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Isolation and Molecular Characterization of the Ethylene Receptor ETR1 from 'Carabao' Mango (Mangifera indica L.) Fruit
Thesis Abstract:
Signal transduction of ripening in climacteric mango is controlled by an ethylenespecific receptor protein in the plasma membrane of plant cells. To regulate ripening in mango by manipulating ethylene sensitivity, the gene encoding receptor protein was isolated and characterized.
Partial cDNAs corresponding to ETR1 homologous have been cloned from ripening mesocarp tissues derived from mango cv. Carabao using reverse transcription followed by polymerase chain reaction (RT-PCR). The primers ATGGCC(A/T) GCTGATG (forward) and ATAAAACGCTCCTC (reverse) were designed based on multiple sequence alignment analysis of ETR1 genes available at the GenBank database. Total RNA isolated from ripening mesocarp tissues of mango was used as template for RT-PCR and generated at least two types of ETR1 homologous (1.1 and 1.7 kb PCR fragments). These two ETR1 homologous were cloned in TOPO-TA vectors for DNA sequencing. Sequence analysis of the partial cDNAs (Fragments A1, A2, B to F) revealed that the ethylene receptor in mango belongs to the ETR1- like subfamily (Type I) with three hydrophobic domains at the sensor region of the receptor.
The partial ripening-related cDNA sequences of ETR1 homologous from mango cv. Carabao could be used for manipulating ethylene sensitivity or perception by antisense or co-suppressions technology and interference RNA (iRNA). This study represented the first DNA sequence for an ethylene-specific receptor protein involved in the ripening of the climacteric mango cv. Carabao.