The development of modern approaches to the genetic improvement of the tree crops Ilex paraguariensis (‘yerba mate’) and Ilex dumosa (‘yerba señorita’) is halted by the scarcity of basic genetic information. In this study, we characterized the implementation of low-cost methodologies such as representational difference analysis (RDA), single-strand conformation polymorphisms (SSCP), and reverse and direct dot-blot filter hybridization assays coupled with thorough bioinformatic characterization of sequence data for both species. Also, we estimated the genome size of each species using flow cytometry. This study contributes to the better understanding of the genetic differences between two cultivated species, by generating new quantitative and qualitative genome-level data. Using the RDA technique, we isolated a group of non-coding repetitive sequences, tentatively considered as Ilex-specific, which were 1.21- to 39.62-fold more abundant in the genome of I. paraguariensis. Another group of repetitive DNA sequences involved retrotransposons, which appeared 1.41- to 35.77-fold more abundantly in the genome of I. dumosa. The genomic DNA of each species showed different performances in filter hybridizations: while I. paraguariensis showed a high intraspecific affinity, I. dumosa exhibited a higher affinity for the genome of the former species (i.e. interspecific). These differences could be attributed to the occurrence of homologous but slightly divergent repetitive DNA sequences, highly amplified in the genome of I. paraguariensis but not in the genome of I. dumosa. Additionally, our hybridization outcomes suggest that the genomes of both species have less than 80% similarity. Moreover, for the first time, we report herein a genome size estimate of 1670 Mbp for I. paraguariensis and that of 1848 Mbp for I. dumosa.

Niche modelling software can be used to assess the probability of detecting a population of a plant species at a certain location. In this study, we used the distribution of the wild relatives of lettuce (Lactuca spp.) to investigate the applicability of Maxent species distribution models for collecting missions. Geographic origin data of genebank and herbarium specimens and climatic data of the origin locations were used as input. For Lactuca saligna, we varied the input data by omitting the specimens from different parts of the known distribution area to assess the robustness of the predicted distributions. Furthermore, we examined the accuracy of the modelling by comparing the predicted probabilities of population presence against recent expedition data for the endemic Lactuca georgica and the cosmopolitan Lactuca serriola. We found Maxent to be quite robust in its predictions, although its usefulness was higher for endemic taxa than for more widespread species. The exclusion of occurrence data from the perceived range margins of the species can result in important information about local adaptation to distinct climatic conditions. We discuss the potential for enhanced use of Maxent in germplasm collecting planning.

The aim of the study was to develop the system of polymerase chain reaction (PCR)-based markers for the assessment of genetic diversity and population genetic studies of Gentiana lutea L. as well as to determine the utility of two indices of marker informativeness. The informativeness was determined for 40 PCR primers of different types (random amplified polymorphic DNA, inter simple sequence repeat, inter-retrotransposon amplified polymorphism, resistance gene analog polymorphism and conserved DNA-derived polymorphism markers) by evaluating discriminating power (DL) and resolving power (Rp) in a sample of 30 plants from two populations. Analysis of correlation between the index value and the number of differentiated pairs of genotypes in the given sample revealed that DL is more efficient than Rp; therefore, we selected primers based on the DL value. In total, 12 primers with the largest values of DL were chosen. Analysis of genetic relationship among 86 plants from six populations showed that the number of bands produced by the three of selected primers was sufficient to give average bootstrap support across six key nodes in the dendrogram higher than 85%, while using six of the primers resulted in average bootstrap value exceeding 99%. Thus, a minimal set of three to six selected primers are sufficient for a quick assessment of genetic diversity of G. lutea populations, depending on the sample size and degree of differentiation between populations, while the rest of the primers with DL values above 0.8 may be used for ecogenetic surveys. Preliminary results obtained with selected primers indicate the moderate level of genetic variation within the species and significant differentiation among individual populations.

The capitate glandular trichomes of wild sunflowers (Helianthus spp.) are considered to be effective defence components that act against some herbivorous insects, but cultivated sunflowers are reportedly deficient in glandular trichomes. To investigate whether glandular trichomes have a role in the protection of cultivated sunflowers against insects, in the present study, Helianthus annuus L. accessions were grown to quantify glandular trichome density in wild and cultivated germplasm types and assess potential anti-insect effects of terpenoids in the glandular trichomes of cultivated sunflowers. Evaluation revealed that capitate glandular trichomes are often abundant in cultivated sunflowers; relative to wild H. annuus, inbred maintainer (HA) lines have similar numbers of glandular trichomes per floret, while commercial hybrids have only ≈ 20% fewer trichomes when compared with wild sunflowers. In the laboratory assay, it was found that glandular trichome extracts increased the mortality rates of sunflower moth, Homoeosoma electellum (Hulst), larvae exposed from the neonatal stage to 9 d. In the surviving larvae, the extracts significantly reduced larval mass and head capsule width. Though there are limitations to the value of glandular trichomes for host plant resistance, the feeding deterrent or toxic effects of sesquiterpene lactones and diterpenes in sunflower glandular trichomes are not limited to sunflower moth larvae, suggesting a potential for resistance to other sunflower insect pests. Additional research is required to understand the inheritance and value of glandular trichomes in commercial sunflower germplasm and how the composition of terpenoids in the glandular trichomes of wild H. annuus may differ from that in cultivated material.

Mungbean yellow mosaic virus (MYMV) is a disastrous pathogen of mungbean. It is widespread in most of southern India and no complete resistance has been identified among its commercial cultivars. Two isolates of MYMV, representing its diversity, were used to assess and characterize the susceptibility reaction of all the three species of Vigna. The seeds were agroinoculated with the virus and the presence of the viral DNA was confirmed after 12 d by reverse transcriptase-polymerase chain reaction (RT-PCR) analysis after which the plants were monitored for the expression of symptoms. All of the 20 accessions of Vigna radiata and ten accessions of Vigna mungo were systemically infected with MYMV, and they all produced typical symptoms. On the other hand, the 24 accessions of Vigna umbellata were found to be resistant to both the isolates. For additional affirmation, three representative accessions of V. radiata and V. mungo and all the accessions of V. umbellata were agroinoculated, and quantitative RT-PCR was performed for the quantitative detection of the MYMV. The mRNA transcripts of MYMV were detected in V. radiata and V. mungo plants but not in the V. umbellata plants. Researching the molecular basis of the resistance in V. umbellate against MYMV might definitely be very constructive for developing resistant varieties of mungbean on a commercial scale. This genetic quality offering resistance to MYMV could also be incorporated into V. radiata/V. mungo by means of interspecific crosses.

Crop biofortification is increasingly being recognized as a cost-effective and sustainable approach to address the widespread micronutrient malnutrition arising from Fe and Zn deficiencies. Pearl millet as a cereal crop species has higher Fe density than all other major cereals. Earlier studies in pearl millet have shown that breeding lines, hybrid parents, improved populations and composites having high Fe and Zn densities were often based largely or entirely on iniadi pearl millet germplasm. In an attempt to identify additional sources of high Fe density in this group of germplasm, 297 accessions were screened using Perl's Prussian Blue staining, of which 191 accessions (118 from Togo, 62 from Ghana and 11 from Burkina Faso) were re-evaluated during the 2010 rainy and 2012 summer seasons using the inductively coupled plasma atomic emission spectroscopy method. On the basis of the mean performance over the two seasons (environments), large variability was observed for both Fe (51–121 mg/kg) and Zn (46–87 mg/kg) densities. There was a highly significant and positive correlation between the two micronutrients (r= 0.77, P< 0.01). Of these re-evaluated accessions, 49% had higher Fe density than the high-Fe control commercial cultivar ICTP 8203 (81 mg/kg), and most of these accessions also had Zn density ≥ 61 mg/kg (59 mg/kg for ICTP 8203). A total of 27 accessions (20 from Togo and seven from Ghana) having a Fe density of 95–121 mg/kg (1 standard error of difference above that for ICTP 8203) and a Zn density of 59–87 mg/kg were selected as a valuable germplasm resource for genetic improvement of these two micronutrients in pearl millet.

Since the immediate wild relatives of Sorghum bicolor (L.) Moench are indigenous to Ethiopia, studying their population biology is timely for undertaking conservation measures. A study was conducted to investigate the occurrence of population bottlenecks and to estimate the long-term effective population size (Ne) in wild relatives of sorghum. For this, 40 samples of wild sorghum were collected from two remotely located populations that were allopatric to the cultivated sorghum. The presence of bottlenecks was investigated using heterozygosity excess/deficiency, mode shift and allelic diversity based on nine polymorphic simple sequence repeat (SSR) loci. We also estimated the Ne of the studied populations using two different methods employing SSR mutation models. The expected heterozygosity was found to be 0.41 and 0.71 and allelic richness was 3.0 and 4.9, in Awash and Gibe populations, respectively. Neither the heterozygosity excess nor the mode-shift methods detected signatures of bottlenecks in the studied populations. The effective size of the two wild sorghum populations studied also showed no risk of population reduction in these regions of Ethiopia. Therefore, these allopatric wild sorghum populations can survive by occupying patches by the roadsides and fences, areas within abandoned farm lands, forests, etc., which shows that their wild characteristics of adaptation have been adequate for them to survive from extinction despite extensive deforestation of their habitat for modern agriculture and frequent grazing by livestock. However, this does not guarantee the survival of these species for the future and ex situ conservation measures or policies could help maintain their diversity.

Monitoring and protecting germplasm in genebanks using in situ collections while preserving its original genetic integrity is a priority of germplasm curation. Many germplasm accessions need to be regenerated due to their demand and/or seed condition. The regeneration of wild Helianthus (sunflower) species poses several challenges due to the diversity of 53 wild species. Fatty acid composition of sunflower oil is an important quality factor for the crop. Since oil quality is environmentally influenced, and evaluation of this trait is usually performed on oil from achenes from the original accessions of wild sunflower species, we conducted a study on 72 accessions of eight annual and four perennial taxa of wild sunflower species to compare the oil quality of the original accessions and those regenerated for genebank maintenance. The results showed that the fatty acid composition profiles of achenes from the original and regenerated accessions are not the same. It seems that selection for specific fatty acids in several species will require the analysis of both populations to identify germplasm accessions for use in breeding programmes. It should be borne in mind that accessions of wild species are open-pollinated segregating populations, so one would expect some variability in each succeeding generation. While there may be differences between the original and regenerated accessions, the interrelationships of fatty acids are generally similar in wild and cultivated sunflower species, so there should be no detrimental effects on oil quality when using the wild species for other traits. As more regenerated accessions become available, a more precise relationship between the original and regenerated accessions should emerge.

Core collections are the integral part of biotechnology-aided modern-day crop improvement programmes and utilized for a variety of applications including conventional plant breeding, association mapping, resequencing, among others. Since their advent, determination of core collection size has been based on the size of the whole collection. In this study, we precisely estimated the size of the core collection based on the diversity of the whole collection using the Similarity Elimination method. For each of the elimination cycle, allele retention and pairwise and mean genetic distances were calculated and used as the criteria for the precise estimation of the core collection size. We sampled a coconut core collection with 266 entries by retaining the diversity of the whole collection. During the elimination process, accessions with very rare alleles were eliminated first when compared with those having rare and common alleles. Therefore, our results support the hypothesis that the less frequent alleles seldom contribute to the genetic distance when compared with common alleles. In conclusion, presize can be efficiently utilized in any crop for the precise estimation of core collection size.

This study examined variation in seed zinc (Zn) in a local upland rice germplasm that may affect adaptation in a system of slash-and-burn. Individual seed Zn in farmers' seed lots of local upland rice varieties from a slash-and-burn system was evaluated by staining with dithizone (DTZ). Concentration of Zn in the farmers' seed lots and their single-seed descent genotypes grown at Chiang Mai University was determined by chemical analysis. The DTZ staining of individual seeds in most of the farmers' seed lots covered the intensity of standards that ranged from 19 mg Zn/kg (RD21) to 31 (Nam Roo) mg Zn/kg. Zinc content by chemical analysis was closely correlated with the weighted-average staining for each seed lot. Almost all of the single-seed descent genotypes had higher seed Zn than RD21; two-thirds were higher than Nam Roo. The variation within seed lots detected by DTZ staining was confirmed by seed Zn in the single-seed descent genotypes. Evolutionary adaptation to soil with limited Zn, exacerbated by alkalinity of the ash from slash-and-burn, is made possible by variation in seed Zn among individuals growing together in the same field, and benefits the eaters with Zn-enriched seed.

Centella asiatica (L.) Urban is an important herbaceous medicinal plant with a worldwide distribution. The herb possesses a medicinal value and is used extensively in traditional systems of medicine. The medicinal properties of the herb are attributed to the presence of characteristic triterpenoids and their saponins in the leaves. The major triterpenoids are asiaticoside, madecassoside and their aglycones asiatic acid and madecassic acid, respectively, among others. The present study reports a remarkable qualitative and quantitative variability in secondary metabolites in different accessions of C. asiatica L. as determined by high-performance liquid chromatography (HPLC) analysis. The accessions analyzed in this study can be considered as the core set of discrete chemotypes of C. asiatica. Considerable and contrasting biochemical variations were observed in the terpenoid profiles of the chemotypes. From the basic and applied phytochemical utility, this chemotypic variability in the total content of triterpenoids is important and interesting.

Genebanks are important suppliers of genetic resources to the genomics research community, and access to the resulting information will allow traditional genebank users to better select genetic material for their breeding and scientific programmes. We discuss herein a possible solution to interconnect these data automatically based on semantic web technology.

Microsatellite markers specific to Cheirostylis yunnanensis Rolfe were developed using an enriched genomic DNA library technique. The library was constructed using (AG)20 and (CAG)20 oligonucleotide repeats. A total of 48 primer pairs were designed and tested with 48 C. yunnanensis Rolfe samples, resulting in 11 polymorphic loci. The number of alleles per locus ranged from 2 to 12, with an average of six alleles. The observed and expected heterozygosity ranged from 0.0426 to 0.8085 and 0.0421 to 0.9078, respectively. Of the 11 polymorphic loci, three showed a significant deviation from Hardy–Weinberg equilibrium and one exhibited linkage disequilibrium. Cross-species amplification was tested with five samples of Cheirostylis of unknown species resulting in eight loci that could be amplified, with the number of alleles ranging from one to two. The microsatellite markers developed in this study will be useful for the genetic analysis of C. yunnanensis in order to differentiate species as well as to establish a conservation plan for this species.

Momordica charantia L. (Cucurbitaceae) is a commonly cultivated functional food plant. In the present study, karyological variation among 34 accessions of M. charantia originating from the Indian subcontinent (Bangladesh, India and Pakistan) and Thailand was evaluated. Ploidy levels ranging from diploid (2n= 22) to hexaploid (2n= 66) were observed. Hexaploidy in M. charantia is reported herein for the first time. Chromosome length ranged from 0.8 to 2.9 μm. The hexaploid accessions had submetacentric and subtelocentric chromosomes. All accessions harboured median and submedian chromosomes. Only one hexaploid accession (TOT1140) had chromosomes with subterminal centromeres. There was considerable variation in chromosome length and chromosome arm ratio.

Two unique black pepper accessions with very long spikes, hitherto unreported in the world genepool of black pepper, were collected from the centre of origin of the species and characterized. The two accessions can be a valuable genetic source in the improvement of spike length.

Amphidiploids serve as a bridge for transferring genes from wild species into wheat. In this study, five amphidiploids with AABBUU and AABBNN genomes were produced by spontaneous chromosome doubling of unreduced triploid F1 gametes from crosses between diploid Aegilops (A. umbellulata accessions CIae 29 and PI 226500, and A. uniaristata accession PI 554419) and tetraploid Triticum turgidum (ssp. durum cultivar Langdon and ssp. dicoccum accessions PI 94 668 and PI 349045) species. The composition of high-molecular-weight glutenin subunits (HMW-GS) in these amphidiploids and in their parental A. umbellulata and A. uniaristata species was analysed. As expected, the amphidiploids from T. turgidum ssp. dicoccum accession PI 944668 or PI 349045 and A. umbellulata accession CIae 29 or PI 226500 and A. uniaristata accession PI 554419 showed the same HMW-GS patterns as those of their Aegilops parents, because HMW-GS genes were all silenced in the T. turgidum ssp. dicoccum parents. The amphidiploids from CIae 29 and Langdon inherited all of the HMW-GS genes from their parents except for the Uy type. Using 10 and 15% sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) and 10% urea/SDS–PAGE, 11 Ux and ten Uy types in 16 combinations were observed in 48 A. umbellulata accessions, and two Nx and two Ny types in two combinations were detected in six A. uniaristata accessions. These novel HMW-GS variants may provide new genetic resources for improving the quality of wheat.

Inhibition of α-glucosidase activity is considered an effective means for controlling diabetes by regulating glucose uptake, and blueberries have been shown to possess high levels of inhibitory activity. In the present study, we examined the variations in α-glucosidase inhibition, phenolic and anthocyanin levels, and antioxidant capacity in the peel and pulp of 16 mixed-species rabbiteye hybrids (Vaccinium ashei Reade × Vaccinium spp.), one rabbiteye cultivar (V. ashei) and two highbush hybrids (Vaccinium corymbosum). Peel tissue had, on average, about four times higher levels of α-glucosidase inhibitory activity than pulp, and exhibited significantly higher levels of all other measured activities, even though the peel comprised only a small portion of the fruit. Significant variations in the levels of antioxidant activity were observed; however, no consistent differences were observed between the hybrids with various species composition. Significant positive correlations (r≥ 0.84) were found among α-glucosidase inhibitory activity, total anthocyanin (TA) and phenolic levels, and scavenging activity against ROO∙, ∙OH, 1O2 and H2O2 radicals in the extracts from the peel and pulp. There was a high correlation observed between α-glucosidase inhibitory activity levels and ROO∙(ORAC)peel (r= 0.95). A similarly high correlation with TApeel (r= 0.93) suggests that TA would be a suitable assay proxy if a broader genotypic evaluation of blueberry genotypes is desired.