Welcome to Chinese Agricultural Science Bulletin,

Chinese Agricultural Science Bulletin ›› 2020, Vol. 36 ›› Issue (6): 69-73.doi: 10.11924/j.issn.1000-6850.casb18100116

Special Issue: 生物技术 小麦

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Comparison of DNA Extraction of Different Tartary Buckwheat Parts After Drying and Screening of SSR Primers Related to Thin Shell Characters

Yin Guifang, Li Chunhua, Sun Daowang, Lu Wenjie, Wang Yanqing, Wang Lihua()   

  1. Biotechnology and Germplasm Resources Institute, Yunnan Academy of Agricultural Sciences/Yunnan Provincial Key Lab of Agricultural Biotechnology/Key Lab of Southwestern Crop Gene Resources and Germplasm Innovation, Ministry of Agriculture, Kunming 650205
  • Received:2018-10-30 Revised:2018-11-28 Online:2020-02-25 Published:2020-02-22
  • Contact: Wang Lihua E-mail:wanglihua70@hotmail.com

Abstract:

The study aims to provide a basis for expanding the range of DNA extraction materials of tartary buckwheat, and to screen thin shell traits-related SSR primers, thus to lay a foundation for molecular markers of specific traits. Six parts of tartary buckwheat plants were used as materials for genomic DNA extraction, including cotyledons, tender stems, tender leaves, old leaves, old stems and petioles, and genomic DNA extraction was carried out using the plant DNA extraction kit after the materials were dried in an oven at 40℃. The quality, concentration and purity of DNA were detected. A total of 700 pairs of SSR primers were used for PCR amplification to screen thin shell traits-related SSR primers of tartary buckwheat. The results showed that the highest DNA concentration was obtained from the cotyledons (70.6 ng/μL), followed by tender stems (69.6 ng/μL); and lower NDA concentrations were obtained from the old stems and the petioles, which were 20.0 ng/μL and 7.2 ng/μL, respectively. The gel electrophoresis bands of DNA extracted from the cotyledons, the tender stems, the tender leaves and the old leaves were clear, while gel electrophoresis bands of DNA extracted from the old stems and the petioles were dark. Regarding the SSR amplification effect, except petioles, the cotyledons, the tender stems, the tender leaves, the old leaves and the old stems could meet the requirements of amplification without significant difference, and they could be used in subsequent molecular experiments. The DNA concentrations from dried tissues were not as high as those from fresh tissues, but the subsequent molecular experiments were not affected, except petioles. A pair of SSR primers with diversity was preliminarily screened in thin-shell tartary buckwheat and thick-shell tartary buckwheat.

Key words: tartary buckwheat, drying, DNA extraction, SSR

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