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.     

Estimation of natural outcrossing rate and genetic diversity in Lima bean (<Emphasis Type="Italic">Phaseolus lunatus</Emphasis> L. var. <Emphasis Type="Italic">lunatus</Emphasis>) from Brazil using SSR markers: implications for conservation and breeding

Lima bean (Phaseolus lunatus L.) is an important food source in Brazil, especially in the northeast region, where its production and consumption are high. The goals of the present study were to estimate natural outcrossing rates and genetic diversity levels of Lima bean from Brazil, using ten microsatellite loci to obtain information for their conservation and breeding. Fourteen accessions were selected from an experiment in field with open-pollinated and with the presence of pollinating insects. Twelve seeds of each of the 14 selected accessions were grown in screenhouse for tissue harvest and DNA extraction. The multilocus model was used to determine the reproductive system. The outcrossing rate was 38.1 % (t_m = 0.381; t_s = 0.078), and the results indicated a mixed mating system with a predominance of selfing (1 ? t_m = 61.9 %). The biparental inbreeding rate was high (t _m  ? t _s  = 0.303) and the multilocus correlated paternity was quite high (r _p(m) = 0.889), indicating that the progeny was mostly composed of full sibs. The average effective number of pollen donors per maternal plant (N _ep ) was low (1.12), and the fixation index for maternal genotypes (F _m ) was 0.945, indicating that most genitors resulted from inbreeding. The studied families presented considerable genetic variability: A = 6.10;  %P = 30; H _e  = 0.60 and H _o  = 0.077. Total diversity was high (H _T = 0.596), and a portion was distributed within families (H _S = 0.058). In addition, diversity was higher between families (D _ST = 0.538), and genetic differentiation was high (G _ST = 0.902). The results presented here can be used in the implementation of Lima bean conservation and breeding programs in Brazil.     

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.     

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.     

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.     

Genetic relationships among improved varieties of rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) in Indonesia over the last 60 years as revealed by morphological traits and DNA markers

Morphological traits and two kinds of molecular markers were employed to study the genetic relationships among improved rice (Oryza sativa ) varieties of Indonesia since 1943. Dendrograms based on morphological traits and both molecular markers (simple sequence repeats, SSR and single nucleotide polymorphism, SNP) agreed in separating the varieties into two primary groups. Based on the morphological traits, a larger group (>60 %) contains varieties with smaller sizes compared with those in the smaller group (<40 %). SSR and SNP markers revealed that most of the varieties belonged to indica (88; 89 %) and japonica (9; 8 %) subspecies, and 3 % of varieties were not involved in two subspecies. The molecular markers revealed that the genetic diversity (H) stagnated between stage II (1967–1985) and stage III (1986–2003). However, during stage I (1943–1966), H was higher than in the other stages as revealed by SNP markers, while H in stage I was lower than in the other stages as revealed by SSR markers. In this study, the two molecular data sets were positively correlated and positive correlations between the phenotypic and molecular data depended on the kind of molecular marker: SNP had higher Mantel r values than SSRs. Besides, SSR markers seem to be appropriate for pedigree studies, while SNP markers could be used to reveal genomic relationships. These findings were attributable to the different properties of these two different markers. These results suggested that the diversity and differentiation of both the phenotypic and molecular marker variations were probably resulted from the crossing and selection in rice breeding in Indonesia. We suggest that Indonesia needs another strategy to improve new varieties to avoid a reduction in genetic diversity and similarity.     

Phylogenetic relationship of 40 species of genus Aloe L. and the origin of an allodiploid species revealed by nucleotide sequence variation in chloroplast intergenic space and cytogenetic insitu hybridization

Aloe species, which have been used as medicinal plants, belong to the Asphodelaceae family consisting of 530 species. In this study, genetic diversity and phylogenetic relationships among 40 Aloe species including a putative interspecies hybrid were analyzed using PCR band profiles from eight chloroplast intergenic space markers and nucleotide sequence diversity in the psbK–psbI intergenic region. A phylogenetic tree based on psbK–psbI sequences supported the revised classification of the genus Aloe as polyphyletic with several species be re-allocated into three genera Kumara, Aloidendron, and Aloiampelos. Further, the origin of the putative interspecies Aloe hybrid was characterized through molecular cytogenetics. Fluorescence and genomic insitu hybridization illustrated that the hybrid has a bimodal karyotype with a chromosome complement of 2n = 14, of which complementary halves were derived from two parental species, A. vera and A. arborescens. These findings revealed that the hybrid species was allodiploid. The phylogenetic analysis showed that A. arborescens was the maternal genome donor of the hybrid, as both have identical chloroplast genome sequences. We thus conclude that the allodiploid hybrid should be called A. arborescens × A.vera.     

Population genetics of traditional landraces of <Emphasis Type="Italic">Cucurbita pepo</Emphasis> L., 1753 in the cloud forest in Baja Verapaz,Guatemala

Little is known regarding the effect of fragmentation and human agricultural management on the genetic variation and gene flow of Cucurbita pepo L., 1753 in moderate fragmented areas in central Guatemala. We hypothesize that the genetic variation of C. pepo is affected by forest fragmentation and by traditional agricultural management. Therefore, we aim to determine: (1) the genetic diversity and genetic structure of C. pepo in the Cloud Forest Corridor (CFC) (2) the extent of genetic admixture between commercial variety (CV) and traditional landraces (TL) of C. pepo, (3) the effect of habitat fragmentation in the population genetics of C. pepo with a landscape approach, and (4) the potential relationship between traditional management practices and genetic diversity of C. pepo in the CFC. We detected the existence of high level of genetic diversity (AR = 3.43; He = 0.50), inbreeding (Fis = 0.25) and moderate population structure of C. pepo in the CFC (Fst = 0.16). No correlation between landscape and genetics was found. Also, we found high genetic admixture between CV and TL. It seems that human practices, mainly related with seed exchange patterns, could affect genetic diversity of C. pepo in the CFC. C. pepo populations in the CFC are structured, with inbreeding, and show admixture with the CV, an aspect that could affect its genetic diversity. The agricultural management influenced the population genetics of C. pepo in the CFC, but the landscape did not. We suggest that special efforts should be made to preserve the diversity of this important indigenous food source for Guatemalan people as well as their management practices.     

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.

     

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.     

Diverse morphological and cytogenetic variation and differentiation of the two subspecies in <Emphasis Type="Italic">Aegilops geniculata</Emphasis> Roth,a wild relative of wheat

Aegilops geniculata Roth, a wild relative of wheat (2n = 4x = 28, genome UUMM), is distributed over the Mediterranean basin and nearby areas. The species consists of two subspecies, subsp. geniculata and subsp. gibberosa (Zhuk.) Hammer. The former is distributed over the whole species area and has been genetically analyzed, and the latter is endemic to Spain and North Africa and has not been genetically evaluated. In this study, to clarify the genetic variation and delineation of the two subspecies from a biosystematic viewpoint, morphological variation among 23 accessions of subsp. geniculata and three of subsp. gibberosa and chromosome pairing at meiosis and fertility in their intra- and inter-subspecific F₁ hybrids were examined. A principal component analysis based on the 11 spike characteristics clearly divided the 26 accessions into two groups representing the two subspecies. The inter-subspecific F₁ hybrids showed significantly lower frequencies of chromosome pairing, significantly higher frequencies of multivalents, and significantly lower fertilities relative to those of the intra-subspecific F₁ hybrids. It was concluded that wide-ranging cytogenetic variation is included in subsp. geniculata, that subsp. gibberosa, the intra-subspecific variation of which is small, is morphologically and cytogenetically differentiated from subsp. geniculata beyond the range of the intra-subspecific variation of subsp. geniculata, and that the two subspecies are effectively isolated reproductively by hybrid sterility. The results strongly suggested that western North Africa is one of the important diversity centers of Ae. geniculata, where two subspecies were differentiated in the past and grow together in the present.     

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.     

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.     

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.     

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.     

Changes in soil microbial substrate utilization in response to altered litter diversity and precipitation in a Mediterranean shrubland

This study aimed at quantifying the consequences of reduced precipitation and plant diversity on soil microbial community functioning in a Mediterranean shrubland of southern France. Across a natural gradient of shrub species diversity, we established a total of 92 plots (4 × 4 m) with and without a moderate rain exclusion treatment of about 12 % of total precipitation. Shrub diversity included all possible combinations of the four dominant species (Cistus albidus, Quercus coccifera, Rosmarinus officinalis, and Ulex parviflorus). Respective leaf litter mixtures of these species combinations were exposed in all plots over 2 years. We quantified how litter species richness and the reduction in precipitation affected the soil microbial substrate utilization (measured by CO₂ evolution using the MicroResp method) on soil samples collected underneath each individual litter mixture after 1 and 2 years of decomposition. Moderate precipitation reduction had a minor impact, but litter species richness and the dissimilarity in phenolic concentrations (estimated using Rao’s quadratic entropy) showed a positive effect on the diversity of substrates metabolized by the microbial communities. Moreover, litter species richness increased soil microbial activity by increasing the catabolic diversity of the soil microbial community. These effects were mostly driven by the presence of Quercus and Ulex leaf litter, which at the same time reduced microbial metabolic dominance, while the presence of Rosmarinus had opposite effects. Our data suggest that plant species loss can have stronger effects on the functioning of soil microbial communities than moderate drought, with potentially important feedbacks on biogeochemical cycling in Mediterranean shrubland ecosystems.     

Nuclear and cytoplasmic contributions from <Emphasis Type="Italic">Erianthus arundinaceus</Emphasis> (Retz.) Jeswiet in a sugarcane hybrid clone confirmed through genomic in situ hybridization and cytoplasmic DNA polymorphism

A hybrid between Erianthus arundinaceus (Retz.) Jeswiet and Saccharum spontaneum L. which are wild related species of sugarcane (Saccharum L., Family Poaceae), was repeatedly crossed as female parent with sugarcane commercial varieties to develop near commercial sugarcane clones. The cytoplasm type of the hybrid derivatives were confirmed to be of E. arundinaceus through the mitochondrial and chloroplast DNA polymorphism of nad 4/3-4 intron segment and psbC–trnS segment, respectively. The E. arundinaceus × S. spontaneum hybrid with somatic chromosome number 2n = 62 was confirmed to have 30 chromosomes from E. arundinaceus through genomic in situ hybridization (GISH). The (E. arundinaceus × S. spontaneum) × sugarcane hybrid (2n = 118) had 24 chromosomes from E. arundinaceus whereas its next generation hybrid with sugarcane (2n = 108) had only 12 Erianthus chromosomes. The commercial sugarcane hybrid Co 15015, which is the third generation hybrid with 2n = 106 was confirmed to have two E. arundinaceus chromosomes through GISH. It is the first report of sugarcane with both alien cytoplasm and chromosome contributions from E. arundinaceus.     

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.     

Geographic distance and amorphous iron affect the abundance and distribution of <Emphasis Type="Italic">Geobacteraceae</Emphasis> in paddy soils in China

Purpose

Geobacteraceae are important dissimilatory Fe (III)-reducing microorganisms, influencing the cycling of metals, nutrients as well as the degradation of organic contaminants. However, little is known about their distribution, diversity, and abundance of Geobacteraceae and the effects of environment factors and geographic distance on the distribution and diversity of Geobacteraceae in paddy soils remain unclear. Therefore, the objectives of this study were to investigate the distribution, diversity, and abundance of Geobacteraceae in paddy soils and to determine key factors in shaping the Geobacteraceae distribution, environmental factors, geographic distance, or both and to quantify their contribution to Geobacteraceae variation.

Materials and methods

Illumina sequencing and quantitative real-time PCR using a primer set targeting 16S rRNA genes of bacteria affiliated with the family Geobacteraceae were employed to measure the community composition, diversity, and abundance patterns of 16S rRNA genes of Geobacteraceae in 16 samples collected from north to south of China. MRT, Mantel test, and VPA were used to analyze the relationship between communities of Geobacteraceae and environmental factors and geographic distance.

Results and discussion

Quantitative PCR showed that the abundance of 16S rRNA genes of Geobacteraceae ranged from (1.20?±?0.18)?×?10⁸ to 1.13?×?10⁹?±?2.25?×?10⁸ copies per gram of soil (dry weight) across different types of soils. Illumina sequencing results showed Geobacter was the dominant genus within the family of Geobacteraceae. Multivariate regression tree (MRT) analysis showed that soil amorphous iron contributed more (22.46 %) to the variation of dominant species of Geobacteraceae than other examined soil chemical factors such as pH (14.52 %), ammonium (5.12 %), and dissolved organic carbon (4.74 %). Additionally, more geographically distant sites harbored less similar communities. Variance partitioning analysis (VPA) showed that geographic distance contributed more to the variation of Geobacteraceae than any other factor, although the environmental factors explained more variation when combined. So, we detected the uneven distribution of Geobacteraceae in paddy soils of China and demonstrated that Geobacteraceae community composition was strongly associated with geographic distance and soil chemical factors including aFe, pH, Fe, DOC, C:N, and NO₃ ^?-N. These results greatly expand the knowledge of the distribution of Geobacteraceae in environments, particularly in terrestrial ecosystems.

Conclusions

Our results showed that geographic distance and amorphous iron played important roles in shaping Geobacteraceae community composition and revealed that both geographic distance and soil properties governed Geobacteraceae biogeography in paddy soils. Our findings will be critical in facilitating the prediction of element cycling by incorporating information on functional microbial communities into current biogeochemical models.

     

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).