Quantifying the relationship between disease severity and the amount of Aphanomyces euteiches detected in roots of alfalfa and pea with a real-time PCR assay

A real-time PCR assay was used to quantify the relationship in alfalfa and pea between disease severity and the amount of Aphanomyces euteiches detected in roots. The study included isolates of race 1 and race 2 of the alfalfa pathovar of A. euteiches and an isolate obtained from diseased pea. Spearman rank correlations between pathogen DNA content and disease severity index (DSI) ratings were positive ( ? 0.57) and significant (P  0.0007) for individual alfalfa plants, bulked alfalfa plant samples, and individual pea plants. In all experiments, significantly more pathogen was detected in susceptible populations than in resistant populations. The results clearly demonstrate that resistance to A. euteiches in both alfalfa and pea is characterized by a reduction in pathogen colonization relative to levels observed for susceptible reactions. The assay was very specific for A. euteiches, producing very linear assays with DNA extracted from pathogen isolates obtained from alfalfa, pea, and bean. Possible applications of the assay in conjunction with other real-time PCR assays specific to other legume pathogens are discussed in relation to simultaneous disease screening for multiple plant pathogens and the study of microbial population dynamics in mixed plant infections.     

Eleven ancestral gene families lost in mammals and vertebrates while otherwise universally conserved in animals

Background  

Gene losses played a role which may have been as important as gene and genome duplications and rearrangements, in modelling today species' genomes from a common ancestral set of genes. The set and diversity of protein-coding genes in a species has direct output at the functional level. While gene losses have been reported in all the major lineages of the metazoan tree of life, none have proposed a focus on specific losses in the vertebrates and mammals lineages. In contrast, genes lost in protostomes (i.e. arthropods and nematodes) but still present in vertebrates have been reported and extensively detailed. This probable over-anthropocentric way of comparing genomes does not consider as an important phenomena, gene losses in species that are usually described as "higher". However reporting universally conserved genes throughout evolution that have recently been lost in vertebrates and mammals could reveal interesting features about the evolution of our genome, particularly if these losses can be related to losses of capability.     

Single-molecule anisotropy imaging

A novel method, single-molecule anisotropy imaging, has been employed to simultaneously study lateral and rotational diffusion of fluorescence-labeled lipids on supported phospholipid membranes. In a fluid membrane composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine, in which the rotational diffusion time is on the order of the excited-state lifetime of the fluorophore rhodamine, a rotational diffusion constant, D(rot) = 7 x 10(7) rad(2)/s, was determined. The lateral diffusion constant, measured by direct analysis of single-molecule trajectories, was D(lat) = 3.5 x 10(-8) cm(2)/s. As predicted from the free-volume model for diffusion, the results exhibit a significantly enhanced mobility on the nanosecond time scale. For membranes of DPPC lipids in the L(beta) gel phase, the slow rotational mobility permitted the direct observation of the rotation of individual molecules characterized by D(rot) = 1.2 rad(2)/s. The latter data were evaluated by a mean square angular displacement analysis. The technique developed here should prove itself profitable for imaging of conformational motions of individual proteins on the time scale of milliseconds to seconds.     

Summer diapause and winter quiescence ofCoccinella septempunctata (Col. Coccinellidae) in central Greece

Coccinella septempunctata (Coleoptera: Coccinellidae) in Greece is a multivoltine species. In the lowlands, all instars are abundant in spring becoming scarce from July until the end of the warm period of the year; they are absent in winter. In June, mostC. septempunctata adults migrate to mountainous aestivo-hibernation sites. Measurements of the duration of pre-oviposition period in females taken monthly from the summit of Mount Kitheron in central Greece (1993–1994) and transferred to laboratory conditions of high temperature (25°C), long day (16 hrs light/24 hrs), and presence of aphids, indicated that theC. septempunctata females were in diapause during July and August. The diapause gradually terminated from late August to late October and was followed by a period of quiescence extending from November to March of the following year. During the summer diapause, arrest of ovarian development was indicated by immaturity of the ovaries in all sampled females and the complete absence of vitellogenic resorption signs. Also, adults were found with enlarged fat bodies, and the median duration of preoviposition period in females transferred to the above laboratory conditions was 90 days in early July and 82 days in early August. During the period of winter quiescence, arrest of ovarian development was characterized in most samples by immaturity of the ovaries in all females and, in some samples, by the presence of a few females with signs of vitellogenic resorption. In winter, adults were found containing fat body reserves of different levels, and the median preoviposition period of females transferred to optimal breeding conditions was 29 days in early November and 16 days in mid January.     

Phenology ofCoccinella septempunctata (Col.: Coccinellidae) in central Greece

Coccinella septempunctata (Coleoptera: Coccinellidae), field collected from central Greece and reared in cages during 1990–1991 and in vials during 1994–1995 outdoors at Kifissia, Athens, developed 4 complete and a 5^th partial generations per year. In both cases, only adults of the 1st generation reproduced within the year they emerged; 2^nd, 3^rd and 4^th generation adults reproduced partly in the year they emerged and partly in the following year; 5^th generation adults did not reproduce at all in 1990, and eggs/larvae produced by 4^th generation females in 1994 died before developing into adults. Adults of the 1 st and 5th generations died before winter; those of the 2^nd, 3^rd and 4^th generations overwintered successfully. The greatest numbers of eggs were laid by females of the 1^st and 2^nd generations. Visual counts made in the open field at Kopais plain in central Greece (1990–1992) and on the summit of the adjacent Mount Kitheron (1990–1994) indicate that allC. septempunctata instars were abundant in the plain between April and June, becoming scarce from July until the end of the warm period of the year following spring. NoC. septempunctata individuals were found in the plain in winter. On the summit of the mountain,C. septempunctata adults were present all year round, singly and in aggregations, except for the month of May. The most numerous arrivals of adults were noticed on the mountain in June. Emigrations of adults from there were noticed from March until the end of April.     

Phenology ofHippodamia undecimnotata (Col.: Coccinellidae) in Greece

Hippodamia (Semiadalia) undecimnotata (Coleoptera: Coccinellidae), collected from central Greece and reared in cages during 1993–1994 and in vials during 1994–1995 outdoors at Kifissia, Athens, completed 5 generations per year. In both cases, adults of the 1^st, 2^nd and 3^rd generations reproduced, completing their egg laying in the same year; 4^th and 5^th generation adults reproduced both in the year they emerged and the following year. In both cases, adults of the 1^st and 2^nd generations died before winter; in 1993, adults of the 3^rd generation died before winter, in 1994, they survived until April 1995. Adults of the 4^th and 5^th generations overwintered successfully. The greatest numbers of eggs were laid by females of the 1^st and 2^nd generations. Field observations and timed counts of specimens (30 minutes per location) made on the tops of Mounts Chlomo and Kitheron and on the neighboring plain of Kopais between 1991 and 1994 revealed that most adults arrived on the mountains between late June and early September and left between the end of March and beginning of May.H. undecimnotata were not found on the plain of Kopais during the winter. Overwintered adults arrived between the end of March and beginning of May; 1^st generation adults emerged in June. The presence of a few eggs, larvae, pupae and newly-emerged adults observed on the plain during August-September 1993 suggests that at least a portion of theH. undecimnotata population in central Greece completes 2 or more overlapping generations per year.