【作者】 常莉；

【导师】 喻方圆； 范国强；

【作者基本信息】 南京林业大学 ， 森林培育， 2009， 硕士

【摘要】 本文首先利用转移扩增法筛选出了适用于泡桐的SSR引物,建立了泡桐的SSR反应体系,在此基础上研究了利福平对丛枝病毛泡桐、丛枝病白花泡桐和丛枝病豫杂一号泡桐的形态及植原体含量的影响,并分别对三种泡桐的健康组培苗、患丛枝病组培苗及用不同浓度利福平处理的患丛枝病组培苗进行了SSR分析,研究结果如下:1.从其它木本植物上的416对SSR引物中筛选出了73对在泡桐上有扩增产物且扩增产物条带清晰、重复性好的引物,在泡桐上成功扩增的引物占所用引物的比率为20.67%。对12个泡桐无性系进行SSR分析,结果表明只有引物PMGC684表现出多态性,能够将泡桐9503与其他11个无性系、品种区分开,其它72对引物未表现出多态性。2.利用筛选出的73对SSR引物分别对患丛枝病毛泡桐和健康毛泡桐、患丛枝病豫杂一号泡桐和健康豫杂一号泡桐、患丛枝病白花泡桐和健康白花泡桐的基因组DNA进行PCR扩增。结果表明,73对SSR引物对三种健康泡桐和患丛枝病泡桐的扩增结果一致。3.用不同浓度的利福平处理三种患丛枝病泡桐的组培苗,结果表明,随着利福平浓度的增加,三种泡桐组培苗的丛枝症状逐渐消失,朝健康状态转化,但生根时间逐渐推迟,当利福平浓度达到150mg·L^-1后,继续加大浓度时,苗木长势逐渐变差,表明利福平在作用于植原体的同时也对寄主泡桐产生了一定的毒害作用。4.用巢式PCR对不同浓度利福平处理的患丛枝病毛泡桐、患丛枝病白花泡桐和患丛枝病豫杂一号泡桐组培苗体内的植原体含量进行检测,结果表明随着利福平浓度的增加泡桐体内的植原体含量逐渐减少,当利福平浓度达到150mg·L^-1时,检测不到植原体的存在。5.利用筛选出的73对SSR引物分别对不同浓度利福平处理的患丛枝病毛泡桐、患丛枝病豫杂一号泡桐和患丛枝病白花泡桐泡桐的基因组DNA进行PCR扩增,结果表明各处理在73个位点上的扩增结果一致。更多还原

【Abstract】 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.更多还原