RESUMEN
Teosinte, the wild ancestor of maize (Zea mays subsp. mays), has three times the seed protein content of most modern inbreds and hybrids, but the mechanisms that are responsible for this trait are unknown1,2. Here we use trio binning to create a contiguous haplotype DNA sequence of a teosinte (Zea mays subsp. parviglumis) and, through map-based cloning, identify a major high-protein quantitative trait locus, TEOSINTE HIGH PROTEIN 9 (THP9), on chromosome 9. THP9 encodes an asparagine synthetase 4 enzyme that is highly expressed in teosinte, but not in the B73 inbred, in which a deletion in the tenth intron of THP9-B73 causes incorrect splicing of THP9-B73 transcripts. Transgenic expression of THP9-teosinte in B73 significantly increased the seed protein content. Introgression of THP9-teosinte into modern maize inbreds and hybrids greatly enhanced the accumulation of free amino acids, especially asparagine, throughout the plant, and increased seed protein content without affecting yield. THP9-teosinte seems to increase nitrogen-use efficiency, which is important for promoting a high yield under low-nitrogen conditions.
Asunto(s)
Nitrógeno , Zea mays , Zea mays/genética , Familia , Semillas/genéticaRESUMEN
Development of the cereal endosperm involves cell differentiation processes that enable nutrient uptake from the maternal plant, accumulation of storage products, and their utilization during germination. However, little is known about the regulatory mechanisms that link cell differentiation processes with those controlling storage product synthesis and deposition, including the activation of zein genes by the maize (Zea mays) bZIP transcription factor Opaque-2 (O2). Here, we mapped in vivo binding sites of O2 in B73 endosperm and compared the results with genes differentially expressed in B73 and B73o2 We identified 186 putative direct O2 targets and 1677 indirect targets, encoding a broad set of gene functionalities. Examination of the temporal expression patterns of O2 targets revealed at least two distinct modes of O2-mediated gene activation. Two O2-activated genes, bZIP17 and NAKED ENDOSPERM2 (NKD2), encode transcription factors, which can in turn coactivate other O2 network genes with O2. NKD2 (with its paralog NKD1) was previously shown to be involved in regulation of aleurone development. Collectively, our results provide insights into the complexity of the O2-regulated network and its role in regulation of endosperm cell differentiation and function.
Asunto(s)
Endospermo/citología , Redes Reguladoras de Genes , Proteínas de Plantas/genética , Zea mays/genética , Sitios de Unión , Diferenciación Celular , Inmunoprecipitación de Cromatina , Endospermo/genética , Regulación de la Expresión Génica de las Plantas , Mutación , Células Vegetales/fisiología , Proteínas de Plantas/metabolismo , Zea mays/citologíaRESUMEN
Endosperm is a filial structure resulting from a second fertilization event in angiosperms. As an absorptive storage organ, endosperm plays an essential role in support of embryo development and seedling germination. The accumulation of carbohydrate and protein storage products in cereal endosperm provides humanity with a major portion of its food, feed, and renewable resources. Little is known regarding the regulatory gene networks controlling endosperm proliferation and differentiation. As a first step toward understanding these networks, we profiled all mRNAs in the maize kernel and endosperm at eight successive stages during the first 12 d after pollination. Analysis of these gene sets identified temporal programs of gene expression, including hundreds of transcription-factor genes. We found a close correlation of the sequentially expressed gene sets with distinct cellular and metabolic programs in distinct compartments of the developing endosperm. The results constitute a preliminary atlas of spatiotemporal patterns of endosperm gene expression in support of future efforts for understanding the underlying mechanisms that control seed yield and quality.
Asunto(s)
Endospermo/metabolismo , Regulación del Desarrollo de la Expresión Génica/genética , Regulación de la Expresión Génica de las Plantas/genética , Zea mays/genética , Biología Computacional , Perfilación de la Expresión Génica , Reacción en Cadena en Tiempo Real de la Polimerasa , Análisis de Secuencia de ARN , Factores de Tiempo , Zea mays/metabolismoRESUMEN
In angiosperms, the endosperm provides nutrients for embryogenesis and seed germination and is the primary tissue where gene imprinting occurs. To identify the imprintome of early developing maize (Zea mays) endosperm, we performed high-throughput transcriptome sequencing of whole kernels at 0, 3, and 5 d after pollination (DAP) and endosperms at 7, 10, and 15 DAP, using B73 by Mo17 reciprocal crosses. We observed gradually increased expression of paternal transcripts in 3- and 5-DAP kernels. In 7-DAP endosperm, the majority of the genes tested reached a 2:1 maternal versus paternal ratio, suggesting that paternal genes are nearly fully activated by 7 DAP. A total of 116, 234, and 63 genes exhibiting parent-specific expression were identified at 7, 10, and 15 DAP, respectively. The largest proportion of paternally expressed genes was at 7 DAP, mainly due to the significantly deviated parental allele expression ratio of these genes at this stage, while nearly 80% of the maternally expressed genes (MEGs) were specific to 10 DAP and were primarily attributed to sharply increased expression levels compared with the other stages. Gene ontology enrichment analysis of the imprinted genes suggested that 10-DAP endosperm-specific MEGs are involved in nutrient uptake and allocation and the auxin signaling pathway, coincident with the onset of starch and storage protein accumulation.
Asunto(s)
Endospermo/genética , Regulación de la Expresión Génica de las Plantas/genética , Genoma de Planta/genética , Impresión Genómica/genética , Transcriptoma , Zea mays/genética , Alelos , Secuencia de Bases , Diferenciación Celular , Metilación de ADN , Endospermo/crecimiento & desarrollo , Endospermo/metabolismo , Perfilación de la Expresión Génica , Biblioteca de Genes , Ontología de Genes , Secuenciación de Nucleótidos de Alto Rendimiento , Especificidad de Órganos , Polinización , Polimorfismo de Nucleótido Simple , ARN de Planta/química , ARN de Planta/genética , Semillas/genética , Semillas/crecimiento & desarrollo , Semillas/metabolismo , Análisis de Secuencia de ARN , Almidón/metabolismo , Zea mays/crecimiento & desarrollo , Zea mays/metabolismoRESUMEN
The endosperm of cereal grains is one of the most valuable products of modern agriculture. Cereal endosperm development comprises different phases characterized by mitotic cell proliferation, endoreduplication, the accumulation of storage compounds, and programmed cell death. Although manipulation of these processes could maximize grain yield, how they are regulated and integrated is poorly understood. We show that the Retinoblastoma-related (RBR) pathway controls key aspects of endosperm development in maize. Down-regulation of RBR1 by RNAi resulted in up-regulation of RBR3-type genes, as well as the MINICHROMOSOME MAINTENANCE 2-7 gene family and PROLIFERATING CELL NUCLEAR ANTIGEN, which encode essential DNA replication factors. Both the mitotic and endoreduplication cell cycles were stimulated. Developing transgenic endosperm contained 42-58% more cells and â¼70% more DNA than wild type, whereas there was a reduction in cell and nuclear sizes. In addition, cell death was enhanced. The DNA content of mature endosperm increased 43% upon RBR1 down-regulation, whereas storage protein content and kernel weight were essentially not affected. Down-regulation of both RBR1 and CYCLIN DEPENDENT KINASE A (CDKA);1 indicated that CDKA;1 is epistatic to RBR1 and controls endoreduplication through an RBR1-dependent pathway. However, the repressive activity of RBR1 on downstream targets was independent from CDKA;1, suggesting diversification of RBR1 activities. Furthermore, RBR1 negatively regulated CDK activity, suggesting the presence of a feedback loop. These results indicate that the RBR1 pathway plays a major role in regulation of different processes during maize endosperm development and suggest the presence of tissue/organ-level regulation of endosperm/seed homeostasis.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Endospermo/fisiología , Proteína de Retinoblastoma/metabolismo , Zea mays/metabolismo , Ciclo Celular , Muerte Celular , Proliferación Celular , Quinasas Ciclina-Dependientes/metabolismo , Regulación de la Expresión Génica de las Plantas , Genotipo , Plantas Modificadas Genéticamente/genética , Plantas Modificadas Genéticamente/metabolismo , Interferencia de ARN , Semillas/fisiología , Zea mays/genéticaRESUMEN
Endosperm development in maize (Zea mays L.) and related cereals comprises a cell proliferation stage followed by a period of rapid growth coupled to endoreduplication. Regulation of the cell cycle in developing endosperm is poorly understood. We have characterized various subunits of cyclin-dependent kinase (CDK) complexes, master cell cycle regulators in all eukaryotes. A-, B-, and D-type cyclins as well as A- and B-type cyclin-dependent kinases were characterized with respect to their RNA and protein expression profiles. Two main patterns were identified: one showing expression throughout endosperm development, and another characterized by a sharp down-regulation with the onset of endoreduplication. Cyclin CYCB1;3 and CYCD2;1 proteins were distributed in the cytoplasm and nucleus of cells throughout the endosperm, while cyclin CYCD5 protein was localized in the cytoplasm of peripheral cells. CDKB1;1 expression was strongly associated with cell proliferation. Expression and cyclin-binding patterns suggested that CDKA;1 and CDKA;3 are at least partially redundant. The kinase activity associated with the cyclin CYCA1 was highest during the mitotic stage of development, while that associated with CYCB1;3, CYCD2;1 and CYCD5 peaked at the mitosis-to-endoreduplication transition. A-, B- and D-type cyclins were more resistant to proteasome-dependent degradation in endoreduplicating than in mitotic endosperm extracts. These results indicated that endosperm development is characterized by differential expression and activity of specific cyclins and CDKs, and suggested that endoreduplication is associated with reduced cyclin proteolysis via the ubiquitin-proteasome pathway.
Asunto(s)
Quinasas Ciclina-Dependientes/genética , Regulación de la Expresión Génica de las Plantas , Zea mays/enzimología , Animales , División Celular , Aumento de la Célula , Núcleo Celular/metabolismo , Células Cultivadas , Quinasas Ciclina-Dependientes/metabolismo , Regulación hacia Abajo , Drosophila , Endorreduplicación , Endospermo/enzimología , Endospermo/genética , Mitosis , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Complejo de la Endopetidasa Proteasomal , Proteínas Recombinantes de Fusión , Semillas/enzimología , Semillas/genética , Análisis de Secuencia de ADN , Zea mays/citología , Zea mays/genéticaRESUMEN
UNLABELLED: ⢠PREMISE OF THE STUDY: Given the worldwide economic importance of maize endosperm, it is surprising that its development is not the most comprehensively studied of the cereals. We present detailed morphometric and cytological descriptions of endosperm development in the maize inbred line B73, for which the genome has been sequenced, and compare its growth with four diverse Nested Association Mapping (NAM) founder lines.⢠METHODS: The first 12 d of B73 endosperm development were described using semithin sections of plastic-embedded kernels and confocal microscopy. Longitudinal sections were used to compare endosperm length, thickness, and area.⢠KEY RESULTS: Morphometric comparison between Arizona- and Michigan-grown B73 showed a common pattern. Early endosperm development was divided into four stages: coenocytic, cellularization through alveolation, cellularization through partitioning, and differentiation. We observed tightly synchronous nuclear divisions in the coenocyte, elucidated that the onset of cellularization was coincident with endosperm size, and identified a previously undefined cell type (basal intermediate zone, BIZ). NAM founders with small mature kernels had larger endosperms (0-6 d after pollination) than lines with large mature kernels.⢠CONCLUSIONS: Our B73-specific model of early endosperm growth links developmental events to relative endosperm size, while accounting for diverse growing conditions. Maize endosperm cellularizes through alveolation, then random partitioning of the central vacuole. This unique cellularization feature of maize contrasts with the smaller endosperms of Arabidopsis, barley, and rice that strictly cellularize through repeated alveolation. NAM analysis revealed differences in endosperm size during early development, which potentially relates to differences in timing of cellularization across diverse lines of maize.
Asunto(s)
Diferenciación Celular , Endospermo/crecimiento & desarrollo , Zea mays/crecimiento & desarrollo , Endospermo/anatomía & histología , Endospermo/citología , Fertilización , Células Vegetales , Polinización , Especificidad de la Especie , Zea mays/anatomía & histología , Zea mays/citologíaRESUMEN
Dent and flint kernel architectures are important characteristics that affect the physical properties of maize kernels and their grain end uses. The genes controlling these traits are unknown, so it is difficult to combine the advantageous kernel traits of both. We found mutation of ARFTF17 in a dent genetic background reduces IAA content in the seed pericarp, creating a flint-like kernel phenotype. ARFTF17 is highly expressed in the pericarp and encodes a protein that interacts with and inhibits MYB40, a transcription factor with the dual functions of repressing PIN1 expression and transactivating genes for flavonoid biosynthesis. Enhanced flavonoid biosynthesis could reduce the metabolic flux responsible for auxin biosynthesis. The decreased IAA content of the dent pericarp appears to reduce cell division and expansion, creating a shorter, denser kernel. Introgression of the ARFTF17 mutation into dent inbreds and hybrids improved their kernel texture, integrity, and desiccation, without affecting yield.
Asunto(s)
Semillas , Zea mays , Zea mays/genética , Zea mays/metabolismo , Fenotipo , Semillas/genética , Mutación , Flavonoides/metabolismoRESUMEN
BACKGROUND: The opaque2 mutant is valuable for producing maize varieties with enhanced nutritional value. However, the exact mechanisms by which it improves protein quality and creates a soft endosperm texture are unclear. Given the importance of improving nutritional quality in grain crops, a better understanding of the physiological basis for these traits is necessary. RESULTS: In this study, we combined transcript profiling and proteomic analysis to better understand which genes and proteins are altered by opaque2 in the W64A inbred line. These analyses showed that the accumulation of some lysine-rich proteins, such as sorbitol dehydrogenase and glyceraldehyde3-phosphate dehydrogenase, was increased in mature kernels and may contribute substantially to the lysine content of opaque2 endosperm. Some defense proteins such as beta-glucosidase aggregating factor were strongly down regulated and may be regulated directly by opaque2. The mutant also had altered expression of a number of starch biosynthesis genes and this was associated with a more highly crystalline starch. CONCLUSIONS: The results of these studies revealed specific target genes that can be investigated to further improve nutritional quality and agronomic performance of high lysine maize lines, particularly those based on the presence of the opaque2 mutation. Alteration of amylopectin branching patterns in opaque2 starch could contribute to generation of the soft, starchy endosperm.
Asunto(s)
Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Proteómica , Almidón/biosíntesis , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Transcripción Genética , Zea mays/metabolismo , Proteínas de Unión al ADN/química , Endospermo/genética , Endospermo/crecimiento & desarrollo , Endospermo/metabolismo , Regulación de la Expresión Génica de las Plantas , Lisina/metabolismo , Mutación , Proteínas de Plantas/química , Semillas/genética , Semillas/crecimiento & desarrollo , Semillas/metabolismo , Factores de Transcripción/química , Zea mays/química , Zea mays/genética , Zea mays/crecimiento & desarrolloRESUMEN
Retinoblastoma-related (RBR) genes inhibit the cell cycle primarily by repressing adenovirus E2 promoter binding factor (E2F) transcription factors, which drive the expression of numerous genes required for DNA synthesis and cell cycle progression. The RBR-E2F pathway is conserved in plants, but cereals such as maize are characterized by having a complex RBR gene family with at least 2 functionally distinct members, RBR1 and RBR3. Although RBR1 has a clear cell cycle inhibitory function, it is not known whether RBR3 has a positive or negative role. By uncoupling RBR3 from the negative regulation of RBR1 in cultured maize embryos through a combination of approaches, we demonstrate that RBR3 has a positive and critical role in the expression of E2F targets required for the initiation of DNA synthesis, DNA replication, and the efficiency with which transformed plants can be obtained. Titration of endogenous RBR3 activity through expression of a dominant-negative allele with a compromised pocket domain suggests that these RBR3 functions require an activity distinct from its pocket domain. Our results indicate a cell cycle pathway in maize, in which 2 RBR genes have specific and opposing functions. Thus, the paradigm that RBR genes are negative cell cycle regulators cannot be considered universal.
Asunto(s)
Cromosomas de las Plantas/genética , Replicación del ADN , Regulación de la Expresión Génica de las Plantas , Genes de Retinoblastoma , Proteínas de Plantas/genética , Zea mays/citología , Zea mays/genética , Regulación hacia Abajo/genética , Fase G2 , Genes de Plantas , Modelos Genéticos , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente , ARN Mensajero/genética , ARN Mensajero/metabolismo , Transformación GenéticaRESUMEN
Quality protein maize (QPM) is a high lysine-containing corn that is based on genetic modification of the opaque2 (o2) mutant. In QPM, modifier genes convert the starchy endosperm of o2 to the vitreous phenotype of wild type maize. There are multiple, unlinked o2 modifier loci (Opm) in QPM and their nature and mode of action are unknown. We previously identified seven Opm QTLs and characterized 16 genes that are differentially up-regulated at a significant level in K0326Y QPM, compared to the starchy endosperm mutant W64Ao2. In order to further characterize these Opm QTLs and the genes up-regulated in K0326Y QPM, we created a population of 314 recombinant inbred lines (RILs) from a cross between K0326Y QPM and W64Ao2. The RILs were characterized for three traits associated with endosperm texture: vitreousness, density and hardness. Genetic linkage analysis of the RIL population confirmed three of the previously identified QTLs associated with o2 endosperm modification in K0326Y QPM. Many of the genes up-regulated in K0326Y QPM showed substantially higher levels of expression in vitreous compared with opaque RILs. These included genes associated with the upstream regulation of the ethylene response pathway, and a gene encoding a regulatory subunit of pyrophosphate-dependent fructose-6-phosphate 1-phosphotransferase, an adaptive enzyme of the glycolytic pathway.
Asunto(s)
Proteínas de Unión al ADN/genética , Genes de Plantas/genética , Estudios de Asociación Genética , Endogamia , Proteínas Nucleares/genética , Proteínas de Plantas/genética , Sitios de Carácter Cuantitativo/genética , Recombinación Genética/genética , Factores de Transcripción/genética , Zea mays/genética , Mapeo Cromosómico , Cromosomas de las Plantas/genética , Regulación de la Expresión Génica de las Plantas , Dureza , Peso Molecular , Fenotipo , Carácter Cuantitativo Heredable , Zeína/genética , Zeína/metabolismoRESUMEN
In eukaryotes, alternative splicing of pre-mRNAs contributes significantly to the proper expression of the genome. However, the functions of many auxiliary spliceosomal proteins are still unknown. Here, we functionally characterized plant homologues of nematode suppressors of mec-8 and unc-52 (smu). We compared transcript profiles of maize (Zea mays) smu2 endosperm with those of wild-type plants and identified pre-mRNA splicing events that depend on the maize SMU2 protein. Consistent with a conserved role of plant SMU-2 homologues, Arabidopsis (Arabidopsis thaliana) smu2 mutants also show altered splicing of similar target pre-mRNAs. The Atsmu2 mutants occasionally show developmental phenotypes, including abnormal cotyledon numbers and higher seed weights. We identified AtSMU1 as one of the SMU2-interacting proteins, and Atsmu1 mutations cause similar developmental phenotypes with higher penetrance than Atsmu2. The AtSMU2 and AtSMU1 proteins are localized to the nucleus and highly prevalent in actively dividing tissues. Taken together, our data indicated that the plant SMU-1 and SMU-2 homologues appear to be involved in splicing of specific pre-mRNAs that affect multiple aspects of development.
Asunto(s)
Empalme Alternativo , Arabidopsis/genética , Precursores del ARN/genética , ARN de Planta/genética , Zea mays/genética , Arabidopsis/metabolismo , ADN Bacteriano/genética , Perfilación de la Expresión Génica , Regulación del Desarrollo de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Mutagénesis Insercional , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente/genética , Plantas Modificadas Genéticamente/metabolismo , Mapeo de Interacción de Proteínas , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Zea mays/metabolismoRESUMEN
In plants, the amino acids tyrosine and phenylalanine are synthesized from arogenate by arogenate dehydrogenase and arogenate dehydratase, respectively, with the relative flux to each being tightly controlled. Here the characterization of a maize opaque endosperm mutant (mto140), which also shows retarded vegetative growth, is described The opaque phenotype co-segregates with a Mutator transposon insertion in an arogenate dehydrogenase gene (zmAroDH-1) and this led to the characterization of the four-member family of maize arogenate dehydrogenase genes (zmAroDH-1-zmAroDH-4) which share highly similar sequences. A Mutator insertion at an equivalent position in AroDH-3, the most closely related family member to AroDH-1, is also associated with opaque endosperm and stunted vegetative growth phenotypes. Overlapping but differential expression patterns as well as subtle mutant effects on the accumulation of tyrosine and phenylalanine in endosperm, embryo, and leaf tissues suggest that the functional redundancy of this gene family provides metabolic plasticity for the synthesis of these important amino acids. mto140/arodh-1 seeds shows a general reduction in zein storage protein accumulation and an elevated lysine phenotype typical of other opaque endosperm mutants, but it is distinct because it does not result from quantitative or qualitative defects in the accumulation of specific zeins but rather from a disruption in amino acid biosynthesis.
Asunto(s)
Regulación de la Expresión Génica de las Plantas , Prefenato Deshidrogenasa/genética , Prefenato Deshidrogenasa/metabolismo , Semillas , Zea mays/enzimología , Zea mays/genética , Secuencia de Aminoácidos , Aminoácidos/metabolismo , Endospermo/enzimología , Endospermo/genética , Lisina/metabolismo , Datos de Secuencia Molecular , Mutación/genética , Fenotipo , Fenilalanina/biosíntesis , Fenilalanina/genética , Fenilalanina/metabolismo , Prefenato Deshidrogenasa/química , Semillas/enzimología , Semillas/genética , Semillas/metabolismo , Alineación de Secuencia , Tirosina/metabolismo , Zea mays/clasificación , Zea mays/metabolismo , Zeína/metabolismoRESUMEN
The mechanism that creates vitreous endosperm in the mature maize kernel is poorly understood. We identified Vitreous endosperm 1 (Ven1) as a major QTL influencing this process. Ven1 encodes ß-carotene hydroxylase 3, an enzyme that modulates carotenoid composition in the amyloplast envelope. The A619 inbred contains a nonfunctional Ven1 allele, leading to a decrease in polar and an increase in non-polar carotenoids in the amyloplast. Coincidently, the stability of amyloplast membranes is increased during kernel desiccation. The lipid composition in endosperm cells in A619 is altered, giving rise to a persistent amyloplast envelope. These changes impede the gathering of protein bodies and prevent them from interacting with starch grains, creating air spaces that cause an opaque kernel phenotype. Genetic modifiers were identified that alter the effect of Ven1A619, while maintaining a high ß-carotene level. These studies provide insight for breeding vitreous kernel varieties and high vitamin A content in maize.
Asunto(s)
Carotenoides/metabolismo , Zea mays/metabolismo , Alelos , Mapeo Cromosómico , Cruzamientos Genéticos , Endospermo/genética , Endospermo/metabolismo , Endospermo/ultraestructura , Genes de Plantas , Microscopía Electrónica de Rastreo , Microscopía Electrónica de Transmisión , Oxigenasas de Función Mixta/genética , Oxigenasas de Función Mixta/metabolismo , Fenotipo , Fitomejoramiento , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plastidios/genética , Plastidios/metabolismo , Plastidios/ultraestructura , Sitios de Carácter Cuantitativo , Semillas/genética , Semillas/metabolismo , Semillas/ultraestructura , Zea mays/genética , Zea mays/ultraestructuraRESUMEN
Mutation of o2 doubles maize endosperm lysine content, but it causes an inferior kernel phenotype. Developing quality protein maize (QPM) by introgressing o2 modifiers (Mo2s) into the o2 mutant benefits millions of people in developing countries where maize is a primary protein source. Here, we report genome sequence and annotation of a South African QPM line K0326Y, which is assembled from single-molecule, real-time shotgun sequencing reads collinear with an optical map. We achieve a N50 contig length of 7.7 million bases (Mb) directly from long-read assembly, compared to those of 1.04 Mb for B73 and 1.48 Mb for Mo17. To characterize Mo2s, we map QTLs to chromosomes 1, 6, 7, and 9 using an F2 population derived from crossing K0326Y and W64Ao2. RNA-seq analysis of QPM and o2 endosperms reveals a group of differentially expressed genes that coincide with Mo2 QTLs, suggesting a potential role in vitreous endosperm formation.
Asunto(s)
Proteínas de Plantas/genética , Zea mays/genética , Mapeo Cromosómico , Endospermo/genética , Regulación de la Expresión Génica de las Plantas , Genoma de Planta , Genómica , Sitios de Carácter CuantitativoRESUMEN
Development of the seed endosperm involves several different types of coordinated cell cycle programs: acytokinetic mitosis, which produces a syncytium soon after fertilization; cellularization through the formation of modified phragmoplasts; cell proliferation, in which mitosis is coupled to cell division; and, in certain species like cereal crops, endoreduplication. Understanding the regulation of these programs and their transitions is challenging, but it has the potential to define important links between the cell cycle, cell differentiation and development, as well as provide tools for the manipulation of seed yield. A relatively large number of mutants display endosperm proliferation defects, and connections with known cell cycle genes are beginning to emerge. For example, it is becoming increasingly evident that the master cell cycle regulators, the cyclin-dependent kinases and retinoblastoma-related families, play key roles in the events leading to endosperm formation and development. Recent studies highlight cross-talk between pathways controlling the cell cycle and genomic imprinting.
Asunto(s)
Arabidopsis/citología , Ciclo Celular , Endospermo/embriología , Zea mays/citología , Arabidopsis/embriología , Arabidopsis/genética , Arabidopsis/metabolismo , Endospermo/citología , Endospermo/genética , Endospermo/metabolismo , Regulación de la Expresión Génica de las Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Zea mays/embriología , Zea mays/genética , Zea mays/metabolismoRESUMEN
Since its development more than two decades ago, Quality Protein Maize (QPM) has been adopted for cultivation in many regions of the developing world. Given the potential benefits of widespread use of QPM, research to better understand the genetic and biochemical mechanisms responsible for its altered kernel texture and protein quality is important. Recent investigations into the improved protein quality of the opaque2 mutant and the genetic mechanisms that can suppress its starchy kernel phenotype provide new insights to support the continued improvement of QPM. Chief among these developments are the use of transgenic approaches to improve nutritional quality and the discovery that an important component of modified endosperm texture in QPM is related to altered starch granule structure.
Asunto(s)
Cromosomas de las Plantas/genética , Proteínas de Unión al ADN/genética , Proteínas de Plantas/genética , Plantas Modificadas Genéticamente/genética , Factores de Transcripción/genética , Zea mays/genética , Mutación/genética , Almidón/química , Almidón/genéticaRESUMEN
BACKGROUND: Gene knockouts are a critical resource for functional genomics. In Arabidopsis, comprehensive knockout collections were generated by amplifying and sequencing genomic DNA flanking insertion mutants. These Flanking Sequence Tags (FSTs) map each mutant to a specific locus within the genome. In maize, FSTs have been generated using DNA transposons. Transposable elements can generate unstable insertions that are difficult to analyze for simple knockout phenotypes. Transposons can also generate somatic insertions that fail to segregate in subsequent generations. RESULTS: Transposon insertion sites from 106 UniformMu FSTs were tested for inheritance by locus-specific PCR. We confirmed 89% of the FSTs to be germinal transposon insertions. We found no evidence for somatic insertions within the 11% of insertion sites that were not confirmed. Instead, this subset of insertion sites had errors in locus-specific primer design due to incomplete or low-quality genomic sequences. The locus-specific PCR assays identified a knockout of a 6-phosphogluconate dehydrogenase gene that co-segregates with a seed mutant phenotype. The mutant phenotype linked to this knockout generates novel hypotheses about the role for the plastid-localized oxidative pentose phosphate pathway during grain-fill. CONCLUSION: We show that FSTs from the UniformMu population identify stable, germinal insertion sites in maize. Moreover, we show that these sequence-indexed mutations can be readily used for reverse genetic analysis. We conclude from these data that the current collection of 1,882 non-redundant insertion sites from UniformMu provide a genome-wide resource for reverse genetics.
Asunto(s)
Bases de Datos Genéticas , Silenciador del Gen , Mutagénesis Insercional/métodos , Lugares Marcados de Secuencia , Zea mays/genética , Secuencia de Bases , Análisis por Conglomerados , Biología Computacional , Elementos Transponibles de ADN/genética , Datos de Secuencia Molecular , Fosfogluconato Deshidrogenasa/genética , Semillas/genética , Análisis de Secuencia de ADNRESUMEN
Flow cytometry was used to assess the variability of endoreduplication in endosperms of maize inbred lines. Little variation was found between midwestern dent types, and high levels of endoreduplication were observed in popcorns. Endoreduplication is different between inbred lines by 13-18 days after pollination, and flow cytometric analysis of ploidy level was feasible until 20 DAP. To study the genetic regulation of endoreduplication, four inbreds were crossed to B73 and developing endosperms from both parental, reciprocal F(1), and backcross generations were subjected to flow cytometric analysis. Three measurements of endoreduplication were calculated from these data and analyzed as quantitative genetic traits. Multiple models of trait inheritance were considered including triploid, diploid, sporophytic maternal, and maternal and paternal zygotic nuclear inheritance. Maternal zygotic effects, often considered a form of parental imprinting, and maternal sporophytic effects were detected. To test the feasibility of introgressing a high endoreduplication phenotype into a midwestern dent inbred line, a backcross population was generated from B73 x Sg18. Parental and progeny endoreduplication levels were compared and heritabilities assessed. The heritabilities calculated from these data generally agree with the values calculated in the larger crossing experiments.