Growth Response of Wheat (<i>Triticum aestivum</i>) Callus to Imazapyr and in Vitro Selection for Resistance

Heering, David C.; Guenzi, Arron C.; Peeper, Thomas F.; Claypool, P. L.

doi:10.1017/s0043174500057179

Cited by 5 publications

(3 citation statements)

References 19 publications

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“…The concentrations of imazapyr, imazethapyr, or imazaquin as high as 500 μg/l scarcely affected sugarcane callus biomass and embryogenicity and also did not result in cell death. This is similar to the result found by Heering et al (1992), where wheat calli were less sensitive to 100 μM imazapyr, in contrast to the whole wheat plant. Therefore, the effectiveness of herbicide concentrations on growth differs significantly across plant tissues and plant species.…”

Section: Discussionsupporting

confidence: 92%

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In vitro selection of transgenic sugarcane callus utilizing a plant gene encoding a mutant form of acetolactate synthase

Vyver

Conradie

Koßmann

et al. 2013

In Vitro Cell.Dev.Biol.-Plant

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Selection genes are routinely used in plant genetic transformation protocols to ensure the survival of transformed cells by limiting the regeneration of non-transgenic cells. In order to find alternatives to the use of antibiotics as selection agents, we followed a targeted approach utilizing a plant gene, encoding a mutant form of the enzyme acetolactate synthase, to convey resistance to herbicides. The sensitivity of sugarcane callus (Saccharum spp. hybrids, cv. NCo310) to a number of herbicides from the sulfonylurea and imidazolinone classes was tested. Callus growth was most affected by sulfonylurea herbicides, particularly 3.6 μg/l chlorsulfuron. Herbicide-resistant transgenic sugarcane plants containing mutant forms of a tobacco acetolactate synthase (als) gene were obtained following biolistic transformation. Post-bombardment, putative transgenic callus was selectively proliferated on MS medium containing 3 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D), 20 g/l sucrose, 0.5 g/l casein, and 3.6 μg/l chlorsulfuron. Plant regeneration and rooting was done on MS medium lacking 2,4-D under similar selection conditions. Thirty vigorously growing putative transgenic plants were successfully ex vitro-acclimatized and established under glasshouse conditions. Glasshouse spraying of putative transgenic plants with 100 mg/l chlorsulfuron dramatically decreased the amount of non-transgenic plants that had escaped the in vitro selection regime. PCR analysis showed that six surviving plants were als-positive and that five of these expressed the mutant als gene. This report is the first to describe a selection system for sugarcane transformation that uses a selectable marker gene of plant origin targeted by a sulfonylurea herbicide.

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Section: Discussionsupporting

confidence: 92%

“…A similar result has been reported for the mannose selection system in sugarcane where different concentrations of mannose inhibited growth by 50–80%, but could not completely suppress callus growth (Jain et al 2007). Also, wheat callus exposed to imazapyr only showed 50% growth inhibition after 70 d. (Heering et al 1992). …”

Section: Discussionmentioning

confidence: 99%

In vitro selection of transgenic sugarcane callus utilizing a plant gene encoding a mutant form of acetolactate synthase

Vyver

Conradie

Koßmann

et al. 2013

In Vitro Cell.Dev.Biol.-Plant

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“…Crops resistant to ALS inhibitors have the potential to overcome carryover injury and possibly provide selectivity, allowing the use of desirable herbicides directly in resistant crops. Crops resistant to ALS-inhibiting herbicides have been derived by somatic cell selection (Anderson and Georgeson 1989;Heering et al 1992;Iler et al 1993;Saunders et al 1992), seed mutagensis (Newhouse et all 1992;Sebastian et al 1989), plant transformation (D'Halluin et al 1992;Haughn et al 1988;McHughen 1989), and interspecific crosses (Mallory-Smith et all 1990).…”

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confidence: 99%

Biochemical mechanism and molecular basis for ALS-inhibiting herbicide resistance in sugarbeet (Beta vulgaris) somatic cell selections

Wright

Bascomb²,

Sturner³

et al. 1998

Weed sci.

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Three sugarbeet selections differing in cross-resistance to three classes of acetolactate synthase (ALS)-inhibiting herbicides have been developed using somatic cell selection. Sugarbeet selections resistant to imidazolinone herbicides,Sir-13and93R30B, do not metabolize [14C]-imazethapyr any faster or differently than sensitive, wild-type sugarbeets or a sulfonylurea-resistant/imidazolinone-sensitive selection, Sur. ALS specific activity from the three herbicide-resistant selections ranged from 73 to 93% of the wild-type enzyme extracts in the absence of herbicide, indicating enzyme overexpression was not a factor in resistance. Acetolactate synthase from Sir-13 plants showed a 40-fold resistance to imazethapyr but no resistance to chlorsulfuron or flumetsulam. Polymerase chain reaction amplification and sequencing of two regions of the ALS gene spanning all known sites for ALS-based herbicide resistance in plants indicated a single nucleotide change in theSir-13gene (G337to A337) resulting in a deduced substitution of threonine for alanine at position 113 in the sugarbeet amino acid sequence. Sur ALS was not significantly resistant to imazethapyr, but was 1,000- and 50-fold resistant to chlorsulfuron and flumetsulam, respectively.Surgene sequencing indicated a single nucleotide change(C562to T562) resulting in a serine for proline substitution at position 188 of the ALS primary structure. The93R30Bnucleotide sequence indicated two mutations resulting in two deduced amino acid substitutions: threonine for alanine at position 113 plus serine for proline at position 188. The93R30Bdouble mutant incorporated the changes observed in each of the single mutants above and correlated with higher resistance levels to imazethapyr (> 1,000-fold), chlorsulfuron (4,300-fold), and flumetsulam (200-fold) at the ALS level than observed in either of the single mutants.93R30Brepresents the first double mutant derived by a two-step selection process that incorporates two class-specific ALS-inhibitor resistance mutations to form a single broad cross-resistance trait. The interaction of the two altered amino acids is synergistic with respect to enzyme resistance vs. the resistance afforded by each of the individual mutations.

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Factors affecting the establishment and maintenance of embryogenic callus and suspension cultures of wheat (Triticum aestivum L.)

1995

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Improved suspension cell culture systems are needed to facilitate the application of recombinant DNA technology for wheat germplasm enhancement. This study evaluated three wheat (Triticum aestivum L.) cultivars, and the effects of medium basal salts, 2,4-D, sucrose, and L-proline concentrations on the establishment of rapidly growing and highly embryogenic callus and suspension cultures. Percent embryogenic calli was visually estimated and verified with light and scanning electron microscopy. The most highly embryogenic callus was produced by cultivar Bobwhite on medium with MS basal salts, 5.6 μ M 2,4-D, 58 mM sucrose, and zero proline. The suspension cultures that produced the greatest number of regenerated plants utilized callus tissue produced on solid medium with MS basal salts, 87 mM sucrose, 9 μM 2,4-D, and no proline.

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Growth Response of Wheat (Triticum aestivum) Callus to Imazapyr and in Vitro Selection for Resistance

Cited by 5 publications

References 19 publications

In vitro selection of transgenic sugarcane callus utilizing a plant gene encoding a mutant form of acetolactate synthase

In vitro selection of transgenic sugarcane callus utilizing a plant gene encoding a mutant form of acetolactate synthase

Biochemical mechanism and molecular basis for ALS-inhibiting herbicide resistance in sugarbeet (Beta vulgaris) somatic cell selections

Factors affecting the establishment and maintenance of embryogenic callus and suspension cultures of wheat (Triticum aestivum L.)

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