Transformation with a mutant Arabidopsis acetolactate synthase gene renders tobacco resistant to sulfonylurea herbicides

Summary A gene encoding acetolactate synthase was cloned from a chlorsulfuron-resistant mutant of Arabidopsis. The DNA sequence of the mutant gene differed from that of the wild type by a single base pair substitution. When introduced into tobacco by Ti plasmid-mediated transformation the gene conferred a high level of herbicide resistance. These results suggest that the cloned gene may confer agronomically useful levels of herbicide resistnace in other crop species, and that it may be useful as a selectable marker for plant transformation experiments.     

Chlorsulfuron resistance in Daucus carota cell lines and plants:Involvement of gene amplification

Daucus carota L. cell lines stably resistant to the herbicide chlorsulfuron (CS) have been isolated according to a stepwise selection. Studies carried out during different selection steps show that the specific activity of the target enzyme acetohydroxyacid synthase (AHAS) increases along with CS resistance. Southern hybridization analysis performed with aBrassica napus AHAS probe in a CS highly-resistant cell line reveals the presence of a greatly amplifiedEcoRI fragment of genomic DNA. This indicates that AHAS overproduction induced by stepwise selection is due to gene amplification. Regenerants from some resistant cell lines maintained the CS-resistant trait at the whole plant level.     

Rhizoremediation of residual sulfonylurea herbicides in agricultural soils using Lens culinaris and a commercial supplement

Sulfonylureas (SU) are a popular herbicide used today for controlling weeds. While beneficial for this purpose they present a persistent problem in agricultural treated areas, with this treatment proving detrimental for successive crops. This study assessed the phytoremediative properties of lentils (Lens culinaris) grown in uncontaminated and chlorsulfuron-contaminated soil, with and without the addition of a growth supplement, PulseAider?. The results show that in the presence of lentils the degradation of chlorsulfuron is enhanced and this degradation rate is significantly increased when the PulseAider? supplement was included during seed sowing. The supplement PulseAider? also significantly increased shoot and root biomass, root branching, and nodule number under control conditions. While this was not so for plants grown in contaminated soils, the PulseAider? supplement seemed to alter root branching and morphology. Most Probable Number (MPN) assays showed increased numbers of potential chlorsulfuron-degrading bacteria in soil treated with PulseAider?, although this was found to be significant only in the control soil. Sequencing of the 16S ribosomal gene showed the presence of Pseudomonas fluorescens bacterial species which is a known chlorsulfuron-degrading bacterium. This study is one of the first to address the remediation of residual SU herbicides and offers an economically feasible solution that may have an impact on global food security.     

Chlorsulfuron influence on garden pea reproduction

The influence of chlorsulfuron on the reproduction of green pea ( Pisum sativum ) was examined by exposing plants at 3 different stages of development to 3 different exposure levels (46, 92, and 180 mg ha⁻¹; corresponding to 2 × 10⁻³, 4 × 10⁻³, and 8 × 10⁻³ of the recommended field application rates for small grain crops). Reproduction was only influenced by the two higher rates. The most susceptible stage of development was when plants possessed 6 expanded leaves and one visible flower bud. At that stage, an application rate of 180 mg ha⁻¹ (0.8% of the recommended field rate) reduced the seed yield of treated plants to 1% of that of the control plants without severely altering the height or appearance of mature plants. When correspondingly low application rates of atrazine. glyphosate, and 2.4-D were administered at this same developmental stage there were no effects on either growth or reproduction. Thus, chlorsulfuron had an influence on plant reproduction that was not produced by other herbicides administered at low levels.     

The effect of spray volume on spray partitioning between plant and soil

Summary Applications of chlorsulfuron (11.25 g ai ha⁻¹) were made to wheat, flax, canola and lentils at spray volumes of 48, 108 and 2171 ha⁻¹, and with metsulfuron methyl (6.00 g ai ha⁻¹) at spray volumes of 48 and 2171 ha⁻¹. Applications were made to the shoot only, the soil only and to the plant plus soil. Spectrofluorometric analysis was used to determine spray partitioning within the plant-soil system and foliar retention was related to efficacy. Fresh and dry weights of shoot material were determined 3 weeks after treatment. Flax and wheat were more tolerant of restricted-foliar applications than those made to the soil, the converse being true to canola and lentils. Applications made to the plant and soil were always the most deleterious. Foliar retention and efficacy did not correlate directly. Applications in 2171 ha⁻¹ were generally more efficacious than those at 481 ha⁻¹.     

Inhibition of fusicoccin-stimulated K+/H+ transport in root tips from maize seedlings pretreated with Chlorsulfuron

Abstract Fusicoccin (FC)-stimulated K⁺ (₈₆Rb) uptake and proton extrusion of maize (Zea mays) root apical segments were inhibited by pretreatment of 4-day-old seedlings with the herbicide Chlorsulfuron. In the range of Chlorsulfuron concentrations 0.01-10 mmol m^?3, the percentage of inhibition was 15% at 0.01 mmol m^?3 and progressively increased with Chlorsulfuron concentration up to 60% at 10 mmol m^?3. At the maximum concentration tested (10 mmol m^?3), the inhibition was evident after 1.5 h of pre-treatment. The binding of FC to microsomal fractions of root segments from Chlorsulfuron-pretreated seedlings was inhibited by 30%. It is suggested that Chlorsulfuron causes an alteration at the plasmalemma level involving the FC binding sites. The ineffectiveness of Chlorsulfuron in inhibiting FC-stimulaled K⁺ uptake when administered to excised segments, while inhibiting the enzyme acetolactate synthase, pointed out by Ray (1984) as the site of action of Chlorsulfuron in pea plants, suggests that the observed inhibition of K⁺ uptake and H⁺ extrusion is not induced by Chlorsulfuron inhibition of this enzyme. An alternative site of action of Chlorsulfuron is hypothesized in maize plants.     

Herbicide-tolerant corn by pollen selection

Summary Maize pollen was exposed to the herbicide Chlorsulfuron (CS), and segregation for tolerance was observed. The resulting plant generation exhibited significantly greater tolerance to CS than other (control) progeny. Since the increase in tolerance occurred after only one generation of pollen exposure, this result demonstrates that pollen selection can be used to develop herbicide-resistant crop species, even when the species are not amenable to cell culture. It also suggests a possible mechanism for the rapid evolution of herbicide tolerance in weeds.     

Agrobacterium mediated transfer of a mutant Arabidopsis acetolactate synthase gene confers resistance to chlorsulfuron in chicory (Cichorium intybus L.)

Summary Leaf discs of C. intybus were inoculated with an Agrobacterium tumefaciens strain harboring a neomycin phosphotransferase (neo) gene for kanamycin resistance and a mutant acetolactate synthase gene (csr1-1) from Arabidopsis thaliana conferring resistance to sulfonylurea herbicides. A regeneration medium was optimized which permitted an efficient shoot regeneration from leaf discs. Transgenic shoots were selected on rooting medium containing 100 mg/l kanamycin sulfate. Integration of the csr1-1 gene into genomic DNA of kanamycin resistant chicory plants was confirmed by Southern blot hybridizations. Analysis of the selfed progenies (S1 and S2) of two independent transformed clones showed that kanamycin and chlorsulfuron resistances were inherited as dominant Mendelian traits. The method described here for producing transformed plants will allow new opportunities for chicory breeding.     

Cell selection and inheritance of imidazolinone resistance in sugarbeet (Beta vulgaris)

 Sugarbeets are sensitive to imidazolinone herbicide residues applied to rotational crops. Two imidazolinone-resistance (IMI-R) sugarbeet traits were developed by somatic cell selection to overcome rotation restrictions for sugarbeets where imidazolinones have been applied. Sir-13 is an IMI-R/SU-S (sulfonylurea-sensitive) variant selected from an imidazolinone-sensitive (IMI-S) sugarbeet clone, REL-1. A second variant, 93R30B, resistant to imidazolinone as well as to sulfonylurea herbicides (IMI-R/SU-R), was selected from a plant homozygous for a previously described sulfonylurea-specific resistance trait, Sur (IMI-S/SU-R). The IMI-R alleles (Sir-13 and 93R30B) were found to be corresponding allelic variants at the same ALS locus and both were tightly associated with the Sur allele. Each resistant allele is dominant to the sensitive wild-type allele; however, incomplete dominance is shown among resistance alleles. Diploid sugarbeet contains a single ALS gene copy, limiting the ability to stack these resistance traits in the same plant by traditional breeding. Received: 1 May 1997 / Accepted: 30 June 1997