• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of geneticsGeneticsCurrent IssueInformation for AuthorsEditorial BoardSubscribeSubmit a Manuscript
Genetics. May 2007; 176(1): 599–609.
PMCID: PMC1893030

The FLOWERING LOCUS T-Like Gene Family in Barley (Hordeum vulgare)

Abstract

The FLOWERING LOCUS T (FT) gene plays a central role in integrating flowering signals in Arabidopsis because its expression is regulated antagonistically by the photoperiod and vernalization pathways. FT belongs to a family of six genes characterized by a phosphatidylethanolamine-binding protein (PEBP) domain. In rice (Oryza sativa), 19 PEBP genes were previously described, 13 of which are FT-like genes. Five FT-like genes were found in barley (Hordeum vulgare). HvFT1, HvFT2, HvFT3, and HvFT4 were highly homologous to OsFTL2 (the Hd3a QTL), OsFTL1, OsFTL10, and OsFTL12, respectively, and this relationship was supported by comparative mapping. No rice equivalent was found for HvFT5. HvFT1 was highly expressed under long-day (inductive) conditions at the time of the morphological switch of the shoot apex from vegetative to reproductive growth. HvFT2 and HvFT4 were expressed later in development. HvFT1 was therefore identified as the main barley FT-like gene involved in the switch to flowering. Mapping of HvFT genes suggests that they provide important sources of flowering-time variation in barley. HvFTI was a candidate for VRN-H3, a dominant mutation giving precocious flowering, while HvFT3 was a candidate for Ppd-H2, a major QTL affecting flowering time in short days.

THE timing of flowering during the year is an important adaptive trait throughout the angiosperms. Correct flowering ensures the greatest chance of pollination, seed set, and dispersal, and therefore reproduction of the species. Flowering is regulated by environmental and internal cues and the genetic basis of this control is best understood in Arabidopsis where the photoperiod, vernalization, giberellic acid, and autonomous pathways have been defined (recently reviewed by Boss et al. 2004; Jack 2004; Bäurle and Dean 2006). The pathways' major point of convergence are genes called pathway integrators, which in Arabidopsis are FLOWERING LOCUS T (FT), SUPPRESSOR OF OVEREXPRESSION OF CO 1 (SOC1) [also called AGAMOUS-LIKE 20 (AGL20)], and, to a lesser extent, LEAFY (LFY) (Boss et al. 2004; Jack 2004; Moon et al. 2005; Bäurle and Dean 2006).

Each pathway does not have the same influence on each pathway integrator. In the case of the photoperiod pathway, which is our primary interest, FT is predominant. It was first identified in Arabidopsis (Kardailsky et al. 1999; Kobayashi et al. 1999) and was shown to be a direct target of the nuclear protein CONSTANS (CO) (Samach et al. 2000; Wigge et al. 2005). CO transcription is regulated by the circadian clock and peaks ~16 hr after dawn. This peak of expression has to correspond to a period of exposure to light for the CO protein to be stable and to induce FT expression, and by this process FT expression is restricted to long days (Suarez-Lopez et al. 2001; Searle and Coupland 2004; Valverde et al. 2004). Although CO was also shown to induce SOC1 expression (Samach et al. 2000), recent data show that this is through the action of FT (Yoo et al. 2005). FT expression can be detected in the leaves, mainly in the vascular tissues (An et al. 2004), but also at the apical meristem where the FT protein interacts with FD, a bZIP transcription factor, to promote flowering (Abe et al. 2005; Wigge et al. 2005). It has been suggested that the FT mRNA itself moves from the leaves, where photoperiod is perceived, to the apex where flowering is promoted (Huang et al. 2005).

The genetic basis of photoperiod response has also been studied extensively in rice (Oryza sativa), a monocot in which flowering is promoted by short days. Despite the evolutionary separation from Arabidopsis and the contrasting flowering response, photoperiod pathway genes are well conserved, with OsGI, Hd1, and Hd3a being orthologous to Arabidopsis GIGANTEA (GI), CO, and FT, respectively (Yano et al. 2000; Hayama et al. 2002; Kojima et al. 2002). It was shown that Hd1 represses Hd3a (FT) expression in long days (Hayama and Coupland 2004) but promotes Hd3a (FT) expression in short days, leading to flowering. These results show that variation in the COFT interplay is at the center of the long-day/short-day difference, but in both situations the induction of FT expression consistently promotes flowering.

The above data make FT of central interest to the photoperiodic regulation of flowering. However, interpreting the exact role of FT is complicated by variation in the structure of the FT family in different plants. In Arabidopsis, FT encodes a protein similar to a phosphatidylethanolamine-binding protein (PEBP), such as Raf kinase inhibitor (RKIP) from mammals (Kardailsky et al. 1999). It is a member of a small gene family, which includes five other genes: TERMINAL FLOWER 1 (TFL1), TWIN SISTER OF FT (TSF), ARABIDOPSIS THALIANA CENTRORADIALIS (ATC), BROTHER OF FT AND TFL1 (BFT), and MOTHER OF FT AND TFL1 (MFT) (Kobayashi et al. 1999). BFT has not been implicated in flowering (Yoo et al. 2004), but constitutive expression of FT, TSF, and, to a lesser extent, MFT accelerates flowering (Kobayashi et al. 1999; Yoo et al. 2004; Yamaguchi et al. 2005). Constitutive expression of TFL1 or ATC delays flowering (Mimida et al. 2001). However, FT and TFL1 proteins share ~59% amino acid identity and it was shown that swapping a single amino acid in the PEPB domain is sufficient to convert TFL1 to FT function and vice versa (Hanzawa et al. 2005).

Thirteen FT-like sequences have been found in the rice genome. These have been designated OsFTL1OsFTL13 with Hd3a corresponding to OsFTL2 (Izawa et al. 2002; Chardon and Damerval 2005; Zhang et al. 2005). Chardon and Damerval (2005) show that the higher number in rice can be attributed, in part, to duplication of chromosome regions within the rice genome. These duplications are thought to predate the divergence of the major grass lineages (Paterson et al. 2003; Salse et al. 2004) and so eight FT-like genes were suggested to have been present in the grass ancestral genome. At least three FT-like genes (OsFTL1, OsFTL2, and OsFTL3) are known to be active and capable of promoting flowering in rice (Izawa et al. 2002).

Barley (Hordeum vulgare) is more closely related to rice than to Arabidopsis but resembles the latter in photoperiod response. Several photoperiod pathway gene homologs have been identified in barley, such as HvGI, HvCO1, and HvCO2 (Griffiths et al. 2003; Dunford et al. 2005), and four FT-like EST consensus sequences were identified in barley through a database search (Chardon and Damerval 2005). Because of the importance of FT in flowering, it is important to define the structure of the family in temperate grasses. This article discusses the cloning, sequencing, and gene expression analysis of FT-like genes in barley and their phylogenetic relationship to rice and Arabidopsis FT-like genes. Our emphasis is on characterizing the genes most closely related to FT and likely to have a role as pathway integrators activated by photoperiod.

MATERIALS AND METHODS

Searches of The Institute for Genomic Research rice pseudomolecules and the Barley Gene Index for FT-like genes:

All the databases used for the searches are available at The Institute for Genomic Research (TIGR; http://www.tigr.org/) (Yuan et al. 2005) and at Gramene (http://www.gramene.org/) (Jaiswal et al. 2006). Starting with the Arabidopsis FT protein sequence FT_ARATH (Q9SXZ2), a BLASTP search was carried out against the predicted proteins from the TIGR rice pseudomolecules release 4. In addition, a TBLASTN search was carried out against all rice bacterial artificial chromosome (BAC) and P1-artificial chromosome (PAC) sequences in GenBank (http://www.ncbi.nlm.nih.gov/) to search for genes not present or not correctly annotated in the TIGR gene models.

A multiple sequence alignment of the TIGR gene models was made using CLUSTAL W (http://www.ebi.ac.uk/) (Thompson et al. 1994) against the conserved PEBP domain (PF01161). New gene predictions were made using FGENESH+ and PROT_MAP (http://sun1.softberry.com) for FT-like genes showing incorrect alignment within the PEBP domain and those not predicted by TIGR (supplemental Table S1 at http://www.genetics.org/supplemental/).

Starting with the rice OsFTL2 (Os06g06320) encoded protein (11976.m05358), a TBLASTN search was carried out against the Barley TIGR Unique Gene Indices release 9.0 (Lee et al. 2005). Gene index mapping information available from TIGR was used to align the barley sequences to the rice FT-like genes. Representative clones were selected from the ESTs used to construct the TIGR Gene Indices for the screening of a barley BAC library.

BAC library analysis:

High-density filters of a Morex barley BAC library with approximately sixfold genome coverage were purchased from Clemson University Genomics Institute (CUGI; http://www.genome.clemson.edu). Three EST clones containing full-length cDNAs were also obtained. HVSMEh0101D16 (BE602964) corresponding to the FT-like unigene sets TC143893 and TC143873 was obtained from CUGI, while baak20f16 (BJ448552) for TC151142 and bahl1n08 (AV937451) for TC152179 were provided by the Barley Germplasm Center, Okayama University (Kurashiki, Japan). Hybridization probes were amplified from the three EST clones using sequence-specific primers (HVSMEh0101D16: forward—5′-gacgtggtggacccgttc-3′ and reverse—5′-cagtcggtggatcccgag-3′; baak20f16: forward—5′-ccgttccattaaggatagcc-3′ and reverse—5′-ccatccggtggataccag-3′; bahl1n08: forward—5′-ctcagtgcctctaactgtgatg-3′ and reverse—5′-cagctgctggtacagaac-3′). An OsFTL2-specific probe was amplified from Nipponbare rice genomic DNA using primers designed for the OsFTL2 sequence as described in Turner et al. (2005). After hybridization screens with all four probes using the Church and Gilbert (1984) method, five barley BACs, each containing one of five FT-like genes, were investigated in detail. Primer pairs (supplemental Table S2 at http://www.genetics.org/supplemental/) amplifying overlapping segments of the all FT-like genes were designed from the barley EST sequences used as probes. Two BACs (236M13 and 440G4) were subcloned using the TOPO Shotgun subcloning kit (Invitrogen, San Diego) to obtain 5′- and 3′-end sequences. Additionally, coding regions and intervening introns for HvFT1, HvFT3, and HvFT4 were amplified from the barley cultivars Igri and Triumph. The absence of intron 3 in HvFT1 was investigated in wild barley (H. spontaneum) accessions and in the wheat cultivar Chinese Spring using the following primers: forward—5′-gttggtgacagatatccgg-3′ and reverse—5′-ccctggtgttgaagttctgg-3′.

Phylogenetic analysis of the PEBP domain of Arabidopsis, rice, and barley FT-like genes:

Multiple sequence alignment with CLUSTAL X of the PEBP domain (Pfam PF01161) for the two Arabidopsis proteins AtFT (FT_ARATH, Q9SXZ2) and AtTSF (TSF_ARATH, Q9S7R5), 13 rice FT-like proteins, and five barley FT proteins, together with the outgroup sequence OsMFTL1 (encoded by the gene Os06g30370), was used to generate input files for phylogenetic analysis. A tree was constructed using PHYLIP 3.5 (Felsenstein 1993). Bootstraps with 100 replicates were performed to assess node support. Multiple sequence alignment data were read in using SEQBOOT to produce multiple data sets for bootstrap resampling to, in turn, produce 100 data sets. PROTDIST (Dayhoff PAM matrix) using 100 data sets computed a distance measure using maximum-likelihood estimates. NEIGHBOR was used to produce an unrooted tree using the neighbor-joining method with one outgroup species (OsMFTL1). CONSENSE was used to draw the tree by majority rule, including bootstrap values. The tree was viewed using ATV: A Tree Viewer (Zmasek and Eddy 2001).

Genetic mapping:

A probe specific for each barley FT-like gene was hybridized to wheat/barley telosomic addition lines (Islam 1983) to assign the barley genes to chromosome arms. Subsequently, HvFT1, HvFT2, HvFT4, and HvFT5 were mapped in an F6 RI population from an Igri × Dairokkaku cross. All four were mapped using single-strand conformation polymorphism (Martins-Lopes et al. 2001) after amplification using the primers described in supplemental Table S2 at http://www.genetics.org/supplemental/. HvFT3 was mapped in an Igri × Triumph doubled haploid (DH) population (Laurie et al. 1995) as a presence/absence polymorphism using the cDNA probe used to screen the BAC library.

Collinearity of the HvFT1, HvFT2, HvFT3, and HvFT4 regions with rice was investigated by identifying additional closely linked genes. Sequences of rice BAC clones carrying the respective homologous OsFTL genes were used for BLASTN searches to identify corresponding barley ESTs, which were used to design PCR primers (supplemental Table S3 at http://www.genetics.org/supplemental/). These primers were used to amplify from the parents of the two mapping populations described above and from the BAC clones containing the respective HvFT genes. PCR reactions were as follows: in a total reaction volume of 20 μl, 50 ng of DNA, 0.2 μm of primers, 0.2 μm of dNTPS, 1× Taq polymerase buffer (Roche), and 0.4 unit of Taq polymerase (Roche). The PCR conditions were an initial denaturation at 94° for 2 min, followed by 40 cycles of denaturation at 94° for 30 sec, annealing at 55° for 30 sec, and extension at 72° for 1 min, followed by a final extension of 5 min at 72°. Single-strand conformation polymorphism (Martins-Lopes et al. 2001) was used to genetically map polymorphic markers.

Gene expression analysis:

Gene-specific primers (supplemental Table S2 at http://www.genetics.org/supplemental/) for each HvFT gene were used to assay expression in mature embryos (after stratification) and in plants grown for 7, 14, 21, and 28 days in long-day (LD; 16 hr light) and short-day (SD; 8 hr light) conditions (136 μmol m−2 sec−1, 22° during the day, 18° during the night). The genotype used was Triumph into which a functional Ppd-H1 allele has been introgressed as described in Turner et al. (2005). This isogenic line [Triumph(Ppd-H1)] is therefore responsive to photoperiod but lacks any vernalization requirement. Reports in Arabidopsis suggested that FT expression was relatively high 4 hr after the start of the dark phase (Kardailsky et al. 1999; Kobayashi et al. 1999, Suarez-Lopez et al. 2001). Plants were therefore sampled 4 hr after dusk with each sample comprising six plants. RNA was extracted, cDNA was synthesized, and samples were processed using an Opticon 2 real time PCR instrument (http://www.mjr.com) as described in Turner et al. (2005).

RESULTS

Database searches for FT-like genes in rice and barley:

A BLASTP search of peptides (TIGRv4 gene models) using FT_ARATH identified 12 FT-like genes from rice corresponding to all the FT-like genes described by Chardon and Damerval (2005) except OsFTL5. A TBLASTN search of all rice BAC and PAC sequences found an additional FT-like gene on BAC AP004124 corresponding to OsFTL5. This gene is not annotated as a predicted gene by the TIGR annotation project (Yuan et al. 2005). The chromosome position and BAC location of the genes was used to verify that the 13 FT-like genes identified by Chardon and Damerval (2005) corresponded to the correct TIGR loci (Table 1).

TABLE 1
Rice and barley FT-like genes

The protein sequences for all the rice FT-like genes except OsFTL5 were downloaded from TIGR and a multiple alignment was performed using ClustalW. This showed that predicted protein sequences for OsFTL4, -8, -9, and -10 did not align with the PEBP domain. New protein predictions were carried out for these genes and for OsFTL5 (Figure 1; supplemental Table S1 at http://www.genetics.org/supplemental/). The OsFTL9 protein from Nipponbare (subspecies japonica) contained a stop codon in the PEBP domain and is predicted to be nonfunctional. The OsFTL9 protein from 93-11 (subspecies indica) was intact (Figure 1).

Figure 1.
ClustalW multiple alignment of the complete protein sequences of Arabidopsis, rice, and barley FT family proteins. OsMFTL1, used for construction of the phylogenetic tree (Figure 2A), is also included. The PEBP domain boundaries are marked by horizontal ...

A TBLASTN search of the barley TIGR Gene Indices database using OsFTL2 (Hd3a) as a template retrieved four barley tentative consensus (TC) sequences as described by Chardon and Damerval (2005) (Table 1).

BAC library screens:

No barley ESTs closely matched the rice OsFTL2 (Hd3a) sequence. To identify this gene, a nucleotide probe derived from OsFTL2 was used to screen a Morex barley BAC library. Subclones from BAC clone 440G4 were identified as containing a Hd3a homolog and were sequenced, revealing an FT-like gene highly homologous to OsFTL2 (Hd3a) and FT. This gene was designated HvFT1. The BAC library was also screened with probes specific to each barley EST contig (Table 1). A total of 32 positive BACs were obtained and, after fingerprinting and hybridizing with the same probes, they were classified into four groups. Two distinct classes corresponded to HvFT2 and HvFT4 while cross-hybridization suggested that two other genes (HvFT3 and HvFT5) were closely related and might be the result of a recent duplication. Full gene sequence was obtained for each of these genes from a representative BAC clone (Table 1; Figure 1).

Phylogenetic analysis of the PEBP domain of Arabidopsis, rice, and barley FT-like genes:

The evolutionary relationship between the FT-like genes in Arabidopsis, rice, and barley was investigated by constructing a phylogenetic tree using neighbor-joining of the PEBP domain protein sequences (Figure 2A). Protein sequence homology measured as the percentage of identical protein residues is shown in supplemental Table S4 at http://www.genetics.org/supplemental/. The FT family fell into three groups. The seven genes in group 1 were the most FT-like and comprised AtFT, AtTSF, OsFTL1-3, and HvFT1-2 (Figure 2A). Rice and barley proteins had 71–73% identity with AtFT and there is 79–89% identity between rice and barley proteins. HvFT1 was most similar to OsFTL2 while HvFT2 was highly homologous to OsFTL1.

Figure 2.
Phylogenetic relationships of FT-like proteins from Arabidopsis, rice, and barley. (A) Phylogenetic tree of PEBP domain protein sequences. Major groups are marked 1–3. Bootstrap support values are shown at each node. (B) Exon/intron structure ...

Group 2 proteins (58–65% identity with AtFT) included HvFT3, HvFT4, and HvFT5. HvFT4 was very similar to OsFTL12, while HvFT3 and HvFT5 were highly homologous to each other and are related to the pair of rice proteins OsFTL9 and OsFTL10. Group 3 contained rice sequences with 63–71% homology with AtFT but no barley proteins. Chardon and Damerval (2005) also found no barley ESTs homologous to these rice genes. If present in barley, as seems likely, they were too diverged in sequence to be detected in the BAC library hybridization screen.

Hanzawa et al. (2005) showed that the tyrosine (Y) amino acid at position 85 in AtFT is critical to FT function and that replacing this with a histidine (H) was sufficient to convert FT to TFL1 in terms of function. The reciprocal substitution converted TFL1 to FT. All the FT-like genes in Figure 1 have the critical Y consistent with being true FT-like genes and likely to have activator-type roles in flowering. A barley TFL1 homolog (DQ539338) has the expected H residue at this position.

More recently, Ahn et al. (2006) identified a 14-amino-acid stretch in which 11 amino acids were invariant in all FT-like proteins that they analyzed compared to 4 in all TFL1-like proteins. A single residue unambiguously distinguished between FT and TFL1 homologs: Gln140 in FT and Asp144 in TFL1. Group 1 proteins show the same invariance (boxed in Figure 1), further suggesting that they are the most FT-like. Group 2 and group 3 proteins have some variation in this stretch. HvFT5 has a histidine in place of the Gln140 and HvFT3; HvFT4 and HVFT5 have at least 3 amino acids varying of the 11 amino acids characterized as invariant. Furthermore, Ahn et al. (2006) also observed an essentially invariant triad starting at residue 150 with a leucine or isoleucine, followed by a tyrosine and an asparagine. This triad is present only in the group 1 and in some of the group 3 proteins (boxed in Figure 1). Taken together, these observations confirm that proteins from group 1 are the most FT-like, while group 2 and group 3 proteins are more diverged.

The exon/intron structure of the barley FT-like genes:

In Arabidopsis, FT and TSF have four exons and three introns (Figure 2B). The first and fourth exons are the largest (264 and 463 bp, respectively), while exons 2 and 3 are smaller (61 and 40 bp, respectively). Intron 1 and 2 are similar in size (815 and 713 bp, respectively), but intron 3 is smaller (205 bp). This structure is well conserved across all the FT-like genes found in the rice genome, although some variation is found in the relative sizes of the introns. In barley, HvFT2, HvFT3, HvFT4, and HvFT5 also have this structure, but HvFT1 is distinct in lacking the third intron, perfectly merging exons 3 and 4 together (Figure 2B). The sequence was obtained from a BAC clone derived from the spring barley cultivar Morex. The loss of the intron was confirmed in cultivars Igri and Triumph, in H. spontaneum accessions, and in the three group 7 chromosome wheat nullisomic/tetrasomic lines derived from the variety Chinese Spring, showing that the intron is absent in all three wheat genomes (supplemental Figure S1 at http://www.genetics.org/supplemental/).

Expression profiles under LD and SD in a Ppd-H1 genetic background:

Gene expression was studied in Triumph(Ppd-H1) plants, which do not require vernalization but are highly responsive to a long-day photoperiod (Turner et al. 2005). Expression levels of the five HvFT genes were compared in plants grown under LD (16 hr light) or SD (8 hr light) conditions. For HvFT1, HvFT2, and HvFT4, no or extremely low expression was detected under SD conditions at all time points (Figure 3a). In LD conditions, HvFT1 was rapidly induced and was detected after 1 week, while HvFT2 and HvFT4 expression was significantly induced after 3–4 weeks (Figure 3a). Dissection of developing apices showed that the appearance of the double ridge stage, the first visible sign of the switch from vegetative to floral development (Kirby and Appleyard 1981), occurred during the second week under LD and during the fourth week under SD conditions (Figure 3b). Therefore, HvFT1 was the only FT-like gene highly expressed at the transitional phase under LD conditions.

Figure 3.
Expression of HvFT genes under short-day and long-day conditions. (a) Levels of gene expression in arbitrary units normalized against 18s rRNA. (b) Timing of the transition of the developing apex from vegetative to reproductive growth (extension of the ...

HvFT3 was unusual in being strongly expressed under SD (noninductive) conditions from week 1, but being very weakly expressed under LD (inductive) conditions (Figure 3a). Barley plants will flower in SD conditions but the apical transition in SDs occurred while HvFT1 expression remained very low, suggesting that this is not due to a low-level induction of genes normally involved in the long-day response. Possibly a second mechanism using a different FT gene is involved. HvFT3 may be a candidate for this, but HvFT3 expression increased well before the apical transition in SDs, suggesting that HvFT3 expression is not sufficient to induce the transition. HvFT4 was weakly expressed during the transitional phase in SD conditions and may play a role. HvFT3 was also unusual in being expressed in the embryo after stratification, but in this case the transcript was unspliced (supplemental Figure S2a at http://www.genetics.org/supplemental/) and is predicted to give a nonfunctional protein.

Expression of an unspliced HvFT5 transcript was detected at all time points in LD and SD conditions (supplemental Figure S2b at http://www.genetics.org/supplemental/). The predicted protein had a stop codon at residue 69 and is likely to be nonfunctional. The correctly spliced form was detected after 4 weeks in SDs, but the expression level was very low.

Comparative mapping of the five barley FT-like genes:

Phylogenetic analysis of the PEBP domain of barley and rice FT-like protein sequences showed that the five barley genes could each be associated with one or two different rice genes (Figure 2A). Comparative mapping of barley with rice was used to provide additional information on the relationships among genes. First, hybridizations of probes specific to each of the five HvFT genes to wheat/barley telosomic addition lines were used to assign individual barley genes to chromosome arms. HvFT genes were then genetically mapped (Figure 4). For additional comparative mapping, rice genes present on the same BAC clones as the putative homologous OsFT-like genes were used for BLASTN searches (http://www.ncbi.nlm.nih.gov/) to identify corresponding barley ESTs. Matching barley ESTs were used to develop markers for genetic and physical mapping in barley.

Figure 4.
Genetic map positions of barley HvFT genes and their relationships to rice. Solid lines show the mapped segments in relation to the approximate size of complete barley linkage groups (H). Barley BACs are shown in boldface type. Shaded lines show homologous ...

HvFT1 was mapped to the short arm of chromosome 7H, between microsatellite markers AF022725 and Bmac31. The HvFT1 protein was highly homologous to the OsFTL2 and OsFTL3 proteins. These two rice genes are ~12 kb apart and are likely to be the result of a recent duplication. Among the 39 other genes predicted on the same BAC (AP005828), Os06g06410 was highly homologous to barley EST Ebro08_SQ010_A17. This sequence was mapped in barley by SSCP to the short arm of chromosome 7H, 0.3 cM distal to HvFT1. This shows that the region carrying HvFT1 is orthologous to the OsFTL2/3 region in rice. There was no evidence of a tandem duplication in barley from sequencing or from Southern hybridizations, which showed single-band profiles.

HvFT2 was mapped to the short arm of chromosome 3H, between microsatellite marker Bmac67 and STS marker MWG985. HvFT2 was most similar to OsFTL1 (Os01g11940). A rice BAC (AP002745) containing OsFTL1 also contained two genes (Os01g11946 and Os01g11952) with matching barley ESTs (Bags35l04 and HVSMEg0012L06) that were present on the HvFT2 barley BAC 236M13.

HvFT4 was mapped to the short arm of chromosome 2H, proximal to marker cMWG663. HvFT4 was highly homologous to OsFTL12 (Os06g35940), located on BAC AP003682 from rice chromosome 6. Barley EST HVSMEi0006N22 was homologous to Os06g35910, present on the same rice BAC. HVSMEi0006N22 was present on barley BAC 641D22, which also carried HvFT4, confirming that HvFT4 is the ortholog of OsFTL12. OsFLT12 and -13 are likely to derive from duplication within the rice genome. No equivalent duplication was found in barley, and Southern hybridization using an HvFT4 probe yielded a single band.

HvFT3 was mapped to the long arm of chromosome 1H, cosegregating with marker Xpsr162. The HvFT3 protein was highly homologous to OsFTL10 encoded by the rice gene Os05g44180 present on a rice chromosome 5 BAC (AC130603). Two rice genes also present on this BAC (Os05g44050 and Os05g44170) were homologous to the barley ESTs BES1824107g14 and HO07N13, respectively. BES1824107g14 was mapped in barley by SSCP and cosegregated with HvFT3, while HO07N13 was both mapped by SSCP, co-segregating with HvFT3, and found on barley BAC 347D22, which also carried HvFT3. HvFT3 was therefore confirmed as the ortholog of OsFTL10. HvFT5 was also highly homologous to OsFTL10, but was mapped to the long arm of chromosome 4H, cosegregating with marker scsnp20989. Markers in the region of HvFT5 have homologous sequences at the distal ends of rice chromosome 3S and 10L, which do not contain any FT-like genes. The lack of an equivalent rice gene to HvFT5 and the high sequence homology of HvFT3 and HvFT5 suggest that the latter is the result of a duplication that occurred after the divergence of barley and rice. Therefore, while OsFTL9, OsFTL10, HvFT3, and HvFT5 were related (Figure 2A), they are likely to derive from independent duplications. Absence of the OsFTL9 equivalent from barley was supported by hybridizing OsFTL9 and OsFTL10 probes amplified from rice to wheat/barley telosomic addition lines. OsFTL10 showed strong hybridization to both wheat and barley while OsFTL9 hybridized only weakly to the same fragments as OsFTL10 and detected no additional band.

Associations of HvFT genes with flowering-time QTL:

Three of the HvFT genes are in regions previously shown to contain flowering-time QTL in an Igri × Triumph cross (Laurie et al. 1995). HvFT4 mapped to the region of barley chromosome 2 where Laurie et al. (1995) mapped eps2S, and HvFT1 was in the same region as eps7S. Both QTL are earliness factors with no obvious relationship to photoperiod response (Laurie et al. 1995). No direct evidence was found for these being the underlying genes as there was no polymorphism in the HvFT4 or HvFT1 coding regions between Igri and Triumph. However, this does not exclude the possibility of differences in expression. An Hd3a-like FT gene was also associated with a QTL for early flowering that was found on chromosome 7 in Lolium perenne in a collinear position to OsFTL2 in rice (Armstead et al. 2002). Interestingly, the Igri × Dairokakku population used to map HvFT1 also segregated for VRN-H3, a vernalization locus previously mapped to 1HL (Takahashi and Yasuda 1971). However, VRN-H3 cosegregated with HvFT1 on chromosome 7H in our population, consistent with recent findings by Yan et al. (2006) who showed that spring habit mutations in Vrn-B3 lines of wheat and Vrn-H3 lines of barley are due to altered expression of HvFT1 (in our nomenclature). No sequence polymorphism was observed in the coding regions of HvFT1 between Igri and Dairokakku, but, as found by Yan et al. (2006), nine SNP and three indels were found in the 648 bases of the promoter region upstream of the start codon, three SNPs were present in the first intron, and a polymorphic SSR was observed in the second intron (supplemental Figure S3 at http://www.genetics.org/supplemental/).

HvFT3 was also interesting because it was closely associated with Photoperiod-H2 (Ppd-H2). This major QTL affected flowering time in a short-day glasshouse experiment (10 hr light) and in an autumn-sown field experiment but was not detectable in long days (Laurie et al. 1995), consistent with the observed expression pattern of HvFT3 (Figure 3). In short days, Igri contributed the late-flowering allele. The Triumph sequence for HvFT3 was identical to Morex but no fragment could be amplified from Igri. A presence/absence result from Southern hybridizations showed the gene to be at least partly deleted from Igri. Reanalysis of the SD glasshouse experiment suggested that at least one line (DH48, Figure 5) showed recombination between HvFT3 and Ppd-H2. However, analysis using JoinMap 4.0 (Plant Research International B.V.) showed additional QTL to be present. By using the Ppd-H2 region as a cofactor, additional significant QTL were localized on 2H, 5H, and 6H and the additive effects of these QTL could account for the range of flowering times in DH lines 48, 40, and 61 (Figure 5; supplemental Figure S4 at http://www.genetics.org/supplemental/). This ambiguity means that HvFT3 is a sufficiently good candidate for Ppd-H2 to warrant further study.

Figure 5.
Flowering times (days to awn emergence from the flag leaf) of DH lines from an Igri × Triumph population showing the association of early flowering with the Triumph allele of HvFT3. The effect of additional QTL is shown in supplemental Figure ...

To test whether Igri was unusual in having a deletion of HvFT3, we used the presence/absence PCR polymorphism to analyze a sample of 60 spring and 40 winter barley cultivars from Europe (supplemental Table S5 at http://www.genetics.org/supplemental/). The Triumph allele was prevalent in spring types (46/60, 78%) and the Igri allele in winter types (36/40, 90%), and this association was highly significant (χ2 1, d.f. 43.7, P < 0.001). Both alleles could be found in two-row and six-row types. The prevalence of the deletion in winter types suggests that this may have been selected to enhance the suppression of flowering in overwintering plants.

DISCUSSION

Our main interest was the identification of barley genes most similar to Arabidopsis FT and hence most likely to be significant as floral pathway integrators. The barley genes that we identified correspond to rice genes in groups 1 and 2 of Figure 2. Group 3 genes may exist in barley, but no ESTs have been found and no clones were identified in our library screens, probably because the nucleotide sequence is too diverged.

Differences in gene number between rice and barley are primarily attributable to differences in the fate of duplicated genes. HvFT1 corresponds to OsFLT2 and -3, which are likely to be a recent duplication. OsFTL9-10 and OsFLT12-13 are pairs resulting from duplications within the rice genome (Paterson et al. 2003; Salse et al. 2004), but for the former we detected only an equivalent of OsFLT10 (HvFT3) and for the latter only the equivalent of OsFLT12 (HvFT4), suggesting that two genes have been lost from barley. HvFT3 and HvFT5 are likely to derive from a more recent duplication in barley. HvFT2 corresponds to a single gene in rice, OsFTL1.

In rice, OsFTL2 was identified by positional cloning of a QTL for flowering time (Hd3a), showing significant natural variation at this locus (Kojima et al. 2002). The significance of this gene was confirmed by experiments on the effect of night break on the expression of rice FT-like genes. Only OsFTL2 expression was affected by night break, pointing to a key role for this gene in photoperiod response (Ishikawa et al. 2005). In barley, HvFT1 is highly homologous to OsFTL2 (89% in the PEBP domain) and maps to a collinear position on chromosome 7H. HvFT1 was the gene most rapidly upregulated by long days and the only HvFT-like gene to be upregulated at the time of apical transition. Expression of this gene was also found to be significantly reduced in the barley ppd-H1 mutant, which is late flowering in long days (Turner et al. 2005). Furthermore, Yan et al. (2006) have shown that allelic variation in HvFT1 is associated with large differences in flowering time. These data suggest that this gene is a prime candidate for a floral pathway integrator in barley, but this does not exclude roles for the other FT-like genes.

An unusual feature of HvFT1 was the absence of the third intron present in AtFT, all rice FT-like genes, and all other HvFTs. Reverse transcription of spliced mRNAs followed by homologous recombination of the cDNA with the genomic copy of the gene has been suggested as a mechanism for precise intron loss (Roy and Gilbert 2006), and this process has been shown to occur in unicellular eukaryotes (Bon et al. 2003; Mourier and Jeffares 2003; Sverdlov et al. 2004; Roy and Gilbert 2005). The loss of the intron was found in several barley cultivars, wild barley accessions, and wheat, suggesting that it may be a general feature of temperate grasses.

A surprising finding was the potential role of an FT gene in the control of flowering time under noninductive (SD) conditions. Under SD conditions, HvFT3 was expressed from the first week at a higher level than under LD conditions, while HvFT4 was very weakly expressed after 3 and 4 weeks, and HvFT1 and HvFT2 were not expressed at all. The switch at the apex from vegetative to floral meristem occurred between the third and the fourth weeks, suggesting that HvFT4 could be involved in the promotion of flowering under SD or that an additional pathway is in place to promote flowering under SD in barley. A priority for understanding flowering in SDs is to resolve the relationship between HvFT3 and Ppd-H2, which can be done by further mapping and tests of gene function using transgenic approaches.

Barley and rice diverged from an ancestral grass ~60 MYA (Gale and Devos 1998) and have contrasting photoperiodic responses. This is achieved using photoperiod pathway genes that are well conserved with more distantly related plants like Arabidopsis. Furthermore, the photoperiod response in barley and rice has been modified considerably during domestication. A challenge for cereal biology is to reconcile this diversity with the conservation of the underlying pathway. To do this, it is important to understand the pathway components and to develop models that accommodate variation in the structure of gene families.

Hayama and Coupland (2004) showed that although both CO and FT are conserved between rice and Arabidopsis, their relationship is different. In Arabidopsis, CO promotes FT expression under LD conditions only, while in rice, CO represses FT under LDs and promotes it under SDs. Barley is a LD plant, like Arabidopsis, but is phylogenetically closer to rice, a SD plant. Barley differs from rice and Arabidopsis in having two CO-like genes (HvCO1 and HvCO2) of which the former is unusual in having lost key residues in a normally highly conserved zinc-finger domain (B-box 2) (Griffiths et al. 2003). Characterization of the roles and interactions of the various CO and FT genes in barley is now feasible because of the increased understanding of the gene families and the availability of new resources for functional analysis, including efficient transformation methods (Travella et al. 2005) and a TILLING population (Caldwell et al. 2004).

Acknowledgments

The authors are grateful to Donal O'Sullivan (National Institute of Agricultural Botany, Cambridge, UK) for kindly providing DNA samples of the barley GEDIFLUX collection tested for the HvFT3 allele. We also thank S. Yasuda (Research Institute for Bioresources, Okayama University, Kurashiki, Japan) for providing the barley Vrn-H3 genetic stock. This work was supported by the United Kingdom Biotechnology and Biological Sciences Research Council through grant 208/D19952 and by a grant-in-aid to the John Innes Centre. J.H. was supported by a fellowship from the Daphne Jackson Trust funded by The Gatsby Charitable Foundation.

Notes

Sequence data from this article have been deposited with the EMBL/GenBank data libraries under accession nos. DQ297407, DQ411319, DQ411320, and EF012202.

References

  • Abe, M., Y. Kobayashi, S. Yamamoto, Y. Daimon, A. Yamaguchi et al., 2005. FD, a bZIP protein mediating signals from the floral pathway integrator FT at the shoot apex. Science 309: 1052–1056. [PubMed]
  • Ahn, J. H., D. Miller, V. J. Winter, M. J. Banfield, J. Hwan Lee et al., 2006. A divergent external loop confers antagonistic activity on floral regulators FT and TFL1. EMBO J. 25: 605–614. [PMC free article] [PubMed]
  • An, H. L., C. Roussot, P. Suarez-Lopez, L. Corbesler, C. Vincent et al., 2004. CONSTANS acts in the phloem to regulate a systemic signal that induces photoperiodic flowering of Arabidopsis. Development 131: 3615–3626. [PubMed]
  • Armstead, I. P., L. B. Turner, I. P. King, A. J. Cairns and M. O. Humphreys, 2002. Comparison and integration of genetic maps generated from F-2 and BC1-type mapping populations in perennial ryegrass. Plant Breed. 121: 501–507.
  • Bäurle, I., and C. Dean, 2006. The timing of developmental transitions in plants. Cell 125: 655–664. [PubMed]
  • Bon, E., S. Casaregola, G. Blandin, B. Llorente, C. Neuveglise et al., 2003. Molecular evolution of eukaryotic genomes: hemiascomycetous yeast spliceosomal introns. Nucleic Acids Res. 31: 1121–1135. [PMC free article] [PubMed]
  • Boss, P. K., R. M. Bastow, J. S. Mylne and C. Dean, 2004. Multiple pathways in the decision to flower, enabling, promoting and resetting. Plant Cell 16: S18–S31. [PMC free article] [PubMed]
  • Caldwell, D. G., N. McCallum, P. Shaw, G. J. Muehlbauer, D. F. Marshall et al., 2004. A structured mutant population for forward and reverse genetics in barley (Hordeum vulgare L.). Plant J. 40: 143–150. [PubMed]
  • Chardon, F., and C. Damerval, 2005. Phylogenomic analysis of the PEBP gene family in cereals. J. Mol. Evol. 61: 579–590. [PubMed]
  • Church, G. M., and W. Gilbert, 1984. Genomic sequencing. Proc. Natl. Acad. Sci. USA 81: 1991–1995. [PMC free article] [PubMed]
  • Dunford, R. P., S. Griffiths, V. Christodoulou and D. A. Laurie, 2005. Characterisation of a barley (Hordeum vulgare L.) homologue of the Arabidopsis flowering time regulator GIGANTEA. Theor. Appl. Genet. 110: 925–931. [PubMed]
  • Felsenstein, J., 1993. PHYLIP (Phylogeny Inference Package), version 3.5c. Department of Genetics, University of Washington, Seattle (http://evolution.genetics.washington.edu/phylip.html).
  • Gale, M. D., and K. M. Devos, 1998. Comparative genetics in the grasses. Proc. Natl. Acad. Sci. USA 95: 1971–1974. [PMC free article] [PubMed]
  • Griffiths, S., R. P. Dunford, G. Coupland and D. A. Laurie, 2003. The evolution of CONSTANS-like gene families in barley, rice, and Arabidopsis. Plant Physiol. 131: 1855–1867. [PMC free article] [PubMed]
  • Hanzawa, Y., T. Money and D. Bradley, 2005. A single amino acid converts a repressor to an activator of flowering. Proc. Natl. Acad. Sci. USA 102: 7748–7753. [PMC free article] [PubMed]
  • Hayama, R., and G. Coupland, 2004. The molecular basis of diversity in the photoperiodic flowering responses of Arabidopsis and rice. Plant Physiol. 135: 677–684. [PMC free article] [PubMed]
  • Hayama, R., T. Izawa and K. Shimamoto, 2002. Isolation of rice genes possibly involved in the photoperiodic control of flowering by a fluorescent differential display method. Plant Cell Physiol. 43: 494–504. [PubMed]
  • Huang, T., H. Bohlenius, S. Eriksson, F. Parcy and O. Nilsson, 2005. The mRNA of the Arabidopsis gene FT moves from leaf to shoot apex and induces flowering. Science 309: 1694–1696. [PubMed]
  • Ishikawa, R., S. Tamaki, S. Yokoi, N. Inagaki, T. Shinomura et al., 2005. Suppression of the floral activator Hd3a is the principal cause of the night break effect in rice. Plant Cell 17: 3326–3336. [PMC free article] [PubMed]
  • Islam, A., 1983. Ditelosomic additions of barley chromosomes to wheat, pp. 233–238 in Proceedings of the Sixth International Wheat Genetics Symposium, edited by S. Sakamoto. Plant Germ-plasm Institute, Faculty of Agriculture, Kyoto University, Kyoto, Japan.
  • Izawa, T., T. Oikawa, N. Sugiyama, T. Tanisaka, M. Yano et al., 2002. Phytochrome mediates the external light signal to repress FT orthologs in photoperiodic flowering of rice. Genes Dev. 16: 2006–2020. [PMC free article] [PubMed]
  • Jack, T., 2004. Molecular and genetic mechanisms of floral control. Plant Cell 16: S1–S17. [PMC free article] [PubMed]
  • Jaiswal, P., J. J. Ni, I. Yap, D. Ware, W. Spooner et al., 2006. Gramene: a bird's eye view of cereal genomes. Nucleic Acids Res. 34: D717–D723. [PMC free article] [PubMed]
  • Kardailsky, I., V. K. Shukla, J. H. Ahn, N. Dagenais, S. K. Christensen et al., 1999. Activation tagging of the floral inducer FT. Science 286: 1962–1965. [PubMed]
  • Kirby, E. J. M., and M. Appleyard, 1981. Cereal Development Guide. Cereal Unit, National Agricultural Centre, Stoneleih, Kenilworth, UK.
  • Kobayashi, Y., H. Kaya, K. Goto, M. Iwabuchi and T. Araki, 1999. A pair of related genes with antagonistic roles in mediating flowering signals. Science 286: 1960–1962. [PubMed]
  • Kojima, S., Y. Takahashi, Y. Kobayashi, L. Monna, T. Sasaki et al., 2002. Hd3a, a rice ortholog of the Arabidopsis FT gene, promotes transition to flowering downstream of Hd1 under short-day conditions. Plant Cell Physiol. 43: 1096–1105. [PubMed]
  • Laurie, D. A., N. Pratchett, J. H. Bezant and J. W. Snape, 1995. RFLP mapping of 5 major genes and 8 quantitative trait loci controlling flowering time in a winter × spring barley (Hordeum-Vulgare L) cross. Genome 38: 575–585. [PubMed]
  • Lee, Y., J. Tsai, S. Sunkara, S. Karamycheva, G. Pertea et al., 2005. The TIGR Gene Indices: clustering and assembling EST and known genes and integration with eukaryotic genomes. Nucleic Acids Res. 33: D71–D74. [PMC free article] [PubMed]
  • Martins-Lopes, P., H. Zhang and R. Koebner, 2001. Detection of single nucleotide mutations in wheat using single strand conformation polymorphism gels. Plant Mol. Biol. Rep. 19: 159–162.
  • Mimida, N., K. Goto, Y. Kobayashi, T. Araki, J. H. Ahn et al., 2001. Functional divergence of the TFL1-like gene family in Arabidopsis revealed by characterization of a novel homologue. Genes Cells 6: 327–336. [PubMed]
  • Moon, J., H. Lee, M. Kim and I. Lee, 2005. Analysis of flowering pathway integrators in Arabidopsis. Plant Cell Physiol. 46: 292–299. [PubMed]
  • Mourier, T., and D. C. Jeffares, 2003. Eukaryotic intron loss. Science 300: 1393. [PubMed]
  • Paterson, A. H., J. E. Bowers, D. G. Peterson, J. C. Estill and B. A. Chapman, 2003. Structure and evolution of cereal genomes. Curr. Opin. Genet. Dev. 13: 644–650. [PubMed]
  • Roy, S. W., and W. Gilbert, 2005. The pattern of intron loss. Proc. Natl. Acad. Sci. USA 102: 713–718. [PMC free article] [PubMed]
  • Roy, S. W., and W. Gilbert, 2006. The evolution of spliceosomal introns: patterns, puzzles and progress. Nat. Rev. Genet. 7: 211–221. [PubMed]
  • Salse, J., B. Piegu, R. Cooke and M. Delseny, 2004. New in silico insight into the synteny between rice (Oryza sativa L.) and maize (Zea mays L.) highlights reshuffling and identifies new duplications in the rice genome. Plant J. 38: 396–409. [PubMed]
  • Samach, A., H. Onouchi, S. E. Gold, G. S. Ditta, Z. Schwarz-Sommer et al., 2000. Distinct roles of CONSTANS target genes in reproductive development of Arabidopsis. Science 288: 1613–1616. [PubMed]
  • Searle, I., and G. Coupland, 2004. Induction of flowering by seasonal changes in photoperiod. EMBO J. 23: 1217–1222. [PMC free article] [PubMed]
  • Suarez-Lopez, P., K. Wheatley, F. Robson, H. Onouchi, F. Valverde et al., 2001. CONSTANS mediates between the circadian clock and the control of flowering in Arabidopsis. Nature 410: 1116–1120. [PubMed]
  • Sverdlov, A. V., V. N. Babenko, I. B. Rogozin and E. V. Koonin, 2004. Preferential loss and gain of introns in 3′ portions of genes suggests a reverse-transcription mechanism of intron insertion. Gene 338: 85–91. [PubMed]
  • Takahashi, R., and S. Yasuda, 1971. Genetics of earliness and growth habit in barley, pp. 388–408 in Proceedings of the 2nd International Barley Genetics Symposium, 6–11 July 1969, edited by R. A. Nilan. Washington State University Press, Pullman, WA.
  • Thompson, J. D., D. G. Higgins and T. J. Gibson, 1994. Clustal-W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 22: 4673–4680. [PMC free article] [PubMed]
  • Travella, S., S. M. Ross, J. Harden, C. Everett, J. W. Snape et al., 2005. A comparison of transgenic barley lines produced by particle bombardment and Agrobacterium-mediated techniques. Plant Cell Rep. 23: 780–789. [PubMed]
  • Turner, A., J. Beales, S. Faure, R. P. Dunford and D. A. Laurie, 2005. The pseudo-response regulator Ppd-H1 provides adaptation to photoperiod in barley. Science 310: 1031–1034. [PubMed]
  • Valverde, F., A. Mouradov, W. Soppe, D. Ravenscroft, A. Samach et al., 2004. Photoreceptor regulation of CONSTANS protein in photoperiodic flowering. Science 303: 1003–1006. [PubMed]
  • Wigge, P. A., M. C. Kim, K. E. Jaeger, W. Busch, M. Schmid et al., 2005. Integration of spatial and temporal information during floral induction in Arabidopsis. Science 309: 1056–1059. [PubMed]
  • Yamaguchi, A., Y. Kobayashi, K. Goto, M. Abe and T. Araki, 2005. TWIN SISTER OF FT (TSF) acts as a floral pathway integrator redundantly with FT. Plant Cell Physiol. 46: 1175–1189. [PubMed]
  • Yan, L., D. Fu, C. Li, A. Blechl, G. Tranquilli et al., 2006. The wheat and barley vernalization gene VRN3 is an orthologue of FT. Proc. Natl. Acad. Sci. USA 103: 19581–19586. [PMC free article] [PubMed]
  • Yano, M., Y. Katayose, M. Ashikari, U. Yamanouchi, L. Monna et al., 2000. Hd1, a major photoperiod sensitivity quantitative trait locus in rice, is closely related to the Arabidopsis flowering time gene CONSTANS. Plant Cell 12: 2473–2483. [PMC free article] [PubMed]
  • Yoo, S. Y., I. Kardailsky, J. S. Lee, D. Weigel and J. H. Ahn, 2004. Acceleration of flowering by overexpression of MFT (MOTHER OF FT AND TFL1). Mol. Cells 17: 95–101. [PubMed]
  • Yoo, S. K., K. S. Chung, J. Kim, J. H. Lee, S. M. Hong et al., 2005. CONSTANS activates SUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1 through FLOWERING LOCUS T to promote flowering in Arabidopsis. Plant Physiol. 139: 770–778. [PMC free article] [PubMed]
  • Yuan, Q. P., O. Y. Shu, A. H. Wang, W. Zhu, R. Maiti et al., 2005. The institute for genomic research Osa1 rice genome annotation database. Plant Physiol. 138: 17–26. [PMC free article] [PubMed]
  • Zhang, S. H., W. J. Hu, L. P. Wang, C. F. Lin, B. Cong et al., 2005. TFL1/CEN-like genes control intercalary meristem activity and phase transition in rice. Plant Sci. 168: 1393–1408.
  • Zmasek, C. M., and S. R. Eddy, 2001. ATV: display and manipulation of annotated phylogenetic trees. Bioinformatics 17: 383–384. [PubMed]

Articles from Genetics are provided here courtesy of Genetics Society of America
PubReader format: click here to try

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

  • EST
    EST
    Published EST sequences
  • Gene (nucleotide)
    Gene (nucleotide)
    Records in Gene identified from shared sequence links
  • MedGen
    MedGen
    Related information in MedGen
  • Nucleotide
    Nucleotide
    Published Nucleotide sequences
  • Protein
    Protein
    Published protein sequences
  • PubMed
    PubMed
    PubMed citations for these articles
  • Taxonomy
    Taxonomy
    Related taxonomy entry
  • Taxonomy Tree
    Taxonomy Tree

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...