Abstract: Targeted editing the rice（Oryza sativa） thylakoid lumen protein gene OsTLP16.5（Os06g0705100） using CRISPR/Cas9technology,most of the amino acid sequences encoded by the gene were changed,and the mutant ostlp16.5 was screened and identified.Metabonomic analysis of rice mutant ostlp16.5 and wild type ZH11 showed that the contents of organic acids,amino acids,vitamins and their derivatives were significantly different between mutant ostlp16.5 and wild type ZH11.Further analysis showed that the metabolites（3,4-dihydroxyphenylpropanoate,dopamine,L-arogenate acid and L-tyrosine,L-2-hydroxyglutaric acid,2-oxoarginine,pantothenic acid,etc.） and the metabolic pathways（isoquinoline alkaloid biosynthesis,tyrosine metabolism,etc.） with significant differences between mutant ostlp16.5 and wild type ZH11 were mostly directly or indirectly related to photosynthetic reaction.These results indicated that OsTLP16.5 protein may affect the metabolism of organic acids,amino acids and vitamins in rice by participating in photosynthetic regulation.The results are important for revealing the function of OsTLP16.5 and improving the productivity for rice,and could provide a reference for studying OsTLP16.5 protein regulation of secondary metabolism and growth and development in rice.