Construction of plant transformation vectors carrying beet necrotic yellow vein virus coat protein gene (ii)- plant transformation

Fragments containing the coat protein gene of beet necrotic yellow vein virus were cloned in two plant transformation vectors: pCAMBIA3301M with the bar gene as selectable marker, and pCAMBIA1304M, with resistance to hygromycin. Three constructs were made of each vector: CPL, containing coat protein gene with leader sequence; CPS with coat protein gene, and CPSas with coat protein gene in antisense orientation. Vectors pC3301MCPL, pC3301MCPS. and pC3301MCPSas were used in Agrobacterium—mediated transformation of Nicotiana tabacum (tobacco), Nicotiana excelsior and Nicotiana benthamiana. Regenerants that developed roots on selective media were tested for the presence of CP fragments and the bar gene, but most regenerants were nontransformed (50-83% escapes). After all rooted plants had been selfed, and T1 seed germinated on selective media, only plants descending from one N. excelsior regenerant transformed with pC3301MCPS were positive for presence of bar gene and CPS fragment. Tobacco and Nicotiana benthamiana were transformed with constructs pC1304MCPS and pC1304MCPSas. Transformation efficiency was much higher and approximately 50% of regenerants that rooted on media with 20 mg l−1 hygromycin were positive for the presence of CP fragments. All T1 plants were positive for presence of CP fragments

Teeltonderzoek : biologische suikerbietenteelt

Verslag van de resultaten in het jaar 2002 in het onderzoek naar biologische suikerbietenteelt. Er wordt kort omschreven welke ziekten en plagen een risico opleveren als suikerbieten biologisch worden geteeld

Metabolic studies on Asulam (M&B 9057).

The well-established herbicide, asulam (methy14-aminobenzenesulphonylcarbamate; M&B 9057) is used widely for control of weed species including docks and bracken. The compound resembles sulphonamides in structure. Most of an oral dose of asulam is eliminated from rats mainly in the urine (75%) and faeces (19%) within 24 hours. Biotransformation in the intact animal is similar to the N-acetylation of some sulphonamides. The major metabolite is N4-acetylasulam which is excreted rapidly in the urine (15% male; 9% female) and also appears in faeces (4%). Induction of rats with asulam does not effect overall metabolism and excretion except that decarbamoylation is increased in the gut. N4-acetylsulphanilamide is also produced ( 2%) by decarbamoylation in the gut. N4-acetylasulam formation in vitro is located in the cell-membrane fraction of liver. Asulam is not eliminated in the bile (1% dose) in vivo. No binding spectrum is obtained with microsomal cytochrome P450. Continuous perfusion through liver for 6 hours does not increase biliary elimination, but at least three water-soluble unidentified compounds (4.5%) are formed in plasma. In contrast to the in vivo state asulam causes rapid oxygen uptake in vitro with microsomes producing an isolatable colored metabolite (max = 330 nm) which contains ring-substituted hydroxy sulphonamido and nitrogen substituted carbamate groups. Mass spectrometry shows this not to be 2-hydroxyasulam

Metabolic studies on Asulam (M&B 9057).

The well-established herbicide, asulam (methy14-aminobenzenesulphonylcarbamate; M&B 9057) is used widely for control of weed species including docks and bracken. The compound resembles sulphonamides in structure. Most of an oral dose of asulam is eliminated from rats mainly in the urine (75%) and faeces (19%) within 24 hours. Biotransformation in the intact animal is similar to the N-acetylation of some sulphonamides. The major metabolite is N4-acetylasulam which is excreted rapidly in the urine (15% male; 9% female) and also appears in faeces (4%). Induction of rats with asulam does not effect overall metabolism and excretion except that decarbamoylation is increased in the gut. N4-acetylsulphanilamide is also produced ( 2%) by decarbamoylation in the gut. N4-acetylasulam formation in vitro is located in the cell-membrane fraction of liver. Asulam is not eliminated in the bile (1% dose) in vivo. No binding spectrum is obtained with microsomal cytochrome P450. Continuous perfusion through liver for 6 hours does not increase biliary elimination, but at least three water-soluble unidentified compounds (4.5%) are formed in plasma. In contrast to the in vivo state asulam causes rapid oxygen uptake in vitro with microsomes producing an isolatable colored metabolite (max = 330 nm) which contains ring-substituted hydroxy sulphonamido and nitrogen substituted carbamate groups. Mass spectrometry shows this not to be 2-hydroxyasulam