The crop area planted to conventional soybeans has decreased annually while that planted to glyphosate-resistant (RR) soybean has drastically increased mainly due to the wide adoption of glyphosate in current weed management systems. With the extensive use of glyphosate, many farmers have noted visual plant injury in RR soybean varieties after glyphosate application. A new generation designated as "second generation-RR2" has been recently developed and these RR2 cultivars already are commercially available for farmers and promoted as higher yielding relative to the previous RR cultivars. However, little information is currently available about the performance of RR2 soybean beyond commercial and farmer testimonial data. Thus, an evaluation of different glyphosate rates applied in different growth stages of the first and second generation of RR soybeans, revealed a significant decrease in photosynthesis. In general, increased glyphosate rate and late applications (V6) pronounced decrease photosynthetic parameters and consequently decreased in leaf area and shoot biomass production. In contrast, low rate and early applications were less damage for the RR soybean plants, suggesting that with early applications (V2), plants probably have more time to recover from glyphosate or its metabolites effects regarding late applications.