Molecular genetic characterisation of the Asc locus of tomato conferring resistance to the fungal pathogen Alternaria alternata f. sp. lycopersici

The Alternaria stem canker disease of tomato is caused by the fungal pathogen Alternaria alternata f. sp. lycopersici and its host-selective AAL-toxins. Resistance to the pathogen and insensitivity to the toxins are conferred by the Asc locus on chromosome 3L. Sensitivity to AAL-toxins is a relative character; the toxins inhibit development of all tested tomato tissues but susceptible cultivars are much more sensitive than resistant cultivars. In addition to tomato, some other plant and animal species are sensitive to the toxins as well. The likely mode of action of AAL-toxins is interference with sphingolipid biosynthesis by specific inhibition of ceramide synthase activity. To molecularly isolate Asc, transposon tagging and positional cloning strategies are applied. As a first step, transposon insertions and restriction fragment length polymorphism (RFLP) markers are identified in proximity of the Asc locus. Subsequently, the transposons are used to inactivate Asc by insertion mutagenesis, and the RFLP markers are used to identify yeast artificial chromosomes (YACs) with tomato DNA inserts. Once an Asc-insertion mutant and/or a YAC encompassing Asc has been obtained, physical isolation and characterisation of Asc will be conceivable. Elucidation of the molecular role of Asc will illuminate the specificity of host recognition by Alternaria alternata f. sp. lycopersici.Abbreviations AAL-toxin Alternaria alternata lycopersici-toxin - A. a. lycopersici Alternaria alternata f. sp. lycopersici - Asc Alternaria stem canker - HST host-selective toxin     

Chromosome localisation of genes for resistance to Heterodera schachtii, Cercospora beticola and Polymyxa betae using sets of Beta procumbens and B. patellaris derived monosomic additions in B. vulgaris

Beet cyst nematodes (BCN, Heterodera schachtii), Cercospora beticola, and rhizomania, caused by the beet necrotic yellow vein virus (BNYVV) and vectored by the soil-borne fungus Polymyxa betae, are the most serious diseases of sugar beet ( Beta vulgaris subsp. vulgaris). The wild Beta species of section Procumbentes are known to be completely resistant to H. schachtii, C. beticola and P. betae. Alien monosomic additions (2n=19), plants of cultivated beet (2n=18) carrying different individual chromosomes of B. procumbens (2n=18) or B. patellaris (2n=36), were tested in greenhouse experiments for resistance to these pathogens. Gene(s) conferring full resistance to the beet cyst nematode in B. patellaris are located on chromosome 1.1, and the other tested chromosomes of B. patellaris are not involved in the expression of resistance. Artificial inoculation under greenhouse conditions, with in vitro produced inoculum of C. beticola and spot-percentage rating of the disease intensity, showed that the high level of resistance that was observed in the wild species B. procumbens and B. patellaris was not found in any of the monosomic additions tested. It was suggested that genes on various chromosomes of the wild species are needed to express full resistance, and that the chromosomes of group 7 of B. patellaris and chromosome 7 of B. procumbens have the largest effect. The greenhouse tests for resistance to P. betae in B. patellaris derived monosomic additions showed that the addition families of group 4.1 have a strong partial resistance, while the addition families of group 8.1 appeared to be completely resistant to the pathogen. Resistance to P. betae in the two wild species as well as in the two resistant addition types did not exclude infection with BNYVV, but resulted in a considerable reduction of the virus concentration. It was concluded that resistance to the vector would complement virus resistance, and may provide a more effective and durable control of rhizomania. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effects of Nitrate:Ammonium Ratios on Vegetative Growth and Mineral Element Composition in Leaves of Apple

Evaluations of vegetative growth and leaf concentrations of nitrogen (N), potassium (K), calcium (Ca), magnesium (Mg), and iron (Fe) were made of apple (Malus domestica Borkh. cvs. Granny Smith, Gala, and Golab) grown with five treatments of NO₃^?:NH₄⁺ ratios in pot culture. The concentrations of NO₃^?:NH₄⁺ ratios were 2.5:0.1, 6:0.3, 6:0.5, 6:0.7, and 6: 1 meq L^?1. Regression analysis showed that growth parameters of main stems and branches were not affected by increases of NH₄⁺ in the ratios. Granny Smith, Gala, and Golab differed in some of these parameters. Concentrations of N and Fe increased as NH₄⁺ increased, whereas K and Ca decreased and Mg was not affected significantly. Generally, the treatment of 2.5:0.1 produced leaves with lower N but higher K, Ca, and Mg concentrations than the other treatments. This research showed that vegetative growth was not affected by NH₄⁺ concentration whereas elemental composition was affected.     

Oocyte ultrastructural characteristics in camel (Camelus dromedarius) primordial to large antral follicles

This study was conducted to describe in detail the ultrastructural features and morphological characteristics of camel oocytes from preantral follicles in relation to the sequential stages of follicular development and also for oocytes from antral follicles in relation to their diameter. Camel oocytes from primordial, primary, secondary and also early to late antral follicles were processed and examined by light and transmission electron microscopy. Primordial follicular oocytes were characterized by a layer of flattened granulosa cells around and also eccentric nucleus and few cytoplasmic organelles in the peripheral region. Up to the secondary follicle stage, flat cells were replaced by cuboidal granulosa cells and their number increased and also an increase in the number of organelles such as vesicles, mitochondria and endoplasmic reticulum was observed. In the early antral stage, the formation of zona pellucida, appearance of microvilli and pleomorphic mitochondria was seen and the nucleus was dislocated to the peripheral region. During final growth phase, the extent of endoplasmic reticulum, vesicles and mitochondria increased, the number of lipid droplets decreased and cumulus cell process endings (CCPE) were observed. In conclusion, the growth of camel oocyte is associated with progressive increase in the number of mitochondria, endoplasmic reticulum, Golgi complexes and cytoplasmic vesicles as well as decrease in the number of lipid droplets and the nucleus migration from an eccentric in preantral to a peripheral location in antral follicles.     

Cumulus cell expansion and first polar body extrusion during in vitro oocyte maturation in relation to morphological and morphometric characteristics of the dromedary camel ovary

The morphological and morphometric characteristics of the ovary are fundamental properties for in vitro oocyte maturation. Nuclear maturation, including first polar body (1PB) extrusion, cytoplasmic maturation and cumulus cell (CC) expansion are the criteria for in vitro maturation (IVM) of oocyte. This study was designed to determine the effect of morphological and morphometric features of the ovary on CC expansion and 1PB extrusion during IVM of oocyte in the adult female dromedary camel. The weight, volume and three dimensions of ovaries from slaughtered dromedary camels and oocytes inside zona diameter and zona pellucida thickness were measured. The follicles were classified in regard to the size and oocytes according to their ooplasm appearance and CC compactness. Aspirated cumulus oocyte complexes (COCs) were incubated for 48 hr (with a 6‐hr interval) in Hams‐F10, and CC expansion and 1PB extrusion were assessed. Significant differences were seen in the shape, weight, volume and three dimensions of the ovaries between ≤4‐year‐old and >4‐year‐old dromedary camel (p < .5). Approximately, 95.82% of follicles were 2–4 mm in diameter. The mean (±SD) of inside zona diameter of the oocyte and zona pellucida thickness was 132.22 ± 13.8 and 14.64 ± 2.24 μm, respectively, in >4‐year‐old dromedary camel. The CC expansion and 1PB extrusion were seen in 86% and 21.88% of COCs, respectively. Age and sexual conditions of dromedary camel influence the morphological and morphometric characteristics of the ovary. Most COCs retrieved from 2–6 mm follicles are cultivable. The most slaughterhouse‐derived COCs retrieved from 2–6 mm follicles of non‐pregnant dromedary camels are excellent and good and yielding a most favourable diameter to achieve the developmental competence for IVM in an optimal time of 24–30 hr; the optimal time for CC expansion is 24–30 hr in this species. However, the CC expansion is a prerequisite process, but not sufficient for IVM.     

Integrated control of the solitary carpenter worm,Paropta paradoxa H.-Schaeff. (Lep., Cossidae) on fig trees in Egypt

According to the new trend of insect pests control in fruit orchards, biological in addition to mechanical control were evaluated for their efficiency on the reduction ofP. paradoxa population on fig trees during 1996, 1997 and 1998. Primary trials using the bacterial biopreparations of Delfin, Dipel and Bactospeine at the concentration of 200g/100 l. w. resulted in 81.8, 75.8 and 72.3% reduction of pest infestation, respectively. Pruning the infested branches, worming the larvae inside their tunnels and both mechanical treatments together along two successive years, reduced the infestation by 17.5, 47.4 and 57.9%, respectively. A control programme ofP. paradoxa was conducted using the techniques of pruning and worming besides spraying with bacterial insecticides at a rate of 200g/100 l.w. The performed mechanical and biological treatments together for one year gave 79.2–94.5% reduction of infestation, that reduction increased to 83.0–98.1% when these treatments were applied two years successively.     

Integrated control of the solitary carpenter worm,Paropta paradoxa H.-Schaeff. (Lep., Cossidae) on fig trees in Egypt

According to the new trend of insect pests control in fruit orchards, biological in addition to mechanical control were evaluated for their efficiency on the reduction ofP. paradoxa population on fig trees during 1996, 1997 and 1998. Primary trials using the bacterial biopreparations of Delfin, Dipel and Bactospeine at the concentration of 200g/100 l. w. resulted in 81.8, 75.8 and 72.3% reduction of pest infestation, respectively. Pruning the infested branches, worming the larvae inside their tunnels and both mechanical treatments together along two successive years, reduced the infestation by 17.5, 47.4 and 57.9%, respectively. A control programme ofP. paradoxa was conducted using the techniques of pruning and worming besides spraying with bacterial insecticides at a rate of 200g/100 l.w. The performed mechanical and biological treatments together for one year gave 79.2–94.5% reduction of infestation, that reduction increased to 83.0–98.1% when these treatments were applied two years successively.     

The effect of nitrate and plant size on nitrate uptake and in vitro nitrate reductase activity in strawberry (Fragaria × ananassa cv. Selva)

Nitrogen plays a major role in the growth and yield of strawberry. For optimizing nitrogen fertilizer application, it is necessary to understand the response of strawberry to nitrogen supply. The objective of this study was to determine the effect of nitrate supply and plant size on strawberry nitrate uptake and nitrate reductase activity (NRA). Strawberry plants cv. Selva were grown under growth chamber conditions in nutrient solutions containing 0, 0.1, 0.25, 0.7 or 4 mM nitrate. Cumulative nitrate uptake increased with the increase in nitrate supply. Increasing nitrate concentrations from 0 to 0.25 mM decreased leaf NRA; however, further increases to 4 mM nitrate restored NRA to activities observed at 0 mM nitrate. The activity of NADH- and NADPH-dependent NR was similar at every external nitrate concentration. Activity of NR was greater in the smallest plants and decreased as plant size increased. However, increasing external nitrate concentration increased nitrate uptake, but had not the same effect on nitrate reductase activity, showing that NRA and nitrate uptake are not necessarily correlated.     

Light and transmission electron microscopy of immature camelus dromedarius oocyte

In order to provide a consistent system for laboratory production of embryos, the characteristics of immature camel oocyte must first be described. The objective of this study was to define ultrastructural features of immature camel oocyte. Ovaries were obtained from camels at a local abattoir, and then transported to the laboratory within 2 h. Camelus cumulus oocyte complexes (COCs) were aspirated from 2-6 mm follicles using a 22-gauge needle. Excellent and good quality COCs were selected and prepared for transmission electron microscopy study using a cavity slide. The fine structure of camel oocyte is morphologically similar to that of other mammalian oocytes. However, some minor differences exist between COC of camel and other mammalian species. Different size and shape of membrane-bound vesicles, lipid droplet, mitochondria and cortical granules were distributed throughout the ooplasm. Discrete or in association with endoplasmic reticulum, Golgi complexes were observed in the periphery of the oocytes. The majority of the oocytes were in the germinal vesicle stage.