An In Vitro Test for Temperature Sensitivity and Resistance to Meloidogyne incognita in Tomato

An in vitro root explant tissue culture technique is described for determining susceptibility of tomato (Lycopersicon esculentum Mill.) breeding lines and cultivars to the root-knot nematode Meloidogyne incognita. Root explants were taken from 2-day-old seedlings cultured for 30 days at 28 C on Gamborg''s B-5 medium with or without nematode inoculum. The remaining portion of the root and stem from the excised root explants was transferred to soil in pots and grown to maturity in the greenhouse. In vitro root explants were evaluated for growth and occurrence of juveniles, adults, and egg masses. The regenerated plants were used to produce more seed, The proposed technique is simple, reliable, and adapted to routine screening of large numbers of F₁ and F₂ samples, and it utilizes less space than tests performed on intact plants in the greenhouse or growth chamber. Evidence is presented also on the breakdown of resistance to M. incognita under high temperature stress using this in vitro root explant technique.     

Survey of Heterorhabditidae and Steinernematidae (Rhabditida,Nematoda) in Western Canada

A survey was done in the summer months along the Alaska Highway, in other parts of British Columbia, in northern Alberta, and in the Yukon Territory for steinernematid and heterorhabditid nematodes occurring in the top 10 cm of soil. Steinernema feltiae and Steinernema spp. were found at 18 and Heterorhabditis megidis at 7 sites of 125 sampled. Most nematodes were found where visible insect infestation occurred and where human influence on the habitat was substantial (e.g., agricultural, forested and bush-hedgerow habitats); none was found in grassland or virgin forests. Heterorhabditis megidis occurred in only the southern, warmer, drier region of British Columbia. In the laboratory some steinernematid isolates and H. megidis killed Galleria mellonella larvae at 13 and 22 C, whereas some isolates of Steinernema killed the larvae at only 13 C. Steinernema spp. from three high altitude sites with low, average July temperatures (13-14 C) are cold-active in that they produced infective juveniles at 13 C and killed G. mellonella at 6 C.     

Effect of meteorological variates on persistence of primary conidia and capilliconidia of Erynia radicans (Zygomycotin: ntomophthorales) under natural conditions

Persistence of conidia of an isolate of Erynia radicans (Syn. Zoophthora radicans) was investigated in relation to the meteorological conditions which occurred during autumn-winter of 1990–91 in the coastal plain in Israel. Capilljconidia shielded from the sun, placed on the abaxial surface of leaves of Plumeria acuminata, persisted for 24 h to at least 120 h. Exposed capilliconidia, placed on the adaxial surface of the same leaves, died within 24 h. Almost all the primary conidia shielded from the sun (placed on the abaxial surface of the same leaves) died within a single day. Conidial viability was expressed in subsequent germination on an agar medium. Capilliconidial persistence was closely related to the daily air temperatures, expressed as cumulative day-degrees. Differences in relative humidity had no substantial effect on capilliconidial mortality. At daytime temperatures of ≤ 20°C, mortality after 24 h incubation was lowest (≤ 34%) and the persistence duration, longest (at least 120 h). Increases in daytime temperature up to 24°C for a few hours increased mortality (37–57% after 24 h incubation) and shortened the persistence duration (72–120 h). Exposure to 24–29°C during daytime greatly increased mortality (65–58% after 24 h) and further shortened the persistence duration (24–48 h). Daytime temperatures of > 29°C were lethal to all capilliconidia within 24 h. Temperature had a profound effect on capilliconidial persistence also under controlled environmental conditions. The significance of capiliiconidial persistence is discussed in relation to activity of the fungus in its natural environment.     

The influence of temperature on light enhanced glycoalkaloid synthesis in potato

The influence of temperature on total glycoalkaloid (TGA) synthesis in tubers exposed to light (250 jumol m“² s”² PAR, Photosynthetically Active Radiation) or dark environments for 96 h was examined in three potato cultivars. Following 96 h light or dark the tubers were stored without light at 5°C or 24°C and TGA concentrations monitored over the subsequent 30 and 90 days. Exposure to light and cultivar were found to be major factors influencing TGA concentrations; temperature had no significant effect. TGA content in illuminated tubers of cvs ‘Pentland Hawk’ and ‘Kerrs Pink’ were significantly higher (P < 0.01) compared with tubers placed in the dark. TGA concentrations in cv. ‘Desiree’ increased significantly only following exposure to light at low temperatures (P < 0.05). Removal of tubers from storage at 5°C and immediate illumination at 24°C altered the ratio of glycoalkaloids in cvs ‘Pentland Hawk’ and ‘Kerrs Pink’. Regardless of cultivar and storage temperature TGA concentrations were higher at the end of the storage period compared with initial TGA concentrations. During storage TGA concentrations fluctuated widely and gradual accumulation of glycoalkaloids with time was rarely demonstrated except in cv. ‘Desiree’. Tubers stored at 24°C accumulated higher TGA concentrations than those stored at 5°C in cv. ‘Kerrs Pink’ but not in cvs ‘Pentland Hawk’ and ‘Desiree’. Tubers of cv. ‘Kerrs Pink’ exposed to light prior to storage accumulated glycoalkaloids more rapidly than unexposed tubers during storage at 24°C and occasionally at 5°C. Light enhanced glycoalkaloids are not degraded over time.     

Issues in the culture of the extremely thermophilic methanogen, Methanothermus fervidus

Basic issues in the culture of the extremely thermophilic archaeon, Methanothermus fervidus, have been investigated, including culture medium formulation, substrate yield and product yield coefficient, growth rate and stoichiometry, and H(2) uptake kinetics. The pH optimum for growth of this organism was estimated at 6.9. Growth medium buffered with PIPES instead of bicarbonate supported both increased growth rate and maximum biomass concentration. Substitution of titanium(III) citrate for the reducing agent sodium sulfide improved culture performance as well. However, independent adjustment of iron and nickel concentrations from 11 to 111 muM, respectively, and carbon dioxide partial pressure from 5 to 20 psia did not impact the culture of M. fervidus significantly. An elemental balance approach was utilized to aid in design of a defined medium to support growth to a target maximum biomass concentration of at least 1.0 g dry wt/L. The growth of this organism was limited by H(2) availability in this reformulated culture medium. The maximum growth rate and biomass concentration achieved in anaerobic vials with the defined medium was 0.16 h(-1) and 0.74 g dry wt/L, respectively. This maximum biomass concentration was a 72% improvement over that obtained with a literature-based defined medium. The Monod parameter, K(s), with H(2) as limiting substrate, was estimated at 1.1 +/- 0.4 psia (55 +/- 20 muM in the broth), based on a H(2) consumption study. Representative values for the substrate yield, Y(X/CO(2) ), and product yield coefficient, Y(CH(4)/) (X), were determined experimentally to be 1.78 +/- 0.04 g dry wt/mol CO(2), and 0.52 +/- 0.01 mol CH(4)/g dry wt, respectively. A bench-scale fermentation system suitable for the culture of extremely thermophilic anaerobes was designed and constructed and proved effective for the culture of M. fervidus. (c) 1993 Wiley & Sons, Inc.     

Fed-batch cultivation of recombinant escherichia coli JM103 and production of the fusion protein SPA::EcoRI in a 60-L working volume airlift tower loop reactor

SPA::EcoRI fusion protein was produced by Escherichia coli JM103 carrying the multicopy expression plasmid pMTC48, the multicopy repressor plasmid pRK248, and the multicopy protection plasmid pEcoR4 in a 60-L working volume airlift tower loop reactor on M9 minimal medium with glucose. Cell mass concentration, total cell count, number of colony-forming units, specific growth rate, yield coefficient, and metabolite (acetate, pyruvate, succinate, lactate, ethanol) concentrations were monitored during the growth phase and gene expression. Gene expression was induced by temperature shift or chemically by isopropyl-thiogalactosidase in the airlift tower loop reactor (ALTR) at constant cultivation time and in a small stirred tank reactor at different cultivation times. During induction, the cultivation medium was supplemented with concentrated Luria-Bertani (LB) medium. The intracellular enzyme activity was evaluated as a function of the time after the start of the induction. It was found that the reduction of the glucose concentration and increase of the dissolved oxygen concentration reduced the acetate produced and increased the intracellular enzyme activity. (c) 1993 John Wiley & Sons, Inc.     

RESPONSES AND CORRELATED RESPONSES TO ARTIFICIAL SELECTION ON THORAX LENGTH IN DROSOPHILA MELANOGASTER

Two sets of four replicate lines of Drosophila melanogaster were selected for large and small thorax with controls. F, progeny of crosses between the selected lines within each size category showed (a) a reduction in preadult viability in large lines relative to control and small lines when they were cultured at medium or high density in competition with a standard mutant marked competitor stock, and (b) an increase in larval development time in large lines relative to control and small lines. Natural selection for increased body size in adults may therefore be opposed by adverse effects on larval viability. The results are discussed in terms of the developmental mechanisms probably responsible for the change in body size. The preadult survival of the large and control lines was measured at three different temperatures, and there was no evidence for a significant interaction between size and temperature. The observed evolutionary increase in body size in response to reduced temperature in Drosophila must therefore involve either different genes from those subject to selection for size at a single temperature, or a fitness component other than preadult survival. There was no significant asymmetry in response to selection, and thorax length showed heterosis in crosses between the selected lines.     

Temperature preferences and tolerances of three fish species inhabiting hyperthermal ponds on mangrove islands

The fish species Cyprinidon artifrons, Floridichthys carpio, and Gambusia yucatana inhabit shallow mangrove ponds off the coast of Belize. Portions of these ponds experience a diurnal temperature change from 26 °C at night to 40 °C and above during midday. Repeated field observations indicate Cyprinidon prefer the warmer (and much larger) portions of the ponds whereas the other two species stay in the cooler areas.The hypothesis that temperature is serving as a cue for partitioning within the ponds was supported by laboratory thermal gradient tests in which Cyprinidon preferred temperatures clearly higher than the other two species.The critical thermal maximum (CTM) was determined for the three species using members that had been acclimated to either a daily cycling temperature similar to that for the ponds, or to the mean of the 24-hour cycle (30 °C). Cyprinidon acclimated to the cycling temperature had a CTM of 45.5 °C, which apparently sets a new record for fish CTM. Acclimation to a constant 30 °C lowered the CTM to 43.7 °C. Floridichthys and Gambusia acclimated to the cycled temperature had CTMs of 43.9 and 43.3 °C respectively, and 42.5 and 42.6 °C for those acclimated to 30 °C.All three species appear to have the ability to tolerate the high temperatures throughout the ponds but only Cyprinidon utilize the whole pond during the day. This may help to explain the large populations of Cyprinodon found in these mangrove ponds compared to the other species.     

Low temperature compartment formation in feline immunodeficiency virus-infected and uninfected feline kidney cells

Summary This study was to determine if feline immunodeficiency virus (FIV)-infected and uninfected Crandall feline kidney (CRFK) cells exhibited a low temperature (16°C) block in membrane trafficking between transitional endoplasmic reticulum and Golgi apparatus represented by intermediate compartment formation. Cells were cultured at different temperatures and membrane changes involving the Golgi apparatus and Golgi apparatus-associated membrane structures were monitored by electron microscopy and quantitated. With 30 min of incubation, membranes of the Golgi apparatus stack increased in amount at temperatures of 16°C and below compared to temperatures above 18°C. The increase was greatest along the major polarity axis as evidenced by an increased stack height. Neither the number of cisternae per stack nor the average stack diameter (width) was affected by temperature. The response was maximal between 15 and 30 min of low temperature treatment of the cells. Results with cells infected and uninfected with feline immunodeficiency virus were similar. The increase in stack height was due primarily to an increase of membranes at the cis face (cis Golgi apparatus network). At 18°C, membranes of the trans Golgi apparatus network accumulated suggesting that import from the cis Golgi network could proceed at this temperature, whereas exit from the trans Golgi network was still at least partially blocked. Also increased at 16°C and below were numbers of transition vesicles in the space between the Golgi apparatus and the transitional endoplasmic reticulum associated with the cis Golgi apparatus face. The results suggested interruption of the orderly flux of membranes into the Golgi apparatus at 16°C and below. Moreover, the block appeared to be reversible. Upon transfer from 16°C to 37°C, there was a time-dependent decrease in the accumulations of cis compartment membrane accompanied by a corresponding equivalent increase in the membranes of the trans Golgi apparatus compartment.     

Effects of body and head positions on bilateral difference in tympanic temperatures

We have examined the nonparallel changes in tampanic membrane temperatures (T _ty) from the two ears in response to various changes in body and head positions. Upon assuming a lateral recumbent position, the T _ty on the lower side increased while that on the upper side decreased. Pressure application over a wide area of the lateral chest only caused inconsistent and obscure asymmetric changes in T _ty. A lateral flexion of the head with the subject sitting upright and a rotation of the head to the side in a supine position induced an increase in the T _ty on the lower side compared to that on the upper side. The temperature and blood flow of the forehead often decreased on the lower side and increased on the upper side, although such responses were not always concomitant with the asymmetric changes in T _ty. A dorsal flexion of the head with the subject in a reclining position caused a slight increase in the T _ty, whereas raising the head upright induced a slight decrease in them. Two additional experiments were carried out with single photon emission computed tomography using ^99mTc-hexamethylpropyleneamine oxime as tracer, and a slight, relative decrease in counts was noted in the right hemisphere during rotation of the head to the right. These results would strongly suggest that unilateral increases and decreases in T _ty could have been caused by one-sided decreases and increases, respectively, in blood flow to the brain and/or the tympanic membrane, induced by a vasomotor reflex involving vestibular stimulation.