基于HRM技术开发水稻糊化温度基因ALK功能标记
收稿日期: 2022-12-09
修回日期: 2023-02-16
网络出版日期: 2024-01-16
基金资助
江苏省重点研发计划项目(BE2020339);江苏省种业振兴揭榜挂帅项目(JBGS[2021]008)
Development of HRM-based Functional Marker for Gelatinization Temperature Gene ALK in Rice
Received date: 2022-12-09
Revised date: 2023-02-16
Online published: 2024-01-16
【目的】糊化温度是影响稻米蒸煮食味品质的重要指标，ALK是控制水稻糊化温度的主效基因，开发可快速高通量鉴定ALK基因型的功能标记有助于提高水稻品质改良的效率。【方法】根据ALK基因4198位、4329位和4330位存在的单碱基差异，设计基于HRM技术检测的基因功能标记。【结果】通过PCR检测结合测序分析，筛选了ALK基因2个功能区域的功能标记ALKH4、ALKH5。利用ALKH4、ALKH5对81份籼稻品种、279份粳稻品种进行了ALK基因型检测，结果发现，16份粳稻和19份籼稻为G-GC基因型；51份粳稻为A-GC基因型；212份粳稻和62份籼稻为G-TT基因型。【结论】基于HRM技术开发的基因功能标记ALKH4、ALKH5可以快速高通量鉴定控制糊化温度ALK不同基因型，具有重要的应用价值。
王军, 周晶, 陶亚军, 李文奇, 朱建平, 范方军, 王芳权, 许扬, 陈智慧, 蒋彦婕, 李霞, 杨杰 . 基于HRM技术开发水稻糊化温度基因ALK功能标记[J]. 中国水稻科学, 2024 , 38(1) : 106 -110 . DOI: 10.16819/j.1001-7216.2024.221203
【Objective】 Gelatinization temperature is an important parameter in rice eating quality which is mainly controlled by the ALK gene. Fast and high-throughput gene functional markers for ALK can improve selection efficiency in quality improvement. 【Method】 According to single nucleotide polymorphism in the functional region of ALK alleles, we developed the gene functional marker based on high resolution melting (HRM) technology. 【Result】 Through PCR amplification and sequencing test, we selected gene markers ALKH4 and ALKH5 in two functional regions of ALK. ALKH4 and ALKH5 were employed to genotype 81 indica varieties and 279 japonica varieties. The results showed that only 16 japonica varieties and 19 indica varieties carrying G-GC genotype were identified as well as 51 japonica varieties carrying A-GC genotype; while 212 japonica varieties and 62 indica varieties fell into G-TT genotype. 【Conclusion】 The HRM-based functional markers ALKH4 and ALKH5 could distinguish different genotypes of gelatinization temperature ALK locus in a fast and high-throughput manner, and have significance in breeding practice.
Key words： rice; gelatinization temperature; ALK; functional marker; HRM technology
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