The recent investigation was carried out to determine the effect of different gamma-ray doses and 900 W (2450 MHz) microwave radiation with various exposure times, separately or in combinations, on the yield, yield components, and chemical properties of guar (Cyamopsis tetragonoloba), as well as, to detect variation induction. The cDNA-SCoT technique was used to obtain molecular markers related to some traits. SSR technique was used to sequence the target fragment related to plant height. Gamma-ray doses of 150 and 250 Gy alone, and in combination with 900 W microwaves irradiation applied with different duration or time span (1, 2, 3, and 4 min) influenced the plant height significantly, as well as, number of tillers plant-1 and fresh and dry forage yield, and fresh and dry leaf stem-1 ratio. In the second sample, seed yield at harvest time, e.g., pods plant-1, weight of pods plant-1, whole plant dry weight, number of seeds pod-1, length of pod, 100-seed weight, and seeds yield were affected by irradiation with different and varied responses. In the M1 generation, the 18 SCoT primers produced 327 bands ranging between 151–2895 bp in size, out of which 282 were polymorphic (86.24%). In the M2, the 18 SCoT primers produced 328 bands ranging between 212–2661 bp in size, out of which 299 were polymorphic (91.16%). The M1 and M2 generations exhibited 89 positive and 39 negative bands, which could be used as marker assisted-selection in response to treated guar plants with different gamma ray doses, separately or in combinations with microwave treatments.