International Crops Research Institute for the Semi-Arid Tropics, Patancheru, India.
Funct Integr Genomics. 2011 Dec;11(4):651-7. doi: 10.1007/s10142-011-0227-2. Epub 2011 May 6.
Single feature polymorphisms (SFPs) are microarray-based molecular markers that are detected by hybridization of DNA or cRNA to oligonucleotide probes. With an objective to identify the potential polymorphic markers for drought tolerance in pigeonpea [Cajanus cajan (L.) Millspaugh], an important legume crop for the semi-arid tropics but deficient in genomic resources, Affymetrix Genome Arrays of soybean (Glycine max), a closely related species of pigeonpea were used on cRNA of six parental genotypes of three mapping populations of pigeonpea segregating for agronomic traits like drought tolerance and pod borer (Helicoverpa armigiera) resistance. By using robustified projection pursuit method on 15 pair-wise comparisons for the six parental genotypes, 5,692 SFPs were identified. Number of SFPs varied from 780 (ICPL 8755 × ICPL 227) to 854 (ICPL 151 × ICPL 87) per parental combination of the mapping populations. Randomly selected 179 SFPs were used for validation by Sanger sequencing and good quality sequence data were obtained for 99 genes of which 75 genes showed sequence polymorphisms. While associating the sequence polymorphisms with SFPs detected, true positives were observed for 52.6% SFPs detected. In terms of parental combinations of the mapping populations, occurrence of true positives was 34.48% for ICPL 151 × ICPL 87, 41.86% for ICPL 8755 × ICPL 227, and 81.58% for ICP 28 × ICPW 94. In addition, a set of 139 candidate genes that may be associated with drought tolerance has been identified based on gene ontology analysis of the homologous pigeonpea genes to the soybean genes that detected SFPs between the parents of the mapping populations segregating for drought tolerance.
单特征多态性(SFPs)是基于微阵列的分子标记,通过 DNA 或 cRNA 与寡核苷酸探针杂交来检测。为了鉴定半干旱热带地区重要的豆科作物绿豆(Cajanus cajan(L.)Millspaugh)对干旱的潜在多态性标记,利用与绿豆密切相关的大豆(Glycine max)的 Affymetrix 基因组阵列对三个遗传图谱群体的六个亲本基因型的 cRNA 进行了研究,这些群体分离出了对干旱和豆荚螟(Helicoverpa armigera)抗性等农艺性状的抗性。通过对六个亲本基因型的 15 对比较使用稳健化投影追踪方法,鉴定出 5692 个 SFPs。每个图谱群体的亲本组合的 SFPs 数量从 780(ICPL 8755×ICPL 227)到 854(ICPL 151×ICPL 87)不等。随机选择了 179 个 SFPs 进行 Sanger 测序验证,获得了 99 个基因的高质量序列数据,其中 75 个基因显示出序列多态性。在将序列多态性与检测到的 SFPs 相关联时,检测到的 52.6% SFPs 观察到真正的阳性结果。就图谱群体的亲本组合而言,ICPL 151×ICPL 87 的阳性结果发生率为 34.48%,ICPL 8755×ICPL 227 为 41.86%,ICPL 28×ICPW 94 为 81.58%。此外,基于对在分离耐旱性的图谱群体亲本之间检测到的 SFPs 的大豆基因与同源绿豆基因的基因本体分析,已经确定了一组可能与耐旱性相关的 139 个候选基因。