<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.     

Microbial and Physico-chemical Characteristics Associated with the Incidence of <Emphasis Type="Italic">Legionella</Emphasis> spp. and <Emphasis Type="Italic">Acanthamoeba</Emphasis> spp. in Rainwater Harvested from Different Roofing Materials

The incidence of Legionella and Acanthamoeba spp. was correlated to microbial indicator analysis and physico-chemical characteristics of rainwater harvested from catchment areas constructed from galvanized zinc, Chromadek®, and asbestos, respectively. Quantitative PCR (qPCR) analysis indicated that no significant difference (p?>?0.05) in copy numbers of Legionella spp. and Acanthamoeba spp. was recorded in tank water samples collected from the respective roofing materials. However, significant positive Spearman (ρ) correlations were recorded between the occurrences of Legionella spp. gene copies vs. nitrites and nitrates (p?=?0.05) in all tank water samples. Significant positive correlations were also established between Acanthamoeba spp. vs. barium (p?=?0.03), magnesium (p?=?0.02), sodium (p?=?0.02), silicon (p?=?0.05), arsenic (p?=?0.03), and phosphate (p?=?0.01), respectively. Additionally, while no significant correlations were observed between Legionella spp. vs. the indicator bacteria (p?>?0.05), positive correlations were observed between Acanthamoeba spp. vs. total coliforms (p?=?0.01) and Acanthamoeba spp. vs. Escherichia coli (p?=?0.02), respectively. Results obtained in the current study thus indicate that the incidence of Acanthamoeba and Legionella spp. in harvested rainwater was not influenced by the roofing material utilized. Moreover, it is essential that the microbial quality of rainwater be assessed before this water source is implemented for potable and domestic uses as untreated harvested rainwater may lead to legionellosis and amoebae infections.     

Polymorphism of <Emphasis Type="Italic">Got2</Emphasis> DNA sequences sheds light on <Emphasis Type="Italic">Aegilops tauschii</Emphasis> Coss. intraspecies divergence and origin of <Emphasis Type="Italic">Triticum aestivum</Emphasis> L.

DNA sequences of nuclear gene Got2 was studied in 60 accessions of Aegilops tauschii, 29 of subsp. tauschii and 31 of subsp. strangulata. It was found that Got2 allozyme polymorphism in Ae. tauschii is due to a single, unique, mutation which led to replacement of glutamic acid by isoleucine in residue 256 of the enzyme molecule, encoded by Got2. As revealed by Got2 DNA sequences variation, initially in its history Ae. tauschii was presented by subsp. strangulata, and among phylogenetic lineages of subsp. strangulata, the lineage “t-9¹s” (TauL3) is the most ancient, a relict one. Subspecies tauschii is relatively “young”. Initially it was presented by the lineage marked by combination of allozyme alleles Got2 ¹⁰⁵ and Acph1 ¹⁰⁰. In the past it inhabited the Continental area from Caucasia to Pakistan, but later on it was forced out by newly originated, now—a major lineage of subsp. tauschii, marked by Got2 ¹⁰⁰. This lineage extended the Continental area of the species up to Kirgizstan, but actually failed to penetrate into pre-Caspian area, occupied by subsp. strangulata. These results essentially differ from those obtained previously, using chloroplast DNA (cpDNA) sequences polymorphism. As revealed by cpDNA, the major, “usual”, subsp. strangulata (TauL2) is “younger” than subsp. tauschii, which resided on phylogenetic tree between relict lineage “t-9¹s”of subsp. strangulata—and major subsp. strangulata. But both cpDNA and Got2 DNA sequences indicate that the level of genetic variation in subsp. tauschii is much lower than in subsp. strangulata. According to Got2 DNA sequences variation, it was Ae. tauschii subsp. strangulata lineage “k-109″ which donated genome D to Triticum aestivum L. This lineage includes accessions: k-109 from South-Eastern Precaspian Azerbaijan; KU-2105, KU-2159 from Western Precaspian Iran; KU-2080 from Eastern Precaspian Iran.     

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.     

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.     

Multitrophic Assessment of Copper Contamination Using Watercress (<Emphasis Type="Italic">Nasturtium officinale</Emphasis>) and the Mustard Beetle (<Emphasis Type="Italic">Phaedon viridis</Emphasis><Emphasis Type="Bold">)</Emphasis>

Copper is an active component of some commercial algaecides and is commonly found in low concentrations in contaminated aquatic systems. Unintended consequences of algaecide application include macrophyte bioaccumulation and possible trophic level bioamplification especially by specialist herbivores. Trophic level effects of copper contamination were observed through feeding trials using the mustard beetle (Phaedon viridis). Several metals, including copper, interfere with the myrosinase enzyme system responsible for the watercress (Nasturtium officinale) allelopathic defense against herbivory. The mustard beetle is a specialist herbivore that has evolved a detection system that is stimulated by the products of the glucosinolate-myrosinase system. Because copper interferes with myrosinase enzymes, mustard beetles were expected to avoid copper-contaminated plants. While larvae exhibited a slight preference for contaminated plants, adult mustard beetles in this experiment exhibited a statistically significant preference for plants uncontaminated with copper.     

Effects of ectomycorrhizal fungus <Emphasis Type="Italic">Boletus edulis</Emphasis> and mycorrhiza helper <Emphasis Type="Italic">Bacillus cereus</Emphasis> on the growth and nutrient uptake by <Emphasis Type="Italic">Pinus thunbergii</Emphasis>

The present greenhouse study was undertaken to evaluate the effects of co-inoculating the ectomycorrhizal (ECM) fungus Boletus edulis with the mycorrhiza helper bacterium Bacillus cereus HB12 or HB59 on the growth and nutrient uptake of Pinus thunbergii. The inoculation with mycorrhiza helper bacterium significantly (P?≤?0.05) increased the ectomycorrhizal colonization. Treatments with dual inoculum (the mycorrhiza helper bacterium plus mycorrhiza) significantly (P?≤?0.05) increased the P. thunbergii growth. Bacteria–mycorrhizae interactions resulted in a great utilization of phosphate and potassium. The single inoculation resulted in a higher root activity than the control while the co-inoculation led to the highest root activity. The 6-CFDA staining assay showed that B. cereus enhanced fungal activity in ectomycorrhizal symbiosis. The results conclusively suggest that B. cereus isolated from the rhizosphere of P. thunbergii can potentially be used as individual inoculant or co-inoculated with ECM fungi to increase the production in sustainable ecological systems. These results support the potential use of B. cereus (HB12 or HB59) and B. edulis as mixed inoculants stimulating growth of P. thunbergii.     

Identification of new sources of resistance to <Emphasis Type="Italic">Striga gesnerioides</Emphasis> in cowpea <Emphasis Type="Italic">Vigna unguiculata</Emphasis> accessions

The parasitic weed, Striga gesnerioides, is a major threat to cowpea productivity throughout the savannas of West and Central Africa. The identification of sources of S. gesnerioides resistance and their incorporation into breeding programs would be a beneficial strategy to combat the devastation caused by the parasite in cowpea fields. In this study we examined one hundred and ninety-four (194) accessions, four commercial varieties and two controls collected from a mini core collection of cowpea held at the International Institute of Tropical Agriculture genebank for resistance to S. gesnerioides race 3 (SG3), the most prevalent race in Nigeria, using phenotypic screening and molecular marker analysis. Our studies identified two cowpea accessions, Tvu-1272 and Tvu-16514, that are resistant to S. gesnerioides SG3. Resistance in these lines is associated with the molecular marker SSR1, known to segregate with the gene conferring resistance to SG3 in the cultivar B301. Phenotypically, resistance in Tvu-1272 and Tvu-16514 is expressed as a hypersensitive response at the site of infection on the roots. Allelism tests indicated that the gene that conferring SG3 resistance in Tvu-1272 is independent of that conferring resistance in B301. Tvu-1272 and Tvu-16514 will provide additional new sources of resistance to Striga and races prevalent in Nigeria.     

Diversity of <Emphasis Type="Italic">Citrus</Emphasis><Emphasis Type="Italic">depressa</Emphasis> Hayata (Shiikuwasha) revealed by DNA analysis

Citrus depressa Hayata is an indigenous mandarin species on the Ryukyu Islands located in the subtropical region of Japan. We deduced its phylogenetic relationships by evaluating accessions grown on various Ryukyu Islands via cleaved amplified polymorphic sequence analysis of cpDNA and sequence-related amplified polymorphism (SRAP). The cpDNA results indicated that C. depressa could be classified into two types. SRAP revealed patterns of diversity within C. depressa consistent with our cpDNA results. These results indicate that maternal origin may influence or is correlated with the constitution of the nuclear genome of C. depressa. Another Japanese mandarin species, Citrus tachibana (Makino) Tanaka was distinguished from C. depressa by SRAP markers. Moreover, both C. depressa and C. tachibana could be distinguished from other Citrus species. Our results suggest that Japanese mandarin possesses a characteristic genome with the genus Citrus.     

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.     

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.     

Genetic differentiation,races and interracial admixture in avocado (<Emphasis Type="Italic">Persea americana</Emphasis> Mill.), and <Emphasis Type="Italic">Persea</Emphasis> spp. evaluated using SSR markers

Avocado (Persea americana Mill.) is a subtropical domesticated fruit tree indigenous to Mesoamerica. It is a member of the Lauraceae family and is separated into three horticultural races (Guatemalan, Mexican, and West Indian) mainly corresponding to their ecological adaptation, botanical, and physiological traits. Main objectives of this study were to characterize the population structure, genetic diversity, and horticultural race of a total of 354 Persea spp. trees whose origin is as follow: 221 trees [P. americana, (218), P. nubigena (2) and P. krugii (1)] from the USDA-ARS-Subtropical Horticultural Research Station, Miami; 105 trees from the Fairchild Farm [P. americana (104) and P. schiedeana (1)], and 28 trees collected in Mexico [P. schiedeana (23) and P. americana (5)]. The complexity of their interracial admixture; as well as mislabeling frequency was also evaluated. Molecular marker analysis utilizing a set of 55 simple sequence repeat (SSR) markers amplified a total of 869 alleles with a mean number of alleles per locus of 15.8 and average polymorphism information content value of 0.71, indicating a high variability in the allele frequency for the collection. Significant deviations from Hardy–Weinberg equilibrium were identified after Bonferroni correction for a large number of loci (48; 87%) due to the presence of null alleles. The main source of variation for this population was found to be within individuals (66.84%), with 19.30% variation among populations, and 13.86% variation among individuals within populations. Moreover, population specific inbreeding indices (F _IS ) were calculated for West Indian, Guatemalan, and Mexican [(0.1918; p value 0.0000), (0.1879; p-value 0.0000), (0.0925; p-value 0.0022)], respectively. Bayesian analysis divided the individual genotypes into groups associated with the Guatemalan, Mexican, West Indian races; interracial admixture; complex hybrids and P. schiedeana species. Also, results of the multivariate clustering method (PCA) and genetic distance analyses calculated among all possible individual combinations within the SSR diversity data agreed with Bayesian or Structure analyses results. The 55 SSRs provided complete resolution of all individuals and the estimated mislabeling error was approximately 0.28%.     

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.     

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).

     

Combined bioremediation of soil co-contaminated with cadmium and endosulfan by <Emphasis Type="Italic">Pleurotus eryngii</Emphasis> and <Emphasis Type="Italic">Coprinus comatus</Emphasis>

Purpose

The subjects of this study were to investigate the remediating potential of the co-cultivation of Pleurotus eryngii and Coprinus comatus on soil that is co-contaminated with heavy metal (cadmium (Cd)) and organic pollutant (endosulfan), and the effects of the co-cultivated mushrooms on soil biochemical indicators, such as laccase enzyme activity and bacterial counts.

Materials and methods

A pot experiment was conducted to investigate the combined bioremediation effects on co-contaminated soil. After the mature fruiting bodies were harvested from each pot, the biomass of mushrooms was recorded. In addition, bacterial counts and laccase enzyme activity in soil were determined. The content of Cd in mushrooms and soil was detected by the flame atomic absorption spectrometry (FAAS), and the variations of Cd fractions in soil were determined following the modified BCR sequential extraction procedure. Besides, the residual endosulfan in soil was detected by gas chromatography-mass spectrometry (GC-MS).

Results and discussion

The results indicated that co-cultivation of P. eryngii and C. comatus exerted the best remediation effect on the co-contaminated soil. The biomass of mushroom in the co-cultivated group (T group) was 1.57–13.20 and 19.75–56.64% higher than the group individually cultivated with P. eryngii (P group) or C. comatus (C group), respectively. The concentrations of Cd in the fruiting bodies of mushrooms were 1.83–3.06, 1.04–2.28, and 0.67–2.60 mg/kg in T, P, and C groups, respectively. Besides, the removal rates of endosulfan in all treatments exceeded 87%. The best bioremediation effect in T group might be caused by the mutual promotion of these two kinds of mushrooms.

Conclusions

The biomass of mushroom, laccase activity, bacterial counts, and Cd content in mushrooms were significantly enhanced, and the dissipation effect of endosulfan was slightly higher in the co-cultivated group than in the individually cultivated groups. In this study, the effect of co-cultivated macro fungi P. eryngii and C. comatus on the remediation of Cd and endosulfan co-contaminated soil was firstly reported, and the results are important for a better understanding of the co-remediation for co-contaminated soil.

     

The genetic relationship and structure of some natural interspecific hybrids in <Emphasis Type="Italic">Prunus</Emphasis> subgenus <Emphasis Type="Italic">Prunophora</Emphasis>, based on nuclear and chloroplast simple sequence repeats

In the subgenus Prunophora of the genus Prunus, many transitional traits presented in interspecific hybrids, the so-called ‘new species’, were frequently named due to the complicated botanical classification system. In this study, we used 16 nuclear simple sequence repeats (nSSRs) and 10 chloroplast simple sequence repeats (cpSSRs) to evaluate genetic relationships among 42 accessions, which included 15 putative interspecific hybrids, and then to reveal the speciation and differentiation in the subgenus Prunophora. In total, 231 and 27 alleles were observed in nSSRs and cpSSRs respectively; and with cpSSRs 20 haplotypes were revealed among the accessions. Furthermore, the haplotype and genetic structure analysis implied that (1) Prunus simonii Carr. might be a subspecies or a forma of Prunus salicina L., rather than an interspecific hybrid between P. salicina and Prunus armeniaca L., (2) Prunus limeixing Zhang et Wang was derived from a natural hybrid with P. salicina as its maternal progenitor and P. armeniaca as the female parent, and (3) Prunus cathayana Fu et al. (or kernel-using apricot) was an interspecific hybridization species of P. armeniaca (maternal parent) and Prunus sibirica L. (female parent). These results will be useful for clarifying the problems in the botanical classification, and facilitate the conservation and management of plum and apricot genetic resources in the Chinese National Germplasm Repository for Plums and Apricots.     

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.     

Agro-morphological traits of <Emphasis Type="Italic">Cicer reticulatum</Emphasis> Ladizinsky in comparison to <Emphasis Type="Italic">C. echinospermum</Emphasis> P.H. Davis in terms of potential to improve cultivated chickpea (<Emphasis Type="Italic">C. arietinum</Emphasis> L.)

Germplasm collections of C. reticulatum and C. echinospermum are limited, while both species face threats from over-grazing and habitat change in their natural environments. Recently many new accessions of C. reticulatum and C. echinospermum were collected in east and south-east Anatolia (Turkey) but they have not yet been evaluated for agro-morphological traits. Therefore, the current study investigated agro-morphological traits of new germplasm sources of C. reticulatum and C. echinospermum and evaluated resistance to biotic and abiotic stresses for chickpea improvement. The most attractive agro-morphological traits were canopy width, number of stems and pods per plant and biological yield. The most productive accessions of C. reticulatum and C. echinospermum had 712 and 625 pods per plant, respectively. Two distinct seed, flower and leaf shapes were found in accessions of C. echinospermum. Path analyses indicated that biological yield and harvest index had the most direct influence on seed yield in both species. Factor analyses showed that high seed yield in C. reticulatum depended on high biological yield and number of pods per plant, whereas high seed yield in C. echinospermum depended on harvest index. It was concluded that most accessions of C. reticulatum and C. echinospermum were not only resistant to some biotic and abiotic stresses but also had hidden alleles that could produce transgressive segregation in crosses to cultivated material.