Development of 50 InDel-based barcode system for genetic identification of tartary buckwheat resources.

Tartary buckwheat (Fagopyrum tataricum Gartn.) is a highly functional crop that is poised to be the target of many future breeding efforts. The reliable ex situ conservation of various genetic resources is essential for the modern breeding of tartary buckwheat varieties. We developed PCR-based co-dominant insertion/deletion (InDel) markers to discriminate tartary buckwheat genetic resources. First, we obtained the whole genome from 26 accessions across a superscaffold-scale reference genome of 569.37 Mb for tartary buckwheat cv. "Daegwan 3-7." Next, 171,926 homogeneous and 53,755 heterogeneous InDels were detected by comparing 26 accessions with the "Daegwan 3-7" reference sequence. Of these, 100 candidate InDels ranging from 5-20 bp in length were chosen for validation, and 50 of them revealed polymorphisms between the 26 accessions and "Daegwan 3-7." The validated InDels were further tested through the assessment of their likelihood to give rise to a single or a few PCR products in 50 other accessions, covering most tartary buckwheat genome types. The major allele frequencies ranged from 0.5616 at the TB42 locus to 0.9863 at the TB48 locus, with the average PIC value of 0.1532 with a range of 0.0267-0.3712. To create a user-friendly system, the homology of the genotypes between and among the accessions were visualized in both one- (1D) and two-dimensional (2D) barcode types by comparing amplicon polymorphisms with the reference variety, "Daegwan 3-7." A phylogenetic tree and population structure of the 76 accessions according to amplicon polymorphisms for the 50 InDel markers corresponded to those using non-synonymous single nucleotide polymorphism variants, indicating that the barcode system based on the 50 InDels was a useful tool to improve the reliability of identification of tartary buckwheat accessions in the germplasm stocks.

Cytochrome P450 family: Genome-wide identification provides insights into the rutin synthesis pathway in Tartary buckwheat and the improvement of agricultural product quality.

Flavonoids can not only help plants resist ultraviolet and pathogen attacks, but also show a wide range of therapeutic prospects for human health, including antioxidant, anti-inflammatory and anti-hypertension. Tartary buckwheat, as medicinal and food homologous crop, is rich in flavonoids, among which rutin may prevent liver damage. The one of the major objectives of Tartary buckwheat breeding is to cultivate varieties that have large fruits, high flavonoids and nutrient contents. Members of the cytochrome P450 monooxygenase (CYP) superfamily play a vital role in the synthesis of flavonoids, plant growth and development. Whole-genome analyses of the CYP family have been performed in several plants, but the CYP family has not been characterized in Tartary buckwheat. In this study, 285 FtCYPs were identified from the genome to improve the rutin content and quality of Tartary buckwheat. By exploring the structure, motif composition, tandem and segmental duplication events of FtCYPs, as well as evolutionary relationships with CYPs in other plants, we preliminarily screened potential FtCYPs regulating rutin synthesis, growth and development. The expression levels of the FtCYPs in different organs and fruits at various periods were measured. This study provides a solid foundation for verifying the function of FtCYPs, cultivating high rutin Tartary buckwheat varieties.

Proximate composition, amino acid profile, pasting and process characteristics of flour from different Tartary buckwheat varieties.

Flour from twenty-three Tartary buckwheat varieties were evaluated and compared for proximate composition, mineral and amino acid profile. Further, pasting properties and process characteristics such as foaming, oil and water absorption capacities, emulsification properties were determined for identifying the efficient application of Tartary buckwheat in food systems. Ash, protein and fat contents of the flours ranged between 1.76-2.80%, 9.06-14.88%, and 2.02-3.60%, respectively. Buckwheat flours from all varieties had abundant K, Mg and Ca content, with the highest in B-121, IC-329200 and IC-274439, respectively. All essential amino acids were detected in all varieties with leucine present in abundance. Isoleucine, cystine and asparagine were limiting. Emulsifying and foaming properties of all buckwheat flours improved as the pH increased from 4 to 10. Emulsion activity index (pH 10) showed a significant positive correlation with hydrophobic amino acids. Flours from most of the buckwheat varieties had unique pasting properties with very low set back and breakdown viscosities indicating paste stability and lower retrogradation tendency, making them suitable for thickening of sauces and soups.

Genome-wide investigation of the heat shock transcription factor (Hsf) gene family in Tartary buckwheat (Fagopyrum tataricum).

BACKGROUND: Heat shock transcription factor (Hsfs) is widely found in eukaryotes and prokaryotes. Hsfs can not only help organisms resist high temperature, but also participate in the regulation of plant growth and development (such as involved in the regulation of seed maturity and affects the root length of plants). The Hsf gene was first isolated from yeast and then gradually found in plants and sequenced, such as Arabidopsis thaliana, rice, maize. Tartary buckwheat is a rutin-rich crop, and its nutritional value and medicinal value are receiving more and more attention. However, there are few studies on the Hsf genes in Tartary buckwheat. With the whole genome sequence of Tartary buckwheat, we can effectively study the Hsf gene family in Tartary buckwheat. RESULTS: According to the study, 29 Hsf genes of Tartary buckwheat (FtHsf) were identified and renamed according to location of FtHsf genes on chromosome after removing a redundant gene. Therefore, only 29 FtHsf genes truly had the functional characteristics of the FtHsf family. The 29 FtHsf genes were located on 8 chromosomes of Tartary buckwheat, and we found gene duplication events in the FtHsf gene family, which may promote the expansion of the FtHsf gene family. Then, the motif compositions and the evolutionary relationship of FtHsf proteins and the gene structures, cis-acting elements in the promoter, synteny analysis of FtHsf genes were discussed in detail. What's more, we found that the transcription levels of FtHsf in different tissues and fruit development stages were significantly different by quantitative real-time PCR (qRT-PCR), implied that FtHsf may differ in function. CONCLUSIONS: In this study, only 29 Hsf genes were identified in Tartary buckwheat. Meanwhile, we also classified the FtHsf genes, and studied their structure, evolutionary relationship and the expression pattern. This series of studies has certain reference value for the study of the specific functional characteristics of Tartary buckwheat Hsf genes and to improve the yield and quality of Tartary buckwheat in the future.

psbE-psbL and ndhA Intron, the Promising Plastid DNA Barcode of Fagopyrum.

Buckwheat is an important functional food material with high nutritional value. However, it is still a difficult task for the taxonomy studies of wild buckwheat that are only based on morphology. In order to demonstrate the most efficient DNA barcode in the phylogenetic research of buckwheat, promote the investigation of wild buckwheat, and also reveal the phylogenetic relationship between Fagopyrum species, psbE-psbL and ndhA intron were validated here, which previously have been proved to be promising DNA barcode candidates for phylogenetic studies in genera Fagopyrum. Meanwhile, ndhA intron + psbE-psbL and matK + psbE-psbL could distinguish the relationship between species clearly. Combining the results of morphology and molecular markers, we suggested the buckwheat species should be divided into two subgroups, one subgroup consisted of F. tataricum, F. esculentum, F. cymosum and its related wild species, and the other subgroup included other wild buckwheat species. Our results could fulfill molecular markers of taxonomy research in genera Fagopyrum, promote wild buckwheat species identification, and assist in the use of wild buckwheat resources in the future. Additionally, the phylogenetic relationship revealed here could provide valuable information for molecular breeding of buckwheat and provide reference for inter-species hybridization.

[Screening genotypes and identifying indicators of different Fagopyrum tataricum varieties with low phosphorus tolerance.] / è‹¦èžè€ä½Žç£·åŸºå› åž‹ç­›é€‰åŠè¯„ä»·æŒ‡æ ‡çš„é‰´å®š.

Soil phosphorus (P) deficiency is one of the main factors that diminish the yield of crops on the Loess Plateau. Fagopyrum tataricum is a dominant cereal plant on the Loess Plateau, whose responses to low-P stress are significantly different and dependent on its genotypes. Therefore, screening genotypes with efficient P utilization is an important approach to increase the yield of F. tataricum and promote the sustainable development of local agriculture. With fourteen F. tataricum genotypes, the agronomic traits, physiology and biochemical characters of F. tataricum at seedling stage under normal (2 mmol·L-1) and low-P treatment (0.2 mmol·L-1) were studied by using sand culture. By examining the characters of selected F. tataricum at seedling stage, the varieties of F. tataricum with low-P tolerance were screened out and their evaluation indices were assessed to provide theoretical basis for the breeding of F. tataricum with high P-efficiency and the infertile soil cultivation on the Loess Plateau. The results showed that under low-P stress, the shoots of all genotypes were more affected than roots, with the decline of shoot aboveground index, average root diameter, root surface area and root volume, whereas the length of main root was elongated. Root activity and soluble protein content were decreased. In contrast, other physiological and biochemical indicators were increased. Moreover, plant total phosphorus content and plant phosphorus accumulation reduced, but phosphorus use efficiency increased. The principal component analysis divided the 22 individual indicators into four mutually independent comprehensive indicators (cumulative contribution rate of 90.1%). Cluster analysis divided 14 kinds of F. tataricum into three categories: Low phosphorus tolerant, intermediate and phosphorus sensitive. In order to investigate the index of low-P tolerance of F. tataricum at seedling stage, the optimal regression equation was established with the comprehensive evaluation value of low-P tolerance (D value) as the dependent variable and the low-P tolerance index of each indicator as the independent variables. Seven indicators of root surface area, root length, plant height, aboveground dry mass, acid phosphatase, phosphorus accumulation and POD activity were examined, which could be used for the rapid identification of low-P tolerance of F. tataricum at seedling stage.

Determining the geographical origin of common buckwheat from China by multivariate analysis based on mineral elements, amino acids and vitamins.

This study aimed to establish a method for distinguishing the geographical origin of common buckwheat from Inner Mongolia, Shanxi and Shaanxi Provinces in China. Three chemical families including mineral elements, vitamins and amino acids of 48 samples from different geographical origins were analyzed by principal component analysis (PCA), cluster analysis (CA) and linear discriminate analysis (LDA) for this purpose. LDA clearly discriminated the geographical origin of common buckwheat samples grown in three regions, and gave a high correct classification rate of 95.8% and satisfactory cross-validation rate of 91.7%. Some variables (Mn, VPP, Se, Gly, Cu, Asp, Fe, and Ala) significantly contributed to the ability to discriminate the geographical origin of the common buckwheat. These results demonstrated that the proposed method is a powerful tool for controlling the geographical origin of common buckwheat by governmental administration and protecting consumers from improper domestic labeling. However, the discriminant method still needs to be further validated using more reliable data.

Comparative Analysis of Four Buckwheat Species Based on Morphology and Complete Chloroplast Genome Sequences.

Buckwheat is a nutritional and economically crop belonging to Polygonaceae, Fagopyrum. To better understand the mutation patterns and evolution trend in the chloroplast (cp) genome of buckwheat, and found sufficient number of variable regions to explore the phylogenetic relationships of this genus, two complete cp genomes of buckwheat including Fagopyrum dibotrys (F. dibotrys) and Fagopyrum luojishanense (F. luojishanense) were sequenced, and other two Fagopyrum cp genomes were used for comparative analysis. After morphological analysis, the main difference among these buckwheat were height, leaf shape, seeds and flower type. F. luojishanense was distinguishable from the cultivated species easily. Although the F. dibotrys and two cultivated species has some similarity, they different in habit and component contents. The cp genome of F. dibotrys was 159,320 bp while the F. luojishanense was 159,265 bp. 48 and 61 SSRs were found in F. dibotrys and F. luojishanense respectively. Meanwhile, 10 highly variable regions among these buckwheat species were located precisely. The phylogenetic relationships among four Fagopyrum species based on complete cp genomes was showed. The results suggested that F. dibotrys is more closely related to Fagopyrum tataricum. These data provided valuable genetic information for Fagopyrum species identification, taxonomy, phylogenetic study and molecular breeding.

The complete chloroplast genome sequence of Fagopyrum cymosum.

Fagopyrum cymosum is a traditional medicinal plant. In this study, the complete chloroplast genome of Fagopyrum cymosum is presented. The total genome size is 160,546 bp in length, containing a pair of inverted repeats (IRs) of 32,598 bp, separated by large single copy (LSC) and small single copy (SSC) of 84,237 bp and 11,014 bp, respectively. Overall GC contents of the genome were 36.9%. The chloroplast genome harbors 126 annotated genes, including 91 protein coding genes, 29 tRNA genes, and six rRNA genes. Eighteen genes contain one or two introns. Phylogenetic analyses indicated a clear evolutionary relationship among species of Caryophyllales.

Complete Chloroplast Genome Sequence of Tartary Buckwheat (Fagopyrum tataricum) and Comparative Analysis with Common Buckwheat (F. esculentum).

We report the chloroplast (cp) genome sequence of tartary buckwheat (Fagopyrum tataricum) obtained by next-generation sequencing technology and compared this with the previously reported common buckwheat (F. esculentum ssp. ancestrale) cp genome. The cp genome of F. tataricum has a total sequence length of 159,272 bp, which is 327 bp shorter than the common buckwheat cp genome. The cp gene content, order, and orientation are similar to those of common buckwheat, but with some structural variation at tandem and palindromic repeat frequencies and junction areas. A total of seven InDels (around 100 bp) were found within the intergenic sequences and the ycf1 gene. Copy number variation of the 21-bp tandem repeat varied in F. tataricum (four repeats) and F. esculentum (one repeat), and the InDel of the ycf1 gene was 63 bp long. Nucleotide and amino acid have highly conserved coding sequence with about 98% homology and four genes--rpoC2, ycf3, accD, and clpP--have high synonymous (Ks) value. PCR based InDel markers were applied to diverse genetic resources of F. tataricum and F. esculentum, and the amplicon size was identical to that expected in silico. Therefore, these InDel markers are informative biomarkers to practically distinguish raw or processed buckwheat products derived from F. tataricum and F. esculentum.

Comparison of phenolic profiles and antioxidant properties of European Fagopyrum esculentum cultivars.

The purpose of this study was to investigate composition and content of phenolic compounds in seeds of common buckwheat (Fagopyrum esculentum Moench) cultivars from Western, Central and Southeastern Europe grown in the Balkan area, and to compare them with cultivars from the Balkan. Mostly detected hydroxycinnamic acids in seeds of the investigated cultivars were caffeic and chlorogenic acid derivatives. More than ten different flavanols were detected in the investigated seeds, based on which all tested buckwheat cultivars were divided into two groups: those with high propelargonidins (epiafzelechin-epicatechin) and those with high procyanidins contents. 'Novosadska' had the highest level of phenolic acids, proanthocyanidins, flavones and most of the flavonols. However, 'Bosna 1' and 'Bosna 2' were highlighted with the greatest rutin content (up to 46 times higher than in other cultivars). All buckwheat cultivars had quite high antioxidant capacity (more than 80% of neutralized radicals), yet, 'Novosadska', 'Godijevo', 'Spacinska 1' and 'Bamby' excelled.

Comparative analysis of flavonoids and polar metabolite profiling of Tanno-original and Tanno-high rutin buckwheat.

Rutin is an important indicator for evaluating the quality of buckwheat. In this study, flavonoid biosynthesis was compared between two common cultivars (an original and a high-rutin line) of buckwheat, Fagopyrum esculentum Moench. Transcriptional levels of the main flavonoid biosynthetic genes were analyzed by real-time PCR, and main flavonoid metabolites were detected by high-performance liquid chromatography (HPLC); levels of gene expression varied among organs of the two cultivars. Significantly higher transcription levels of most flavonoid biosynthetic genes, except FeFLS1, were detected in stems of the high-rutin line than in stems of the original line. FeCHI and FeFLS2 genes also showed higher expression levels in seeds of the high-rutin cultivar. In contrast, FePAL, FeC4H, Fe4CL1, FeCHS, FeF3H, FeF3'H, FeFLS2, and FeDFR were highly detected in the roots of the original line. The HPLC results indicated 1.73-, 1.62-, and 1.77-fold higher accumulation of rutin (the primary flavonoid compound) in leaves, stems, and mature seeds of the high-rutin cultivar (24.86, 1.46, and 1.36 µg/mg, respectively) compared with the original cultivar (14.40, 0.90, and 0.77 µg/mg, respectively). A total of 46 metabolites were identified from seeds by gas chromatography-time-of-flight mass spectrometry. The metabolite profiles were subjected to principal component analysis (PCA). PCA could clearly differentiate the original and high-rutin cultivars. Our results indicate that the high-rutin cultivar could be an excellent alternative for buckwheat culture, and we provide useful information for obtaining this cultivar.

Metabolomic analysis and differential expression of anthocyanin biosynthetic genes in white- and red-flowered buckwheat cultivars (Fagopyrum esculentum).

Red-flowered buckwheat ( Fagopyrum esculentum ) is used in the production of tea, juice, and alcohols after the detoxification of fagopyrin. In order to investigate the metabolomics and regulatory of anthocyanin production in red-flowered (Gan-Chao) and white-flowered (Tanno) buckwheat cultivars, quantitative real-time RT-PCR (qRT-PCR), gas chromatography time-of-flight mass spectrometry (GC-TOFMS), and high performance liquid chromatography (HPLC) were conducted. The transcriptions of FePAL, FeC4H, Fe4CL1, FeF3H, FeANS, and FeDFR increased gradually from flowering stage 1 and reached their highest peaks at flowering stage 3 in Gan-Chao flower. In total 44 metabolites, 18 amino acids, 15 organic acids, 7 sugars, 3 sugar alcohols, and 1 amine were detected in Gan-Chao flowers. Two anthocyanins, cyanidin 3-O-glucoside and cyanidin 3-O-rutinoside, were identified in Gan-Chao cultivar. The first component of the partial least-squares to latent structures-discriminate analysis (PLS-DA) indicated that high amounts of phenolic, shikimic, and pyruvic acids were present in Gan-Chao. We suggest that transcriptions of genes involved in anthocyanin biosynthesis, anthocyanin contents, and metabolites have correlation in the red-flowered buckwheat Gan-Chao flowers. Our results may be helpful to understand anthocyanin biosynthesis in red-flowered buckwheat.

On the origin of the woody buckwheat Fagopyrum tibeticum (=Parapteropyrum tibeticum) in the Qinghai-Tibetan Plateau.

Here we tested whether 'insular woodiness', a striking evolutionary pattern that commonly occurs on islands, has also appeared in QTP continental endemics. Parapteropyrum, a monotypic shrubby genus occurring in the central QTP, has been previously placed in the tribe Atraphaxideae of the family Polygonaceae, while all the other woody species of this tribe mainly occur in western and central Asia. We studied sequence variations of nuclear ITS (internal transcribed spacer) and cp (chloroplast) DNA (rbcL and accD) of this genus and the other ten genera. The constructed phylogenies based on ITS, cpDNA or a combination of both datasets, suggest that the woody Parapteropyrum is nested within and most likely evolved from the herbaceous Fagopyrum. We propose that the large-scale uplift of the QTP not only promoted continental species radiation, but also the secondary feature of woodiness in a few herbaceous lineages in response to strong selection pressures, similar to those acting on island flora. In addition, the confirmation of Parapteropyrum within Fagopyrum highlights its potential use as a new, perennial source of buckwheat.

De novo sequencing and characterization of floral transcriptome in two species of buckwheat (Fagopyrum).

BACKGROUND: Transcriptome sequencing data has become an integral component of modern genetics, genomics and evolutionary biology. However, despite advances in the technologies of DNA sequencing, such data are lacking for many groups of living organisms, in particular, many plant taxa. We present here the results of transcriptome sequencing for two closely related plant species. These species, Fagopyrum esculentum and F. tataricum, belong to the order Caryophyllales--a large group of flowering plants with uncertain evolutionary relationships. F. esculentum (common buckwheat) is also an important food crop. Despite these practical and evolutionary considerations Fagopyrum species have not been the subject of large-scale sequencing projects. RESULTS: Normalized cDNA corresponding to genes expressed in flowers and inflorescences of F. esculentum and F. tataricum was sequenced using the 454 pyrosequencing technology. This resulted in 267 (for F. esculentum) and 229 (F. tataricum) thousands of reads with average length of 341-349 nucleotides. De novo assembly of the reads produced about 25 thousands of contigs for each species, with 7.5-8.2× coverage. Comparative analysis of two transcriptomes demonstrated their overall similarity but also revealed genes that are presumably differentially expressed. Among them are retrotransposon genes and genes involved in sugar biosynthesis and metabolism. Thirteen single-copy genes were used for phylogenetic analysis; the resulting trees are largely consistent with those inferred from multigenic plastid datasets. The sister relationships of the Caryophyllales and asterids now gained high support from nuclear gene sequences. CONCLUSIONS: 454 transcriptome sequencing and de novo assembly was performed for two congeneric flowering plant species, F. esculentum and F. tataricum. As a result, a large set of cDNA sequences that represent orthologs of known plant genes as well as potential new genes was generated.

[Molecular cloning and prokaryotic expression of phenylalanine ammonia-lyase gene FdPAL from Fagopyrum dibotrys].

OBJECTIVE: To clone and characterize the DNA and cDNA sequences of phenylalanine ammonia-lyase gene (PAL) from Fagopyrum dibotrys, and investigate the biological activity of the obtained PAL. METHOD: Using homology cloning and RT-PCR techniques, the DNA and full-length cDNA sequences of PAL gene were amplified from F. dibotrys. The obtained sequences were analyzed by bioinformatics software. The ORF of PAL gene was cloned into expression vector pET-30b(+) and transformed into Escherichia coli BL21 (DE3) for expression the recombined protein. The catalytic activity of the recombined protein was determined by Spectrophotometer and thin layer chromatography (TLC) methods. RESULT: The DNA sequence of PAL gene (designated as FdPAL, GenBank accession number: HM628904) was 2 583 bp in size, of which consisted two extrons and a single intron, and the full-length cDNA of FdPAL was 2 169 bp in size, which contained an ORF. The deduced protein of FdPAL contained 722 amino acids with calculated molecular weight (MW) of 78.31 kDa and an isoelectric point (pI) of 5.94. The SDS-PAGE results showed that the molecular weight of recombinant FdPAL protein was 75.37 kDa, which is consistent with the predictions. After 4 hours of induction, the enzymatic specific activity of FdPAL reached the summit, up to 4 386 nmol x g(-1) x min(-1). The reaction products were also identified by TLC, using L-Phe and trans-cinnamic acid as the internal standard. CONCLUSION: The PAL gene (both DNA sequence and full-length cDNA sequence) was cloned from F. dibotrys, and it has the same classic characters as other PALs in plants. The recombinant FdPAL was efficiently expressed in E. coli and had the activity for catalyzing the conversion from L-phenylalanine to cinnamic acid.

Genetic mapping of common buckwheat using DNA, protein and morphological markers.

Genetic mapping of F(2) progeny (n = 225) of hybrids between the Sobano variety of common buckwheat (Fagopyrum esculentum Moench) and the Homo wild accession (F. esculentum var. homotropicum) was carried out using randomly amplified polymorphic DNA (RAPD), sequence-tagged site (STS) and seed protein subunit markers, and three morphological traits. Ten linkage groups were identified, involving 87 RAPD markers, 12 STS markers, four seed protein subunit (PS(62)/PS(59), PS(49.8)/PS(51.4), PS(44)/PS(42.9), and PS(39.9)/PS(37.8)) markers, and three morphological alleles controlling homo/long style (H/s), shattering habit (Sht/sht), and acute/obtuse achene ridge (Ac/ac), covering a total of 655.2 cM.

Genetic diversity of storage proteins in cultivated buckwheat.

Prolamin and albumn variations of the storage proteins in 76 cultivated buckwheat accessions (55 accessions of Fagopyrum tataricum, 21 accessions of F. esculentum) from 7 countries were characterized by A-PAGE and SDS-PAGE, respectively, for the purpose of evaluating the genetic diversity of cultivated buckwheat at the level of proteins. A total of 18 prolamin bands were detected, among which 88.89 % bands were polymorphic. The number of albumn bands based on SDS-PAGE observed in accessions ranged from 4 to 10. Most intense bands were in the range of molecular weights from 29 to 97.2 kDa. The average of genetic similarity coefficient based on prolamin bands was 0.784 (in F. tataricum and F. esculentum were 0.892 and 0.681, respectively), while on prolamin and albumn bands was 0.742 (in F. tataricum and F. esculentum were 0.864 and 0.633, respectively). Accessions of F. tataricum and F. esculentum showed significant interspecific variation in the A-PAGE and SDS-PAGE profile of the storage proteins. The cluster analysis indicated that all the accessions could be divided into 3 groups and 3 subgroups. The genetic variations among cultivated buckwheat accessions were associated with their geographic origins in some degree.

Mycorrhizal status and diversity of fungal endophytes in roots of common buckwheat (Fagopyrum esculentum) and tartary buckwheat (F. tataricum).

To determine the mycorrhizal status and to identify the fungi colonising the roots of the plants, common buckwheat (Fagopyrum esculentum) and tartary buckwheat (F. tataricum) were inoculated with an indigenous fungal mixture from a buckwheat field. Root colonisation was characterised by the hyphae and distinct microsclerotia of dark septate endophytes, with occasional arbuscules and vesicles of arbuscular mycorrhizal fungi. Sequences of arbuscular mycorrhizal fungi colonising tartary buckwheat clustered close to the Glomus species group A. Sequences with similarity to the Ceratobasidium/Rhizoctonia complex, a putative dark septate endophyte fungus, were amplified from the roots of both common and tartary buckwheat. To the best of our knowledge, this is the first report of arbuscular mycorrhizal colonisation in tartary buckwheat and the first molecular characterisation of these fungi that can colonise both of these economically important plant species.

Comparative chloroplast genomics and phylogenetics of Fagopyrum esculentum ssp. ancestrale -a wild ancestor of cultivated buckwheat.

BACKGROUND: Chloroplast genome sequences are extremely informative about species-interrelationships owing to its non-meiotic and often uniparental inheritance over generations. The subject of our study, Fagopyrum esculentum, is a member of the family Polygonaceae belonging to the order Caryophyllales. An uncertainty remains regarding the affinity of Caryophyllales and the asterids that could be due to undersampling of the taxa. With that background, having access to the complete chloroplast genome sequence for Fagopyrum becomes quite pertinent. RESULTS: We report the complete chloroplast genome sequence of a wild ancestor of cultivated buckwheat, Fagopyrum esculentum ssp. ancestrale. The sequence was rapidly determined using a previously described approach that utilized a PCR-based method and employed universal primers, designed on the scaffold of multiple sequence alignment of chloroplast genomes. The gene content and order in buckwheat chloroplast genome is similar to Spinacia oleracea. However, some unique structural differences exist: the presence of an intron in the rpl2 gene, a frameshift mutation in the rpl23 gene and extension of the inverted repeat region to include the ycf1 gene. Phylogenetic analysis of 61 protein-coding gene sequences from 44 complete plastid genomes provided strong support for the sister relationships of Caryophyllales (including Polygonaceae) to asterids. Further, our analysis also provided support for Amborella as sister to all other angiosperms, but interestingly, in the bayesian phylogeny inference based on first two codon positions Amborella united with Nymphaeales. CONCLUSION: Comparative genomics analyses revealed that the Fagopyrum chloroplast genome harbors the characteristic gene content and organization as has been described for several other chloroplast genomes. However, it has some unique structural features distinct from previously reported complete chloroplast genome sequences. Phylogenetic analysis of the dataset, including this new sequence from non-core Caryophyllales supports the sister relationship between Caryophyllales and asterids.