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PubMed1993

Alpha-motoneuron excitability at high altitude.

It has been hypothesized that chronic hypobaric hypoxia could lead to inhibition of the alpha-motoneuron pool, thus limiting the maximal activation of working skeletal muscles. To test this hypothesis six subjects [32 (SEM 2) years] were evaluated in resting conditions, at sea level and after acclimatization at 5,050 m. The recruitment curves of the Hofmann-reflex (H-) and the direct muscle-response. (M-) of the right soleus muscle were obtained by stimulating the posterior tibial nerve with different intensities while recording the electromyogram of the soleus muscle. From the recorded data the net alpha-motoneuron excitability (ratio of maximal H-reflex to M-response Hmax:Mmax ratio), the threshold and gain for both responses, obtained from linear regressions through the rising phase of the recruitment curves of both responses, as well as the latency times of both responses were determined. The latency times and the Hmax:Mmax ratio were unchanged at altitude. The thresholds of both responses and the gain of the M-response were unaltered. The gain of the H-response was significantly higher at altitude when compared to sea level.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1991

Morphological changes in the pituitary-adrenocortical axis in natives of La Paz.

Increased activity of the hypothalamic-pituitary-adrenocortical axis is part of the response to the stress of initial exposure to hypoxia, but there is evidence to suggest that it persists after homeostatic stability has been regained and acclimatization achieved. The adrenal glands of five lifelong residents of La Paz, Bolivia, who had lived at altitudes in the range 3600-3800 m, were significantly larger than those in age-matched controls from sea level (15.3 g vs 10.4 g; P less than 0.001) and appeared hyperplastic. The pituitary glands of the highlanders were not significantly different in size from those of the controls (0.67 g vs 0.51 g), but contained larger populations of corticotrophs expressed in terms of the total cell population of their anterior lobes (25.6% vs 19.4%; P less than 0.001). In conjunction with other studies of this endocrine axis in man and animals exposed to a hypoxic environment, these data suggest that greater amounts of adrenocorticotrophic hormone (ACTH) are required to maintain normal adrenocortical function under such circumstances, probably as a result of hypoxic inhibition of adrenocortical sensitivity to stimulation. Physiological hyperplasia of the adrenal cortex may be common in people living at high altitude.

Adrenocorticotropic Hormone↗

PubMed1986

Temperature dependence of neurotransmitter release in the antarctic fish Pagothenia borchgrevinki.

The quantal contents of endplate potentials from extraocular muscles of an antarctic fish Pagothenia borchgrevinki were measured over a range of temperatures. Quantal release was maximal at about 5 degrees C but showed little dependence on temperature between -2 degrees C and 10 degrees C. Above 10 degrees C quantal content declined until release ceased about 18 degrees C. In view of the fact that the ambient temperature at which these fish live is constant at -1.9 degrees C, the results suggest that Pagothenia borchgrevinki is only partially adapted to its environment despite 25 million years acclimatization.

Animals↗

PubMed1996

Aerobic capacity and skeletal muscle properties of normoxic and hypoxic rats in response to training.

The aim of this study was to determine, in the rat, the effects of chronic exposure (7-9 weeks) to normobaric hypoxia (FIO2=0.13, equivalent to 3700 m altitude) on cardiac and skeletal muscle properties, on maximal oxygen uptake (VO2max), and endurance time to exhaustion (ETE). In addition, we evaluated the impact of endurance training (90 min of treadmill running per day, 5 days per week, for 9 weeks) on these parameters. The results were compared to normoxic rats fed ad libitum (NAL) and to normoxic pair-weight (NPW) animals in order to take into account the influence of hypoxia on growth rate. It was found that, in sedentary rats, hypoxia results in stunted growth, adrenal atrophy, a significant reduction of cross-sectional area of fast-twitch (type II) fibres, a reduced capillary-to-fibre ratio (C/F), and a reduced oxidative capacity (decreases in citrate synthase and 3-hydroxy-Acyl CoA dehydrogenase activities) of the plantaris muscle. These effects are mainly related to the anorexic effects of prolonged exposure to hypoxia. Nevertheless, hypoxic (H) rats displayed higher VO2max and ETE values when compared either to NAL or to NPW animals. Endurance training resulted, in all groups (H, NAL, NPW), in a significant change of the fibre type distribution of the plantaris which displayed an increased number of type IIA fibres and a decreased proportion of type IIB fibres. In addition, the C/F ratio and cross-sectional area of fast-twitch fibres were normalized by superimposition of training on hypoxia. Both VO2max and ETE were significantly higher in trained H rats than in NAL, but these improvements were mainly related to the reduced body weight induced by hypoxia. These data suggest that the greater aerobic capacity and tolerance for prolonged exercise induced by chronic exposure to hypoxia can be mainly accounted for by the anorexic effects of hypoxia, although other factors (e.g. increase in oxygen carrying capacity induced by hypoxia acclimatization) may play a significant role in some circumstances (e.g. in sedentary rats).

Acyl-CoA Dehydrogenase↗

PubMed1996

The supercooling ability of ticks (Acari, Ixodoidea).

The supercooling capacity of nine laboratory-held species of ticks originating from different geographical areas, as well as five field-collected species from Germany, was investigated. All but one tick species showed mean supercooling points between about -17 and -23 degrees C, suggesting that the capacity to supercool to temperatures of < or = -17 degrees C might be an inherent property of many tick species unrelated to their geographic origin. Photoperiod did not influence the mean supercooling point in any of the species and there was also no distinct seasonal pattern of supercooling in seasonally acclimatized Dermacentor marginatus. Thus, the supercooling ability was independent of the presence/absence of diapause. The finding of thermal hysteresis in D. marginatus hemolymph raises the question of whether or not anti-freeze proteins are involved in the supercooling capacity of that species. An interspecies comparison revealed a weak negative correlation between relative water content and supercooling point of the ticks and an even weaker correlation between body mass or body water mass and the supercooling point. Since the ticks exhibited low supercooling points both before and shortly after feeding, the blood used as food should lack potent ice nucleators.

Animals↗

PubMed1996

Benefits of training at moderate altitude versus sea level training in amateur runners.

After more than 25 years of research of altitude training (AT) there is no consensus regarding either the training programme at altitude or the effects of AT on performance at sea level. Based on a review of the research work on AT, we investigated combined base training and interval training at moderate altitude and compared immediate and delayed effects on sea level performance with those following similar sea level training (SLT). The altitude group (AG, 10 male amateur runners) trained at 2315 m (natural altitude) and the sea level group (SLG, 12 male amateur runners) at 187 m. Both groups performed 7 days of base training (running on a trail) lasting between 60 and 90 min a day and 5 days of interval training (speed and hill runs) for between 10 and 45 min a day. Incremental exercise tests were performed 1 week before (t1), 3 days after (t2) and 16 days after (t3) the 12-day main training period. Within AG, exercise performance improved from t1 to t2 by 8% (P < 0.05) and from t2 to t3 by 8% (P < 0.05). Maximum oxygen uptake (VO2max) increased from t2 to t3 by 10% (P < 0.05). Within SLG exercise performance increased from t2 to t3 by 8% (P < 0.05). At t3, relative and absolute VO2max in AG were significantly higher in comparison with SLG (P = 0.005 and P = 0.046 respectively). The improved performance 3 days after AT may be explained in part by an increased oxygen uptake at submaximal exercise intensities without a change in VO2max. Further enhancement in performance 2 weeks after AT, however, seems to have been due to the clearly enhanced VO2max. Progressive cardiovascular adjustments might have contributed primarily to the time-dependent improvements observed after AT, possibly by an enhanced stroke volume overcompensating the reduced heart rates during submaximal exercise. In conclusion, our findings would suggest that training at a moderate natural altitude improves performance at sea level more than SLT. Combining base and interval training with regulation of intensity by training at constant heart rates during acclimatization at altitude would seem to be a successful training regimen for amateur runners. Most beneficial effects became apparent during the subsequent SLT around 2 weeks after return from altitude. Therefore, we are convinced that AT should be reconsidered as a potent tool for enhancing aerobic capacity, at least in non-elite athletes.

Adult↗

PubMed1989

Thermoregulatory responses of firemen to exercise in the heat.

Twelve volunteer (VF) and 12 professional firemen (PF) wearing only brief trunks exercised on an electrically-braked cycle ergometer at three-five exercise intensities. After 45 min of exercise at 75 W, the exercise intensity was elevated in steps of 25 W every 15 min until the subject was exhausted. Air temperature was regulated to equal skin temperature (36 degrees-38 degrees C) and relative humidity was regulated at 52%. The two groups of firemen were comparable in terms of body mass, age and maximum oxygen consumption. Their oxygen consumption, rectal and skin temperatures, sweating and heart rate were measured during the tests. Blood lactate concentration was measured before, during and after the test. The physiological strain was higher in VF as indicated by higher heat storage, heart rate, skin and rectal temperatures. Sweat rate tended to be lower in VF than PF. The results indicated a better adaptation of the professional compared to the volunteer firemen to work in the heat, although the degree of heat acclimatization was considered to be equally minimal in both groups.

Adult↗

PubMed1999

Atrial natriuretic peptide relation to plasma and heart zinc concentrations of Wistar rats exposed to cold and hot ambients.

Plasma atrial natriuretic peptide (ANP) and zinc levels, as well as heart tissue zinc concentrations were determined, in male Wistar rats after the exposure of 114 rats at low temperature (4 degrees C) and 95 rats at high temperature (35-36 degrees C) for 28 d. Plasma ANP was estimated by radioimmunoassay and Zn2+ concentrations by atomic absorption spectrometry. Values were compared to a control group exposed at 20-22 degrees C (76 rats). Plasma ANP and Zn2+ levels, as well as heart tissue Zn2+ concentrations of control rats did not show statistically significant variations during the study, whereas rats exposed to cold and hot ambients showed significant variations of the parameters. A significant increase of plasma ANP and plasma zinc and heart tissue Zn2+ concentrations developed during cold exposure, whereas a gradual decrease of plasma ANP and Zn2+ levels was revealed during hot adaptation. Results also indicate that plasma ANP and zinc levels are proportionally related, whereas there is an inverse relationship between plasma ANP levels and heart Zn2+ concentrations, in both cold and hot exposed rats. In conclusion, our results show that ANP in relation to zinc probably play an important role in cold and hot acclimatization of rats.

Adaptation, Physiological↗

PubMedDate not supplied

Cadmium, mercury, and lead effects on gill tissue of freshwater crayfish Procambarus clarkii (Girard).

Intermolt adult crayfish P. clarkii were used for this work. After acclimatization to laboratory conditions crayfish were exposed to sublethal concentrations of cadmium, mercury, and lead for 96 h. Gills of control and exposed crayfish were removed and ATPase activity and oxygen uptake rate were determined. Structural damage of gill filaments was also observed. Gill tissue respiration rates were measured for individual crayfish using a Gilson differential respirometer. Lead causes a decrease of gill oxygen uptake, but neither cadmium nor mercury seems to affect it at the concentrations employed. Although all metals studied alter gill filament structure, lead damage is the most apparent. In the same way, significant differences in gill ATPase activity owing to metal exposure were only observed in lead treated crayfish.

Animals↗

PubMed1997

Bioavailability of rumen bacterial selenium in mice using tissue uptake technique.

A tissue uptake experiment was conducted to determine the bioavailability of rumen bacterial Selenium (Se) in mice. The donor animal was wether fed a diet containing 0.2 mg Se/kg dietary dry matter (DM). Ruminal fluid was collected 2 h postprandially. Bacterial-rich precipitate was obtained by differential centrifugation of the ruminal fluids. This was later freeze-dried and mixed in the diet to be used in feeding the mice experiment. Thirty growing female mice with a body wt (mean +/- SD) of 21.4 +/- 0.74 g were housed in plastic cages (5 mice/cage) and allotted equally to three dietary treatments. Diet 1 and Diet 2 were formulated based on AIN-76, except that no Se supplementation in the form of selenite was made in the former. In Diet 3, rumen bacterial matter was 20% of the diet, which gave an equivalent of 0.1 mg Se/kg dietary DM. The other two diets, Diet 1 and Diet 2, had an Se content of 0.025 and 0.1 mg/kg dietary DM, respectively. A 7-d feeding commenced after 7 d of acclimatization of the semipurified diet. Results showed that those mice fed an Se- (selenite) supplemented diet (Diet 2) had higher (P < 0.05) tissue Se concentrations than those mice fed the other two diets. No statistical differences were observed on various tissue Se concentrations between Diet 1 and Diet 3, although the latter diet had higher values. Kidney and liver had the highest Se concentrations compared to the other tissues. This study concludes that bacterial Se collected from the rumen of wether is not fully available for absorption in the intestine of the mice.

Animals↗

PubMed2000

Effect of altitude training on serum creatine kinase activity and serum cortisol concentration in triathletes.

In this investigation we evaluated the effect of a 5-week training program at 1860 m on serum creatine kinase (CK) activity and serum cortisol concentration in national-caliber triathletes for the purpose of monitoring the response to training in a hypobaric hypoxic environment. Subjects included 16 junior-level female (n = 8) and male (n = 8) triathletes who were training for the International Triathlon Union (ITU) World Championships. After an initial acclimatization period, training intensity and/or volume were increased progressively during the 5-week altitude training camp. Resting venous blood samples were drawn at 0700 hours following a 12-h overnight fast and were analyzed for serum CK activity and serum cortisol concentration. Subjects were evaluated before [7-10 days pre-altitude (SL 1)] and after [7-10 days post-altitude (SL 2)] the 5-week training camp at 1860 m. At altitude, subjects were evaluated within 24-36 h after arrival (ALT 1), 7 days after arrival (ALT 2), 18 days after arrival (ALT 3), and 24-36 h prior to leaving the altitude training camp (ALT 4). A repeated-measures analysis of variance was used to evaluate differences over time from SL 1 to SL 2. Compared to SL 1, serum CK activity increased approximately threefold (P < 0.05) within the initial 24-36 h at altitude (ALT 1), and increased by an additional 70% (P < 0.05) after the 1st week of altitude training (ALT 2). Serum CK activity remained significantly elevated over the duration of the experimental period compared to pre-altitude baseline levels. Serum cortisol concentration was increased (P < 0.05) at the end of the 5-week altitude training period (ALT 4) relative to SL 1, ALT 1 and ALT 3. These data suggest that: (1) the initial increase in serum CK activity observed in the first 24-36 h at altitude was due primarily to acute altitude exposure and was independent of increased training intensity and/or training volume, (2) the subsequent increases in serum CK activity observed over the duration of the 5-week altitude camp were probably due to the combined effects of altitude exposure and increased training load, and (3) the increase in serum cortisol concentration observed at the end of the altitude training camp reflects the additive effect of 5 weeks of altitude exposure in combination with a progressively increased training intensity and/or volume.

Adolescent↗

PubMed2001

Effect of endosulfan on antioxidants of freshwater fish Channa punctatus Bloch: 1. Protection against lipid peroxidation in liver by copper preexposure.

Effect of a single exposure of endosulfan (5 ppb) on catalase (CAT), glutathione peroxidase (GPx), glutathione S-transferase (GST), and reduced glutathione (GSH) of liver, kidney, and gill of a freshwater fish (Channa punctatus Bloch) were evaluated after 24 h of treatment. Endosulfan exposure resulted in a significant induction (p < 0.05-0.001) of GPx, GST activity, and GSH levels in all the organs. However, CAT activity was found to be significantly decreased (p < 0.01-0.001). Lipid peroxidation (LPO) values were also determined in liver, kidney, and gill and a significant increase in LPO values (p < 0.05-0.01) was observed in all the organs. We also investigated whether preexposure to low concentration of copper (10 ppb) for 4 weeks has any protective effect against endosulfan-induced oxidative damage. In copper-acclimatized endosulfan-exposed fish, a significant decrease in GPx (p < 0.001), GST (p < 0.05), GSH (p < 0.001) levels, and LPO (p < 0.01) was observed in the liver, whereas CAT activity was increased significantly (p < 0.001). However, kidney and gill did not show any significant alterations in antioxidant levels. The results of this study demonstrate that endosulfan induces peroxidative damage in liver, kidney, and gill in response to which levels of antioxidant were modulated. However, when fish preacclimatized to copper were exposed to endosulfan, protection against oxidative damage was observed only in the liver. It is proposed that measurement of antioxidants in fish tissues may prove to be useful in biomonitoring of exposure to aquatic pollutants.

Animals↗

PubMed1997

Effects of organophosphate and carbamate pesticides on acetylcholinesterase and choline acetyltransferase activities of the polychaete Nereis diversicolor.

A toxicity test for organophosphates (OP) and carbamates (C) was improved with the adult ragworm Nereis diversicolor. Animals were maintained in U-shaped glass tubes of 4-mm inner diameter fixed vertically on a plastic plate and placed in glass aquaria. Each tank was covered with glass in order to reduce evaporation and heat dissipation. Temperature varied between 15 and 16 degrees C and salinity was constant (34 per thousand) during the entire length of the experiment. Experiments were performed with a fixed day length of 12 h and seawater was gently aerated. The maintenance system allowed the administration of OP and C compounds via the seawater. An acclimatization period of 48 h was not sufficient to accomodate worms to their artificial burrows; accordingly, we chose to acclimate worms for a week before beginning the exposure. Choline acetyltransferase (ChAT) activity was very low and was not significantly modified by two OP compounds: malathion and parathion-ethyl. ChAT is not a target for these pesticides and should not be used for future studies about OP and C toxicity. On the other hand, inhibitory effects on acetylcholinesterase (AChE) activity were determined at concentrations of 10(-6) M for three OP compounds-malathion, parathion-ethyl, and phosalone-and a carbamate pesticide-carbaryl. We measured only short-term effects and no cumulative effect was determined, the maximum percentage of AChE activity inhibition being between 2 (carbaryl) and 7 (OP compounds) days after exposure and then remaining stable. Mortality occured only after a period of intoxication of 14 days. N diversicolor, which can be easily maintained at the laboratory, seems to be a good candidate for future laboratory studies to test the toxicity of other pollutants.

Animals↗

PubMed1997

Comparative fate of 1,1-diphenylethylene (DPE), 1,1-dichloro-2,2-bis(4-Chlorophenyl)-ethylene (DDE), and pentachlorophenol (PCP) under alternating aerobic and anaerobic conditions.

Bacterial degradation of 1,1-dichloro-2,2-bis-(4-chlorophenyl)-ethylene (DDE) and its dehalogenated derivative 1,1-diphenylethylene (DPE) has not yet been shown and may require culture adaptation and special culture conditions. We compared the degradability of DPE, DDE, and pentachlorophenol (PCP) in aerobic/anaerobic sequenced batch reactor systems. Reactors operated under aerobic/methanogenic and aerobic/denitrifying conditions were inoculated with bacterial consortia from anaerobic granular sludge, long-term PCP- and DDE-contaminated soil, and pulp and paper waste pond sediment. The culture was gradually acclimatized to low concentrations of DPE, DDE, and PCP in defined minimal growth media with benzoate, phenol, ethanol, and formate as primary carbon sources. DDE remained refractory for 105 days, whereas DPE and PCP were degraded. This suggests that DDE is extremely recalcitrant to degradation by aromatic organochlorine-degrading bacteria from long-term polluted soils and sediments. The results confirm that the chlorination of DDE is a major biodegradation barrier for adapted bacteria under aerobic and anaerobic conditions.

Bacteria, Aerobic↗

PubMed2002

Effects of nitrite and ammonium on methane-dependent denitrification.

For effective application of methane-dependent denitrification (MDD) in the treatment of wastewater containing NO(2)(-) or NH(4)(+), the effect of these inorganic nitrogen compounds on MDD activity needs to be clarified. The MDD activity of sludge acclimatized with CH(4) and O(2) was determined with mineral media of different nitrogen-compound compositions in the presence of 0.21 atm CH(4) and 0.20 atm O(2). Incubations with media containing only NO(2)(-) or two of the three inorganic nitrogen compounds (NO(3)(-)+NO(2)(-), NO(2)(-)+NH(4)(+) or NH(4)(+)+NO(3)(-)) resulted in MDD activity equal to or higher than that with media containing only NO(3)(-). However, there was no MDD activity in media containing NO(2)(-) at 10 degrees C, probably because of serious inhibition of NO(2)(-) on methane oxidation. MDD occurred in media containing only NH(4)(+), although the total nitrogen removal efficiency was very low. These results show that NO(2)(-) and NH(4)(+), in the presence of NO(x)(-), do not inhibit but rather promote MDD. Consequently, NH(4)(+) does not need to be completely oxidized to NO(3)(-) in the nitrification reactor before MDD. However, under psychrophilic conditions, NO(2)(-) seriously inhibited MDD. Therefore, the nitrification reactor must not discharge effluent containing NO(2)(-) under psychrophilic conditions.

Methane↗

PubMed2006

Influence of temperature on process efficiency and microbial community response during the biological removal of chlorophenols in a packed-bed bioreactor.

Two reactors, initially operated at 14 and 23+/-1 degrees C (RA and RB, respectively), were inoculated with a bacterial consortium enriched and acclimatized to the respective temperatures over 4 months. The biofilms, formed in the reactors, were studied using scanning electron microscopy, cultivation of the biofilm microflora, and physiological analysis of the isolates. Two bacteria able to mineralize chlorophenols under a large range of temperature (10-30 degrees C) were isolated from the biofilm communities of reactors RA and RB and characterized as Alcaligenaceae bacterium R14C4 and Cupriavidus basilensis R25C6, respectively. When temperature was decreased by 10 degrees C, the chlorophenols removal capacity was reduced from 51.6 to 22.8 mg l(-1) h(-1) in bioreactor RA (from 14 to 4 degrees C) and from 59.3 to 34.7 mg l(-1) h(-1) in bioreactor RB (from 23+/-1 to 14 degrees C). Fluorescence in situ hybridization (FISH) of the biofilm communities showed that, in all temperatures tested, the beta-proteobacteria were the major bacterial community (35-47%) followed by the gamma-proteobacteria (12.0-6.5%). When the temperature was decreased by 10 degrees C, the proportions of gamma-proteobacteria and Pseudomonas species increased significantly in both microbial communities.

Alcaligenaceae↗

PubMed2004

Effect of different temperature downshifts on protein synthesis by Aeromonas hydrophila.

The psychrotrophic bacterium Aeromonas hydrophila 7966 was subjected to cold shocks from 30 degrees C to 20 degrees C, 15 degrees C, 10 degrees C, or 5 degrees C, or were incubated at low temperature to determine its adaptative response. The cell protein patterns analyzed by two-dimensional electrophoresis revealed that only a few proteins were underexpressed, whereas numerous new proteins appeared with the decrease of temperature, and some others were overexpressed. Among them, a few constituted cold shock proteins because they were transiently induced, whereas others belong to the acclimatation family proteins. Two cold shock proteins of 11 kDa were synthesized at low level because they were visualized only after radiolabeling or silver staining. Moreover, under our experimental conditions, no major cold shock protein of a molecular mass similar to that of E. coli (7.4 kDa) could be identified.

Adaptation, Physiological↗

PubMed2003

Agrobacterium tumefaciens-mediated transformation of Indian mulberry, Morus indica cv. K2: a time-phased screening strategy.

An efficient and reproducible protocol for the production of transgenic plants was developed for Morus indica cv. K2 by Agrobacterium tumefaciens-mediated transformation. The hypocotyls, cotyledon, leaf and leaf callus explants precultured for 5 days on regeneration medium were co-cultivated with a bacterial suspension at 10(9) cells/ml for 3 days in the dark. Infectivity of A. tumefaciens strain LBA4404 was more than that of strains GV2260 and A281, and among the various plasmids tried, pBI121 and pBI101:Act1 transformed nearly 100% of the explants followed closely by p35SGUSINT. About 90-100% of the explants tested positive in the beta-glucuronidase (GUS) histochemical assay performed after 3 days of co-cultivation. This high level of transient expression, however, decreased to 20-25% after 15 days. Gus activity was most stable in the callus explants, which emerged as the explant of choice for transformation. The transformed explants were selected on 50-75 mg/l kanamycin for 1 month, and 25-50% of the explants developed adventitious buds. On the basis of kanamycin-resistant shoots produced from the total number of explants inoculated, the transformation efficiency was 44%. After 1 month, 40% of these shoots displayed high gus activity as assessed by the GUS fluorometric assay. On a selection-free root induction medium, 80% of the shoots developed roots and 90% of the potted plantlets acclimatized to the growth room conditions. The 3-month-old regenerates showed gus and nptII(neomycin phosphotransferase II) gene activity as assayed by the GUS fluorometric assay and nptII enzyme assay, followed by PCR polymerase chain reaction (54.5%) analysis after 6-months. Transgene integration into the nuclear genome of 1-year-old regenerates was confirmed in 10 of the 18 transformants tested by Southern analysis. The transformation efficiency as defined by the number of transgenic plants produced from the total number of explants co-cultivated was 6%.

Agrobacterium tumefaciens↗