苦荞麦不同部位干燥后DNA提取效果的比较及薄壳性状关联SSR引物筛选研究

旨在为扩大苦荞麦DNA提取材料范围提供依据,筛选与薄壳性状相关联SSR引物,为苦荞麦特异性状分子标记奠定基础。分别以苦荞麦植株的子叶、嫩茎、嫩叶、老叶、老茎、叶柄为材料,40℃烘箱烘干后,采用植物DNA提取试剂盒,分别提取苦荞麦6个部位的基因组DNA,并进行DNA质量、浓度和纯度检测,利用700对SSR引物进行PCR扩增,筛选与苦荞麦薄壳性状相关联引物。结果表明：子叶提取的DNA浓度最高,为70.6 ng/μL;嫩茎次之,为69.6 ng/μL;老茎和叶柄获得的NDA浓度偏低,分别为20.0 ng/μL和7.2 ng/μL。采用子叶、嫩茎、嫩叶、老叶提取的DNA凝胶电泳条带清晰,采用老茎和叶柄提取的DNA凝胶电泳条带暗。SSR扩增效果除叶柄不能达到扩增要求外,其他没有明显差异都能达到扩增要求,可用于后续的分子实验。采用烘干组织提取DNA浓度虽然没有新鲜组织高,但除叶柄外都不影响后续分子试验。初步筛选出在薄壳和厚壳苦荞麦中具有多样性的SSR引物1对。

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

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.