Identification and characterization of differentially expressed carnation petal cDNAs involved in triggering of hypoxic response
2003
Harutunyan, S.
Although controlled atmosphere (CA) storage has been used commercially for more than 50 years, the mode of action of low oxygen in delaying plant senescence is still unclear. Proof of low oxygen interference with the production and/or action of ethylene has been given in the literature. Furthermore, experimental work indicated that O2 is required if C2H4 is to exert its biological effects. The present study aimed at investigating hypoxia-induced changes of gene expression in carnation petals in early stages of hypoxic treatment.Flowers were treated with 1.5 percent oxygen concetration for 15, 30 and 60 minutes. Total RNA was extracted for fingerprinting using the cDNA-Amplified Fragment Length Polymorphism (cDNA-AFLP) technique. Gene expression profile changes due to hypoxic response were detectable after 30 and 60 minutes. Twenty transcript derived fragments (TDFs) were selected for further in silico characterisation. Ten out of twenty transcript derived fragments (TDFs) that appeared to be induced under hypoxia were sequenced, and all but two yielded similarity to known proteins. Three of them were considered to be involved in hypoxic response; TDF HY 1-2 matched ACC oxidase, an ethylene-forming enzyme in arabidopsis, HY 1-6 matched MYB, a DNA-binding-like protein and HY 2-5 matched a putative heat-shock protein. Northern blot analysis failed to detect expression of any of the TDFs in carnation petals during the time-course of the hypoxic treatment (15m, 30m, 1h, 2h, 3h, 6h, 12h and 24h), probably due to low levels of mRNA abundance of the identified TDFs
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