DNA polymorphism difference between root system and rhizosphere soil arbuscular mycorrhizal fungi of Prunus mume by AFLP analysis

应用巢式PCr并进行梅根系与根围土壤丛枝菌根真菌(ArbuSCulAr MyCOrrIHIzAl fungI,AMf)dnA扩增片段长度多态性(AMPlIfIEd frAgMEnT lEngTH POlyMOrPHISM,AflP)的差异比较,研究梅共生AMf的作用机理。结果表明,从18个梅品种的30个根围土壤样品中有28个样品获得纯化的dnA片段,占样品数93.3%,样品平均多态性位点数为6.5个,nEI’S基因多样性为0.3559±0.1382,SHAnnOn信息指数为0.5299±0.1676。与梅根系AMf dnA多态性比较,根围土壤的平均多态性位点数明显较多;且根系AMf的dnA多态性位点绝大多数存在于土壤AMf的dnA多态性位点中,表明根系内AMf是由土壤AMf发育而来;根系与根围土壤AMf dnA的聚类均与梅品种群、品种关联性不强,表明AMf对宿主梅品种或品种群没有特异的共生关系。DNA polymorphism of arbuscular mycorrhizal fungi(AMF) in root system and those in rhizosphere soil of Prunus mume was analyzed through DNA amplification by nested PCR based on AFLP marker.The results show that the purified DNA can be extracted from 28 soil samples, accounting for 93.3% of totally 30 samples, collected from rhizosphere of 18 cultivars, averaging 6.5 loci for each sample.The average genetic identity was 0.3559±0.1382, Shannon information index was 0.5299±0.1676.The number of loci in rhizosphere soil was much more than that of loci in root sample, and most of loci in root sample could be found in soil sample.It was proved that the AMF in root might be developed from soil AMF.The clustered groups of AMF DNA by AFLP marker both from roots and from soils were not connected with cultivar groups or cultivars of P.mume, and indicated that there was no specific symbiotic relationship between AMF and P.mume.国家自然科学基金(30470006); 厦门市科技项目(3502Z20072010;3502Z20112004)~

AFLP analysis of arbuscular mycorrhizal fungi in roots of Prunus mume

第一作者: 蔡邦平，博士，副研究员。主要研究方向: 园林植物与观赏园艺、植物菌根。电话: 0592--2039576 Email : cbangping@ 163.com 地址: 361003 福建省厦门市思明区虎园路25 号厦门市园林植物园。[中文文摘]为了解决梅根系共生的丛枝菌根(AM)真菌难以应用形态学鉴定的问题,以巢式PCR的AFLP方法研究梅根系AM真菌DNA多态性。试验采集梅花期的根系样品,应用改良CTAB法提取总DNA,经纯化处理后,应用巢式PCR扩增根系AM真菌基因片段,进行AFLP分析。结果表明,18个梅品种的30个根系样品中,仅有8个样品经巢式PCR后获得纯化的DNA片段,占试验样品数的26.7%;8个样品共得到指纹图谱带24条,各样品平均多态性位点数为3.0个,Nei’s基因多样性为0.4097±0.0848,Shannon信息指数为0.5968±0.0955;利用Nei’s遗传相似性系数聚类,梅品种根系内AMF基因组DNA的聚类类别与梅"品种群"这一分类级别无相关性。该试验为植物根系共生AM真菌DNA多态性研究提供了一种简便的技术。[英文文摘]DNA polymorphism of arbuscular mycorrhizal fungi (AMF) was analyzed through the method of DNA amplification by nested PCR based on AFLP marker,in order to solve the difficulty of identifying the species of AMF associated with mei flowers (Prunus mume Sieb.et Zucc.).A total of 30 root samples from 18 mei cultivars were collected in the flowering phase from Wuhan Mei Garden as experimental materials.The results show that the purified DNA can be extracted only from eight root samples that account for 26.7% of total root samples.Totally 24 polymorphic loci were obtained from eight sample roots，averaging 3. 0 loci for each sample.The average genetic identity was 0. 409 7 ±0. 084 8，and the Shannon information index was 0. 596 8 ± 0. 095 5． The clustered groups of AMF DNA by AFLP marker from different cultivars were not identical with cultivar groups of P.mume.The results indicate that the AFLP marker technology is a brief and effective method to study the DNA polymorphism for the AMF in the roots of a plant．教育部科学技术研究重点项目(104034);; 国家自然科学基金项目(30670047);; 厦门市科技计划项目(3502Z20072010、3502Z20112004