Oligosaccharides, as a wide type of polysaccharide, have a broad antimicrobial spectrum and promote development as plant growth stimulants. To investigate the regulation effects of different oligosaccharides on the dynamic changes of chlorophyll content, leaf fluorescence, root activity and morphology, and chloroplast ultrastructure, as well as the yields and yield components of strawberry and cucumber, typical greenhouse experiments were conducted over two years (2021–2022). The experimental plants were foliar sprayed with tap water (CK), chitosan oligosaccharide (CSOS), and mixed oligosaccharides (MixOS) five times before flowering. The conventional management (CM) was conducted as a conventional control. The findings of the present study suggest that the application of MixOS has the greatest regulation effects on delayed leaf senescence, well-developed roots, and higher fruit productions of strawberry and cucumber. Exogenous MixOS resulted in significant increases in SPAD values, maximum photochemical efficiency (Fv/Fm), and photochemical quenching coefficiency (qP); they were increased by 1.94–28.96%, 5.41–33.89%, and 9.93–62.07%, compared to the CSOS, CM, and CK treatments, respectively. The orderly and steady structure of thylakoids in the chloroplast, and the randomly distributed starch grains, could be clearly observed in the MixOS treatment, while the non-photochemical quenching (NPQ) was correspondingly reduced by 19.04–45.92%. Meanwhile, the remarkable promotion of root activity and root surface morphology indicators (i.e., root length, surface area, average diameter, and volume) could be observed when exposed to the MixOS treatments, and the total yields of strawberry and cucumber were all increased by 12.40–25.57%. These findings suggest that the mixed oligosaccharides mainly promote the coordinated growth of root and shoot, which leads to the improved yields of strawberry and cucumber.
