Changes in cellular macromolecules (DNA, RNA and protein) during seed germination in tomato, following the use of a butenolide, isolated from plant-derived smoke

A butenolide, 3-methyl-2H-furo[2,3-c]pyran-2-one, isolated from plant-derived smoke is responsible for the stimulatory role of smoke or aqueous smoke extracts on seed germination. The present study aimed at determining the changes induced by the butenolide at the level of macromolecules (DNA, RNA and proteins) during seed germination. To ascertain changes at the level of DNA, 25 random decamer primers were used. DNA was isolated from 2-week-old seedlings raised in water or butenolide. Amplification products generated ranged in size from 250 bp to 2,500 bp, while in number they ranged from 1 to 9. Ninety-one bands were scored with 17.6% of polymorphism. Total number of bands recorded for 25 primers in control and butenolide-treated seedlings were not significantly different from each other according to the non-parametrical Kruskal-Wallis test. Differential display was used to isolate differentially expressed genes. A total of 48 differentially expressed cDNAs were observed, out of which 20 were cloned into pGEM-T vectors. Alignment of sequences of these cloned products revealed that several bands isolated from differential display gels contained multiple sequences. Four sequences were tentatively identified, while the remaining sequences did not show any significant homology. Electronic homology revealed identity with expansin, photosystem I P700 apoprotein and polyproteins. Similarity index (SI) values for the banding patterns of SDS-PAGE gels was 65.2% for embryos isolated from seeds imbibed either with water or butenolide for 36 h. SI values indicate that the butenolide did not alter the protein profiles drastically. Results of the present study revealed that butenolide does not affect the integrity of DNA. However, it does play some role during transcription and translation.