Devey M E, Delfino-Mix A, Kinloch B B, Neale D B
Commonwealth Scientific and Industrial Research Organization/Division of Forestry, Australian Capital Territory, Australia.
Proc Natl Acad Sci U S A. 1995 Mar 14;92(6):2066-70. doi: 10.1073/pnas.92.6.2066.
We have genetically mapped a gene for resistance to white pine blister rust (Cronartium ribicola Fisch.) in sugar pine (Pinus lambertiana Dougl.) by using an approach which relies on three factors: (i) the ability to assay for genetic markers in the haploid stage of the host's life cycle, using megagametophyte seed tissue; (ii) a simple and clearly defined pathosystem; and (iii) the use of random amplified polymorphic DNA (RAPD) markers that can be quickly and efficiently evaluated. Resistance to white pine blister rust in sugar pine is known to be controlled by a single dominant gene (R). Maternal segregation of R and dominant RAPD markers were scored simultaneously following collection of megagametophytes for DNA assays and seedling inoculation with C. ribicola. Bulked samples of haploid megagametophyte DNA from resistant and susceptible offspring of segregating full-sib and half-sib families were used to evaluate 800 random decanucleotide primers. Ten loci linked with the gene for resistance to white pine blister rust were identified and segregation data were obtained from five families. Six of the linked markers were within 5 centimorgans of the gene, and one marker was 0.9 centimorgan from R. These and other markers derived by this approach may provide starting points for map-based cloning of this important gene.
我们通过一种依赖于三个因素的方法,对糖松(Pinus lambertiana Dougl.)中抗白松疱锈病(Cronartium ribicola Fisch.)的基因进行了遗传定位:(i)利用大配子体种子组织,在寄主生命周期的单倍体阶段检测遗传标记的能力;(ii)一个简单且明确界定的病理系统;(iii)使用能够快速且高效评估的随机扩增多态性DNA(RAPD)标记。已知糖松对白松疱锈病的抗性由单个显性基因(R)控制。在收集大配子体用于DNA检测以及用C. ribicola对幼苗进行接种后,同时对R和显性RAPD标记的母本分离情况进行评分。来自分离的全同胞和半同胞家系的抗性和感病后代的单倍体大配子体DNA混合样本,用于评估800个随机十聚体引物。鉴定出了10个与抗白松疱锈病基因连锁的位点,并从5个家系获得了分离数据。其中6个连锁标记位于该基因的5厘摩范围内,一个标记与R的距离为0.9厘摩。通过这种方法获得的这些及其他标记可为该重要基因的图位克隆提供起点。