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Pollen development and fertilization in Arabidopsis is dependent on the MALE GAMETOGENESIS IMPAIRED ANTHERS gene encoding a type V P-type ATPase.

In flowering plants, development of the haploid male gametophytes (pollen grains) takes place in a specialized structure called the anther. Successful pollen development, and thus reproduction, requires high secretory activity in both anther tissues and pollen. In this paper, we describe a novel member of the eukaryotic type V subfamily (P(5)) of P-type ATPase cation pumps, the MALE GAMETOGENESIS IMPAIRED ANTHERS (MIA) gene. MIA protein is highly abundant in the endoplasmic reticulum and small vesicles of developing pollen grains and tapetum cells. T-DNA insertional mutants of MIA suffer from imbalances in cation homeostasis and exhibit a severe reduction in fertility. Mutant microspores fail to separate from tetrads and pollen grains are fragile with an abnormal morphology and altered cell wall structure. Disruption of MIA affects expression of genes essential for secretion as well as a high number of genes encoding cell wall proteins and membrane transporters. MIA functionally complements a mutant in the P(5) ATPase homolog SPF1 from Saccharomyces cerevisiae, suggesting a common function for P(5) ATPases in single and multicellular organisms. Our results suggest that MIA is required in the secretory pathway for proper secretion of vesicle cargo to the plasma membrane.

ATP-Binding Cassette Transporters↗

[Effects of six host plants on the development and fecundity of Cicadulina bipunctella].

The study on the development, survivorship and fecundity of Cicadulina bipunctella reared on corn, wheat, sorghum, rice, millet, barley and Leptochloa panacea showed that the insect could develop and reproduce on all test host plants except for Leptochloa panacea. The development duration of C. bipunctella at its immature stages reared on sorghum was the shortest (24.1 d), then on corn (24.2 d), and the longest on wheat (25.5 d). The survival rate (40.8%), longevity (12.2 d) and oviposition per female (12.3 eggs) of female adults reared on rice were lower than those reared on the other five host plants. Based on the life-table parameters, the most suitable host plants for the development and reproduction of C. bipunctella were millet and corn, followed by sorghum, barley, wheat and rice.

Animals↗

FLP-mediated recombination for use in hybrid plant production.

We have studied the feasibility in Arabidopsis of using a site-specific recombination system FLP/FRT, from the 2 microm plasmid of yeast, for making plant hybrids. Initially, Arabidopsis plants expressing the FLP site-specific recombinase were crossed with plants transformed with a vector containing kanamycin-resistance gene (npt) flanked by FRT sites, which also served to separate the CaMV35S promoter from a promoterless gusA. Hybrid progeny were tested for excision of the npt gene and the positioning of 35S promoter proximal to gusA. GUS activity was observed in the progeny of all crosses, but not in the progeny derived from the self-pollinated homozygous parents. We then induced male sterility in Arabidopsis plants using the antisense expression of a pollen- and tapetum-specific gene, bcp1, flanked by FRT sites. Upon cross-pollination of flowers on the same male-sterile plants with pollen from FLP-containing plants, viable seeds were produced and the progeny hybrid plants developed normally. Molecular analyses revealed that the antisense expression cassette of bcp1 had been excised in these plants. These results show for the first time that a site-specific recombinase can be used to restore fertility in male-sterile plants, providing an alternative method for the production of hybrid seeds and plants.

Arabidopsis↗

Plant homologues of components of MAPK (mitogen-activated protein kinase) signal pathways in yeast and animal cells.

As they respond to numerous extracellular and intracellular stimuli, plants develop various morphological features and the capacity for a large variety of physiological processes during their growth. If we are to understand the molecular basis of such developments, we must elucidate the way in which signals generated by such stimuli can be transduced into plant cells and transmitted by cellular components to induce the appropriate terminal events. In yeast and animal systems, signal pathways that are known collectively as MAPK (mitogen-activated protein kinase) cascades have been shown to play a central role in the transmission of various signals. The components of these pathways include the MAPK family, the activator kinases of the MAPK family (the MAPKK family) and the activator kinases of the MAPKK family (the MAPKKK family). The members of each respective family are structurally conserved and signals are transmitted by similar phosphotransfer reactions at corresponding steps that are mediated by a specific member of each family in turn. Both cDNAs and genes that encode putative homologues of these components have recently been isolated from plant sources. Some of them have been shown to be related not only structurally but also functionally to members of the MAPK cascades of other organisms. These findings suggest that plants have signal pathways that are analogues to the MAPK cascades in yeast and animal cells but it remains to be proven that plant homologues do in fact constitute kinase cascades.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗

Characterization of tobacco MADS-box genes involved in floral initiation.

MADS-box genes encode regulatory factors that are involved at various stages in plant development. These genes function not only during early floral meristem identity, but also when the fate of floral organ primordia is determined in a later step. Here, we screened a floral bud cDNA library to isolate a tobacco MADS-box gene, NtMADS4, using the rice MADS-box gene, OsMADS1, as a probe. We previously reported that OsMADS1 plays a critical role in flower development in rice. Ectopic expression of NtMADS4 caused phenotypes of extremely early flowering as well as dwarfism. Plant MADS proteins have a K domain that mediates the formation of dimers. This dimerization appears to be an essential step for a functional protein complex. NtMADS11 was isolated as an interacting partner of NtMADS4 by yeast two-hybrid screening. The latter was included in the AGAMOUS-like 2 (AGL2) family whereas the former was categorized in the SQUAMOSA (SQUA) family. While the transcript of NtMADS4 was detectable only in reproductive organs, that of NtMADS11 was seen in both reproductive and vegetative organs. Expression levels were high for both genes during early developmental stages. Ectopic expression of NtMADS11 and OsMADS14 was able to rescue the floral organ defects seen in the strong ap1-1 mutant. Roles of NtMADS4 and NtMADS11 in the floral initiation are discussed.

Amino Acid Sequence↗

The HAT4 gene of Arabidopsis encodes a developmental regulator.

The HAT4 gene from the plant Arabidopsis thaliana encodes a homeo domain protein that contains a leucine zipper motif. Homeo domain-leucine zipper (HD-Zip) proteins have not been found in animal systems, suggesting that HAT4 may define a new family of transcription factors that regulate higher plant development. To explore this possibility, functional studies of HAT4 were carried out in yeast and in transgenic plants. Point mutants of HAT4 isolated in yeast define functionally critical residues within the HD-Zip domain, many of which correspond to highly conserved positions in known homeo domains and leucine zippers. Transgenic plants bearing constructs that alter HAT4 expression exhibit a series of interesting developmental phenotypes, including changes in morphology and developmental rate. Thus, the HAT4 gene of Arabidopsis encodes an HD-Zip protein that functions as a novel developmental regulator.

Amino Acid Sequence↗

Morphology of Arabidopsis Grown under Chronic Centrifugation and on the Clinostat.

Morphological measurements were made on populations of Arabidopsis thaliana grown from seed for 21 days under essentially constant environmental conditions except for the influence of gravitational or centrifugal accelerations. Growth conditions were what had been proposed for experiments in an artificial satellite. Observations are reported for plants grown at normal 1-g upright or on horizontal clinostats and for plants grown on a centrifuge. Increased g-force, up to 15 times normal, was found to have significant but small effects on some morphological end points. The plants' sensitivity to the magnitude of the g-force was much less than to its vector direction.Data from centrifuge experiments (which determined the g-functions for particular characters) were extrapolated to zero-g to predict a set of morphological characteristics of a plant developing in the satellite environment. As an alternative means of predicting properties of a zero-g plant, characteristics of plants grown on horizontal clinostats were measured. The results of these two predictive methods were not in agreement.Clinostat grown plants were morphologically distinct from upright stationary controls. When plants were grown while rotating in the upright position on vertical clinostats they were similar to stationary plants also grown upright, but there were small differences some of which were statistically significant.

Journal Article↗

Analysis of the transcription factor WUSCHEL and its functional homologue in Antirrhinum reveals a potential mechanism for their roles in meristem maintenance.

One of the most significant features of plant development is the way in which it can be elaborated and modulated throughout the life of the plant, an ability that is conferred by meristems. The Arabidopsis thaliana WUSCHEL gene (WUS), which encodes a homeodomain transcription factor, is required to maintain the stem cells in the shoot apical meristem in an undifferentiated state. The mechanism by which WUS prevents the differentiation of stem cells is unknown. We have characterized a meristem maintenance mutant in Antirrhinum majus and shown that it arises from a defect in the WUS orthologue ROSULATA (ROA). Detailed characterization of a semidominant roa allele revealed an essential role for the conserved C-terminal domain. Expression of either ROA or WUS lacking this domain causes a failure of meristem maintenance. The conserved domain mediates an interaction between WUS and two members of a small family of corepressor-like proteins in Arabidopsis. Our results suggest that WUS functions by recruiting transcriptional corepressors to repress target genes that promote differentiation, thereby ensuring stem cell maintenance.

Alleles↗

Dynamic of the genetic structure of bacterial and fungal communities at different developmental stages of Medicago truncatula Gaertn. cv. Jemalong line J5.

The genetic structure of bacterial and fungal communities was characterized in the rhizosphere of Medicago truncatula Gaertn. cv. Jemalong line J5 at five developmental stages (three vegetative and two reproductive stages), and in three compartments (bulk soil, rhizosphere soil and root tissues). The genetic structure of microbial communities was determined by cultivation-independent methods using directly extracted DNA that was characterized by automated ribosomal intergenic spacer analysis (ARISA). Principal component analyses (PCA) indicate that, for all developmental stages, the genetic structure of microbial communities differed significantly by compartment, with a major shift in the community in root tissues corresponding to the most intimate compartment with the plant. Differences were also recorded during plant development, the most significant being observed during the transition between vegetative and reproductive stages. Throughout this period, plants were shown to establish the highest level of symbiotic association (mycorrhization, nodulation) with arbuscular mycorrhizal fungi and Rhizobia. During the reproductive stages, the dynamics of the genetic structure differed between bacterial and fungal communities. At the last reproductive stage, the genetic structure of bacterial communities became close to that recorded during the first vegetative stages, suggesting a resilience phenomenon, whereas the genetic structure of fungal communities remained different from the vegetative stages and also from the early reproductive stages, suggesting a persistence of the rhizosphere effect.

Bacteria↗

[Hypoploid groups and their cytogenetic analysis in the progenies between Brassica napus and Orychophragmus violaceus].

Several groups of hypoploids were characterized in pentaploid (p) progenies of B. napus and O. violaceus. Cytological characterization of the P3 population revealed allosubstitution of one pair of B. napus chromosomes with O. violaceus chromosomes. This population had normal fertility and had three kinds of somatic cells with 36-38 chromosomes. Its somatic cells and pollen mother cells (PMC) mainly had 38 chromosomes. From a P4 population, three monosomic plants with 37 chromosomes were identified with differing morphology and fertility. One plant which grew vigorously was composed of various types of somatic cell and PMC, in which the cells with 37 chromosomes were observed mainly. The unpaired chromosome was small and with no negative effects on plant development. In the offsprings of the aneuploids (P3) with 41-44 chromosomes, there are four types of plants with 29-32 chromosomes. Except the plant with 29 chromosomes, others which appeared abnormal meiosis behavior and poor pollen fertility are mixoploid. The origin of these hypoploids and their prospects of application have been discussed.

Aneuploidy↗

Development of a quantitative tool for measuring changes in the coefficient of conductivity of plasmodesmata induced by developmental, biotic, and abiotic signals.

The regulation of intercellular and interorgan communication is pivotal for cell fate decisions in plant development and probably plays a significant role in the systemic regulation of gene expression and in defense reactions against pathogens or other biotic and abiotic environmental factors. In plants, symplasmic cell-to-cell communication is provided by plasmodesmata (Pd), coaxial membranous tunnels that span cell walls interconnecting adjacent cytoplasms. Macromolecules, proteins, and RNA may be transported through Pd by passive diffusion or by a facilitated mechanism. A quantitative tool was developed to measure the coefficient of conductivity, C(Pd), for diffusion-driven transport via Pd and to assess changes in the coefficient induced by developmental, biotic and abiotic signals. (GFP)C(Pd), the coefficient of conductivity for cell-to-cell spread of green-fluorescent protein (GFP), a protein with a Stokes radius of 2.82 nm, was determined in epidermal cells of sink and source leaves of wild-type and transgenic Nicotiana benthamiana plants expressing the movement protein of tobacco mosaic virus (MP(TMV)) incubated both in dark and light and at 16 and 25 degrees C. Under all conditions, Pd in source leaves conducted macromolecules, with (GFP)C(Pd)sink>(GFP)C(Pd)source. Light down-regulated (GFP)C(Pd) (all conditions); down-regulation was stronger for sink cells. The effect of MP(TMV) on (GFP)C(Pd) between epidermal cells was dependent on temperature and leaf development; at 16 degrees C, MP(TMV) down-regulated (GFP)C(Pd) only in source leaves, while at 25 degrees C, MP(TMV) had no significant effect. This quantitative tool should be useful for investigating differences in Pd conductivity that are induced by mutations or silencing.

Biological Transport, Active↗

Emerging from the fog: hypotheses and paradigms in developmental biology--the Society for Developmental Biology 2005 Annual Meeting Report.

The Society for Developmental Biology 64th annual meeting took place by the beautiful San Francisco Bay from July 27th to August 1st, 2005. Organized under the leadership of Judith Kimble (SDB President, U. Wisconsin-Madison), the meeting attracted over one thousand developmental biologists from all over the world. They gathered to present data, exchange ideas and enjoy basking in the warm sun on the piers. Strong themes emerged from the diverse subjects discussed at the meeting, demonstrating exciting trends towards the unifying goal of understanding the progression from a single cell to an adult organism. Cell and Tissue Polarity was a recurring topic at the meeting. Questions like "is there polarity", "how is it achieved" and "how is it linked to stem cell maintenance" were discussed. Post-transcriptional regulation involving protein degradation and microRNA (miRNA) modulation of gene expression was featured in the context of transition between meiosis to mitosis and asymmetries in the embryo. It is apparent that Evolutionary Developmental Biology, once a major driving influence in the early days of the field, continues to enjoy a renaissance as researchers familiar with traditional model organisms are increasingly attracted to the field and as modern genetic and molecular approaches are applied to an increasingly varied assortment of organisms. The attention is beginning to pay off as laboratories are starting to generate significant results shedding light into how developmental programs are altered to generate morphological diversity. In the Satellite Symposium on Plant Development held on July 27th, 2005, the overriding theme was on the identity and maintenance of Stem Cells in Plants. Finally, researchers working on diverse organisms have shown a strong effort to address Developmental Coordination: on the subcellular, cellular and tissue levels. Advanced imaging techniques are combined with traditional genetic methods to scrutinize and compare dynamic processes in four dimensions. This tremendous increase in resolution has facilitated the identification of key signaling mechanisms that embryos utilize to form coordinated body plans. For an exceptional effort in keeping with Society tradition, the 2005 annual meeting also offered opportunities to address broader issues revolving around education and professional development as well as a special session on embryonic stem cell research. Throughout the 5-day meeting, participants found time to honor the contributions of colleagues, exchange career and grant planning strategies, contemplate the big picture and recognize the efforts of young investigators, postdoctoral fellows and students.

Animals↗

Evaluation of wheat (Triticum aestivum L.) phenolic acids during grain development and their contribution to Fusarium resistance.

The phenolic acid profiles of six cultivars of wheat with known tolerance to Fusarium head blight were evaluated during plant development from anthesis through maturity. Analysis by HPLC of grain at anthesis revealed that p-coumaric and ferulic acid were the two principal phenolic compounds present. The effect of these two phenolic acids on Fusarium species was evaluated in vitro. Both phenolic acids demonstrated significant reductions (p < 0.05) of Fusarium species growth at all concentrations tested. Ferulic acid is the primary phenolic acid in grain at all stages of development, and its concentration increased steadily during grain development prior to a 50% decrease during grain ripening. The accumulation of ferulic acid synthesis from anthesis until approximately 20 days after anthesis appears related with cultivar resistance to Fusarium. Concentrations of ferulic acid in the grain were similar at maturity, implying that the end-use quality would be similar for both resistant and susceptible cultivars.

Coumaric Acids↗

Mutations, epimutations, and the developmental programming of the maize Suppressor-mutator transposable element.

Information about the structure, function and regulation of the maize Suppressor-mutator (Spm) transposable element has emerged from the genetic and molecular characterization of both deletion mutations and an unconventional type of reversible genetic change (epimutation). The element is subject to an epigenetic mechanism that can either stably inactivate it or specify one of a variety of heritable programs of differential element expression in development. The essay explores the relationship between the Spm element's epigenetic developmental programming mechanism and the determinative events central to plant development and differentiation.

Base Sequence↗

cDNA microarray analysis of small plant tissue samples using a cDNA tag target amplification protocol.

Microarray technology is becoming an important comprehensive tool to study gene expression in plants. However, the use of this technology is limited by the large amount of sample tissue needed for microarray analysis. Generally, 50-200 microg of total RNA and 1-2 microg of mRNA is required for each hybridisation, which is equivalent to 50-100 mg of plant tissue. This requirement for large amounts of starting material severely constrains the use of microarrays for transcript profiling in specific tissues and cell types during plant development. Here we report on a robust and reliable target amplification method that enables transcript profiling from sub-mg amounts of plant tissue. Using 0.1 microg of total RNA we show that twofold expression differences are possible to distinguish with 99% confidence. We also demonstrate the application of this method in an analysis of secondary phloem development in hybrid aspen using defined tissue sections, corresponding to 2-4 cell layers with a fresh weight of approximately 0.5 mg.

DNA, Complementary↗

Gene profiling of the red light signalling pathways in roots.

Red light, acting through the phytochromes, controls numerous aspects of plant development. Many of the signal transduction elements downstream of the phytochromes have been identified in the aerial portions of the plant; however, very few elements in red-light signalling have been identified specifically for roots. Gene profiling studies using microarrays and quantitative Real-Time PCR were performed to characterize gene expression changes in roots of Arabidopsis seedlings exposed to 1 h of red light. Several factors acting downstream of phytochromes in red-light signalling in roots were identified. Some of the genes found to be differentially expressed in this study have already been characterized in the red-light-signalling pathway for whole plants. For example, PHYTOCHROME KINASE 1 (PKS1), LONG HYPOCOTYL 5 (HY5), EARLY FLOWERING 4 (ELF4), and GIGANTEA (GI) were all significantly up-regulated in roots of seedlings exposed to 1 h of red light. The up-regulation of SUPPRESSOR OF PHYTOCHROME A RESPONSES 1 (SPA1) and CONSTITUTIVE PHOTOMORPHOGENIC 1-like (COP1-like) genes suggests that the PHYA-mediated pathway was attenuated by red light. In addition, genes involved in lateral root and root hair formation, root plastid development, phenylpropanoid metabolism, and hormone signalling were also regulated by exposure to red light. Interestingly, members of the RPT2/NPH3 (ROOT PHOTOTROPIC 2/NON PHOTOTROPIC HYPOCOTYL 3) family, which have been shown to mediate blue-light-induced phototropism, were also differentially regulated in roots in red light. Therefore, these results suggest that red and blue light pathways interact in roots of seedlings and that many elements involved in red-light-signalling found in the aerial portions of the plant are differentially expressed in roots within 1 h of red light exposure.

ATP-Binding Cassette Transporters↗

Gene expression of stearoyl-ACP desaturase and delta12 fatty acid desaturase 2 is modulated during seed development of flax (Linum usitatissimum).

Flax's recent popularity in human and animal foods is mostly due to its desirable FA composition. Flax is an excellent source of omega-3 FA, which have been shown to have many health benefits. To date, little is known about the genetic and environmental factors that control the FA composition of flax seeds. To elucidate some of the important genetic components, reverse transcriptase (RT)-PCR and real-time PCR were used to determine the expression profiles of two key FA biosynthetic genes during seed development. Plants of flax cultivar AC McDuff were grown under field conditions, and RNA was extracted from ovaries and developing bolls collected from 2 d after anthesis (DAA) to maturity. Desaturation enzymes stearoyl-ACP desaturase (SAD) and delta12 FA desaturase 2 (FAD2) were both expressed in ovaries, and their expression was differentially modulated throughout seed development. SAD was most highly expressed in ovaries. Its expression quickly decreased until 4 DAA; this was followed by a slight peak at 8 DAA, only to return to relatively low levels of expression in maturing bolls, ranging from 2.1% to 4.5% relative to the level observed in ovaries. FAD2 expression displayed a different temporal pattern. While expression of FAD2 did decrease in the early stages of seed development, expression increased starting at 8 DAA, peaking at 16 DAA, when it was 158% relative to the level observed in ovaries. FAD2, which desaturates oleic acid (18:1cisdelta9) into linoleic acid (18:2cisdelta9,12), is therefore controlled at the transcription level. To relate enzyme expression with FA profile, GC was performed on the same subsamples used for RT-PCR and real-time PCR, and proportions of palmitic, stearic, oleic, linoleic, and linolenic acids were determined for the same developmental stages. Although FAD2 expression increased from 8 to 16 DAA, relative changes in linoleic acid (18:2cis delta9,12) were not observed. However, linolenic acid (ALA; alpha-18:3; 18:3cisdelta9,12,15) levels increased steadily, meaning that linoleic acid (18:2cisdelta9,12) is a transient substrate converted by FAD3 as quickly as it is produced by FAD2. Phenotypes are the result of genotypes, environment, and the interaction of the two. To evaluate the environmental impact on the production of FA in flax, FA profiles were assessed in a total of four environments (two locations, two years). Warm and dry environmental conditions resulted in lower levels of PUFA 18:2cisdelta9,12 and 18:3cisdelta9,12,15, and higher levels of 18:1 cisdelta9. FAD2 expression and/or activity may therefore be affected by the environment.

Base Sequence↗

Dynamics of the genetic regulatory network for Arabidopsis thaliana flower morphogenesis.

We present a network model and its dynamic analysis for the regulatory relationships among 11 genes that participate in Arabidopsis thaliana flower morphogenesis. The topology of the network and the relative strengths of interactions among these genes were based from published genetic and molecular data, mainly relying on mRNA expression patterns under wild type and mutant backgrounds. The network model is made of binary elements and we used a particular dynamic implementation for the network that we call semi-synchronic. Using this method the network reaches six attractors; four of them correspond to observed patterns of gene expression found in the floral organs of Arabidopsis (sepals, petals, stamens and carpels) as predicted by the ABC model of flower morphogenesis. The fifth state corresponds to cells that are not competent to flowering, and the sixth attractor predicted by the model is never found in wild-type plants, but it could be induced experimentally. We discuss the biological implications and the potential use of this network modeling approach to integrate functional data of regulatory genes of plant development.

Arabidopsis↗