Matoa (Pometia pinnata JR Forst & G Forst) has potential as a biodiesel raw material. Chitosan can increase plant growth by stimulating the biosynthesis of auxin and tryptophan. This research aimed to get the chitosan application time and the matoa type that can increase matoa growth. The method used was a completely randomized design (CRD) with two factors. The first was the application time of chitosan (without chitosan application, morning, afternoon, a combination of morning and evening) and the second was matoa type (red, green, and yellow). Chitosan was given at a concentration of 4%. Data analysis used analysis of variance and continued with Duncan’s Multiple Range Test (DMRT) at the 5% significance level. The results showed that chitosan application in the morning time gave the best results on the photosynthetic rate, the afternoon gave the best results on the height and photosynthetic rate, and the morning and afternoon gave the best results on the stem diameter. The interaction of chitosan application time in the afternoon with green matoa species gave the best results on the plant height and in the afternoon with red matoa and in the morning with green matoa gave the best results on the photosynthetic rate.