摘要
为早期发现桉树幼苗和土壤带菌情况,本试验利用PCR技术对桉树林地土壤及潜伏期桉树青枯菌的检测进行了研究。结果表明:通过SDS法与简单提取法提取土壤中青枯菌DNA,PCR检测灵敏度均达2.5×104CFU/g,简单提取法与SDS法相比具有快速、成本低廉等优点;CTAB法提取人工接菌的桉树组织中青枯菌,PCR检测灵敏度达3×102CFU/g。利用浸泡法处理1年生桉树茎段,用1×107CFU/mL菌悬液处理后,第4天开始发病,至第8天发病率达100%,利用该数据得到了一种制备桉树青枯病潜伏期材料的方法,通过CTAB法提取潜伏期桉树青枯菌的DNA,利用OLI1和Y2引物进行PCR扩增后,检测出288 bp的条带。因此,PCR可以有效地检测土壤和感病潜伏期组织中桉树青枯病菌。
Detection of Ralstonia solanacearum in soil and eucalypt tissue in latent infection based on polymerase chain reaction（PCR） was carried out for discovering whether eucalypt seedlings or soil were contaminated by R.solanacearum.Results showed that the detection limit could be as low as 2.5×104 CFU/g by PCR based on both SDS and Easy extraction methods of DNA.Compared to SDS method,easy extraction method is less time-consuming and inexpensive.The sensitivity for detecting the R.solanacearum in eucalypt tissue can be as low as 3×102 CFU/g.Four days after being dipped in suspension of R.solanacerum at 1×107 CFU/mL,the eucalypt shoot tips started to show typical symptoms of bacterial wilt,and the incidence of wilted shoot tips reached 100% 8 days after inoculation,presenting a successful way to prepare latent infection tissue of eucalypt with R.solanacearum.A specific band with 288 bp could be detected using primer pairs,OLI1 and Y2,to amplify the template DNA extracted by CTAB method from eucalypt tissue in latent infection with R.solanacerum.A conclusion could be drawn that PCR method is a feasible way for detection of cells of R.solanacearum from soil and eucalypt tissues in latent infection.
出处
《河北农业大学学报》
CAS
CSCD
北大核心
2010年第2期79-83,共5页
Journal of Hebei Agricultural University
基金
国家"十一五"林业科技支撑计划(2006BAD08A1102)
国家自然科学基金(30671681)资助
关键词
桉树
青枯病
分子检测
潜伏期
Eucalyptus urophylla
bacterial wilt
molecular detection
latent period
