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Published on October 27, 2006; 10.1104/pp.106.087783

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Received August 1, 2006
Accepted October 22, 2006

Evaluation of Source Leaf Responses to Water-Deficit Stresses in Cotton using a Novel Stress Bioassay

John J. Burke *

USDA Plant Stress and Germplasm Development Unit, 3810 4th Street, Lubbock, Texas 79415

* Corresponding author; email: jburke{at}lbk.ars.usda.gov.

Water-deficit stresses preferentially reduce shoot growth, thereby disrupting the flow of carbohydrates from source leaves to the developing sinks. Here, we use a novel stress bioassay to dissect responses of field and greenhouse-grown cotton (Gossypium hirsutum L.) source leaves to water-deficit stresses. Fifth main stem leaf samples were harvested at sunrise and subjected to a prolonged elevated respiratory demand in the dark. Sucrose levels are lower in non-stressed cotton at sunrise compared to water-deficit stressed cotton, potentially predisposing the non-stressed tissue to succumb more rapidly. Tissue death was determined initially using the cell viability stain 2,3,5-triphenyltetrazolium chloride, but was determined in subsequent experiments by monitoring the decline in chlorophyll fluorescence yield. Fluorescence yield measurements were obtained within minutes of harvesting and individual samples were monitored over the time course of the treatment. Analyses of the time course and magnitude of chlorophyll fluorescence yield decline in samples from irrigated and dryland plots permitted the detection of stress responses within 24 h of the cessation of irrigation. The rate of fluorescence yield decline during the elevated respiratory demand treatment slowed as the water-deficit stress increased. Upon irrigation, the source leaves of the water stressed plants recovered to pre-stress values within 4 days. Well-watered cotton over-expressing hsp101 had identical rates of fluorescence yield decline as non-transgenic cotton. These results suggest that the delayed decline in fluorescence yield of water stressed tissue exposed to prolonged elevated respiratory demand can be used as a sensitive indicator of water-deficit stress responses.






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