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大麦抗BYDV的蚜虫获毒酶联免疫吸附法
刘常宏1, S.Haber2
1.陕西省植物保护研究所;2.Agriculture Canada Winnipeg Research Starion
摘要:
蚜虫获毒酶联免疫吸附法(AA-ELISA)是指用ELISA测定从染病植株上获毒蚜虫以反映品种抗BYDV的方法。经对带Yd2基因大麦品种877-16与感病品种Manley对比测定,按其方法接种3叶期第1叶片,传毒3d后用无毒禾谷类蚜虫从第2叶片上取食获毒1d,采集5头作ELISA测定。AA-ELISA法测定OD值,Manley显著高于877-16,用ELISA直接测定无显著差异。用AA-ELISA法具有不破坏植株,可对同一部位重复测定,且蚜虫体软,易研碎抽提病毒,测定抗感品种差异明显等优点,具有适用性。
关键词:  蚜虫获毒酶联免疫吸附法,大麦,BYDV
DOI:
分类号:S512.3
基金项目:“小麦育种及改良”项目资助
Detection of Barley Resistance to BYDV with AA-ELISA
Abstract:
A new methos of APhid Aequisition Enzyme-linked Immunosorbent Assay (AA一ELISA )was established for the distinguish of barley resistance to BYDV according the following procedures:1.to transmit BYDV by the common inoculation method to the first appearance of seeding at the 3一leaf stage;2.to make the infceted seedings clearly after 3 days transmision;3.to put non一virus aPhids into an artifieial cage and then fixed them on the second leaf for 1 day acquisition;4.to colleet 5 viruliferous aphids per seeding. The OD value of AA一ELISA was significantly different between 877一16 carrying the Yd2 for BYDV resistance and the susceptible Manley.The virus titre of Manley was much higher than that of 877一16.In parallel experiments where leaf tissue was sacrificed for analysis of virus titre by ELISA,877一1 could not be distinguished from Manley beyong 6 days after inoculation.The results indicated that the AA一ELISA was reliable for detecting barley for resistance to BYDV Advantages of AA一ELISA are:it can notdestroy the test tissue and enable the repeat virus titre deteetions on a fixed position of plant;soft aphid tissue is easier to grind and extract than leaf ones;clearer distinctions can obtain between resistant and susceptible varieties.
Key words:  AA-ELISA,barley,BYDV

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