Genetic structure of Lima bean (<Emphasis Type="Italic">Phaseolus lunatus</Emphasis> L.) landraces grown in the Mayan area

Lima bean (Phaseolus lunatus L.) is an important crop in the Mayan culture. The Mayan area, considered as a main center of Mesoamerican diversity, has been divided into two subareas: the Mayan lowlands and the Mayan highlands. The Yucatan Peninsula is part of the Mayan lowlands and holds the highest number of Lima bean landraces of Mexico, but Lima beans are in high risk of genetic erosion due to intensification of the traditional Mayan agriculture. However, information on genetic diversity of Lima beans of the Mayan highlands is lacking. By using 46 landraces collected in the Mayan area (23 from each subarea) and 73 ISSR loci (inter-simple sequence repeats), we analyzed the structure, diversity and genetic relationships of Lima beans of this part of Mesoamerica. High levels of diversity (H _BAY = 0.45) and genetic structure (F _ST = 0.66) were found for the whole Mayan area. Genetic diversity in the Mayan lowlands was apparently higher than the Mayan highlands (H _BAY = 0.44 and 0.36, respectively); but differences were not statistically significant. Genetic structure between the subareas was high (AMOVA = 30% of total variation), most landraces grouping according to their geographic origin. This study shows the importance of the Mayan culture in the diversification and conservation of Lima beans. The results provide important information that should be considered when implementing strategies to collect Lima bean landraces and planning in situ and ex situ programs to conserve these landraces in the Mayan region.     

Genetic diversity of a <Emphasis Type="Italic">Coffea</Emphasis> Germplasm Collection assessed by RAPD markers

Genetic diversity and relationships within and among nine species of Coffea, one species of Psilanthus and the Piatã hybrid from the Coffee Germplasm Collection of Instituto Agronômico de Campinas (IAC), Brazil were assessed using RAPD markers. Genetic diversity and relationships were evaluated by proportion of polymorphic loci (P), Shannon’s genetic index (H′ and G′_ST) and clustering analysis. The overall RAPD variation among all accessions was mostly partitioned between rather than within species. However, C. canephora and C. liberica showed a high genetic diversity within the species (\({\underline{\hbox{H}'}} \) _sp = 0.414 and \({\underline{\hbox{H}'}} \) _sp = 0.380, respectively) and this was highly structured (high \({\underline{\hbox{G}'}} \) _ST). Genetic diversity from C. congensis and C. arabica was also structured, but with lower levels of genetic diversity (\({\underline{\hbox{H}'}} \) _sp = 0.218 and \({\underline{\hbox{H}'}} \) _sp = 0.126, respectively). The results were consistent with agronomic and molecular studies and demonstrated that the IAC Coffea Collection is representative of the phylogenetic structure observed in the genera. This study devises sampling strategies for coffee germplasm collections and provides genetic diversity parameters for future comparisons among them.     

Morphological and genetic diversity of shea tree (<Emphasis Type="Italic">Vitellaria paradoxa</Emphasis>) in the savannah regions of Ghana

Vitellaria paradoxa C. F. Gaertn., commonly known as shea tree or Vitellaria, is ranked the most important tree species of the savannah regions in the most African countries due to its ecological and economic importance for livelihoods and national economies. However, the savannah regions are the most vulnerable areas to the global climate change. Moreover, the Vitellaria populations on farmlands are threatened by the dominance of old trees with low or lack of regeneration. In this study both morphological and genetic diversity were assessed using several phenotypic traits and 10 microsatellite markers, respectively, to assess the impact of land use and agro-ecozone types on Vitellaria in Ghana. The land use types were forests and farmlands, and the agro-ecozone types included the Transitional, Guinea, and Sudan savannah zones. The mean values of morphological traits, such as diameter at breast height (DBH) and canopy diameter (CD), were statistically different between forest (DBH = 22.20, CD = 5.37) and farmland (DBH = 39.85 CD = 7.49) populations (P < 0.00001). The Sudan savannah zone with mean petiole length of 4.96 cm showed significant difference from the other zones, likely as a result of adaptation to drier climate conditions. Genetic data analysis was based on 10 microsatellite markers and revealed high genetic diversity of Vitellaria in Ghana: mean expected heterozygosity, H _e was 0.667, and allelic richness, measured as number of effective alleles A _e , was 4.066. Both farmlands and forests were very diverse indicating lack of negative influence of farmer’s selection on genetic diversity. Fixation index was positive for all populations (mean F _IS = 0.136) with farmlands recording relatively higher values than forests in all ecological zone types studied, probably indicating less gene flow in the farmlands. Moderate differentiation (F′ _ST = 0.113) was comparable to other similar tree species. Both land use and ecological zone types influenced genetic differentiation of Vitellaria at varying levels. The species was spatially structured across three ecozones and following climatic gradient. The forest reserves are used in situ conservation for Vitellaria in Ghana. High diversity observed in the most arid zones provides opportunity to find and use appropriate plant materials for breeding climate change resilient trees.     

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.     

Dominant Characteristics Between <Emphasis Type="Italic">Microcystis aeruginosa</Emphasis> and <Emphasis Type="Italic">Cyclotella</Emphasis> Sp. Accompanying Dilution Process in Eutrophic Lake

Although dilution of lake water has been used for improvement of water quality and algal blooms control, it has not necessarily succeeded to suppress the blooms. We hypothesized that the disappearance of algal blooms by dilution could be explained by flow regime, nutrient concentrations, and their interaction. This study investigated the effects of daily renewal rate (d), nitrogen (N) and phosphorus (P) concentration, and their interaction on the domination between Microcystis aeruginosa and Cyclotella sp. through a monoxenic culture experiment. The simulation model as functions of the N:P mass ratio and dilution rate (D) (calculated from d) was constructed, and the dominant characteristics of both species were predicted based on the model using parameters obtained in a monoculture experiment and our previous study. Results of monoxenic culture experiment revealed that M. aeruginosa dominated in all conditions (d = 5 or 15%; N = 1.0 or 2.5 or 5.0 mg-N L^?1; P = 0.1 or 0.5 mg-P L^?1) and the predicted cell densities were substantially correspondent to experimental data. Under various N:P ratios and D values, characteristics of domination for each species were predicted, indicating that Cyclotella sp. tended to be dominant under high P concentrations (P ≥ 0.36 mg-P L^?1) when the N:P ratio was less than 7.0, and M. aeruginosa could not form algal blooms at the N:P ratio ≤ 7.0 (N ≤ 0.7 mg-N L^?1). It was also suggested that the dilution rate leading to the Cyclotella sp. domination required 0.20 day^?1 or higher regardless of the N:P ratios.

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Graphical Abstract ? M. aeruginosa and Cyclotella sp. could be a superior competitor in nutrient-limited and nutrient-rich conditions, respectively. ? The simulation model in this study indicated that the predicted cell density and nutrient concentration were substantially correspondent to experimental data. ? The model predicted that Cyclotella sp. tended to be dominant at the P ≥ 0.36 mg-P L^?1 when the N:P ratio was less than 7.0, and M. aeruginosa could not form algal blooms at the N:P ratio ≤ 7.0 (N ≤ 0.7 mg-N L^?1).

     

Genetic diversity and relationships of wild and cultivated <Emphasis Type="Italic">Zanthoxylum</Emphasis> germplasms based on sequence-related amplified polymorphism (SRAP) markers

The genus Zanthoxylum, belonging to Rutaceae, has a long history of cultivation both for economic and chemical values in China. To effectively conserve and sustainably utilize this genus resource, a study on genetic diversity and relationships of Zanthoxylum germplasms was carried out by employing SRAP markers. We used 16 primer combinations to assess genetic variations and relationships among 175 accessions from eight cultivated provenances, including Shandong, Henan, Shanxi, Shaanxi, Gansu, Sichuan, Guizhou and Yunnan. A total of 145 clear repetitive and intense bands were yielded, and the percentage of polymorphic bands was 100 % for per primer combination, indicating a relatively high diversity among Zanthoxylum germplasms. From a geographic perspective, the highest genetic diversity level was observed within Guizhou provenance (N _a  = 1.97, Ne = 1.52, H = 0.31, I = 0.46) while Henan provenance had the lowest genetic diversity (N _a  = 1.68, Ne = 1.45, H = 0.25, I = 0.37). Based on AMOVA results, the abundant genetic variation was mainly caused by variation of intra-provenances (84.96 %), rather than among provenances (15.038 %). The results indicated low genetic differentiation (G _st  = 0.133) and high gene flow (N _m  = 3.2605) among provenances. The neighbor-joining tree revealed that the 175 accessions could be divided into four groups, and groupings indicated a divergence between the cultivated accessions of Zanthoxylum bungeanum Maxim. and Z. armatum DC. Moreover, three accessions of Z. piperitum DC. var. inerme without prickles introduced from Japan gathered one cluster. Cluster IV is composed of accessions of different geographical origin, including 11 wild species and 10 cultivated accessions of Z. bungeanum. The cluster analysis also reflected a relatively close relationship between the geographical origins and the classification of accessions in cluster I. Structure analysis indicated that collected Zanthoxylum accessions could be divided into two major groups. The information obtained from our research would benefit to make use of Zanthoxylum germplasms and assist the management of a Zanthoxylum germplasms collection.     

A Comparative Study on Removal Efficiency of Cr(VI) in Aqueous Solution by <Emphasis Type="Italic">Fusarium</Emphasis> sp. and <Emphasis Type="Italic">Myrothecium</Emphasis> sp.

Environmental pollution with chromium is due to residues of several industrial processes. Bioremediation is an alternative actually considered to remove Cr (VI) from the environment, using adapted organisms that grow in contaminated places. Have been conducted studies with fungi mechanisms of interaction with chromium, most of which have focused on processes biosorption, characterized it by passive binding of metal components of the cell surface, and bioaccumulation, wherein the metal entry to cells occurs with energy expenditure. The paper presents the results of studies carried out on sorption of chromium (VI) ions from aqueous solutions by Fusarium sp. and Myrothecium sp. Both biomasses have the ability to take up hexavalent chromium during the stationary phase of growth and as well inactive conditions. Fusarium sp. showed 26% of biosorption with active biomass and 64% in inactive biomass; meanwhile, Myrothecium sp. obtained 97 and 82%, respectively. Both fungi showed adjust to pseudo-second-order model in active (Fusarium sp. R ² = 0.99; Myrothecium sp. R ² = 0.96) and inactive biomass assay (Fusarium sp. R ² = 0.99; Myrothecium sp. R ² = 0.99). The data of the active biomass test also confirmed to the intraparticle diffusion model (Fusarium sp. R ² = 0.98; Myrothecium sp. R ² = 0.93). The results obtained through this investigation indicate the possibility of treating waste effluents containing hexavalent chromium using Fusarium sp. and Myrothecium sp.     

<Emphasis Type="Italic">Trichoderma</Emphasis> improves the growth of <Emphasis Type="Italic">Leymus chinensis</Emphasis>

Bio-fertilizer application has been proposed as a strategy for enhancing soil fertility, regulating soil microflora composition, and improving crop yields, and it has been widely applied in the agricultural yields. However, the application of bio-fertilizer in grassland has been poorly studied. We conducted in situ and pot experiments to investigate the practical effects of different fertilization regimes on Leymus chinensis growth, with a focus on the potential microecological mechanisms underlying the responses of soil microbial composition. L. chinensis biomass was significantly (P?<?0.05) increased by treatment with 6000 kg ha^?1 of Trichoderma bio-fertilizer compared with other treatments. We found a positive (R² =?0.6274, P <?0.001) correlation between bacterial alpha diversity and L. chinensis biomass. Hierarchical cluster analysis and nonmetric multidimensional scaling (NMDS) revealed that soil bacterial and fungal community compositions were all separated according to the fertilization regime used. The relative abundance of the most beneficial genera in bio-fertilizer (BOF) (6000 kg ha^?1Trichoderma bio-fertilizer) was significantly higher than in organic fertilizer (OF) (6000 kg ha^?1 organic fertilizer) or in CK (non-amend fertilizer), there the potential pathogenic genera were reduced. There were significant negative (P?<?0.05) correlations between L. chinensis biomass and the relative abundance of several potential pathogenic genera. However, the relative abundance of most beneficial genera were significantly (P?<?0.05) positively correlated with L. chinensis biomass. Soil properties had different effects on these beneficial and on these pathogenic genera, further influencing L. chinensis biomass.     

C-banded karyotype of <Emphasis Type="Italic">Thinopyrum bessarabicum</Emphasis> and identification of its chromosomes in wheat background

C-banded pattern in two accessions of Thinopyrum bessarabicum (Save ex Rayss) A. Löve (2n = 2x = 14, E^bE^b) and their idiogram was established. C-banding analysis was further used to identify the chromosomes of Tritipyrum amphiploid (2n = 6x = 42, AABBE^bE^b) and a BC₁F₂ genotype from wheat and Tritipyrum. Two 18S-26S rDNA loci were detected on Th. bessarabicum chromosomes by in situ hybridization using an 18S-26S rDNA probe. E^b chromosomes in Tritipyrum generally were identified by their distinctive C-banding patterns which reflected heterochromatin regions. C-banding procedure resulted in sharp and distinct bands in one or both ends of E^b chromosomes without interval bands. Observed C-bands in E^b genome mainly reflected the telomeric and subtelomeric sequences which also showed more strong signals in genomic in situ hybridization. Results showed the importance of the C-banding technique as a screening tool in identification of addition and substitution lines in the progenies of wheat and Tritipyrum crosses during segregating generations.     

Phytoremediation Potential of <Emphasis Type="Italic">Helianthus annuus</Emphasis> and <Emphasis Type="Italic">Hydrangea paniculata</Emphasis> in Copper and Lead-Contaminated Soil

This study was conducted to assess the hyperaccumulation and phytoremediation potential of copper (Cu) and lead (Pb) in Hardy ‘Limelight’ Hydrangea (Hydrangea paniculata) and the common sunflower (Helianthus annuus). The study also investigated the capacity of these two plants to transpire the metals in a temperature-controlled greenhouse. Plants were grown for 4 weeks and periodically watered with known elemental concentrations of copper oxide nanoparticles, copper sulfate, and lead nitrate. Both H. annuus and H. paniculata accumulated significant amounts of Cu and Pb to be classified as hyperaccumulator species. H. annuus took up significant amounts of Cu in the shoots, specifically the leaves (Cu max.?=?1368 ppm), and easily translocated it from stem to leaf (translocation factor (TF) ranged from 2.7 to 81.0). Pb was not as easily taken up and translocated (TF?=?0.6) as Cu was by this species. H. paniculata took up Cu and Pb in high concentrations but preferentially stored more metals in the stems (Cu max.?=?1757 ppm; Pb max.?=?780 ppm) than in the leaves (Cu max.?=?126 ppm; Pb max.?=?35 ppm). The translocation ability of H. paniculata was much lower for both metals compared to H. annuus. Both Cu and Pb transpired from H. annuus at concentrations of 0.04 and 0.005 ppm, respectively.     

High Zn concentration does not impair biomass,cutting radial growth,and photosynthetic activity traits in <Emphasis Type="Italic">Populus alba</Emphasis> L.

Purpose

Evaluate the efficiency of Populus alba clone Villafranca in the uptake and translocation of Zn from contaminated soils.

Materials and methods

The effects of 48 days of zinc treatment (Zn _t ) on the growth and the photosynthetic activities of P. alba L. clone Villafranca were studied using ZnSO₄ (375 ppm per unit of soil dry weight) added in sand and peat moss substrate at the beginning of the treatment (T ₀) and again after 30 days (T ₁) in order to reach a target Zn concentration of 375 ppm at T ₀ and 750 ppm at T ₁ per unit of soil dry weight.

Results and discussion

Zn uptake in the different organs was analyzed after 30 (T ₁) and 48 days (T ₂) from the beginning of treatment, showing the following order: root ? leaves ≥ woody cutting = stem. The leaf area increased by 12 % in comparison to control plants at the end of second treatment (48 days). Cutting radial growth showed a high synchronicity in the growth rate fluctuation among control and Zn _t plants, but a higher increase in radial diameter of Zn _t cutting was observed starting from day 38 (after 8 days of second Zn _t ) reaching after 48 days 38 % higher than control plants.

Conclusions

Although our data of leaf Zn concentration were in the range usually reported as toxic for plants, Villafranca clone in Zn _t substrate were unaffected in terms of net CO₂ assimilation and stomatal conductance to water vapor.

     

Domestication and saponins contents in a gradient of management intensity of agaves: <Emphasis Type="Italic">Agave cupreata</Emphasis>, <Emphasis Type="Italic">A. inaequidens</Emphasis> and <Emphasis Type="Italic">A. hookeri</Emphasis> in central Mexico

Saponins occur in numerous plants, including agaves, determining benefic and harmful properties to humans; their presence may favor using plants as soap and other products, but also they may cause caustic effects producing contact dermatitis. In domestication, favorable and unfavorable properties of saponins may cause an increase or decrease of their content, respectively. This study quantified and identified saponins among wild and managed populations of three agave species: A. cupreata Trel. et Berger, A. inaequidens Koch with wild and cultivated populations used for mescal production, and A. hookeri Jacobi, existing exclusively cultivated, used for production of the fermented beverage pulque. We studied 272 plants from 19 populations, quantifying contents of crude saponins through spectrometry. In 12 populations, the saponins types were identified by High Performance Liquid Chromatography–Mass-Spectrography-Time-of-Flight HPLC-MS-TOF. The highest crude saponins content was recorded in A. hookeri (26.09 mg/g), followed by A. cupreata (19.85 and 15.17 mg/g in wild and cultivated populations, respectively). For A. inaequidens, we recorded 14.21, 12.95, and 10.48 mg/g in wild, silvicultural managed and cultivated populations, respectively. We identified 18 saponins types, A. inaequidens showing all of them. A hecogenin glycoside (HG1) is found in high amounts in A. hookeri but in low quantities in A. inaequidens and A. cupreata. A. inaequidens had the greatest diversity of saponins. The contents of crude saponins in A. inaequidens and A. cupreata decrease with management intensity, but contrarily to what we expected, it was the highest in A. hookeri. We hypothesize that such high amount could be due to some saponins, probably HG1, may be precursors of sugars.     

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.     

Geographical distribution of wild <Emphasis Type="Italic">Daucus</Emphasis> species in the natural grasslands of the Argentinian pampas

Three wild carrot species have been reported in the argentinian flora: Daucus montanus Humb. et Bonpl. ex Schult., D. montevidensis Link ex Sprengel and D. pusillus Michx. There is a discrepancy among authors about the distinctive morphological traits of the last two species; thus, it is difficult to ascertain if they are truly two distinct taxa. Based on the available literature and in the search of a paradigmatic site, four collection trips were carried out in 2004 and 2005 in Buenos Aires and Southern Entre Ríos provinces. Populations were sampled at 30 sites, and local environmental parameters and associated plant species were recorded. Morphological observations and chromosome counts were carried out on 10 plants/population. Three morphological phenotypes were distinguished: one in 18 populations, all with 2n = 2x = 18, and two in the remaining 12, with 2n = 2x = 22 or aneusomaty (2n = 2x = 20, 22). Populations of the first phenotype were assigned to wild D. carota and the rest, tentatively, to D. pusillus (D. montevidensis?), till further evaluations are carried out in test sites to verify this tentative conclusion.     

Comparison of morphological traits and molecular markers to analyse diversity and structure of alfalfa (<Emphasis Type="Italic">Medicago sativa</Emphasis> L.) cultivars

Diversity and structure of populations may differ substantially between morphological traits and molecular markers. Explanations of such discrepancies are crucial for further progress in breeding as well as for the maintenance of genetic resources. Our objective was to compare indices of among-cultivars differentiation for morphological traits (Q_ST) and molecular markers (F_ST) in alfalfa (Medicago sativa), a legume forage species. Ten cultivars representing the Northern and Southern types commonly grown in Europe were investigated. For each cultivar, 40 genotypes were analysed with 16 SSR markers and four morphological traits measured in two locations and several cuts. Q_ST values were in general high (0.02–0.39) compared to the differentiation observed with molecular markers (F_ST = 0.01), especially for growth habit, indicating that morphological traits were more efficient to structure the diversity than molecular markers. For morphological traits, a clear separation of Northern and Southern cultivars was observed, whereas for molecular markers, no clear structure was detected. Nevertheless, the grazing type cultivar Luzelle was separated from the rest of the cultivars for both morphological traits and molecular markers. Although pairwise differences between cultivars were significant for both morphological traits and molecular markers, the main part of the variation was found within cultivars. This large within-cultivar variation may be explained, besides the outcrossing reproductive mode and autotetraploid genetics, by the recent history of M. sativa domestication in Europe and the frequent seed exchanges. Selection for morphological traits (Q_ST > F_ST) was achieved without modification of within-cultivar neutral diversity.     

Assessment of artificial selection in maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.) and Asian rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) using QTL data

Maize (Zea mays L.) and Asian rice (Oryza sativa L.), two most important cereals for human nutrition, have undergone strong artificial selection during a long period of time. Currently, a number of genes with stronger signals of selection have been identified through combining genomic and population genetic approach, but research on artificial selection of maize and Asian rice is scarcely done from the perspective of phenotypic difference of a number of agronomic traits. In this study, such an investigation was carried out on the basis of 179 published studies about phenotypic quantitative trait locus (QTL) mapping of Zea and Oryza species via QTL sign test. At the overall level, the proportions of antagonistic QTLs of Zea and Oryza species were 0.2446 and 0.2382 respectively, deviating significantly from neutrality. It indicated that these two genera have undergone similar selection strength during their evolutionary process. A previous study showed that 4 traits undergoing the directional selection during domestication were identified in Asian rice via QTL sign test, and 16 individual traits in Asian rice and 38 ones in maize that newly detected in this study deviated significantly from neutrality as well, demonstrating the dominant influence of artificial selection on them. Moreover, analysis of different categories of cross type including O. sativa × Oryza rufipogon (perennial and annual forms) crosses, maize × teosinte (Zea mays subsp. parviglumis) crosses, O. sativa × O. sativa crosses, and maize × maize crosses showed that their proportions of antagonistic QTLs were 0.1869, 0.1467, 0.2649, and 0.2618 respectively. These results revealed that selection strength of domestication is significantly stronger than that of modern genetic improvement. However, interestingly, the proportion of antagonistic QTLs (0.1591) in maize × maize with long-term selection was very similar to that (0.1467) in the maize × teosinte (Zea mays subsp. parviglumis) crosses. It suggested that some favorable traits could be cultivated within a few decades if we carry out strong selection. In addition, the proportions of antagonistic QTLs of the widely cultivated hybrids of rice (Minghui 63 × Zhenshan 97) and maize (Zheng 58 × Chang 7-2) in China were 0.309 and 0.3472 respectively. It suggested that selection during modern genetic improvement has significantly acted on them.     

Effect of Metals on Decolorization of Reactive Blue HERD by <Emphasis Type="Italic">Comamonas</Emphasis> sp. UVS

Comamonas sp. UVS was able to decolorize Reactive Blue HERD (RBHERD) dye (50 mg L^?1) within 6 h under static condition. The maximum dye concentration degraded was 1,200 mg L^?1 within 210 h. A numerical simulation with the model gives an optimal value of 35.71?±?0.696 mg dye g^?1 cell h^?1 for maximum rate (V_max) and 112.35?±?0.34 mg L^?1 for the Michaelis constant (K_m). Comamonas sp. UVS has capability of decolorization of RBHERD in the presence of Mg²⁺, Ca²⁺, Cd²⁺, and Zn²⁺, whereas decolorization was completely inhibited by Cu²⁺. Metal ions also affected the levels of biotransformation enzymes during decolorization of RBHERD. Comamonas sp. UVS was also able to decolorize textile effluent with significant reduction in COD. The biodegradation of RBHERD dye was monitored by UV–vis spectroscopy, FTIR spectroscopy, and HPLC.     

Remarks on genetic diversity and relationship of <Emphasis Type="Italic">Punica protopunica</Emphasis> and <Emphasis Type="Italic">P. granatum</Emphasis> assessed by molecular analyses

Here, two Punica species, viz., P. protopunica Balf. fil., reported as native to Socotra, and P. granatum L., were compared for the first time. Analysis of one P. protopunica and eleven P. granatum accessions was performed using three molecular markers, i.e., sequence related amplified polymorphism (SRAP), target region amplification polymorphism (TRAP), and intron targeted amplified polymorphism (ITAP), along with analysis of pgWD40 sequences, a gene involved in anthocyanin biosynthesis. All markers revealed the relationship between the two species and placed them at 33% similarity. SRAP, TRAP, and ITAP generated a total of 299, 260, and 160 bands, respectively. Of these, 78, 74, and 41 bands were specific for P. protopunica, and 92, 85, and 57 bands, respectively, were shared between both species. Sequence analysis of pgWD40~870 bp amplicons exhibited 100% identity among P. granatum accessions and 98% identity to that of P. protopunica. Phylogenetic analysis of WD40 sequences from monocot and dicot species, including both Punica species confirmed the relation between P. protopunica and P. granatum, supporting earlier reports that P. protopunica could be an ancestral species of P. granatum. Furthermore, the genetic diversity among and within P. granatum accessions from Egypt (3), Mexico (5), and Yemen (3) was assessed. Molecular marker-based relationships among region-bulked accessions was approximately the same (~90% similarity), whereas the degree of genetic variation was altered within each region. Specific bands (alleles) for accessions of each region along with those shared among them were identified. Thus, these bands could be used for pomegranate genotyping and breeding programs.     

Study Potential of Indigenous <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> and <Emphasis Type="Italic">Bacillus subtilis</Emphasis> in Bioremediation of Diesel-Contaminated Water

Petroleum products which are used in a wide variety of industries as energy sources and raw materials have become a major concern in pollution of terrestrial and marine environments. The purpose of this study was to assess the potential of indigenous microbial isolates for degradation of diesel fuel. Two most proficient bacterial strains among five isolated strains from polluted soil of an industrial refinery were studied. The isolates then were identified as Pseudomonas aeruginosa and Bacillus subtilis using biochemical tests and 16S rRNA gene sequence analyses. P. aeruginosa showed higher biodegradation efficiency than B. subtilis in shaking flask containing diesel-contaminated water. P. aeruginosa and B. subtilis degraded about 87 and 75% of total hydrocarbons, respectively, in flasks containing 2% diesel and 98% water. The biodegradation efficiency of the isolates decreased as diesel contamination increased from 2 to 5%. The isolates showed significantly higher efficiency on degradation of short-chain hydrocarbons in 20 days, i.e., by using P. aeruginosa, removal efficiency of C₁₀ hydrocarbons was near 90%, while about 69% of C₂₀₊ hydrocarbons and 47% of aromatic hydrocarbons were removed. Therefore, the isolates showed high capability in biodegradation of diesel contamination of the refinery.     

Molecular characterization of <Emphasis Type="Italic">Triticum militinae</Emphasis> derived introgression lines carrying leaf rust resistance

Triticum militinae Zhuk. et Migusch. belongs to timopheevii [Triticum timopheevii (Zhuk.) Zhuk.] group of wheats with 2n = 4x = 28 chromosomes and genome formula A^tA^tGG. Triticum militinae Zhuk. et Migusch. is known to carry resistance to fungal diseases including rusts and powdery mildew. Genes from timopheevii wheat can be incorporated into cultivated wheat by either direct hybridization or through development of amphiploids. Three T. militinae derived introgression lines (ILs) Triticum Militinae Derivative (TMD) 6-4, TMD7-5 and TMD11-5 were selected for the current study based on cytological stability. All three ILs showed resistance against wide spectrum of Indian pathotypes of leaf rust. More than 1200 SSR markers were used for genotyping of ILs and parental lines. The ILs showed variable and multiple introgressions in different chromosomes of A, B and D genome of wheat. The introgression points were distributed mostly in the distal regions though significant introgressions were also observed in proximal regions of some chromosomes. The extent of introgression in ILs TMD6-4, TMD7-5 and TMD11-5 was 2.8, 8.3 and 8.6% respectively. The set of ‘informative markers’ in the Molecularly Tagged Chromosome Regions (MTCR) of T. militinae origin can also be used in future for tagging of genes associated with traits of economic importance apart from leaf rust resistance. The transferability of Triticum aestivum L. SSR markers to T. militinae was 96.4% for A genome, 95.8% for B genome and 84.3% for D genome. Transferability of wheat SSR markers to T. militinae can be used in preparing genetic maps in timopheevii group of wheats.