Diversity of the composition and content of soluble carbohydrates in seeds of the genus <Emphasis Type="Italic">Vicia</Emphasis> (Leguminosae)

Low molecular weight carbohydrates of seeds of 10 species of Vicia, namely: V. angustifolia, V. articulata, V. cordata, V. ervilia, V. johannis, V. macrocarpa, V. monantha, V. narbonensis, V. pannonica and V. sativa were analyzed by the high resolution gas chromatography method. Seeds of the investigated species contain common (glucose, fructose, myo-inositol, sucrose, galactinol, di-galactosyl myo-inositol and raffinose family oligosaccharides—RFOs) and species-specific carbohydrates (d-pinitol and its α-d-galactosides—in V. articulata, V. monantha and V. pannonica or d-ononitol and its galactoside—in V. ervilia). Among the species containing in seeds RFOs as the main α-d-galactosides (V. angustifolia, V. cordata, V. johanensis, V. macrocarpa, V. narbonensis and V. sativa), an additional subgroup can be separated, which contains a set of unknown compounds (found in V. angustifolia, V. cordata and V. macrocarpa). Moreover, several other unidentified carbohydrate-containing compounds were detected exclusively in seeds of V. ervilia. The concentrations of total soluble carbohydrates (TSCs), including sugars, RFOs, cyclitols and galactosyl cyclitols and unknown compounds, in seeds differ significantly (P < 0.05) among the species. RFOs and sucrose are the main fractions in TSCs, with the exception of V. pannonica seeds, containing more galactosyl pinitols, and V. ervilia, in which unknown compounds account for 50% of TSCs. In V. ervilia RFOs occur at the lowest concentration (9.32 mg g^?1 seeds), and the concentration of DGMI (di-galactosyl myo-inositol) is comparable with that of RFOs, which is unusual among all the investigated Vicia species.     

Chromatographic fingerprinting of <Emphasis Type="Italic">Lupinus luteus</Emphasis> L. (Leguminosae) main secondary metabolites: a case of domestication affecting crop variability

Secondary metabolites (SMs), such as alkaloids and raffinose oligosaccharides (RFOs), play important roles in plant physiology. Although alkaloid and RFO phenotypic variation has been reported for yellow lupin (Lupinus luteus L.), most evaluations have used a reduced number of accessions; thus, limiting the understanding of accumulation patterns and variation ranges. The main goal of this research was to assess alkaloid and RFO content in a diverse L. luteus sample to understand possible SM accumulation patterns across this legume species. Eighteen yellow lupin accessions were analyzed using high performance thin layer chromatography to provide insights on seed and leaf RFO and alkaloid phenotypic variation. Co-dominant markers (170) were used to examine genetic relationships among L. luteus accessions and possible accumulation patterns across closely related genotypes. Significant differences were observed for seed and leaf RFOs. Total seed RFO accumulation ranged from 79.738 to 131.079 mg g^?1. Raffinose, stachyose, and verbascose were observed in all genotypes’ seeds, but at different RFO concentrations. Raffinose was the only RFO detected in leaves (2.793–0.4224 mg g^?1). Total alkaloid accumulation ranged from 0.22 to 15.12 and 0.00 to 8.007 mg g^?1 for seeds and leaves, respectively. Lupinine, sparteine, and gramine were observed in seeds and leaves, and showed a wide range of variation. Neighbor-Joining (NJ) analysis showed an apparent pattern of seed alkaloid accumulation, most likely due to domestication events. However, high RFO accumulating accessions were scattered across the NJ tree. Alkaloid and RFO significant phenotypic variation will not only help to understand the roles of these SMs in L. luteus, but also to uncover the genetic basis behind their accumulation.     

Ecological and morphological characterisation of two rare and endemic wild edible <Emphasis Type="Italic">Cucumis</Emphasis> species (Cucurbitaceae) of Western Ghats of India

Cucumis silentvalleyi (Manilal, T. Sabu et P.J. Mathew) Ghebretinsae et Thulin and Cucumis indicus Ghebretinsae et Thulin are two rare narrow endemics of Western Ghats of India. Both have morphological resemblance but distinguishable by a few key characters like shape of the leaf, ovary and fruit, and chromosome number. While both are edible and non-bitter, their distribution is mutually exclusive, but overlapping with Cucumis sativus var. hardwickii (Royle) Alef. (the wild and feral form of C. sativus L.). Interspecific crossability studies indicate that both are related to each other and to a lesser extent to Cucumis setosus Cogn. and Cucumis melo L. Being incompletely described, a comparative account of their morphology, phenology, distribution, ecology and conservation are provided.     

Genetic diversity of cultivated rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) and wild rice (<Emphasis Type="Italic">Oryza rufipogon</Emphasis> Griff.) in Asia,especially in Myanmar,as revealed by organelle markers

Asian rice (Oryza sativa L.) is widely cultivated in Asia, where it has been classified into Indica and Japonica Group, the latter is further classified into Tropical and Temperate Japonica Subgroup. O. rufipogon is believed to be the closest ancestor to O. sativa, but it remains unclear whether the two groups arose from a single ancestor or different ancestors. Therefore, here, we investigated the matrilineal ancestors of O. sativa using markers for organelle (chloroplast and mitochondrial) genomes, and 119 O. sativa landraces, 10 O. glaberrima Steud., 115 O. rufipogon Griff. from Asia, and 39 accessions from other wild rice species with AA genomes. We screened 18 organelle markers developed based on polymorphic loci in the organelle genomes. In addition, we used the open reading frame 100 of a chloroplast marker. The results indicated that O. rufipogon first differentiated into two lineages and then further differentiated into Indica and Japonica Group, respectively. Accessions of O. rufipogon (R-1f and R-2d types) from Myanmar appear to be the closest ancestors of Tropical Japonica Subgroup and Indica Group, respectively. Therefore, these wild strains may have made a strong contribution to the domestication of rice landraces in Myanmar.     

Diversity and differentiation of <Emphasis Type="Italic">Oryza sativa</Emphasis> and <Emphasis Type="Italic">O. rufipogon</Emphasis> in Indonesia

Analysis of the genetic structure of Indonesian Oryza sativa and O. rufipogon using neighbour-joining trees based on single nucleotide polymorphism (SNP) and simple sequence repeat (SSR) markers revealed that O. sativa in Indonesia is separated from O. rufipogon. Accessions of O. sativa in this study were differentiated into two major groups, indica and tropical japonica, excluding some varieties. SSR and SNP markers revealed the high value of differentiation (F _ST) and genetic distance (D) between indica and tropical japonica and we discovered four loci by SNP markers and one locus by SSR markers that play a role in differentiation between indica and tropical japonica. Interestingly, genetic diversity (H) in O. rufipogon was lower than that in O. sativa, however H in O. rufipogon was the highest and H in tropical japonica was the lowest when O. sativa was divided into two groups. Inbreeding coefficient (Fst) showed evidences that gene flow (Nm) between species and within species might be one of the mechanisms related to the diversification and differentiation of Indonesian rice germplasm by asymmetric pattern between species and within O. sativa as revealed by SSR and SNP markers. In addition, we found evidences on stabilizing selection in Indonesian rice germplasm and they might be the reasons why Indonesian rice germplasm did not differentiate due to source location of landrace. However, we found a weak relation between SSR and SNP markers probably due to highly polymorphic in SSR and the different properties of both markers.     

Phenetic and genetic diversity in Indian <Emphasis Type="Italic">Luffa</Emphasis> (Cucurbitaceae) inferred from morphometric,ISSR and DAMD markers

Genetic variations and relationships among cultivated and wild genotypes of five taxa of Indian Luffa were examined using inter simple sequence repeats (ISSR), directed amplification of minisatellite DNA (DAMD) markers and morphological characterization. Morphometric evaluation of 21 discrete characters in 51 representative accessions segregated the five taxa of Luffa in three main clusters: the two wild species (L. echinata, L. graveolens) in the first, and the cultivated L. aegyptiaca (smooth gourd) and L. acutangula (ridged gourd)/L. hermaphrodita (Hermaphrodite luffa) in the second and third clusters, respectively. Cumulative data analysis of 15 ISSR and seven DAMD markers revealed high percentage polymorphism (97.67 %), moderate genetic distance (0.06–0.72, avg. 0.51), and low heterozygosity and Shannon index values (H = 0.15; I = 0.22) across all the 76 genotypes assayed. A UPGMA dendrogram, based on the combined marker data, resolved the five taxa in two main clusters with high bootstrap support. The morphological and molecular trees showed incongruence in the number of main clusters resolved and in the disposition of the wild and cultivated taxa in different sub-clusters. The cluster analyses and PCoA plots revealed a nested grouping of the hermaphrodite luffa within the ridge gourd group. The Bayesian STRUCTURE analysis identified three genetic clusters for the five assumed taxa. Outcrossing test revealed a mixed mating system in Indian Luffa. This is the first ever report on the mating system in Luffa using molecular markers. The study also demonstrates the utility of using more than one DNA marker in the assessment of molecular diversity in a widely cultivated crop genus like Luffa with a narrow genetic base.     

<Emphasis Type="Italic">p</Emphasis>-Coumaric can alter the composition of cucumber rhizosphere microbial communities and induce negative plant-microbial interactions

Phenolics from root exudates or decaying residues are usually referred as autotoxins of several plant species. However, how phenolics affect soil microbial communities and their functional significances are poorly understood. Rhizosphere bacterial and fungal communities from cucumber (Cucumis sativus L.) seedlings treated with p-coumaric acid, an autotoxin of cucumber, were analyzed by high-throughput sequencing of 16S rRNA gene and internal transcribed spacer amplicons. Then, feedback effects of the rhizosphere biota on cucumber seedlings were evaluated by inoculating non-sterilized and sterilized rhizosphere soils to sterilized background soils. p-Coumaric acid decreased the bacterial diversity of rhizosphere but increased fungal diversity and altered the compositions of both the bacterial and fungal communities. p-Coumaric acid increased the relative abundances of microbial taxa with phenol-degrading capability (such as Chaetomium, Humicola, and Mortierella spp.) and microbial taxa which contained plant pathogens (such as Fusarium spp.). However, p-coumaric acid inhibited the relative abundances of Lysobacter, Haliangium, and Gymnoascus spp., whose species can have pathogen-antagonistic and/or plant-growth-promoting effects. The positive effect of cucumber rhizosphere microbiota on cucumber seedling growth was reduced by p-coumaric acid. Overall, our results showed that, besides its direct phytotoxicity, p-coumaric acid can inhibit cucumber seedling growth through generating negative plant-soil microbial interactions.     

Cross-transferability of SSR markers in <Emphasis Type="Italic">Osmanthus</Emphasis>

Developing a molecular tool kit for hybrid breeding of Osmanthus species and related genera is an important step in creating a systematic breeding program for this species. To date, molecular resources have been aimed solely at Osmanthus fragrans with little work to develop markers for other species and cultivars. The objectives of this study were to (1) determine cross-transferability of O. fragrans and Chionanthus retusus derived SSRs in diverse Osmanthus taxa, (2) quantify the influence of locus-specific factors on cross-transferability, and (3) determine the genetic relationships between accessions. We tested 70 SSR markers derived from O. fragrans and C. retusus in 24 accessions of Osmanthus. Sixty-seven markers showed transfer to at least one other Osmanthus species with an overall transfer rate of 84% of loci across taxa. Genotyping with 42 microsatellite markers yielded a total of 367 loci. Number of alleles per locus ranged from 2 to 17 with a mean of 8.7 ± 4.8. Mean observed and expected heterozygosities were 0.560 ± 0.225 and 0.688 ± 0.230, respectively. Percent of polymorphic loci ranged from 40% in Osmanthus delavayi to 100% in O. fragrans. Osmanthus fragrans had the highest mean number of alleles per locus (4.2) while O. delavayi had the lowest (1.1). A reduced suite of eight-markers can distinguish between accessions with non-exclusion probabilities of identity from 3.91E?⁰⁴ to 2.90E?⁰⁷. The SSR markers described herein will be immediately useful to characterize germplasm, identify hybrids, and aid in understanding the level of genetic diversity and relationships within the cultivated germplasm.     

Morphological variation related with environmental factors in endemic and threatened <Emphasis Type="Italic">Jatropha</Emphasis> species of Tehuacan-Cuicatlan,Mexico

Jatropha spp. from Mexico includes high species richness and endemism; five species inhabit in the Tehuacan-Cuicatlan Biosphere Reserve (TCBR), and they are important resources as food, medicine and biofuel. The assessment of morphological and agronomic characteristics is essential to identify, use and maintain plant genetic resources. Given the lack of information on the morphological variability of Jatropha species in relation to environment, the objective was to analyze the influence of physiographic, climatic, and anthropogenic factors in the morphological variability of the species: Jatropha neopauciflora and J. rzedowskii, both not-endangered; J. oaxacana, special protection; J. ciliata and J. rufescens, both endangered in the Reserve. Twelve quantitative morphological variables were measured in 24 populations of these species; 14 environmental variables were registered, and the disturbance index in the sites was estimated. The information was analyzed with Principal Components Analysis (PCA), Cluster Analysis and Canonical Correspondence Analysis (CCA). PCA detected interspecific variation: J. ciliata and J. rufescens have longer and broader leaves and longer flowers, while the other three species have smaller leaves and flowers. J. oaxacana population has intermediate size of leaves, fruits and seeds, compared with those of J. neopauciflora and J. rzedowskii. CCA detected intra-specific variation among the populations of J. neopauciflora and J. rzedowskii, which were separated in two groups due to fruit and seed size. Axis 1 of CCA correlated positively with altitude and annual temperature range, and negatively with mean annual temperature; at the intra-specific level, both species are adapted to variations of temperature and altitude.     

Untapped amaranth (<Emphasis Type="Italic">Amaranthus </Emphasis>spp.) genetic diversity with potential for nutritional enhancement

Significant genetic diversity was observed in 218 out of a total of 1309 accessions of amaranth (Amaranthus hypochondriacus L.) and its seven wild relatives, A. spinosus L., A. dubius Mart. ex Thell., A. hybridus L., A. tricolor L., A. cruentus L., A. caudatus L., A. retroflexus L. for 24 nutritional parameters including total oil content, fatty acid profile, total protein content and amino acid profile. Diversity for total oil content (6.42–12.53%), linoleic acid (25.68–54.34%), oleic acid (21.97–42.01%) of the total fatty acids, total protein content (7.84–18.01%), among important essential amino acids; lysine content (0.66–11.12 g/16 g N), methionine (0.35–4.80 g/16 g N) and half cystine and (0.12–8.32 g/16 g N) was reported. The un-weighted pair-group method using arithmetic average cluster analysis based on pair wise Euclidean genetic distance grouped the accessions into seven major clusters. Histidine, half cystine, tyrosine, essential amino acids, total oil content, linoleic acid and oleic acid content were the major parameters contributing significantly to genetic diversity. Present findings indicate that significant diversity exists for nutritional parameters in amaranth germplasm. The promising accessions with higher multiple nutritive traits; protein content (>16%), oil content (>11%), lysine content (>7.5 g/16 g N) and EAA higher than the FAO reported values, were identified. This is the first report on detailed nutritional analysis of diversity collected worldwide. These could be used as potential breeding material for nutritional enhancement through genetic improvement. This will help in overcoming the “triple burden” of malnourishment, hidden hunger, and obesity.     

Species delimitation,genetic diversity and structure of the European indigenous wild pear (<Emphasis Type="Italic">Pyrus pyraster</Emphasis>) in Saxony,Germany

The present study investigated genetic diversity, structure and hybridization in a collection of the endangered wild pear species Pyrus pyraster (L.) Burgd. A total of 278 putative ‘true type’ P. pyraster trees originating from seven populations in the federal state of Saxony in Germany were analyzed along with 35 pear cultivars commonly cultivated in Saxony. The genetic analysis was performed using nine nuclear microsatellite markers (ncSSR) and two paternally inherited chloroplast marker (cpDNA) amplifying in the intergenic spacer region trnQ–rps16 and the intron region rps16. On basis of the ncSSR dataset after STRUCTURE analysis 80 % of the wild pear individuals were assigned as ‘true type’ P. pyraster genotypes. The cpDNA analysis showed shared haplotypes in P. pyraster and P. communis but with an unequal frequency in both species. The analysis of molecular variance resulted in a moderate (ncSSR) and great (cpDNA) variation among ‘true type’ P. pyraster and the pear cultivars. The genetic diversity in the ‘true type’ P. pyraster populations was still high and the genetic structure between the populations low (ncSSR and cpDNA) indicating a genetic exchange between the populations by pollen and seeds. The clear discrimination between the P. pyraster and P. communis confirms our expectation of the existence of ‘true type’ P. pyraster individuals in the study area. The existing genetic integrity and the high genetic diversity argue for the implementation of preservation measures in P. pyraster.     

Relationships among <Emphasis Type="Italic">Crataegus</Emphasis> accessions sampled from Hatay,Turkey, as assessed by fruit characteristics and RAPD

The genus Crataegus known as hawthorns, is the largest genus among the Maloideae, which comprises 265 species. Turkey is one of the genetic centers of Crataegus and there are more than 20 species found in Turkey. The fruits of Crataegus are used as food and have high flavonoid, vitamin C, glycoside, anthocyanidin, saponin, tannin, and antioxidant levels. In this study, we attempted to characterize 15 Crataegus accessions sampled from Hatay, located in Eastern Mediterranean region of Turkey. The accessions belonged to several species; C. aronia (L.) DC. var. aronia, C. aronia var. dentata Browicz, C. aronia var. minuta Browicz, C. monogyna Jacq. subsp. azarella (Griseb.) Franco, and C. orientalis Pall. ex M. Bieb. var. orientalis. Fruit characteristics of the accessions exhibited considerable variation. The multivariate, principle component and cluster analyses indicated that the accessions belonged to three groups: (1) C. aronia var. arona accessions; (2) C. aronia var. dentata accessions; and, (3) C. monogyna subsp. azarella and C. orientalis var. orientalis accessions. The principle component analysis results also revealed that the first three components explained 46, 21, and 14% of the variation, comprising a total of 81%. The fruit length and width, leaf area, and soluble solids contents were highly correlated characteristics for the first three components. The 19 RAPD primers generated a total of 107 bands, where 76 of these were polymorphic. The molecular data analyses by principle coordinate and clustering showed similar results to those of pomological characteristics. There were three groups, (1) C. aronia var. arona accessions; (2) C. aronia var. dentata accession; and, (3) C. monogyna subsp. azarella. C. orientalis var. orientalis accession grouped with C. aronia var. arona accessions. Therefore, it can be concluded that, overall, the diversity patterns of pomological and molecular data, generated by RAPD, for Crataegus are in good agreement and the accessions of C. aronia var. aronia, C. aronia var. minuta, C. monogyna subsp. azarella and C. orientalis var. orientalis accessions.     

Quantitative and qualitative analysis of alkaloids composition in the seeds of a white lupin (<Emphasis Type="Italic">Lupinus albus</Emphasis> L.) collection

White lupin (Lupinus albus L.) has unexploited potential as a crop plant due to its high seed yield as well as protein and oil content in seeds. Well-characterized collections of gene resources are very important for breeding as a source of genetic variation. This paper presents the results of analyses for total content and qualitative composition of alkaloids in seeds of 367 L. albus accessions from the Polish Genebank. Accessions were divided into four classes of origin: wild collected material, land races, breeding lines, and cultivars. Apart from the expected broad variation as well as strong differentiation in the alkaloid content, a clear influence of domestication was observed. This was shown as an apparent decrease in the alkaloid content in breeding lines and cultivars classes. The total alkaloid content varies from 0.02 to 12.73% of the seed dry weight. Six major alkaloids (abundance >1%) were revealed: lupanine (28.22–94.49%, mean 76.06% in total content), 13-hydroxylupanine (0.10–32.78%, mean 8.23%), multiflorine (0.00–21.67%, mean 5.52%), albine (0.00–18.55%, mean 4.48%), angustifoline (0.24–12.14%, mean 2.07%), 11,12-seco-12,13-didehydromultiflorine (0.00–12.28%, mean 1.74%). Owing to its abundance, lupanine was found to be the most closely correlated to the total alkaloid content.     

Effect of <Emphasis Type="Italic">Eucalyptus</Emphasis> forests on understory vegetation and soil quality

Purpose

Eucalyptus forest plantations are normally devoid of understory vegetation that is often assumed to be associated with Eucalyptus allelopathic effects. The objective of this study was to determine the influence of high soil compaction and low soil moisture content on inhibition of the germination of understory seeds in Eucalyptus forests and thus would result in the scarcity of understory vegetation.

Materials and methods

The soil water content above the depth of 1 m of six major understory vegetation types was analyzed to determine if there was a correlation between soil water content and understory vegetation. The effects of soil treatment (soil-loosening vs. no soil-loosening) and water supply amount (2500, 2000, 1500, 1000, 500, 250, or 0 ml of water per day) on the seed germination rate of Stylosanthes sp. were explored using an artificial climate chamber experiment. Influence of soil source (five Eucalyptus forest soils vs. two non-Eucalyptus forest soils) and water supply (0, 50, 150, 200, or 400 ml of water every day) on the germination rate of five types of seed were assessed using a three-factor analysis of variance (ANOVA).

Results and discussion

Soil-loosening and water supply significantly (P?<?0.05) increased seed germination rate with the contribution rates of 26.14 and 42.93 %, respectively. Analysis of variance for three-factor experiments revealed a significant (P?<?0.05) effect of water supply and vegetation seed type on the germination rate of plant seeds. No significant effect of soil type was observed on germination rate, indicating that germination rate was not affected by soils in Eucalyptus forest.

Conclusions

The conservation of soil characteristics, such as water content and compaction, during the development of a Eucalyptus forest plantation may be an effective strategy for encouraging the growth of understory vegetation. This study highlights the importance that in dry areas or areas prone to long-term drought, it would be preferable to retain native vegetation.

     

Leaf epidermis characters of Iranian <Emphasis Type="Italic">Pyrus</Emphasis> L. (Rosaceae) and their taxonomic implications

Pyrus L. is a taxonomically difficult genus mainly because of interspecific and intergeneric hybridization. A majority of the wild species of Pyrus occurs in Iran, making this region very important for any study of the systematics of the genus. In this investigation, seven quantitative and seven qualitative characters of leaf epidermis, particularly epidermal cells shape, anticlinal walls patterns, type and size of stomata, and cuticular ornamentation of 22 taxa of Pyrus plus Malus orientalis are studied for assessment of their taxonomical value at different levels. Scanning electron microscopy and light microscopy were used for evaluation of epidermis features of these taxa. Some characters such as shape and anticlinal wall pattern of epidermal cells, presence of adaxial stomata, pattern of cuticular ornamentation and epicuticular wax types are useful in separation of some Pyrus taxa. Moreover, Malus orientalis is recognized easily from all Pyrus species via the density of stomata. According to ANOVA analysis, all quantitative variables show high level of between-group differentiation. Also, based on the hierarchical clustering results gained by multiple correspondence analysis and factor analysis of mixed data analyses of the current study, micromorphological characters of epidermal cells reflect the ecological conditions of pear taxa, so most of the mesophyte and xerophyte taxa can be separated from each other in accordance with epidermal cells characters, specially the anticlinal wall pattern. Totally, the application of some other biological markers, particularly suitable molecular data are needed to present well-resolved relationships among these taxa.     

Genetic resources for arcelin,a stored product insect antimetabolic protein from various accessions of pulses of Leguminosae

Pulses, the most important source of high-quality dietary proteins are prone to serious damage due to insect pests, of which 20–25% damage was inflicted by stored product bruchid pest, Callosobruchus maculatus (F.). An alternative approach to control these insect pests apart from fumigation is the use of primary metabolites involved in plant defense mechanism. In the present study, one of such molecule, an insect anti-metabolic protein, arcelin was screened from nine different wild and cultivated varieties of pulses. Insect feeding bioassay with C. maculatus revealed that seven of these seed varieties were not infested by the bruchid insect pest. Additionally, the presence of arcelin polypeptide subunits (27–42 kDa) was demonstrated in the seeds of Phaseolus limensis L. and Phaseolus lunatus L. whereas, an insignificant amount was observed in Lablab purpureus L. Besides, seed kernel extract of P. lunatus registered a highest HA activity against native human-A erythrocytes and this activity was increased to several fold when the erythrocytes were treated with proteolytic enzymes revealing the characteristic feature of arcelin. Similarly, HA activity was observed when rat and mouse erythrocyte was treated with pronase, whereas, no titer value was observed upon treatment with its native erythrocytes. Of the 24 carbohydrates tested, only acetylated amino sugar, N-acetyl-d-galactosamine inhibited the agglutinating activity of crude seed extract against human-A RBC. Among the three glycoproteins tested, none were found potential to competitively inhibit the agglutination activity against human-A erythrocytes. Overall, this study reveals the presence of arcelin, an anti-metabolic insecticidal protein in seeds of P. lunatus.     

Screening species of <Emphasis Type="Italic">Pilocarpus</Emphasis> (Rutaceae) as sources of pilocarpine and other imidazole alkaloids

Plants of the Pilocarpus genus (Rutaceae) are popularly known as jaborandi and are the only source of pilocarpine, an imidazole alkaloid used in eye-drops for the treatment of glaucoma as well as for the stimulation of sweat and lachrymal glands. Alkaloid extracts from leaf samples of seven species of Pilocarpus, from the states of São Paulo and Maranhão in Brazil, were analyzed using HPLC–ESI–MS/MS. The samples contained between 0.88 ± 0.04 and 1.00 ± 0.14% of alkaloids in relation to the dry weight of their leaves, with significant differences in results (P ≤ 0.05) found only between Pilocarpus microphyllus planted in the state of Maranhão and Pilocarpus spicatus, Pilocarpus trachyllophus, Pilocarpus pennatifolius and Pilocarpus jaborandi; as well as between Pilocarpus spicatus and Pilocarpus racemosus. Pilocarpine was not found in P. spicatus, whereas in the other species it ranged from 2.6 ± 0.1 to 70.8 ± 1.2% of total alkaloids. P. microphyllus planted in the state of Maranhão for pilocarpine extraction had the highest total alkaloid content, but it had only 35% of pilocarpine in relation to total alkaloids. Three other species contained more pilocarpine in relation to total alkaloids: P. jaborandi (70.8%), P. racemosus (45.6%) and P. trachyllophus (38.7%); and could be candidates for pilocarpine extraction. Differences in alkaloid content were significant for all these samples (P ≤ 0.05). Imidazole alkaloids were observed and partially characterized based on their retention times and high resolution mass. The seven species analyzed had different imidazole alkaloid profiles, but only one did not present quantifiable pilocarpine contents in its leaves. The Pilocarpus genus shows potential for the prospection of novel alkaloids.     

Genetic diversity of dihydrochalcone content in <Emphasis Type="Italic">Malus</Emphasis> germplasm

Dihydrochalcones, beneficial phenolic compounds, are abundant in Malus Mill. species, particularly in vegetative tissues and seeds. Phloridzin (phloretin 2′-O-glucoside) is the primary dihydrochalcone in most Malus species including cultivated apple, Malus?×?domestica Borkh. A few species contain sieboldin (3-hydroxyphloretin 4′-O-glucoside) or trilobatin (phloretin 4′-O-glucoside) in place of phloridzin, and interspecific hybrids may contain combinations of phloridzin, sieboldin, and trilobatin. Proposed health benefits of phloridzin include anti-cancer, antioxidant, and anti-diabetic properties, suggesting the potential to breed apples for nutritional improvement. Sieboldin and trilobatin are being investigated for nutritional value and unique chemical properties. Although some of the biosynthetic steps of dihydrochalcones are known, little is known about the extent of variation within Malus germplasm. This research explores the genetic diversity of leaf dihydrochalcone content and composition in Malus germplasm. Dihydrochalcone content was measured using high performance liquid chromatography (HPLC) from leaf samples of 377 accessions, representing 50 species and interspecific hybrids from the USDA-Agricultural Research Service (ARS) National Plant Germplasm System Malus collection. Within the accessions sampled, 284 accessions contained phloridzin as the primary dihydrochalcone, one had only trilobatin, two had phloridzin and trilobatin, 36 had sieboldin and trilobatin, and 54 had all three. Leaf phloridzin content ranged from 17.3 to 113.7 mg/g with a heritability of 0.76 across all accessions. Beyond the potential of dihydrochalcones for breeding purposes, dihydrochalcone composition may be indicative of hybridization or species misclassification.     

Phylogenetic relationships within <Emphasis Type="Italic">Lactuca</Emphasis> L. (Asteraceae), including African species,based on chloroplast DNA sequence comparisons

Lettuce (Lactuca sativa L.) belongs to the genus Lactuca L. and is an important vegetable worldwide. Over the past decades, there have been many controversies about the phylogeny of Lactuca species due to their complex and diverse morphological characters and insufficient molecular sampling. In this study we provide the most extensive molecular phylogenetic reconstruction of Lactuca, including African wild species, using two chloroplast genes (ndhF and trnL-F). The sampling covers nearly 40 % of the total endemic African Lactuca species and 34 % of the total Lactuca species. DNA sequences from all the subfamilies of Asteraceae in Genebank and those generated from Lactuca herbarium samples were used to establish the affiliation of Lactuca within Asteracaeae. Based on the subfamily tree, we selected 33 ndhF sequences from 30 species and 79 trnL-F sequences from 48 species to infer relationships within the genus Lactuca using randomized axelerated maximum likelihood and Bayesian inference analyses. Biogeographical, chromosomal and morphological character states were reconstructed over the Bayesian tree topology. We conclude that Lactuca contains two distinct phylogenetic clades—the crop clade and the Pterocypsela clade. Other North American, Asian and widespread species either form smaller clades or mix with the Melanoseris species. The newly sampled African endemic species probably should be treated as a new genus.     

Genetic relationship of diploid wheat (<Emphasis Type="Italic">Triticum</Emphasis> spp.) species assessed by SSR markers

Genetic diversity of 139 accessions of diploid Triticum species including Triticum urartu, Triticum boeoticum and Triticum monococcum was studied using 11 SSR (simple sequence repeats) markers. A total of 111 alleles with an average of 10 alleles per locus were detected. The polymorphism information content (PIC) of each SSR marker ranged from 0.30 to 0.90 with an average value of 0.62. Among the three Triticum species T. urartu had the highest number of total alleles (Na?=?81), private alleles (Npa?=?15) and showed higher genetic diversity (Hex?=?0.58; PIC?=?0.54). The genotypes from Turkey exhibited the highest genetic diversity (PIC?=?0.6), while the least diversity was observed among 4 Georgian accessions (PIC?=?0.11). Cluster analysis was able to distinguish 139 wheat accessions at the species level. The highest genetic similarity (GS) was noted between T. boeticum and T. monococcum (GS?=?0.84), and the lowest between T. urartu and T. monococcum (GS?=?0.46). The grouping pattern of the PCoA analysis corresponded with cluster analysis. No significant differences were found in clustering of T. urartu and T. monococcum accessions with respect to their geographic regions, while within T. boeoticum species, accessions from Iran were somewhat associated with their geographical origin and clustered as a close and separate group. The results from our study demonstrated that SSR markers were good enough for further genetic diversity analysis in einkorn wheat species.