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以高抗穗枯病野生车前草种质PPZ14为材料,用引起车前草穗枯病的当归间座壳菌Diaporthe angelicae强致病力真菌菌株DA‐8对其接种,采用cDNA‐AFLP技术分离接种后不同时间点产生的差异表达基因片段。结果从64对引物组合中筛选到4对多态性丰富的引物组合,并用其进行扩增得到202条差异表达的转录衍生片段(transcript‐derived fragment,TDF),其中以接种24h后出现的TDF数量最多,表明在此时间点车前草与当归间座壳菌互作最活跃。对部分TDF进行克隆及序列测定,共获得7个TDF的核苷酸序列和推导的氨基酸序列,经BLAST比对,5个TDF的氨基酸序列在GenBank中找到与其对应的显著同源序列,其中4个为已知功能的基因,对其进行功能分析后,推测认为,决定ACC氧化酶、细胞色素C生成蛋白C、液泡膜H+‐ATPase的TDF与车前草抗穗枯病性相关。
In this study, PPZ14 was used to inoculate the wild plantago psyllium PPZ14, which is highly resistant to head-blight, and inoculated with the virulent strain DA-8 of Diaporthe angelicae, Differentially expressed gene fragments were generated at different time points after isolation. Results Four pairs of primer combinations were screened from 64 pairs of primers and amplified by PCR. Totally 202 differentially expressed transcript-derived fragments (TDFs) were obtained. TDF The largest number, indicating that at this time point between plantago and Angelica aculeatus most active interaction. A total of seven TDF nucleotide sequences and deduced amino acid sequences were obtained by cloning and sequencing of partial TDFs. By BLAST analysis, five TDF amino acid sequences were found to have significant homologous sequences in GenBank, of which 4 After analyzing the function of these genes, we hypothesized that TDF, which determines ACC oxidase, cytochrome C-producing protein C, and tonoplast H + -ATPase, is associated with resistance to panicle disease.