Papura D, Jacquot E, Dedryver C A, Luche S, Riault G, Bossis M, Rabilloud T
INRA/ENSA, UMR BiO3P, Le Rheu, France.
Arch Virol. 2002 Oct;147(10):1881-98. doi: 10.1007/s00705-002-0859-6.
Aset of 39 F1 Sitobion avenae clones was obtained by selfing a poorly efficient BYDV-PAV vector clone. These clones were genetically typed by 11 microsatellite loci, and tested for BYDV-PAV4 transmission to barley. The 39 clones displayed a continuum in transmission percentages, from 0% to 88% with a significant clone effect. From this set, two highly efficient (HEV) and two poorly efficient (PEV) vectoring clones were more precisely characterized for transmission of two other PAV isolates. The molecular bases of the lower transmissibility of BYDV-PAV4 by PEV clones and of the aphid vectoring properties were investigated respectively by comparing the sequences corresponding to structural proteins (CP and RTD) of BYDV, and by using proteomic analysis of aphids in two dimensional electrophoresis (2-DE) with immobilized pH gradients (IPG) after an improved protein extraction. Four residues specific to BYDV-PAV4 located in the CP sequence (A(24) and L(130)) or in the RTD region (M(334) and S(456)) could be responsible for the lower transmissibility of this isolate by PEV clones. Among a total of 2150 well-resoluted spots scored on S. avenae proteinic pattern, only twelve proteins were qualitatively or quantitatively different between clones. Four out of them discriminated HEV and PEV groups.
通过对一个传播效率较低的大麦黄矮病毒PAV株系(BYDV - PAV)载体克隆进行自交,获得了一组39个麦长管蚜F1克隆。利用11个微卫星位点对这些克隆进行基因分型,并检测它们对BYDV - PAV4向大麦传播的能力。这39个克隆的传播率呈现出从0%到88%的连续变化,且克隆效应显著。从这一组克隆中,更精确地鉴定了两个高效(HEV)和两个低效(PEV)载体克隆对另外两个PAV分离株的传播情况。分别通过比较与BYDV结构蛋白(CP和RTD)对应的序列,以及在改进蛋白质提取后,利用固定化pH梯度(IPG)二维电泳(2 - DE)对蚜虫进行蛋白质组分析,研究了PEV克隆对BYDV - PAV4传播能力较低的分子基础以及蚜虫载体特性。位于CP序列(A(24)和L(130))或RTD区域(M(334)和S(456))的四个BYDV - PAV4特定位点可能是导致PEV克隆对该分离株传播能力较低的原因。在对麦长管蚜蛋白质图谱上总共2150个分辨率良好的斑点进行评分时,各克隆之间只有12种蛋白质在定性或定量上存在差异。其中有4种蛋白质能够区分HEV和PEV组。