Discovery of a nuclear haplotype potentially useful for the identification of medicinal rice Njavara (Oryza sativa L.)

R. Raj, U. Suresh Kumar, M. R. Vinitha, B. C. Patra, O. N. Singh, George Thomas
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Abstract

The present study report the development of an amplified fragment length polymorphism (AFLP)-derived sequence characterized amplified region (SCAR) marker for reliable identification of Njavara (Oryza sativa landrace Njavara), a rice landrace in India used extensively in health foods and Ayurveda treatments. The SCAR locus, named OsNSCAR131 after O. sativa landrace Njavara, is located on chromosome 3 between nucleotide positions 7793982 and 7794108, and yields a 131-bp allele in Njavara accessions and a 127-bp allele in other rice cultivars due to a 4-bp Insertion-Deletion (InDel) mutation at nucleotide position 7794026. We standardized the SCAR assay to be like those used for detecting microsatellite markers by using fluorescently (6-FAM) labeled primers and separating the alleles by capillary electrophoresis. As an alternative, we further adapted the method so as to allow allele detection by polyacrylamide gel electrophoresis (PAGE). A single rice grain can be tested for authentication by this PAGE assay. The SCAR marker developed here has great utility in authenticating Njavara grains in both the health food and pharmaceutical sectors.
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发现可用于药用水稻 Njavara(Oryza sativa L.)鉴定的核单倍型
本研究报告开发了一种由扩增片段长度多态性(AFLP)衍生的序列特征扩增区域(SCAR)标记,用于可靠鉴定印度水稻陆稻品种 Njavara(Oryza sativa landrace Njavara),该品种被广泛用于保健食品和阿育吠陀疗法。SCAR 位点以 O. sativa 陆稻品种 Njavara 命名为 OsNSCAR131,位于 3 号染色体核苷酸位置 7793982 和 7794108 之间,在 Njavara 品种中产生一个 131-bp 的等位基因,在其他水稻栽培品种中产生一个 127-bp 的等位基因,原因是核苷酸位置 7794026 存在一个 4-bp 的插入-缺失(InDel)突变。通过使用荧光(6-FAM)标记引物和毛细管电泳分离等位基因,我们将 SCAR 检测方法标准化,使其与检测微卫星标记的方法相同。作为一种替代方法,我们进一步调整了该方法,以便通过聚丙烯酰胺凝胶电泳(PAGE)检测等位基因。通过这种聚丙烯酰胺凝胶电泳检测法,可以对一粒稻谷进行鉴定。这里开发的 SCAR 标记在保健食品和制药行业鉴定 Njavara 谷物方面都有很大的用途。
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