中文摘要：

根据橡胶树白粉菌（Oidium heveae Steinm.）基因组中的特有保守序列OHS,设计两对特异性引物OHF1/OHR1和OHF2/OHR2。以不同地区收集的6份O.heveae（OH1~OH6）和橡胶树胶孢炭疽病菌（Colletotrichum gloeosporioides Penz.）等5种非靶标病原菌及健康橡胶树叶片基因组DNA为模板,建立了橡胶树白粉菌PCR及nested-PCR分子检测技术,并验证了检测体系的特异性和灵敏度。结果表明,引物OHF1/OHR1和OHF2/OHR2对橡胶树白粉菌均具有较高的特异性和灵敏度。其中引物OHF2/OHR2能检测到10pg/μL的橡胶树白粉菌DNA,而以OHF1/OHR1和OHF2/OHR2组合进行nested-PCR,其最低DNA检测浓度达到0.01fg/μL。人工接种试验中,当孢子接种量为2×10~3个/叶时,PCR和nested-PCR检测体系可分别在接种4d和24h后检测到目的条带。表明nested-PCR对在叶片组织中处于潜育期的橡胶树白粉菌的检测更有效,可为橡胶树白粉菌的检测提供一种简便而准确的检测方法。

英文摘要：

To establish methods for detection of powdery mildew of rubber tree by using conventional PCR and nested- PCR, two sets of specific primers OHF1/OHR1 and OHF2/OHR2 were designed respectively based on unique conserved sequence OHS of Oidiurn heveae. Six O. heveae samples （OH1-OH6） from different region, five strains of non-target pathogenic fungi and healthy rubber tree leaves were used to evaluate the specificity and sensitivity of conventional PCR and nested-PCR detection system. The results revealed that both OHF1/OHR1 and OHF2/OHR2 have high specificity and sensitivity. The detection limit is 10 pg/μL and 0.01 fg/μL DNA, respectively for conventional PCR （using primer OHF2/OHR2） and nested-PCR （using OHF1/OHR1 and OHF2/OHR2）. Artificial inoculation test of O. heveae on rubber tree indicated that target band can be detected by PCR and nested-PCR at 4th day and 24 h after inoculation, respectively. These results suggested that the nested-PCR is more efficient than the conventional PCR for O. heveae in incubation period and provide an easy and accurate way for O. heveae detection.