In vitro development and germination of immature olive embryos

Summary

In vitro culture methods were used to germinate olive embryos prior to maturation. Fruit, seed and embryo development were established with consecutive sampling from 20 to 100 days after bloom. For that same period, embryo development and germination success were determined by in vitro culture trials using one-third strength MS medium with or without the addition of zeatin. For early developmental stages, when isolation of the embryo was difficult, a cut portion of the seed containing the embryo was used for culture. The embryos cultured within the cut seed portions germinated and formed normal plantlets. Histological observations indicated a close similarity between the natural and in vitro immature embryo differentiation pattern, progressing through preglobular, globular, heart-shaped and torpedo-shaped stages. In some cases, however, the in vitro immature embryos developed or germinated abnormally. The presence of zeatin (0.25 mg l^–1) in the culture medium and the use of a cut seed-portion containing the immature embryo allowed in vitro germination sooner after bloom than previously obtained. On the contrary, zeatin was a handicap for mature olive embryo in vitro germination, which reached 100% seedling formation when no plant growth regulators were used.     

Productivity of olive trees with different water status and crop load

SUMMARY

A field experiment was conducted over two growing seasons to determine the combined effect of crop load and irrigation on yield components of olive trees (Olea europaea L. ‘Leccino’) planted at 6 m 3.8 m in a sandy-clay soil. Different crop loads were established by manual thinning of fruits. Drip irrigation was managed to maintain pre-dawn leaf water-potentials (PLWP) within the following ranges: (i) higher than –1.1 MPa (FI; fully irrigated); (ii) between –1.0 and –3.3 MPa (DI; deficit irrigated); or (iii) below –1.2 MPa, but not lower than –4.2 MPa (SI; severe deficit irrigated). The irrigation period lasted from 6 – 16 weeks after full bloom (AFB) in 2003, and from 5 – 19 weeks AFB in 2004. In 2003, full bloom was on 26 May; in 2004, it was on 3 June. Neither irrigation regime nor crop load affected flowering or flower quality the following Spring. The combined fruit yields [on a fresh weight (FW) basis] over both years in SI and DI trees were 49.0% and 81.6% of FI trees, respectively. The oil yields of SI and DI trees were 52.5% and 81.2% of FI trees, respectively. Fruit FWs in FI trees were greater than those of DI or SI trees at 8 weeks AFB. At harvest, FI trees bore the largest fruits, and SI trees the smallest fruits. The FWs of individual fruits at harvest in the FI and DI treatments decreased as crop load increased, but no such relationship was apparent for SI trees. The oil content of the mesocarp increased as PLWP increased from approx. –3.5 MPa to –1.5 MPa. The oil content of FI trees at harvest decreased from 53.1% to 45.7% dry weight as fresh fruit yield increased from 5 – 25 kg dm^–2 trunk cross-sectional area. However, crop load did not have any effect on the oil content of the mesocarp in DI trees. Fruit maturation was delayed by irrigation. Maturation index also decreased (indicating delayed maturation) as the crop load on FI or DI trees increased, but did not vary with crop level in SI trees.     

Olive seedling first-flowering position and management

The prolonged length of the juvenile period represents a substantial obstacle in olive (Olea europaea) breeding programs, delaying both the possibility for analyzing the fruit, the harvest unit, and the capacity for sexual recombination. In both olive and other fruit tree species, the juvenile–adult transition has been successfully hastened by forcing and formation procedures designed to rapidly achieve a minimum height. Precise knowledge of the position within the tree canopy where the juvenile–adult transition occurs, identified by the location of the first flowers, offers further potential for manipulating tree structure in order to hasten that transition. The occurrence of the juvenile–adult transition has been described spatially within the canopy as the formation of a juvenility cone, which we report here for olive plants from seed, and for the first time quantitatively describe its position based on the added trunk and branch distances from the soil. In canopies of seedlings from open pollination of cv. Arbequina olive the first-flowering position was consistently located, in our conditions, at an average distance of 200 cm from the trunk base. Based on that evidence, that first flowering can occur at lower positions, and on the values we obtained, three canopy-formation heights (100–130 cm, 130–160 cm and >160 cm) were compared for their effect on the length of the juvenile period and the vigour of olive seedlings. Canopy heights of 100–130 cm, much lower than the 160 cm previously reported by the olive tree breeding program of Córdoba, produced the highest number of flowering plants in the first two flowering years and also provided easier management.     

Immunohistochemical detection of virus antigen in the nasal planum of rabid dogs

The rabies virus is one of the most neurotropic of all viruses infecting mammals. During the terminal phases of infection, the virus spreads to peripheral tissues, including the skin. The external skin of the nose, called the nasal planum, is a sensory organ where numerous nerve bundles and terminal nerves are distributed. Therefore, the nasal planum is expected to serve as a postmortem diagnostic material. However, the distribution of rabies virus antigens in the nasal planum in rabid animals has not yet been studied. In this study, the nasal planum was obtained from 45 rabid dogs. In all rabid dogs, the viral antigen was detected in the peripheral nerve tissues, Merkel cells, and squamous cells. The viral antigen in the epidermis exhibited three patterns: first, a diffuse positive pattern from the basal layer to the squamous layer; second, a reticular positive pattern along the cell membrane in the squamous layer; and third, a basal layer pattern of the epidermis. In the dermis, viral antigens were detected more often in lamellated corpuscles just beneath the rete pegs. These results suggest that the nasal planum could serve as a useful alternative source for postmortem diagnosis in rabies endemic countries.     

Oxygenated Acyclic Diterpenes with Anticancer Activity from the Irish Brown Seaweed Bifurcaria bifurcata

Brown alga Bifurcaria bifurcata is a prolific source of bioactive acyclic (linear) diterpenes with high structural diversity. In the continuation of our investigations on Irish brown algae, we undertook an in-depth chemical study on the n-hexanes and chloroform subextracts of B. bifurcata that led to isolation of six new (1–6) and two known (7–8) acyclic diterpenes. Chemical structures of the compounds were elucidated by a combination of 1D and 2D NMR, HRMS, FT-IR, [α]_D and vibrational circular dichroism (VCD) spectroscopy. Compounds 1–8, as well as three additional linear diterpenes (9–11), which we isolated from the same seaweed before, were tested against the human breast cancer cell line (MDA-MB-231). Several compounds moderately inhibited the growth of the MDA-MB-231 cell line with IC₅₀ values ranging from 11.6 to 32.0 μg/mL. The present study carried out on the lipophilic extracts of the Irish B. bifurcata shows the enormous capacity of this seaweed to produce a large palette of acyclic diterpenes with diverse oxygenation and substitution patterns and promising bioactivities.     

Genetic variability of Polish tomato black ring virus isolates and their satellite RNAs

Tomato black ring virus (TBRV), a member of the Nepovirus genus, is a serious plant pathogen distributed worldwide. It causes significant damage to several economically important crops, such as artichoke or strawberry. The TBRV bipartite genome consists of two polyadenylated single-stranded positive-sense RNA molecules, which may be accompanied by subviral particles such as defective interfering RNAs (DI RNAs) and satellite RNAs (satRNAs). In this study, we obtained the complete genome sequence of six TBRV isolates originating from different hosts and determined the presence of eight TBRV satRNAs. Subsequently, genetic variability, recombination, phylogenetic and selection pressure analyses were performed. The results revealed that the TBRV population is genetically diverse. The occurrence of potential recombination events, evidence of positive selection pressure acting on particular codons and the diversification of satRNAs within the TBRV population indicated that the virus mutates and can rapidly adapt to new environmental conditions or hosts. The presented data shed some light on TBRV evolutionary dynamics and epidemiology.     

Investigation of genome polymorphism and seed coat anatomy of species of section Adenolinum from the genus Linum

Twelve species of section Adenolinum from genus Linum have been studied using cytogenetic methods (DAPI/C-banding, Ag-NOR staining, FISH with 5S and 26S rDNA probes), RAPD analysis, and seed surface ultrastructure and seed coat anatomy investigation. Karyotype analysis revealed that these species have a chromosome number of 2n = 18 or 2n = 36. The DAPI/C-banding patterns enabled the identification of all chromosomes and revealed the similarities in chromosome patterns in all of the studied species. It was found that 18-chromosome plants are diploids, whereas 36-chromosome forms are tetraploids. In diploid species, 5S and 26S rRNA genes are located in similar positions on a single pair of chromosomes or on two chromosome pairs in tetraploids. Ag-NOR staining showed NOR activity in both genomes of tetraploids. RAPD analysis and seed coat anatomy investigation revealed some interspecies differences. Species from nearby areas have more similar RAPD characteristics compared with more geographically distant species. The results of this study may serve as an additional argument in favour of a monophyletic origination of the Adenolinum species and its strong isolation from other Linum species.