GC-MS-based metabolomics analysis unravels the therapeutic potential of Neolamarckia cadamba fruit peel

Kadam (Neolamarckia cadamba (Roxb.) is an evergreen tropical tree widely grown in Asia, particularly in India. Neolamarckia cadamba commonly known as kadam, cadamba or burflower tree. The roots, leaves, barks, and fruits of N. cadamba possess medicinal properties and are commonly used in the pharmaceutical industry. Fruit peels are the main waste and may contain various biologically active compounds. However, no prior knowledge about the therapeutic compounds of the peel. The objective of the present study was to unveil therapeutic compounds from the peel by Gas Chromatography–Mass Spectrometry (GC-MS) based metabolomics analysis. Metabolites from the kadam fruit peel were isolated and derivatized using MSTFA, characterized by the GC-MS analysis. Raw spectral data were pre-processed, and peak identification was performed using SHIMADZU Postrun analyse software. The metabolites in N. cadamba fruit peel were identified by comparing the peaks with the mass spectral reference database NIST v20. The results showed that the peel of kadam fruit contains 149 metabolites, which were further categorized into 46 different metabolite classes, with 52 different metabolic pathways and 63 biological functions. The principal roles of the metabolites were identified by functional annotation and enrichment analysis. It revealed that metabolites were responsible for anti-inflammation, anti-oxidant, anti-microbial, and anti-cancer properties. In summary, the peel of kadam fruit also contains various therapeutic compounds like other cadamba parts (i.e., roots, leaves, barks, and fruits). Further, comparing the peel with other parts discloses the peel-specific metabolites. The results obtained in this study could be useful for the pharmaceutical industry

Detection of "Candidatus Liberibacter asiaticus" Using Conventional PCR and Nested-PCR and Identification of Primer Sensitivity through Q-PCR in HLB Infected Mandarin Orange and Acid Lime Plants in Tamil Nadu

One of the most destructive citrus diseases in the world is called citrus Huanglongbing (HLB). For effective HLB associated ‘Candidatus liberibacter spp.’ prevention, sensitive and precise diagnostics are essential. Candidatus Liberibacter asiaticus (CLas), Candidatus Liberibacter africanus (CLaf), and Candidatus Liberibacter americanus (CLam) are among the ‘Candidatus liberibacter spp.’ that infect citrus. The most common of these is the species known as Las. To assess the impact of citrus greening disease in the citrus orchard, leaf samples exhibiting greening symptoms were picked up from 210  mandarin orange trees from nine localities in four districts  and among them 189 plants showed HLB positive and leaf samples are picked up from 216 acid lime trees from nine localities in six districts of Tamil Nadu where citrus is cultivated in a larger area and among them 202 plants showed HLB positive. A “conventional singleplex polymerase chain reaction (PCR) Nested PCR and Q-PCR (Quantitative PCR)” was performed on the extracted DNA samples to detect Ca. Liberibacter asiaticus. Among all the mandarin orange growing localities from “Kanalkadu and Thandikudi in Dindigul district” and among the acid lime fields, the samples from  “Sankarankoil from Tirunelveli district and Kallar from Nilgiris district” showed consistent amplification of 1160 base pairs fragment by using specific primers OI1 and OI2C and also 703 base pairs fragment with gene-specific primers A2 and J5  targeting beta-operon (rpiKAJL-rpoBC) gene which is a  ribosomal protein gene of Ca. Liberibacter asiaticus, these results showed that these districts are exclusively positive to Clas . The nested PCR method significantly increased the sensitivity to identify Las up to 10 times and 100 times, respectively, compared to qPCR and conventional PCR in this study. A set of nested PCR primer pairs were examined to diagnose Las. Nested PCR was used to analyse 14 samples from 2 different citrus cultivars in 2 different districts. Among those again kanalkadu orchard for mandarin orange and Sankankoil orchard for acid lime showed more disease incidence and consistent amplification.  The findings indicate that all the  samples tested positive for HLB; the samples of blotched and chlorotic leaves from the acid lime field at Sankarankoil (thirunelveli district) and mandarin orange field at kanlakaadu (Dindigul district)  had the highest positive detection rate (100%).  The outcomes show that the nested PCR primer pairs are capable of detecting Las in a variety of symptomatic tissues, citrus cultivars, and geographical locations. The set of nested PCR primers used in the current work will be a very helpful addition to the existing methods for Las detection