【作者】 陈官菊；

【导师】 陈利锋；

【作者基本信息】 南京农业大学 ， 植物病理学， 2005， 硕士

【摘要】 本研究分为两部分:1.运用RT-PCR技术对禾谷镰孢与小麦寄主互作过程中毒素相关基因的表达情况进行了分析;2.运用PCR技术对我国禾谷镰孢产毒种类进行了检测。 采用RT-PCR技术,辅以生物分析软件(Bio-Rad Quantity One)对禾谷镰孢与小麦互作过程中毒素相关基因Tri5、Tri6、Tri10以及Zri12的表达量进行了研究。结果显示,在24hai(hours after inoculation),Tri5在苏麦3号和宁麦6号上的相对表达量无显著差异,但在36～72hai,Tri5在感病品种宁麦6号上的相对表达量为抗病品种苏麦3号上的2倍左右。Tri6和Tri10的相对表达量在抗、感品种上没有显著差异。而在48hai和72hai,Tri12的相对表达量在宁麦6号和苏麦3号上差异显著,前者分别为后者的4.4和4.9倍,由此我们推测,苏麦3号可能存在抑制和延缓毒素产生及阻止毒素转运的抗病机制。 运用两组(各4个)能特异性检测分别产生NIV、DON与3-ADON、DON与15-ADON的禾谷镰孢菌株的引物,对来自我国13个省份的316个禾谷镰孢菌株进行了产毒种类的检测。结果表明,在供试的所有菌株中,有49个菌株产NIV,占供试菌株的15.5%;136个菌株产DON和15-ADON,占供试菌株的43.0%;131个菌株产DON和3-ADON,占供试菌株的41.5%。产DON的菌株占绝对优势。其中在江苏和安徽未检测到产15-ADON的菌株;在福建和浙江两省未检测到产3-ADON的菌株;在湖北和河南两省未检测到产NIV的菌株。河北省、宁夏回族自治区和山东省的被检菌株只产DON和15-ADON;江西省的被检菌株只产DON和3-ADON;四川省的被检菌株只产NIV。在黑龙江和贵州两省的菌株中可产生上述4种毒素。所有禾谷镰孢的产毒种类与系群的归属和菌株来源小麦品种的关系不明显。更多还原

【Abstract】 Trichothecenes are a group of secondary metabolites in Fusarium. Tri5, Tri6, Tri10 and Tri12 are among the 16 trichothecene biosynthesis genes reported so far. In this study, we analyzed the expression level of Tri5、, Tri6、 TrilO and Tri12 during the interaction between Fusarium graminearum and wheat by using RT-PCR technology and Molecular Analyst software (Bio-Rad Quantity One). We observed that there is no detectable level of expression of the four genes at 0—12 hai (hours after inoculation). At 24 hai, the relative expression level of Tri5 in resistant cultivar Sumai 3 and susceptible cultivar Ningmai 6 had no significant difference. But at 36~72 hai, the expression level of Tri5 in Ningmai 6 was about twice of that in Sumai 3. The relative expression of Tri6 and TrilO showed no significant difference at all time points whether it was on resistant or susceptible cultivar. At 48 hai and 72 hai, the relative expression of Tril2 in Ningmai 6and Sumai 3 showed significant difference, the former being 4.4 times and 4.9 times of the latter. Thus we hypothesized that Sumai 3 may not only degrade trichothecenes, but also inhibit the transport of trichothecenes as well.We also collected wheat heads showing symptoms of Fusarium head blight from 13 provinces of China. A total of 316 isolates of Fusarium graminearum were subjected to differentiation of the isolates that produce NIV-, DON, 3-ADON, and 15-ADON, respectively, using PCR with two sets (four of each) of primers. The results showed that 49 isolates (15.5%) produced NIV, 136 isolates (43.0%) produced DON and 15-ADON, 131 isolates (41.5%) produced DON and 3-ADON. In summary, most isolates produce DON. Among those isolates, the 15-ADON-producing isolates were not found in Jiangsu Province and Anhui Province, the 3-ADON-producing isolates were not found in Fujian Province and Zhejiang Province, the NIV-producing isolates were not found in Hubei Province and Henan Province. The isolates from Hebei Province, Ningxia Province and Shandong Province only produce DON and 15-ADON, while the isolates from Jiangxi Province only produce DON and 3-ADON, and the isolates from Sichuan Province only produce NIV. The isolates from Heilongjiang Province and Guizhou Province canproduce all four tested toxins. No significant correlation between Fusarium graminearum toxin-producing ability and their lineage designation and wheat cultivars from which they were isolated from was shown.更多还原