Analysis Of Paulownia Plants With Witches' Broom Tissue Culture Seedling Treated With Rifampicin Using SSR Technique

Paulownia SSRs were screened by transfering other woody plants SSRs to Paulownia. On this basis, the effects of Rifampicin on the tomentosa×Paulownia fortunei tissue culture seedling with Witches'broom, Paulownia tomentosa seedling with Witches'broom and tissue culture seedling with Witches'broom were studied respectively. The three Paulownia genomic DNAs in the healthy seedlings, seedlings with witches' broom and seedlings with witches' broom treated by different concentrations of rifampicin were separately amplified by 73 primer pairs. The results were shown as follows:1. 73 pairs of paulownia SSR primers of strip clear, good repeatability were screened from 416 pairs of SSR from Populus,Pinus,chestnut,Walnut,malum,Prunus persica,Pyrus, the ratio of primers which could be successfully amplified on the Paulownia was 20.67%. 73 pairs of SSR primers were used to analyze the 12 clones Paulownia. It was found that only primer PMGC684 showed polymorphism, and clone named "9503" were able to be distinguished from other 11 clones. The other 72 pairs of primers did not show polymorphism.2. 73 pairs of SSR primers were used to amplify the healthy and with witches' broom Paulownia genomic DNA of Paulownia tomentosa×Paulownia fortunei, Paulownia tomentosa and Paulownia fortunei respectively, as a result, 73 primer pairs amplified the healthy and with witches' broom Paulownia genomic DNAs of the three Paulownia consistently.3.The three paulownia with witches' broom was treated by different concentrations rifampicin, the results showed that, with the concentration of rifampicin increased, the witches' broom symptoms gradually disappear, and the three paulownia with witches' broom transform toward healthy. however, the time of the first rooting delayed, when the concentration of rifampicin reach to 150mg·L^-1, the seedlings increasingly weak. It indicat that the rifampicin also have poisoning effect to Paulownia at the same time it have a effect to Phytoplasma.4. Using Nest-PCR we detected the phytoplasma contents in treated seedling of Paulownia tomentosa×Paulownia fortunei with witches' broom , Paulownia tomentosa with witches' broom and Paulownia fortunei with witches' broom by different concentration rifampicin. The result showed that with the concentrations of rifampicin increasing, the content of Phytoplasma in three Paulownia decreased. when the concentration of rifampicin reach to 150mg·L^-1, we can not detect the presence of phytoplasma.5. The three Paulownia genomic DNAs in the seedlings with witches' broom and seedlings with witches' broom were separately amplified by 73 primer pairs. The results showed that the amplification results was consistent in different treatments.