Endosperm sterol phenotype and germination in wheat

Free and conjugated sterols of endosperm, coats, scutellum, coleoptile and roots have been analysed at different germination stages in two wheat cultivars with different endosperm sterol phenotypes. It seems that sterol metabolism of the developing tissues, namely coleoptile and roots, is not affected by the sterol conjugation profile of the endosperm. Enough sterol is present in the mature embryo to supply the germinating axis during the observation period (144 hr at 16°). The data suggest that sterol is transferred from scutellum to coleoptile and roots during germinatio

The mechanics of Arabidopsis seed germination

Germination is defined as the protrusion of the embryonic radicle through the seed coat layers (endosperm and testa). As the radicle elongates, the testa ruptures, followed by rupture of the endosperm. Arabidopsis seeds exhibit a two-step germination process with sequential rupture of the testa and endosperm. We are interested in exploring the physical process of germination. Whilst much effort has previously been placed on genetic networks, a mathematical approach for furthering the understanding of the physical/mechanical properties of germination has not yet been described. The Mathematics in Plant Sciences Study Group helped us to develop a better understanding of the problem. Several different mathematical models were generated for radicle growth and endosperm stretching. These models were developed on multiscale dimensions – looking at the organ, tissue and cellular levels. The outcomes of the study group have heightened our interest in the mechanical aspects of germination, and we are currently progressing with a grant proposal – a collaboration between the Schools of Biosciences and Engineering at the University of Nottingham, and a group from the Department of Biology at the University of Freiburg, Germany

A novel superior factor widely controlling the rice grain quality

Synthesis of storage starch and protein accumulation is the main action of endosperm organogenesis in term of the economic importance of rice. This event is strongly disturbed by abiotic stresses such as high temperature; thus, the upcoming global warming will cause a crisis with a great impact on food production^1,2^. The enzymes for the protein storage and starch synthesis pathway should work in concert to carry out the organogenesis of rice endosperm^3-5^, but the regulatory mechanism is largely unknown. Here we show that a novel regulatory factor, named OsCEO1, acts as the conductor of endosperm organogenesis during the rice grain filling stage. The physiological properties of _floury-endosperm-2_ (_flo2_) mutants showed many similarities to symptoms of grains developed under high-temperature conditions, suggesting important roles of the responsible gene in sensitivity to high-temperature stress. Our map-based cloning identified the responsible gene for the _flo2_ mutant, _OsCEO1_, which has no homology to any genes of known function. The _OsCEO1_ belongs to a novel conserved gene family and encodes a protein composed of 1,720 amino acid residues containing a TPR (tetratricopeptide repeat) motif, which is considered to mediate a protein-protein interaction. The yeast two-hybrid analysis raised an unknown protein showing homology to a late embryogenesis abundant protein and a putative basic helix-loop-helix protein as candidates for the direct interactor for _OsCEO1_, whereas no enzyme genes for the synthesis of storage substances were detected. The _flo2_ mutant exhibited reduced expression of several genes for putative regulatory proteins as well as many enzymes involved in storage starch and proteins. These results suggest that _OsCEO1_ is a superior conductor of the novel regulatory cascade of endosperm organogenesis and may have important roles in the response to high-temperature stress

A general statistical framework for dissecting parent-of-origin effects underlying endosperm traits in flowering plants

Genomic imprinting has been thought to play an important role in seed development in flowering plants. Seed in a flowering plant normally contains diploid embryo and triploid endosperm. Empirical studies have shown that some economically important endosperm traits are genetically controlled by imprinted genes. However, the exact number and location of the imprinted genes are largely unknown due to the lack of efficient statistical mapping methods. Here we propose a general statistical variance components framework by utilizing the natural information of sex-specific allelic sharing among sibpairs in line crosses, to map imprinted quantitative trait loci (iQTL) underlying endosperm traits. We propose a new variance components partition method considering the unique characteristic of the triploid endosperm genome, and develop a restricted maximum likelihood estimation method in an interval scan for estimating and testing genome-wide iQTL effects. Cytoplasmic maternal effect which is thought to have primary influences on yield and grain quality is also considered when testing for genomic imprinting. Extension to multiple iQTL analysis is proposed. Asymptotic distribution of the likelihood ratio test for testing the variance components under irregular conditions are studied. Both simulation study and real data analysis indicate good performance and powerfulness of the developed approach.Comment: Published in at http://dx.doi.org/10.1214/09-AOAS323 the Annals of Applied Statistics (http://www.imstat.org/aoas/) by the Institute of Mathematical Statistics (http://www.imstat.org

Development and germination of Sandersonia aurantiaca (Hook.) seeds : a thesis presented in partial fulfilment of the requirements for the degree of Master of Science in Plant Biology and Biotechnology at Massey University

Sandersonia aurantiaca (Hook.) has recently become an important horticultural crop through its economic value for export of its cut flowers and tubers. Little information however is available on seed structure, morphology, development and propagation. The main objectives of this study were to investigate the pattern of seed development, to find satisfactory methods of improving the seed germination and to assess possible mechanisms of seed dormancy of Sandersonia aurantiaca (Hook.). Seed development was investigated by fixing plant material in FAA solution, embedding in paraffin, and staining with safranin-fast green. A series of sections were examined and photographed under a microscope. Both embryo and endosperm development in Sandersonia show close similarity to development in Allium fistulosum (Alliaceae). Embryo development passes through early globular, late globular, elongated spheroidal and linear embryo development stages. Endosperm development conforms to the Nuclear type. Freely-growing walls between the endosperm nuclei may be associated with the embryo sac wall as projections. The structure of the mature seeds is very similar to that of Iris (Iridaceae) seeds. The small, linear embryo is embedded in the endosperm which constitutes most of the seed volume. Such small, linear embryos may be one reason for embryo dormancy in Sandersonia seed. A special structure (a conical or cylindrical protuberance) is observed in the inner part of the seed coat, which may combine with a lignified layer (and perhaps including the endosperm) to contribute to the coat-imposed domancy in this species. Eighty five treatments were firstly used to improve the germination percentage of Sandersonia seed. Only the treatment in which seeds scarified firstly with sandpaper for 1 min and then nicked near the radicle end showed increased germination from 0 to 10.6% by 30 days, at 20°C. Based on this result, 31 new treatment methods were designed in germination experiment 2. Water uptake patterns, allelopathic effect on lettuce seeds and embryo rescue of Sandersonia seed were also studied for assessing the possible mechanisms of dormancy. The findings of the present study suggest that the Sandersonia seeds have double dormancy. The dormancy mechanism is located in both the seed coat and the embryo and it consists of at least two steps that must be activated in sequence before germination can occur. The first step can be activated prematurely by scarifying and nicking the seeds, thus allowing the seed coat to become permeable to water, oxygen or to reduced mechanical restriction. The second step can be activated directly GA3 which stimulates embryo growth. This germination-promoting technique has great potential for Sandersonia for improvement of the germination percentage of seeds from 0 to about 70%, but development on a commercial scale needs further studies

Storage lipids and proteins of Euterpe edulis seeds

Comparative studies on fatty acid and protein composition of the endosperm and embryo of palmito (Euterpe edulis Martius) were conducted using gas-liquid chromatography and sodium dodecyl sulfate–polyacrylamide gel electrophoresis. On a dry weight basis, the embryo contained extremely lower amounts of lipids and proteins than did the endosperm, which was associated with the scarce lipid and protein bodies previously reported in axis and cotyledon. The fatty acid composition also exhibited differences between both tissues: (I) the fatty acid diversity was greater in embryo than in endosperm; (II) embryo and endosperm contained predominantly linoleic, palmitic, oleic and stearic acids even though the relative values were different for each tissue. As compared to other palm species, the higher fatty acid unsaturation in Euterpe edulis seed could be involved in the previously reported short longevity and recalcitrant behavior during storage. Proteins of both tissues were heterogeneous in molecular mass. Some proteins were tissue-specific, but other were common, among them a highly glycosylated protein which migrated at about 55 kDa. We hypothesize that the latter, also reported in all previously studied palm species, is one of the proteins characterizing the Arecaceae family.Fil: Panza, Víctor. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación de Recursos Naturales. Instituto de Recursos Biológicos; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Biodiversidad y Biología Experimental; ArgentinaFil: Pighín, Darío Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; Argentina. Instituto Nacional de Tecnología Agropecuaria; ArgentinaFil: Lainez, Veronica. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; ArgentinaFil: Pollero, Ricardo Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Maldonado, Sara Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación de Recursos Naturales. Instituto de Recursos Biológicos; Argentin

Regulation of seed germination in the close Arabidopsis relative Lepidium sativum : a global tissue-specific transcript analysis

The completion of germination in Lepidium sativum and other endospermic seeds (e.g. Arabidopsis [Arabidopsis thaliana]) is regulated by two opposing forces, the growth potential of the radicle (RAD) and the resistance to this growth from the micropylar endosperm cap (CAP) surrounding it. We show by puncture force measurement that the CAP progressively weakens during germination, and we have conducted a time-course transcript analysis of RAD and CAP tissues throughout this process. We have also used specific inhibitors to investigate the importance of transcription, translation, and posttranslation levels of regulation of endosperm weakening in isolated CAPs. Although the impact of inhibiting translation is greater, both transcription and translation are required for the completion of endosperm weakening in the whole seed population. The majority of genes expressed during this process occur in both tissues, but where they are uniquely expressed, or significantly differentially expressed between tissues, this relates to the functions of the RAD as growing tissue and the CAP as a regulator of germination through weakening. More detailed analysis showed that putative orthologs of cell wall-remodeling genes are expressed in a complex manner during CAP weakening, suggesting distinct roles in the RAD and CAP. Expression patterns are also consistent with the CAP being a receptor for environmental signals influencing germination. Inhibitors of the aspartic, serine, and cysteine proteases reduced the number of isolated CAPs in which weakening developed, and inhibition of the 26S proteasome resulted in its complete cessation. This indicates that targeted protein degradation is a major control point for endosperm weakening

DNA demethylation by ROS1a in rice vegetative cells promotes methylation in sperm.

Epigenetic reprogramming is required for proper regulation of gene expression in eukaryotic organisms. In Arabidopsis, active DNA demethylation is crucial for seed viability, pollen function, and successful reproduction. The DEMETER (DME) DNA glycosylase initiates localized DNA demethylation in vegetative and central cells, so-called companion cells that are adjacent to sperm and egg gametes, respectively. In rice, the central cell genome displays local DNA hypomethylation, suggesting that active DNA demethylation also occurs in rice; however, the enzyme responsible for this process is unknown. One candidate is the rice REPRESSOR OF SILENCING 1a (ROS1a) gene, which is related to DME and is essential for rice seed viability and pollen function. Here, we report genome-wide analyses of DNA methylation in wild-type and ros1a mutant sperm and vegetative cells. We find that the rice vegetative cell genome is locally hypomethylated compared with sperm by a process that requires ROS1a activity. We show that many ROS1a target sequences in the vegetative cell are hypomethylated in the rice central cell, suggesting that ROS1a also demethylates the central cell genome. Similar to Arabidopsis, we show that sperm non-CG methylation is indirectly promoted by DNA demethylation in the vegetative cell. These results reveal that DNA glycosylase-mediated DNA demethylation processes are conserved in Arabidopsis and rice, plant species that diverged 150 million years ago. Finally, although global non-CG methylation levels of sperm and egg differ, the maternal and paternal embryo genomes show similar non-CG methylation levels, suggesting that rice gamete genomes undergo dynamic DNA methylation reprogramming after cell fusion

Ectopic application of the repressive histone modification H3K9me2 establishes post-zygotic reproductive isolation in Arabidopsis thaliana

Hybrid seed lethality as a consequence of interspecies or interploidy hybridizations is a major mechanism of reproductive isolation in plants. This mechanism is manifested in the endosperm, a dosage-sensitive tissue supporting embryo growth. Deregulated expression of imprinted genes such as ADMETOS (ADM) underpin the interploidy hybridization barrier in Arabidopsis thaliana; however, the mechanisms of their action remained unknown. In this study, we show that ADM interacts with the AT hook domain protein AHL10 and the SET domain-containing SU (VAR) 3-9 homolog SUVH9 and ectopically recruits the heterochromatic mark H3K9me2 to AT-rich transposable elements (TEs), causing deregulated expression of neighboring genes. Several hybrid incompatibility genes identified in Drosophila encode for dosage-sensitive heterochromatin-interacting proteins, which has led to the suggestion that hybrid incompatibilities evolve as a consequence of interspecies divergence of selfish DNA elements and their regulation. Our data show that imbalance of dosage-sensitive chromatin regulators underpins the barrier to interploidy hybridization in Arabidopsis, suggesting that reproductive isolation as a consequence of epigenetic regulation of TEs is a conserved feature in animals and plants

The catalytic core of DEMETER guides active DNA demethylation in Arabidopsis.

The Arabidopsis DEMETER (DME) DNA glycosylase demethylates the maternal genome in the central cell prior to fertilization and is essential for seed viability. DME preferentially targets small transposons that flank coding genes, influencing their expression and initiating plant gene imprinting. DME also targets intergenic and heterochromatic regions, but how it is recruited to these differing chromatin landscapes is unknown. The C-terminal half of DME consists of 3 conserved regions required for catalysis in vitro. We show that this catalytic core guides active demethylation at endogenous targets, rescuing dme developmental and genomic hypermethylation phenotypes. However, without the N terminus, heterochromatin demethylation is significantly impeded, and abundant CG-methylated genic sequences are ectopically demethylated. Comparative analysis revealed that the conserved DME N-terminal domains are present only in flowering plants, whereas the domain architecture of DME-like proteins in nonvascular plants mainly resembles the catalytic core, suggesting that it might represent the ancestral form of the 5mC DNA glycosylase found in plant lineages. We propose a bipartite model for DME protein action and suggest that the DME N terminus was acquired late during land plant evolution to improve specificity and facilitate demethylation at heterochromatin targets