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1.
Curr Protoc ; 1(7): e191, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34242495

RESUMO

Well-characterized genetic resources are fundamental to maintain and provide the various genotypes for pre-breeding programs for the production of new cultivars (e.g., wild relatives, unimproved material, landraces). The aim of the current article is to provide protocols for the characterization of the genetic resources of two lupin crop species: the European Lupinus albus and the American Lupinus mutabilis. Intelligent nested collections of lupins derived from homozygous lines (single-seed descent) are being developed, established, and exploited using cutting-edge approaches for genotyping, phenotyping, data management, and data analysis within the INCREASE project (EU Horizon 2020). This will allow us to predict the phenotypic performance of genotyped lines, and will further boost research and development in lupins. Lupins stand out due to their high-quality seed protein (∼40% of seed dry weight) and other primary components in the seeds, which include fatty acids, dietary fiber, and minerals. The potential of lupins as a crop is highlighted by the multiple benefits of plant-based food in terms of food security, nutrition, human health, and sustainable production. The use of lupins in foods, along with other well-studied and widely used food legumes, will also provide a greatly diversified plant-based food palette to meet the Global Goals for Sustainable Development to improve people's lives by 2030. © 2021 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Lupin seed phenotypic descriptors Basic Protocol 2: Lupin seed imaging Basic Protocol 3: Standardized phenotypic characterization of lupin genetic resources grown towards primary seed increase (development of single-seed descent genetic resources).


Assuntos
Lupinus , Fibras na Dieta , Genótipo , Humanos , Lupinus/genética , Melhoramento Vegetal , Sementes/genética
2.
Int J Mol Sci ; 22(13)2021 Jun 26.
Artigo em Inglês | MEDLINE | ID: mdl-34206810

RESUMO

Recently, crop breeders have widely adopted a new biotechnology-based process, termed Seed Production Technology (SPT), to produce hybrid varieties. The SPT does not produce nuclear male-sterile lines, and instead utilizes transgenic SPT maintainer lines to pollinate male-sterile plants for propagation of nuclear-recessive male-sterile lines. A late-stage pollen-specific promoter is an essential component of the pollen-inactivating cassette used by the SPT maintainers. While a number of plant pollen-specific promoters have been reported so far, their usefulness in SPT has remained limited. To increase the repertoire of pollen-specific promoters for the maize community, we conducted a comprehensive comparative analysis of transcriptome profiles of mature pollen and mature anthers against other tissue types. We found that maize pollen has much less expressed genes (>1 FPKM) than other tissue types, but the pollen grain has a large set of distinct genes, called pollen-specific genes, which are exclusively or much higher (100 folds) expressed in pollen than other tissue types. Utilizing transcript abundance and correlation coefficient analysis, 1215 mature pollen-specific (MPS) genes and 1009 mature anther-specific (MAS) genes were identified in B73 transcriptome. These two gene sets had similar GO term and KEGG pathway enrichment patterns, indicating that their members share similar functions in the maize reproductive process. Of the genes, 623 were shared between the two sets, called mature anther- and pollen-specific (MAPS) genes, which represent the late-stage pollen-specific genes of the maize genome. Functional annotation analysis of MAPS showed that 447 MAPS genes (71.7% of MAPS) belonged to genes encoding pollen allergen protein. Their 2-kb promoters were analyzed for cis-element enrichment and six well-known pollen-specific cis-elements (AGAAA, TCCACCA, TGTGGTT, [TA]AAAG, AAATGA, and TTTCT) were found highly enriched in the promoters of MAPS. Interestingly, JA-responsive cis-element GCC box (GCCGCC) and ABA-responsive cis-element-coupling element1 (ABRE-CE1, CCACC) were also found enriched in the MAPS promoters, indicating that JA and ABA signaling likely regulate pollen-specific MAPS expression. This study describes a robust and straightforward pipeline to discover pollen-specific promotes from publicly available data while providing maize breeders and the maize industry a number of late-stage (mature) pollen-specific promoters for use in SPT for hybrid breeding and seed production.


Assuntos
Perfilação da Expressão Gênica/métodos , Melhoramento Vegetal/métodos , Pólen/genética , Transcriptoma , Zea mays/genética , Regulação da Expressão Gênica de Plantas , Infertilidade das Plantas/genética , Pólen/metabolismo , Regiões Promotoras Genéticas , Sementes/genética , Sementes/metabolismo , Zea mays/metabolismo
3.
Planta ; 254(2): 32, 2021 Jul 21.
Artigo em Inglês | MEDLINE | ID: mdl-34287699

RESUMO

MAIN CONCLUSION: A ß-ketoacyl-ACP-synthase II (KAS2) like enzyme and a lysophosphatidic acid acyltransferase (LPAT2) from Consolida ajacis catalyze gondoic acid biosynthesis and incorporation into the sn-2 position of seed TAG in engineered Camelina sativa. Gondoic acid (cis-11 eicosenoic acid, 20:1∆11) is the predominant very-long-chain fatty acid (VLCFA) in camelina (Camelina sativa) seed oil accounting for 12-15% of total triacylglycerol fatty acids. To explore the feasibility of engineering increased levels of this fatty acid in camelina seed, oils from a range of plant species were analyzed to identify those producing 20-Carbon (C20) fatty acids as the only VLCFAs in their seed oil. Seeds of Consolida and Delphinium species (Ranunculaceae) were found to contain moderate levels (0.2% to 25.5%) of C20 fatty acids without accompanying longer chain fatty acids. The C20 fatty acids were abundant in both sn-2 and sn-1/3 positions of seed TAG in Consolida, but were largely absent from the sn-2 position in Delphinium seed TAG. Through generation of a developing seed transcriptome, sequences were identified and cDNAs amplified from Consolida ajacis encoding a ß-ketoacyl-ACP-synthase II like protein (CaKAS2B) that lacked a predicted chloroplast transit peptide, and two homologues of Arabidopsis thaliana lysophosphatidic acid acyltransferase 2 (CaLPAT2a and CaLPAT2b). Expression of CaKAS2B in conventional (WT) camelina and a line previously engineered for high seed oleic acid content (HO) resulted in increased seed VLCFA content. Total VLCFA levels were raised from 24 to 35% and from 7 to 23% in T3 seed from representative transformants in the WT and HO backgrounds, respectively. Gondoic acid was the predominant VLCFA in transformed HO lines with low endogenous cytoplasmic fatty acid elongation activity, suggesting limited capacity of CaKAS2B to elongate beyond C20. Expression in camelina of CaLPAT2b resulted in significantly increased C20-VLCFA esterification at the sn-2 position of seed TAG with VLCFA levels of 33.8% in this position in one transformed line compared to 0.3% at sn-2 in the corresponding control line. Only small changes in total seed VLCFA content were observed in transformed lines implying that increased VLCFA esterification capacity in camelina results in positional redistribution of VLCFAs but does not significantly enhance flux through the fatty acid elongation pathway. The full potential of CaKAS2B and CaLPAT2a for the engineering of high gondoic acid levels in camelina remains to be determined. Seed fatty acid composition of Consolida and Delphinium also provides information that may be of value in the systematics of the Ranunculaceae.


Assuntos
Brassicaceae , Delphinium , Brassicaceae/genética , DNA Complementar/genética , Expressão Ectópica do Gene , Ácidos Graxos , Ácidos Graxos Monoinsaturados , Óleos Vegetais , Plantas Geneticamente Modificadas , Sementes/genética , Triglicerídeos
4.
Planta ; 254(2): 37, 2021 Jul 26.
Artigo em Inglês | MEDLINE | ID: mdl-34309737

RESUMO

MAIN CONCLUSION: AS-3 line of Sorghum bicolor possesses functional components of apomixis-apospory, parthenogenesis and autonomous endospermogenesis. The data obtained indicate efficiency of selection for apomixis components in diploid species of cultivated crops. Apomixis (seed formation without fertilization) is one of most attractive phenomena in plant biology. In this paper, we provide the results of long-term selection for apomixis components in the progeny of grain sorghum (Sorghum bicolor (L.) Moench) hybrid plants with male sterility mutation. Selection was carried out for a high frequency of aposporous embryo sacs (ESs), autonomous pro-embryos, and the presence of maternal-type plants in test crosses with the line Volzhskoe-4v (V4v) homozygous for the Rs1 genes determining the red color of the leaves and stem of the hybrids. As a result of using this approach, the line, AS-3, was created, in which the frequency of ovaries with parthenogenetic embryos reached 42-45%. The autonomous development of embryos and endosperm was observed in the panicles of each of the 10 cytologically studied plants of this line. The frequency of parthenogenesis positively correlated with the high average daily air temperature during the first five out of 10 days preceding the onset of flowering (r = 0.75; P > 0.01). Genotyping of the plants from the progeny of hand-emasculated panicles of AS-3 pollinated with V4v performed using co-dominant SSR markers revealed that the F1 hybrids carrying the Rs1 gene (chromosome 6) possessed both paternal and maternal alleles of Sb1-10 (chromosome 4) and Xtxp320 (chromosome 10) markers, while in the maternal-type plants (rs1rs1), only the maternal alleles of these markers were present. In the endosperm of the kernels from which the maternal-type seedlings were obtained, only the maternal alleles were present, while in the endosperm of the kernels that produced hybrid seedlings, both the paternal and maternal alleles were observed. The data obtained indicate the presence of functional components of apomixis (apospory, parthenogenesis, autonomous endospermogenesis) in the grain sorghum line AS-3, and the efficiency of selection for apomixis in functionally diploid species of cultivated crops.


Assuntos
Apomixia , Sorghum , Diploide , Endosperma/genética , Sementes/genética , Sorghum/genética
5.
Int J Mol Sci ; 22(11)2021 Jun 04.
Artigo em Inglês | MEDLINE | ID: mdl-34199720

RESUMO

The underground reserve (root) has been an uncharted research territory with its untapped genetic variation yet to be exploited. Identifying ideal traits and breeding new rice varieties with efficient root system architecture (RSA) has great potential to increase resource-use efficiency and grain yield, especially under direct-seeded rice, by adapting to aerobic soil conditions. In this review, we tried to mine the available research information on the direct-seeded rice (DSR) root system to highlight the requirements of different root traits such as root architecture, length, number, density, thickness, diameter, and angle that play a pivotal role in determining the uptake of nutrients and moisture at different stages of plant growth. RSA also faces several stresses, due to excess or deficiency of moisture and nutrients, low or high temperature, or saline conditions. To counteract these hindrances, adaptation in response to stress becomes essential. Candidate genes such as early root growth enhancer PSTOL1, surface rooting QTL qSOR1, deep rooting gene DRO1, and numerous transporters for their respective nutrients and stress-responsive factors have been identified and validated under different circumstances. Identifying the desired QTLs and transporters underlying these traits and then designing an ideal root architecture can help in developing a suitable DSR cultivar and aid in further advancement in this direction.


Assuntos
Adaptação Fisiológica/fisiologia , Oryza/fisiologia , Raízes de Plantas/anatomia & histologia , Raízes de Plantas/fisiologia , Sementes/fisiologia , Germinação/fisiologia , Oryza/genética , Oryza/crescimento & desenvolvimento , Locos de Características Quantitativas/genética , Sementes/genética , Sementes/crescimento & desenvolvimento
6.
BMC Plant Biol ; 21(1): 311, 2021 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-34210282

RESUMO

BACKGROUND: Mineral elements are important for maintaining good human health besides heavy metals. Mining genes that control mineral elements are paramount for improving their accumulation in the wheat grain. Although previous studies have reported some loci for beneficial trace elements, they have mainly focused on Zn and Fe content. However, little information is available regarding the genetic loci differences in dissecting synchronous accumulation of multiple mineral elements in wheat grains, including beneficial and heavy elements. Therefore, a genome-wide association study (GWAS) was conducted on 205 wheat accessions with 24,355 single nucleotide polymorphisms (SNPs) to identify important loci and candidate genes for controlling Ca, Fe, Zn, Se, Cu, Mn, Cd, As, and Pb accumulation in wheat grains. RESULTS: A total of 101 marker-trait associations (MTAs) (P < 10-5) loci affecting the content of nine mineral elements was identified on chromosomes 1B, 1D, 2A, 2B, 3A, 3B, 3D, 4A, 4B, 5A, 5B, 5D, 6B, 7A, 7B, and 7D. Among these, 17 major MTAs loci for the nine mineral elements were located, and four MTAs loci (P < 10-5) were found on chromosomes 1B, 6B, 7B, and 7D. Eight multi-effect MTAs loci were detected that are responsible for the control of more than one trait, mainly distributed on chromosomes 3B, 7B, and 5A. Furthermore, sixteen candidate genes controlling Ca, Fe, Zn, Se, Cd, and Pb were predicted, whose functions were primarily related to ion binding, including metals, Fe, Ca, Cu, Mg, and Zn, ATP binding, ATPase activity, DNA binding, RNA binding, and protein kinase activity. CONCLUSIONS: Our study indicated the existence of gene interactions among mineral elements based on multi-effect MTAs loci and candidate genes. Meanwhile this study provided new insights into the genetic control of mineral element concentrations, and the important loci and genes identified may contribute to the rapid development of beneficial mineral elements and a reduced content of harmful heavy metals in wheat grain.


Assuntos
Genoma de Planta , Minerais/metabolismo , Estações do Ano , Sementes/genética , Triticum/genética , Alelos , Mapeamento Cromossômico , Loci Gênicos , Marcadores Genéticos , Estudo de Associação Genômica Ampla , Fenótipo
7.
BMC Plant Biol ; 21(1): 313, 2021 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-34215178

RESUMO

BACKGROUND: Harnessing heterosis is one of the major approaches to increase rice yield and has made a great contribution to food security. The identification and selection of outstanding parental genotypes especially among male sterile lines is a key step for exploiting heterosis. Two-line hybrid system is based on the discovery and application of photoperiod- and thermo-sensitive genic sensitive male sterile (PTGMS) materials. The development of wide-range of male sterile lines from a common gene pool leads to a narrower genetic diversity, which is vulnerable to biotic and abiotic stress. Hence, it is valuable to ascertain the genetic background of PTGMS lines and to understand their relationships in order to select and design a future breeding strategy. RESULTS: A collection of 118 male sterile rice lines and 13 conventional breeding lines from the major rice growing regions of China was evaluated and screened against the photosensitive (pms3) and temperature sensitive male sterility (tms5) genes. The total gene pool was divided into four major populations as P1 possessing the pms3, P2 possessing tms5, P3 possessing both pms3 and tms5 genes, and P4 containing conventional breeding lines without any male sterility allele. The high genetic purity was revealed by homozygous alleles in all populations. The population admixture, principle components and the phylogenetic analysis revealed the close relations of P2 and P3 with P4. The population differentiation analysis showed that P1 has the highest differentiation coefficient. The lines from P1 were observed as the ancestors of other three populations in a phylogenetic tree, while the lines in P2 and P3 showed a close genetic relation with conventional lines. A core collection of top 10% lines with maximum within and among populations genetic diversity was constructed for future research and breeding efforts. CONCLUSION: The low genetic diversity and close genetic relationship among PTGMS lines in P2, P3 and P4 populations suggest a selection sweep and they might result from a backcrossing with common ancestors including the pure lines of P1. The core collection from PTGMS panel updated with new diverse germplasm will serve best for further two-line hybrid breeding.


Assuntos
Oryza/genética , Fotoperíodo , Infertilidade das Plantas/genética , Sementes/genética , Temperatura , Núcleo Celular/genética , Núcleo Celular/efeitos da radiação , Análise por Conglomerados , Ontologia Genética , Estudos de Associação Genética , Marcadores Genéticos , Luz , Nucleotídeos/genética , Oryza/efeitos da radiação , Filogenia , Infertilidade das Plantas/efeitos da radiação , Polimorfismo de Nucleotídeo Único/genética , Análise de Componente Principal , Reprodutibilidade dos Testes , Sementes/efeitos da radiação
8.
BMC Plant Biol ; 21(1): 330, 2021 Jul 09.
Artigo em Inglês | MEDLINE | ID: mdl-34243721

RESUMO

BACKGROUND: The climate crisis threatens sustainability of crop production worldwide. Crop diversification may enhance food security while reducing the negative impacts of climate change. Proso millet (Panicum milaceum L.) is a minor cereal crop which holds potential for diversification and adaptation to different environmental conditions. In this study, we assembled a world collection of proso millet consisting of 88 varieties and landraces to investigate its genomic and phenotypic diversity for seed traits, and to identify marker-trait associations (MTA). RESULTS: Sequencing of restriction-site associated DNA fragments yielded 494 million reads and 2,412 high quality single nucleotide polymorphisms (SNPs). SNPs were used to study the diversity in the collection and perform a genome wide association study (GWAS). A genotypic diversity analysis separated accessions originating in Western Europe, Eastern Asia and Americas from accessions sampled in Southern Asia, Western Asia, and Africa. A Bayesian structure analysis reported four cryptic genetic groups, showing that landraces accessions had a significant level of admixture and that most of the improved proso millet materials clustered separately from landraces. The collection was highly diverse for seed traits, with color varying from white to dark brown and width spanning from 1.8 to 2.6 mm. A GWAS study for seed morphology traits identified 10 MTAs. In addition, we identified three MTAs for agronomic traits that were previously measured on the collection. CONCLUSION: Using genomics and automated seed phenotyping, we elucidated phylogenetic relationships and seed diversity in a global millet collection. Overall, we identified 13 MTAs for key agronomic and seed traits indicating the presence of alleles with potential for application in proso breeding programs.


Assuntos
Biodiversidade , Produtos Agrícolas/genética , Grão Comestível/genética , Estudo de Associação Genômica Ampla , Panicum/genética , Fenótipo , Sementes/genética , Europa (Continente) , Genótipo , Filogenia , Polimorfismo de Nucleotídeo Único
9.
Int J Mol Sci ; 22(13)2021 Jul 05.
Artigo em Inglês | MEDLINE | ID: mdl-34281271

RESUMO

The allotetraploid species Brassica juncea (mustard) is grown worldwide as oilseed and vegetable crops; the yellow seed-color trait is particularly important for oilseed crops. Here, to examine the factors affecting seed coat color, we performed a metabolic and transcriptomic analysis of yellow- and dark-seeded B. juncea seeds. In this study, we identified 236 compounds, including 31 phenolic acids, 47 flavonoids, 17 glucosinolates, 38 lipids, 69 other hydroxycinnamic acid compounds, and 34 novel unknown compounds. Of these, 36 compounds (especially epicatechin and its derivatives) accumulated significantly different levels during the development of yellow- and dark-seeded B. juncea. In addition, the transcript levels of BjuDFR, BjuANS,BjuBAN, BjuTT8, and BjuTT19 were closely associated with changes to epicatechin and its derivatives during seed development, implicating this pathway in the seed coat color determinant in B. juncea. Furthermore, we found numerous variations of sequences in the TT8A genes that may be associated with the stability of seed coat color in B. rapa, B. napus, and B. juncea, which might have undergone functional differentiation during polyploidization in the Brassica species. The results provide valuable information for understanding the accumulation of metabolites in the seed coat color of B. juncea and lay a foundation for exploring the underlying mechanism.


Assuntos
Mostardeira/genética , Mostardeira/metabolismo , Catequina/análogos & derivados , Catequina/metabolismo , Flavonoides/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Glucosinolatos/metabolismo , Metaboloma , Mostardeira/crescimento & desenvolvimento , Fenótipo , Pigmentação/genética , Sementes/genética , Sementes/metabolismo
10.
Int J Mol Sci ; 22(13)2021 Jun 29.
Artigo em Inglês | MEDLINE | ID: mdl-34209672

RESUMO

Gene editing by use of clustered regularly interspaced short palindromic repeats (CRISPR) has become a powerful tool for crop improvement. However, a common bottleneck in the application of this approach to grain crops, including rice (Oryza sativa), is efficient vector delivery and calli regeneration, which can be hampered by genotype-dependent requirements for plant regeneration. Here, methods for Agrobacterium-mediated and biolistic transformation and regeneration of indica rice were optimized using CRISPR-Cas9 gene-editing of the submergence tolerance regulator SUBMERGENCE 1A-1 gene of the cultivar Ciherang-Sub1. Callus induction and plantlet regeneration methods were optimized for embryogenic calli derived from immature embryos and mature seed-derived calli. Optimized regeneration (95%) and maximal editing efficiency (100%) were obtained from the immature embryo-derived calli. Phenotyping of T1 seeds derived from the edited T0 plants under submergence stress demonstrated inferior phenotype compared to their controls, which phenotypically validates the disruption of SUB1A-1 function. The methods pave the way for rapid CRISPR-Cas9 gene editing of recalcitrant indica rice cultivars.


Assuntos
Genes de Plantas , Oryza/fisiologia , Regeneração , Transformação Genética , Sistemas CRISPR-Cas , Proteínas de Ligação ao Cálcio/genética , Edição de Genes , Fenótipo , Desenvolvimento Vegetal/genética , Plantas Geneticamente Modificadas , Reprodutibilidade dos Testes , Sementes/genética , Sementes/crescimento & desenvolvimento
11.
Int J Mol Sci ; 22(13)2021 Jun 29.
Artigo em Inglês | MEDLINE | ID: mdl-34209706

RESUMO

The plant transcription factor WRINKLED1 (WRI1), a member of AP2/EREBP, is involved in the regulation of glycolysis and the expression of genes related to the de novo synthesis of fatty acids in plastids. In this study, the key regulator of seed oil synthesis and accumulation transcription factor gene PoWRI1 was identified and cloned, having a complete open reading frame of 1269 bp and encoding 422 amino acids. Subcellular localization analysis showed that PoWRI1 is located at the nucleus. After the expression vector of PoWRI1 was constructed and transformed into wild-type Arabidopsis thaliana, it was found that the overexpression of PoWRI1 increased the expression level of downstream target genes such as BCCP2, KAS1, and PKP-ß1. As a result, the seeds of transgenic plants became larger, the oil content increased significantly, and the unsaturated fatty acid content increased, which provide a scientific theoretical basis for the subsequent use of genetic engineering methods to improve the fatty acid composition and content of plant seeds.


Assuntos
Regulação da Expressão Gênica de Plantas , Paeonia/genética , Paeonia/metabolismo , Óleos Vegetais/metabolismo , Proteínas de Plantas/genética , Arabidopsis/genética , Arabidopsis/metabolismo , Sequência de Bases , Vias Biossintéticas/genética , Clonagem Molecular , Ácidos Graxos/metabolismo , Fenótipo , Filogenia , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Transporte Proteico , Sementes/genética , Sementes/metabolismo , Análise de Sequência de DNA
12.
Nat Commun ; 12(1): 3963, 2021 06 25.
Artigo em Inglês | MEDLINE | ID: mdl-34172749

RESUMO

The endosperm provides nutrients and growth regulators to the embryo during seed development. LEAFY COTYLEDON1 (LEC1) has long been known to be essential for embryo maturation. LEC1 is expressed in both the embryo and the endosperm; however, the functional relevance of the endosperm-expressed LEC1 for seed development is unclear. Here, we provide genetic and transgenic evidence demonstrating that endosperm-expressed LEC1 is necessary and sufficient for embryo maturation. We show that endosperm-synthesized LEC1 is capable of orchestrating full seed maturation in the absence of embryo-expressed LEC1. Inversely, without LEC1 expression in the endosperm, embryo development arrests even in the presence of functional LEC1 alleles in the embryo. We further reveal that LEC1 expression in the endosperm begins at the zygote stage and the LEC1 protein is then trafficked to the embryo to activate processes of seed maturation. Our findings thus establish a key role for endosperm in regulating embryo development.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/crescimento & desenvolvimento , Proteínas Estimuladoras de Ligação a CCAAT/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Proteínas Estimuladoras de Ligação a CCAAT/genética , Endosperma/genética , Endosperma/crescimento & desenvolvimento , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Haploidia , Plantas Geneticamente Modificadas , Sementes/genética , Sementes/crescimento & desenvolvimento
13.
BMC Plant Biol ; 21(1): 270, 2021 Jun 11.
Artigo em Inglês | MEDLINE | ID: mdl-34116632

RESUMO

BACKGROUND: Cinnamomum migao is an endangered evergreen woody plant species endemic to China. Its fruit is used as a traditional medicine by the Miao nationality of China and has a high commercial value. However, its seed germination rate is extremely low under natural and artificial conditions. As the foundation of plant propagation, seed germination involves a series of physiological, cellular, and molecular changes; however, the molecular events and systematic changes occurring during C. migao seed germination remain unclear. RESULTS: In this study, combined with the changes in physiological indexes and transcription levels, we revealed the regulation characteristics of cell structures, storage substances, and antioxidant capacity during seed germination. Electron microscopy analysis revealed that abundant smooth and full oil bodies were present in the cotyledons of the seeds. With seed germination, oil bodies and other substances gradually degraded to supply energy; this was consistent with the content of storage substances. In parallel to electron microscopy and physiological analyses, transcriptome analysis showed that 80-90 % of differentially expressed genes (DEGs) appeared after seed imbibition, reflecting important development and physiological changes. The unigenes involved in material metabolism (glycerolipid metabolism, fatty acid degradation, and starch and sucrose metabolism) and energy supply pathways (pentose phosphate pathway, glycolysis pathway, pyruvate metabolism, tricarboxylic acid cycle, and oxidative phosphorylation) were differentially expressed in the four germination stages. Among these DEGs, a small number of genes in the energy supply pathway at the initial stage of germination maintained high level of expression to maintain seed vigor and germination ability. Genes involved in lipid metabolism were firstly activated at a large scale in the LK (seed coat fissure) stage, and then genes involved in carbohydrates (CHO) metabolism were activated, which had their own species specificity. CONCLUSIONS: Our study revealed the transcriptional levels of genes and the sequence of their corresponding metabolic pathways during seed germination. The changes in cell structure and physiological indexes also confirmed these events. Our findings provide a foundation for determining the molecular mechanisms underlying seed germination.


Assuntos
Cinnamomum/genética , Cinnamomum/metabolismo , Perfilação da Expressão Gênica , Germinação/genética , Germinação/fisiologia , Plantas Medicinais/genética , Plantas Medicinais/fisiologia , China , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Sementes/genética , Sementes/metabolismo
14.
Braz J Biol ; 83: e246455, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34161463

RESUMO

The inheritance of the seedless fruit characteristic of Annona squamosa has not yet been explained. Molecular techniques may aid breeding programs, mainly in the assisted selection of the target gene. The INO gene may be related to seed development in these fruits. The objective of the present paper was to investigate the inheritance of seedlessness in the 'Brazilian seedless' sugar apple and INO gene conservation in Annona squamosa and Annona cherimola x Annona squamosa genotypes by assessing their homology with the INO database genes. The F1 generation was obtained by crossing the mutant 'Brazilian seedless' (male genitor) (P1) with the wild-type A. squamosa with seeds (M1 and M2, female genitors). The INO gene was studied in mutant and wild-type A. squamosa (P1, M1, M2 and M3) and in the Gefner atemoya (A. cherimola x A. squamosa) (M4) cultivar. The DNA was extracted from young leaves, and four sets of specific primers flanking the INO gene were amplified. The seedless characteristic was identified as stenospermatic in the fruits of parental P1, suggesting monogenic inheritance with complete dominance. High sequence similarity of the INO gene amplifications in the sugar apple accessions (M1, M2, M3) and the atemoya cultivar Gefner (M4) reinforces the hypothesis of their conservation.


Assuntos
Annona , Annonaceae , Annona/genética , Brasil , Frutas/genética , Melhoramento Vegetal , Sementes/genética
15.
Gene ; 793: 145749, 2021 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-34077776

RESUMO

Maize ZmGS5 was reported to be positively associated with kernel-related traits, however, its regulatory mechanism on plant development and seed size remains unknown. In this study, ZmGS5 was demonstrated to be widely expressed in various maize tissues with the highest expression level in developing embryos, indicating its critical roles in early kernel development process. The ZmGS5 protein was subcellularly localized to both the nucleus and cytoplasm. Transgenic Arabidopsis plants overexpressing ZmGS5 under the control of either the constitutive maize Ubiquitin1 promotor or native ZmGS5 promoter resulted in increased plant size, biomass, seed size and weight, although no significant difference was observed between transgenic lines harboring the two constructs. In contrast, the antisense-ZmGS5 transgene resulted in opposite phenotypes. Our cytological data suggested that ZmGS5 enlarged petal size through enhancing cell expansion. Quantitative RT-PCR analysis indicated that ZmGS5 might enhance cell expansion and grain filling by upregulating expression levels of particular EXPA or SWEET genes. Collectively, these findings help us further understand the biological function and regulatory mechanism of ZmGS5 in improving organ size and seed weight, which imply its great potential for high-yield breeding in the future.


Assuntos
Arabidopsis/genética , Regulação da Expressão Gênica de Plantas , Sementes/genética , Transgenes , Zea mays/genética , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/metabolismo , Contagem de Células , Núcleo Celular/metabolismo , Núcleo Celular/ultraestrutura , Citoplasma/metabolismo , Citoplasma/ultraestrutura , Humanos , Tamanho do Órgão , Melhoramento Vegetal/métodos , Células Vegetais/metabolismo , Plantas Geneticamente Modificadas , Regiões Promotoras Genéticas , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Sementes/anatomia & histologia , Sementes/crescimento & desenvolvimento , Sementes/metabolismo , Ubiquitina/genética , Ubiquitina/metabolismo , Zea mays/crescimento & desenvolvimento , Zea mays/metabolismo
16.
BMC Genomics ; 22(1): 453, 2021 Jun 16.
Artigo em Inglês | MEDLINE | ID: mdl-34134624

RESUMO

BACKGROUND: Seeds are the economic basis of oilseed crops, especially soybeans, the most widely cultivated oilseed crop worldwide. Seed development is accompanied by a multitude of diverse cellular processes, and revealing the underlying regulatory activities is critical for seed improvement. RESULTS: In this study, we profiled the transcriptomes of developing seeds at 20, 25, 30, and 40 days after flowering (DAF), as these stages represent critical time points of seed development from early to full development. We identified a set of highly abundant genes and highlighted the importance of these genes in supporting nutrient accumulation and transcriptional regulation for seed development. We identified 8925 differentially expressed genes (DEGs) that exhibited temporal expression patterns over the course and expression specificities in distinct tissues, including seeds and nonseed tissues (roots, stems, and leaves). Genes specific to nonseed tissues might have tissue-associated roles, with relatively low transcript abundance in developing seeds, suggesting their spatially supportive roles in seed development. Coexpression network analysis identified several underexplored genes in soybeans that bridge tissue-specific gene modules. CONCLUSIONS: Our study provides a global view of gene activities and biological processes critical for seed formation in soybeans and prioritizes a set of genes for further study. The results of this study help to elucidate the mechanism controlling seed development and storage reserves.


Assuntos
Regulação da Expressão Gênica de Plantas , Soja , Perfilação da Expressão Gênica , Sementes/genética , Soja/genética , Transcriptoma
17.
J Food Sci ; 86(7): 2962-2977, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34076269

RESUMO

Alcalase hydrolyzates were prepared from the albumin (AH) and globulin (GH) fractions of eight chickpea (Cicer arietinum L.) genotypes from Mexico and 10 from other countries. Protein content, antioxidant activity (AA) (ABTS, DPPH), and degree of hydrolysis were evaluated and the best genotype was selected by principal component analysis. The hydrolyzates of the chosen genotype were analyzed for its antidiabetic potential measured as inhibition of α-amylase, α-glucosidase, and dipeptidyl peptidase-4 (DPP4). Peptide profiles were obtained by liquid chromatography-mass spectrometry (UPLC-DAD-MS), and the most active peptides were analyzed by molecular docking. The average antioxidant activity of albumin hydrolyzates was higher than that of globulin hydrolyzates. ICC3761 was the selected genotype and peptides purified from the albumin hydrolyzate showed the best antioxidant activity and antidiabetic potential (FEI, FEL, FIE, FKN, FGKG, and MEE). FEI, FEL, and FIE were in the same chromatographic peak and this mixture showed the best ABTS scavenging (78.25%) and DPP4 inhibition (IC50  = 4.20 µg/ml). MEE showed the best DPPH scavenging (47%). FGKG showed the best inhibition of α-amylase (54%) and α-glucosidase (56%) and may be a competitive inhibitor based on in silico-predicted interactions with catalytic amino acids in the active site of both enzymes. These peptides could be used as nutraceutical supplements against diseases related to oxidative stress and diabetes. PRACTICAL APPLICATION: This study showed that chickpea protein hydrolyzates are good sources of peptides with antidiabetic potential, showing high antioxidant activity and inhibition of enzymes related to carbohydrate metabolism and type 2 diabetes. These hydrolyzates could be formulated in functional foods for diabetes.


Assuntos
Antioxidantes/química , Cicer/química , Hipoglicemiantes/química , Peptídeos/química , Proteínas de Plantas/química , Cromatografia Líquida , Cicer/genética , Dipeptidil Peptidase 4/química , Inibidores da Dipeptidil Peptidase IV/química , Genótipo , Humanos , Espectrometria de Massas , Simulação de Acoplamento Molecular , Extratos Vegetais/química , Hidrolisados de Proteína/química , Sementes/química , Sementes/genética , alfa-Amilases/química , alfa-Glucosidases/química
18.
Plant Sci ; 309: 110913, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34134840

RESUMO

Abscisic acid (ABA) is an important regulator of plant growth, development, and biotic and abiotic stress responses. Ubiquitination plays important roles in regulating ABA signaling. E3 ligase, a key member in ubiquitination, actively participates in the regulation of biosynthesis, de-repression, and activation of ABA response and degradation of signaling components. In this study, we found that that overexpression of wheat E3 ligase TaPUB1 decreased the sensitivity of wheat seedlings to ABA, whereas TaPUB1-RNA interference (TaPUB1-RNAi) lines increased wheat sensitivity to ABA during germination, root growth, and stomatal opening. TaPUB1 influenced the expression of several ABA-responsive genes, and also interacted with TaPYL4 and TaABI5, which are involved in ABA signal transduction, and promoted their degradation. Additionally, we observed that TaPUB1-OE lines resulted in lower single-split grain numbers, larger seed size, and higher thousand kernel weight, when compared with the WT lines. Contrasting results were obtained for TaPUB1-RNAi lines. It suggests that TaPUB1 acts as a negative regulator in the ABA signaling pathway by interacting with TaPYL4 and TaABI5, subsequently affecting seed development in wheat. In addition, the enhanced abiotic tolerance of overexpression lines due to enhanced photosynthesis and root development may be related to the degradation of TaABI5 by TaPUB1.


Assuntos
Ácido Abscísico/metabolismo , Reguladores de Crescimento de Plantas/metabolismo , Transdução de Sinais , Triticum/genética , Germinação , Fotossíntese , Plântula/genética , Plântula/crescimento & desenvolvimento , Plântula/fisiologia , Sementes/genética , Sementes/crescimento & desenvolvimento , Sementes/fisiologia , Estresse Fisiológico , Triticum/crescimento & desenvolvimento , Triticum/fisiologia , Ubiquitina-Proteína Ligases/genética , Ubiquitina-Proteína Ligases/metabolismo , Ubiquitinação
19.
Int J Mol Sci ; 22(10)2021 May 18.
Artigo em Inglês | MEDLINE | ID: mdl-34069840

RESUMO

The biosynthesis and storage of lipids in oil crop seeds involve many gene families, such as nonspecific lipid-transfer proteins (nsLTPs). nsLTPs are cysteine-rich small basic proteins essential for plant development and survival. However, in sesame, information related to nsLTPs was limited. Thus, the objectives of this study were to identify the Sesamum indicum nsLTPs (SiLTPs) and reveal their potential role in oil accumulation in sesame seeds. Genome-wide analysis revealed 52 SiLTPs, nonrandomly distributed on 10 chromosomes in the sesame variety Zhongzhi 13. Following recent classification methods, the SiLTPs were divided into nine types, among which types I and XI were the dominants. We found that the SiLTPs could interact with several transcription factors, including APETALA2 (AP2), DNA binding with one finger (Dof), etc. Transcriptome analysis showed a tissue-specific expression of some SiLTP genes. By integrating the SiLTPs expression profiles and the weighted gene co-expression network analysis (WGCNA) results of two contrasting oil content sesame varieties, we identified SiLTPI.23 and SiLTPI.28 as the candidate genes for high oil content in sesame seeds. The presumed functions of the candidate gene were validated through overexpression of SiLTPI.23 in Arabidopsis thaliana. These findings expand our knowledge on nsLTPs in sesame and provide resources for functional studies and genetic improvement of oil content in sesame seeds.


Assuntos
Proteínas de Transferência de Fosfolipídeos/genética , Proteínas de Transferência de Fosfolipídeos/metabolismo , Sesamum/genética , Proteínas de Transporte/metabolismo , Etiquetas de Sequências Expressas , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas/genética , Genes de Plantas/genética , Óleos Vegetais/metabolismo , Sementes/genética , Sesamum/metabolismo , Fatores de Transcrição/metabolismo
20.
Int J Mol Sci ; 22(11)2021 May 29.
Artigo em Inglês | MEDLINE | ID: mdl-34072567

RESUMO

In recent years, much progress has been made in elucidating the functional roles of plant glycine-rich RNA-binding proteins (GR-RBPs) during development and stress responses. Canonical GR-RBPs contain an RNA recognition motif (RRM) or a cold-shock domain (CSD) at the N-terminus and a glycine-rich domain at the C-terminus, which have been associated with several different RNA processes, such as alternative splicing, mRNA export and RNA editing. However, many aspects of GR-RBP function, the targeting of their RNAs, interacting proteins and the consequences of the RNA target process are not well understood. Here, we discuss recent findings in the field, newly defined roles for GR-RBPs and the actions of GR-RBPs on target RNA metabolism.


Assuntos
Regulação da Expressão Gênica de Plantas , Desenvolvimento Vegetal/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Proteínas de Ligação a RNA/genética , Proteínas de Ligação a RNA/metabolismo , Estresse Fisiológico , Germinação/genética , Filogenia , Processamento Pós-Transcricional do RNA , Sementes/genética , Sementes/metabolismo
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