Transcriptional reprogramming and enhanced photosynthesis drive inducible salt tolerance in sugarcane mutant line M4209.

Sugarcane (Saccharum officinarum) is a globally cultivated cash crop whose yield is negatively affected by soil salinity. In this study, we investigated the molecular basis of inducible salt tolerance in M4209, a sugarcane mutant line generated through radiation-induced mutagenesis. Under salt-contaminated field conditions, M4209 exhibited 32% higher cane yield as compared with its salt-sensitive parent, Co86032. In pot experiments, post-sprouting phenotyping indicated that M4209 had significantly greater leaf biomass compared with Co86032 under treatment with 50 mM and 200 mM NaCl. This was concomitant with M4209 having 1.9-fold and 1.6-fold higher K+/Na+ ratios, and 4-fold and 40-fold higher glutathione reductase activities in 50 mM and 200 mM NaCl, respectively, which suggested that it had better ionic and redox homeostasis than Co86032. Transcriptome profiling using RNA-seq indicated an extensive reprograming of stress-responsive modules associated with photosynthesis, transmembrane transport, and metabolic processes in M4209 under 50 mM NaCl stress. Using ranking analysis, we identified Phenylalanine Ammonia Lyase (PAL), Acyl-Transferase Like (ATL), and Salt-Activated Transcriptional Activator (SATA) as the genes most associated with salt tolerance in M4209. M4209 also exhibited photosynthetic rates that were 3-4-fold higher than those of Co86032 under NaCl stress conditions. Our results highlight the significance of transcriptional reprogramming coupled with improved photosynthetic efficiency in determining salt tolerance in sugarcane.

Radiation induced mutagenesis, physio-biochemical profiling and field evaluation of mutants in sugarcane cv. CoM 0265.

PURPOSE: Sugarcane is an important cash crop and is affected by soil salinity. CoM 0265, a moderately salt-tolerant variety grown in the Maharashtra region (India), has low sugar content. The present study was aimed to employ gamma ray induced in vitro mutagenesis with repeated and step-wise selection in sugarcane for the isolation and physio-biochemical profiling of the selected salt-tolerant mutants for improved agronomic performance and sugar content. MATERIALS AND METHODS: Embryogenic callus culture of CoM 0265 variety was subjected to different doses of gamma radiation (10, 20, 30, 40, 50, and 60 Gy) followed by selection on NaCl containing media (50, 100, 150, 200, and 250 mM NaCl). The regenerated plantlets were hardened and selected based on ground nursery field trial on normal soil and saline field trial, in augmented block design for the selected mutant clones. Different physio-biochemical changes and activity of antioxidant enzymes were analyzed in the salt selected in vitro cultures and field-grown mutant clones. RESULTS: Dose optimization showed 40 Gy as the LD50 for gamma radiation and 150 mM NaCl as the dose for in vitro selection experiments. The selected mutant clones showed higher tissue water content (TWC), chlorophyll, and lower sodium content indicative of tolerance to salt stress. Catalase and peroxidase enzyme activities in the top visible dewlap (TVD) of the putative mutant clones were significantly higher than the control. The average yield and sucrose percent of the selected mutant clones were significantly higher than control checks in the saline field trial. Mutant clones M8457 and M8721 exhibited improved yield and commercial cane sugar over the parent control check varieties under saline field conditions. Catalase activity was strongly associated with TWC (r = 0.34) and chlorophyll content (r = 0.41) while it was negatively correlated with sodium ion content (r = -0.38). Peroxidase activity in TVD also showed a significant positive correlation with chlorophyll content (r = 0.42) and a negative correlation with sodium ion content (r=-0.39). The improvement in yield and CCS (t/ha) was strongly associated with the lower sodium ion content of the mutant clones (r=-0.54 and -0.53, respectively). CONCLUSIONS: Gamma ray induced mutants were isolated for improved sucrose and high yield in sugarcane var. CoM 0265. The results suggest that gamma radiation induced mutations result in physiological and metabolomic alterations for better growth and adaptation under in vitro and field stress conditions in sugarcane. The improved mutants can be further useful for commercial cultivation in saline areas.