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1.
Int J Mol Sci ; 21(24)2020 Dec 17.
Artigo em Inglês | MEDLINE | ID: mdl-33348666

RESUMO

Genomic imprinting is an epigenetic phenomenon in which a subset of genes express dependent on the origin of their parents. In plants, it is unclear whether imprinted genes are conserved between subspecies in rice. Here we identified imprinted genes from embryo and endosperm 5-7 days after pollination from three pairs of reciprocal hybrids, including inter-subspecies, japonica intra-subspecies, and indica intra-subspecies reciprocal hybrids. A total of 914 imprinted genes, including 546 in inter-subspecies hybrids, 211 in japonica intra-subspecies hybrids, and 286 in indica intra-subspecies hybrids. In general, the number of maternally expressed genes (MEGs) is more than paternally expressed genes (PEGs). Moreover, imprinted genes tend to be in mini clusters. The number of shared genes by R9N (reciprocal crosses between 9311 and Nipponbare) and R9Z (reciprocal crosses between 9311 and Zhenshan 97), R9N and RZN (reciprocal crosses between Zhonghua11 and Nipponbare), R9Z and RZN was 72, 46, and 16. These genes frequently involved in energy metabolism and seed development. Five imprinted genes (Os01g0151700, Os07g0103100, Os10g0340600, Os11g0679700, and Os12g0632800) are commonly detected in all three pairs of reciprocal hybrids and were validated by RT-PCR sequencing. Gene editing of two imprinted genes revealed that both genes conferred grain filling. Moreover, 15 and 27 imprinted genes with diverse functions in rice were shared with Arabidopsis and maize, respectively. This study provided valuable resources for identification of imprinting genes in rice or even in cereals.


Assuntos
Endosperma/crescimento & desenvolvimento , Endosperma/genética , Metabolismo Energético/genética , Genes de Plantas , Impressão Genômica , Oryza/genética , Oryza/metabolismo , Alelos , Arabidopsis/genética , DNA de Plantas/genética , Epigenômica/métodos , Regulação da Expressão Gênica de Plantas , Família Multigênica , Oryza/embriologia , Polimorfismo de Nucleotídeo Único , RNA de Plantas/genética , Transcriptoma , Zea mays/genética
2.
Int J Mol Sci ; 21(4)2020 Feb 18.
Artigo em Inglês | MEDLINE | ID: mdl-32085665

RESUMO

Absence of high-molecular-weight glutenin subunit (HMW-GS) Dx2 weakens the gluten quality, but it is unclear how the absence of Dx2 has these effects. Thus, we investigated the gluten quality in terms of cytological, physicochemical, and transcriptional characteristics using two near-isogenic lines with Dx2 absent or present at Glu-D1 locus. Cytological observations showed that absence of Dx2 delayed and decreased the accumulation of protein bodies (PBs), where fewer and smaller PBs formed in the endosperm. The activity and gene expression levels of nitrogen assimilation and proteolysis enzymes were lower in HMW-D1a without Dx2 than HMW-D1p with Dx2, and thus less amino acid was transported for protein synthesis in the grains. The expression pattern of genes encoding Glu-1Dx2+1Dy12 was similar to those of three transcription factors, where these genes were significantly down-regulated in HMW-D1a than HMW-D1p. Three genes involving with glutenin polymerization were also down-regulated in HMW-D1a. These results may explain the changes in the glutenin and glutenin macropolymer (GMP) levels during grain development. Therefore, we suggest that the lower nitrogen metabolism capacity and expression levels of glutenin synthesis-related genes in HMW-D1a accounted for the lower accumulation of glutenin, GMP, and PBs, thereby weakening the structural‒thermal properties of gluten.


Assuntos
Regulação da Expressão Gênica de Plantas , Genes de Plantas , Loci Gênicos , Glutens/genética , Nitrogênio/metabolismo , Triticum/genética , Aminoácidos/metabolismo , Endosperma/crescimento & desenvolvimento , Endosperma/metabolismo , Glutamato-Amônia Ligase/metabolismo , Glutens/biossíntese , Glutens/química , Glutens/metabolismo , Folhas de Planta/metabolismo , Polimerização , Subunidades Proteicas/genética , Subunidades Proteicas/metabolismo , Temperatura , Transaminases/metabolismo , Transcrição Genética
3.
BMC Plant Biol ; 20(1): 48, 2020 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-32000683

RESUMO

BACKGROUND: Little is known about the molecular basis of seed size formation in endospermic seed of dicotyledons. The seed of castor bean (Ricinus communis L.) is considered as a model system in seed biology studies because of its persistent endosperms throughout seed development. RESULTS: We compared the size of endosperm and endospermic cells between ZB107 and ZB306 and found that the larger seed size of ZB107 resulted from a higher cell count in the endosperm, which occupy a significant amount of the total seed volume. In addition, fresh weight, dry weight, and protein content of seeds were remarkably higher in ZB107 than in ZB306. Comparative proteomic and transcriptomic analyses were performed between large-seed ZB107 and small-seed ZB306, using isobaric tags for relative and absolute quantification (iTRAQ) and RNA-seq technologies, respectively. A total of 1416 protein species were identified, of which 173 were determined as differentially abundant protein species (DAPs). Additionally, there were 9545 differentially expressed genes (DEGs) between ZB306 and ZB107. Functional analyses revealed that these DAPs and DEGs were mainly involved in cell division and the metabolism of carbohydrates and proteins. CONCLUSIONS: These findings suggest that both cell number and storage-component accumulation are critical for the formation of seed size, providing new insight into the potential mechanisms behind seed size formation in endospermic seeds.


Assuntos
Semente de Rícino/crescimento & desenvolvimento , Proteínas de Plantas/metabolismo , Proteoma/metabolismo , Sementes/crescimento & desenvolvimento , Transcriptoma , Semente de Rícino/genética , Semente de Rícino/metabolismo , Endosperma/genética , Endosperma/crescimento & desenvolvimento , Endosperma/metabolismo , Perfilação da Expressão Gênica , Proteômica , Sementes/genética , Sementes/metabolismo
4.
Plant Sci ; 291: 110336, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31928684

RESUMO

Wheat grain nitrogen content displays large variations within different pearling fractions of grains because of radial gradients in the protein content. We identified how spatiotemporal mechanisms regulate this. The protein gradients emerged clearly at 19 days after anthesis, with the highest N content in aleurone and seed coat, followed by outer endosperm, whereas the lowest was in middle and inner endosperm. Laser microdissection, qRT-PCR and LC-MS were used to dissect tissue from aleurone, outer endosperm, middle endosperm, inner endosperm and transfer cells, measure gene expression and levels of free and protein-bound amino acids, respectively. The results showed that different FAA transportation pathways worked in parallel during grain filling stage while the grain protein gradient did not follow spatial expression of storage proteins. Additionally, two nitrogen (N) topdressing timings were conducted, either at the emergence of top third leaf (standard timing) or top first leaf (delayed timing), finding that delayed N topdressing enhanced both amino acids supply and protein synthesis capacity. The results provide insight into protein synthesis and amino acid transport pathways in endosperm and suggest targets for the enhancement of specialty pearled wheat with higher quality.


Assuntos
Aminoácidos/metabolismo , Endosperma/química , Proteínas de Plantas/metabolismo , Sementes/química , Triticum/genética , Endosperma/crescimento & desenvolvimento , Endosperma/metabolismo , Triticum/química , Triticum/metabolismo
5.
Biochem Biophys Res Commun ; 523(1): 220-225, 2020 02 26.
Artigo em Inglês | MEDLINE | ID: mdl-31848048

RESUMO

Pentatricopeptide repeat (PPR) proteins are one of the largest protein families in land plants. PPR proteins exhibit sequence-specific RNA-binding activity and are implicated in plant growth and development related processes. In this study, we report that the radicleless 1 (rl1) mutant in rice (Oryza sativa L.) exhibited defective radicle emergence in embryos and compromised grain filling in endosperms. Gene cloning and confirmation via genetic complementation analyses showed that RL1 encodes a P-type PPR protein, which is localized to mitochondria. The RL1 protein was specifically involved in the splicing of intron 1 of the mitochondrial nad4 transcript, which encodes a subunit of the mitochondrial NADH dehydrogenase complex. Consistent with this observation, the rl1 mutant exhibited altered mitochondrial morphology and lower ATP accumulation compared with the wild type. Thus, our findings suggest that RL1-mediated nad4 splicing is crucial for embryo and endosperm development in rice.


Assuntos
Endosperma/crescimento & desenvolvimento , Íntrons/genética , Oryza/crescimento & desenvolvimento , Oryza/genética , Proteínas de Plantas/genética , Processamento de RNA/genética , RNA de Plantas/genética , Endosperma/genética , Regulação da Expressão Gênica de Plantas/genética , Oryza/embriologia
6.
Physiol Plant ; 169(1): 110-121, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-31808953

RESUMO

The suspensor is a short-lived tissue critical for proper embryonic development in many higher plants. While the tissue was initially thought to simply suspend the embryo in the endosperm, it has been found through decades of research that it serves multiple important purposes. The suspensor has been found to be vital for proper embryo patterning and numerous studies have been undertaken into the complex transcriptional cross-talk between the suspensor and the embryo proper. Indeed, many suspensor mutants also display abnormalities in the embryo. The suspensor's role as a nutrient conduit has been shown using ultrastructural and histochemical techniques. Biochemical approaches have found that the suspensor is a centre of early embryonic hormone production in several species. The suspensor has also been frequently used as a model for programmed cell death as it shows signs of termination almost immediately upon developing. This review covers the essential functions of the suspensor throughout its short existence from multiple disciplines including structural, genetic and biochemical perspectives.


Assuntos
Apoptose , Endosperma/crescimento & desenvolvimento , Desenvolvimento Vegetal , Sementes/crescimento & desenvolvimento , Plantas
7.
Sci Rep ; 9(1): 18544, 2019 12 06.
Artigo em Inglês | MEDLINE | ID: mdl-31811157

RESUMO

Autophagy plays crucial roles in the recycling of metabolites, and is involved in many developmental processes. Rice mutants defective in autophagy are male sterile due to immature pollens, indicating its critical role in pollen development. However, physiological roles of autophagy during seed maturation had remained unknown. We here found that seeds of the rice autophagy-deficient mutant Osatg7-1, that produces seeds at a very low frequency in paddy fields, are smaller and show chalky appearance and lower starch content in the endosperm at the mature stage under normal growth condition. We comprehensively analyzed the effects of disruption of autophagy on biochemical properties, proteome and seed quality, and found an abnormal activation of starch degradation pathways including accumulation of α-amylases in the endosperm during seed maturation in Osatg7-1. These results indicate critical involvement of autophagy in metabolic regulation in the endosperm of rice, and provide insights into novel autophagy-mediated regulation of starch metabolism during seed maturation.


Assuntos
Proteínas Relacionadas à Autofagia/genética , Autofagia/fisiologia , Endosperma/crescimento & desenvolvimento , Oryza/crescimento & desenvolvimento , Proteínas de Plantas/genética , Proteínas Relacionadas à Autofagia/metabolismo , Endosperma/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Mutação , Oryza/genética , Oryza/metabolismo , Proteínas de Plantas/metabolismo , Amido/metabolismo , Regulação para Cima , alfa-Amilases/genética , alfa-Amilases/metabolismo
8.
Sci Rep ; 9(1): 18567, 2019 12 06.
Artigo em Inglês | MEDLINE | ID: mdl-31811177

RESUMO

In the recent time, Submergence1 (Sub1)QTL, responsible for imparting tolerance to flash flooding, has been introduced in many rice cultivars, but resilience of the QTL to stagnant flooding (SF) is not known. The response of Sub1-introgression has been tested on physiology, molecular biology and yield of two popular rice cultivars (Swarna and Savitri) by comparison of the parental and Sub1-introgression lines (SwarnaSub1 and SavitriSub1) under SF. Compared to control condition SF reduced grain yield and tiller number and increased plant height and Sub1- introgression mostly matched these effects. SF increased ethylene production by over-expression of ACC-synthase and ACC-oxidase enzyme genes of panicle before anthesis in the parental lines. Expression of the genes changed with Sub1-introgression, where some enzyme isoform genes over-expressed after anthesis under SF. Activities of endosperm starch synthesizing enzymes SUS and AGPase declined concomitantly with rise ethylene production in the Sub1-introgressed lines resulting in low starch synthesis and accumulation of soluble carbohydrates in the developing spikelets. In conclusion, Sub1-introgression into the cultivars increased susceptibility to SF. Subjected to SF, the QTL promoted genesis of ethylene in the panicle at anthesis to the detriment of grain yield, while compromising with morphological features like tiller production and stem elongation.


Assuntos
Etilenos/biossíntese , Genes de Plantas/genética , Introgressão Genética , Oryza/fisiologia , Locos de Características Quantitativas , Adaptação Fisiológica/genética , Grão Comestível/crescimento & desenvolvimento , Grão Comestível/metabolismo , Endosperma/crescimento & desenvolvimento , Endosperma/metabolismo , Inundações , Regulação da Expressão Gênica de Plantas , Reguladores de Crescimento de Planta/biossíntese , Brotos de Planta/crescimento & desenvolvimento , Brotos de Planta/metabolismo , Plantas Geneticamente Modificadas/fisiologia , Amido/biossíntese
9.
Plant Sci ; 287: 110203, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31481208

RESUMO

Embryo and endosperm originate from the double fertilization, but they have different developmental fates and biological functions. We identified a previously undescribed maize seed mutant, wherein the embryo appears to be more severely affected than the endosperm (embryo-specific, emb). In the W22 background, the emb embryo arrests at the transition stage whereas its endosperm appears nearly normal in size. At maturity, the embryo in W22-emb is apparently small or even invisible. In contrast, the emb endosperm develops into a relative normal size. We cloned the mutant gene on the Chromosome 7L and designated it emb-7L. This gene is generally expressed, but it has a relatively higher expression level in leaves. Emb-7L encodes a chloroplast-localized P-type pentatricopeptide repeat (PPR) protein, consistent with the severe chloroplast deficiency in emb-7L albino seedling leaves. Full transcriptome analysis of the leaves of WT and emb-7L seedlings reveals that transcription of chloroplast protein-encoding genes are dramatically variable with pre-mRNA intron splicing apparently affected in a tissue-dependent pattern and the chloroplast structure and activity were dramatically affected including chloroplast membrane and photosynthesis machinery component and synthesis of metabolic products (e.g., fatty acids, amino acids, starch).


Assuntos
Proteínas de Plantas/genética , Processamento de RNA , Transcriptoma , Zea mays/genética , Cloroplastos/genética , Cloroplastos/ultraestrutura , Endosperma/embriologia , Endosperma/genética , Endosperma/crescimento & desenvolvimento , Endosperma/ultraestrutura , Regulação da Expressão Gênica de Plantas , Genes de Cloroplastos/genética , Íntrons/genética , Mutação , Fotossíntese , Folhas de Planta/embriologia , Folhas de Planta/genética , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/ultraestrutura , Precursores de RNA/genética , Plântula/embriologia , Plântula/genética , Plântula/crescimento & desenvolvimento , Plântula/ultraestrutura , Zea mays/embriologia , Zea mays/crescimento & desenvolvimento , Zea mays/ultraestrutura
10.
PLoS Genet ; 15(8): e1008305, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31374076

RESUMO

C-to-U editing is an important event in post-transcriptional RNA processing, which converts a specific cytidine (C)-to-uridine (U) in transcripts of mitochondria and plastids. Typically, the pentatricopeptide repeat (PPR) protein, which specifies the target C residue by binding to its upstream sequence, is involved in the editing of one or a few sites. Here we report a novel PPR-DYW protein EMP21 that is associated with editing of 81 sites in maize. EMP21 is localized in mitochondria and loss of the EMP21 function severely inhibits the embryogenesis and endosperm development in maize. From a scan of 35 mitochondrial transcripts produced by the Emp21 loss-of-function mutant, the C-to-U editing was found to be abolished at five sites (nad7-77, atp1-1292, atp8-437, nad3-275 and rps4-870), while reduced at 76 sites in 21 transcripts. In most cases, the failure to editing resulted in the translation of an incorrect residue. In consequence, the mutant became deficient with respect to the assembly and activity of mitochondrial complexes I and V. As six of the decreased editing sites in emp21 overlap with the affected editing sites in emp5-1, and the editing efficiency at rpl16-458 showed a substantial reduction in the emp21-1 emp5-4 double mutant compared with the emp21-1 and emp5-4 single mutants, we explored their interaction. A yeast two hybrid assay suggested that EMP21 does not interact with EMP5, but both EMP21 and EMP5 interact with ZmMORF8. Together, these results indicate that EMP21 is a novel PPR-DYW protein required for the editing of ~17% of mitochondrial target Cs, and the editing process may involve an interaction between EMP21 and ZmMORF8 (and probably other proteins).


Assuntos
Proteínas de Plantas/metabolismo , Edição de RNA , RNA Mitocondrial/metabolismo , Proteínas de Ligação a RNA/metabolismo , Zea mays/fisiologia , Complexo I de Transporte de Elétrons/metabolismo , Desenvolvimento Embrionário/genética , Endosperma/crescimento & desenvolvimento , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Mutação com Perda de Função , Mitocôndrias/metabolismo , Proteínas Mitocondriais/genética , Proteínas Mitocondriais/metabolismo , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Domínios Proteicos/genética , Proteínas de Ligação a RNA/genética
11.
Food Chem ; 301: 125258, 2019 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-31374534

RESUMO

Starch is the abundant component in rice endosperm, and its microstructure determines the quality and functional properties of rice grain. It is well known that the starch fine structure is markedly influenced by high temperature during grain filling. However, it is poorly understood on the competition among starch synthesis related enzymes as well as the interaction between amylose and amylopectin biosynthesis under increased growing temperature. In this study, the non-waxy and waxy rice were planted under normal and high temperatures. Parameterizing analysis of the starch microstructure using mathematical models proved that amylose synthesis competed with the elongation of long amylopectin chains (DP>60); Short chains of amylopectin can be used as the substrate for elongation of longer amylopectin chains; High temperature eliminated the consistency and regularity of the synthesis of amylose and amylopectin. In addition, enzyme assay proved the validity of fitting results from mathematical modeling analysis of starch.


Assuntos
Amilopectina/biossíntese , Amilose/biossíntese , Endosperma/crescimento & desenvolvimento , Endosperma/metabolismo , Temperatura Alta , Oryza/crescimento & desenvolvimento , Oryza/metabolismo
12.
Sci Rep ; 9(1): 11539, 2019 08 08.
Artigo em Inglês | MEDLINE | ID: mdl-31395961

RESUMO

Cyamopsis tetragonoloba (L) endosperm predominantly contains guar gum a polysaccharide, which has tremendous industrial applications in food, textile, paper, oil drilling and water treatment. In order to understand the genes controlling galactomannan biosynthesis, mRNA was isolated from seeds collected at different developmental stages; young pods, mature pods and young leaf from two guar varieties, HG365 and HG870 and subjected to Illumina sequencing. De novo assembly of fourteen individual read files from two varieties of guar representing seven developmental stages gave a total of 1,13,607 contigs with an N50 of 1,244 bases. Annotation of assemblies with GO mapping revealed three levels of distribution, namely, Biological Processes, Molecular Functions and Cellular Components. GO studies identified major genes involved in galactomannan biosynthesis: Cellulose synthase D1 (CS D1) and GAUT-like gene families. Among the polysaccharide biosynthetic process (GO:0000271) genes the transcript abundance for CS was found to be predominantly more in leaf samples, whereas, the transcript abundance for GAUT-like steadily increased from 65% to 90% and above from stage1 to stage5 indicating accumulation of galactomannan in developing seeds; and validated by qRT-PCR analysis. Galactomannan quantification by HPLC showed HG365 (12.98-20.66%) and HG870 (7.035-41.2%) gradually increasing from stage1 to stage 5 (10-50 DAA) and highest accumulation occurred in mature and dry seeds with 3.8 to 7.1 fold increase, respectively. This is the first report of transcriptome sequencing and complete profiling of guar seeds at different developmental stages, young pods, mature pods and young leaf material from two commercially important Indian varieties and elucidation of galactomannan biosynthesis pathway. It is envisaged that the data presented herein will be very useful for improvement of guar through biotechnological interventions in future.


Assuntos
Cyamopsis/genética , Mananas/biossíntese , Desenvolvimento Vegetal/genética , Transcriptoma/genética , Vias Biossintéticas/genética , Metabolismo dos Carboidratos/genética , Cyamopsis/crescimento & desenvolvimento , Endosperma/genética , Endosperma/crescimento & desenvolvimento , Galactanos/genética , Regulação da Expressão Gênica de Plantas , Mananas/genética , Gomas Vegetais/genética , Folhas de Planta/genética , Folhas de Planta/crescimento & desenvolvimento , Sementes/genética , Sementes/crescimento & desenvolvimento , Sequenciamento Completo do Exoma
13.
BMC Genomics ; 20(1): 574, 2019 Jul 11.
Artigo em Inglês | MEDLINE | ID: mdl-31296166

RESUMO

BACKGROUND: Starch biosynthesis in endosperm is a key process influencing grain yield and quality in maize. Although a number of starch biosynthetic genes have been well characterized, the mechanisms by which the expression of these genes is regulated, especially in regard to microRNAs (miRNAs), remain largely unclear. RESULTS: Sequence data for small RNAs, degradome, and transcriptome of maize endosperm at 15 and 25 d after pollination (DAP) from inbred lines Mo17 and Ji419, which exhibit distinct starch content and starch granule structure, revealed the mediation of starch biosynthetic pathways by miRNAs. Transcriptome analysis of these two lines indicated that 33 of 40 starch biosynthetic genes were differentially expressed, of which 12 were up-regulated in Ji419 at 15 DAP, one was up-regulated in Ji419 at 25 DAP, 14 were up-regulated in Ji419 at both 15 and 25 DAP, one was down-regulated in Ji419 at 15 DAP, two were down-regulated in Ji419 at 25 DAP, and three were up-regulated in Ji419 at 15 DAP and down-regulated in Ji419 at 25 DAP, compared with Mo17. Through combined analyses of small RNA and degradome sequences, 22 differentially expressed miRNAs were identified, including 14 known and eight previously unknown miRNAs that could target 35 genes. Furthermore, a complex co-expression regulatory network was constructed, in which 19 miRNAs could modulate starch biosynthesis in endosperm by tuning the expression of 19 target genes. Moreover, the potential operation of four miRNA-mediated pathways involving transcription factors, miR169a-NF-YA1-GBSSI/SSIIIa and miR169o-GATA9-SSIIIa/SBEIIb, was validated via analyses of expression pattern, transient transformation assays, and transactivation assays. CONCLUSION: Our results suggest that miRNAs play a critical role in starch biosynthesis in endosperm, and that miRNA-mediated networks could modulate starch biosynthesis in this tissue. These results have provided important insights into the molecular mechanism of starch biosynthesis in developing maize endosperm.


Assuntos
Perfilação da Expressão Gênica , MicroRNAs/genética , Amido/biossíntese , Zea mays/genética , Zea mays/metabolismo , Endosperma/genética , Endosperma/crescimento & desenvolvimento , Endosperma/metabolismo , Genes de Plantas/genética , Zea mays/crescimento & desenvolvimento
14.
J Appl Genet ; 60(3-4): 233-241, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31297694

RESUMO

Internal reference genes that are stably expressed are essential for normalization in comparative expression analyses. However, gene expression varies significantly among species, organisms, tissues, developmental stages, stresses, and treatments. Therefore, identification of stably expressed reference genes in developmental endosperm of bread wheat is important for expression analysis of endosperm genes. As the first study to systematically screen for reference genes across different developmental stages of wheat endosperm, nine genes were selected from among 76 relatively stable genes based on high-throughput RNA sequencing data. The expression stability of these candidate genes and five traditional reference genes was assessed by real-time quantitative PCR combined with three independent algorithms: geNorm, NormFinder, and BestKeeper. The results showed that ATG8d was the most stable gene during wheat endosperm development, followed by Ta54227, while the housekeeping gene GAPDH, commonly used as an internal reference, was the least stable. ATG8d and Ta54227 together formed the optimal combination of reference genes. Comparative expression analysis of glutenin genes indicated that credible quantification could be achieved by normalization against ATG8d in developmental endosperm. The stably expressed gene characterized here can act as a proper internal reference for expression analysis of wheat endosperm genes, especially nutrient- and nutrient synthesis-related genes.


Assuntos
Endosperma/genética , Genoma de Planta/genética , Desenvolvimento Vegetal/genética , Triticum/genética , Algoritmos , Endosperma/crescimento & desenvolvimento , Regulação da Expressão Gênica no Desenvolvimento/genética , Regulação da Expressão Gênica de Plantas/genética , Padrões de Referência , Triticum/crescimento & desenvolvimento
15.
BMC Plant Biol ; 19(1): 304, 2019 Jul 10.
Artigo em Inglês | MEDLINE | ID: mdl-31291882

RESUMO

BACKGROUND: In flowering plants, proper seed development is achieved through the constant interplay of fertilization products, embryo and endosperm, and maternal tissues. Communication between these compartments is supposed to be tightly regulated at their interfaces. Here, we characterize the deposition pattern of an apoplastic lipid barrier between the maternal inner integument and fertilization products in Arabidopsis thaliana seeds. RESULTS: We demonstrate that an apoplastic lipid barrier is first deposited by the ovule inner integument and undergoes de novo cutin deposition following central cell fertilization and relief of the FERTILIZATION INDEPENDENT SEED Polycomb group repressive mechanism. In addition, we show that the WIP zinc-finger TRANSPARENT TESTA 1 and the MADS-Box TRANSPARENT TESTA 16 transcription factors act maternally to promote its deposition by regulating cuticle biosynthetic pathways. Finally, mutant analyses indicate that this apoplastic barrier allows correct embryo sliding along the seed coat. CONCLUSIONS: Our results revealed that the deposition of a cutin apoplastic barrier between seed maternal and zygotic tissues is part of the seed coat developmental program.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/crescimento & desenvolvimento , Lipídeos de Membrana/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Endosperma/genética , Endosperma/crescimento & desenvolvimento , Endosperma/metabolismo , Óvulo Vegetal/genética , Óvulo Vegetal/crescimento & desenvolvimento , Óvulo Vegetal/metabolismo , Sementes/genética , Sementes/crescimento & desenvolvimento , Sementes/metabolismo
16.
Plant Physiol ; 180(4): 2133-2141, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31221732

RESUMO

Soybean (Glycine max) seed is primarily composed of a mature embryo that provides a major source of protein and oil for humans and other animals. Early in development, the tiny embryos grow rapidly and acquire large quantities of sugars from the liquid endosperm of developing seeds. An insufficient supply of nutrients from the endosperm to the embryo results in severe seed abortion and yield reduction. Hence, an understanding of the molecular basis and regulation of assimilate partitioning involved in early embryo development is important for improving soybean seed yield and quality. Here, we used expression profiling analysis to show that two paralogous sugar transporter genes from the SWEET (Sugars Will Eventually be Exported Transporter) family, GmSWEET15a and GmSWEET15b, were highly expressed in developing soybean seeds. In situ hybridization and quantitative real-time PCR showed that both genes were mainly expressed in the endosperm at the cotyledon stage. GmSWEET15b showed both efflux and influx activities for sucrose in Xenopus oocytes. In Arabidopsis (Arabidopsis thaliana), knockout of three AtSWEET alleles is required to see a defective, but not lethal, embryo phenotype, whereas knockout of both GmSWEET15 genes in soybean caused retarded embryo development and endosperm persistence, resulting in severe seed abortion. In addition, the embryo sugar content of the soybean knockout mutants was greatly reduced. These results demonstrate that the plasma membrane sugar transporter, GmSWEET15, is essential for embryo development in soybean by mediating Suc export from the endosperm to the embryo early in seed development.


Assuntos
Proteínas de Plantas/metabolismo , Sementes/crescimento & desenvolvimento , Sementes/metabolismo , Soja/crescimento & desenvolvimento , Soja/metabolismo , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Endosperma/genética , Endosperma/crescimento & desenvolvimento , Endosperma/metabolismo , Regulação da Expressão Gênica no Desenvolvimento/genética , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Proteínas de Plantas/genética , Reação em Cadeia da Polimerase em Tempo Real , Sementes/genética , Soja/genética
17.
Plant Cell Physiol ; 60(10): 2193-2205, 2019 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-31198964

RESUMO

Tudor-SN is involved in a myriad of transcriptional and post-transcriptional processes due to its modular structure consisting of 4 tandem SN domains (4SN module) and C-terminal Tsn module consisting of Tudor-partial SN domains. We had previously demonstrated that OsTudor-SN is a key player for transporting storage protein mRNAs to specific ER subdomains in developing rice endosperm. Here, we provide genetic evidence that this multifunctional RBP is required for storage protein expression, seed development and protein body formation. The rice EM1084 line, possessing a nonsynonymous mutation in the 4SN module (SN3 domain), exhibited a strong reduction in grain weight and storage protein accumulation, while a mutation in the Tudor domain (47M) or the loss of the Tsn module (43M) had much smaller effects. Immunoelectron microscopic analysis showed the presence of a new protein body type containing glutelin and prolamine inclusions in EM1084, while 43M and 47M exhibited structurally modified prolamine and glutelin protein bodies. Transcriptome analysis indicates that OsTudor-SN also functions in regulating gene expression of transcriptional factors and genes involved in developmental processes and stress responses as well as for storage proteins. Normal protein body formation, grain weight and expression of many genes were partially restored in EM1084 transgenic line complemented with wild-type OsTudor-SN gene. Overall, our study showed that OsTudor-SN possesses multiple functional properties in rice storage protein expression and seed development and that the 4SN and Tsn modules have unique roles in these processes.


Assuntos
Regulação da Expressão Gênica de Plantas , Oryza/genética , Proteínas de Ligação a RNA/metabolismo , Proteínas de Armazenamento de Sementes/metabolismo , Endosperma/genética , Endosperma/crescimento & desenvolvimento , Endosperma/fisiologia , Perfilação da Expressão Gênica , Glutens/metabolismo , Corpos de Inclusão/metabolismo , Mutação , Oryza/crescimento & desenvolvimento , Oryza/fisiologia , Fenilpropanolamina/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Domínios Proteicos , Transporte de RNA , RNA de Plantas/metabolismo , Proteínas de Ligação a RNA/genética , Proteínas de Armazenamento de Sementes/genética
18.
PLoS One ; 14(5): e0217212, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31120929

RESUMO

The endosperm cell wall affects post-harvest grain quality by affecting the mechanical fragility and water absorption of the grain. Therefore, understanding the mechanism underlying endosperm cell wall synthesis is important for determining the growth and quality of cereals. However, the molecular machinery mediating endosperm cell wall biosynthesis is not well understood. In this study, we investigated the role of Oryza sativa Brittle Culm 1-like 6 (OsBC1L6), a member of the COBRA-like protein family, in cellulose synthesis in rice. OsBC1L6 mRNA was expressed in ripening seeds during endosperm enlargement. When OsBC1L6-RFP was expressed in Arabidopsis cell cultures, this fusion protein was transported to the plasma membrane. To investigate the target molecules of OsBC1L6, we analyzed the binding interactions of OsBC1L6 with cellohexaose and the analogs using surface plasmon resonance, determining that cellohexaose bound to OsBC1L6. The ß-glucan contents were significantly reduced in OsBC1L6-RNAi calli and OsBC1L6-deficient seeds from a Tos insertion mutant, compared to their wild-type counterparts. These findings suggest that OsBC1L6 modulates ß-glucan synthesis during endosperm cell wall formation by interacting with cellulose moieties on the plasma membrane during seed ripening.


Assuntos
Parede Celular/metabolismo , Endosperma/metabolismo , Regulação da Expressão Gênica de Plantas , Oryza/metabolismo , Proteínas de Plantas/metabolismo , Sementes/metabolismo , beta-Glucanas/metabolismo , Endosperma/genética , Endosperma/crescimento & desenvolvimento , Oryza/genética , Oryza/crescimento & desenvolvimento , Proteínas de Plantas/genética , Sementes/genética , Sementes/crescimento & desenvolvimento
19.
Plant Cell Rep ; 38(9): 1099-1107, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31127322

RESUMO

KEY MESSAGE: MdoDHN11 acts in the nucellus layer to protect the embryo and the endosperm from limited water availability during apple seed development. Dehydrins (DHNs) are protective proteins related to several plant developmental responses that involve dehydration such as seed desiccation and abiotic stresses. In apple (Malus × domestica Borkh.), the seed-specific MdoDHN11 was suggested to play important roles against dehydration during seed development. However, this hypothesis has not yet been evaluated. Within this context, several experiments were performed to functionally characterize MdoDHN11. In situ hybridization analysis during apple seed development showed that MdoDHN11 expression is confined to a maternal tissue called nucellus, a central mass of parenchyma between the endosperm and the testa. The MdoDHN11 protein was localized in the cytosol and nucleus. Finally, transgenic Arabidopsis plants expressing MdoDHN11 were generated and exposed to a severe water-deficit stress, aiming to mimic a situation that can occurs during seed development. All transgenic lines showed increased tolerance to water deficit in relation to wild-type plants. Taken together, our results provide evidences that MdoDHN11 plays important roles during apple seed development by protecting the embryo and the endosperm from limited water availability, and the mechanism of action probably involves the interaction of MdoDHN11 with proteins and other components in the cell.


Assuntos
Malus/genética , Proteínas de Plantas/metabolismo , Água/fisiologia , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/fisiologia , Desidratação , Endosperma/genética , Endosperma/crescimento & desenvolvimento , Endosperma/fisiologia , Expressão Gênica , Malus/crescimento & desenvolvimento , Malus/fisiologia , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Sementes/genética , Sementes/crescimento & desenvolvimento , Sementes/fisiologia
20.
J Exp Bot ; 70(18): 4705-4720, 2019 09 24.
Artigo em Inglês | MEDLINE | ID: mdl-31087099

RESUMO

Pentatricopeptide repeat (PPR) proteins constitute one of the largest protein families in land plants. Recent studies revealed the functions of PPR proteins in organellar RNA metabolism and plant development, but the functions of most PPR proteins, especially PPRs localized in the nucleus, remain largely unknown. Here, we report the isolation and characterization of a rice mutant named floury and growth retardation1 (fgr1). fgr1 showed floury endosperm with loosely arranged starch grains, decreased starch and amylose contents, and retarded seedling growth. Map-based cloning showed that the mutant phenotype was caused by a single nucleotide substitution in the coding region of Os08g0290000. This gene encodes a nuclear-localized PPR protein, which we named OsNPPR1, that affected mitochondrial function. In vitro SELEX and RNA-EMSAs showed that OsNPPR1 was an RNA protein that bound to the CUCAC motif. Moreover, a number of retained intron (RI) events were detected in fgr1. Thus, OsNPPR1 was involved in regulation of mitochondrial development and/or functions that are important for endosperm development. Our results provide novel insights into coordinated interaction between nuclear-localized PPR proteins and mitochondrial function.


Assuntos
Endosperma/crescimento & desenvolvimento , Mitocôndrias/metabolismo , Oryza/genética , Proteínas de Plantas/genética , Endosperma/metabolismo , Oryza/metabolismo , Proteínas de Plantas/metabolismo
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