Johnston P. A., Pickering R. A.
New Zealand Institute for Crop and Food Research Limited, Private Bag 4704, Christchurch, New Zealand.
Theor Appl Genet. 2002 Mar;104(4):720-726. doi: 10.1007/s00122-001-0791-2.
Retrotransposon-like sequences are ideal tools for initial screening assays to distinguish between closely related species because of their ubiquitous presence, high copy number, chromosome coverage and rapid sequence evolution. A retrotransposon-like sequence, pSc119.1, cloned from Secale cereale (rye) has been used to obtain PCR primers that are capable of detecting small introgressions of Hordeum bulbosum (bulbous barley grass) chromatin in a Hordeum vulgare (cultivated barley) background. Combining this PCR-based assay with a crude but effective high-throughput DNA extraction has enabled the rapid identification of plants possessing H. bulbosumintrogressions from large numbers of progeny from H. vulgarex H. bulbosumcrosses. These plants are then further characterized by more-refined cytological, molecular and pathological techniques to locate and map the introgressed chromatin and to evaluate their disease resistance.
由于反转录转座子样序列普遍存在、拷贝数高、染色体覆盖范围广且序列进化迅速,因此它们是区分近缘物种的初始筛选试验的理想工具。从黑麦(Secale cereale)克隆的一个反转录转座子样序列pSc119.1已被用于获得能够检测在栽培大麦(Hordeum vulgare)背景下球茎大麦(Hordeum bulbosum)染色质小渗入片段的PCR引物。将这种基于PCR的检测方法与一种粗糙但有效的高通量DNA提取方法相结合,能够从大量大麦与球茎大麦杂交后代中快速鉴定出具有球茎大麦渗入片段的植株。然后,通过更精细的细胞学、分子和病理学技术对这些植株进行进一步鉴定,以定位和绘制渗入染色质图谱,并评估它们的抗病性。