Flowering In Cereals Research Articles

OsFTL4, an FT-like Gene, Regulates Flowering Time and Drought Tolerance in Rice (Oryza sativa L.)

The initiation of flowering in cereals is a critical process influenced by environmental and endogenous signals. Flowering Locus T-like (FT-like) genes encode the main signals for flowering. Of the 13 FT-like genes in the rice genome, Hd3a/OsFTL2 and RFT1/OsFTL3 have been extensively studied and revealed to be critical for flowering. In this study, a rice FT-like gene, OsFTL4, was functionally characterized. Specifically, osftl4 mutants were generated using a CRISPR/Cas9 system. Compared with the wild-type control (Guangluai 4), the osftl4-1 and osftl4-2 mutants flowered 9.6 and 5.8 days earlier under natural long-day and short-day conditions, respectively. Additionally, OsFTL4 was mainly expressed in the vascular tissue, with the resulting OsFTL4 protein localized in both the nucleus and cytoplasm. Furthermore, OsFTL4 was observed to compete with Hd3a for the interaction with multiple 14-3-3 proteins. An analysis of the effects of simulated drought stress suggested that silencing OsFTL4 enhances drought tolerance by decreasing stomatal conductance and water loss. These results indicate that OsFTL4 helps integrate the flowering process and the drought response in rice.

Heat stress during flowering in cereals - effects and adaptation strategies.

Heat stress during flowering has differential impact on male and female reproductive organ viability leading to yield losses in field crops. Unlike flooded rice, dryland cereals such as sorghum, pearl millet and wheat have optimised their flower opening during cooler early morning or late evening hours to lower heat stress damage during flowering. Although previous studies have concluded that pollen viability determines seed set under heat stress, recent findings have revealed pearl millet and sorghum pistils to be equally sensitive to heat stress. Integrating flower opening time during cooler hours with increased pollen and pistil viability will overcome heat stress-induced damage during flowering under current and future hotter climatic conditions.

Zebularine treatment is associated with deletion of FT-B1 leading to an increase in spikelet number in bread wheat.

The number of rachis nodes (spikelets) on a wheat spike is a component of grain yield that correlates with flowering time. The genetic basis regulating flowering in cereals is well understood, but there are reports that flowering time can be modified at a high frequency by selective breeding, suggesting that it may be regulated by both epigenetic and genetic mechanisms. We investigated the role of DNA methylation in regulating spikelet number and flowering time by treating a semi-spring wheat with the demethylating agent, Zebularine. Three lines with a heritable increase in spikelet number were identified. The molecular basis for increased spikelet number was not determined in 2 lines, but the phenotype showed non-Mendelian inheritance, suggesting that it could have an epigenetic basis. In the remaining line, the increased spikelet phenotype behaved as a Mendelian recessive trait and late flowering was associated with a deletion encompassing the floral promoter, FT-B1. Deletion of FT-B1 delayed the transition to reproductive growth, extended the duration of spike development, and increased spikelet number under different temperature regimes and photoperiod. Transiently disrupting DNA methylation can generate novel flowering behaviour in wheat, but these changes may not be sufficiently stable for use in breeding programs.

The Importance of Being on Time: Regulatory Networks Controlling Photoperiodic Flowering in Cereals.

Flowering is the result of the coordination between genetic information and environmental cues. Gene regulatory networks have evolved in plants in order to measure diurnal and seasonal variation of day length (or photoperiod), thus aligning the reproductive phase with the most favorable season of the year. The capacity of plants to discriminate distinct photoperiods classifies them into long and short day species, depending on the conditions that induce flowering. Plants of tropical origin and adapted to short day lengths include rice, maize, and sorghum, whereas wheat and barley were originally domesticated in the Fertile Crescent and are considered long day species. In these and other crops, day length measurement mechanisms have been artificially modified during domestication and breeding to adapt plants to novel areas, to the extent that a wide diversity of responses exists within any given species. Notwithstanding the ample natural and artificial variation of day length responses, some of the basic molecular elements governing photoperiodic flowering are widely conserved. However, as our understanding of the underlying mechanisms improves, it becomes evident that specific regulators exist in many lineages that are not shared by others, while apparently conserved components can be recruited to novel functions during evolution.

Visual examination of changes in soil structural quality due to land use

This study aims to assess how responsive visual examination methods are to the effect of land use on soil structural quality (SSQ), and whether they are sensitive enough to detect significant changes on SSQ over a given sampling interval. The visual soil assessment (VSA), the visual evaluation of soil structure (VESS), the visual assessment of aggregate stability (VAAS) and the visual type of aggregates index (Tyagg) were used to evaluate the SSQ of a sandy loam and a silt loam soil. The land uses comprised cropland (CP) and grassland (PP). The survey was conducted twice in an agricultural cycle (in August and November). Results showed that VESS and Tyagg were in agreement with soil physical parameters when evaluating structural quality of a sandy loam, in contrast to VSA and VAAS. In the silt loam, all methods were responsive to land use effects on soil quality and sensitive in detecting changes in SSQ between evaluation times. We further showed that soils under PP resulted in the best SSQ compared to CP after harvesting, whereas SSQ of CP was better during cereal flowering than after harvesting. Despite the majority of the visual examination methods used in this survey were responsive in evaluating the effect of land use on SSQ and capable of representing structural dynamics (related to soil management) in an agricultural cycle, the lack of agreement between visual examinations and their interrelationships with the soil physical properties evaluated, however, highlight the need to conduct further work for exploring: a) method limitations, b) key factors such as soil moisture content and minimum number of samples for visual examination according to soil texture and spatial variability, and c) a judicious selection of a minimum data set of SSQ indicators omitting redundant material.

ODDSOC2 Is a MADS Box Floral Repressor That Is Down-Regulated by Vernalization in Temperate Cereals

In temperate cereals, such as wheat (Triticum aestivum) and barley (Hordeum vulgare), the transition to reproductive development can be accelerated by prolonged exposure to cold (vernalization). We examined the role of the grass-specific MADS box gene ODDSOC2 (OS2) in the vernalization response in cereals. The barley OS2 gene (HvOS2) is expressed in leaves and shoot apices but is repressed by vernalization. Vernalization represses OS2 independently of VERNALIZATION1 (VRN1) in a VRN1 deletion mutant of einkorn wheat (Triticum monococcum), but VRN1 is required to maintain down-regulation of OS2 in vernalized plants. Furthermore, barleys that carry active alleles of the VRN1 gene (HvVRN1) have reduced expression of HvOS2, suggesting that HvVRN1 down-regulates HvOS2 during development. Overexpression of HvOS2 delayed flowering and reduced spike, stem, and leaf length in transgenic barley plants. Plants overexpressing HvOS2 showed reduced expression of barley homologs of the Arabidopsis (Arabidopsis thaliana) gene FLOWERING PROMOTING FACTOR1 (FPF1) and increased expression of RNase-S-like genes. FPF1 promotes floral development and enhances cell elongation, so down-regulation of FPF1-like genes might explain the phenotypes of HvOS2 overexpression lines. We present an extended model of the genetic pathways controlling vernalization-induced flowering in cereals, which describes the regulatory relationships between VRN1, OS2, and FPF1-like genes. Overall, these findings highlight differences and similarities between the vernalization responses of temperate cereals and the model plant Arabidopsis.

The central role of the VERNALIZATION1 gene in the vernalization response of cereals

Many varieties of wheat (Triticum spp.) and barley (Hordeum vulgare L.) require prolonged exposure to cold during winter in order to flower (vernalization). In these cereals, vernalization-induced flowering is controlled by the VERNALIZATION1 (VRN1) gene. VRN1 is a promoter of flowering that is activated by low temperatures. VRN1 transcript levels increase gradually during vernalization, with longer cold treatments inducing higher expression levels. Elevated VRN1 expression is maintained in the shoot apex and leaves after vernalization, and the level of VRN1 expression in these organs determines how rapidly vernalized plants flower. Some alleles of VRN1 are expressed without vernalization due to deletions or insertions within the promoter or first intron of the VRN1 gene. Varieties of wheat and barley with these alleles flower without vernalization and are grown where vernalization does not occur. The first intron of the VRN1 locus has histone modifications typically associated with the maintenance of an inactive chromatin state, suggesting this region is targeted by epigenetic mechanisms that contribute to repression of VRN1 before winter. Other mechanisms are likely to act elsewhere in the VRN1 gene to mediate low-temperature induction. This review examines how understanding the mechanisms that regulate VRN1 provides insights into the biology of vernalization-induced flowering in cereals and how this will contribute to future cereal breeding strategies.

Association Studies Identify Natural Variation at PHYC Linked to Flowering Time and Morphological Variation in Pearl Millet

The identification of genes selected during and after plant domestication is an important research topic to enhance knowledge on adaptative evolution. Adaptation to different climates was a key factor in the spread of domesticated crops. We conducted a study to identify genes responsible for these adaptations in pearl millet and developed an association framework to identify genetic variations associated with the phenotype in this species. A set of 90 inbred lines genotyped using microsatellite loci and AFLP markers was used. The population structure was assessed using two different Bayesian approaches that allow inbreeding or not. Association studies were performed using a linear mixed model considering both the population structure and familial relationships between inbred lines. We assessed the ability of the method to limit the number of false positive associations on the basis of the two different Bayesian methods, the number of populations considered and different morphological traits while also assessing the power of the methodology to detect given additive effects. Finally, we applied this methodology to a set of eight pearl millet genes homologous to cereal flowering pathway genes. We found significant associations between several polymorphisms of the pearl millet PHYC gene and flowering time, spike length, and stem diameter in the inbred line panel. To validate this association, we performed a second association analysis in a different set of pearl millet individuals from Niger. We confirmed a significant association between genetic variation in this gene and these characters.

Vernalization-induced flowering in cereals is associated with changes in histone methylation at the VERNALIZATION1 gene

Prolonged exposure to low temperatures (vernalization) accelerates the transition to reproductive growth in many plant species, including the model plant Arabidopsis thaliana and the economically important cereal crops, wheat and barley. Vernalization-induced flowering is an epigenetic phenomenon. In Arabidopsis, stable down-regulation of FLOWERING LOCUS C (FLC) by vernalization is associated with changes in histone modifications at FLC chromatin. In cereals, the vernalization response is mediated by stable induction of the floral promoter VERNALIZATION1 (VRN1), which initiates reproductive development at the shoot apex. We show that in barley (Hordeum vulgare), repression of HvVRN1 before vernalization is associated with high levels of histone 3 lysine 27 trimethylation (H3K27me3) at HvVRN1 chromatin. Vernalization caused increased levels of histone 3 lysine 4 trimethylation (H3K4me3) and a loss of H3K27me3 at HvVRN1, suggesting that vernalization promotes an active chromatin state at VRN1. Levels of these histone modifications at 2 other flowering-time genes, VERNALIZATION2 and FLOWERING LOCUS T, were not altered by vernalization. Our study suggests that maintenance of an active chromatin state at VRN1 is likely to be the basis for epigenetic memory of vernalization in cereals. Thus, regulation of chromatin state is a feature of epigenetic memory of vernalization in Arabidopsis and the cereals; however, whereas vernalization-induced flowering in Arabidopsis is mediated by epigenetic regulation of the floral repressor FLC, this phenomenon in cereals is mediated by epigenetic regulation of the floral activator, VRN1.

The molecular biology of seasonal flowering-responses in Arabidopsis and the cereals

In arabidopsis (Arabidopsis thaliana), FLOWERING LOCUS T (FT) and FLOWERING LOCUS C (FLC) play key roles in regulating seasonal flowering-responses to synchronize flowering with optimal conditions. FT is a promoter of flowering activated by long days and by warm conditions. FLC represses FT to delay flowering until plants experience winter. The identification of genes controlling flowering in cereals allows comparison of the molecular pathways controlling seasonal flowering-responses in cereals with those of arabidopsis. The role of FT has been conserved between arabidopsis and cereals; FT-like genes trigger flowering in response to short days in rice or long days in temperate cereals, such as wheat (Triticum aestivum) and barley (Hordeum vulgare). Many varieties of wheat and barley require vernalization to flower but FLC-like genes have not been identified in cereals. Instead, VERNALIZATION2 (VRN2) inhibits long-day induction of FT-like1 (FT1) prior to winter. VERNALIZATION1 (VRN1) is activated by low-temperatures during winter to repress VRN2 and to allow the long-day response to occur in spring. In rice (Oryza sativa) a VRN2-like gene Ghd7, which influences grain number, plant height and heading date, represses the FT-like gene Heading date 3a (Hd3a) in long days, suggesting a broader role for VRN2-like genes in regulating day-length responses in cereals. Other genes, including Early heading date (Ehd1), Oryza sativa MADS51 (OsMADS51) and INDETERMINATE1 (OsID1) up-regulate Hd3a in short days. These genes might account for the different day-length response of rice compared with the temperate cereals. No genes homologous to VRN2, Ehd1, Ehd2 or OsMADS51 occur in arabidopsis. It seems that different genes regulate FT orthologues to elicit seasonal flowering-responses in arabidopsis and the cereals. This highlights the need for more detailed study into the molecular basis of seasonal flowering-responses in cereal crops or in closely related model plants such as Brachypodium distachyon.

Integration of seasonal flowering time responses in temperate cereals

Our paper describes how the genetic pathways which regulate vernalization and long-day flowering responses are integrated to promote spring flowering in cereals. This process is mediated by the VERNALIZATION1 (VRN1) and VRN2 genes. VRN2 encodes a CONSTANS-like protein that represses FLOWERING LOCUS T (FT1) to block the long-day flowering response until plants are vernalized. When plants are vernalized VRN1, a FRUITFUL-like MADS box transcription factor, is induced. This down-regulates VRN2, allowing long-day induction of FT1 to occur post-vernalization. A comparison of the pathways regulating seasonal induction of flowering in cereals with those of Arabidopsis shows the vernalization response pathway has evolved convergently to regulate the activity of a conserved daylength response pathway in these divergent plant lineages.Addendum to: Hemming M.N, Peacock W.J, Dennis E.S, Trevaskis B. Low temperature and daylength cues are integrated to regulate FLOWERING LOCUS T in barley. Plant Physiol 2008; 147:355-66.

Control of flowering time in temperate cereals: genes, domestication, and sustainable productivity

The control of flowering is central to reproductive success in plants, and has a major impact on grain yield in crop species. The global importance of temperate cereal crops such as wheat and barley has meant emphasis has long been placed on understanding the genetics of flowering in order to enhance yield. Leads gained from the dissection of the molecular genetics of model species have combined with comparative genetic approaches, recently resulting in the isolation of the first flowering time genes in wheat and barley. This paper reviews the genetics and genes involved in cereal flowering pathways and the current understanding of how two of the principal genes, Vrn and Ppd, have been involved in domestication and adaptation to local environments, and the implications for future breeding programmes are discussed.

Quantitative trait loci controlling vernalisation requirement, heading time and number of panicles in meadow fescue (Festuca pratensis Huds.)

The "BF14/16xHF2/7" mapping population of meadow fescue (Festuca pratensis Huds.) was characterised for number of panicles produced by non-vernalised plants in the field, vernalisation requirement (number of weeks at 6 degrees C and 8 h photoperiod), as well as days to heading, number of panicles and proportion of shoots heading after a 12 weeks vernalisation treatment. Quantitative trait loci (QTLs) were identified and compared to QTLs and genes related to the induction of flowering in cereals and grasses. A region on chromosome 1F affected days to heading and the proportion of shoots heading. Chromosome 4F appeared to have several genes with a strong effect on vernalisation requirement. The strongest effects were located in the proximal end of 4F and may correspond to the earliness per se (eps) QTL eps6L.2 in barley and a heading time QTL in perennial ryegrass. A part of the meadow fescue orthologue of VRN1 was sequenced and mapped to another region of 4F that also had a strong effect on vernalisation requirement. The proximal end of chromosome 5F had QTLs for days to heading and proportion of heading shoots. Syntenic regions in wheat and barley contain eps-loci. A QTL for number of panicles in the field and a QTL for proportion of heading shoots were present on chromosome 6. A region on 7F affected the variation in number of panicles among plants without a vernalisation requirement, and is syntenic to regions in perennial ryegrass, barley and rice containing orthologues of Arabidopsis thaliana CO.

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MADS box genes control vernalization-induced flowering in cereals.

By comparing expression levels of MADS box transcription factor genes between near-isogenic winter and spring lines of bread wheat, Triticum aestivum, we have identified WAP1 as the probable candidate for the Vrn-1 gene, the major locus controlling the vernalization flowering response in wheat. WAP1 is strongly expressed in spring wheats and moderately expressed in semispring wheats, but is not expressed in winter wheat plants that have not been exposed to vernalization treatment. Vernalization promotes flowering in winter wheats and strongly induces expression of WAP1. WAP1 is located on chromosome 5 in wheat and, by synteny with other cereal genomes, is likely to be collocated with Vrn-1. These results in hexaploid bread wheat cultivars extend the conclusion made by Yan et al. [Yan, L., Loukoianov, A., Tranquilli, G., Helguera, M., Fahima, T. & Dubcovsky, J. (2003) Proc. Natl. Acad. Sci. USA 100, 6263-6268] in the diploid wheat progenitor Triticum monococcum that WAP1 (TmAP1) corresponds to the Vrn-1 gene. The barley homologue of WAP1, BM5, shows a similar pattern of expression to WAP1 and TmAP1. BM5 is not expressed in winter barleys that have not been vernalized, but as with WAP1, expression of BM5 is strongly induced by vernalization treatment. In spring barleys, the level of BM5 expression is determined by interactions between the Vrn-H1 locus and a second locus for spring habit, Vrn-H2. There is now evidence that AP1-like genes determine the time of flowering in a range of cereal and grass species.

The Control of Flowering in Wheat and Barley: What Recent Advances in Molecular Genetics Can Reveal

The combined forces of developmental biologists, studying primordium initiation at the stem apex, and mathematical modellers, developing simulations of crop growth and development, have brought about considerable advances in the understanding of the control of flowering in wheat and barley. Nevertheless, there are still major gaps in this understanding including: what determines the basic rate of development (magnitude of the phyllochron or plastochron); how temperature and photoperiod interact to bring about the transition from vegetative to reproductive development; and how flowering occurs eventually in the absence of inductive conditions. Although geneticists have tended to measure cereal flowering in terms of ‘days from sowing or emergence to heading’, results of studies using aneuploids and molecular markers are compatible with the roles for photoperiod and low-temperature vernalization established in purely-physiological or developmental investigations. They have also revealed the existence of ‘earliness per se ’loci, whose detailed roles have yet to be established. Progress towards isolating and characterizing wheat and barley loci is hampered by the poor resolution of mapping (location to a precision of tens of thousands of base pairs). Neither of these broad approaches promises a rapid resolution of the factors controlling the induction of flowering. Two expanding areas of molecular genetics now provide potential for greater understanding of cereal flowering. First, the extensive homoeology among members of the Gramineae can be employed to establish the existence and location of genes or quantitative trait loci in rice which correspond to controlling loci in wheat or barley. Since the rice genome is 1/30th of the size of the wheat genome, the accuracy of mapping loci can be much higher, and there is greater potential for precise location of loci using techniques such as chromosome walking. With the ultimate cloning of individual genes, and the isolation of gene products, the relative roles of the 20 loci apparently involved in the induction of flowering of wheat could be explored. However, progress in the molecular genetics of Arabidopsis (the second area) may provide a more rapid route to understanding the control of flowering in cereals for several reasons: its small genome (1/4 that of rice); the likelihood of extensive homoeology with cereals, in spite of differences in codon usage between monocots and dicots; the existence of a wide range of flowering-time mutants; and the control of floral induction by a similar range of environmental factors including photoperiod and low temperature. It is likely that the MCDK (Martinez-Zapater, Coupland, Dean and Koornneef, 1994. In: Meyerowitz EM, Somerville CR. Arabidopsis. New York: Cold Spring Harbor Laboratory, 403–433) model, formulated to explain the genetic and environmental control of flowering in Arabidopsis, could be employed usefully in the formulation of experimental work on flowering in wheat and barley. This paper reviews these issues, paying particular attention to the significance of ‘earliness per se ’ loci and the ‘constitutive floral pathway’ for wheat and barley. Copyright 1998 Annals of Botany Company