Genome sequencing of Colletotrichum gloeosporioides ES026 reveals plausible pathway of HupA.

BACKGROUND: Colletotrichum gloeosporioides ES026, isolated as an endophytic fungal strain, was found to produce the important medicinal compound HuperzineA (HupA). In a genetic context, ES026 showed potential in elucidating the biosynthetic pathway of HupA. METHODS AND RESULTS: The ES026 strain was sequenced using de-novo Illumina sequencing methods in this study. Assembling the cleaned data resulted in 58,594,804bp, consisting of 404 scaffolds. The G + C mol % content of this genome was 52.53%. The genome progressive-alignment with other 4 Colletotrichum strains revealed that ES026 showed closer relation with 030206, SMCG1#C and Nara gc5. More than 60 putative biosynthetic clusters were predicted with the fungal version antiSMASH4.0 program. More than 33 types I polyketide-related biosynthetic gene clusters were distributed, containing PKS and PKS-NRPS (polyketide-nonribosomal peptides) hybrid gene clusters. Another 8 NRPS biosynthetic gene clusters were distributed among the genome of ES026. The prenyltransferases, probably involved in aromatic prenyl-compounds and terpenoid biosynthesis, were analyzed using bioinformatics tools like MEGA. CONCLUSION: We predicted a new possible biosynthetic pathway for the HupA from the pipecolic acid, based on the published HupA biosynthesis proposed pathway, the biosynthesis and pipecolic acid-derived compounds. We hypothesize that a hybrid PKS-NRPS mega-enzyme was probably involved in the biosynthesis of HupA with the pipecolic acid, the building block of rapamycin, as a HupA precursor. The rapamycin is produced from a polyketide biosynthesis pathway, and the domain incorporating the pipecolic acid is studied.

Evaluation of Bacterial Perpetuation Assays and Plant Biomolecules Antimicrobial Activity against Cotton Blight Bacterium Xanthomonas citri subsp. malvacearum; An Alternative Source for Food Production and Protection.

Cotton (Gossypium hirsutum) is a global cash crop which has gained importance in earning foreign exchange for each country. Bacterial blight caused by Xanthomonascitri subsp. malvacearum (Xcm) has been a seriousdisease in Pakistan's cotton belt on multiple occasions. Bacterium was isolated and identified through various biochemical and diagnostic tests wherehypersensitivity reaction, Gram staining, KOH (potassium hydroxide), catalase, starch hydrolysis, lecithinase and Tween 80 hydrolysis tests confirmed bacterium as Gram-negative and plant pathogenic. Xcm perpetuation assays wereevaluated on various cotton varieties under glasshouse conditions in completely randomized design by three different methods, wherein the scratch method proved to be the best upon CIM-496 and showed 83.33% disease incidence as compared with the other two methods, where Bt-3701 responded with 53.33% incidence via the spray gun method, and 50% with the water splash method on CIM-616, as compared with the control. Similarly, for disease severity percentage, Bt-3701 was pragmatic with 47.21% through scratch method, whereas, in the spray gun method, 45.51% disease severity was noted upon Bt-802, and 31.27% was calculated on Cyto-179 through the water splash method. Owing to the unique antibacterial properties of aqueous plant extracts, the poison food technique showed Aloe vera, Mentha piperita, Syzygiumcumini and Azadirachta indica with 17.77, 29.33, 18.33 and 20.22 bacterial colonies counted on nutrient agarmedium petri plate, respectively, as compared with the control. Measurement of the inhibition zone by disk diffusion technique showed Mentha piperita, Syzygiumcumini, Citrus limon, Moringa oleifera and Syzygium aromaticum to present the most promising results by calculating the maximum diameter of the inhibition zone, viz., 8.58, 8.55, 8.52, 8.49 and 8.41 (mm), respectively, at the highest tested concentration (75 ppm, parts per million) compared with the control. It is probable that the decoction's interaction with the pathogen population on the host plant will need to be considered in future experiments. However, at this moment, more research into the effective management of cotton bacterial blight by plant extracts in terms of concentration determination and development of biopesticides will provide future avenues to avoid environmental pollution.