Anna Scialabba

Effects of ageing on peroxidase activity and localization in radish ( Raphanus sativus L.) seeds

Peroxidase activity was assayed in crude extracts of integument, cotyledons and embryo axis of radish seeds, deteriorated under accelerated ageing conditions. Over five days of ageing, in which germination decreased from 100 to 52%, the enzyme activity in integument was higher than that in other seed parts, increasing in the first days of ageing and then decreasing sharply in extremely aged seeds. Polyacrylamide gel electrophoresis analysis showed four peroxidase isoenzymes with MM of 98, 52.5, 32.8 and 29.5 kDa in the embryo axis of unaged seeds, and only the 32.8 and 29.5 kDa MM isoforms in the integument and cotyledons. In these parts of the seed, only the 29.5 kDa MM isoenzyme increased in activity in early days of ageing and decreased there-after. In the embryo axis, the 29.5 kDa MM isoenzyme activity increased slowly in the first day of ageing, while the 98 and 52.5 kDa MM isoenzyme activities disappeared. A cytochemical localization of peroxidase activity in the various tissues showed that main differences between unaged and extremely aged seeds occurred in the embryo axis.

Ecotoxicity of halloysite nanotube–supported palladium nanoparticles in Raphanus sativus L

Halloysite nanotubes (HNTs) are natural nanomaterials that are biocompatible and available in large amounts at low prices. They are emerging nanomaterials with appealing properties for applications like support for metal nanoparticles (NPs). The potential environmental impacts of NPs can be understood in terms of phytotoxicity. Current research has been focusing on HNT applications in cell or animal models, while their use in plants is limited so their ecotoxicological impact is poorly documented. To date there are no studies on the phytotoxic effects of functionalized halloysites (functionalized-HNTs). To develop a quantitative risk assessment model for predicting the potential impact of HNT-supported palladium nanoparticles (HNT-PdNPs) on plant life, an investigation was undertaken to explore their effects on seed germination, seedling development, and mitotic division in root tip cells of 2 lots of Raphanus sativus L. with different vigor. The results showed that exposure to 1500 mg/L of HNTs, functionalized-HNTs, and HNT-PdNPs had no significant influence on germination, seedling development, xylem differentiation, or mitotic index in both lots. Cytogenetic analyses revealed that treatments with functionalized-HNT significantly increased the number of aberrations in low-vigor seeds. These results suggest that low-vigor seeds represent a model for a stress test that would be useful to monitor the effects of NPs. Moreover the present study offers scientific evidence for the use of halloysite for environmental purposes, supporting the biological safety of HNT-PdNPs. Environ Toxicol Chem 2016;35:2503-2510.

Ecotoxicity of halloysite nanotubes supported palladium nanoparticles on Raphanus sativus L.

Halloysite nanotubes (HNTs) are natural nanomaterials that are biocompatible and available in large amounts at low prices. They are emerging nanomaterials with appealing properties for applications like support for metal nanoparticles (NPs). The potential environmental impacts of NPs can be understood in terms of phytotoxicity. Current research has been focusing on HNT applications in cell or animal models, while their use in plants is limited so their ecotoxicological impact is poorly documented. To date there are no studies on the phytotoxic effects of functionalized halloysites (functionalized-HNTs). To develop a quantitative risk assessment model for predicting the potential impact of HNT-supported palladium nanoparticles (HNT-PdNPs) on plant life, an investigation was undertaken to explore their effects on seed germination, seedling development, and mitotic division in root tip cells of 2 lots of Raphanus sativus L. with different vigor. The results showed that exposure to 1500 mg/L of HNTs, functionalized-HNTs, and HNT-PdNPs had no significant influence on germination, seedling development, xylem differentiation, or mitotic index in both lots. Cytogenetic analyses revealed that treatments with functionalized-HNT significantly increased the number of aberrations in low-vigor seeds. These results suggest that low-vigor seeds represent a model for a stress test that would be useful to monitor the effects of NPs. Moreover the present study offers scientific evidence for the use of halloysite for environmental purposes, supporting the biological safety of HNT-PdNPs. Environ Toxicol Chem 2016;35:2503-2510.

Tocopherol, fatty acid and phytosterol content in seeds of nine wild taxa of Sicilian Brassica (Cruciferae)

Abstract The purpose of this study was to determine tocopherol (Toc), fatty acid and phytosterol content in dry seeds of nine wild taxa of Sicilian Brassica sect. Brassica. Analyses were performed by gas chromatography (GC)‐mass spectroscopy (MS). The biochemical analysis of dry seeds showed that in the nine taxa examined, α‐ and γ‐Toc were the major isomers, δ‐Toc was present in traces and β‐Toc was never found. All taxa had a similar fatty acid composition with C 16:0 (357.2 mg kg−1 DW), C 18:1 (393.94 mg kg−1 DW) and C 22:1 (618.29 mg kg−1 DW) acids present in higher amount. Unsaturated fatty acids (52.5–66.5%) were more abundant than saturated (33.5–47.5%) ones relative to the total fatty acids. Moreover, sitosterol was the most abundant (61.7–69.6%) of all phytosterols in all species and subspecies, followed by brassicasterol and campesterol at 16.0–19.3% and 11.9–20.7%, respectively. Total Toc content and the relative percentage of single isoforms were useful to discriminate seeds of B. incana, B. macrocarpa, B. rupestris group and B. villosa group. On the basis of these results, B. villosa subsp. villosa, B. villosa subsp. bivoniana, B. villosa subsp. drepanensis and B. villosa subsp. tinei can be ascribed to the B. villosa group. These taxa, having the highest T‐Toc, γ‐Toc and phytosterol contents, are potentially the most bioactive ones, and could, therefore, be exploited in breeding programmes to develop genotypes with putative positive effects on human health. Abbreviations: B, Brassica; DW, dry weight; FA, fatty acids; RH, relative humidity; T‐FA, total fatty acids; T‐Phy, total phytosterols; T‐Toc, total tocopherol; Toc, tocopherol

Assessment of genetic variation in Sicilian Helichrysum (Asteraceae) and implication to germplasm conservation

Abstract Some taxa of Sicilian Helichrysum related to sect. Stoeachadina have an uncertain systematic position; moreover, they are rare and endangered. In order to preserve these taxa in a genebank, a strategy to keep their genetic variation as high as possible is required. The high informative content of AFLP markers was used to assess genetic variations within and between populations and their relationships. The results show that all populations are genetically similar, even though there are two distinct groups: the first is the population referred to H. pendulum, and the second is represented by other populations belonging to several different specific and intraspecific taxa. The H. pendulum population is homogeneous, and characterised by the lowest genetic variation among all the populations considered. The second group shows a greater variation as evidenced by the presence of three subgroups: (i) populations referred to H. panormitanum var. panormitanum and H. panormitanum var. stramineum, (ii) populations referred to H. errerae var. errerae and H. errerae var. messerii, (iii) a population referred to H. hyblaeum, which, despite morphological and chorological differences, is similar to the other populations of the second group from a genetic stand point. Each population of the above-mentioned taxa has local fragment markers whose frequency is useful to determine how many plants should be sampled in order to collect the seeds necessary to preserve the populations genetic diversity in a genebank.

Chemical Composition of the Essential Oils of Three Endemic Species of Anthemis Sect. Hiorthia (DC.) R.Fern. Growing Wild in Sicily and Chemotaxonomic Volatile Markers of the Genus Anthemis L.: An Update

The chemical composition of the essential oils isolated from the aerial parts of Anthemis pignattiorum Guarino, Raimondo & Domina and A. ismelia Lojac. and the aerial parts and flowers of Anthemis cupaniana Tod. ex Nyman, three endemic Sicilian species belonging to the section Hiorthia, was determined by GC‐FID and GC/MS analyses. (Z)‐Muurola‐4(14),5‐diene (27.3%) was recognized as the main constituent of the A. pignattiorum essential oil, together with isospathulenol (10.6%), sabinene (7.7%), and artemisyl acetate (6.8%), while in the oil obtained from the aerial parts of A. ismelia, geranyl propionate (8.8%), bornyl acetate (7.9%), β‐thujone (7.8%), neryl propionate (6.5%), and τ‐muurolol (6.5%) prevailed. α‐Pinene was the main compound of both the aerial part and flower oils of A. cupaniana (18.4 and 13.2%, resp.). Also noteworthy are the considerable amounts of artemisyl acetate (12.7%) and β‐thujone (11.8%) found in the oil from the aerial parts and those of tricosane (9.8%) and sabinene (7.6%) evidenced in the flower oil. Furthermore, an update on the main compounds identified in the essential oils of all the Anthemis taxa studied so far was presented, and cluster analyses were carried out, to compare the essential oils of these taxa.

Stress integrated tests and cytological analyses reveal Brassica villosa subsp. drepanensis seed quality decrease upon long-term storage

Under stress integrated germination test (SIGT), seeds undergo osmo-saline stresses, which enable to detect differences in vigour of long-term stored seeds with high germination percentage (G%). The quality of Brassica villosa subsp. drepanensis seeds stored in a genebank (at − 20°C for 16 years) was compared with seeds at harvest by standard germination tests (GT), SIGT and cytogenetic analysis. No differences were detected in G% and mean germination time under GT. Conversely, SIGT performed with NaCl − 0.9 MPa osmotic potential did not influence G% at harvest but reduced that of stored seeds, SIGT at − 1.4 MPa reduced G% of both. Cytogenetic analysis showed reduction of mitotic index, appearance of chromosomal aberrations and smaller nucleoli in stored seeds compared with harvest seeds germinated in water. SIGT at − 0.9 MPa had no effect on mitotic index, but increased chromosome aberrations and nucleoli number. SIGT at − 1.4 MPa inhibited G% of harvest and stored seeds, reduced mitoses in harvest and completely prevented it in stored seeds. The results indicate that GT does not faithfully reflect the quality of stored seeds, with misinterpretation of their vigour, whereas SIGT and cytogenetical parameters are sensitive, reliable and inexpensive methods for early prediction of genetic erosion in germplasm banks.

Evaluation of the DNA barcoding approach to develop a reference data-set for the threatened flora of Sicily

The Mediterranean Basin is one of the most significantly altered World Biodiversity Hotspots with extensive habitat loss and fast genetic population erosion, for which urgent biodiversity reconnaissance and preservation actions are required. In particular, Sicily has about 600 taxa classified as threatened or near-threatened. The correct recognition and identification of such biodiversity is required for supporting further activities. The objective of this work is to assess the ability of the DNA barcoding approach to identify different taxonomic groups from a collection of the most threatened plant taxa, throughout natural Sicilian populations. The evaluation of the DNA barcoding core markers, rbcL and matK, was carried out on 30 taxa belonging to 13 families. DNA barcode fragments were recovered from all taxa (100%). The rbcL gene was recovered from 97% of the taxa and matK gene from 73%. In this test, 19 taxa overall (63%) were totally resolved at the specific or subspecific level, by at least one of the core markers. Fourteen of the 17 most threatened taxa (EN, CR) included in this work were totally discriminated. The matK and rbcL locus, respectively, resolved 64% and 48% of the taxa successfully sequenced. The matK gene expressed the highest genetic distance (K2P value), from 0.4% to 8.6%, against a range of 0.1–2% of rbcL gene. However, the rbcL gene appeared a good compromise between PCR, sequencing success and species-level resolution. Cryptic groups suggest the implementation of additional barcoding markers or different primer combinations, particularly for matK, in order to increase the performances. However, this preliminary result confirms the potential of the barcoding approach for quick identification of unknown and heterogeneous plant groups to generate a dedicated reference data-set of the threatened Sicilian flora for a wide range of applications.

Microwave effects on germination and growth of radish (Raphanus sativus L.) seedlings

Abstract Germination of radish seeds was delayed and reduced by low-power microwave exposure at 10.5 and 12.5 GHz. Irradiation decreased also hypocotyl growth rate. These effects were increased by rising of the microwave power and by the vertically polarised electromagnetic field. When the irradiation was suspended the seedling growth recovered.

Seed cryopreservation of Fraxinus angustifolia Vahl

Fraxinus angustifolia is used for afforestation and for production of both valuable timber and manna , a substance with pharmacological applications. The aim of this research was to establish the optimized condition for cryopreservation of F. angustifolia seeds, in order to protect the germplasm of this species. Germination percentage and mean time of germination of non-treated seeds (control) and liquid nitrogen-treated seeds were established in water or in 10 -6 M gibberellic acid (GA 3 ). The seeds could be cryopreserved with 3% of moisture content (MC) and germinated easily in water (70.0 ± 5.0%), while seeds cryopreserved with 6% MC showed a physiological dormancy. This state could be efficiently removed by addition of GA 3 , which increased germination to 76.0 ± 2.4%. Liquid nitrogen did not impair embryo viability, allowing a successful cryopreservation of seeds. The results are particularly useful for both germplasm conservation programs and in narrow leaved ash nurseries. Keywords : Fraxinus angustifolia , germination, germplasm conservation, seed cryopreservation African Journal of Biotechnology Vol. 12(16), pp. 1930-1936