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1.
Biochem Biophys Res Commun ; 733: 150615, 2024 Aug 28.
Artigo em Inglês | MEDLINE | ID: mdl-39213704

RESUMO

Fruit ripening is a highly intricate process, where the dynamic interplay of soluble sugar and organic acid metabolism is crucial for developing the characteristic flavor qualities. Pyruvate orthophosphate dikinase (PPDK) plays a pivotal role in modulating the process of gluconeogenesis during plant development. However, the specific physiological role of PPDK in fruit development has yet to be elucidated. In this study, we investigated the expression pattern, subcellular localization and functional significance of SlPPDK in tomato fruits. Our results reveal that SlPPDK is highly expressed in fruits and flowers, with its expression progressively increasing as the fruit ripens. Subcellular localization analyses demonstrate that SlPPDK is distributed in the cell membrane, cytoplasm, and nucleus. Using CRISPR/Cas9 technology, we generated SlPPDK knockout mutants, which exhibited a marked reduction in enzyme activity, leading to significant alterations in sugar and organic acid metabolism. These findings highlight the critical role of SlPPDK in maintaining the sugar-acid balance essential for tomato flavor quality and provide a foundation for future breeding strategies aimed at enhancing tomato fruit flavor.

2.
Int J Mol Sci ; 24(23)2023 Nov 28.
Artigo em Inglês | MEDLINE | ID: mdl-38069184

RESUMO

The membrane-less organelles in cytoplasm that are presented as cytoplasmic foci were successively identified. Although multiple CCCH zinc-finger proteins have been found to be localized in cytoplasmic foci, the relationship between their specific localization and functions still needs further clarification. Here, we report that the heterologous expression of two Brassica campestris CCCH zinc-finger protein genes (BcMF30a and BcMF30c) in Arabidopsis thaliana can affect microgametogenesis by involving the formation of cytoplasmic foci. By monitoring the distribution of proteins and observing pollen phenotypes, we found that, when these two proteins were moderately expressed in pollen, they were mainly dispersed in the cytoplasm, and the pollen developed normally. However, high expression induced the assembly of cytoplasmic foci, leading to pollen abortion. These findings suggested that the continuous formation of BcMF30a/BcMF30c-associated cytoplasmic foci due to high expression was the inducement of male sterility. A co-localization analysis further showed that these two proteins can be recruited into two well-studied cytoplasmic foci, processing bodies (PBs), and stress granules (SGs), which were confirmed to function in mRNA metabolism. Together, our data suggested that BcMF30a and BcMF30c play component roles in the assembly of pollen cytoplasmic foci. Combined with our previous study on the homologous gene of BcMF30a/c in Arabidopsis, we concluded that the function of these homologous genes is conserved and that cytoplasmic foci containing BcMF30a/c may participate in the regulation of gene expression in pollen by regulating mRNA metabolism.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Brassica , Arabidopsis/genética , Arabidopsis/metabolismo , Brassica/genética , Brassica/metabolismo , Proteínas de Arabidopsis/genética , Pólen/genética , Pólen/metabolismo , RNA Mensageiro/metabolismo , Zinco/metabolismo , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/metabolismo , Dedos de Zinco/genética
3.
Int J Mol Sci ; 24(23)2023 Nov 24.
Artigo em Inglês | MEDLINE | ID: mdl-38069011

RESUMO

Cruciferous plants manufacture glucosinolates (GSLs) as special and important defense compounds against insects. However, how insect feeding induces glucosinolates in Brassica to mediate insect resistance, and how plants regulate the strength of anti-insect defense response during insect feeding, remains unclear. Here, mustard (Brassica juncea), a widely cultivated Brassica plant, and beet armyworm (Spodoptera exigua), an economically important polyphagous pest of many crops, were used to analyze the changes in GSLs and transcriptome of Brassica during insect feeding, thereby revealing the plant-insect interaction in Brassica plants. The results showed that the content of GSLs began to significantly increase after 48 h of herbivory by S. exigua, with sinigrin as the main component. Transcriptome analysis showed that a total of 8940 DEGs were identified in mustard challenged with beet armyworm larvae. The functional enrichment results revealed that the pathways related to the biosynthesis of glucosinolate and jasmonic acid were significantly enriched by upregulated DEGs, suggesting that mustard might provide a defense against herbivory by inducing JA biosynthesis and then promoting GSL accumulation. Surprisingly, genes regulating JA catabolism and inactivation were also activated, and both JA signaling repressors (JAZs and JAMs) and activators (MYCs and NACs) were upregulated during herbivory. Taken together, our results indicate that the accumulation of GSLs regulated by JA signaling, and the regulation of active and inactive JA compound conversion, as well as the activation of JA signaling repressors and activators, collectively control the anti-insect defense response and avoid over-stunted growth in mustard during insect feeding.


Assuntos
Beta vulgaris , Mostardeira , Animais , Mostardeira/genética , Mostardeira/metabolismo , Transcriptoma , Spodoptera/fisiologia , Glucosinolatos/metabolismo , Beta vulgaris/genética , Beta vulgaris/metabolismo , Herbivoria/genética , Insetos/metabolismo
4.
Int J Mol Sci ; 23(11)2022 Jun 02.
Artigo em Inglês | MEDLINE | ID: mdl-35682925

RESUMO

The development of flower and pollen is a complex biological process that involves multiple metabolic pathways in plants. In revealing novel insights into flower and pollen development underlying male sterility (MS), we conducted an integrated profiling of gene and protein activities in developing buds in cytoplasmic male sterile (CMS) mutants of mustard (Brassica juncea). Using RNA-Seq and label-free quantitative proteomics, 11,832 transcripts and 1780 protein species were identified with significant differential abundance between the male sterile line 09-05A and its maintainer line 09-05B at the tetrad stage and bi-nucleate stage of B. juncea. A large number of differentially expressed genes (DEGs) and differentially abundant proteins (DAPs) involved in carbohydrate and energy metabolism, including starch and sucrose metabolism, tricarboxylic acid (TCA) cycle, glycolysis, and oxidoreductase activity pathways, were significantly downregulated in 09-05A buds. The low expression of these DEGs or functional loss of DAPs, which can lead to an insufficient supply of critical substrates and ATP, could be associated with flower development, pollen development, and changes in fertility in B. juncea. Therefore, this study provided transcriptomic and proteomic information of pollen abortion for B. juncea and a basis for further research on the molecular regulatory mechanism of MS in plants.


Assuntos
Infertilidade Masculina , Mostardeira , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Humanos , Masculino , Mostardeira/genética , Infertilidade das Plantas/genética , Proteínas de Plantas/genética , Proteômica , Transcriptoma
5.
BMC Plant Biol ; 21(1): 254, 2021 Jun 03.
Artigo em Inglês | MEDLINE | ID: mdl-34082704

RESUMO

BACKGROUND: As an important subfamily of arabinogalactan proteins (AGPs), fasciclin-like AGPs (FLAs) contribute to various aspects of growth, development and adaptation, yet their function remains largely elusive. Despite the diversity of FLAs, only two members, Arabidopsis FLA3 and rice MTR1, are reported to be involved in sexual reproduction. In this study, another Arabidopsis FLA-encoding gene, FLA14, was identified, and its role was investigated. RESULTS: Arabidopsis FLA14 was found to be a pollen grain-specific gene. Expression results from fusion with green fluorescent protein showed that FLA14 was localized along the cell membrane and in Hechtian strands. A loss-of-function mutant of FLA14 showed no discernible defects during male gametogenesis, but precocious pollen germination occurred inside the mature anthers under high moisture conditions. Overexpression of FLA14 caused 39.2% abnormal pollen grains with a shrunken and withered appearance, leading to largely reduced fertility with short mature siliques and lower seed set. Cytological and ultramicroscopic observation showed that ectopic expression of FLA14 caused disruption at the uninucleate stage, resulting in either collapsed pollen with absent intine or pollen of normal appearance but with a thickened intine. CONCLUSIONS: Taken together, our data suggest a role for FLA14 in pollen development and preventing premature pollen germination inside the anthers under high relative humidity in Arabidopsis.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Proteínas Ligadas por GPI/metabolismo , Regulação da Expressão Gênica de Plantas/fisiologia , Pólen/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Membrana Celular , Proteínas Ligadas por GPI/genética , Plantas Geneticamente Modificadas , Pólen/genética , Transporte Proteico , Água
6.
Biochem Biophys Res Commun ; 528(1): 140-145, 2020 07 12.
Artigo em Inglês | MEDLINE | ID: mdl-32451083

RESUMO

The membraneless messenger ribonucleoprotein (mRNP) granules, including processing bodies (PBs) and stress granules (SGs), are important cytoplasmic structures in eukaryotes that can participate in gene expression through mRNA regulation. It has been verified that mRNP granules are mainly composed of proteins and translation-repressed mRNAs. Here, we reported a stop-codon read-through gene, At3g52980, in plants for the first time. At3g52980 encodes a novel non-tandem CCCH zinc-finger (non-TZF) protein named AtC3H18-Like (AtC3H18L), which contains two putative RNA-binding domains. By using transient expression system, we showed that heat treatment can induce the aggregation of diffuse distributed AtC3H18L to form cytoplasmic foci, which were similar to PBs and SGs in morphology. Further analysis did find that AtC3H18L can co-localize with markers of PB and SG. The aggregation of AtC3H18L was closely related to the cytoskeleton, and AtC3H18L-foci were highly dynamic and can move frequently along cytoskeleton. Moreover, analysis in transgenic plants showed that AtC3H18L was specifically expressed in pollen and can form cytoplasmic foci without heat treatment. It will be fascinating in future studies to discover whether and how AtC3H18L affects pollen development by participating in the assembly of mRNP granules as a protein component, especially under heat stress.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Códon de Terminação/genética , Grânulos Citoplasmáticos/metabolismo , Proteínas de Ligação a RNA/metabolismo , Ribonucleoproteínas/metabolismo , Dedos de Zinco , Proteínas de Arabidopsis/química , Proteínas de Arabidopsis/genética , Regulação da Expressão Gênica de Plantas , Resposta ao Choque Térmico , Inflorescência/metabolismo , Epiderme Vegetal/citologia , Plantas Geneticamente Modificadas , Pólen/metabolismo , Domínios Proteicos , Proteínas de Ligação a RNA/química , Proteínas de Ligação a RNA/genética , Frações Subcelulares/metabolismo , Nicotiana/genética
7.
Int J Mol Sci ; 21(17)2020 Sep 03.
Artigo em Inglês | MEDLINE | ID: mdl-32899329

RESUMO

Chinese cabbage (Brassica campestris) is an economically important leaf vegetable crop worldwide. Mounting studies have shown that cysteine-cysteine-cysteine-histidine (CCCH) zinc-finger protein genes are involved in various plant growth and development processes. However, research on the involvement of these genes in male reproductive development is still in its infancy. Here, we identified 11 male fertility-related CCCH genes in Chinese cabbage. Among them, a pair of paralogs encoding novel non-tandem CCCH zinc-finger proteins, Brassica campestris Male Fertility 30a (BcMF30a) and BcMF30c, were further characterized. They were highly expressed in pollen during microgametogenesis and continued to express in germinated pollen. Further analyses demonstrated that both BcMF30a and BcMF30c may play a dual role as transcription factors and RNA-binding proteins in plant cells. Functional analysis showed that partial bcmf30a bcmf30c pollen grains were aborted due to the degradation of pollen inclusion at the microgametogenesis phase, and the germination rate of viable pollen was also greatly reduced, indicating that BcMF30a and BcMF30c are required for both pollen development and pollen germination. This research provided insights into the function of CCCH proteins in regulating male reproductive development and laid a theoretical basis for hybrid breeding of Chinese cabbage.


Assuntos
Brassica/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas , Germinação , Proteínas de Plantas/metabolismo , Pólen/crescimento & desenvolvimento , Dedos de Zinco , Brassica/metabolismo , Proteínas de Plantas/genética , Pólen/metabolismo
8.
Plant J ; 94(1): 60-76, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29385650

RESUMO

Arabinogalactan proteins (AGPs) are extensively glycosylated hydroxyproline-rich glycoproteins ubiquitous in all plant tissues and cells. AtAGP6 and AtAGP11, the only two functionally known pollen-specific classical AGP encoding genes in Arabidopsis, are reported to have redundant functions in microspore development. BcMF18 and BcMF8 isolated from Brassica campestris are the orthologues of AtAGP6 and AtAGP11, respectively. In contrast to the functional redundancy of AtAGP6 and AtAGP11, single-gene disruption of BcMF8 led to deformed pollen grains with abnormal intine development and ectopic aperture formation in B. campestris. Here, we further explored the action of BcMF18 and its relationship with BcMF8. BcMF18 was specifically expressed in pollen during the late stages of microspore development. Antisense RNA transgenic lines with BcMF18 reduction resulted in aberrant pollen grains with abnormal cellulose distribution, lacking intine, cytoplasm and nuclei. Transgenic plants with repressive expression of both BcMF8 and BcMF18 showed a hybrid phenotype, expressing a mixture of the phenotypes of the single gene knockdown plant lines. In addition, we identified functional diversity between BcMF18/BcMF8 and AtAGP6/AtAGP11, mainly reflected by the specific contribution of BcMF18 and BcMF8 to pollen wall formation. These results suggest that, unlike the orthologous genes AtAGP6 and AtAGP11 in Arabidopsis, BcMF18 and BcMF8 are both integral to pollen biogenesis in B. campestris, acting through independent pathways during microspore development.


Assuntos
Brassica/crescimento & desenvolvimento , Galactanos/metabolismo , Glicoproteínas/fisiologia , Proteínas de Plantas/fisiologia , Pólen/crescimento & desenvolvimento , Brassica/metabolismo , Técnicas de Silenciamento de Genes , Glicoproteínas/metabolismo , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Pólen/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Alinhamento de Sequência
9.
Plant Cell Rep ; 37(7): 1003-1009, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29644403

RESUMO

KEY MESSAGE: BcMF23a contributes to pollen wall development via influencing intine construction, which, in turn, influences pollen tube growth. Pollen wall, the morphological out face of pollen, surrounds male gametophyte and plays an important role in plant reproduction. Pectin methylesterases (PMEs) are involved in pollen wall construction by de-esterifying pectin of the intine. In this study, the function of a putative pectin methylesterase gene, Brassica campestris Male Fertility 23a (BcMF23a), was investigated. Knockdown of BcMF23a by artificial microRNA (amiRNA) technology resulted in abnormal pollen intine formation outside of the germinal furrows at the binucleate stage. At the trinucleate stage, 20.69% of pollen possessed the degradation of nuclei, cytoplasm and the intine, resulting in shrunken pollen, whereas the remaining 75.86% were wall-disrupted with degrading cytoplasm and broken exine inside the germinal furrows. In addition, pollen abortion in transgenic plants caused germination percentage reduction by 19% in vitro and pollen tube growth disruption in natural stigma in vivo. Taken together, BcMF23a is involved in pollen development and pollen tube growth, possibly via participating in intine construction. This study may contribute towards understanding the function of pollen-specific PMEs and the molecular regulatory network of pollen wall development.


Assuntos
Brassica/genética , Hidrolases de Éster Carboxílico/genética , Proteínas de Plantas/genética , Tubo Polínico/crescimento & desenvolvimento , Brassica/crescimento & desenvolvimento , Hidrolases de Éster Carboxílico/metabolismo , Regulação da Expressão Gênica de Plantas , Técnicas de Silenciamento de Genes , Germinação , MicroRNAs , Microscopia Eletrônica de Varredura , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Tubo Polínico/genética , Tubo Polínico/ultraestrutura
10.
Int J Mol Sci ; 19(5)2018 May 02.
Artigo em Inglês | MEDLINE | ID: mdl-29724020

RESUMO

Pectin methylesterase inhibitor genes (PMEIs) are a large multigene family and play crucial roles in cell wall modifications in plant growth and development. Here, a comprehensive analysis of the PMEI gene family in Brassicacampestris, an important leaf vegetable, was performed. We identified 100 BrassicacampestrisPMEI genes (BcPMEIs), among which 96 BcPMEIs were unevenly distributed on 10 chromosomes and nine tandem arrays containing 20 BcPMEIs were found. We also detected 80 pairs of syntenic PMEI orthologs. These findings indicated that whole-genome triplication (WGT) and tandem duplication (TD) were the main mechanisms accounting for the current number of BcPMEIs. In evolution, BcPMEIs were retained preferentially and biasedly, consistent with the gene balance hypothesis and two-step theory, respectively. The molecular evolution analysis of BcPMEIs manifested that they evolved through purifying selection and the divergence time is in accordance with the WGT data of B. campestris. To obtain the functional information of BcPMEIs, the expression patterns in five tissues and the cis-elements distributed in promoter regions were investigated. This work can provide a better understanding of the molecular evolution and biological function of PMEIs in B. campestris.


Assuntos
Brassica/enzimologia , Hidrolases de Éster Carboxílico/metabolismo , Evolução Molecular , Perfilação da Expressão Gênica , Estudo de Associação Genômica Ampla , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Brassica/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas , Genoma de Planta/genética , Filogenia , Proteínas de Plantas/classificação , Análise de Sequência de DNA , Análise de Sequência de Proteína , Sintenia , Sequências de Repetição em Tandem/genética
11.
Int J Mol Sci ; 19(11)2018 Oct 26.
Artigo em Inglês | MEDLINE | ID: mdl-30373125

RESUMO

The activities of pectin methylesterases (PMEs) are regulated by pectin methylesterase inhibitors (PMEIs), which consequently control the pectin methylesterification status. However, the role of PMEI genes in Brassica oleracea, an economically important vegetable crop, is poorly understood. In this study, 95 B. oleracea PMEI (BoPMEI) genes were identified. A total of 77 syntenic ortholog pairs and 10 tandemly duplicated clusters were detected, suggesting that the expansion of BoPMEI genes was mainly attributed to whole-genome triplication (WGT) and tandem duplication (TD). During diploidization after WGT, BoPMEI genes were preferentially retained in accordance with the gene balance hypothesis. Most homologous gene pairs experienced purifying selection with ω (Ka/Ks) ratios lower than 1 in evolution. Five stamen-specific BoPMEI genes were identified by expression pattern analysis. By combining the analyses of expression and evolution, we speculated that nonfunctionalization, subfunctionalization, neofunctionalization, and functional conservation can occur in the long evolutionary process. This work provides insights into the characterization of PMEI genes in B. oleracea and contributes to the further functional studies of BoPMEI genes.


Assuntos
Brassica/genética , Hidrolases de Éster Carboxílico/genética , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Proteínas de Plantas/genética , Brassica/enzimologia , Diploide , Evolução Molecular , Duplicação Gênica , Família Multigênica , Transcriptoma
12.
Int J Mol Sci ; 19(8)2018 Aug 04.
Artigo em Inglês | MEDLINE | ID: mdl-30081560

RESUMO

Polygalacturonase (PG), a large hydrolase family in plants, is involved in pectin disassembly of the cell wall in plants. The present study aims to characterize PG genes and investigate their expression patterns in Solanum lycopersicum. We identified 54 PG genes in the tomato genome and compared their amino acid sequences with their Arabidopsis counterpart. Subsequently, we renamed these PG genes according to their Arabidopsis homologs. Phylogenetic and evolutionary analysis revealed that these tomato PG genes could be classified into seven clades, and within each clade the exon/intron structures were conserved. Expression profiles analysis through quantitive real-time polymerase chain reaction (qRT-PCR) revealed that most SlPGs had specific or high expression patterns in at least one organ, and particularly five PG genes (SlPG14, SlPG15, SlPG49, SlPG70, and SlPG71) associated with fruit development. Promoter analysis showed that more than three cis-elements associated with plant hormone response, environmental stress response or specific organ/tissue development exhibited in each SlPG promoter regions. In conclusion, our results may provide new insights for the further study of PG gene function during plant development.


Assuntos
Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Poligalacturonase/metabolismo , Solanum lycopersicum/enzimologia , Solanum lycopersicum/genética , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Genoma de Planta/genética , Filogenia , Proteínas de Plantas/classificação , Poligalacturonase/genética
13.
Funct Integr Genomics ; 16(6): 641-656, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27632199

RESUMO

Cell walls are a distinguishing characteristic of plants essential to their survival. The pectin content of primary cell walls in grasses and dicots is distinctly different. Polygalacturonases (PGs) can degrade pectins and participate in multiple developmental processes of plants. This study comprehensively compared the evolution, expression, and cis-regulatory element of PGs in grasses and dicots. A total of 577 PGs identified from five grasses and five dicots fell into seven clades. Evolutionary analysis demonstrated the distinct differences between grasses and dicots in patterns of gene duplication and loss, and evolutionary rates. Grasses generally contained much fewer clade C and F members than dicots. We found that this disparity was the result of less duplication and more gene losses in grasses. More duplications occurred in clades D and E, and expression analysis showed that most of clade E members were expressed ubiquitously at a high overall level and clade D members were closely related to male reproduction in both grasses and dicots, suggesting their biological functions were highly conserved across species. In addition to the general role in reproductive development, PGs of clades C and F specifically played roles in root development in dicots, shedding light on organ differentiation between the two groups of plants. A regulatory element analysis of clade C and F members implied that possible functions of PGs in specific biological responses contributed to their expansion and preservation. This work can improve the knowledge of PGs in plants generally and in grasses specifically and is beneficial to functional studies.


Assuntos
Evolução Molecular , Pectinas/metabolismo , Poaceae/genética , Poligalacturonase/genética , Parede Celular/genética , Parede Celular/metabolismo , Regulação da Expressão Gênica de Plantas , Magnoliopsida/genética , Pectinas/genética , Filogenia , Raízes de Plantas/genética , Raízes de Plantas/crescimento & desenvolvimento , Poaceae/classificação , Poligalacturonase/biossíntese , Sequências Reguladoras de Ácido Nucleico/genética
14.
Plant Mol Biol ; 89(6): 629-46, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26506823

RESUMO

Polygalacturonases (PGs) participate in pectin disassembly of cell wall and belong to one of the largest hydrolase families in plants. In this study, we identified 99 PG genes in Brassica rapa. Comprehensive analysis of phylogeny, gene structures, physico-chemical properties and coding sequence evolution demonstrated that plant PGs should be classified into seven divergent clades and each clade's members had specific sequence and structure characteristics, and/or were under specific selection pressures. Genomic distribution and retention rate analysis implied duplication events and biased retention contributed to PG family's expansion. Promoter divergence analysis using "shared motif method" revealed a significant correlation between regulatory and coding sequence evolution of PGs, and proved Clades A and E were of ancient origin. Quantitative real-time PCR analysis showed that expression patterns of PGs displayed group specificities in B. rapa. Particularly, nearly half of PG family members, especially those of Clades C, D and F, closely relates to reproductive development. Most duplicates showed similar expression profiles, suggesting dosage constraints accounted for preservation after duplication. Promoter-GUS assay further indicated PGs' extensive roles and possible redundancy during reproductive development. This work can provide a scientific classification of plant PGs, dissect the internal relationships between their evolution and expressions, and promote functional researches.


Assuntos
Brassica rapa/enzimologia , Brassica rapa/genética , Genes de Plantas , Proteínas de Plantas/genética , Poligalacturonase/genética , Brassica rapa/classificação , Mapeamento Cromossômico , Evolução Molecular , Duplicação Gênica , Regulação Enzimológica da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Modelos Genéticos , Família Multigênica , Filogenia , Plantas Geneticamente Modificadas , Regiões Promotoras Genéticas , Fatores de Tempo
15.
Mol Biol Rep ; 41(1): 537-44, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-24323195

RESUMO

Brassica campestris Male Fertility 21 (BcMF21) was previously isolated from the flower buds of Chinese cabbage (Brassica campestris L. ssp. chinensis Makino, syn. B. rapa ssp. chinensis) and expressed specifically in tapetum and microspores during the meiosis stage and the uninucleate stage of microspore development. Here, we used antisense RNA technology to knock down the expression level of BcMF21 in B. campestris and analyzed the phenotype of the transgenic plants. Alexander staining and scanning electron microscope revealed sterility and exine deformities in the mature pollen grains of BcMF21 antisense RNA transgenic plants. The germination furrow of the BcMF21 antisense RNA transgenic pollen was covered by lipid like materials. The pollen tubes burst and could not grow normally in vitro. Therefore, we presented here BcMF21 might be an important gene for pollen development and germination.


Assuntos
Brassica rapa/crescimento & desenvolvimento , Germinação , Proteínas de Plantas/fisiologia , Pólen/crescimento & desenvolvimento , Brassica rapa/metabolismo , Técnicas de Silenciamento de Genes , Infertilidade das Plantas/genética , Pólen/metabolismo , RNA Antissenso/genética
16.
J Integr Plant Biol ; 56(11): 1095-105, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-24773757

RESUMO

PECTATE LYASE-LIKE10 (PLL10) was previously identified as one of the differentially expressed genes both in microspores during the late pollen developmental stages and in pistils during the fertilization process in Chinese cabbage (Brassica campestris ssp. chinensis). Here, antisense-RNA was used to study the functions of BcPLL10 in Chinese cabbage. Abnormal pollen was identified in the transgenic lines (bcpll10-4, -5, and -6). In fertilization experiments, fewer seeds were harvested when the antisense-RNA lines were used as pollen donor. In vivo and in vitro pollen germination assays less germinated pollen tubes were observed in bcpll10 lines. Scanning electron microscopy observation verified that the tryphine materials were over accumulated around the pollen surface and sticked them together in bcpll10. Moreover, transmission electron microscopy observation revealed that the internal endintine was overdeveloped and predominantly occupied the intine, and disturbed the normal proportional distribution of the two layers in the non-germinal furrow region; and no obvious demarcation existed between them in the germinal furrow region in the bcpll10 pollen. Collectively, this study presented a novel PLL gene that played an important role during the pollen wall development in B. campestris, which may also possess potential importance for male sterility usage in agriculture.


Assuntos
Brassica/enzimologia , Brassica/crescimento & desenvolvimento , Proteínas de Plantas/metabolismo , Pólen/crescimento & desenvolvimento , Polissacarídeo-Liases/metabolismo , Brassica/genética , Brassica/ultraestrutura , Regulação da Expressão Gênica de Plantas , Germinação , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Pólen/ultraestrutura , Polissacarídeo-Liases/genética , RNA Antissenso/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sementes/genética , Sementes/crescimento & desenvolvimento
17.
Mitochondrial DNA B Resour ; 8(10): 1049-1053, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37810612

RESUMO

Plants of the genus Plectranthus are used for the treatment of digestive problems, skin diseases, and allergies, with a wide variety of uses. Here, the complete chloroplast genome sequence of Plectranthus hadiensis (Benth. ex E.Mey) Codd. 1788 was assembled and characterized for the first time. The full length of the chloroplast genome is 152,484 bp, consisting of a small single-copy region of 17,686 bp, a large single-copy region of 83,380 bp, and a pair of inverted repeats of 51,418 bp. The overall GC content is 37.73%. The chloroplast genome contains 131 unique genes, including 87 protein-coding genes, 36 transfer RNA genes, and eight ribosomal RNA genes. Phylogenetic tree construction based on the complete chloroplast genome sequences of 25 species (23 Nepetoideae, two Ajugoideae) of the Lamiaceae family showed that P. hadiensis exhibited the closest relationship with Isodon.

18.
Front Plant Sci ; 13: 932793, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35909782

RESUMO

Plant CCCH zinc-finger proteins form a large family of regulatory proteins function in many aspects of plant growth, development and environmental responses. Despite increasing reports indicate that many CCCH zinc-finger proteins exhibit similar subcellular localization of being localized in cytoplasmic foci, the underlying molecular mechanism and the connection between this specific localization pattern and protein functions remain largely elusive. Here, we identified another cytoplasmic foci-localized CCCH zinc-finger protein, AtC3H18, in Arabidopsis thaliana. AtC3H18 is predominantly expressed in developing pollen during microgametogenesis. Although atc3h18 mutants did not show any abnormal phenotype, possibly due to redundant gene(s), aberrant AtC3H18 expression levels caused by overexpression resulted in the assembly of AtC3H18-positive granules in a dose-dependent manner, which in turn led to male sterility phenotype, highlighting the importance of fine-tuned AtC3H18 expression. Further analyzes demonstrated that AtC3H18-positive granules are messenger ribonucleoprotein (mRNP) granules, since they can exhibit liquid-like physical properties, and are associated with another two mRNP granules known as processing bodies (PBs) and stress granules (SGs), reservoirs of translationally inhibited mRNAs. Moreover, the assembly of AtC3H18-positive granules depends on mRNA availability. Combined with our previous findings on the AtC3H18 homologous genes in Brassica campestris, we concluded that appropriate expression level of AtC3H18 during microgametogenesis is essential for normal pollen development, and we also speculated that AtC3H18 may act as a key component of mRNP granules to modulate pollen mRNAs by regulating the assembly/disassembly of mRNP granules, thereby affecting pollen development.

19.
Mitochondrial DNA B Resour ; 6(9): 2693-2694, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34435122

RESUMO

Glebionis coronaria (Asteraceae) is widely distributed in China, and it regulates the stomach, strengthens the spleen, reduces blood pressure, and reinforces the brain. In this study, the complete chloroplast genome sequence of G. coronaria was reported. The total chloroplast genome cycle was 149,750 bp, and it formed a large single-copy (LSC, 82,290 bp), a small single-copy (SSC, 18,414 bp), and two inverted repeats (IR, 24,523 bp) regions. The GC content of this genome was 36.35%. The whole-genome contained 128 genes, including 84 protein-coding genes, 36 tRNA genes, and eight rRNA genes. Phylogenetic analysis indicated that G. coronaria appeared within a clade comprised of Chrysanthemum species.

20.
Front Plant Sci ; 12: 707054, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34539701

RESUMO

MYC2/3/4, known as a basic helix-loop-helix (bHLH) transcription factor, directly activate the genes involved in diverse plant development and secondary metabolites biosynthesis. In this study, we identified and cloned five MYC paralogs (BrMYC2/3-1/3-2/4-1/4-2) from Chinese cabbage (Brassica rapa ssp. pekinensis). In-silico analyses for the physicochemical properties suggested that BrMYC2/3-1/3-2/4-2/4-3 are unstable hydrophobic and acidic proteins, while BrMYC4-1 is an unstable hydrophobic and basic protein. BrMYC2/3/4 belong to the bHLH superfamily and are closely related to AthMYC2/3/4 orthologs that mediate the regulation of various secondary metabolites. It was demonstrated that BrMYC2/3/4-GFP fusion protein localized in the nucleus and expression levels of five BrMYC2/3/4 homologous genes all elevated relative to control (Ctrl). When expressed in Arabidopsis under the control of 35S promoter, each of the BrMYC2/3-1/3-2/4-1/4-2 transgenes differentially influenced root and shoot elongation, vegetative phase change, flowering time, plant height and tiller number after flowering, and seed production. Despite the variation of phenotypes between the transgenic lines, all the lines except for BrMYC4-2 exhibited shorter seed length, less seed weight, higher accumulation of glucosinolates (GSs), and resistance to Sclerotinia sclerotiorum than Ctrl. Notably, BrMYC2 overexpression (OE) line significantly reduced the lengths of root and hypocotyl, seed length, and weight, along with faster bolting time and strikingly higher accumulation of total GSs. Accumulation of GSs at the highest levels in the BrMYC2 OE line conferred the highest resistance to S. sclerotiorum. Unlike BrMYC3 OE and BrMYC4 OE , BrMYC2 OE stimulated the growth of plant height after fluorescence. The results of this study point to the BrMYC2 overexpression that may provide a beneficial effect on plant growth and development via plant resistance to the fungal pathogen.

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