Background: Moringa oleifera L. (moringa) is a promising plant for nutritionally and medicinally uses. The influence of laser radiation on moringa seeds (dry and wetted) was investigated, using helium-neon (He-Ne) laser source at ʎ = 632.8 nm wavelength and 5 cm beam diameter for 25, 45, and 90 min. Agro-morphological criteria, SDS-PAGE protein banding patterns, and ISSR markers were investigated to elucidate the influence of He-Ne laser on moringa.Results: Most agro-morphological criteria increased especially with wetted seed group pre-laser treatments (25 min). SDS-PAGE and ISSR profiles showed changes which include the absence of some bands and the appearance of few novel bands. ISSR markers showed a change in band number in all investigated samples with a total number of 172 bands. The polymorphic bands were 91 bands with 19 unique bands. The average mean percentage of polymorphism was recorded 52.91% at samples treated with a laser at different times. Wetted seed treatment for 25 min recorded 40 bands as new bands. Genetic template stability (GTS) values were recorded in all treatments. The lowest GTS % was recorded 76.74 at 25 min (wetted seeds) while highest GTS % was recorded 83.72 at 25 min (dry seeds). According to the data of GTS, produced by laser treatment for 25 and 45 min for wetted and dry seeds, respectively, it is more effective on genome stability than other treatments. Conclusion: He-Ne laser treatments of wetted seeds achieve biostimulation in many aspects of agromorphological criteria. ISSR analysis is a highly sensitive method for the detection of DNA alteration induced by laser treatments. The exposure of dry and wetted seeds to He-Ne laser at different times affected the genomic template stability percentage (GTS %). Also, the laser treatments have a great effect on gene expression by switching on or off some genes that may lead to the presence or absence of certain proteins, respectively.