MADS-box Research Articles

Transcriptome analysis of differentially expressed genes during anther development stages on male sterility and fertility in Cucumis melo L. line

The genic male sterility (MS) plays a major role in melon hybrids production, it could reduce the cost of pollination and increase the yield and quality. However, the molecular mechanism underlying genetic male sterility is yet poorly understood. The morphological differences of flower buds of melon were observed showed that the flower buds were tetrad when they were 1 mm stage and monocyte microspore when they were 2 mm stage. Electron microscopy showed that there was significant difference between MS lines and MF (male fertility) lines. In order to detect the global expression of the genes during the melon anther development and association with MS, 12 DEGs (differentially expressed genes) libraries were constructed from the anther of MS and MF in the bud stage with 1 and 2 mm diameter, respectively. A total of 765 DEGs expressed in anther during different developmental stage (MS 1 mm vs. MS 2 mm), 148 and 309 DEGs were found to be related to MS as compared to MF (MS 1 mm vs. MF 1 mm, and MS 2 mm vs. MF 2 mm) at a false discovery rate FDR <0.01. Among these, 10 DEGs were expressed in all the three comparisons, including transcription factor bHLH genes. Among the DEGs in RNA-seq analysis, 28 were validated by qRT-PCR. Of these, a number of genes were involved in ABC transfactor B family, cytochrome-related genes, hormone-related genes (auxin transporter, gibberellin-regulated protein), MADS-box protein genes, F-box protein genes, peroxidase-related, and Zinc finger protein genes. These genes are involved in many biological pathways, including starch and sucrose metabolism, signal transduction mechanisms and transcription factors, etc. Compared to the same developmental stage of MS and MF, the different developmental stages of MS indicated diverse gene regulation pathways involved in the anther development in MS. These results would provide novel insight into the global network to male sterility in melon.

Fine mapping of qTGW10-20.8, a QTL having important contribution to grain weight variation in rice

Grain weight is one of the most important determinants of grain yield in rice. In this study, QTL analysis for grain weight, grain length, and grain width was performed using populations derived from crosses between major parental lines of three-line indica hybrid rice. A total of 27 QTL for grain weight were detected using three recombinant inbred line populations derived from the crosses Teqing/IRBB lines, Zhenshan 97/Milyang 46, and Xieqingzao/Milyang 46. Of these, 10 were found in only a single population and the other 17 in two or all three populations. Nine of the 17 common QTL were located in regions where no QTL associated with grain weight have been cloned and one was selected for fine-mapping. Eight populations segregating in an isogenic background were derived from one F7 residual heterozygote of Teqing/IRBB52. The target QTL, qTGW10-20.8 controlling grain weight, grain length, and grain width, was localized to a 70.7-kb region flanked by InDel markers Te20811 and Te20882 on the long arm of chromosome 10. The QTL region contains seven annotated genes, of which six encode proteins with known functional domains and one encodes a hypothetical protein. One of the genes, Os10g0536100 encoding the MIKC-type MADS-box protein OsMADS56, is the most likely candidate for qTGW10-20.8. These results provide a basis for cloning qTGW10-20.8, which has an important contribution to grain weight variation in rice.

Floral regulators FLC and SOC1 directly regulate expression of the B3-type transcription factor TARGET OF FLC AND SVP 1 at the Arabidopsis shoot apex via antagonistic chromatin modifications.

Integration of environmental and endogenous cues at plant shoot meristems determines the timing of flowering and reproductive development. The MADS box transcription factor FLOWERING LOCUS C (FLC) of Arabidopsis thaliana is an important repressor of floral transition, which blocks flowering until plants are exposed to winter cold. However, the target genes of FLC have not been thoroughly described, and our understanding of the mechanisms by which FLC represses transcription of these targets and how this repression is overcome during floral transition is still fragmentary. Here, we identify and characterize TARGET OF FLC AND SVP1 (TFS1), a novel target gene of FLC and its interacting protein SHORT VEGETATIVE PHASE (SVP). TFS1 encodes a B3-type transcription factor, and we show that tfs1 mutants are later flowering than wild-type, particularly under short days. FLC and SVP repress TFS1 transcription leading to deposition of trimethylation of Iysine 27 of histone 3 (H3K27me3) by the Polycomb Repressive Complex 2 at the TFS1 locus. During floral transition, after downregulation of FLC by cold, TFS1 transcription is promoted by SUPPRESSOR OF OVEREXPRESSION OF CONSTANS1 (SOC1), a MADS box protein encoded by another target of FLC/SVP. SOC1 opposes PRC function at TFS1 through recruitment of the histone demethylase RELATIVE OF EARLY FLOWERING 6 (REF6) and the SWI/SNF chromatin remodeler ATPase BRAHMA (BRM). This recruitment of BRM is also strictly required for SQUAMOSA PROMOTER BINDING PROTEIN-LIKE 9 (SPL9) binding at TFS1 to coordinate RNAPII recruitment through the Mediator complex. Thus, we show that antagonistic chromatin modifications mediated by different MADS box transcription factor complexes play a crucial role in defining the temporal and spatial patterns of transcription of genes within a network of interactions downstream of FLC/SVP during floral transition.

Overexpression of anAPETALA1-like gene from cucumber (Cucumis sativusL.) induces earlier flowering and abnormal leaf development in transgenicArabidopsis

MADS-box proteins are important transcription factors that play essential roles in various aspects of plant development, particularly in flower development. In this study, we performed the identification and functional characterization of CsMADS09 isolated from cucumber (Cucumis sativus L.). CsMADS09 contains a 648-bp open reading frame encoding 215 amino acid residues, and shares high sequence identities with the members of the AP1/FUL family of MADS-box proteins, especially the euAPETALA1 (euAP1) subclade. Many cis-elements related to plant development, stress response, and hormones were identified in the promoter region of CsMADS09. Quantitative real-time polymerase chain reaction results showed that CsMADS09 was mainly expressed in reproductive tissues such as male flowers and unexpanded ovaries, while its expression was low in roots and only traceable in fertilized ovaries. Moreover, the results revealed that CsMADS09 expression tended to decline during male flower development and stayed nearly constant during female flower development. Ectopic expression of CsMADS09 resulted in earlier flowering and abnormal leaf development in transgenic Arabidopsis. This study is the first functional analysis of an AP1-like gene from cucumber and provides some clues for revealing the molecular mechanisms of flower development in cucumber.

The MADS-box genes expressed in the inflorescence of Orchis italica (Orchidaceae).

The Orchidaceae family, which is one of the most species-rich flowering plant families, includes species with highly diversified and specialized flower shapes. The aim of this study was to analyze the MADS-box genes expressed in the inflorescence of Orchis italica, a wild Mediterranean orchid species. MADS-box proteins are transcription factors involved in various plant biological processes, including flower development. In the floral tissues of O. italica, 29 MADS-box genes are expressed that are classified as both class I and II. Class I MADS-box genes include one Mβ-type gene, thereby confirming the presence of this type of MADS-box genes in orchids. The class II MIKC* gene is highly expressed in the column, which is consistent with the conserved function of the MIKC* genes in gametophyte development. In addition, homologs of the SOC, SVP, ANR1, AGL12 and OsMADS32 genes are expressed. Compared with previous knowledge on class II MIKCC genes of O. italica involved in the ABCDE model of flower development, the number of class B and D genes has been confirmed. In addition, 4 class A (AP1/FUL) transcripts, 2 class E (SEP) transcripts, 2 new class C (AG) transcripts and 1 new AGL6 transcript have been identified. Within the AP1/FUL genes, the sequence divergence, relaxation of purifying selection and expression profiles suggest a possible functional diversification within these orchid genes. The detection of only two SEP transcripts in O. italica, in contrast with the 4 genes found in other orchids, suggests that only two SEP genes could be present in the subfamily Orchidoideae. The expression pattern of the MIKCC genes of O. italica indicates that low levels at the boundary of the domain of a given MADS-box gene can overlap with the expression of genes belonging to a different functional A-E class in the adjacent domain, thereby following a “fading borders” model.

Overexpression of both flowering time genes AtSOC1 and SaFUL revealed huge influence onto plant habitus in poplar

The MADS box genes SUPPRESSOR OF OVEREXPRESSION OF CONSTANS1 (SOC1) and FRUITFULL (FUL) are widely known to be involved in the determination of flowering time and the transition from vegetative to reproductive meristem identity in A. thaliana and many other plant species. Analog to the induction of SOC1 and FUL, double mutants in Arabidopsis revealed both secondary cell wall and wood formation. We tested a putative influence of the overexpression of both genes onto growth and plant habitus in the perennial genus Populus. Two different poplar hybrid clones were transformed with SOC1 from A. thaliana (AtSOC1) and FUL from Sinapis alba (SaFUL). Both genes were under the control of the CaMV-35S promoter to induce constitutive overexpression. Regenerated single and double transgenic lines were transferred to the glasshouse and cultivated for up to two vegetation periods. Parallel overexpression of both genes led to plants with a significantly restricted plant growth compared to both untransformed controls and single transgenic plants. Extreme phenotypes also exhibit a different overall habitus of plants and leaf morphology. Thus, both genes seem to be involved in plant habitus and biomass formation in poplar. This study confirms the hypothesis that the two MADS box genes SOC1 and FUL are not only part of reproductive meristem regulation but also involved in vegetative meristem growth. In the next step, downregulation of all SOC1 and FUL genes in poplar by genome editing could lead to increased plant growth and biomass formation.

Floral MADS-box protein interactions in the early diverging angiosperm Aristolochia fimbriata Cham. (Aristolochiaceae: Piperales).

Floral identity MADS-box A, B, C, D, E, and AGL6 class genes are predominantly single copy in Magnoliids, and predate the whole genome duplication (WGD) events in monocots and eudicots. By comparison with the model species Arabidopsis thaliana, the expression patterns of B-, C-, and D-class genes in stamen, carpel, and ovules are conserved in Aristolochia fimbriata, whereas A-, E-class, and AGL6 genes have different expression patterns. Nevertheless, the interactions of these proteins that act through multimeric complexes remain poorly known in early divergent angiosperms. This study evaluates protein interactions among all floral MADS-box A. fimbriata proteins using the Yeast Two Hybrid System (Y2H). We found no homodimers and less heterodimers formed by AfimFUL when compared to AfimAGL6, which allowed us to suggest AGL6 homodimers in combination with AfimSEP2 as the most likely tetramer in sepal identity. We found AfimAP3-AfimPI obligate heterodimers and AfimAG-AfimSEP2 protein interactions intact suggesting conserved stamen and carpel tetrameric complexes in A. fimbriata. We observed a broader interaction partner set for AfimSEP2 than for its paralog AfimSEP1. We show conserved and exclusive MADS-box protein interactions in A. fimbriata in comparison with other eudicot and monocot model species in order to establish plesiomorphic MADS-box protein floral networks in angiosperms.

Identification and characterization of drought responsive miRNAs in a drought tolerant upland rice cultivar KMJ 1-12-3

Shortfall of rain that creates drought like situation in non-irrigated agriculture system often limits rice production, necessitating introduction of drought tolerance trait into the cultivar of interest. The mechanism governing drought tolerance is, however, largely unknown, particularly the involvement of miRNAs, the master regulators of biochemical events. In this regard, response study on a drought tolerant rice variety KMJ 1-12-3 to 20% PEG (osmolality- 315 mOsm/kg) as drought stress revealed significant changes in abundance of several conserved miRNAs targeting transcription factors like homeodomain-leucine zipper, MADS box family protein, C2H2 zinc finger protein and Myb, well known for their importance in drought tolerance in plants. The response study also revealed significant PEG-induced decrease in abundance of the miRNAs targeting cyclin A, cyclin-dependent kinase, guanine nucleotide exchange factor, GTPase-activating protein, 1-aminocyclopropane-1-carboxylic acid oxidase and indole-3-acetic beta-glucosyl transferase indicating miRNA-regulated role of the cell cycle regulators, G-protein signalling and the plant hormones ethylene and IAA in drought tolerance in plants. The study confirmed the existence of four novel miRNAs, including osa-miR12470, osa-miR12471, osa-miR12472 and osa-miR12473, and the targets of three of them could be successfully validated. The PEG-induced decrease in abundance of the novel miRNAs osa-miR12470 and osa-miR12473 targeting RNA dependent RNA polymerase and equilibrative nucleoside transporter, respectively suggested an overall increase in both degradation and synthesis of nucleic acids in plants challenged with drought stress. The drought-responsive miRNAs identified in the study may be proved useful in introducing the trait in the rice cultivars of choice by manipulation of their cellular abundance.

Transcriptomic analysis of Citrus clementina mandarin fruits maturation reveals a MADS-box transcription factor that might be involved in the regulation of earliness

BackgroundHarvest time is a relevant economic trait in citrus, and selection of cultivars with different fruit maturity periods has a remarkable impact in the market share. Generation of early- and late-maturing cultivars is an important target for citrus breeders, therefore, generation of knowledge regarding the genetic mechanisms controlling the ripening process and causing the early and late phenotypes is crucial. In this work we analyze the evolution of the transcriptome during fruit ripening in 3 sport mutations derived from the Fina clementine (Citrus clementina) mandarin: Clemenules (CLE), Arrufatina (ARR) and Hernandina (HER) that differ in their harvesting periods. CLE is considered a mid-season cultivar while ARR and HER are early- and late-ripening mutants, respectively.ResultsWe used RNA-Seq technology to carry out a time course analysis of the transcriptome of the 3 mutations along the ripening period. The results indicated that in these mutants, earliness and lateness during fruit ripening correlated with the advancement or delay in the expression of a set of genes that may be implicated in the maturation process. A detailed analysis of the transcription factors known to be involved in the regulation of fruit ripening identified a member of the MADS box family whose expression was lower in ARR, the early-ripening mutant, and higher in HER, the late-ripening mutant. The pattern of expression of this gene during the maturation period was basically contrary to those of the ethylene biosynthetic genes, SAM and ACC synthases and ACC oxidase. The gene was present in hemizygous dose in the early-ripening mutant.ConclusionsOur analysis provides new clues about the genetic control of fruit ripening in citrus and allowed the identification of a transcription factor that could be involved in the early phenotype.

Isolation and characterization of a MADS-box gene in cucumber (Cucumis sativus L.) that affects flowering time and leaf morphology in transgenic Arabidopsis

The MADS-box family genes are important transcription factors that play essential roles in plant growth and development. In this study, a MADS-box gene, CsMADS02, was identified and functionally characterized in cucumber (Cucumis sativus L.). CsMADS02 encoded a typical SEP MADS-box protein which contained a MADS-box, a K-box, an I region and a C region, as well as two short conserved SEP motifs. Sequence alignment and phylogenetic analysis showed that the CsMADS02 protein belonged to the SEP3 subfamily and was highly homologous to other published plant SEP proteins. Expression analyses revealed that CsMADS02 was primarily expressed in flowers and ovaries, and differentially expressed during the development of male and female flowers. Ectopic overexpression of CsMADS02 caused earlier flowering of Arabidopsis. In addition, morphological changes were also observed in transgenic plants, including curled and fewer rosette leaves. These findings suggest that CsMADS02 is a potential regulator of the flowering time and leaf morphology in cucumber.

Isolation and Functional Characterization of an AGAMOUS-LIKE 18 (AGL18) MADS-box Gene from Cucumber (Cucumis sativus L.)

MADS-box proteins play vital roles in plant growth and development. However, few studies have addressed the biological functions of MADS-box genes in cucumber. In this study, a MADS-box gene, CsMADS25, was cloned from cucumber (Cucumis sativus L.). The open reading frame (ORF) of CsMADS25 was 810 bp in length and encoded a deduced protein consisting of 269 amino acids with a calculated MW of 30.53 kDa and a theoretical pI of 5.38. Sequence alignment showed that CsMADS25 shared the highest amino acid identity with CmMADS09 from Cucumis melo. Phylogenetic tree analysis indicated that CsMADS25 was clustered with AGL18 proteins with high bootstrap values. qRT-PCR analysis showed that the expression of CsMADS25 was observably regulated by various abiotic stresses and GAtreatments. Overexpression of CsMADS25 resulted in dwarf and small-leaf phenotypes in transgenic Arabidopsis plants, and the leaf index value (leaf length/width ratio) of transgenic plants was dramatically increased compared with that of wild-type (WT) plants. These findings suggest that CsMADS25 might play important roles in various developmental processes and in response to abiotic stress of cucumber.

An AGAMOUS MADS-box protein, SlMBP3, regulates the speed of placenta liquefaction and controls seed formation in tomato.

AGAMOUS (AG) MADS-box transcription factors have been shown to play crucial roles in floral organ and fruit development in angiosperms. Here, we isolated a tomato (Solanum lycopersicum) AG MADS-box gene SlMBP3 and found that it is preferentially expressed in flowers and during early fruit developmental stages in the wild-type (WT), and in the Nr (never ripe) and rin (ripening inhibitor) mutants. Its transcripts are notably accumulated in the pistils; transcripts abundance decrease during seed and placental development, increasing again during flower development. SlMBP3-RNAi tomato plants displayed fleshy placenta without locular gel and extremely malformed seeds with no seed coat, while SlMBP3-overexpressing plants exhibited advanced liquefaction of the placenta and larger seeds. Enzymatic activities related to cell wall modification, and the contents of cell wall components and pigments were dramatically altered in the placentas of SlMBP3-RNAi compared with the WT. Alterations in these physiological features were also observed in the placentas of SlMBP3-overexpressing plants. The lignin content of mature seeds in SlMBP3-RNAi lines was markedly lower than that in the WT. RNA-seq and qRT-PCR analyses revealed that genes involved in seed development and the biosynthesis of enzymes related to cell wall modification, namely gibberellin, indole-3-acetic acid, and abscisic acid were down-regulated in the SlMBP3-RNAi lines. Taking together, our results demonstrate that SlMBP3 is involved in the regulation of placenta and seed development in tomato.

A Cucumber AGAMOUS-LIKE 15 (AGL15) MADS-Box Gene Mediates Abnormal Leaf Morphology in Arabidopsis

The AGL15 subfamily MADS-box proteins play vital roles in various developmental processes, such as floral transition, somatic embryogenesis, and leaf and fruit development. In this work, an AtAGL15 ortholog, CsMADS26, was cloned from cucumber (Cucumis sativus L.). The open reading frame (ORF) of CsMADS26 is 669 bp in length, encoding a predicted protein of 222 amino acids. The CsMADS26 protein contains a highly conserved MADS-box domain and a variable C domain, as well as less conserved I and K domains. Phylogenetic relationship analysis revealed that CsMADS26 was clustered into the AGL15 clade of AGL15 subfamily. Expression analysis based on qRT-PCR showed that CsMADS26 is mainly expressed in reproductive organs including flowers and fruits. Transgenic Arabidopsis plants with ectopic expression of CsMADS26 exhibited curled rosette and cauline leaves, and the leaf size was much smaller than that of wild-type (WT) plants. These results provide clues for the functional characterization of CsMADS26 in the future.

A critical evaluation of the role of ethylene and MADS transcription factors in the network controlling fleshy fruit ripening.

Contents Summary 1724 I. Introduction 1725 II. Ripening genes 1725 III. The importance of ethylene in controlling ripening 1727 IV. The importance of MADS-RIN in controlling ripening 1729 V. Interactions between components of the ripening regulatory network 1734 VI. Conclusions 1736 Acknowledgements 1738 Author contributions 1738 References 1738 SUMMARY: Understanding the regulation of fleshy fruit ripening is biologically important and provides insights and opportunities for controlling fruit quality, enhancing nutritional value for animals and humans, and improving storage and waste reduction. The ripening regulatory network involves master and downstream transcription factors (TFs) and hormones. Tomato is a model for ripening regulation, which requires ethylene and master TFs including NAC-NOR and the MADS-box protein MADS-RIN. Recent functional characterization showed that the classical RIN-MC gene fusion, previously believed to be a loss-of-function mutation, is an active TF with repressor activity. This, and other evidence, has highlighted the possibility that MADS-RIN itself is not important for ripening initiation but is required for full ripening. In this review, we discuss the diversity of components in the control network, their targets, and how they interact to control initiation and progression of ripening. Both hormones and individual TFs affect the status and activity of other network participants, which changes overall network signaling and ripening outcomes. MADS-RIN, NAC-NOR and ethylene play critical roles but there are still unanswered questions about these and other TFs. Further attention should be paid to relationships between ethylene, MADS-RIN and NACs in ripening control.

Genome-wide identification of long non-coding RNA targets of the tomato MADS box transcription factor RIN and function analysis.

In recent years, increasing numbers of long non-coding RNAs (lncRNAs) have been identified in humans, animals and plants, and several of them have been shown to play important roles in diverse biological processes. However, little work has been performed on the regulation mechanism of lncRNA biogenesis and expression, especially in plants. Compared with studies of tomato MADS-box transcription factor RIPENING INHIBITOR (RIN) target coding genes, there are few reports on its relationship to non-coding RNAs. The aim of the present study was to identify and explore the specific role of RIN target lncRNAs in tomato fruit development and ripening. lncRNA targets of RIN were identified by chromatin immunoprecipitation sequencing (ChIP-seq) combined with RNA deep sequencing analysis. Six selected lncRNA targets were validated by quantitative real-time PCR, ChIP and electrophoretic mobility shift assays, and we further confirmed differential expression between wild-type and ripening-deficient mutant fruit, and RIN direct binding in the promoter regions. By means of virus-induced gene silencing (VIGS) assays and a clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) genome editing strategy, the ripening-related function of a specific target lncRNA (lncRNA2155) was studied. We identified 187 lncRNAs as direct RIN targets, which exhibited RIN binding sites in their promoters and showed different expression between the wild-type and rin mutant. Six target lncRNAs were shown to bind with RIN directly in their promoters in vivo and in vitro. Moreover, using CRISPR/Cas9 technology to knock out the locus of the target lncRNA2155 indicated that it delayed fruit ripening in tomato. Collectively, these findings provide new insight into RIN in the transcriptional regulation of lncRNAs and suggest that lncRNAs will contribute to a better understanding of the RIN regulatory network that controls fruit ripening.

A MADS-box transcription factor regulates a central step in sporulation of the oomycete Phytophthora infestans.

MADS-box transcription factors play significant roles in eukaryotes, but have not yet been characterized in oomycetes. Here, we describe a MADS-box protein from Phytophthora infestans, which causes late blight of potato. P. infestans and most other oomycetes express a single MADS-box gene. PiMADS is not transcribed during vegetative growth, but is induced early during asexual sporulation. Its mRNA levels oscillate in response to light, which suppresses sporulation. The protein was not detected in nonsporulating mycelia, but was found in sporulating mycelia and spores. Both mRNA and protein levels decline upon spore germination. A similar expression pattern as well as nuclear localization was observed when the protein was expressed with a fluorescent tag from the native promoter. Gene silencing triggered by a construct expressing 478 nt of MADS sequences indicated that PiMADS is required for sporulation but not hyphal growth or plant colonization. A comparison of wild type to a silenced strain by RNA-seq indicated that PiMADS regulates about 3000 sporulation-associated genes, and acts before other genes previously shown to regulate sporulation. Analysis of the silenced strain also indicated that the native gene was not transcribed while the transgene was still expressed, which contradicts current models for homology-dependent silencing in oomycetes.

From microinjection to genome editing using zinc finger nucleases and CRISPR/Cas9 system. Characteristics of transgenic pigs generated for xenotransplantation purposes

Identification of genes expressed during the Golovinomyces orontii infection process in Helianthus niveus assumes importance for incorporation of resistance to powdery mildew in cultivated sunflower (H. annuus L.) from this donor species. RNA-seq analysis of control (uninfected) and infected samples of H. niveus resulted in identification of 231,754 transcripts. A total of 3726 transcripts were differentially expressed of which 205 were specifically expressed in control and 1961 in infected samples. Functional annotation of the differentially expressed transcripts showed significant upregulation of GRAS type transcription factor (TF) and plant specific GATA-type zinc finger TF in infected samples and the K-box, MADS box TF and WRKY family TF in control tissues. Gene ontology classification showed that genes involved in cell and cell part functioning, catalytic and metabolic processes were significantly and highly expressed. This is the first application of RNA-Seq for identification of key genes and pathways involved in powdery mildew infection process in a Helianthus species conferring resistance to the pathogen.

Genome-Wide Association Analysis of Mucilage and Hull Content in Flax (Linum usitatissimum L.) Seeds.

New flaxseed cultivars differing in seed mucilage content (MC) with low hull content (HC) represent an attractive option to simultaneously target the food and feed markets. Here, a genome-wide association study (GWAS) was conducted for MC and HC in 200 diverse flaxseed accessions genotyped with 1.7 million single nucleotide polymorphism (SNP) markers. The data obtained for MC and HC indicated a broad phenotypic variation and high (~70%) and a moderate (~49%) narrow sense heritability, respectively. MC and HC did not differ statistically between fiber and oil morphotypes, but yellow-seeded accessions had 2.7% less HC than brown-seeded ones. The genome-wide linkage disequilibrium (LD) decayed to r2 = 0.1 at a physical distance of ~100 kb. Seven and four quantitative trait loci (QTL) were identified for MC and HC, respectively. Promising candidate genes identified include Linum usitatissimum orthologs of the Arabidopsis thaliana genes TRANSPARENT TESTA 8, SUBTILISIN-LIKE SERINE PROTEASE, GALACTUROSYL TRANSFERASE-LIKE 5, MUCILAGE-MODIFIED 4, AGAMOUS-LIKE MADS-BOX PROTEIN AGL62, GLYCOSYL HYDROLASE FAMILY 17, and UDP-GLUCOSE FLAVONOL 3-O-GLUCOSYLTRANSFERASE. These genes have been shown to play a role in mucilage synthesis and release, seed coat development and anthocyanin biosynthesis in A. thaliana. The favorable alleles will be useful in flaxseed breeding towards the goal of achieving the ideal MC and HC composition for food and feed by genomic-based breeding.

The Floral C-Lineage Genes Trigger Nectary Development in Petunia and Arabidopsis

To attract insects, flowers produce nectar, an energy-rich substance secreted by specialized organs called nectaries. For Arabidopsis thaliana, a rosid species with stamen-associated nectaries, the floral B-, C-, and E-functions were proposed to redundantly regulate nectary development. Here, we investigated the molecular basis of carpel-associated nectary development in the asterid species petunia (Petunia hybrida). We show that its euAGAMOUS (euAG) and PLENA (PLE) C-lineage MADS box proteins are essential for nectary development, while their overexpression is sufficient to induce ectopic nectaries on sepals. Furthermore, we demonstrate that Arabidopsis nectary development also fully depends on euAG/PLE C-lineage genes. In turn, we show that petunia nectary development depends on two homologs of CRABS CLAW (CRC), a gene previously shown to be required for Arabidopsis nectary development, and demonstrate that CRC expression in both species depends on the members of both euAG/PLE C-sublineages. Therefore, petunia and Arabidopsis employ a similar molecular mechanism underlying nectary development, despite otherwise major differences in the evolutionary trajectory of their C-lineage genes, their distant phylogeny, and different nectary positioning. However, unlike in Arabidopsis, petunia nectary development is position independent within the flower. Finally, we show that the TARGET OF EAT-type BLIND ENHANCER and APETALA2-type REPRESSOR OF B-FUNCTION genes act as major regulators of nectary size.

Analysis of long-non-coding RNAs associated with ethylene in tomato

Long-Non-Coding RNAs (LncRNAs) are a class of non-coding endogenous RNAs contributing to numerous biological processes. LeERF1 is a tomato ethylene response factor (ERF) near the end of the ethylene signal transduction pathway. To identify lncRNAs in tomato and elucidate their roles in ethylene signaling, deep sequencing was deployed in over-expression and repression LeERF1 transgenic and control tomato fruits. A total of 397 lncRNAs were identified, including 169 tomato lncRNAs that had not previously been identified. Among these, 12 were differentially expressed between the transgenic and control tomato fruits. Numerous lncRNA target genes were identified including many associated with ethylene signaling including auxin response factors and auxin-induced proteins, F-box proteins, ERFs and MADS-box proteins. In addition, two lncRNAs were found to be the precursor of three miRNAs and four lncRNAs could be targeted by five miRNAs. We propose a regulatory model highlighting the relationships between lncRNAs and their targets involved in ethylene signal transduction which establishes a foundation for addressing the role of LncRNAs in ethylene response.