摘要
为确定在新疆地区紫花苜蓿(Medicago sativa)上发生的疑似丛枝病的病害种类,本研究提取了193个疑似紫花苜蓿丛枝病植株的总DNA,并以报道的植原体检测通用引物R16mF2/R16mR1和R16F2/R16R2为引物对其进行了巢式PCR扩增。其中23个样品获得了1.2kb的特异片段,检出率为16.8%,确定该病害为紫花苜蓿丛枝病。构建的系统发育树结果表明,该植原体为16SrⅤ-B亚组成员,与榆树黄化植原体组(Elm Yellows Group,16SrⅤ)中卫矛白化(Euonymus Whitening)植原体的同源性高达99.1%。本研究首次采用分子生物技术确定了新疆紫花苜蓿丛枝病的病原是植原体,明确了其分类地位,该结果可为该病害的早期诊断、检测提供理论依据。
In order to determine the pathogen species of lucerne witches broom disease in Xinjiang,193 lucerne samples with witches broom disease symptom were detected by nested-PCR with R16mF2/R16mR1 and R16F2/R16R2 universal primers for phytoplasma,and 23 samples had positive results with 1.2kb specific fragment,which had detection rate of 16.8% and confirmed that the disease was witches broom.The phylogenetic tree showed that this phytoplasma strain was one member of 16SrⅤ-B subgroup which had 99.1% homology with euonymus whitening phytoplasma in Elm Yellows Group.To the best of authors knowledge,this was the first study to determine phytoplasma pathogen of lucerne witches broom disease in Xinjiang and identify taxonomic status using molecular biological techniques.These results provide theoretical basis for early diagnosis and detection of lucerne witches broom disease.
出处
《草业科学》
CAS
CSCD
北大核心
2016年第6期1183-1188,共6页
Pratacultural Science
基金
新疆维吾尔自治区重点实验室项目--紫花苜蓿"丛枝"状病害的病原研究(XJDX-2012-01)
公益性行业(农业)科研专项经费项目--草地病害防治技术研究与示范(201303057)
国家级大学生创新创业训练计划项目--新疆紫花苜蓿植原体病害的分子检测(201510758040)
关键词
紫花苜蓿
丛枝病
植原体
巢式PCR
系统进化树
Medicago sativa
witches＇ broom
phytoplasma
nested-PCR
phylogenetic tree
