College of Plant Protection and Shaanxi Key Laboratory of Molecular Biology for Agriculture, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
BMC Plant Biol. 2010 Jan 12;10:9. doi: 10.1186/1471-2229-10-9.
Stripe rust of wheat, caused by Puccinia striiformis f. sp. tritici (Pst), is one of the most important diseases of wheat worldwide. Due to special features of hexaploid wheat with large and complex genome and difficulties for transformation, and of Pst without sexual reproduction and hard to culture on media, the use of most genetic and molecular techniques in studying genes involved in the wheat-Pst interactions has been largely limited. The objective of this study was to identify transcriptionally regulated genes during an incompatible interaction between wheat and Pst using cDNA-AFLP technique
A total of 52,992 transcript derived fragments (TDFs) were generated with 64 primer pairs and 2,437 (4.6%) of them displayed altered expression patterns after inoculation with 1,787 up-regulated and 650 down-regulated. We obtained reliable sequences (>100 bp) for 255 selected TDFs, of which 113 (44.3%) had putative functions identified. A large group (17.6%) of these genes shared high homology with genes involved in metabolism and photosynthesis; 13.8% to genes with functions related to disease defense and signal transduction; and those in the remaining groups (12.9%) to genes involved in transcription, transport processes, protein metabolism, and cell structure, respectively. Through comparing TDFs identified in the present study for incompatible interaction and those identified in the previous study for compatible interactions, 161 TDFs were shared by both interactions, 94 were expressed specifically in the incompatible interaction, of which the specificity of 43 selected transcripts were determined using quantitative real-time polymerase chain reaction (qRT-PCR). Based on the analyses of homology to genes known to play a role in defense, signal transduction and protein metabolism, 20 TDFs were chosen and their expression patterns revealed by the cDNA-AFLP technique were confirmed using the qRT-PCR analysis.
We uncovered a number of new candidate genes possibly involved in the interactions of wheat and Pst, of which 11 TDFs expressed specifically in the incompatible interaction. Resistance to stripe rust in wheat cv. Suwon11 is executed after penetration has occurred. Moreover, we also found that plant responses in compatible and incompatible interactions are qualitatively similar but quantitatively different soon after stripe rust fungus infection.
小麦条锈病是由条形柄锈菌小麦专化型(Pst)引起的,是世界范围内小麦最重要的病害之一。由于六倍体小麦基因组庞大且复杂,转化困难,而 Pst 没有有性繁殖,难以在培养基上培养,因此,在研究与小麦- Pst 相互作用相关的基因时,大多数遗传和分子技术的应用受到了很大限制。本研究的目的是使用 cDNA-AFLP 技术鉴定小麦与 Pst 不亲和互作过程中转录调控基因。
利用 64 对引物共产生了 52992 条转录物衍生片段(TDFs),其中 1787 条上调,650 条下调,2437 条(4.6%)表达模式发生改变。对 255 个选定的 TDFs 进行了可靠的序列(>100bp)分析,其中 113 个(44.3%)具有已鉴定功能。这些基因的一个大组(17.6%)与参与代谢和光合作用的基因具有高度同源性;13.8%与与疾病防御和信号转导相关的功能基因有关;其余各组(12.9%)分别与转录、运输过程、蛋白质代谢和细胞结构有关。通过比较本研究中用于不亲和互作鉴定的 TDFs 和之前用于亲和互作鉴定的 TDFs,发现 161 个 TDFs在两种互作中都有表达,94 个在不亲和互作中特异性表达,其中 43 个选定转录物的特异性通过定量实时聚合酶链反应(qRT-PCR)确定。根据与已知在防御、信号转导和蛋白质代谢中发挥作用的基因的同源性分析,选择了 20 个 TDFs,并通过 qRT-PCR 分析证实了它们在 cDNA-AFLP 技术中的表达模式。
本研究揭示了一批可能参与小麦与 Pst 相互作用的新候选基因,其中 11 个 TDFs在不亲和互作中特异性表达。小麦品种 Suwon11 对条锈病的抗性是在穿透发生后执行的。此外,我们还发现,在条锈菌感染后,亲和和不亲和互作的植物反应在定性上相似,但在定量上不同。