Plant cell wall extensibility is mediated, in part, by xyloglucan endotransglycosylases/hydrolases (XTH) that are able to cleave and reattach xyloglucan polymers that make up the hemicelluloses matrix of type I cell walls. In Arabidopsis and other plants, XTHs are encoded by relatively large gene families that are regulated in specific spatial and temporal patterns. In silico screening of a cotton expressed sequence tag (EST) database identified 23 sequences with close sequence similarity to Arabidopsis XTH coding sequences. Analysis of full-length cotton cDNAs derived from these ESTs allow for the identification of three distinct GhXTH cDNAs (denoted GhXTH1, GhXTH2 and GhXTH3) based primarily on their 3' untranslated sequences. The three GhXTH genes were expressed differently with GhXTH1 predominantly expressed in elongating cotton fibers. The function of GhXTH1 in mediating cotton fiber elongation was analyzed in transgenic cotton plants that express a transgene consisting of the GhXTH1 coding sequence under transcriptional control of the CaMV 35S promoter. Plants that over-expressed GhXTH1 had increased XTH activity and produced mature cotton fibers that were between 15 and 20% longer than wild-type cotton plants under both greenhouse and field growth conditions. Segregation analysis showed that the 35S::GhXTH1 transgene acts as a dominant fiber length allele in transgenic cotton. These results confirm that GhXTH1 is the predominant XTH in elongating fibers and its expression limits cotton fiber elongation.
Variability in weed control following pyrithiobac applications has been observed under field conditions. The influence of temperature on this variability was investigated. Results from field studies performed over two growing seasons identified plant and air temperatures at the time of herbicide treatment that correlated with whole-plant efficacy differences. Based on the field data, weed control with pyrithiobac was acceptable at application temperatures of 20 to 34 C. To investigate a potential source of thermal limitations on pyrithiobac efficacy, the thermal dependence of in vitro inhibition of acetolactate synthase (ALS), the site of action for pyrithiobac, was examined. A crude leaf extract of ALS was obtained fromAmaranthus palmeri. Relative inhibitor potency (I50) values were obtained at saturating substrate conditions for temperatures from 10 to 50 C. Regression analysis of field activity against I50values showed the two data sets to be highly correlated (R2= 0.88). The thermal dependence of enzyme/herbicide interactions may provide another means of understanding environmental factors limiting herbicidal efficacy and predicting herbicide inhibition at the whole-plant level.
Transgenic cotton (Gossypium hirsutum L.) lines expressing the tobacco glutathione S-transferase (GST) Nt107 were evaluated for tolerance to chilling, salinity, and herbicides, antioxidant enzyme activity, antioxidant compound levels, and lipid peroxidation. Although transgenic seedlings exhibited ten-fold and five-fold higher GST activity under normal and salt-stress conditions, respectively, germinating seedlings did not show improved tolerance to salinity, chilling conditions, or herbicides. Glutathione peroxidase (GPX) activity in transgenic seedlings was 30% to 60% higher under normal conditions, but was not different than GPX activity in wild-type seedlings under salt-stress conditions. Glutathione reductase, superoxide dismutase, ascorbate peroxidase, and monodehydroascorbate reductase activities were not increased in transgenic seedlings under salt-stress conditions, while dehydroascorbate reductase activity was decreased in transgenic seedlings under salt-stress conditions. Transgenic seedlings had 50% more oxidized glutathione when exposed to salt stress. Ascorbate levels were not increased in transgenic seedlings under salt-stress conditions. Malondialdehyde content in transgenic seedlings was nearly double that of wild-type seedlings under normal conditions and did not increase under salt-stress conditions. These results show that expression of Nt107 in cotton does not provide adequate protection against oxidative stress and suggests that the endogenous antioxidant system in cotton may be disrupted by the expression of the tobacco GST.
Environmental temperature is a critical factor in the lives of almost all organisms. Plants experience periods of thermal stress related to seasonal patterns of temperature and periodic water deficits. Within the range of non-lethal temperatures, there are a number of thermal effects on metabolism that are a result of the thermal dependence of enzymes. The thermal dependence of enzyme kinetic parameters was used to predict that the efficacy of the herbicide pyrithiobac on Palmer amaranth would be reduced at temperatures outside a 20-34 degrees C thermal application range. This prediction is validated in a controlled environment study described in this paper. Palmer amaranth was grown for 16 days in growth chambers with 34/18 degrees C day/night temperature regime. Pyrithiobac was applied to plants at 18, 27 or 40 degrees C. After 1 h at the application temperatures the plants were returned to the 34/18 degrees C regime for 14 days and post-application biomass accumulation (efficacy) was determined. Dry weight accumulation, as a percentage of untreated controls, was 25, 2.5 and 70% for 18, 27 and 40 degrees C application temperatures. Pyrithiobac efficacy was highest for the application within the thermal application range and significantly reduced at temperatures above and below. The validation of the earlier prediction suggests that temperature-related kinetic limitations on herbicide efficacy may also occur in plants with bioengineered herbicide resistance based on herbicide metabolism. The theoretical aspects of such thermal limitations on herbicide resistance mechanisms are discussed.
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