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Enhancer trap systems have been demonstrated to increase the effectiveness of gene identification in rice. In this study, a  c hlorophyll- d eficient  m utant, named  oscdm1 , was screened and characterized in detail from a T-DNA enhancer-tagged population. The  oscdm1  plants were different from other chlorophyll-deficient mutants; they produced chlorotic leaves at the third leaf stage, which gradually died with further growth of the plants. However, the  oscdm1  plants were able to survive exposure to elevated CO 2  levels, similar to photorespiratory mutants. An analysis of the T-DNA flanking sequence in the  oscdm1  plants showed that the T-DNA was inserted into the promoter region of a serine hydroxymethyltransferase (SHMT) gene. OsSHMT1 is a key enzyme that is ubiquitous in nature and structurally conserved across kingdoms. The enzyme is responsible for the interconversion of serine and glycine and is essential for cellular one-carbon metabolism. Full-length  OsSHMT1  complemented the  oscdm1  phenotype, and the downregulation of  OsSHMT1  in wild-type plants by RNA interference (RNAi) produced plants that mimicked the  oscdm1  phenotype. GUS assays and quantitative PCR revealed the preferential expression of  OsSHMT1  in young leaves. TEM revealed serious damage to the thylakoid membrane in  oscdm1  chloroplasts. The  oscdm1  plants showed more extensive damage than wild type using an IMAGING-PAM fluorometer, especially under high light intensities. OsSHMT1-GFP localized exclusively to mitochondria. Further analysis revealed that the H 2 O 2  content in the  oscdm1  plants was twice that in wild type at the fourth leaf stage. This suggests that the thylakoid membrane damage observed in the  oscdm1  plants was caused by excessive H 2 O 2 . Interestingly,  OsSHMT1 -overexpressing plants exhibited increased photosynthetic efficiency and improved plant productivity. These results lay the foundation for further study of the  OsSHMT1  gene and will help illuminate the functional role of OsSHMT1 in photorespiration in rice.

The molecular cloning and clarification of a photorespiratory mutant, oscdm1, using enhancer trapping