Developmental variation in floral volatiles composition of a fragrant orchid Zygopetalum maculatum (Kunth) Garay.

Emitted scent volatile profile of an orchid species Zygopetalum maculatum was studied using dynamic headspace sampling technique with four different adsorbent matrices, namely Porapak Type Q polymer (mesh size: 80/100), Tenax (mesh size: 60/80), activated charcoal and graphite. In addition, developmental variations in scent emissions and endogenous volatile levels were also investigated. Gas chromatography-mass spectrometry analysis revealed the presence of 21 volatile compounds in the headspace, which was predominantly enriched with benzenoid compounds. Among these benzenoids, o-diethylbenzene and p-diethylbenzene were the major compounds followed by benzyl acetate and methyl salicylate. Among the phenylpropanoid compounds, 2-phenylethyl acetate was the major volatile. However, as compared to benzenoids, the quantity was much lesser, indicating the inclination of phenylalanine flux towards benzenoid pathway. The outcome of this study has the implications in enhancing fragrance and vase life of orchids of the Sikkim Himalaya region and thus may further help to meet the growing market demand.

Characterization and Synergistic Effect of Antifungal Volatile Organic Compounds Emitted by the Geotrichum candidum PF005, an Endophytic Fungus from the Eggplant.

Plant-associated endophytes are recognized as sources of novel bioactive molecules having diverse applications. In this study, an endophytic yeast-like fungal strain was isolated from the fruit of eggplant (Solanum melongena) and identified as Geotrichum candidum through phenotypic and genotypic characterizations. This endophytic G. candidum isolate PF005 was found to emit fruity scented volatiles. The compositional profiling of volatile organic compounds (VOCs) revealed the presence of 3-methyl-1-butanol, ethyl 3-methylbutanoate, 2-phenylethanol, isopentyl acetate, naphthalene, and isobutyl acetate in significant proportion when analyzed on a time-course basis. The VOCs from G. candidum exhibited significant mycelial growth inhibition (54%) of phytopathogen Rhizoctonia solani, besides having mild antifungal activity against a few other fungi. The source of carbon as a nutrient was found to be an important factor for the enhanced biosynthesis of antifungal VOCs. The antifungal activity against phytopathogen R. solani was improved up to 91% by feeding the G. candidum with selective precursors of alcohol and ester volatiles. Furthermore, the antifungal activity of VOCs was enhanced synergistically up to 92% upon the exogenous addition of naphthalene (1.0 mg/plate). This is the first report of G. candidum as an endophyte emitting antifungal VOCs, wherein 2-penylethanol, isopentyl acetate, and naphthalene were identified as important contributors to its antifungal activity. Possible utilization of G. candidum PF005 as a mycofumigant has been discussed based upon its antifungal activity and the qualified presumption of safety status.

Enzymatic production and emission of floral scent volatiles in Jasminum sambac.

Floral scent composed of low molecular weight volatile organic compounds. The sweet fragrance of any evening blooming flower is dominated by benzenoid and terpenoid volatile compounds. Floral scent of Jasminum sambac (Oleaceae) includes three major benzenoid esters - benzylacetate, methylbenzoate, and methylsalicylate and three major terpene compounds viz. (E)-ß-ocimene, linalool and α-farnesene. We analyzed concentrations and emission rates of benzenoids and terpenoids during the developmental stages of J. sambac flower. In addition to spatial emission from different floral parts, we studied the time-course mRNA accumulations of phenylalanine ammonia-lyase (PAL) and the two representative genes of terpenoid pathway, namely 1-deoxy-d-xylulose 5-phosphate reductoisomerase (DXR) and terpene synthase (TPS). Further, in vitro activities of several enzymes of phenylpropanoid/benzenoid pathway viz., PAL and acetyl-coenzyme A: benzylalcohol acetyltransferase (BEAT), S-adenosyl-l-methionine: benzoic acid carboxyl methyl transferase (BAMT) and S-adenosyl-l-methionine: salicylic acid carboxyl methyltransferase (SAMT) were studied. All the above enzyme activities along with the in vitro activities of DXR and TPS were found to follow a certain rhythm as observed in the emission of different benzenoid and terpenoid compounds. Linalool emission peaked after petal opening and coincided with maximal expression of JsTPS gene as evidenced from RT-PCR analyses (semi-quantitative). The maximum transcript accumulation of this gene was observed in flower petals, indicating that the petals of J. sambac flower play an important role as a major contributor of volatile precursors. The transcripts accumulation of JsDXR and JsTPS in different developmental stages and in different floral part showed that emissions of terpenoid volatiles in J. sambac flower are partially regulated at transcription levels.

Biostimulation of Indigenous Microbial Community for Bioremediation of Petroleum Refinery Sludge.

Nutrient deficiency severely impairs the catabolic activity of indigenous microorganisms in hydrocarbon rich environments (HREs) and limits the rate of intrinsic bioremediation. The present study aimed to characterize the microbial community in refinery waste and evaluate the scope for biostimulation based in situ bioremediation. Samples recovered from the wastewater lagoon of Guwahati refinery revealed a hydrocarbon enriched [high total petroleum hydrocarbon (TPH)], oxygen-, moisture-limited, reducing environment. Intrinsic biodegradation ability of the indigenous microorganisms was enhanced significantly (>80% reduction in TPH by 90 days) with nitrate amendment. Preferred utilization of both higher- (>C30) and middle- chain (C20-30) length hydrocarbons were evident from GC-MS analysis. Denaturing gradient gel electrophoresis and community level physiological profiling analyses indicated distinct shift in community's composition and metabolic abilities following nitrogen (N) amendment. High throughput deep sequencing of 16S rRNA gene showed that the native community was mainly composed of hydrocarbon degrading, syntrophic, methanogenic, nitrate/iron/sulfur reducing facultative anaerobic bacteria and archaebacteria, affiliated to γ- and δ-Proteobacteria and Euryarchaeota respectively. Genes for aerobic and anaerobic alkane metabolism (alkB and bssA), methanogenesis (mcrA), denitrification (nirS and narG) and N2 fixation (nifH) were detected. Concomitant to hydrocarbon degradation, lowering of dissolve O2 and increase in oxidation-reduction potential (ORP) marked with an enrichment of N2 fixing, nitrate reducing aerobic/facultative anaerobic members [e.g., Azovibrio, Pseudoxanthomonas and Comamonadaceae members] was evident in N amended microcosm. This study highlighted that indigenous community of refinery sludge was intrinsically diverse, yet appreciable rate of in situ bioremediation could be achieved by supplying adequate N sources.

Inter-specific variation in headspace scent volatiles composition of four commercially cultivated jasmine flowers.

Jasmines are commercially grown for their fragrant flowers and essential oil production. The flowers of jasmine emit sweet-smelling fragrance from evening till midnight. This study was designed to study the composition and inter-specific variation of the emitted scent volatiles from flowers of four commercially cultivated Jasminum species namely, Jasminum sambac, Jasminum auriculatum, Jasminum grandiflorum and Jasminum multiflorum. Gas chromatography-mass spectrometry analysis revealed that the scent volatiles composition of these flowers was predominantly enriched with both terpenoid and benzenoid compounds. Linalool and (3E,6E)-α-farnesene were identified as the major monoterpene and sesquiterpene in all the four species, respectively. The most abundant benzenoid detected in all flowers was benzyl acetate. Comparison of volatile profiles indicated a variation in fragrance contents and types emitted from these four jasmine flowers. The outcome of this study shall help in elucidating the enzymes and genes of fragrance biosynthesis in jasmines and in aiming to create flowers with improved scent quality.