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

Abstract

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 butenolidetreated seedlings were not significantly different from each other according to the non-parametrical KruskalWallis 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.

Genotoxicity evaluation of two kinds of smoke‐water and 3,7‐dimethyl‐2H‐furo[2,3‐c]pyran‐2‐one

“…Preliminary testing against the TA100 strain with higher doses of the concentrated crude smoke solution showed that these doses were just below toxicity levels. The abovementioned negative results for the genotoxic potential of smokewater or the active butenolide compound were confirmed by Jain et al (2008), who reported on changes in DNA, RNA and protein during seed germination in tomato, following the use of 3-methyl-2H-furo[2,3-c]pyran-2-one.…”

Regulation of Seed Germination and Seedling Growth by Chemical Signals from Burning Vegetation

“…Differential display of complementary DNA (cDNA) from Solanum esculentum (tomato; Solanaceae) seed treated with KAR 1 for 72 h led to the tentative identification of four transcripts, including an expansin gene; however, no confirmation of differential expression was performed (54). Another investigation using fluorescent differential display of lettuce seed treated with SW in the dark found 11 cDNAs with altered expression at one or more time points during seed imbibition and early germination.…”

Effects of aqueous smoke solutions and heat on seed germination of herbaceous species of the Sudanian savanna-woodland in Burkina Faso

“…Smoke affects plant processes through interaction, in some way, with endogenous plant growth regulators (van Staden et al, 2000), and the active compound (karrikinolide) could be involved in early induction of the cell cycle activities, thus accelerating radicle emergence in germinating seeds (Jain and Van Staden, 2006). Karrikinolide has now been suggested to increase the expression levels of expansins (Jain et al, 2008a), proteins mainly involved in cell wall loosening activity, required during the process of seed germination to reduce the physical restriction that the endosperm causes to the embryo expansion. With respect to post-germination smoke effects such as increasing seedling vigour, the karrikinolide compound has been shown to have both cytokinin-and auxin-like activity when applied at low concentrations (Jain et al, 2008b).…”