Abstract: In vitro assessments of chlorophyll content are expensive, laborious and time consuming. The alternative methods which are more rapid and straightforward could be very useful. The aim of this study was to investigate the effects of water stress on chlorophyll content and SPAD Chlorophyll Meter Reading (SCMR) and the relationship between chlorophyll content and SCMR under well-watered and drought conditions. Ten sugarcane genotypes (Uthong 6, Khon Kaen 80, K86-161, Khon Kaen 3, 03-4-425, KU60-1, Phill.66-07, B34-164, Uthong 2 and LF82-2122) and two water regimes (well-watered control and water stress at early growth stage) were laid out in factorial experiment in a randomized complete block design with two replications. Drought was imposed to the crop for 10 days during 90 days after transplanting (DAT) to 100 DAT. Data were recorded for total chlorophyll content by N, N-dimethylformamide extraction and SCMR on the second fully expanded leaf from the top at 90 DAT, 100 DAT and 110 DAT. Drought significantly reduced chlorophyll content and SCMR. The reduction in chlorophyll content was more severe than SCMR. Similar responses of sugarcane genotypes to drought and well-watered conditions were observed for chlorophyll content and SCMR. The breeding line 03-4-425 and the cultivars KU60-1 were identified as the best genotypes for chlorophyll content and SCMR. The correlation coefficients between chlorophyll content and SCMR were significant under well-watered conditions and the relationships were more strong and consistent under drought and drought relief. The SCMR can be used for evaluation of chlorophyll content under different water regimes.