Metabolite profiling of Piper longum L. fruit volatiles by two-dimensional gas chromatography and time-of-flight mass spectrometry: Insights into the chemical complexity.

Piper longum L. (long pepper) is an economically and industrially important medicinal plant. However, the characterization of its volatiles has only been analyzed by gas chromatography-mass spectrometry (GC-MS). In the present study, precise characterization of P. longum fruit volatiles has been performed for the first time through advanced two-dimensional gas chromatography-time-of-flight spectrometry (GC×GC-TOFMS). A total of 146 constituents accounting for 93.79% were identified, of which 30 were reported for the first time. All these constituents were classified into alcohols (4.5%), alkanes (8.9%), alkenes (6.71%), esters (6.15%), ketones (0.58%), monoterpene hydrocarbons (1.64%), oxygenated monoterpenes (2.24%), sesquiterpene hydrocarbons (49.61%), oxygenated sesquiterpenes (13.03%), phenylpropanoid (0.23%), and diterpenes (0.2%). Among all the classes, sesquiterpene hydrocarbons were abundant, with germacrene-D (2.87% ± 0.01%) as the major one, followed by 8-heptadecene (2.69% ± 0.03%), ß-caryophyllene (2.43% ± 0.03%), n-heptadecane (2.4% ± 0.04%), n-pentadecane (2.11% ± 0.05%), and so forth. Further, 20 constituents were observed to be coeluted and separated precisely in the two-dimensional column. The investigation provides an extensive metabolite profiling of P. longum fruit volatiles, which could be helpful to improve its therapeutic potential.

Derivatives of Cinnamic Acid Esters and Terpenic Diversity in Volatiles of Thirty-Six Sand Ginger (Kaempferia galanga L.) Accessions of Eastern India Revealing Quality Chemovars.

The essential oil of Kaempferia galanga L. commonly known as sand ginger has increased its demand in national and international market for decades. Cinnamic acid esters like ethyl-p-methoxy cinnamate (EPMC) and ethyl cinnamate (EC) are major constituents in its essential oil. In spite of the high demand for the plant as raw material, identification of quality chemovars having high essential oil (EO) yield and constituents is still at an infant stage. With this in mind, we have evaluated the EO yield of 36 accessions from three provinces of Eastern India, which varied within a range of 0.41 ± 0.01 to 2.63 ± 0.03 v/w. Further, a total of 65 compounds were detected by gas chromatography and mass spectrometry (GC-MS) with area percentages varying from 76.16 to 97.3%. EPMC was found to be the major component in 14 accessions with area percentages varying from 10.7% to 41.1%, whereas other 22 accessions showed EC as the major constituent, varying from 16% to 29.1%. Further, a diversity study among accessions was performed by agglomerative hierarchical clustering (AHC) and principal component analysis (PCA) analysis based on the abundance of identified constituents, which categorized all 36 accessions into three clusters. Thus, the present study helps to identify quality chemovar K.g16 and K.g14 with respect to oil yield and constituents, respectively, which could be used to guide commercial cultivation and further improvement of the taxa.

Potential role of Indian long pepper (Piper longum L.) volatiles against free radicals and multidrug resistant isolates.

In the present study, the extracted volatiles from dried leaf and fruit of Piper longum were analysed by gas chromatography-mass spectrometry (GC-MS) and each detected 53 constituents having 92.41% and 96.31% of the total volatiles respectively. E-nerolidol (19.56%), ß-pinene (17.07%) and α-pinene (6.8%) were main constituents in leaf volatiles whereas the fruit volatiles dominated by germacrene-D (23.38%), 8-heptadecene (8.95%) and ß-caryophyllene (8.20%). Antioxidant potential of the volatiles were assessed by DPPH (1,1-diphenyl-2-picrylhydrazyl) and ABTS (2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid) assays. The fruit volatiles revealed higher radical scavenging activities as compared to leaf. The samples were also evaluated against multidrug resistant (MDR) isolates including one non MDR fungal strain. The fruit volatiles showed a very strong activity against Acinetobacter baumannii than others whereas leaf volatiles possessed strong activity against Klebsiella pneumoniae as compared to other strains. Thus, the dried fruits can be exploited for drug development towards therapeutic purpose.

Thermal desorption modulation based detection of volatile constituents of Alpinia galanga by two dimensional gas chromatography and time of flight mass spectrometry.

Alpinia galanga Wild.(L.) is well known for its aromatic constituents. Though the aromatic composition is already known, but lots of constituents which contributing overall aroma of the oil are still unknown due to the co-eluting factor of single column in GC-MS. Thus the current study aims to characterise maximum volatile constituents present in the essential oil of A. galanga using thermal desorption modulator of two-dimensional gas chromatography and time-of-flight mass spectrometry. The 102 compounds with good match and high probability value were identified out of which 42 were identified for the first time. The total identified compounds include 47 hydrocarbons 25 alcohols, 7 ketones, 7 esters, 3 aldehyde, 4 ethers and 9 other classified aromatic compounds. It was further categorised into Monoterpene Hydrocarbons, Oxygenated Monoterpenes, Sesquiterpene Hydrocarbons and Oxygenated Sesquiterpenes. The major constituent also varies with respect to area percentage. The in-depth characterisation will help in its qualitative analysis.

Rapid plant regeneration in industrially important Curcuma zedoaria revealing genetic and biochemical fidelity of the regenerants.

The present investigation was carried out to establish an efficient and reproducible micropropagation protocol for the production of morphologically, genetically and chemically uniform plants of Curcuma zedoaria. Axillary bud explants of C. zedoaria were inoculated into MS basal medium supplemented with various combinations and concentrations of 6-benzyladenine (2.2-22.2 µM, BA), kinetin (2.3-23.2 µM, Kin), indole-3-acetic acid (2.9-11.4 µM, IAA), α-naphthalene acetic acid (2.7-10.2 µM, NAA) and adenine sulphate (33.9-203.6 µM, Ads). Almost 95% of rhizome buds sprouted on MS medium supplemented with 13.3 µM BA, 5.7 µM IAA and 63.9 µM Ads giving rise to an average of 12.89 ± 0.02 shoots within 6 weeks. However, the maximum number of roots (25.8 ± 0.07 roots per explant) was obtained on half strength MS medium supplemented with 7.4 µM of IBA after 4 weeks of inoculation. Morphological characteristics were similar in both conventionally propagated and micropropagated plants. Additionally, genetic homogeneity of in vitro plants was further confirmed through ISSR and flow cytometry analysis. A total of 27 ISSR primers were screened, out of which 13 ISSR primers generated 58 monomorphic and reproducible bands thereby confirming the genetic uniformity of obtained plants. The mean 2C DNA content of the mother plant (2.96 pg) was similar to that of in vitro derived plants (3.07 pg). Gas chromatography-mass spectrometry (GC-MS) analysis showed similarity in the qualitative profile of chemical constituents of essential oil and high-performance liquid chromatography analysis revealed no significant differences in curcumin content in the tissue culture regenerants and mother plants of C. zedoaria. Therefore, the present micropropagation protocol could be effectively employed to generate true to type plantlets of C. zedoaria.

EST-SSR marker revealed effective over biochemical and morphological scepticism towards identification of specific turmeric (Curcuma longa L.) cultivars.

Turmeric (Curcuma longa L., family Zingiberaceae) is one of the most economically important plants for its use in food, medicine, and cosmetic industries. Cultivar identification is a major constraint in turmeric, owing to high degree of morphological similarity that in turn, affects its commercialization. The present study addresses this constraint, using EST-SSR marker based, molecular identification of 8 elite cultivars and 88 accessions in turmeric. Fifty EST-SSR primers were screened against eight cultivars of turmeric (Suroma, Roma, Lakadong, Megha, Alleppey Supreme, Kedaram, Pratibha, and Suvarna); out of which 11 primers showed polymorphic banding pattern. The polymorphic information content (PIC) of these primers ranged from 0.13 to 0.48. However, only three SSR loci (CSSR 14, CSSR 15, and CSSR 18) gave reproducible unique banding pattern clearly distinguishing the cultivars 'Lakadong' and 'Suvarna' from other cultivars tested. These three unique SSR markers also proved to be effective in identification of 'Lakadong' cultivars when analysed with 88 accessions of turmeric collected from different agro-climatic regions. Furthermore, two identified cultivars (Lakadong and Suvarna) could also be precisely differentiated when analysed and based on phylogenetic tree, with other 94 genotypes of turmeric. The novel SSR markers can be used for identification and authentication of two commercially important turmeric cultivars 'Lakadong' and 'Suvarna'.

Chemical composition and antioxidant activity of essential oil from leaves and rhizomes of Curcuma angustifolia Roxb.

The essential oil extracted from rhizome and leaf of Curcuma angustifolia Roxb. (Zingiberaceae) was characterised by gas chromatography-mass spectrometry (GC-MS). The GC-MS analysis revealed the presence of 32 and 35 identified constituents, comprising 92.6% and 92% of total leaf and rhizome oil, respectively. Curzerenone (33.2%), 14-hydroxy-δ-cadinene (18.6%) and γ-eudesmol acetate (7.3%) were the main components in leaf oil. In rhizome oil, curzerenone (72.6%), camphor (3.3%) and germacrone (3.3%) were found to be the major constituents. Antioxidant capacities of oil were assessed by various methods, 2, 2-diphenyl-1-picrylhydrazyl (DPPH), 2, 2-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and reducing power ability (RPA). Based on the results, the leaf oil showed more antioxidant potential as compared to rhizome oil and reference standards (ascorbic acid and butylated hydroxytoluene (BHT)). Thus, the leaf essential oil of C. angustifolia can be used as an alternative source of natural antioxidant.

Transcriptome profiling of Curcuma longa L. cv. Suvarna.

Turmeric is an economically valued crop, because of its utility in the food, pharmaceutical industries and Ayurvedic medicine, attracts the attention in many areas of research work. In the present study, we executed resequencing through transcriptome assembly of the turmeric cultivar Suvarna (CL_Suv_10). Resequencing of Suvarna variety has generated 5 Gbases raw data with 75 bp paired-end sequence. The raw data has been submitted to SRA database of NCBI with accession number SRR4042181. Reads were assembled using Cufflinks-2.2.1 tool which ended up with 42994 numbers of transcripts. The length of transcripts ranged from 83 to15565, with a N50 value 1216 and median transcript length 773. The transcripts were annotated through number of databases. For the first time transcriptome profiling of cultivar Suvarna has been done, which could help towards identification of single nucleotide polymorphisms (SNPs) between Suvarna and other turmeric cultivars for its authentic identification.

Resequencing of Curcuma longa L. cv. Kedaram through transcriptome profiling reveals various novel transcripts.

Curcuma longa L. (Turmeric), of the family Zingiberaceae, is one of the economically as well as medicinally important plant species. It is a sterile, polyploid and vegetatively propagated spice crop cultivated usually in Southeast Asia. In the current study, we carried out re-sequencing through transcriptome profiling of Curcuma longa cv. Kedaram (Cl_Ked_6). We acquired a total of 1 GB raw data by resequencing through paired-end sequencing using Nextseq 500 platform. The raw data obtained in this study can be accessible in NCBI database with accession number of SRR3928562 with bioproject accession number PRJNA324755. Cufflinks-2.2.1 tool was used for transcriptome assembly which resulted in 39,554 numbers of transcripts. The transcript length ranged from 76 to 15,568, having N50 value of 1221 and median transcript length of 860. We annotated the transcripts using multiple databases. This data will be beneficial for studying sequence variations particularly SNPs between cultivars of turmeric towards authentic identification and discovery of novel functional transcripts in Kedaram.