Maternal Sporophyte Research Articles

Fertility loss in senescing Arabidopsis ovules is controlled by the maternal sporophyte via a NAC transcription factor triad

Flowers have a species-specific fertile period during which pollination and fertilization have to occur to initiate seed and fruit development. Unpollinated flowers remain receptive for mere hours in some species, and up to several weeks in others before flower senescence terminates fertility. As such, floral longevity is a key trait subject to both natural selection and plant breeding. Within the flower, the life span of the ovule containing the female gametophyte is decisive for fertilization and the initiation of seed development. Here, we show that unfertilized ovules in Arabidopsis thaliana undergo a senescence program that generates morphological and molecular hallmarks of canonical programmed cell death processes in the sporophytically derived ovule integuments. Transcriptome profiling of isolated aging ovules revealed substantial transcriptomic reprogramming during ovule senescence, and identified up-regulated transcription factors as candidate regulators of these processes. Combined mutation of three most-up-regulated NAC (NAM, ATAF1/2, and CUC2) transcription factors, NAP/ANAC029, SHYG/ANAC047, and ORE1/ANAC092, caused a substantial delay in ovule senescence and an extension of fertility in Arabidopsis ovules. These results suggest that timing of ovule senescence and duration of gametophyte receptivity are subject to genetic regulation controlled by the maternal sporophyte.

Embryo CHH hypermethylation is mediated by RdDM and is autonomously directed in Brassica rapa

BackgroundRNA-directed DNA methylation (RdDM) initiates cytosine methylation in all contexts and maintains asymmetric CHH methylation. Mature plant embryos show one of the highest levels of CHH methylation, and it has been suggested that RdDM is responsible for this hypermethylation. Because loss of RdDM in Brassica rapa causes seed abortion, embryo methylation might play a role in seed development. RdDM is required in the maternal sporophyte, suggesting that small RNAs from the maternal sporophyte might translocate to the developing embryo, triggering DNA methylation that prevents seed abortion. This raises the question of whether embryo hypermethylation is autonomously regulated by the embryo itself or influenced by the maternal sporophyte.ResultsHere, we demonstrate that B. rapa embryos are hypermethylated in both euchromatin and heterochromatin and that this process requires RdDM. Contrary to the current models, B. rapa embryo hypermethylation is not correlated with demethylation of the endosperm. We also show that maternal somatic RdDM is not sufficient for global embryo hypermethylation, and we find no compelling evidence for maternal somatic influence over embryo methylation at any locus. Decoupling of maternal and zygotic RdDM leads to successful seed development despite the loss of embryo CHH hypermethylation.ConclusionsWe conclude that embryo CHH hypermethylation is conserved, autonomously controlled, and not required for embryo development. Furthermore, maternal somatic RdDM, while required for seed development, does not directly influence embryo methylation patterns.

Abundant expression of maternal siRNAs is a conserved feature of seed development

Small RNAs are abundant in plant reproductive tissues, especially 24-nucleotide (nt) small interfering RNAs (siRNAs). Most 24-nt siRNAs are dependent on RNA Pol IV and RNA-DEPENDENT RNA POLYMERASE 2 (RDR2) and establish DNA methylation at thousands of genomic loci in a process called RNA-directed DNA methylation (RdDM). In Brassica rapa, RdDM is required in the maternal sporophyte for successful seed development. Here, we demonstrate that a small number of siRNA loci account for over 90% of siRNA expression during B. rapa seed development. These loci exhibit unique characteristics with regard to their copy number and association with genomic features, but they resemble canonical 24-nt siRNA loci in their dependence on RNA Pol IV/RDR2 and role in RdDM. These loci are expressed in ovules before fertilization and in the seed coat, embryo, and endosperm following fertilization. We observed a similar pattern of 24-nt siRNA expression in diverse angiosperms despite rapid sequence evolution at siren loci. In the endosperm, siren siRNAs show a marked maternal bias, and siren expression in maternal sporophytic tissues is required for siren siRNA accumulation. Together, these results demonstrate that seed development occurs under the influence of abundant maternal siRNAs that might be transported to, and function in, filial tissues.

Maternal regulation of seed growth and patterning in flowering plants.

The plant haploid generation is specified late in higher plant development, and post-meiotic haploid plant cells divide mitotically to produce a haploid gametophyte, in which a subset of cells differentiates into the gametes. The immediate mother of the angiosperm seed is the female gametophyte, also called the embryo sac. In most flowering plants the embryo sac is comprised of two kinds of gametes (egg and central cell) and two kinds of subsidiary cells (antipodals and synergids) all of which descend from a single haploid spore produced by meiosis. The embryo sac develops within a specialized organ of the flower called the ovule, which supports and controls many steps in the development of both the embryo sac and the seed. Double fertilization of the central cell and egg cell by the two sperm cells of a pollen grain produce the endosperm and embryo of the seed, respectively. The endosperm and embryo develop under the influence of their precursor gametes and the surrounding tissues of the ovule and the gametophyte. The final size and pattern of the angiosperm seed then is the result of complex interactions across multiple tissues of three different generations (maternal sporophyte, maternal gametophyte, and the fertilization products) and three different ploidies (haploid gametophyte, diploid parental sporophyte and embryo, and triploid endosperm).

Paternally Expressed Imprinted Genes under Positive Darwinian Selection in Arabidopsis thaliana.

Genomic imprinting is an epigenetic phenomenon where autosomal genes display uniparental expression depending on whether they are maternally or paternally inherited. Genomic imprinting can arise from parental conflicts over resource allocation to the offspring, which could drive imprinted loci to evolve by positive selection. We investigate whether positive selection is associated with genomic imprinting in the inbreeding species Arabidopsis thaliana. Our analysis of 140 genes regulated by genomic imprinting in the A. thaliana seed endosperm demonstrates they are evolving more rapidly than expected. To investigate whether positive selection drives this evolutionary acceleration, we identified orthologs of each imprinted gene across 34 plant species and elucidated their evolutionary trajectories. Increased positive selection was sought by comparing its incidence among imprinted genes with nonimprinted controls. Strikingly, we find a statistically significant enrichment of imprinted paternally expressed genes (iPEGs) evolving under positive selection, 50.6% of the total, but no such enrichment for positive selection among imprinted maternally expressed genes (iMEGs). This suggests that maternally- and paternally expressed imprinted genes are subject to different selective pressures. Almost all positively selected amino acids were fixed across 80 sequenced A. thaliana accessions, suggestive of selective sweeps in the A. thaliana lineage. The imprinted genes under positive selection are involved in processes important for seed development including auxin biosynthesis and epigenetic regulation. Our findings support a genomic imprinting model for plants where positive selection can affect paternally expressed genes due to continued conflict with maternal sporophyte tissues, even when parental conflict is reduced in predominantly inbreeding species.

Dividing a Maternal Pie among Half-Sibs: Genetic Conflicts and the Control of Resource Allocation to Seeds in Maize.

Resource allocation to offspring is the battleground for various intrafamilial conflicts. Understanding these conflicts requires knowledge of how the different actors (mother, siblings with different paternal genotypes) influence resource allocation. In angiosperms, allocation of resources to seeds happens postfertilization, and the paternally inherited genome in offspring can therefore influence resource allocation. However, the precise mode of resource allocation-and, in particular, the occurrence of sibling rivalry-has rarely been investigated in plants. In this article, we develop a new method for analyzing the resource-allocation traits of the different actors (maternal sporophyte and half-sibs) using data obtained from a large-scale diallel cross experiment in maize involving mixed hand pollination and color markers to assess seed weight of known paternity. We found strong evidence for the occurrence of sibling rivalry: resources invested in an ear were allocated competitively, and offspring with different paternal genotypes aggressively competed for these resources, entailing a measurable direct cost to the mother. We also show how resource allocation can be described for each genotype by two maternal traits (source effect, average sink responsiveness) and two offspring traits (ability to attract maternal resources, competitive ability toward siblings). We will discuss how these findings help to understand how genetic conflicts shape resource-allocation traits in angiosperms.

Evidence for parent-of-origin effects and interparental conflict in seeds of an ancient flowering plant lineage.

Theoretical and empirical studies have long connected the evolutionary innovation of endosperm, a genetically biparental product of a double fertilization process unique to flowering plants (angiosperms), to conflicting parental interests over offspring provisioning. Yet, none of these studies examined interparental conflict in representatives of any of the most ancient angiosperm lineages. We performed reciprocal interploidy crosses in the water lily Nymphaea thermarum, a member of one of the most ancient angiosperm lineages, Nymphaeales. We find that an excess of paternal genomes is associated with an increase in endosperm growth. By contrast, maternal ploidy negatively influences development or growth of all seed components, regardless of paternal genome dosage. Most relevant to the conflict over distribution of maternal resources, however, is that growth of the perisperm (seed storage tissue derived from the maternal sporophyte, found in all Nymphaeales) is unaffected by paternal genome dosage-ensuring maternal control of maternal resources. We conclude that the evolutionary transfer of embryo-nourishing function from a genetically biparental endosperm to a genetically maternal perisperm can be viewed as an effective maternal strategy to recapture control of resource distribution among progeny, and thus that interparental conflict has influenced the evolution of seed development in this ancient angiosperm lineage.

Maternal Effects on Seed and Seedling Phenotypes in Reciprocal F 1 Hybrids of the Common Bean (Phaseolus vulgaris L.).

Maternal control of seed size in the common bean provides an opportunity to study genotype-independent seed weight effects on early seedling growth and development. We set out to test the hypothesis that the early heterotrophic growth of bean seedlings is determined by both the relative amount of cotyledon storage reserves and the genotype of the seedling, provided the hybrid genotype could be fully expressed in the seedlings. The hypothesis was tested via comparison of seed weight and seedling growth phenotypes of small-seeded (wild, ~0.10 g) and large-seeded (landrace, ~0.55 g) parents and their reciprocal F1 hybrids. Akaike's Information Criteria were used to estimate growth parameters and identify the phenotypic model that best represented the data. The analysis presented here indicates that the hybrid embryo genotype is not fully expressed during both seed and seedling growth and development. The analysis presented here shows that seed growth and development are controlled by the sporophyte. The strong similarity in seed size and shape of the reciprocal hybrid seed with seeds of the maternal parents is evidence of this control. The analysis also indicates that since the maternal sporophyte controls seed size and therefore the amount of cotyledon reserves, the maternal sporophyte indirectly controls early seedling growth because the cotyledons are the primary nutrient source during heterotrophic growth. The most interesting and surprising results indicated that the maternal effects extended to the root architecture of the reciprocal hybrid seedlings. This phenomenon could not be explained by seed size, but by alterations in the control of the pattern of gene expression of the seedling, which apparently was set by a maternally controlled mechanism. Although seed weight increase was the main target of bean domestication, it also had positive repercussions on early-growth traits and stand establishment.

Cross-Talk Between Sporophyte and Gametophyte Generations Is Promoted by CHD3 Chromatin Remodelers inArabidopsis thaliana

Angiosperm reproduction requires the integrated development of multiple tissues with different genotypes. To achieve successful fertilization, the haploid female gametophytes and diploid ovary must coordinate their development, after which the male gametes must navigate through the maternal sporophytic tissues to reach the female gametes. After fertilization, seed development requires coordinated development of the maternal diploid integuments, the triploid endosperm, and the diploid zygote. Transcription and signaling factors contribute to communication between these tissues, and roles for epigenetic regulation have been described for some of these processes. Here we identify a broad role for CHD3 chromatin remodelers in Arabidopsis thaliana reproductive development. Plants lacking the CHD3 remodeler, PICKLE, exhibit various reproductive defects including abnormal development of the integuments, female gametophyte, and pollen tube, as well as delayed progression of ovule and embryo development. Genetic analyses demonstrate that these phenotypes result from loss of PICKLE in the maternal sporophyte. The paralogous gene PICKLE RELATED 2 is preferentially expressed in the endosperm and acts antagonistically with respect to PICKLE in the seed: loss of PICKLE RELATED 2 suppresses the large seed phenotype of pickle seeds. Surprisingly, the alteration of seed size in pickle plants is sufficient to determine the expression of embryonic traits in the seedling primary root. These findings establish an important role for CHD3 remodelers in plant reproduction and highlight how the epigenetic status of one tissue can impact the development of genetically distinct tissues.

The role of cytokinin in ovule development in Arabidopsis

The life cycle of higher plants alternates between the haploid gametophyte and diploid sporophyte. The female gametophyte (FG), surrounded by the sporophyte, develops within the ovule and orients along the chalazal/micropylar axis. This polarity is important in cell specification and development for both the ovule and FG. Previously, cytokinin was shown to act in the sporophytic tissue to regulate FG development.1,2 In the highlighted study,3 we further showed that enriched cytokinin signaling in chalaza, the central domain of the ovule, is required for the specification of the functional megaspore, which usually occurs in the chalazal-most megaspore after meiosis. The restricted cytokinin signaling in the chalaza is achieved by localized cytokinin biosynthesis and perception. Here, we discuss the implications of this and other studies for the understanding of the role of two-component signaling in FG development and the genetic and cellular interactions between gametophytic and sporophytic cells. Further, we show that cytokinin-deficient mutants display distorted cell morphology in the inner integument and elevated mitotic activity in the maternal sporophyte. These results suggest that cytokinin negatively regulates cell proliferation in the sporophytic tissues surrounding the developing FG, consistent with previous results indicating that cytokinin deficiency causes an increase in the number of cells in the embryos and consequently an enlarged seed size.

Cross talk between the sporophyte and the megagametophyte during ovule development

In seed plant ovules, the diploid maternal sporophytic generation embeds and sustains the haploid generation (the female gametophyte); thus, two independent generations coexist in a single organ. Many independent studies on Arabidopsis ovule mutants suggest that embryo sac development requires highly synchronized morphogenesis of the maternal sporophyte surrounding the gametophyte, since megagametogenesis is severely perturbed in most of the known sporophytic ovule development mutants. Which are the messenger molecules involved in the haploid-diploid dialogue? And furthermore, is this one way communication or is a feedback cross talk? In this review, we discuss genetic and molecular evidences supporting the presence of a cross talk between the two generations, starting from the first studies regarding ovule development and ending to the recently sporophytic identified genes whose expression is strictly controlled by the haploid gametophytic generation. We will mainly focus on Arabidopsis studies since it is the species more widely studied for this aspect. Furthermore, possible candidate molecules involved in the diploid-haploid generations dialogue will be presented and discussed.

The maternally expressed WRKY transcription factor TTG2 controls lethality in interploidy crosses of Arabidopsis.

The molecular mechanisms underlying lethality of F1 hybrids between diverged parents are one target of speciation research. Crosses between diploid and tetraploid individuals of the same genotype can result in F1 lethality, and this dosage-sensitive incompatibility plays a role in polyploid speciation. We have identified variation in F1 lethality in interploidy crosses of Arabidopsis thaliana and determined the genetic architecture of the maternally expressed variation via QTL mapping. A single large-effect QTL, DR. STRANGELOVE 1 (DSL1), was identified as well as two QTL with epistatic relationships to DSL1. DSL1 affects the rate of postzygotic lethality via expression in the maternal sporophyte. Fine mapping placed DSL1 in an interval encoding the maternal effect transcription factor TTG2. Maternal parents carrying loss-of-function mutations in TTG2 suppressed the F1 lethality caused by paternal excess interploidy crosses. The frequency of cellularization in the endosperm was similarly affected by both natural variation and ttg2 loss-of-function mutants. The simple genetic basis of the natural variation and effects of single-gene mutations suggests that F1 lethality in polyploids could evolve rapidly. Furthermore, the role of the sporophytically active TTG2 gene in interploidy crosses indicates that the developmental programming of the mother regulates the viability of interploidy hybrid offspring.

Genetic subtraction profiling identifies genes essential for Arabidopsisreproduction and reveals interaction between the female gametophyte and the maternal sporophyte

Genetic subtraction and expression profiling of wild-type Arabidopsis and a sporophytic mutant lacking an embryo sac identified 1,260 genes expressed in the embryo sac; a total of 527 genes were identified for their expression in ovules of mutants lacking an embryo sac.

Control of seed mass and seed yield by the floral homeotic gene APETALA2

APETALA2 (AP2) is best known for its role in the regulation of flower meristem and flower organ identity and development in Arabidopsis. We show here that AP2 also plays an important role in determining seed size, seed weight, and the accumulation of seed oil and protein. We demonstrate genetically that AP2 acts through the maternal sporophyte and endosperm genomes to control seed weight and seed yield. Thus, AP2 functions outside the boundaries of flower meristem and flower organ development to affect agronomically relevant traits in Arabidopsis.

Seed provisioning within holly fruits: test of the hierarchical model

Seed provisioning within holly Ilex aquifolium fruits was examined after a controlled pollination experiment to obtain full-sibs. The distribution of dry mass in stony endocarp and endosperm was determined according to the rank in the size hierarchy of the sibs within fruit. Resources were more equally distributed among sibs in the endocarps than in the endosperms, which might suggest sibling competition among endosperms and no evidences of competition in the tissue controlled by the maternal sporophyte. When the sibs are ranked by size, seed provisioning fit reasonably with the hierarchical model. Differences among consecutive sibs increased down the hierarchy, the second sib in the size hierarchy was less variable in resource acquisition than the others, and the disparity in the distribution of resources among sibs decreased with fruit size increases. These results seems to be in accordance with both kin selection model and hierarchical model, which indicates at the same time control of resource provisioning by the maternal plant and differences in vigour among sibs.

Maternal Effects of theshort integumentMutation on Embryo Development inArabidopsis

Maternal gene products deposited in an animal egg determine the polarity of embryonic axes and regulate embryonic cell–cell communication important for morphogenesis. Here we report the first maternal-effect embryo-defective mutation discovered in a plant. Recessive mutations in theSHORT INTEGUMENT(SIN1) gene inArabidopsiswere previously shown to influence ovule development and flowering time. Here we show that asin1mutation has a pronounced maternal effect on zygotic embryo development. A homozygoussin1mutant embryo is normal when nursed by asin1/+ heterozygous maternal sporophyte. Strikingly, asin1or asin1/+ embryo that is nursed by asin1homozygous maternal sporophyte develops morphogenetic defects in the apical–basal and radial axes. The defects resemble those seen in some zygotic-effect embryonic pattern formation mutants. These results imply that in maternal cells theSIN1gene either codes for or controls the production of a diffusible morphogen necessary for proper zygotic embryogenesis.

Selective forces in the emergence of the seed habit

The evolution of the seed is one of the major events in the history of land plants. In this paper, we consider the suite of characters that define the seed habit, and discuss the probable selective pressures that produced each character. Our major conclusion is that most characters are a direct consequence of the origin of heterospory and of natural selection for propagules with larger food reserves. Seeds are traditionally defined by the possession of integuments. However, some heterosporous pteridophytes possess integument-like structures. Therefore, integuments cannot explain the evolutionary success of seed plants. Rather, we believe that the decisive character in this success is related to pollination. Seed plants differ from other heterosporous lineages in the capture of microspores before dispersal of the 'megaspore'. Modern gymnosperms all possess mechanisms whereby the maternal sporophyte withholds resources from potential propagules that have not been pollinated and/or fertilized. This represents an increase in efficiency over Pteridophytic reproduction. Wind-pollination means the propagule is vulnerable to pathogens that mimic pollen, and pathogen pressures may have contributed to some seed characters.

Genetic analysis of conflicts arising during development of seeds in the Angiospermophyta

The genetic differences that exist among seeds developing on the same maternal sporophyte in the Angiospermophyta can lead to conflicts as the seeds absorb maternal resources. To analyse the outcome of these conflicts, a population-genetic model is developed to give first, second and third order conditions for initial increase of a mutant that affects interactions among relatives in a diploid, diallelic, random-mating population. Conflicts within families of developing seeds are then considered as a particular case of the model, by supposing that there exists a mutant which, when expressed in the endosperm, causes the seed to receive a greater share of maternal resources. Such seeds then have ‘over-consumer’ phenotypes, in contrast to their ‘under-consumer’ sibs. Conditions for initial increase of the mutant are obtained for nine major types of embryo sac and three kinds of gene expression. It is shown that all dominant mutants start to increase. However, the set of recessive mutants that can start to increase is restricted to those that give the over-consumer a large fitness advantage and, or, a low rate of loss of fitness to under-consumers. Behaviour of dosage-threshold mutants is more complicated, depending more on the number of megaspores that contribute to the embryo sac and the contribution of embryo sac nuclei to the endosperm. By using these results, an attempt is made to interpret some features of the genetic system of endosperm. Explanations for double fertilization lack the generality one would wish to account for such a ubiquitous feature in the Angiospermophyta. The double dosage of maternal genes arising from Polygonum -type embryo sac development has little effect on conflicts within families. However, the occurrence of tetrasporic types of endosperm can be seen as the outcome of selection on maternal sporophytes, which eliminates conflicts among their progeny.

Kin selection and conflict in seed maturation

There are four genetically distinct components in the developing seeds of flowering plants: maternal sporophyte, gametophyte, endosperm, and embryo. Each component can potentially influence the quantity or quality of nutrients provided to the embryo of its seed, thereby reducing the amount available to embryos in other seeds of that plant. The theory of kin selection predicts that each component will be selected to favor its own embryo over the other embryos to the extent that it is more closely related to its own. Under this criterion, an embryo should be selected to try to acquire more nutrients than the endosperm should be selected to provide, the endosperm should try to supply more than the gametophyte should, and the gametophyte more than the parent sporophyte. Evidence for this conflict of interests is found in the higher frequency of endopolyploidy, nutrient-absorbing haustoria, and food storage tissues in the embryo and endosperm than in the gametophyte of maternal tissues. This theory also suggests how the gametophyte, which is the nurse tissue of gymnosperm seeds, was displaced from this role in the flowering plants by an endosperm initiated by a secondary fertilization. “Neoteny” in the pro-angiosperms created conditions in which (1) an endosperm initiated by double fertilization would be more closely related to the embryo than is the gametophyte and (2) the endosperm would be formed early enough to be of significant aid to the embryo. If this theory is correct it (1) requires a different approach to the study of seed morphology and physiology, (2) increases the plausibility of arguments that flowering plants are a polyphyletic group, (3) provides evidence that parents cannot always control the outcome of conflict with their offspring, and (4) forges a conceptual link in our understanding of the evolution of social interactions in plants and animals.

METHODS OF BREEDING FOR OIL QUALITY IN RAPE

Reciprocal crosses were made between plants of Brassica napus with seed oil containing 40 and 0 per cent erucic acid. Gas liquid chromatographic analysis snowed an erucic acid content of 22 to 24 per cent in the crossed seeds. These results indicate that dominant gene action is absent and that fatty acid composition of the oil is controlled by the developing embryo. Thus the seed on an F1 plant constitutes an F2 population.Methods were developed to determine the fatty acid composition of oil from single seeds and from one cotyledon of an embryo. The analysis of a single cotyledon allows the genotype to be determined one generation earlier than if bulk samples are used, and the remainder of the seed may be grown into a normal plant. It is suggested that this half-seed technique may be used to determine the inheritance of other fatty acids in Brassica, and may also be applied to breeding other oil crops where the embryo, rather than the maternal sporophyte, controls the synthesis of fatty acids.