Pitet M, Camprubí A, Calvet C, Estaún V
Plant Protection IRTA, Ctra. de Cabrils Km 2, E-08348 Cabrils, Barcelona, Spain.
Mycorrhiza. 2009 Feb;19(2):125-131. doi: 10.1007/s00572-008-0206-1. Epub 2008 Oct 29.
The effects of the different steps of the root staining on the arbuscular mycorrhizal (AM) fungal rDNA extraction and amplification have been assessed. The results obtained using molecular techniques are compared with those obtained from fresh, non-stained leek roots. A modified staining procedure that eliminates heating, the use of hydrochloric acid and trypan blue, has been proved to be the most adequate to observe the AM colonisation in different plant species with/without lignified roots allowing at the same time the subsequent rDNA extraction and amplification from the stained roots. The staining technique decreased the sensitivity of the process and a higher number of roots had to be used to obtain enough material for a positive amplification. The extraction and amplification process was reliable up to 3 days after staining. A week after staining, the amplification was not dependable and after 2 weeks there was no amplification from stained material.
已评估了根部染色不同步骤对丛枝菌根（AM）真菌rDNA提取和扩增的影响。将使用分子技术获得的结果与从新鲜、未染色的韭菜根获得的结果进行了比较。一种改进的染色程序，该程序省去了加热、盐酸和台盼蓝的使用，已被证明最适合观察不同植物物种（无论有无木质化根）中的AM定殖，同时允许随后从染色的根中提取和扩增rDNA。染色技术降低了该过程的灵敏度，必须使用更多数量的根才能获得足够的材料进行阳性扩增。染色后3天内，提取和扩增过程是可靠的。染色一周后，扩增不可靠，两周后染色材料未出现扩增。
