Broom's Barn Research Station, Higham, Bury St Edmunds, Suffolk, IP28 6NP, UK.
Mol Plant Pathol. 2003 May 1;4(3):171-6. doi: 10.1046/j.1364-3703.2003.00160.x.
SUMMARY We have tested and developed protocols for both sequence-independent and hybridization probe real-time PCR for the detection of Polymyxa betae glutathione-S-transferase transcripts in infected sugar beet roots. When using the test on P. betae-free plants, no signal above the level of the non-template control was observed. Real-time PCR analysis of both serially diluted zoospore suspensions and infected root material demonstrated a close relationship between the threshold cycle and the amount of P. betae. Hybridization probe real-time analysis of infected plants sampled sequentially over 20 days from sowing showed that the levels of the transcript rose steadily after initial infection to a peak and then declined. Comparative time-course analyses of infection in susceptible plants and a resistant wild Beta species indicated that, whilst transcript levels in susceptible plants showed a continuing upward trend, in the resistant species they were detectable only at an extremely low level.
摘要 我们已经测试和开发了用于序列无关和杂交探针实时 PCR 的方案，以检测感染甜菜根中的多粘类芽孢杆菌谷胱甘肽 S-转移酶转录物。当在不含 P. betae 的植物上使用该检测方法时，在非模板对照水平之上没有观察到信号。对连续稀释的游动孢子悬浮液和感染根材料的实时 PCR 分析表明，阈值循环与 P. betae 的数量之间存在密切关系。对播种后 20 天内连续采样的感染植物进行杂交探针实时分析表明，在初始感染后，转录本水平稳步上升至峰值，然后下降。在易感植物和抗性野生贝塔种中的感染比较时程分析表明，尽管易感植物中的转录本水平呈持续上升趋势，但在抗性种中，仅在极低水平下才可检测到。
