贺玉花 户克云 赵光伟 孔维虎 张健 徐永阳

摘    要：纯度是决定种子质量的重要指标之一。以众天10号和其父母本为研究材料，将快提DNA法与多重PCR法相结合，成功建立一套三重PCR甜瓜杂种纯度快速鉴定体系。采用碱裂解快速提取DNA的方法，可以在5 min内提取96个DNA;采用三重PCR，能有效克服单一PCR在种子纯度鉴定中速率慢的缺陷。试验中SSR分子鉴定众天10号杂交种纯度为100%，与田间鉴定结果一致。为甜瓜乃至作物杂种种子的纯度鉴定提供了一套精度更高、速度更快、成本更低的技术体系。

关键词：甜瓜;SSR;多重PCR;快提DNA;纯度鉴定

中图分类号：S652 文献标志码：A 文章编号：1673-2871（2021）12-023-05

Abstract： Purity is one of the most important indicators of seed quality. A hybrid Zhongtian No. 10 and their parents were used in this study; and by combining the rapid DNA extraction method with the multiple PCR， a triple PCR purity identification system of hybrid seeds in melon was established. The rapid alkaline lysis method was used to extract the genome DNA， which can extract the genomic DNA of 96 samples in five minutes. The triplex PCR could effectively overcome the shortcomings of the single-PCR with slow speed in seed purity identification. The result of SSR molecular identification was that the purity of the Zhongtian No. 10 hybrid was 100%， and it was no significant difference with field identification. This study provides a technical system with the advantages of high-accuracy， rapid and low-cost for the purity identification of hybrid seeds in melon and other crop.

Key words： Melon; SSR; Multiplex PCR; Rapid DNA extraction; Purity identification

甜瓜（Cucumis melo L.）又名蜜瓜或香瓜，原产于印度、非洲热带沙漠地区[1-2]，是喜温、耐热性较强的一年生蔓生作物。中国甜瓜面积占世界总面积的45%以上，产量占55%以上[3]，是我国重要的经济水果之一。市面上销售的甜瓜种子大部分为杂交1代，而种子纯度是生产和销售过程中重要的指标之一。目前，种子纯度的检测方法主要采用传统的田间形态学方法，周期较长且花费较高，受自然环境条件影响大。分子标记鉴定技术具有周期短、准确度高等优点[4]。简单重复序列（simple sequence repeats，SSR）是目前最常用的纯度鉴定标记之一，具有多态性、共显性和高度重复性等优点，被广泛应用于许多作物的杂种纯度鉴定[4-13]。

在利用SSR分子标记鉴定杂种纯度的过程中，DNA提取和PCR扩增是关键的步骤。传统的DNA提取方法试验操作复杂，费时费力，改良后的碱裂解快提DNA方法可在5 min提取96个DNA，大大缩短鉴定周期[14]。……