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Item Plant growth promotion and stress mitigation mediated by the root endophytic fungus Piriformospora indica In Cassava (Manihot esculenta Crantz)(Department of Microbiology, College of Agriculture, Vellayani, 2025-12-12) Akhila P Subhash; Anith, K NThe study entitled “Plant growth promotion and stress mitigation mediated by the root endophytic fungus Piriformospora indica in cassava (Manihot esculenta Crantz)” was conducted in the Department of Microbiology, College of Agriculture, Vellayani, Thiruvananthapuram and in the ICAR–Central Tuber Crops Research Institute (CTCRI), Sreekariyam, Thiruvananthapuram during 2021-2024, with an objective to investigate the effects of the root endophytic fungus P. indica in cassava on productivity, nutrient acquisition and disease suppression against major pathogens. Other microbial agents used in the study were arbuscular mycorrhizal fungus Glomus mosseae, and the biocontrol bacterium Pseudomonas fluorescens PN026. Initially, the pathogens causing stem and root rot disease in cassava were isolated from the naturally infected plants. Three fungal isolates were obtained from the infected cassava samples, and these isolates were identified as Lasiodiplodia theobromae, Fusarium sp. and Cunninghamella elegans with morphological studies and by molecular identification using internal transcribed spacer region (ITS) sequence-based technique. Among them, L. theobromae and Fusarium sp. were identified as the major pathogens and C. elegans as a minor pathogen causing the disease. Pathogenicity of the isolates was verified by proving Koch’s postulates by inoculating the isolated pathogens separately, sequentially and in different combinations in freshly harvested healthy cassava tubers. Treatment with the combination of pathogens L. theobromae and Fusarium sp. was identified as the one showing highest infectivity with high percentage of weight loss and with least number of days taken for the symptom development. This inoculum combination was used for further studies. Direct and indirect antagonism of P. indica against the cassava stem and root rot fungal pathogens was evaluated on potato dextrose agar (PDA) plates. Direct antagonism was evaluated through dual culture plate assay and indirect antagonism through agar well diffusion method using the culture filtrate of P. indica. However, in both the tests P. indica did not inhibit these fungal pathogens in vitro. In vivo studies included cassava root colonization and root initiation, nutrient acquisition and plant growth promotion experiment as well as stem and root rot disease and cassava mosaic disease suppression. An experiment was conducted in protray to evaluate the root colonization abilities of P. indica in Sree Suvarna, Sree Jaya and Aswathy varieties of cassava in both sterile and unsterile planting medium. Fungal structures were detected by trypan blue staining, WGA-AF 488 (Wheat germ agglutinin- Alexa flour 488) staining and confocal imaging. Pear shaped chlamydospores were observed within the root cortical tissues at 14 days after inoculation. Further confirmation was done by molecular detection with the help PiTEF gene primer using a PCR reaction. This is the first report of successful colonization of P. indica in cassava. The root colonization ability of the AM fungus G. mosseae in all the three varieties of cassava in sterile planting medium was also evaluated using trypan blue staining method. The percentage of root colonization was 100 percent in all these varieties, where AM fungal vesicles as well as arbuscles were observed in the roots at 14 days after inoculation. Root initiation tests in minisetts were carried out in Sree Jaya and Aswathy cassava varieties by applying P. indica and Pseudomonas fluorescens in both sterile and unsterile planting medium. When P. indica and P. fluorescens colonized plants were compared to control plants, all root growth parameters were observed to be higher. However, most of parameters were found significant in unsterile planting medium when compared to the sterile planting medium which may be due to the insufficient root colonization of both the beneficial microbes in cassava minisetts in the sterile medium. A field experiment was conducted by applying P. indica and AM fungus G. mosseae, along with different doses of NPK fertilizers in cassava variety Sree Suvarna to evaluate the abilities of both beneficial fungi to promote growth and acquire nutrients. Different biometric, physiological, yield as well as quality parameters were analyzed. The application with a combination of 75 percent of the recommended dose of NPK along with P. indica was found to be the best treatment that improved growth and yield attributes, whereas the combination of 75 percent NPK along with G. mosseae recorded significant improvement in total dry matter contents as well as tuber quality parameters. The results showed that the chemical fertilizer dose could be reduced to a level of 75 percent of the recommended dose, without compromising the growth and yield, if the fertilizers are applied together with the bio-inoculants, either P. indica or G. mosseae. The same treatments also resulted in higher total nitrogen uptake while the combination of 100 percent NPK and G. mosseae resulted in better phosphorous and potassium uptake by the cassava plants. A grow bag experiment was conducted to examine the stem and root rot disease suppressing ability of P. indica in the local disease-prone cassava variety Aswathy. The treatment combination of pathogens, L. theobromae and Fusarium sp. was selected for the inoculation purpose. An increase in the growth and yield and decrease in disease incidence was noted in P. indica and G. mosseae inoculated plants, even when the plants were challenged with the stem and root rot pathogens. The pathogen inoculated control plants recorded the least number of days taken for the initial symptom development (6 months after planting), highest percentage of tubers affected per plant (53.33%) and extent of damage in each tuber (50%) followed by the chemical control plants. Simultaneously another grow bag experiment was also conducted, in the mosaic susceptible variety Sree Jaya to assess the cassava mosaic virus disease suppression by P. indica. Natural incidence of cassava mosaic virus in the cassava plants was monitored by visual scoring from 4 MAP till harvest (7 MAP). The plants from absolute control recorded highest visible symptoms of mosaic disease which was on par with G. mosseae treated plants, and least symptoms were recorded in the P. indica treated plants. Viral load in the cassava leaves estimated by quantitative polymerase chain reaction (qPCR) showed highest viral copy number in the G. mosseae treated plants and least in the P. indica treated plants. However, the G. mosseae treated plants showed maximum yield parameters compared to all other treatments. From the results of the present study, it can be inferred that the combined application of P. indica (vermiculite based inoculum one percent w/v @ 20 g/pit) or AM fungus G. mosseae (@ 5 g/pit) along with 75 percent recommended dosage of NPK fertilizers at the time of planting helps to reduce the amount of chemical fertilizers as well as enhance the growth and yield parameters of cassava plants. The study also suggests that P. indica as well as AM fungus G. mosseae can be utilized as biocontrol agents to combat stem and root rot disease of cassava. Effective colonization of P. indica also reduces the incidence of cassava mosaic virus. The combined application of these two beneficial endophytic fungi might have the potential to improve the growth and yield parameters of cassava with positive effects on disease control.