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71.

Background

Safflower (Carthamus tinctorius L.) is a difficult crop to genetically transform being susceptible to hyperhydration and poor in vitro root formation. In addition to traditional uses safflower has recently emerged as a broadacre platform for the production of transgenic products including modified oils and pharmaceutically active proteins. Despite commercial activities based on the genetic modification of safflower, there is no method available in the public domain describing the transformation of safflower that generates transformed T1 progeny.

Results

An efficient and reproducible protocol has been developed with a transformation efficiency of 4.8% and 3.1% for S-317 (high oleic acid content) and WT (high linoleic acid content) genotypes respectively. An improved safflower transformation T-DNA vector was developed, including a secreted GFP to allow non-destructive assessment of transgenic shoots. Hyperhydration and necrosis of Agrobacterium-infected cotyledons was effectively controlled by using iota-carrageenan, L-cysteine and ascorbic acid. To overcome poor in vitro root formation for the first time a grafting method was developed for safflower in which ~50% of transgenic shoots develop into mature plants bearing viable transgenic T1 seed. The integration and expression of secreted GFP and hygromycin genes were confirmed by PCR, Southern and Western blot analysis. Southern blot analysis in nine independent lines indicated that 1-7 transgenes were inserted per line and T1 progeny displayed Mendelian inheritance.

Conclusions

This protocol demonstrates significant improvements in both the efficiency and ease of use over existing safflower transformation protocols. This is the first complete method of genetic transformation of safflower that generates stably-transformed plants and progeny, allowing this crop to benefit from modern molecular applications.
  相似文献   
72.
Rates of degradation of seven organophosphate nematicides and insecticides were examined in two soils known to show accelerated biodegradation of fenamiphos and one soil known to show accelerated biodegradation of chlorpyrifos. The results indicated that several organophosphate insecticides and one nematicide were susceptible to cross-enhanced degradation in the soil showing accelerated biodegradation of chlorpyrifos. No cross-enhancement was observed in the two soils showing accelerated degradation of fenamiphos. Fumigation resulted in the complete inhibition of pesticide degradation in all soils. The data suggested that the cross-enhancement of selected pesticides in chlorpyrifos-degrading soil was dependent on the structural similarity of the compounds. Mechanisms of degradation of pesticide in soil support this hypothesis, where structurally similar compounds (diazinon, parathion, coumaphos and isazofos) were hydrolysed by microbial activity in chlorpyrifos-degrading soil but the degradation products were accumulated. Enhanced degradation of chlorpyrifos and fenamiphos was found to be stable in the laboratory condition for a period of one year.  相似文献   
73.
The objective of this study was to establish a withdrawal period for flunixin in milk by quantifying 5-hydroxyflunixin, the marker residue, in bovine milk as a function of time, following intravenous treatment of lactating dairy cows with flunixin-N-methyl glucamine (Banamine or Finadyne). Lactating dairy cows were dosed on three consecutive days at 2.2 mg of flunixin free acid/kg of body weight/day. Milk was collected twice daily and assayed using a liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) procedure. The method was validated at concentrations in the range 0.5-250 ppb. The concentrations for 5-hydroxyflunixin measured 12 h after the last administration of drug ranged from 1.56 to 40.6 ppb for all cows. Milk concentrations for 5-hydroxyflunixin were used to establish withdrawal periods of 36 h using guidelines established by the U.S. Food and Drug Administration/Center for Veterinary Medicine and 24 h using guidelines established by the European Medicinal Evaluation Agency/Committee on Veterinary Medicinal Products.  相似文献   
74.
Disease of geese caused by a new herpesvirus   总被引:3,自引:0,他引:3  
A goose flock farmed outdoors in south-eastern Queensland suffered an outbreak of peracute disease with high death rate (97%). Small button ulcers and large plaques overlying lymphocyte aggregates were present on the mucosa of the small intestine of affected birds. Small white foci of necrosis and focal haemorrhages were seen in the livers. Numerous intranuclear inclusion bodies were observed microscopically in hepatocytes and a herpesvirus which grew rapidly in chicken kidney cells was isolated from tissues. Duck virus enteritis (DVE) was suspected but DVE antiserums failed to neutralise the virus. Further serological studies with a limited range of known avian herpesviruses have failed to identify the virus. Experimental transmission resulted in high mortality in geese (100%), lower mortality in ducklings and nil mortality in chickens. Surveillance studies showed no evidence of infection in domestic and wild birds beyond the original farm and the infection appears not to have been established in the area. Wild ducks, which were frequent visitors to the farm dam, were considered the most likely source of the infection.  相似文献   
75.
Total uptake of linuron by wheat seedlings in nutrient solutions was close to that expected from the product of the amounts of water transpired by the plants and the concentrations of herbicide in solution. Uptake from 19 different soils was less than the amount supplied by mass-flow when the concentrations of linuron in the soil solution were estimated from slurry adsorption measurements. Using a pressure-membrane technique, it was shown that the actual soil solution concentrations of linuron were less than those estimated, and following rewetting of pressure-membrane samples, the rate of redistribution of linuron between the adsorbed and solution phases was slow. The results suggest that under the conditions of the uptake experiments, the systems were not in equilibrium, and show that the rates of adsorption and desorption of linuron may be important in determining its availability to plants.  相似文献   
76.
The persistence of the herbicide 2,4,5-T was studied at different controlled temperatures and moisture levels in Regina heavy clay. Degradation approximated to first-order kinetics and the half-life varied from about 4 days at 35°C and 34% soil moisture to about 60 days at 10°C and 20% soil moisture. The laboratory data were used in conjunction with the appropriate measurements of surface soil temperature and moisture content in the field to simulate the degradation pattern for the herbicide in five separate micro-plot experiments. Satisfactory agreement with the observed patterns of loss was obtained in two of the experiments but in the other three, the model over-estimated rates of loss. It is suggested that the reason for this was the difficulty of obtaining a correct measure of soil moisture content to use in the simulation program.  相似文献   
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