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| Purity Identification and SNP Site Obtain of Chinese Cabbage Hybrids Using
Multiplex EST-SSR Marker |
| ZHAO Xin1，WANG Yong1*，LAN Qing-kuo1，HE Chang-zheng2，CHEN Rui1，LI Ou-jing1，LIU
Na1，ZHU Zhu1，GUO Yong-ze1 |
（1Institute of Tianjin Agriculture Quality Standard and Testing Technology， Tianjin 300381， China； 2Tianjin Seed
Station， Tianjin 300061， China） |
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Abstract In this research，10 Chinese cabbage〔Brassica campestris L. ssp. pekinensis（Lour）
Olsson〕hybrids and their parents were taken as study objects．Based on the EST sequences from
NCBI genome data base，a SNP site and multiplex EST- SSR sites，which could be used to identify the
purity of Chinese cabbage hybrids，were screened by SSR marker and high resolution melting（HRM）
technology．According to the requirement of seed purity identification by high throughput molecular
technology，the key techniques such as seedling culture condition of single seed，extraction status of
single grain plant, rapid DNA extraction method and establishment of multiplex PCR system were groped
and optimized．Results showed that the single germinating seed after 48 h could gain DNA with higher
quality by the improved CTAB method in 3 h. Meanwhile，plant leaves reached seeding stage after 7
d，could complete DNA extraction by chexe-100 method within 40 min．The SNP site obtained by screening could be used to indentify the purity of 6 Chinese cabbage hybrids，and the multiplex ESTSSR
locus could be used to indentify the purity of 10 Chinese c abbage hybrids．
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Received: 25 March 2013
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