Choi J J, Alkharouf N W, Schneider K T, Matthews B F, Frederick R D
Foreign Disease-Weed Science Research Unit, USDA-Agricultural Research Service, 1301 Ditto Avenue, Fort Detrick, MD, 21702, USA.
Funct Integr Genomics. 2008 Nov;8(4):341-59. doi: 10.1007/s10142-008-0080-0. Epub 2008 Apr 15.
Soybean rust caused by Phakopsora pachyrhizi Sydow is a devastating foliar disease that has spread to most soybean growing regions throughout the world, including the USA. Four independent rust resistance genes, Rpp1-Rpp4, have been identified in soybean that recognize specific isolates of P. pachyrhizi. A suppressive subtraction hybridization (SSH) complementary DNA (cDNA) library was constructed from the soybean accession PI200492, which contains Rpp1, after inoculation with two different isolates of P. pachyrhizi that result in susceptible or immune reactions. Both forward and reverse SSH were performed using cDNA from messenger RNA pooled from 1, 6, 12, 24, and 48 h post-inoculation. A total of 1,728 SSH clones were sequenced and compared to sequences in GenBank for similarity. Microarray analyses were conducted on a custom 7883 soybean-cDNA clone array encompassing all of the soybean-rust SSH clones and expressed sequence tags from four other soybean cDNA libraries. Results of the microarray revealed 558 cDNA clones differentially expressed in the immune reaction. The majority of the upregulated cDNA clones fell into the functional category of defense. In particular, cDNA clones with similarity to peroxidases and lipoxygenases were prevalent. Downregulated cDNA clones included those with similarity to cell-wall-associated protein, such as extensins, proline-rich proteins, and xyloglucan endotransglycosylases.
由大豆锈菌(Phakopsora pachyrhizi Sydow)引起的大豆锈病是一种毁灭性的叶部病害,已蔓延至包括美国在内的全球大多数大豆种植区。在大豆中已鉴定出四个独立的抗锈病基因,即Rpp1 - Rpp4,它们可识别大豆锈菌的特定分离株。在接种导致感病或免疫反应的两种不同大豆锈菌分离株后,从含有Rpp1的大豆种质PI200492构建了一个抑制性消减杂交(SSH)互补DNA(cDNA)文库。正向和反向SSH均使用接种后1、6、12、24和48小时收集的信使RNA的cDNA进行。总共对1728个SSH克隆进行了测序,并与GenBank中的序列进行相似性比较。在一个包含所有大豆锈病SSH克隆以及来自其他四个大豆cDNA文库的表达序列标签的定制7883大豆cDNA克隆阵列上进行了微阵列分析。微阵列结果显示,在免疫反应中有558个cDNA克隆差异表达。大多数上调的cDNA克隆属于防御功能类别。特别是,与过氧化物酶和脂氧合酶相似的cDNA克隆很普遍。下调的cDNA克隆包括与细胞壁相关蛋白相似的那些,如伸展蛋白、富含脯氨酸的蛋白和木葡聚糖内转糖基酶。