The cold tolerance studies were carried out in a bi-parental F2 population of a cross between tolerant and susceptible parents (SKUA-529 and HEERA, respectively). The purpose was to screen the individuals of a population for primary cold-tolerance-related attributes. The information generated has a direct application and use in identifying cold tolerance quantitative trait loci (QTLs) and further can be used for genotyping with an appropriate marker system. The screening was carried out on F2-derived F3 seeds and F3 plants for seedling and agronomic traits, respectively. Two tests measuring cold tolerance were conducted. In experiment I, seeds were germinated for 28 days at 13 °C and 7 days at 28 °C, and in experiment II, the seeds were germinated for 72 h at 28 °C, 96 h at 13 °C, and once more for 72 h at 28 °C. Coleoptile length, germination percentage, and radical reduction percentages were all measured in experiment I. The radicle and coleoptile regeneration in experiment II were measured after the cold period. The improvement in cold tolerance was achieved through radicle regrowth, as evidenced by the difference between the second and first measurements. The individual lines from the F2:3 population that recorded high germination (%) were #21, #13, #14, and #15. The percentage of coleoptile length (PERCOL %) was observed to be between the ranges of 23.33% to 53.00%. The reduction in coleoptile length (REDCOL %) was also obtained, and there was less reduction in #15, #16, and #14 and it had a range between 38.46% and 75%. Radicle regrowth (REDRAG) was high at 13 °C in #7, #11, #30, #35, and #36. Survival of the seedling range was between 33.33% and up to 100%, and the highest survival rate was observed in #16. The main objective of this rotation in temperature was to emulate field conditions where there has been a drop in temperature. The evaluations were done for primary cold stress tolerance traits, and it was found that most of these traits exhibited high variability. The mapping population developed may be utilized to generate a linkage map and locate QTLs for tolerance to cold stress in rice. Further, the identified donors for cold tolerance may be utilized for breeding programs aimed at the transfer of low-temperature stress tolerance into susceptible backgrounds. In general, a genotype with improved seedling germination rates, growth rates, and leaf yellowing scores; high seedling survival; lesser reduction in coleoptile length and in radicle development; and recovery following a cold shock at the seedling stage demonstrated its cold resistance. Genotypes with a low germination percentage, a greater number of days to germination, slow growth rate and higher leaf yellowing score, high reduction in coleoptile and radicle growth, and reduced seedling survival indicated cold susceptibility.
