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Article

Genetic Relationships and Diversity of Common Buckwheat Accessions in Bosnia and Herzegovina

by
Jasmin Grahić
1,*,
Arnela Okić
1,
Silvio Šimon
2,
Mirha Djikić
1,
Drena Gadžo
1,
Ivan Pejić
3 and
Fuad Gaši
1
1
Faculty of Agriculture and Food Sciences, University of Sarajevo, Zmaja od Bosne 8, 71 000 Sarajevo, Bosnia and Herzegovina
2
Ministry of Agriculture, Ulica Grada Vukovara 78, 10 000 Zagreb, Croatia
3
Faculty of Agriculture, University of Zagreb, Svetošimunska cesta 25, 10 000 Zagreb, Croatia
*
Author to whom correspondence should be addressed.
Agronomy 2022, 12(11), 2676; https://doi.org/10.3390/agronomy12112676
Submission received: 15 September 2022 / Revised: 7 October 2022 / Accepted: 25 October 2022 / Published: 28 October 2022
(This article belongs to the Section Crop Breeding and Genetics)
Browse Figures
Versions Notes

Abstract

:
Common buckwheat (Fagopyrum esculentum Moench.) has a long history of cultivation in the large, mountainous regions of Bosnia and Herzegovina (B&H). Its commercial production is mainly based on the regionally bred variety ‘Darja’, but numerous landraces are also being grown on a smaller scale. As part of the SEEDNet (Southeast European Network on Plant Genetic Resources) project, these landraces have been collected and stored at the Gene bank of the Faculty of Agriculture and Food Sciences in Sarajevo (FAFS). To assess their utilization value, it was first important to investigate if they represent distinct landraces and to identify their genetic relationships with the most commonly grown varieties in the region (‘Darja’, ‘Goluba’ and ‘Čebelica’). Therefore, the aim of this study was to assess the genetic relationships and diversity of the common buckwheat accessions maintained at the FAFS Gene bank, as well as the value of these accessions for future breeding programs, using microsatellite markers and seventeen quantitative and fifteen qualitative morphological traits. The FCA (Factorial Correspondence Analysis) and AMOVA (Analysis of Molecular Variance) revealed that several accessions represent completely distinct landraces which clearly differentiated from the most commonly grown cultivars ‘Darja’ and ‘Goluba’. Conducted morphological analyses revealed that several of the analyzed landraces hold similar characteristics to the ones observed in ‘Darja’ and ‘Goluba’, while others possess unique traits potentially useful in breeding programs.

1. Introduction

Common buckwheat (Fagopyrum esculentum Moenh.), an annual crop with grain-like seeds, has recently attracted considerable interest among consumers, foremost due to its nutritional properties. This pseudocereal crop is characterized by protein content with well-balanced amino acids [1,2] and high contents of rutin, making it suitable for special diets [3] or enriching functional foods [4]. It is worth noting that buckwheat has been cultivated since ancient times as an emergency crop due to its ability to grow under poor agronomic conditions [5]. This is owing to the fact that from an agronomical perspective, buckwheat is considered a low-input crop with a short growing season and therefore suitable for cultivation in mountainous areas of Asia, Europe, and America [6,7].
F. esculentum is believed to have originated in southwestern China [8], from where it spread both east and west, arriving in Europe by the 15th century if not earlier [9]. The earliest modern-day agriculture records on the cultivation of buckwheat in Bosnia and Herzegovina (B&H) are from the 1970s, which state that buckwheat production was conducted in an area of 9000 hectares within former Yugoslavia (which encompassed B&H). However, according to historical records, buckwheat was already being significantly cultivated in almost all mountainous regions of Bosnia and Herzegovina during the 18th century [10]. It is important to note that, in terms of geography, B&H is mostly mountainous, encompassing the central Dinaric Alps. Regarding buckwheat varieties that were present on the official cultivar list during the Yugoslav era, the most prominent was the variety ‘Darja’ developed in the 1980s, through a breeding program conducted in Slovenia, also a part of Yugoslavia at that time.
Currently, the variety ‘Darja’ is the most cultivated common buckwheat variety in the countries of Southeastern Europe [11,12,13,14]. Aside from ‘Darja’, buckwheat producers from the region of former Yugoslavia (Slovenia, Croatia, Serbia, Montenegro and Bosnia and Herzegovina) tend to use local common buckwheat varieties, such as ‘Goluba’ [11,13]), and even older local varieties such as ‘Čebelica’. It is important to mention that producers generally use farm-saved seeds as sowing material, not taking into account the disturbance of varieties’ genetic integrity due to the cross-pollination of common buckwheat [15]. First insights into the current state of common buckwheat germplasm in Bosnia and Herzegovina were obtained through widespread survey missions undertaken as part of the SEEDNet (Southeast European Network on Plant Genetic Resources) project. The survey, as well as the follow-up collecting missions, revealed a number of local buckwheat populations, with long cultivation history, yet carrying no distinct local name. The mentioned material was thus collected and stored at the Gene bank of the Faculty of Agriculture and Food Sciences (FAFS) in Sarajevo. Considering the long history of buckwheat production in Bosnia and Herzegovina, the detection of numerous local varieties was not unexpected. Namely, Song et al. [16] note that most of the varieties grown are in fact local populations adapted to their environmental conditions through cultivation. These local varieties, often referred to as crop landraces, represent a form of agricultural crop diversity and play a key role in breeding programs and food security of local people in mountainous regions [17]. However, before the value of the collected common buckwheat accessions, maintained at the FAFS Gene bank can be assessed, it is important to investigate if all these accessions represent distinct landraces. In addition, it is necessary to identify their genetic relationships with the most commonly grown varieties in the region (‘Darja’, ‘Goluba’ and ‘Čebelica’). Considering the previously described farmer practices which are at odds with the preservation of the genetic integrity among the buckwheat cultivars and landraces grown in B&H, a low genetic differentiation among the analyzed accessions is expected. In order to assess the relationships among the accessions several tools can be employed, namely: morphological descriptors and biochemical and molecular markers. Microsatellite markers or SSRs (simple sequence repeat) are widely used for the analysis of genotype, due to their abundance, random distribution within the genome, high polymorphism information content (PIC) and stable co-dominance [5]. However, these loci are most often not connected to particular agronomic traits. Therefore, a combination of SSRs and morphological descriptors represents the most encompassing approach in the assessment of genetic resources among crops such as common buckwheat. Microsatellites have previously been successfully used in several common buckwheat germplasm studies [5,6,18,19,20], while the combined approach of SSRs and morphological descriptors is far rarer [21,22]. Therefore, the aim of this study was to assess the genetic relationships and diversity of the common buckwheat accessions maintained at the FAFS Gene bank, as well as the value of these accessions for future breeding programs.

2. Materials and Methods

2.1. SSR Analyses

Seeds from 14 common buckwheat accessions (3 reference varieties and 11 local landraces), analyzed in this study, are currently maintained at the Gene bank of the Faculty of Agriculture and Food Sciences in Sarajevo. The seed material for the three reference common buckwheat varieties within the mentioned collection was obtained from ex situ seed collections in Slovenia (‘Darja’ and ‘Čebelica’) and Montenegro (‘Goluba’). Among the eleven local landraces, five were collected from Central B&H, two from Western, two from Eastern and the remaining two from Northwestern B&H. Analyzed buckwheat accessions were first sown in pots, and afterward, 16 seedlings were sampled from each accession for the purposes of genetic analyses.
Genomic DNA was extracted from green leaves of the collected buckwheat seedlings using peqGOLD plant DNA kit-a (Peqlab) according to the manufacturer’s instructions. Overall, 140 common buckwheat samples with the best genomic DNA quality were selected for genotyping (ranging from 8 to 13 samples per accession). Ten primer pairs (Table S1), previously published by Kishore et al. [23], Ma et al. [6] and Iwata et al. [5], were used for SSR amplifications.
The M13F-tail PCR method [24] was used to measure the size of PCR products. PCR amplification was carried out in the total volume of 11 μL, containing 2.5 μL of genomic DNA (1 ng/μL), 0.065 μL of the specific forward primer (5 μM), 0.32 μL of M13 universal primer (5 μM), 0.32 μL of normal reverse primer (5 μM), 1 μL of 10× PCR buffer, 1 μL of dNTP (2.0 mM), 0.5 μL of Betaine (1 M) and 0.05 μL of Taq polymerase (5 U/μL). A Veriti TM Thermal Cycler was used to perform the PCR amplification of SSR sequences (Applied Biosystems, Foster City, CA, USA) with the following temperature cycling program: initial denaturation at 94 °C for 3 min, 32 cycles of 30 s at 94 °C, 45 s at 50 °C, 1 min at 72 °C, followed by 9 cycles of 30 s at 94 °C, 45 s at 53 °C, 1 min at 72 °C and by a final extension at 72 °C for 10 min. PCR products (1.5 μL) were diluted with ddH20 (1:50), then added to 8.75 μL HiDi and 0.25 μL Genescan 500 LIZ size standard. Detection of PCR products was conducted using an ABI 3130 Genetic Analyzer (Applied Biosystems), and the obtained data were analyzed using the software package GeneMapper 4.0 (Applied Biosystems).

2.2. Morphological Analyses

Each accession, analyzed in our study, was grown on 3 m long and 2 m wide plots. Plots were assigned according to a randomized complete block design with three replications (Figure S1). The field trial was conducted during three growing seasons on the regeneration field of the Gene bank in Butmir, Sarajevo, which is characterized by alluvial soil. Fertilization was carried before sowing and included the use of 500 kg/ha each of N, P and K. The weather conditions were measured daily, and during the three experimental seasons, temperature extremes or drought conditions were not observed (Table S2). No fertilizers and pesticides were used during cultivation.
Seventeen quantitative and fifteen qualitative traits were observed on thirty plants for each accession during all the seasons. The observed traits are part of the descriptors established for Buckwheat [25] and include the following: plant height (cm), number of internodes, number of branches, stem length (cm), stem diameter (cm), thickness of stem tissue (cm), number of leaves, petiole length (cm), leaf blade length (cm), leaf blade width (cm), length of cymes (cm), number of flower clusters per cyme, number of cymes per plant, number of seeds per cyme, seed length (cm), seed width (cm), seed weight (g), cotyledon leaf color, growth and branch shoot habit, stem color, leaf color, leaf blade color, leaf vein color, petiole color, leaf blade shape, compactness of inflorescence, branched inflorescence, the color of inflorescence stalk, flower color, seed shape, seed color and seed surface.

2.3. Biostatistical Analyses

Population genetics software SPAGeDI 1.2 [26] was used in order to investigate allele polymorphism. Cervus 3.0.7 was used to calculate the polymorphism information content [27]. A multivariate analysis, factorial correspondence analysis (FCA) based on allele frequencies was performed using Genetix 4.02 [28]. Analyses of molecular variance [29], based on the stepwise mutation model [30] was performed using GenoType software with 1000 permutations [31].
The Bayesian-model-based cluster procedure within Structure v. 2.2.3 [32] was used to assess the genetic structure within the set of 140 common buckwheat samples. K (unknown) RPPs (reconstructed panmictic populations) were calculated on individuals, testing K (log-likelihood) = 1–10 for all samples assuming that each were from an unknown origin. Ten independent runs were conducted for each K. A burn-in period of 200,000 and 500,000 iterations was applied. Structure Harvester v. 0.6.1 [33], which implements the Evanno method [34], was used to estimate K values for the obtained molecular data. After identifying the most probable K values, runs with maximum likelihood were used to assign individuals to specific clusters [35]. The assignment of a to an RPP was conducted by the probability of membership qI selected at 80% according to a recent molecular study on buckwheat [18].
Based on all 32 descriptors, relationships among the 14 common buckwheat accessions were investigated using a Factorial Analysis of Mixed Data (FAMD) [36], which enabled a joint analysis of data sets containing quantitative as well as qualitative variables. The FAMD analysis was carried out in R package ‘‘FactoMineR’’ v. 2.41, function ‘‘FAMD’’ [37]. The results of the FAMD analyses were then used to create a Factor map in order to visually portray the relationships of the accessions based on their morphological characteristics.
Input data for all statistical software used were prepared using MADC v. 2.0 computer program [38].

3. Results and Discussion

3.1. SSR Polymorphism and Genetic Relationships among the Analyzed Accessions

Seven of the ten primer pairs used managed to generate scorable SSR alleles on all of the 140 samples of F. esculentum. The remaining three primer pairs were therefore discarded from further analyses. A total of 58 alleles were detected with the set of seven SSRs, with the highest number of alleles produced by Fem1303 (16 alleles), and the lowest by GB-FE-043 (3 alleles), resulting in an average of 8.3 alleles per locus (Table 1). Lower values of an average number of alleles per locus have previously been reported by Sabreena et al. [18] (4.57), Song et al. [22] (7.9) as well as Ma et al. [6] (5.9). Slightly higher values for this parameter have been published by Bashir et al. [19] (9), who analyzed 52 genotypes of common buckwheat using 15 SSRs. On the other hand, a much higher average number of alleles per locus (40.60) were obtained on 19 indigenous varieties from Japan, which were represented through 588 individual samples [5]. Considering the fact that cultivation of common buckwheat in Japan goes back to the Jumon era (4600–2000 BC) [39] and that the country itself is located near the buckwheat’s center of origin, high genetic diversity within this germplasm is unsurprising.
The observed heterozygosity (HO) ranged from 0.302 for Fes1094 to 0.911 for Fes1497 (mean = 0.592), while the expected heterozygosity (HE) ranged from 0.270, again for Fes1094, to 0.729 for Fes1497 (mean = 0.551). Very similar results were published by Bashir et al. [19], who reported that the values of HO ranged from 0.31 to 0.92 with an average of 0.58. The mean expected heterozygosity calculated on the 14 analyzed common buckwheat accessions was somewhat higher than the one of 0.531 reported by Song et al. [22]. Again, much higher values were reported by Iwata et al. [5] in the study in which they analyzed Japan’s indigenous varieties (0.819).
Polymorphic information content (PIC) ranged from 0.255 for Fes1094 to 0.691 for Fes1497 with the average of 0.504 for all analyzed loci, which is higher than the average values (0.48) reported by Ma et al. [6] and Song et al. [22]. Higher values were obtained by Bashir et al. [19] and Iwata et al. [5], who reported average PIC values of 0.56 and 0.84, respectively. The difference in the values of the described parameters among our study and the previous ones is most likely due to differences in sample size, the number of utilized markers, as well as the diversity of the analyzed germplasm.
A multivariate approach based on an FCA analysis was used in order to assess the genetic relationships of the analyzed common buckwheat accessions (Figure 1). The analysis revealed a clear differentiation between the reference varieties (‘Goluba’, ‘Darja’ and ‘Čebelica’) and the local landraces of common buckwheat. ‘Goluba’ and ‘Darja’ are positioned closely to each other indicating a degree of genetic similarity between these varieties. This is probably due to the use of a narrow genepool for buckwheat breeding in Slovenia. High similarity between ‘Goluba’ and ‘Darja’ was already reported by Grahić et al. [21] in a study which focused on the genetic relationships of common buckwheat varieties from the Western Balkans. On the other hand, the third reference variety, ‘Čebelica’, is clearly separated from the other two. It is important to note that ‘Čebelica’ is registered as Slovenia’s indigenous common buckwheat variety.
Overall, the analyzed local landraces from Bosnia and Herzegovina clustered closely together with some notable exceptions. Namely, three local landraces, Pop01, Pop09 and Pop14, differentiated clearly from all other analyzed accessions, including the reference varieties. Additionally, Pop05 is positioned apart from the main cluster but also the other divergent samples.
The distribution of analyzed accessions, based on their collection site showed that landraces collected from Central Bosnia and Herzegovina clustered closely together. It is important to note that all five accessions were collected from different farms and, in most cases, different municipalities. On the other hand, three pairs of local landraces collected from Western, Eastern and Northwestern parts of the country were clearly separated.
In order to verify the results of the FCA, analyses of molecular variance (AMOVA) were conducted among the individual samples (Table S3). The results of the AMOVA revealed that there was no significant genetic differentiation between ‘Goluba’ and ‘Darja’ (FCT = 0.037; p < 0.1758). On the other hand, statistically significant differentiation was found between ‘Goluba’ and ‘Čebelica’ (FCT = 0.112; p < 0.0170) and ‘Darja’ and ‘Čebelica’ (FCT = 0.112; p < 0.0170). These results have previously been reported by Grahić et al. [21] in a study that focused on only these three varieties, without a wider context of buckwheat landraces from Bosnia and Herzegovina.
The largest differentiation between accessions (FCT) was detected between ‘Čebelica’ and Pop01, whereby 36.4% of the total genetic variation was attributed to genetic diversity among populations and 63.6% occurred within the populations (p < 0.001). Somewhat smaller, but nonetheless significant, FCT values were calculated between the reference varieties ‘Darja’ and Pop09 (FCT = 0.132; p < 0.009), as well as ‘Darja’ and Pop14 (FCT = 0.188; p < 0.0010). It is important to note that ‘Darja’ was taken as the main reference point in these analyses due to its ubiquitous presence in common buckwheat production throughout Bosnia and Herzegovina. The FCT values ranged from 0.135 to 0.307 when comparing Pop5 to any of the analyzed varieties or most divergent accessions (Pop01, Pop09 and Pop14) with a significant p value in all the comparisons. Accessions Pop2, Pop3, Pop4, Pop8, Pop11, Pop16 and Pop18 formed a single cluster with no statistically significant difference between them. Considering the fact that some of the mentioned accessions were collected in different parts of B&H, one can freely assume that sowing material was exchanged between those regions, and that due to the cross-pollination of common buckwheat and the use of farm-saved seeds, none of the analyzed samples have kept their genetic integrity. The loss of genetic integrity of specific populations or varieties in B&H, whether a consequence of seed exchange, where producers mix the farm-saved seeds with the seeds which they received from other regions in order to have enough sowing material, or the result of cross-pollination, has already been reported earlier by a molecular study [40]. Another argument for questionable genetic integrity among common buckwheat landraces can be seen in the results published by Iwata et al. [5], who reported a significant, but nonetheless very low, differentiation among Japan’s indigenous varieties (2.34% of total variance).
A Bayesian analysis was implemented on 140 samples of common buckwheat in order to investigate the structure of the analyzed accessions. ∆K analyses revealed a maximum value for K = 2. However, none of the samples displayed a probability of membership above 80%, and therefore, all of them were determined to be admixed for K = 2 (Figures S2 and S3). In earlier studies, Structure analysis proved very useful in distinguishing two different species of buckwheat (common and Tartary buckwheat) even with a low number of SSR markers used [18,19,20]. However, the Structure analysis had difficulties in distinguishing individual varieties within the common buckwheat species, most likely due to high levels of genetic diversity within the varieties themselves, probably caused by cross-pollination.
The results of FCA and AMOVA clearly indicated a high level of genetic diversity among local landraces of buckwheat cultivated in B&H, as well as the fact that these accessions diverge from the most commonly grown cultivars (‘Darja’ and ‘Goluba’). This indicates that the analyzed accessions are still unutilized in regional and international breeding programs. Additionally, although the genetic integrity among most of the analyzed local accessions can hardly be viewed as intact, most visibly among landraces collected from Central Bosnia and Herzegovina, there are several landraces that clearly differentiate from landraces and varieties most commonly cultivated in B&H.

3.2. Phenotypic Data on Common Buckwheat Accessions

The data obtained through phenotyping, using seventeen quantitative and fifteen qualitative traits, among the analyzed accessions (3 reference varieties and 11 local landraces) were assessed using factorial analysis of mixed data (FAMD). Based on the average (quantitative) and median (qualitative) values for the used descriptors, a matrix was constructed for FAMD purposes (Table S4). The most important morphological traits of the analyzed accessions are given in Table 2.
The results of the FAMD showed that the cumulative proportion of variance for the first two dimensions was 46.54% and that the discriminatory variables were the ones mostly linked with the plant height, as well as leaf and seed properties.
The grouping of individual accessions within the two multivariate analyses, based on the molecular and morphological data, respectively, differed significantly. Considering that the used markers are located in the regions of the genome not tightly correlated to the functional genes, this discrepancy was expected. The genetically most divergent local landraces of common buckwheat are positioned in close proximity to two of the reference varieties. Namely, Pop09 and Pop14 were positioned very close to ‘Goluba’ and ‘Darja’, respectively, while another genetically divergent landrace (Pop01) was positioned between the two reference varieties. Considering that ‘Goluba’ and ‘Darja’ possess desirable agronomic traits, making them preferable for commercial cultivation, the proximity of individual accessions to these reference points on the Factor map (Figure 2) indicates that these landraces are also characterized by similar traits. This is of particular interest in light of a recent report on ‘Darja’, which was shown to be a high-yielding cultivar, excelling in commercial production even in northern parts of Europe [41]. Important agronomic characteristics of ‘Darja’ include a lower plant height and thicker stem, making it less susceptible to lodging, as well as a high number of seeds per cyme, making it high-yielding. Overall, four different local landraces grouped quite closely to ‘Darja’ (Pop02, Pop03, Pop08 and Pop14).
In terms of morphological traits, one of the most interesting local landraces is Pop01 with relatively low plant height; semi-erect, longer branches; high values for the thickness of stem tissue; semi-compact, unbranched inflorescence; and an average number of large seeds in comparison to other analyzed accessions. Relatively low plant height is a beneficial trait among buckwheat, as high plant height can increase lodging [42], while a shorter height improves the resistance to lodging. Although dwarf genes are considered a useful resource [43], as their use in breeding can be an effective approach to building lodging resistance, Morishita and Tetsuka [44] reported a significant positive correlation between plant height and seed yield in common buckwheat. A better option seems to be cultivars with moderately low plant height, thus allowing for better yields as well as some lodging resistance. It is important to note that five of the eleven local landraces displayed an average plant height below 80 cm, making them somewhat comparable to semidwarf variants. Additionally, thicker stem tissue found in Pop01 is also important in terms of lodging resistance, as stem diameter at the ground is also an important factor affecting lodging [45]. Aside from Pop01, Pop18 also had some very interesting traits, such as very high values for the number of branches and the length of inflorescence as well as the highest values for seed weight and wrinkled seed surface (desirable in seed peeling). Specifically, Pop18 had an average seed weight of 32 g per 1000 seeds. The most recent study on morphological traits among common buckwheat landraces, conducted on Himalayan germplasm, reported that the 1000 seed weight among their accession ranged between 17.42 and 22.64 g [46]. One of the less desirable traits found in Pop18 is its stem color, which was green. Namely, buckwheat plants with dark red stem color have been reported to contain higher rutin contents than plants with red–green or green–red stems [47]. Green stem color in buckwheat may indicate a lack of enzymes needed for the anthocyanin biosynthesis [48]. On the other hand, among the 11 investigated local landraces, five (Pop1, Pop8, Pop9, Pop14 and Pop16) were found to have red stems, with green stems only found among two of these landraces. The indigenous Slovenian variety ‘Čebelica’ proved to be one of the most morphologically divergent due to high values for some undesirable agronomical traits such as seed length and plant height. Overall, several of the analyzed landraces hold similar characteristics to the ones observed in the two of the most commercially significant buckwheat varieties, making them suitable for commercial cultivation.

4. Conclusions

Seven microsatellite markers managed to identify four local B&H landraces that were clearly differentiated from the reference cultivars as well as all other landraces. Tight clustering of the remaining seven local accessions, particularly those collected from sites in Central Bosnia and Herzegovina, is a probable result of seed exchange and production practices at odds with the preservation of the genetic integrity among landraces. An important outcome of the genetic analyses is the significant genetic differentiation between all reference cultivars and investigated landraces, indicating that this local material is as of yet unutilized in regional breeding programs. The morphological analyses revealed that many of the landraces possess desirable traits, such as a plant height below 80 cm, larger seeds and wrinkled seed surface, not found among reference varieties, thus making them very desirable for future breeding programs.

Supplementary Materials

The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/agronomy12112676/s1, Figure S1: Experimental plots set up at the regeneration field of Gene bank of the Faculty of Agriculture and Food Sciences in Sarajevo; Figure S2: Plot of ∆K values from the Structure analysis based on SSR data on 140 common buckwheat samples; Figure S3: Bar plot of the results from Bayesian genetic structure analysis of 140 common buckwheat samples with K = 2; Table S1: Microsatellite (simple sequence repeats—SSR) code and DNA sequences of 10 primer pairs used in the analyses of 14 common buckwheat accessions maintained at the Gene bank of the Faculty of Agriculture and Food Sciences in Sarajevo; Table S2: Average values of weather parameters during the growing seasons; Table S3: Analysis of molecular variance (AMOVA) based on seven simple sequence repeat loci for each of the pairs of analyzed common buckwheat accessions (‘Goluba’, ‘Darja’, ‘Čebelica’, Pop01, Pop05, Pop09, Pop14); Table S4: Average values with the standard error for the 17 quantitative and summary of the 15 qualitative traits measured on 30 plants (per year) from each of the analyzed common buckwheat accessions.

Author Contributions

Conceptualization, J.G. and F.G.; methodology, J.G., F.G., S.Š., M.D. and D.G.; software, J.G.; formal analysis, J.G. and F.G.; investigation, J.G., S.Š. and A.O.; resources, I.P.; data curation, J.G.; writing—original draft preparation, J.G., F.G. and A.O.; writing—review and editing, J.G. and F.G.; visualization, J.G.; supervision, F.G., M.D., D.G. and I.P. All authors have read and agreed to the published version of the manuscript.

Funding

This research received no external funding.

Data Availability Statement

Data are available from the Faculty of Agriculture and Food Sciences, University of Sarajevo, on request.

Acknowledgments

The processing fees for publishing this paper were covered by the Mirsad Kurtovic memorial fund.

Conflicts of Interest

The authors declare no conflict of interest.

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Agronomy 12 02676 g001 550
Figure 1. Multivariate analysis (Factorial Correspondence Analysis-FCA) of simple sequence repeat data for 11 local landraces and three reference varieties of common buckwheat, collected from Central (C), Eastern (E), Western (W) and Northwestern (NW) Bosnia and Herzegovina (centroids shown).
Figure 1. Multivariate analysis (Factorial Correspondence Analysis-FCA) of simple sequence repeat data for 11 local landraces and three reference varieties of common buckwheat, collected from Central (C), Eastern (E), Western (W) and Northwestern (NW) Bosnia and Herzegovina (centroids shown).
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Figure 2. Factor map portraying the relationships of 14 analyzed common buckwheat accessions based on their morphological characteristics.
Figure 2. Factor map portraying the relationships of 14 analyzed common buckwheat accessions based on their morphological characteristics.
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Table
Table 1. Characterization of the 7 microsatellite loci used on 14 common buckwheat accessions.
Table 1. Characterization of the 7 microsatellite loci used on 14 common buckwheat accessions.
LociNumber of AllelesHOHEPIC
GB-FE-0433.00.6980.5260.414
GB-FE-1915.00.3330.3860.348
Fem130316.00.5900.6370.614
Fem18404.00.6820.6320.554
Fes10945.00.3020.2700.255
Fes136813.00.6270.6770.654
Fes149712.00.9110.7290.691
Mean8.30.5920.5510.504
HO—observed heterozygosity; HE—expected heterozygosity; PIC—polymorphic information content.
Table
Table 2. Most important quantitative and qualitative traits of the analyzed common buckwheat accessions.
Table 2. Most important quantitative and qualitative traits of the analyzed common buckwheat accessions.
AccessionPlant HeightStem ColorStem LengthStem DiameterThickness of Stem Tissue
Goluba97.83 ± 6.201pink97.70 ± 6.2280.40 ± 0.0460.10 ± 0.002
Darja83.61 ± 3.086green80.51 ± 3.0630.40 ± 0.0190.16 ± 0.006
Čebelica129.24 ± 2.822red127.80 ± 2.8780.53 ± 0.0350.12 ± 0.005
Pop0183.16 ± 2.262red82.70 ± 2.2930.52 ± 0.0230.20 ± 0.029
Pop0271.05 ± 1.597pink69.08 ± 1.6670.51 ± 0.0210.12 ± 0.011
Pop0385.31 ± 2.098pink82.81 ± 2.1960.51 ± 0.0210.23 ± 0.025
Pop0473.21 ± 2.348green70.78 ± 2.0270.32 ± 0.0220.12 ± 0.014
Pop0586.01 ± 3.030pink84.94 ± 2.9330.38 ± 0.0200.10 ± 0.000
Pop0874.16 ± 2.034red73.61 ± 2.1060.31 ± 0.0250.10 ± 0.005
Pop09101.86 ± 1.854red99.50 ± 2.0040.54 ± 0.0190.11 ± 0.018
Pop1173.4 ± 2.369pink73.40 ± 2.3690.36 ± 0.0340.27 ± 0.030
Pop1477.28 ± 1.815red76.50 ± 1.8330.30 ± 0.0200.23 ± 0.026
Pop16102.61 ± 3.137red103.78 ± 3.2060.51 ± 0.0190.12 ± 0.007
Pop1894.35 ± 3.167green94.83 ± 3.2140.50 ± 0.0200.12 ± 0.008
AccessionNumber of Seeds per CymeSeed ShapeSeed ColorSeed SurfaceSeed Weight
Goluba4.40 ± 0.587ovatebrownsmooth0.026 ± 0.0008
Darja8.77 ± 0.735triangularbrownsmooth0.019 ± 0.0013
Čebelica4.91 ± 0.444triangularbrownsmooth0.023 ± 0.0011
Pop014.55 ± 0.563triangularbrownsmooth0.026 ± 0.0006
Pop027.45 ± 0.671ovatebrownsmooth0.023 ± 0.0014
Pop039.47 ± 1.099triangularbrownsmooth0.018 ± 0.0010
Pop048.70 ± 0.925ovatebrownsmooth0.021 ± 0.0013
Pop058.62 ± 0.908ovatebrownsmooth0.025 ± 0.0012
Pop086.84 ± 0.718triangularbrownsmooth0.022 ± 0.0014
Pop095.75 ± 0.481triangularbrownsmooth0.020 ± 0.0021
Pop115.56 ± 0.363ovateblacksmooth0.019 ± 0.0014
Pop143.78 ± 0.280ovatebrownsmooth0.016 ± 0.0013
Pop164.06 ± 0.250triangularbrownsmooth0.019 ± 0.0013
Pop185.75 ± 0.595ovateblackwrinkled0.032 ± 0.0078
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Grahić, J.; Okić, A.; Šimon, S.; Djikić, M.; Gadžo, D.; Pejić, I.; Gaši, F. Genetic Relationships and Diversity of Common Buckwheat Accessions in Bosnia and Herzegovina. Agronomy 2022, 12, 2676. https://doi.org/10.3390/agronomy12112676

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Grahić J, Okić A, Šimon S, Djikić M, Gadžo D, Pejić I, Gaši F. Genetic Relationships and Diversity of Common Buckwheat Accessions in Bosnia and Herzegovina. Agronomy. 2022; 12(11):2676. https://doi.org/10.3390/agronomy12112676

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Grahić, Jasmin, Arnela Okić, Silvio Šimon, Mirha Djikić, Drena Gadžo, Ivan Pejić, and Fuad Gaši. 2022. "Genetic Relationships and Diversity of Common Buckwheat Accessions in Bosnia and Herzegovina" Agronomy 12, no. 11: 2676. https://doi.org/10.3390/agronomy12112676

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