance on genetic basis and molecular mechanism of CMS, P/TGMS, and HS involved in hybrid rice. We also discuss problems of hybrid rice production in China and point out new direction for future utilization of heterosis in rice.


Assuntos
Hibridização Genética , Oryza/genética , Oryza/fisiologia , Infertilidade das Plantas/genética , China , Fertilidade/genética , Vigor Híbrido
2.
BMC Plant Biol ; 19(1): 252, 2019 Jun 11.
Artigo em Inglês | MEDLINE | ID: mdl-31185903

RESUMO

BACKGROUND: Thermo-sensitive male-sterility based on Aegilops kotschyi cytoplasm (K-TCMS) plays an important role in hybrid wheat breeding. This has important possible applications in two-line hybrid wheat breeding but the genetic basis and molecular regulation mechanism related to fertility restoration are poorly understood. In this study, comparative transcriptome profiling based on RNA sequencing was conducted for two near-isogenic lines comprising KTM3315R and its sterile counterpart KTM3315A, a total of six samples (3 repetitions per group), in order to identify fertility restoration genes and their metabolic pathways. RESULTS: In total, 2642 significant differentially expressed genes (DEGs) were detected, among which 1238 were down-regulated and 1404 were up-regulated in fertile anthers. Functional annotation enrichment analysis identified important pathways related to fertility restoration, such as carbohydrate metabolism, phenylpropanoid metabolism and biosynthesis, as well as candidate genes encoding pectin methylesterase and flavanone 3-hydroxylase. Moreover, transcription factor analysis showed that a large number of DEGs were mainly involved with the WRKY, bHLH, and MYB transcription factor families. Determination of total soluble sugar and flavonoid contents demonstrated that important metabolic pathways and candidate genes are associated with fertility restoration. Twelve DEGs were selected and detected by quantitative reverse-transcribed PCR, and the results indicated that the transcriptome sequencing results were reliable. CONCLUSIONS: Our results indicate that identified DEGs were related to the fertility restoration and they proved to be crucial in Aegilops kotschyi cytoplasm. These findings also provide a basis for exploring the molecular regulation mechanism associated with wheat fertility restoration as well as screening and cloning related genes.


Assuntos
Aegilops/genética , Melhoramento Vegetal , Infertilidade das Plantas/genética , Proteínas de Plantas/genética , Transcriptoma , Triticum/genética , Citoplasma/fisiologia , Perfilação da Expressão Gênica , Redes e Vias Metabólicas/genética , Proteínas de Plantas/metabolismo , Triticum/fisiologia
3.
Plant Physiol Biochem ; 141: 231-239, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31195253

RESUMO

Anther/pollen development is a highly programmed process in flowering plants. However, the molecular mechanism of regulating anther/pollen development is still largely unclear so far. Here, we report a cotton WRKY transcription factor (GhWRKY22) that functions in anther/pollen development. Quantitative RT-PCR and GUS activity analyses revealed that GhWRKY22 is predominantly expressed in the late developing anther/pollen of cotton. The transgenic Arabidopsis plants expressing GhWRKY22 displayed the male fertility defect with the fewer viable pollen grains. Expression of the genes involved in jasmonate (JA) biosynthesis was up-regulated, whereas expression of the JA-repressors (JAZ1 and JAZ8) was down-regulated in the transgenic Arabidopsis plants expressing GhWRKY22, compared with those in wild type. Yeast one-hybrid and ChIP-qPCR assays demonstrated that GhWRKY22 modulated the expression of JAZ genes by directly binding to their promoters for regulating anther/pollen development. Yeast two-hybrid assay indicated that GhMYB24 could interact with GhJAZ8-A and GhJAZ13-A. Furthermore, expression of AtMYB24, AtPAL2 and AtANS2 was enhanced in the transgenic Arabidopsis plants, owing to GhWRKY22 overexpression. Taking the data together, our results suggest that GhWRKY22 acts as a transcriptional repressor to regulate anther/pollen development possibly by modulating the expression of the JAZ genes.


Assuntos
Gossypium/metabolismo , Pólen/fisiologia , Fatores de Transcrição/metabolismo , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Núcleo Celular/metabolismo , Ciclopentanos/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Hipocótilo/metabolismo , Oxilipinas/metabolismo , Fenótipo , Infertilidade das Plantas/genética , Proteínas de Plantas/metabolismo , Caules de Planta/metabolismo , Plantas Geneticamente Modificadas , Regiões Promotoras Genéticas , Sementes/metabolismo , Ativação Transcricional , Transgenes , Técnicas do Sistema de Duplo-Híbrido
4.
Nat Commun ; 10(1): 2501, 2019 06 07.
Artigo em Inglês | MEDLINE | ID: mdl-31175302

RESUMO

Hybrid sterility (HS) between Oryza sativa (Asian rice) and O. glaberrima (African rice) is mainly controlled by the S1 locus. However, our limited understanding of the HS mechanism hampers utilization of the strong interspecific heterosis. Here, we show that three closely linked genes (S1A4, S1TPR, and S1A6) in the African S1 allele (S1-g) constitute a killer-protector system that eliminates gametes carrying the Asian allele (S1-s). In Asian-African rice hybrids (S1-gS1-s), the S1TPR-S1A4-S1A6 interaction in sporophytic tissues generates an abortion signal to male and female gametes. However, S1TPR can rescue S1-g gametes, while the S1-s gametes selectively abort for lacking S1TPR. Knockout of any of the S1-g genes eliminates the HS. Evolutionary analysis suggests that S1 may have arisen from newly evolved genes, multi-step recombination, and nucleotide variations. Our findings will help to overcome the interspecific reproductive barrier and use Asian-African hybrids for increasing rice production.


Assuntos
Quimera/genética , Oryza/genética , Infertilidade das Plantas/genética , Proteínas de Plantas/genética , Alelos , Técnicas de Inativação de Genes , Células Germinativas Vegetais
5.
Plant Physiol Biochem ; 141: 73-82, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31132695

RESUMO

Polyploidy could increase the interactions of pollen sterility loci and Sb locus interaction cause higher pollen abortion than other loci. Therefore, we focused on the interaction at Sb pollen sterility locus in autotetraploid rice compared to diploid rice hybrid using the near-isogenic lines in the present study. Cytological observations indicated that interaction at Sb locus cause high pollen sterility (69.9%) and abnormal chromosome behavior (37.02%) at Metaphase II in autotetraploid rice hybrid. A total of 139 meiosis-related or meiosis stage-specific genes were detected in the autotetraploid rice hybrid harboring interaction at Sb locus and 27 of these meiosis-related or specific genes displayed significant down-regulation, including four pollen fertility related genes (Rad51, XRI1, PSS1 and MIL1). These results revealed a stronger interaction at Sb pollen sterility locus than other loci, which cause down-regulation of many important meiosis-related genes that were associated with higher pollen sterility in autotetraploid rice hybrids.


Assuntos
Cruzamentos Genéticos , Meiose/genética , Oryza/genética , Infertilidade das Plantas/genética , Pólen/genética , Alelos , Biologia Celular , Cromossomos de Plantas , Regulação para Baixo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Genótipo , Heterozigoto , Tetraploidia , Transcriptoma
6.
Plant Cell Physiol ; 60(7): 1604-1618, 2019 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-31076750

RESUMO

K-type cytoplasmic male sterility (KCMS) lines were ideal material for three-line hybrid wheat system due to the major role in hybrid wheat production. In this study, the morphology of developing microspore and mature pollen was compared between a KCMS line and its near-isogenic restorer line (KCMS-NIL). The most striking difference is that the microspore was unable to develop into tricellular pollen in the KCMS line. MicroRNA plays vital roles in flowering and gametophyte development. Small RNA sequencing identified a total of 274 known and 401 novel miRNAs differentially expressed between two lines or two developmental stages. Most of miRNAs with high abundance were differentially expressed at the uninucleate stage, and their expression level recovered or remained at the binucleate stage. Further degradome sequencing identified target genes which were mainly enriched in transcription regulation, phytohormone signaling and RNA degradation pathways. Combining with the transcriptome data, a correlation was found between the abnormal anther development, such as postmeiotic mitosis cessation, deformative pollen wall and the chromosome condensation of the vegetative cell, and the alterations in the related miRNA and their targets expression profiles. According to the correlation and pathway analysis, we propose a hypothetic miRNA-mediated network for the control of KCMS restoration.


Assuntos
MicroRNAs/genética , Infertilidade das Plantas/genética , Triticum/genética , Flores/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas/genética , Biblioteca Gênica , Ontologia Genética , Genes de Plantas/genética , Genes de Plantas/fisiologia , MicroRNAs/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Reprodução/genética , Análise de Sequência de RNA , Triticum/metabolismo
7.
Planta ; 250(2): 507-518, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31098709

RESUMO

MAIN CONCLUSION: 'Petaloid' cytoplasmic male sterility is commonly used as a stable genetic mechanism in carrot hybrid breeding. Its introgression in tropical carrot showed morphometric changes and molecular markers were identified for detection at early stage. Cytoplasmic male sterility (CMS) is the only genetic mechanism in carrot for commercial exploitation of heterosis and production of low cost affordable hybrid seeds. The 'petaloid' CMS system is stable and commonly used in hybrid breeding in temperate carrot but there is no information available on existence of natural CMS system in tropical Asiatic carrot. Therefore, the present study was aimed to investigate morphometric traits and organizational features of cytoplasmic atp9 gene sequences in newly converted CMS lines (BC4-7) of tropical carrot. The CMS lines had root traits at par with fertile counterparts while floral traits had variation. Petal colour and length, petaloids colour and shape and style length showed differences among the CMS lines and with their maintainers. Molecular markers are effective to establish male sterility at genetic level, for this, six fixed and stable CMS lines were screened with seven novel primer combinations. Out of which five pairs produced clearly distinguishable bands in CMS lines and their fertile counterparts. The study confirmed that the region between 3' end of atp9-1/atp9-3 gene and 5' end of region of homology to Arabidopsis thaliana mtDNA is ideal for developing the trait specific markers. These new CMS lines have potential to use in hybrid development and molecular markers will be useful to confirm male sterility to rogue out fertile plants.


Assuntos
Daucus carota/genética , Infertilidade das Plantas/genética , Proteínas de Plantas/genética , Arabidopsis/anatomia & histologia , Arabidopsis/genética , Arabidopsis/fisiologia , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Quimera , Citoplasma/genética , DNA Mitocondrial/genética , Daucus carota/anatomia & histologia , Daucus carota/fisiologia , Marcadores Genéticos/genética , ATPases Mitocondriais Próton-Translocadoras/genética , ATPases Mitocondriais Próton-Translocadoras/metabolismo , Melhoramento Vegetal , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas
8.
Theor Appl Genet ; 132(7): 2137-2154, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31016347

RESUMO

KEY MESSAGE: Genome-wide analysis of maize GPAT gene family, cytological characterization of ZmMs33/ZmGPAT6 gene encoding an ER-localized protein with four conserved motifs, and its molecular breeding application in maize. Glycerol-3-phosphate acyltransferase (GPAT) mediates the initial step of glycerolipid biosynthesis and plays pivotal roles in plant growth and development. Compared with GPAT genes in Arabidopsis, our understanding to maize GPAT gene family is very limited. Recently, ZmMs33 gene has been identified to encode a sn-2 GPAT protein and control maize male fertility in our laboratory (Xie et al. in Theor Appl Genet 131:1363-1378, 2018). However, the functional mechanism of ZmMs33 remains elusive. Here, we reported the genome-wide analysis of maize GPAT gene family and found that 20 maize GPAT genes (ZmGPAT1-20) could be classified into three distinct clades similar to those of ten GPAT genes in Arabidopsis. Expression analyses of these ZmGPAT genes in six tissues and in anther during six developmental stages suggested that some of ZmGPATs may play crucial roles in maize growth and anther development. Among them, ZmGPAT6 corresponds to the ZmMs33 gene. Systemic cytological observations indicated that loss function of ZmMs33/ZmGPAT6 led to defective anther cuticle, arrested degeneration of anther wall layers, abnormal formation of Ubisch bodies and exine and ultimately complete male sterility in maize. The endoplasmic reticulum-localized ZmMs33/ZmGPAT6 possessed four conserved amino acid motifs essential for acyltransferase activity, while ZmMs33/ZmGPAT6 locus and its surrounding genomic region have greatly diversified during evolution of gramineous species. Finally, a multi-control sterility system was developed to produce ms33 male-sterile lines by using a combination strategy of transgene and marker-assisted selection. This work will provide useful information for further deciphering functional mechanism of ZmGPAT genes and facilitate molecular breeding application of ZmMs33/ZmGPAT6 gene in maize.


Assuntos
Família Multigênica , Melhoramento Vegetal , Infertilidade das Plantas/genética , Zea mays/genética , Sequência de Aminoácidos , Flores/genética , Flores/fisiologia , Genes de Plantas , Estudos de Associação Genética , Microscopia Eletrônica de Varredura , Filogenia , Plantas Geneticamente Modificadas , Pólen/ultraestrutura , Sintenia , Zea mays/fisiologia
9.
Theor Appl Genet ; 132(7): 2125-2135, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31020387

RESUMO

KEY MESSAGE: Tomato male sterile-1526 locus was fine-mapped to an interval of 44.6 kb, and a B-class MADS-box gene TM6 was identified as the candidate gene. Male sterile lines have been widely used for hybrid seed production in many crop plants. The tomato male sterile-1526 (ms-1526) mutant displays abnormal stamens and exerted stigmas and is suitable for practical use. In this study, the ms-1526 locus was fine-mapped to a 44.6 kb interval that contained four putative genes. Thereinto, Solyc02g084630 encodes tomato B-class MADS-box gene TM6 (syn. TDR6), which plays an important role in stamen development. Sequencing revealed that there was a 12.7 kb deletion in the ms-1526 region, where the promoter and first four exons of the TM6 gene were absent. ms-1547, an allele of ms-1526, also contained the same deletion in the TM6 gene. And the other allele ms-15 mutant contained a single-nucleotide polymorphism (SNP, C to A) in the coding region of the TM6 gene, which led to a missense mutation (G to W). The codominant insertion/deletion (InDel) marker MS26D and codominant derived cleaved amplified polymorphic sequence (dCAPS) marker MS15C were developed based on the deletion and SNP, respectively. A real-time quantitative reverse-transcription PCR showed that expression of the TM6 gene was barely detectable in the flowers of the ms-1526 and ms-1547 mutants. In addition, other floral organ identity genes, pollen development marker genes, and pistil marker genes were differentially expressed between wild type and mutant flowers. These findings may facilitate functional analysis of the TM6 gene and help in the marker-assisted selection of ms-15 and its alleles in tomato breeding.


Assuntos
Flores/fisiologia , Lycopersicon esculentum/genética , Proteínas de Domínio MADS/genética , Infertilidade das Plantas/genética , Proteínas de Plantas/genética , Alelos , Mapeamento Cromossômico , Flores/genética , Marcadores Genéticos , Genótipo , Mutação INDEL , Lycopersicon esculentum/fisiologia , Fenótipo , Polimorfismo de Nucleotídeo Único , Deleção de Sequência
10.
Yi Chuan ; 41(3): 262-270, 2019 Mar 20.
Artigo em Chinês | MEDLINE | ID: mdl-30872262

RESUMO

It has always been a challenge to combine research progress with undergraduate laboratory teaching. Herein we designed a comprehensive experiment to compare classical Mendelian segregation and non-Mendelian distorted segregation by utilizing a rice material (DSSL) containing F1 hybrid male sterility locus S23 constructed previously in our research project. Using the four SSR markers located on two chromosomes of rice, the genotypes of the F2 population and the two parents were analyzed, and the phenotypes of the pollen fertility of the two parents and their F1 plants were investigated. The results not only verified segregation law at the molecular level, but also fully demonstrated the distorted segregation in both genotypes and phenotypes, thus deepening students' understandings of plant genetics and the relationship between genotypes and phenotypes, inspiring students' interests in genetics experiments, and enhancing students' consciousness and enthusiasm for experimental learning. On the basis of this, a sustainable development idea of transforming scientific research progress into teaching applications was conceived to promote the reform and innovation of genetics laboratory teaching.


Assuntos
Segregação de Cromossomos , Genética/educação , Aprendizagem , Oryza/genética , Marcadores Genéticos , Genótipo , Repetições de Microssatélites , Infertilidade das Plantas/genética , Pólen , Estudantes
11.
Int J Mol Sci ; 20(6)2019 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-30897816

RESUMO

Genic male sterility (GMS) mutant is a useful germplasm resource for both theory research and production practice. The identification and characterization of GMS genes, and assessment of male-sterility stability of GMS mutant under different genetic backgrounds in Zea may (maize) have (1) deepened our understanding of the molecular mechanisms controlling anther and pollen development, and (2) enabled the development and efficient use of many biotechnology-based male-sterility (BMS) systems for hybrid breeding. Here, we reported a complete GMS mutant (ms20), which displays abnormal anther cuticle and pollen development. Its fertility restorer gene ZmMs20 was found to be a new allele of IPE1 encoding a glucose methanol choline (GMC) oxidoreductase involved in lipid metabolism in anther. Phylogenetic and microsynteny analyses showed that ZmMs20 was conserved among gramineous species, which provide clues for creating GMS materials in other crops. Additionally, among the 17 maize cloned GMS genes, ZmMs20 was found to be similar to the expression patterns of Ms7, Ms26, Ms6021, APV1, and IG1 genes, which will give some clues for deciphering their functional relationships in regulating male fertility. Finally, two functional markers of ZmMs20/ms20 were developed and tested for creating maize ms20 male-sterility lines in 353 genetic backgrounds, and then an artificial maintainer line of ms20 GMS mutation was created by using ZmMs20 gene, ms20 mutant, and BMS system. This work will promote our understanding of functional mechanisms of male fertility and facilitate molecular breeding of ms20 male-sterility lines for hybrid seed production in maize.


Assuntos
Infertilidade das Plantas/fisiologia , Zea mays/genética , Filogenia , Melhoramento Vegetal , Infertilidade das Plantas/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Zea mays/classificação
12.
BMC Plant Biol ; 19(1): 109, 2019 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-30894127

RESUMO

BACKGROUND: Two-line hybrid rice with high yield potential is increasingly popular and the photo- and temperature-sensitive male sterile line is one of the basic components for two-line hybrid rice breeding. The development of male sterile lines through conventional breeding is a lengthy and laborious process, whereas developing thermo-sensitive genic male sterile (TGMS) lines for two-line hybrid breeding by editing a temperature-sensitivity gene by CRISPR/Cas9 is efficient and convenient. RESULTS: Here, thermo-sensitive genic male sterility (TGMS) was induced by employing the CRISPR/Cas9 gene editing technology to modify the gene TMS5. Two TGMS mutants, tms5-1 and tms5-2, both lacking any residual T-DNA, were generated in the indica rice cultivar Zhongjiazao17 (cv. YK17) background. When grown at a sub-optimal temperature (22 °C), both mutants produced viable pollen and successfully produced grain through self-fertilization, but at temperatures 24 and 26 °C, their pollen was sterile and no grain was set. F1 hybrids derived from the crosses between YK17S (tms5-1) and three different restorer lines outperformed both parental lines with respect to grain yield and related traits. CONCLUSION: The YK17S generated by CRISPR/Cas9 system was proved to be a new TGMS line with superior yield potential and can be widely utilized in two-line hybrid breeding of indica rice.


Assuntos
Sistemas CRISPR-Cas , Oryza/genética , Melhoramento Vegetal/métodos , Infertilidade das Plantas/genética , Proteínas de Plantas/genética , Quimera , Mutagênese , Oryza/fisiologia , Plantas Geneticamente Modificadas , Pólen/genética , Pólen/crescimento & desenvolvimento , Temperatura Ambiente
13.
BMC Plant Biol ; 19(Suppl 1): 51, 2019 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-30813888

RESUMO

BACKGROUND: More than 70 cytoplasmic male sterility (CMS) types have been identified in Helianthus, but only for less than half of them, research of mitochondrial organization has been conducted. Moreover, complete mitochondrion sequences have only been published for two CMS sources - PET1 and PET2. It has been demonstrated that other sunflower CMS sources like MAX1, significantly differ from the PET1 and PET2 types. However, possible molecular causes for the CMS induction by MAX1 have not yet been proposed. In the present study, we have investigated structural changes in the mitochondrial genome of HA89 (MAX1) CMS sunflower line in comparison to the fertile mitochondrial genome. RESULTS: Eight significant major reorganization events have been determined in HA89 (MAX1) mtDNA: one 110 kb inverted region, four deletions of 439 bp, 978 bp, 3183 bp and 14,296 bp, respectively, and three insertions of 1999 bp, 5272 bp and 6583 bp. The rearrangements have led to functional changes in the mitochondrial genome of HA89 (MAX1) resulting in the complete elimination of orf777 and the appearance of new ORFs - orf306, orf480, orf645 and orf1287. Aligning the mtDNA of the CMS sources PET1 and PET2 with MAX1 we found some common reorganization features in their mitochondrial genome sequences. CONCLUSION: The new open reading frame orf1287, representing a chimeric atp6 gene, may play a key role in MAX1 CMS phenotype formation in sunflower, while the contribution of other mitochondrial reorganizations seems to appear negligible for the CMS development.


Assuntos
Genoma Mitocondrial/genética , Helianthus/genética , Helianthus/fisiologia , Infertilidade das Plantas/fisiologia , Proteínas de Plantas/metabolismo , Infertilidade das Plantas/genética , Proteínas de Plantas/genética
14.
Gene ; 696: 113-121, 2019 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-30776462

RESUMO

Multiple-allele-inherited male sterility (MAMS) is important in Chinese cabbage (Brassica rapa L. ssp. pekinensis) breeding, but the molecular mechanisms leading to male sterility are poorly understood. In this study, we cloned a novel gene, BrSKS13, that is differentially expressed in fertile and sterile flower buds of Chinese cabbage. BrSKS13 is most similar to Arabidopsis thaliana AT3G13400 (SKS13) and encodes a predicted 61.87 kDa protein with three cupredoxin superfamily conserved domains in the multicopper oxidase family. Semi-quantitative reverse-transcription PCR (sqRT-PCR) showed that expression of BrSKS13 is higher in fertile buds than in sterile buds. Quantitative RT-PCR (qRT-PCR) and in situ hybridization showed that BrSKS13 is highly expressed in fertile anthers, peaking at pollen-maturation stage VI, but is weakly expressed in other tissues and floral organs. Expression patterns of BrSKS13 promoter::GUS reporter fusions in Arabidopsis showed that the BrSKS13 promoter drives expression of the GUS gene only in anthers. The relative expression of Brsks13 in fertile buds was higher than in sterile buds for all other MAMS lines of Chinese cabbage examined. These results suggest that BrSKS13 affects pollen development. In situ hybridization analysis of flower stigmas at different times after pollination showed that BrSKS13 expression was first observed in stigmas and immature seeds at 1 h after pollination, and the signal intensity in seeds increased with increasing maturity. Compared to Col-0, A. thaliana sks13 mutant plants have shorter and fewer siliques, shriveled pollen grains, pollen tube abnormalities, and reduced seed number. The phenotype of sks13 mutant was recovered by over-expressing BrSKS13. Our results suggest that BrSKS13 affects pollen development and the pollination/fertilization process, and will enable further study of the genetic mechanisms underlying MAMS in Chinese cabbage.


Assuntos
Brassica rapa/genética , Regulação da Expressão Gênica no Desenvolvimento/genética , Regulação da Expressão Gênica de Plantas/genética , Infertilidade das Plantas/genética , Proteínas de Plantas/genética , Polinização/genética , Alelos , Arabidopsis/genética , Brassica rapa/crescimento & desenvolvimento , Fertilização/genética , Perfilação da Expressão Gênica , Herança Multifatorial/genética , Plantas Geneticamente Modificadas , Pólen/crescimento & desenvolvimento
15.
Int J Mol Sci ; 20(3)2019 Jan 29.
Artigo em Inglês | MEDLINE | ID: mdl-30699994

RESUMO

Cytoplasmic male sterility (CMS), which is controlled by mitochondrial genes, is an important trait for commercial hybrid seed production. So far, genes controlling this trait are still not clear in pepper. In this study, complete mitochondrial genomes were sequenced and assembled for the CMS line 138A and its maintainer line 138B. The genome size of 138A is 504,210 bp, which is 8618 bp shorter than that of 138B. Meanwhile, more than 214 and 215 open reading frames longer than 100 amino acids (aas) were identified in 138A and 138B, respectively. Mitochondrial genome structure of 138A was quite different from that of 138B, indicating the existence of recombination and rearrangement events. Based on the mitochondrial genome sequence and structure variations, mitochondrion of 138A and FS4401, a Korean origin CMS line, may have inherited from a common female ancestor, but their CMS traits did originate separately. Candidate gene selection was performed according to the published characteristics of the CMS genes, including the presence SNPs and InDels, located in unique regions, their chimeric structure, co-transcription, and transmembrane domain. A total of 35 ORFs were considered as potential candidate genes and 14 of these were selected, with orf300a and 0rf314a as strong candidates. A new marker, orf300a, was developed which did co-segregate with the CMS trait.


Assuntos
Capsicum/genética , Capsicum/fisiologia , Genoma Mitocondrial/genética , Infertilidade das Plantas/genética , Infertilidade das Plantas/fisiologia , Citoplasma/metabolismo , Proteínas de Plantas/genética
16.
Biol Res ; 52(1): 6, 2019 Feb 06.
Artigo em Inglês | MEDLINE | ID: mdl-30728078

RESUMO

BACKGROUND: Pollen development is an energy-consuming process that particularly occurs during meiosis. Low levels of adenosine triphosphate (ATP) may cause cell death, resulting in CMS (cytoplasmic male sterility). DNA sequence differences in ATP synthase genes have been revealed between the N- and S-cytoplasms in the cotton CMS system. However, very few data are available at the RNA level. In this study, we compared five ATP synthase genes in the H276A, H276B and fertile F1 (H276A/H268) lines using RNA editing, RNA blotting and quantitative real time-PCR (qRT-PCR) to explore their contribution to CMS. A molecular marker for identifying male sterile cytoplasm (MSC) was also developed. RESULTS: RNA blotting revealed the absence of any novel orf for the ATP synthase gene sequence in the three lines. Forty-one RNA editing sites were identified in the coding sequences. RNA editing showed that proteins had 32.43% higher hydrophobicity and that 39.02% of RNA editing sites had proline converted to leucine. Two new stop codons were detected in atp6 and atp9 by RNA editing. Real-time qRT-PCR data showed that the atp1, atp6, atp8, and atp9 genes had substantially lower expression levels in H276A compared with those in H276B. By contrast, the expression levels of all five genes were increased in F1 (H276A/H268). Moreover, a molecular marker based on a 6-bp deletion upstream of atp8 in H276A was developed to identify male sterile cytoplasm (MSC) in cotton. CONCLUSIONS: Our data substantially contributes to the understanding of the function of ATP synthase genes in cotton CMS. Therefore, we suggest that ATP synthase genes might be an indirect cause of cotton CMS. Further research is needed to investigate the relationship among ATP synthase genes in cotton CMS.


Assuntos
Adenosina Trifosfatases/genética , Citoplasma/genética , Gossypium/enzimologia , Infertilidade das Plantas/genética , Edição de RNA , Citoplasma/metabolismo , DNA Mitocondrial/genética , Regulação da Expressão Gênica de Plantas/genética , Gossypium/genética , Reação em Cadeia da Polimerase , RNA Mitocondrial/genética
17.
BMC Plant Biol ; 19(1): 65, 2019 Feb 11.
Artigo em Inglês | MEDLINE | ID: mdl-30744566

RESUMO

BACKGROUND: Thermo-sensitive genetic male sterile (TGMS) lines have been widely used in two-line hybrid rice breeding. The two-line hybrids have increased rice yields substantially. However, the effect of environmental temperatures on the fertility conversion is still not fully clear. In this study, we performed a tandem mass tag (TMT)-based proteomic analysis on the anthers of the TGMS line AnnongS-1 grown under permissive (low) temperature (21 °C) and restrictive (high) temperature (> 26 °C) conditions in an attempt to explore the effect of temperature on the fertility of the male sterile line. RESULTS: After the AnnongS-1 plants were induced under either permissive or restrictive conditions, morphological observations and I2-KI staining confirmed that the pollen grains formed under high temperature conditions were abortive while those formed under low temperature developed normally. In comparison to the plants grown under permissive conditions, the restrictive high-temperature conditions led to the differential accumulation of 89 proteins in the anthers, of which 46 were increased in abundance and 43 were decreased in abundance. Most of the subcellular compartments of the anther cells had one or more proteins that had been differentially accumulated, with the cytoplasm and chloroplast having the greatest accumulations. More than 40% of the differentially abundant proteins (DAPs) were enzymes involved in photosynthesis, energy metabolism, biosynthesis and catabolism of cellular components, metabolic regulation, defense and stress, etc. The DAPs related to protein metabolism accounted for the largest proportion (21.35%), followed by those related to defense and stress (12.36%), metabolic regulation (10.11%) and carbohydrate metabolism (8.99%), indicating that such biological processes in anther cells were more susceptible to high temperature stress. CONCLUSIONS: The restrictive temperature induction caused fertility-sterility conversion in the TGMS line AnnongS-1 mainly by adversely affecting the metabolism of protein, carbohydrate and energy, and decreasing the abundances of important proteins closely related to defense and stress, thereby impeding the growth and development of the pollen and weakening the overall defense and ability to endure stress of AnnongS-1. These data are helpful for deepening our understanding of the molecular mechanism underlying fertility conversion in TGMS lines.


Assuntos
Infertilidade das Plantas/fisiologia , Proteômica/métodos , Oryza/genética , Oryza/metabolismo , Melhoramento Vegetal , Infertilidade das Plantas/genética , Temperatura Ambiente
18.
Methods Mol Biol ; 1917: 97-107, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30610631

RESUMO

Obtaining photoperiod-sensitive genic male sterility (PGMS) lines is one of the most important steps in two-line hybrid rice breeding. Traditionally, such lines were screened and developed with a classic rice breeding system under both long-day and short-day conditions. The isolation and backcross process used for this could easily last for more than 3 years with a very low success rate. Here, we describe a straightforward method for generating csa-based PGMS lines by using the CRISPR-Cas9 technology in rice.


Assuntos
Sistemas CRISPR-Cas/genética , Oryza/genética , Oryza/fisiologia , Fotoperíodo , Infertilidade das Plantas/fisiologia , Melhoramento Vegetal , Infertilidade das Plantas/genética
19.
Int J Mol Sci ; 19(12)2018 Dec 12.
Artigo em Inglês | MEDLINE | ID: mdl-30545137

RESUMO

In flowering plants, ideal male reproductive development requires the systematic coordination of various processes, in which timely differentiation and degradation of the anther wall, especially the tapetum, is essential for both pollen formation and anther dehiscence. Here, we show that OsGPAT3, a conserved glycerol-3-phosphate acyltransferase gene, plays a critical role in regulating anther wall degradation and pollen exine formation. The gpat3-2 mutant had defective synthesis of Ubisch bodies, delayed programmed cell death (PCD) of the inner three anther layers, and abnormal degradation of micropores/pollen grains, resulting in failure of pollen maturation and complete male sterility. Complementation and clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) experiments demonstrated that OsGPAT3 is responsible for the male sterility phenotype. Furthermore, the expression level of tapetal PCD-related and nutrient metabolism-related genes changed significantly in the gpat3-2 anthers. Based on these genetic and cytological analyses, OsGPAT3 is proposed to coordinate the differentiation and degradation of the anther wall and pollen grains in addition to regulating lipid biosynthesis. This study provides insights for understanding the function of GPATs in regulating rice male reproductive development, and also lays a theoretical basis for hybrid rice breeding.


Assuntos
Apoptose , Oryza/citologia , Oryza/metabolismo , Proteínas de Plantas/metabolismo , Pólen/citologia , Pólen/crescimento & desenvolvimento , Sequência de Bases , Mapeamento Cromossômico , Fragmentação do DNA , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Estudos de Associação Genética , Teste de Complementação Genética , Mutação/genética , Oryza/genética , Fenótipo , Infertilidade das Plantas/genética , Proteínas de Plantas/genética , Pólen/metabolismo , Pólen/ultraestrutura , Reprodutibilidade dos Testes
20.
Int J Mol Sci ; 19(12)2018 Dec 12.
Artigo em Inglês | MEDLINE | ID: mdl-30545163

RESUMO

The interaction between plant mitochondria and the nucleus markedly influences stress responses and morphological features, including growth and development. An important example of this interaction is cytoplasmic male sterility (CMS), which results in plants producing non-functional pollen. In current research work, we compared the phenotypic differences in floral buds of different Brassica napus CMS (Polima, Ogura, Nsa) lines with their corresponding maintainer lines. By comparing anther developmental stages between CMS and maintainer lines, we identified that in the Nsa CMS line abnormality occurred at the tetrad stage of pollen development. Phytohormone assays demonstrated that IAA content decreased in sterile lines as compared to maintainer lines, while the total hormone content was increased two-fold in the S2 stage compared with the S1 stage. ABA content was higher in the S1 stage and exhibited a two-fold decreasing trend in S2 stage. Sterile lines however, had increased ABA content at both stages compared with the corresponding maintainer lines. Through transcriptome sequencing, we compared differentially expressed unigenes in sterile and maintainer lines at both (S1 and S2) developmental stages. We also explored the co-expressed genes of the three sterile lines in the two stages and classified these genes by gene function. By analyzing transcriptome data and validating by RT-PCR, it was shown that some transcription factors (TFs) and hormone-related genes were weakly or not expressed in the sterile lines. This research work provides preliminary identification of the pollen abortion stage in Nsa CMS line. Our focus on genes specifically expressed in sterile lines may be useful to understand the regulation of CMS.


Assuntos
Brassica napus/genética , Citoplasma/metabolismo , Reguladores de Crescimento de Planta/metabolismo , Infertilidade das Plantas/genética , Transcriptoma/genética , Flores/anatomia & histologia , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Ontologia Genética , Genes de Plantas , Fenótipo , Reprodutibilidade dos Testes
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