Revision of Mecinus heydenii species complex (Curculionidae): integrative taxonomy reveals multiple species exhibiting host specialization

A combined taxonomic, morphological, molecular and biological study revealed that the species presently named Mecinus heydenii is actually composed of five different species: M. heydenii Wencker, 1866; M. raphaelis Baviera & Caldara sp. n., M. laeviceps Tournier, 1873; M. peterharrisi To?evski & Caldara sp. n. and M. bulgaricus Angelov, 1971. These species can be distinguished from each other by a few subtle characteristics, mainly in the shape of the rostrum and body of the penis, and the colour of the integument. The first four species live on different species of Linaria plants, respectively, L. vulgaris (L.) P. Mill., L. purpurea (L.) P. Mill. L. genistifolia (L.) P. Mill. and L. dalmatica (L.) P. Mill., whereas the host plant of M. bulgaricus is still unknown. An analysis of mtCOII gene sequence data revealed high genetic divergence among these species, with uncorrected pairwise distances of 9% between M. heydenii and M. raphaelis, 11.5% between M. laeviceps, M. heydenii and M. raphaelis, while M. laeviceps and M. peterharrisi are approximately 6.3% divergent from each other. Mecinus bulgaricus exhibits even greater divergence from all these species and is more closely related to M. dorsalis Aubé, 1850. Sampled populations of M. laeviceps form three geographical subspecies: M. laeviceps laeviceps, M. laeviceps meridionalis To?evski & Jovi? and M. laeviceps corifoliae To?evski & Jovi?. These subspecies show clear genetic clustering with uncorrected mtDNA COII divergences of approximately 1.4% from each other.     

Elucidation of the opening of the epoxidic ring of the 3beta-acetoxy-14alpha,15alpha-epoxy-5alpha-cholest-8-en-7-one by methanol, using NMR techniques assisted by a conformational study through theoretical calculations

This paper demonstrates that the crystallization of 3beta-acetoxy-14alpha,15alpha-epoxy-5alpha-cholest-8-en-7-one from methanol affords the 3beta-acetoxy-9alpha-methoxy-15alpha-hydroxycholest-8(14)-en-7-one. The structure of this steroid, which shows an apparently anomalous UV absorption maximum, is determined by high field NMR experiments, supporting the coupling constant values assignments and the NOE contacts by a conformational study through theoretical calculations at the B3LYP/6-31G* level. The computational study also justifies the observed UV absorption of the steroid, thus demonstrating the usefulness of computer chemistry in providing support for the identification of unknown compounds.     

Selective Cytochrome c Displacement by Phosphate and Ca2+ in Brain Mitochondria

In brain mitochondria, phosphate- and Ca²⁺-dependent cytocrome c (cyt c) release reveals pools that interact differently with the inner membrane. Detachment of the phosphate-dependent pool did not influence the pool released by Ca²⁺. Cyt c pools were also detected in a system of cyt c reconstituted in cardiolipin (CL) liposomes. Gradual binding of cyt c (1 nmol) to CL/2–[12-(7-nitrobenz- 2-oxa-1,3-diazol-4-yl)amino]dodecanoyl-1-hexadecan oyl-sn-glycero-3-phosphocholine (NBDC₁₂-HPC) liposomes (10 nmol) produced NBD fluorescence quenching up to 0.4 nmol of added protein. Additional bound cyt c did not produce quenching, suggesting that cyt c-CL interactions originate distinct cyt c pools. Cyt c was removed from CL/NBDC₁₂-HPC liposomes by either phosphate or Ca²⁺, but only Ca²⁺ produced fluorescence dequenching and leakage of encapsulated 8-aminonaphthalene-1,3,6-trisulfonic acid/p-xylene-bis-pyridinium bromide. In mitochondria, complex IV activity and mitochondrial membrane potential (Δψ_m) were not affected by the release of the phosphate-dependent cyt c pool. Conversely, removal of cyt c by Ca²⁺ caused inhibition of complex IV activity and impairment of Δψ_m. In a reconstituted system of mitochondria, nuclei and supernatant, cyt c detached from the inner membrane was released outside mitochondria and triggered events leading to DNA fragmentation. These events were prevented by enriching mitochondria with exogenous CL or by sequestering released cyt c with anti-cyt c antibody.     

Drug treatment in the development of mismatch repair defective acute leukemia and myelodysplastic syndrome

DNA from therapy-related acute leukemia/myelodysplastic syndrome cases (tAL/MDS) from the GIMEMA [Gruppo Italiano Malattie Ematologiche Maligne dell'Adulto] Archive was examined for the microsatellite instability (MSI(+)) phenotype that is diagnostic for defective DNA mismatch repair. More than 60% (16/25) of tAL/MDS cases were MSI(+) in contrast to <4% (0/28) of de novo cases. hMLH1 gene silencing was rare and evidence of promoter methylation was found in less than one-third of the MSI(+) cases. Among the GIMEMA patients who had been treated for breast cancer there was an apparent trend towards early onset primary breast disease. This suggests that there might be common predisposing factors for breast cancer and tAL/MDS. There were also three examples of mutations in the MRE11 gene among the 25 tAL/MDS cases suggesting that defective recombinational DNA repair may promote the development of secondary malignancy. MSI(+) tAL/MDS was significantly associated with previous chemotherapy and the frequency of MSI(+) among radiotherapy patients was considerably lower. In view of the established relationship between drug resistance and mismatch repair defects, we suggest that selection for therapeutic drug resistance may contribute to the incidence of MSI(+) tAL/MDS.     

DNA barcoding as a complementary tool for conservation and valorisation of forest resources

Since the pre-historic era, humans have been using forests as a food, drugs and handcraft reservoir. Today, the use of botanical raw material to produce pharmaceuticals, herbal remedies, teas, spirits, cosmetics, sweets, dietary supplements, special industrial compounds and crude materials constitute an important global resource in terms of healthcare and economy. In recent years, DNA barcoding has been suggested as a useful molecular technique to complement traditional taxonomic expertise for fast species identification and biodiversity inventories. In this study, in situ application of DNA barcodes was tested on a selected group of forest tree species with the aim of contributing to the identification, conservation and trade control of these valuable plant resources.The “core barcode” for land plants (rbcL, matK, and trnH-psbA) was tested on 68 tree specimens (24 taxa). Universality of the method, ease of data retrieval and correct species assignment using sequence character states, presence of DNA barcoding gaps and GenBank discrimination assessment were evaluated. The markers showed different prospects of reliable applicability. RbcL and trnH-psbA displayed 100% amplification and sequencing success, while matK did not amplify in some plant groups. The majority of species had a single haplotype. The trnH-psbA region showed the highest genetic variability, but in most cases the high intraspecific sequence divergence revealed the absence of a clear DNA barcoding gap. We also faced an important limitation because the taxonomic coverage of the public reference database is incomplete. Overall, species identification success was 66.7%.This work illustrates current limitations in the applicability of DNA barcoding to taxonomic forest surveys. These difficulties urge for an improvement of technical protocols and an increase of the number of sequences and taxa in public databases.     

Risposte Fotomorfogeniche Indotte Dalla Radiazione UV-B in Brassica Oleracea var. Capitata

Abstract

Photomorphogenic responses induced by UV-B radiation in Brassica oleracea var. capitata.—Ultraviolet radiation can induce a plethora of “damaging” and “non damaging” effects in higher plants. We analyzed two possible photomorphogenic responses to UV-B radiation, the anthocyanin accumulation and the inhibition of hypocotyl elongation by modifying the UV spectral range with specific cut-off filters, under two levels of photon fluence rate. Experimental results suggest that detrimental effects are due to shorter wavelenghts of UV-B region (less than 305 nm); in contrast some adaptative responses may be induced by longer wavelenghts of UV-B region (between 305 and 320 nm). We attempted to determine the involvment of endogenous anthocyanin content in the UV-B photoprotection. Our experiments suggest a secondary role of anthocyanin accumulation in UV-B plant adaptation.