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首页> 外文会议>International Rubus and Ribes Symposium >Developing Molecular Markers for Marker Assisted Selection for Resistance to Raspberry Bushy Dwarf Virus (RBDV) in Red Raspberry
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Developing Molecular Markers for Marker Assisted Selection for Resistance to Raspberry Bushy Dwarf Virus (RBDV) in Red Raspberry

机译：开发用于标记物辅助选择的分子标记对红覆盆子抗覆盆子浓密矮化病毒（RBDV）的抗性

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A modified Bulked Segregant Analysis (BSA) was applied using eight RBDV resistant and eight susceptible raspberry plants. The best markers from this effort were applied to sixty seedlings of a 'Nootka' x WSU1499 population segregating for resistance to RBDV conferred by the Bu resistance gene. ELISA was used to identify resistant and susceptible genotypes in this population following graft inoculations. Three markers showed promise for use in Marker Assisted Selection (MAS); BC615_550, BC002_590 and BC296_425. BC615_550 proved to be the most predictive and reproducible marker and was cloned and sequenced. Specific primers were developed for this marker, resulting in the BC615_553_AluI Cleaved Amplified Polymorphic Sequence (CAPS) marker. This marker was applied to the segregating population and correctly identified the resistant plants in 58 out of 60 cases (96.7% accuracy). This marker was not present in other notable resistant sources, such as that derived from 'Haida' so a comparative approach based on assumed microsynteny between raspberry and strawberry was implemented. A BLAST search identified the putative location of the BC615_550 polymorphism in the diploid strawberry genome and primers were designed in candidate genes physically linked to this locus. Of note, six primer sets were designed to amplify different parts of a gene containing motifs homologous to the tobacco N-gene, a known virus resistance gene. One of the primer sets resulted in a Sequence Characterized Amplified Region (SCAR) marker, called rasp_N_gene_1202, that amplified in the resistant plants 'Nootka', 'Haida', and 'Willamette' in initial tests, while failing to amplify in the susceptible WSU1499. In the test population, this marker segregates perfectly with the original BC615_553_AluI marker. In addition to 'Nootka', 'Haida', and 'Willamette', it is present in a diverse set of known resistant genotypes including WSU 78117-1, 'Newburgh', 'Cowichan', 'Chilcotin', 'Malling Promise', 'Heritage' and 'Latham'. The marker is absent in known susceptible genotypes including 'Chilliwack', 'Tulameen', WSU 1276, 'Meeker', 'Autumn Bliss', 'Malahat' and WSU 1098. Screening of additional test populations is in progress for the evaluation of the marker's utility in MAS for RBDV resistance in red raspberry.
机译：使用八个RBDV抗性和八个敏感的覆盆子植物施用改性的致磨的偏析分析（BSA）。来自这种努力的最佳标记应用于“NOOTKA”X WSU1499群体的六十幼苗，用于赋予Bu抗性基因的RBDV的抵抗力。在接枝接收后，ELISA用于鉴定该群体中的抗性和敏感基因型。三个标记显示在标记辅助选择（MAS）中使用的承诺; BC615_550，BC002_590和BC296_425。 BC615_550被证明是最具预测性和可重复的标记，并被克隆和测序。为该标记开发了特异性引物，得到了BC615_553_ALUI切割的扩增多态序列（盖子）标记。将该标记物应用于隔离群体，并正确鉴定了60例中的58例（精度为96.7％）中的58种抗性植物。该标记不存在于其他显着的抗性源中，例如来自'Haida'，因此实施了基于覆盆子和草莓之间的假定微生物的比较方法。 Blast搜索确定了二倍体草莓基因组中BC615_550多态性的推定位置，并设计了与该基因座物理相关的候选基因中的引物。值得注意的是，设计六个引物套以扩增含有与烟草N-基因同源的基因的基因的不同部分，该基因是已知的病毒抗性基因。其中一个引物组导致序列表征的扩增区域（瘢痕）标记，称为RASP_N_GENE_1202，其在初始测试中，在耐药设备'NOOTKA'，'HAIDA'和“WILLAMETTE”中放大，同时在易感WSU1499中无法放大。在测试人群中，这个标记与原始BC615_553_ALUI标记完全分离。除了'nootka'，'haida'和'willamette'之外，它是一种不同的已知抗性基因型，包括Wsu 78117-1，'纽伯格'，'辣椒'，'chilcotin'，'Malling Promise'， 'heritage'和'latham'。在众所周知的易感基因型中，包括'Chilliwack'，'tulameen'，WSU 1276，'Meeker'，'秋季幸福'，'Malahat'和WSU 1098中的标记。筛选额外的测试人口正在评估标记的评估红覆盆子中抗RBDV抗性的MAS效用。

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