In this study, family genes of ethylene biosynthesis (MAT, ACS and ACO) pathway in radish (Raphanus sativus L.) were genomewide identified by bioinformatics method and its tissuespecific response to biotic and abiotic stress were analyzed based on transcriptome data and quantitative realtime PCR. The results showed that: (1) a total of 8 MAT, 16 ACS and 7 ACO candidate genes were identified in radish genome and each family could be further divided into three subgroups. (2) The promoter region harbored at least one cisacting element that function in light responsiveness, and cisacting element involved in the phytohormones responsiveness were also found except for ACO1.1 and ACO3. The promoter region of some genes contains cisacting elements involved in response to biotic and abiotic stresses. (3) The analysis of transcriptome data showed that all MAT genes, ACS6.1 and ACO2/3/4 of radish have high expression levels in leaves. Agrobacterium tumefaciens infection significantly upregulated MAT24, ACS2/7.1/7.2 and ACO1.1/1.2/4 in resistance radish inbred lines, while most of them downregulated in susceptible material. The cadmium, chromium and lead stress significantly induced the expression of MAT4.2 and ACO1.1/4, while reduced the expression of MAT1 and ACO5.1. The expression of MAT2.1/2.2/4.1 and ACO1.1/5.2 were significantly suppressed by chilling stress of 4 ℃. (4) qRTPCR results indicated that both NaCl and PEG6000 stress significantly induced the expression of ACO5.1/5.2, but suppressed MAT4.1 and ACO4. Besides, exogenous fructose and sucrose may be involved in the response of radish to PEG6000 stress. Our findings in the present study will be helpful for the functional research of the ethylene biosynthesis genes in radish.