Project description:The project is aimed at analysing the comparitive proteomics of red rot pathogen, C. falcatum, during red rot infection in sugarcane. The differentially abundant proteins shall be used to identify the corresponding genes.
Project description:Comparative transcriptome analysis of virulent and avirulent strains of Colletotrichum falcatum to unravel the pathogenic variations.
Project description:A total of 226 sugarcane rhizosphere-associated bacterial strains from the six different cultivars were screened against three pathogenic strains of C. falcatum (cfNAV, cfCHA, and cf8436) for the suppression of red rot disease. On the basis of mycelial growth inhibition in dual culture assay, 26 bacteria were selected for further characterization of morphology, biochemical activity, plant-growth-promoting (PGP) activity, antifungal potential and molecular identity by 16S rRNA gene sequence. On the basis of the 16S rRNA gene sequencing, it was found that the isolates belonged to proteobacteria (13), Firmicutes (10), and Bacteroides (3). The antagonistic bacteria tested for PGP traits revealed that 10 strains were able to solubilize tricalcium phosphate, 11 strains were able to produce siderophore, and 14 strains were able to grow in the N-free medium. The quantitative estimation of indole-3-acetic acid production was ranged from 21.58 to 66.31??g/mL. On the basis of PGP and biocontrol traits, five strains Ochrobactrum intermedium (TRD14), Acinetobacter sp. (PK9), Bacillus sp. (RSC29 and KR91) and Escherichia sp. (VRE34) were further chosen for pot trial under greenhouse conditions on highly susceptible variety CoC671. The results showed that the pathogen-inoculated sugarcane plants were able to germinate but died within one month. However, the CoC671 inoculated with selected biocontrol strains found protected from disease and an increase in plant growth parameters on par with carbendazim fungicides. This study proves that the isolates identified in this study could be used as an alternative to chemical fungicides to control red rot pathogen of sugarcane plants.
Project description:We evaluated transgenic lines of sugarcane modified with the barley chitinase class-II gene to create resistance against the red rot causative agent Colletotrichum falcatum Went. Local sugarcane cultivar SP93 was transformed with a 690-bp coding sequence of the chitinase-II gene under the influence of a polyubiquitin promoter. Transgenic sugarcane lines (T 0) overexpressing the chitinase gene were obtained through a particle bombardment method with 13.3% transformation efficiency. Four transgenic sugarcane lines, SCT-03, SCT-05, SCT-15, and SCT-20, were tested for resistance against red rot by in vitro antifungal assays. Crude protein extracts from transgenic sugarcane plants SCT-03, SCT-05, SCT-15, and SCT-20 inhibited the mycelial growth of C. falcatum by 49%, 40%, 56%, and 52%, respectively, in a quantitative in vitro assay. Our findings revealed that two transgenic lines, SCT-15 and SCT-20, exhibited the highest endochitinase activity of 0.72 and 0.58 U/mL, respectively. Furthermore, transgenic lines SCT-15 and SCT-20 exhibited strong resistance against inoculated C. falcatum in an in vitro bioassay, as they remained healthy and green in comparison with the control sugarcane plants, which turned yellow and eventually died 3 weeks after infection. The mRNA expression of the transgene in the C. falcatum-inoculated transgenic sugarcane lines increased gradually compared to the control plant. The mRNA expression was the highest at 72 h in both transgenic lines and remained almost stable in the subsequent hours.