Genetic variation and species relationships in Himalayan buckwheats as revealed by SDS PAGE of endosperm proteins extracted from single seeds and RAPD based DNA fingerprints

SDS PAGE of endosperm proteins and␣RAPD profiles from different accessions of Himalayan buckwheat were studied to determine their genetic variation and phylogenetic relationships. Comparisons based on Jaccard’s coefficient and UPGMA clustering revealed interrelationships broadly in conformity with conventional treatments. Cluster analysis of the endosperm protein profiles of the selected accessions revealed three broad clusters. A moderate level of intraspecific variability was detected in the endosperm protein profile of different accessions of Fagopyrum esculentum, Moench. Three subgroups were detected in cluster 1. Subgroup 1 included varieties designated as Local, Kamroo local, OC-2 and␣VL-7 which were collected from VPKAS, Almora. Local, Kamroo local and OC-2 showed a similarity coefficient of 1.0 inspite of their being identified as different accessions. VL-7 emerged out separately from the rest of the three accessions. Accessions having winged grains and those having striations on the seed coat formed a 2nd and 3rd subgroup, respectively. IC-13145 which been identified as “F. himalianum”, showed 100%␣similarity in endosperm protein profile with IC-13376 (F. esculentum) and 85–90% similarity which other accessions of F. esculentum. Our results indicate that “F. himalianum” belongs to the esculentum group and should not be regarded as a different species. Cluster 2 included all the accessions of F. tataricum (L.) Gaertn. with Sangla 1, 2, 3, 5, 6, 7 and IC-412863 showing 100% similarity. F. cymosum emerged as a separate group distinct from both esculentum and tataricum. Accessions of F. tataricum and F. cymosum did not show significant intraspecific variation in the SDS PAGE profile of endosperm proteins. Out of the 20 primers used, 3 generated robust, easily interpretable amplified products. While a 1490 bp and a 300 bp RAPD was detected only in F. tataricum, a 1154 bp RAPD was detected in all accessions of F. tataricum except in Shimla B-1. This variety is early maturing and has high seed yields.     

Variability in sesquiterpene lactones from the leaves of yacon (Smallanthus sonchifolius) accessions of different geographic origin

The sesquiterpene lactones (STLs) content of glandular trichomes from the leaves of twenty-five yacon (Smallanthus sonchifolius, Asteraceae) accessions, obtained along a latitudinal gradient from Ecuador to northwest Argentina, was characterized by gas chromatography/mass spectroscopy (GC/MS). While accessions from Ecuador, Bolivia and Argentina proved to be very chemoconsistent, significant variation was found in quantitative composition of STLs from accessions in central Peru, the probable region of origin for the species.     

Uptake, Deposition and Wash Off of Fluoride and Aluminium in Plant Foliage in the Vicinity of an Aluminium Smelter in Norway

Uptake, deposition and wash off of fluoride and aluminium in/on plant foliage were studied in two experiments near an Al smelter by growing different plant species under cover with free wind-flow underneath, and in the open, and by washing the leaves. With an average ambient air concentration of 2.2 μg F m³, the F and Al contents in leaves varied between species from 138 to 665 mg F kg^?1 and from 150 to 1025 mg Al kg^?1when grown under cover. An average of about 60% of the F and nearly 70% of the Al could be washed off covered plants. Uncovered plants had 33–51% lower F content, indicating a considerable precipitation wash off. Precipitation wash off varied with species, probably due to different leaf surface properties. The strong effect of precipitation on plant foliage F has to be taken into consideration when comparing differences in F content between years, localities and species. On average about 40–50% of the F and 55% of the Al could be washed off the leaves of uncovered plants. Washed leaves of plants exposed to precipitation had about 20% lower F content, compared to washed leaves of covered plants, thus indicating leaching of F from leaves exposed to rain. In contrast to fluoride, the Al contents in washed leaves of uncovered plants were about 50–80% higher than in covered washed leaves, thus indicating a significant uptake of Al solved in rainwater through the leaf cuticle.     

欧李种质叶片类黄酮含量及不同组分特征研究

为筛选出欧李叶片类黄酮和9种类黄酮物质中单一物质含量较高的品种,并分析确定具有抗氧化和抑制酪氨酸酶活性的物质,本研究以38份欧李种质基生枝成熟期叶片为试材,利用超高效液相色谱法(UHPLC)测定儿茶素、表儿茶素、甘草素、芦丁、槲皮素、槲皮素-7-O-葡萄糖苷、杨梅素、光甘草定、根皮素活性物质含量,对其抗氧化及抑制酪氨酸酶能力进行分析。结果表明,38份欧李种质叶片中,Y09-14品种类黄酮含量最高,为64.84 mg·g^-1,且类黄酮及其9种组分的变异系数均超过20%,表明其遗传多样性丰富。根据叶片类黄酮含量进行聚类分析,发现70%以上的种质为中类黄酮类型。通过对不同种质欧李叶片类黄酮及不同组分物质含量的测定发现,儿茶素、表儿茶素、芦丁、槲皮素-7-O-葡萄糖苷在38份种质中均能被检测到,且儿茶素含量为6.271~935.295 mg·100 g^-1,极显著高于另外8种物质。4种不同活性物质(儿茶素、表儿茶素、芦丁、槲皮素-7-O-葡萄糖苷)与DPPH清除率呈极显著正相关(P<0.01),表明这种活性物质的抗氧化能力较强;芦丁、槲皮素-7-O-葡萄糖苷、光甘草定与酪氨酸酶抑制率呈正相关。本研究为欧李后期相关物质的检测及提取提供了依据。     

Amaranthus genetic resources in Indonesia: morphological and protein content assessment in comparison with worldwide amaranths

The morphological variations (growth habit, leaf shape, leaf color, height, stem diameter, number of branches and internodes, leaf area, blade ratio, leaf thickness, number of leaves, and total leaf area) and protein content of 53 Indonesian amaranths (Amaranthus spp.), consisting of weedy-, vegetable-, and ornamental-types, were assessed. The extent of variation in the Indonesian Amaranthus accessions were compared with the worldwide collection (26 accessions from United States Department of Agriculture (USDA) and 5 ornamentals from Nepal) in the experimental field of the University of Tsukuba, Japan. The variation in average values of most morphological traits and protein content in the Indonesian accessions were similar to those of the worldwide germplasm, but the important parameters that influence vegetable yield (e.g., number of leaves and stem diameter) were superior in the Indonesian accessions. Protein content showed a positive correlation with the number of leaves, whereas a negative correlation was observed with leaf thickness. The Indonesian accession of A. viridis and A. dubius showed a great potential to be further selected as parental lines for high protein content and number of leaves. The relatively high protein content of the leaves of Celosia may also potentially serve as an alternative protein source in the tropics.     

Analyses of the seed protein contents on the cultivated and wild buckwheat <Emphasis Type="Italic">Fagopyrum esculentum</Emphasis> resources

The seed protein contents of 179 accessions belonging to ten species of genus Fagopyrum were determined by means of the Kjeldahl method. The results indicated that there are significant differences of seed protein content among different accessions within the same species. The average protein content of F. esculentum is 12.94%, ranging from 8.81–18.71%, and the protein content of Sibano is the highest, up to 18.71%, Gantian 1 the lowest with 8.81%. The average protein content of tartary buckwheat is 12.17%, with the range from 7.82–18.94%, and the protein content of cultivated tartary buckwheat Qianku 2 is the highest (18.59%), and that of Ganku 1 (7.82%) the lowest. Moreover, there are significant differences of seed protein content among different buckwheat species and between the different buckwheat types (between common buckwheat and tartary buckwheat, between the diploid and the tetraploid, between the cultivated and the wild, and between the big-achene group and the small-achene group). The average seed protein content of F. giganteum is the highest in all tested species, up to 17.81% with the range of 13.91–21.27%, and that of F. callianthum the lowest, only 8.31%. This study was financed by the Natural Science Foundation of China (30270852, 30471116), Program for New Century Excellent Talents in University (NCET-2004-0913), Guizhou Key International Cooperation Project (Qianke Hewai G Zi 2005#400108), Guizhou Oversea Talent Project (Qianren Xiangmu Zizhu Hetong 2004#02), and Key Project of Students in Guizhou Normal University (GNU-SP-2005-SJXY-A01)     

Extraction of rutin from buckwheat (Fagopyrum esculentumMoench) seeds and determination by capillary electrophoresis.

The content of the flavonoid rutin was determined in different milling fractions of buckwheat seeds and in buckwheat stems, leaves, and flowers. The extraction was performed by using a solvent containing 60% of ethanol and 5% of ammonia in water. The extracts were analyzed by capillary electrophoresis (running buffer of 50 mM borate (pH 9.3), 100 mM sodium dodecyl sulfate; determination at 380 nm). In bran fractions the concentration of rutin was 131-476 ppm, and in flour fractions 19-168 ppm. On average, about 300, 1000, and 46000 ppm of rutin were found in leaves, stems, and flowers, respectively. The results indicate that buckwheat could be an important nutritional source of flavonoids, especially in countries with a low mean daily flavonoid intake.     

Phenotypic and genetic characterization of Kyrgyz fine-leaved Festuca valesiaca germplasm for use in semi-arid,low-maintenance turf applications

Fine-leaved Festuca valesiaca Schleich. ex Gaudin (2n = 2x–4x) is native to heavily-grazed, cold, semi-arid, Asian rangelands. However, its potential for low-maintenance turf applications in the semi-arid western United States and its relatedness to other agriculturally important Festuca species have not been investigated. Therefore, a project was designed to identify F. valesiaca accessions that possess horticultural potential when grown under semi-arid growing conditions and to characterize their relatedness to other Festuca species. In 2008, 12 F. valesiaca accessions originating from Kyrgyzstan and eight US. Festuca and one Lolium cultivar were transplanted as replicated, spaced plants to a field nursery at Blue Creek, Utah. Relative vigor, height, width, total biomass (aboveground dry matter yield), seed weight, and seed number were evaluated between 2009 and 2011. Plant height, width, and total biomass of the F. valesiaca accessions examined were approximately equal to the commercial control, ‘Cascade’ (F. rubra L. subsp. commutata Gaudin; 6x; chewings fescue). Plant vigor and seed weight of F. valesiaca accessions PI 659923, PI 659932, W6 30575, and W6 30588 under semi-arid conditions (~300 mm annual precipitation) were significantly (P < 0.05) greater than ‘Cascade’. Moreover, principal component analysis based on all traits as loading factors indicated that these 12 F. valesiaca accessions were distinct from a majority of the other Festuca accessions examined. These F. valesiaca accessions produced abundant amounts of small seed, and this seed yield was significantly correlated with total aboveground biomass (dry weight; r ²  = 0.84, P < 0.001), plant height (r ²  = 0.58, P < 0.05), and plant vigor (r ²  = 0.83, P < 0.001). Amplified fragment length polymorphism (AFLP) analysis (1,454 polymorphic bands) was used to characterize F. valesiaca relatedness to other economically important Festuca species. The AFLP-based, neighbor-joining analysis differentiated F. valesiaca accessions from US Festuca cultivars examined, except for ‘Durar’ (F. ovina L.; 6x; sheep fescue), to which they had strong genetic affinities. Given their morphological attributes, F. valesiaca PI 659923, W6 30575, PI 659932, and W6 30588 should be considered for use in low-maintenance, semi-arid turf improvement programs in the western US.     

Resistance in natural populations of three wild Lactuca species from Israel to highly virulent Californian isolates of Bremia lactucae

Seedlings of 213 accessions representing 9, 14, and 10 Israeli natural populations of the wild Lactuca serriola, L. saligna, and L. aculeata, respectively, were initially screened for their resistance to a pathotype CAVIII isolate of Bremia lactucae. All 60 L. serriola accessions were susceptible while all 83 accessions of L. saligna were resistant. Out of the 69 L. aculeata accessions, 36 (52.2 %) were resistant. From those resistant accessions, 56 L. saligna and 23 L. aculeata accessions were then tested at the seedling stage for their reaction against five highly virulent isolates originating from California and representing the two current major pathotypes and a novel type of B. lactucae; true leaves of adult plants were also tested with two out of these five isolates. Our study supports previous observations that L. saligna is highly resistant to B. lactucae. However, our results provide additional evidence that L. saligna may not be an absolutely non-host plant for B. lactucae at least at a seedling stage, which is in agreement with other recent data for this species. Sixteen (69.6 %) out of the 23 L. aculeata accessions expressed resistance against all isolates tested, even in seedling stage as well as in true leaves of adult plants. This study is probably the first report of detailed screening of resistance to some B. lactucae isolates in natural populations of L. aculeata. These patterns of resistance reactions show that L. aculeata, a species within the primary lettuce gene pool, should be considered as an attractive source of germplasm for resistance breeding of cultivated lettuce (L. sativa).     

Heavy Metal Content of Some Savanna Plant Species in Relation to Air Pollution

The Sb, Cr, Ni, As, Pb and Cu contents in leaves of Dichrostachys cinerea, Ficus platyphylla, Isoberlinia doka and Securiniga virosa growing around the Kaduna Refinery and Petrochemical Complex (KRPC) were compared with heavy metal contents of the same species growing in a relatively “clean” environment, during the rainy and dry seasons of 2001. Concentrations of Cr, Ni, As and Pb in the leaves of the plant species monitored were significantly lower during the dry season than during the rainy season, particularly in the polluted site (P < 0.05). The concentrations Sb and Cu were lower only in F. platyphylla and S. virosa. Patterns of accumulation of individual heavy metals were variable. All four species of plants accumulated Pb to a higher degree (P < 0.05), than Cr and Ni, which had accumulation patterns similar, and higher than that of As. Arsenic was the least (P < 0.05) accumulated. Accumulation of Cu was different to other metals, being higher than Pb, Cr, Ni, and As (P < 0.05), and second only to Sb, which was hyper-accumulated in all four plants species. Heavy metal contents of the four species varied between rainy and dry seasons. The differences in metal contents observed between seasons, could be linked to relative availability of cations to plants as influenced by soil moisture content. Variations in pattern of accumulation of individual metals, could be related to the genetically induced capacity of plants to alleviate the toxic effects of heavy metals through mechanisms, which include binding metals to peptides and organic acids, formation of phytochelatins, cellular adaptations, efflux pumping systems and other mechanisms.     

Phylogenetic relationships among natural populations of perennial buckwheat, Fagopyrum cymosum Meisn., revealed by allozyme variation

Twenty natural populations of F. cymosum, an insect-pollinated allogamous wild species of Fagopyrum, were investigated for their chromosome number and allozyme variation at 10 Ioci encoding 8 enzymes. Diploid populations were obtained in Sichuan, Yunnan and Tibet in China, whereas tetraploid populations were obtained in Tibet, the Himalayan hills, Thailand and southern China. Both diploid and tetraploid populations maintained a large amount of allozyme variation. The average heterozygosity, H_e, ranged from 0.045 to 0.389 (0.213 on average). Tetraploid populations showed higher H_e values than diploid populations. Natural populations of F. cymosum were locally well differentiated (G_ST = 0.322), probably due to reproductive isolation between the two ploidy levels and to distribution over wide areas from southern China to the Himalayan hills. The phylogenetic tree constructed by the neighbor-joining method based on allozyme variation clarified two distinct groups of diploid populations, the Sichuan and Yunnan groups. As for the tetraploid populations, polyploidization occurred twice independently, once in Yunnan and once in eastern Tibet. Tetraploid F. cymosum diffused to warmer areas such as southern Yunnan and Thailand as well as to cooler areas of Tibet and the high Himalayan hills and became the most widely distributed of the Fagopyrum species.     

Seasonal variation in nutrient concentration in leaves and inflorescences of Avocado

The concentrations of nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), and magnesium (Mg) were determined in leaves from the north and south sides of the tree canopy, and in apical and basal parts of winter and fall inflorescences of 20‐year‐old ‘Colin V‐33’ avocado (Persea americana Mill.) trees, to observe the seasonal variation of all these nutrients in leaves and in inflorescences. The concentration of all the elements evaluated did not change considerably during the months of flowering and vegetative growth, but it changed with leaf age. The concentration of Ca and Mg were 14 and 7% higher, in the leaves on the south side of the canopy than in those on the north side. Inflorescences had higher P and K, and lower Ca and Mg concentrations than leaves. The concentration of Ca and Mg was higher in the winter inflorescences than in the autumn ones. The concentration of P was significantly higher in the apical part of the inflorescence than in the basal part.     

Impact of the Mediterranean Urban Environment on Photosynthetic Efficiency of Quercus ilex Leaves

Environmental constrains and anthropogenic changes can contribute simultaneously to the limitation of plant performance in Mediterranean urban areas. Photosynthetic efficiency of Quercus ilex L. leaves from the urban area of Naples was evaluated by comparison with leaves from remote site (Vesuvius National Park). The photosynthetic performance of leaves with different age (current year [CY], 1- [1Y] and 2-year old [2Y]) was assessed by measurements of gas exchange, chlorophyll fluorescence and pigment contents. The photosynthetic activity at the urban site was higher than that at the remote site for the CY leaves, while the opposite was observed for the 1Y and 2Y leaves. The growth of Q. ilex trees was mainly sustained by the CY leaves at the urban site, whereas at the remote site the photosynthetic activity was not affected by leaf ageing. In the urban environment, Q. ilex leaves strongly decreased the photosynthetic performance with leaf ageing as highlighted by leaf gas exchanges, although the photochemistry did not show any significant change. The mature leaves of urban site drove the reductive power deriving from the photochemistry more in non-assimilative processes rather than in carbon assimilative ones, avoiding a strong decrease of PSII photochemical efficiency.     

Eco-geographical distribution of <Emphasis Type="Italic">Lactuca aculeata</Emphasis> natural populations in northeastern Israel

The wild lettuce, Lactuca aculeata Boiss. et Ky., is closely related and fully interfertile with cultivated lettuce, Lactuca sativa L. Altogether, 155 accessions of this wild species, which is rare conserved within the world’s germplasm collections, were collected from 12 localities throughout northeastern Israel. Ten of these localities represent different sites across the Golan Heights, while two, high-density populations were suprisingly found at the Hula Plain (first report of this species in this region). Lactuca aculeata was recorded at various elevations (222–968 m a.s.l.) and habitats. The taxonomic status of 30 L. aculeata accessions was morphologically validated during ex situ seed regeneration of 31 random accessions representing all 12 localities. Characterization of 12 traits showed that the Israeli populations of L. aculeata do not exhibit broad morphological variability, but more great levels of variation were obtained for developmental traits. Relatively little variability was observed in the morphology of cauline leaves or in the distribution of anthocyanin pigmentation in bracteae. In two cases, L. aculeata plants lacked trichomes in the inflorescence, a typical feature in the majority of plants. These unique collections of wild lettuce may carry novel sources of genetic variation for a wide range of traits and, thus, should be of interest for careful evaluation and exploitation in lettuce breeding.     

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.     

Variability of phenotypic and morphological characteristics of some Lathyrus sativus L. and Lathyrus cicera L. accessions and nutritional traits of their seeds

The object of the study comprised 54 grass pea (Lathyrus sativus) accessions originating from Czech Republic, Hungary, Slovak Republic, Poland, Ukraine and Russia together with 18 red pea (Lathyrus cicera) accessions from Greece, Spain and Italy. The plants were grown under the same conditions in Poland. Some phenological and morphological traits of plants as well as some seed morphological properties and nutritive value were investigated. Lathyrus sativus seeds contained more protein (average 300?g vs. 255?g?kg^?1 dry matter (DM)), whereas the differences in the average contents of the other basic nutrients between both these species as well as between particular accessions were not substantial. Except for a low level of methionine, the protein of both species showed fairly high concentration of other exogenic amino acids (EAAI around 63–64%). Fat of both L. sativus and L. cicera seeds was found to contain a high level of polyunsaturated fatty acids (66.9 and 58.6%, respectively), mainly linoleic (57.4 and 47.2%, respectively). The researched L. sativus accessions had a quite low level of β-ODAP (733?mg with a range of 0.583–1.340?mg?kg^?1 DM). Slightly higher content of this ANF (1,168?mg with 911–1,349?mg?kg ^?1 DM) in L. cicera seeds was noted. Average tannin concentration in the L. sativus and L. cicera accessions reached 3.3 and 6.4?g?kg^?1 DM, respectively. Average content of this ANF in L. sativus significantly correlated with the colour of both, flowers and seeds; with generally, a darker colour associated with higher levels of tannins.