Abscisic acid (ABA) accumulation in leaves of drought-stressed coconut palms and its involvement with stomatal regulation of gas exchange during and after stress were investigated. Two Brazilian Green Dwarf coconut ecotypes from hot/humid and hot/dry environments were submitted to three consecutive drying/recovery cycles under greenhouse conditions. ABA accumulated in leaflets before significant changes in pre-dawn leaflet water potential ( PD ) and did not recover completely in the two ecotypes after 8 days of rewatering. Stomatal conductance was influenced by ABA under mild drought and by PD under severe drought. There were no significant differences between the ecotypes for most variables measured. However, the ecotype from a hot/dry environment showed higher water use efficiency after repeated cycles of water stress.
ABSTRACT. Many cell functions are redundantly executed in cells, and the experimental approaches that analyze the group of proteins, whose expression is modified in a specific functional condition, enable the identification of the group of proteins that are expressed under stress conditions. The objective of the present study was the evaluation of the genetic expression induced by cadmium (Cd) in Genipa americana L. (Rubiaceae) plants cultivated in nutritive solution, in order to help further studies concerning its use as a plant phytoremediator of such a metallic element. Plants were exposed to increasing concentrations of Cd (0.5, 1, 2, 4, 8, and 16 mg/L), together with the control, in nutritive solution. After the application of the treatments, root tips were harvested for the construction of a cDNA library. Of the 165 expressed sequence tags (ESTs) generated with the construction of the cDNA library, 81 showed homology to genes deposited in the NCBI database, 67 did not show similarity to any available gene, and 17 ESTs ESTs of genes in G. americana exposed to cadmium demonstrated homology with unknown genes. Of the most abundant cDNAs, 16 ESTs were similar to sequences of metallothionein genes. The analysis of ESTs, obtained from the root of G. americana through the construction of a cDNA library, allowed the identification of genes probably associated with proteins and enzymes related to the defense mechanisms of plants when they undergo biotic and abiotic stresses.
ABSTRACT. In Brazil, most cocoa bean production occurs in Southern Bahia. Witches' broom disease arrived in this area in 1989 and has since caused heavy losses in production. The disease is caused by the basidiomycete fungus Moniliophthora perniciosa, a hemibiotrophic fungus that produces the necrosis and ethylene-inducting protein (MpNEP2) during infection; this protein can activate cysteine proteases and induce programmed cell death. Cysteine proteases can be modulated by cystatin. In this study, we overexpressed TcCYS4, a cocoa cystatin, in tobacco plants and evaluated the effect on MpNEP2 in model plants. Tccys4 cDNA was cloned into the pCAMBIA 1390 vector and inserted into the tobacco plants via Agrobacterium tumefaciens. Transgene expression was analyzed by reverse transcription-quantitative PCR and Western blot analysis. Transcript and protein levels in Tcccys4:tobacco lines were 8.9-and 1.5-fold higher than in wild-type plants (wt). Tcccys4:tobacco lines showed no change in growth compared to wt plants. CO 2 net assimilation (A) increased in Tcccys4:tobacco lines compared to wt plants. Only one line showed statistically significant stomatal conductance (gs) and transpiration rate (E) changes. MpNEP2 was infiltered into the foliar mesophyll of Tcccys4:tobacco lines and wt plants, and necrotic lesions were attenuated in lines highly expressing Tccys4. Our results suggest that cocoa cystatin TcCYS4 affects MpNEP2 activity related to the progression of programmed cell death in tobacco plants. This may occur through the action of cystatin to inhibit cysteine proteases activated by MpNEP2 in plant tissues. Further studies are necessary to examine cystatin in the Theobroma cacao-M. perniciosa pathosystem.
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