Isolation and characterization of microsatellite markers in Acca sellowiana (Berg) Burret

Acca sellowiana has commercial potential because of the quality and the unique flavor of its fruit. Conservation of natural populations and management of breeding programmes would benefit from the availability of molecular markers that could be used to characterize levels and distribution of genetic variability. Thus, 13 microsatellite markers were developed from an enriched genomic library of A. sellowiana. They were characterized using 40 samples. The expected and observed heterozygosities ranged from 0.513 to 0.913 and from 0.200 to 0.889, respectively. These are the first microsatellite loci characterized from A. sellowiana that will contribute to improve researches on the genetic conservation, characterization and breeding.     

Three new flavonoids,proanthocyanidin, and accompanying phenolic constituents from Feijoa sellowiana

Our investigation of phenolic constituents of fruits, flower buds, and leaves of Feijoa sellowiana led to the isolation of twenty-one phenolics including three new gossypetin glycosides 1–3, and also the purification of a proanthocyanidin fraction. A high-performance liquid chromatography method for simultaneous analysis of phenolic constituents was established and then used to investigate the phenolic profiles of the parts of the plant species, to show the presence of characteristic flavonoids and ellagic acid derivatives or ellagitannins in the extracts from fruits, flower buds, and leaves. The branch extract profile also suggested the presence of alkylated ellagic acids as characteristic constituents. Inhibitory effects of feijoa flavonoids on mushroom tyrosinase were seen, although in some cases this may have resulted from direct interaction with the enzyme. Cytotoxic effect of the proanthocyanidin fraction was also shown.     

Secondary metabolites from the leaves of Feijoa sellowiana Berg

The investigation of the lipid extract of leaves of Feijoa sellowiana cultivated along the east coast of Sicily has yielded in addition to the widespread secondary metabolites: alpha-tocopherol, flavone, stigmasterol and beta-carotene, an inseparable mixture of tyrosol esters of lignoceric (1a), cerotic (1b) and montanic (1c) acids, and a novel galactolipid identified as (2S)-1,2,6'-tri-O-[(9Z,12Z,15Z)-octadeca-9,12,15-trienoyl]-3-O-beta-D-galactopyranosyl glycerol (2).     

Phytochemical profile,enzyme inhibition activity and molecular docking analysis of Feijoa sellowiana O. Berg

Feijoa sellowiana leaves and fruits have been investigated as a source of diverse bioactive metabolites. Extract and eight metabolites isolated from F. sellowiana leaves were evaluated for their enzymatic inhibitory activity against α-glucosidase, amylase, tyrosinase, acetylcholinestrerase and butyrylcholinesterase both in vitro and in silico. Feijoa leaves’ extract showed strong antioxidant activity and variable levels of inhibitions against target enzymes with a strong anti-tyrosinase activity (115.85 mg Kojic acid equivalent/g). Additionally, α-tocopherol emerged as a potent inhibitor of AChE and BChE (5.40 & 10.38 mmol galantamine equivalent/g, respectively). Which was further investigated through molecular docking and found to develop key enzymatic interactions in AChE and BChE active sites. Also, primetin showed good anti BChE (11.70 mmol galantamine equivalent/g) and anti-tyrosinase inhibition (90.06 mmol Kojic acid equivalent/g) which was also investigated by molecular docking studies.

Highlights

• Isolation of eight bioactive constituents from Feijoa sellowiana leaves.
• In vitro assays using different enzymatic drug targets were investigated.
• In silico study was performed to define compound interactions with target proteins.
• Feijoa leaf is an excellent source of anti-AChE and antityrosinase bioactives.

     

Effect and analysis of phenolic compounds during somatic embryogenesis induction in Feijoa sellowiana Berg

Summary. The effect of phenolic compounds on somatic embryogenesis in Feijoa sellowiana was analysed. The results showed that caffeic acid (140–560 μM) significantly increased somatic embryogenesis induction compared with the control. The presence of phloridzin, even at lower concentrations (11.5 μM), or caffeic acid or phloroglucinol at concentrations greater than 140.0 and 197.5 μM, respectively, inhibited somatic embryo development beyond the globular stage. When somatic embryos were transferred to the germination medium, the highest rates of germination (81.9%) were obtained with embryos induced in the presence of phloroglucinol (79.0 μM). At all concentrations tested, somatic embryos induced in medium containing phloroglucinol germinated at higher rates than those induced in the presence of caffeic acid. Histological and ultrastructural studies showed that somatic embryos were formed in close association with phenolic-rich cells which, in more advanced stages of development, formed a zone isolating the embryo from the maternal tissue. A comparative analysis of total phenolic content indicated that phenolics reached a peak by the third week of culture, independently of the medium used. However, after that period, the amount of phenolic compounds was significantly higher in explants cultured in the presence of phloroglucinol than in those cultured in the control or in caffeic acid-containing medium. Attempts to identify the type of phenolic compounds showed that flavan-3-ols and gallic acid derivatives were mainly produced in phloroglucinol-containing medium, whereas flavanones and dihydroflavonols were also present in medium containing caffeic acid. Flavones were the main phenols detected in the control. The ways in which phenolic compounds may affect somatic embryogenesis are discussed. Correspondence: J. M. Canhoto, Departamento de Botanica, Faculdade de Ciências e Tecnologia, Universidade de Coimbra, Cal?ada Martim de Freitas, 3001-455 Coimbra, Portugal.     

Isolation and characterization of microsatellite markers in Fraser fir (Abies fraseri)

We describe the isolation and characterization of 14 microsatellite loci from Fraser fir (Abies fraseri). These markers originated from cloned inserts enriched for DNA sequences containing tandem di‐ and tri‐nucleotide repeats. In total, 36 clones were selected, sequenced and evaluated. Polymerase chain reaction (PCR) primers for 14 of these sequences consistently produced simple PCR profiles and were found to be polymorphic among 13 Fraser fir samples. In addition, more than half of these loci were found to amplify a wide range of samples from several Abies taxa.     

Isolation and characterization of microsatellite markers for Araucaria angustifolia (Araucariaceae)

Araucaria angustifolia is a dioecious tree species that occurs in the southern part of Brazil. Because of the intense exploitation of the species due to its valuable wood, only 2% of the original population still remains. Twenty‐nine species‐specific and highly polymorphic microsatellite loci were developed from a genomic library enriched for AG/TC repeats. Levels of polymorphism were evaluated using a total of 16 adult trees from a natural population. An average of 8.1 alleles per locus was detected, and expected heterozygosity ranged from 0.63 to 0.72.     

Isolation and characterization of new microsatellite markers in mango (Mangifera indica)

Six microsatellite loci that were isolated from a microsatellite‐enriched genomic library of mango (Mangifera indica) along with their specific primer sets were each characterized by using 36 cultivars collected mainly in Thailand. The observed and expected heterozygosities ranged from 0 to 0.83 and from 0.29 to 0.73, respectively. The number of putative alleles are two to six. Three of the six alleles have frequencies of over 75%. The high frequency may be attributed to the bias in the origin of cultivars. Among 36 mango cultivars tested, 29 cultivars showed a unique pattern by six primer sets, whereas seven cultivars cannot be identified because of genotype similarities. This suggests the potentials for identification of mango cultivars by microsatellite markers.     

Isolation and characterization of polymorphic microsatellite markers from Primula nutans (Primulaceae)

Seven polymorphic microsatellite loci were developed for a perennial seashore plant, Primula nutans. Degenerate oligonucleotide‐primed (DOP)–polymerase chain reaction (PCR)‐amplified DNA was ligated to TOPO TA vector and screened with radioactively labelled dinucleotide repeat probes. A sample of 378 individuals from Finland, Norway and Russia were used to characterize those loci, which exhibited two to four alleles per locus with observed heterozygosity of 0.003–0.229 and expected heterozygosity of 0.016–0.527. No linkage disequilibrium was found between these seven loci. These are the first microsatellite markers reported for P. nutans.     

Isolation and characterization of microsatellite markers in tree lizards (Urosaurus ornatus)

Tree lizards (Urosaurus ornatus) are a highly accessible species useful in testing theories of life-history evolution and behavioural ecology. A polymerase chain reaction-based method was used to isolate nine polymorphic tetranucleotide microsatellite loci from the genome of these lizards. The isolated loci displayed 5 to 9 alleles in the populations screened. Observed heterozygosity ranged from 0.333 to 0.900. Results indicate these loci now provide a basis to study mate choice, parentage, and population genetic structure within this species.     

Isolation and characterization of microsatellite markers in the liver fluke (Fasciola hepatica)

Six microsatellite markers were isolated from Fasciola hepatica, a re‐emerging parasite that causes important veterinary and public health problems. In a sample of 52 liver flukes from a region of hyperendemicity (Bolivian Altiplano), five microsatellite were polymorphic. Our results showed that liver flukes present important genetic variability, suggesting a preferential outcrossing reproduction mode for this hermaphroditic parasite.     

Isolation and characterization of microsatellite markers in Bonelli's eagle (Hieraaetus fasciatus)

To obtain polymorphic molecular markers for population genetics and conservation studies in Bonelli's eagle populations, we screened a partial genomic library enriched for microsatellites with di‐ and tri‐nucleotide motifs [(GT) (CT) (AAC) and (GCC)]. A total of 15 polymorphic markers were obtained. The number of alleles ranged from two to eight. These markers will be very useful for paternity tests and population structure studies as well as for evaluating the outcome of conservation programs.     

Isolation and characterization of microsatellite markers in black muntjac (Muntiacus crinifrons)

The black muntjac (Muntiacus crinifrons) is a vulnerable species found in the mountains of eastern China, about which little is known. Here we develop 11 polymorphic microsatellite loci from a (CA)(n) -enrichment library for the animal. In the analyses of 25 individuals sampled, unbiased expected heterozygosity levels varied from 0.686 to 0.876 and the number of alleles per locus varied from five to nine. Results that 11 microsatellite loci were highly polymorphic indicated that these markers are sufficiently powerful to address such questions as parentage, mating system and population genetic structure of M. crinifrons.     

Isolation and characterization of microsatellite markers in Elliot's pheasant (Syrmaticus ellioti)

We isolated eight microsatellite loci from genomic DNA in Elliot's pheasant Syrmaticus ellioti using streptavidin‐coated magnetic beads. In the analyses of 53 individuals sampled, these loci displayed polymorphism varying from six to nine alleles per locus and observed heterozygosities ranging from 0.174 to 0.430. The results suggested that these novel microsatellite markers could become useful molecular tools for genetic studies of S. ellioti.     

Isolation and characterization of microsatellite markers in hoary marmots (Marmota caligata)

Microsatellite loci were developed from hoary marmots (Marmota caligata) to aid in the investigation of the social structure and mating system of this species. Seven of the microsatellite loci developed were found to be moderately polymorphic with between two and seven alleles per locus. In addition to the microsatellites developed in hoary marmots we also tested markers developed for other scuirids, namely European alpine marmots (M. marmota), Columbian ground squirrels (Spermophilus columbianus) and European ground squirrels (S. citellus). Of these markers, 13 were polymorphic when amplified in hoary marmots with between two and nine alleles per locus.     

Induction of pollen callus in anther cultures of Feijoa sellowiana Berg. (Myrtaceae)

Summary Anthers of Feijoa sellowiana Berg. (feijoa) produced pollen callus when cultured in Murashige and Skoog medium containing 2,4-dichlorophenoxyacetic acid and benzyladenine or in nurse cultures. Somatic callus was also formed in large amounts from the connective and from the cut end of the filament. Anthers containing microspores at the stage immediately prior to the first pollen mitosis cultured in the presence of 3% sucrose, presented the highest frequencies of induction. Androgenetic divisions were initiated by the formation of two morphologically equal cells, the so-called B-pathway. Attempts to regenerate pollen plants were unsuccessful but leaf-like structures could be obtained in regeneration media containing combinations of gibberellic acid and benzyladenine.Abbreviations 2,4-D 2,4-diclorophenoxyacetic acid - BA benzyladenine - FDA fluorescein diacetate - GA₃ gibberellic acid - Kn kinetin - MS Murashige and Skoog (1962) medium     

Isolation and characterization of 11 microsatellite markers from Sagittaria latifolia (Alismataceae)

We developed 11 microsatellite loci for Sagittaria latifolia, an aquatic plant common to wetlands of North America. From an (AG)-enriched library, we identified 66 unique microsatellite sequences for which primers could be designed. Twenty-two loci reliably amplified a clear single band of expected size, and 11 loci were scoreable and polymorphic. For these 11 loci, we genotyped a monoecious and a dioecious population, yielding four to 14 alleles per locus. Three loci exhibited significant linkage disequilibrium leaving eight independent variable loci. Eight loci also amplified in four other Sagittaria species. These microsatellite loci will be useful to compare genetic structure among monoecious and dioecious populations of S. latifolia.     

Isolation and characterization of microsatellite markers in the caddisfly Drusus discolor (Trichoptera: Limnephilidae)

We describe the development of and amplification conditions for microsatellite primers isolated from the caddisfly Drusus discolor. Eight polymorphic microsatellite loci were developed and screened for variability using 37 individuals from two populations from central Europe. The primers yielded an average of 8.6 alleles per loci. No linkage disequilibrium between loci was detected, while three loci showed deviations from Hardy–Weinberg equilibrium in one of the two tested populations.