Obura Evans, Midega Charles A O, Masiga Daniel, Pickett John A, Hassan Mohamed, Koji Shinsaku, Khan Zeyaur R
International Centre of Insect Physiology and Ecology, Nairobi, Kenya.
Naturwissenschaften. 2009 Oct;96(10):1169-76. doi: 10.1007/s00114-009-0578-x. Epub 2009 Jul 4.
Napier grass (Pennisetum purpureum) is the most important fodder crop in smallholder dairy production systems in East Africa, characterized by small zero-grazing units. It is also an important trap crop used in the management of cereal stemborers in maize in the region. However, production of Napier grass in the region is severely constrained by Napier stunt disease. The etiology of the disease is known to be a phytoplasma, 16SrXI strain. However, the putative insect vector was yet unknown. We sampled and identified five leafhopper and three planthopper species associated with Napier grass and used them as candidates in pathogen transmission experiments. Polymerase chain reaction (PCR), based on the highly conserved 16S gene, primed by P1/P6-R16F2n/R16R2 nested primer sets was used to diagnose phytoplasma on test plants and insects, before and after transmission experiments. Healthy plants were exposed for 60 days to insects that had fed on diseased plants and acquired phytoplasma. The plants were then incubated for another 30 days. Nested PCR analyses showed that 58.3% of plants exposed to Recilia banda Kramer (Hemiptera: Cicadellidae) were positive for phytoplasma and developed characteristic stunt disease symptoms while 60% of R. banda insect samples were similarly phytoplasma positive. We compared the nucleotide sequences of the phytoplasma isolated from R. banda, Napier grass on which these insects were fed, and Napier grass infected by R. banda, and found them to be virtually identical. The results confirm that R. banda transmits Napier stunt phytoplasma in western Kenya, and may be the key vector of Napier stunt disease in this region.
象草(狼尾草)是东非小农户奶牛生产系统中最重要的饲料作物,其特点是零放牧单元规模较小。它也是该地区用于防治玉米螟虫的重要诱集作物。然而,该地区象草的生产受到象草矮化病的严重制约。已知该病的病原是一种植原体,16SrXI菌株。然而,假定的昆虫传播媒介尚不清楚。我们采集并鉴定了与象草相关的5种叶蝉和3种飞虱,并将它们作为病原体传播实验的候选对象。在传播实验前后,基于高度保守的16S基因,使用由P1/P6-R16F2n/R16R2巢式引物对引发的聚合酶链反应(PCR)来诊断受试植物和昆虫上的植原体。将健康植物暴露于取食过患病植物并感染了植原体的昆虫60天。然后将这些植物再培养30天。巢式PCR分析表明,暴露于宽带叶蝉(半翅目:叶蝉科)的植物中有58.3%的植原体呈阳性,并出现了典型的矮化病症状,而60%的宽带叶蝉昆虫样本同样呈植原体阳性。我们比较了从宽带叶蝉、这些昆虫取食的象草以及被宽带叶蝉感染的象草中分离出的植原体的核苷酸序列,发现它们几乎相同。结果证实,宽带叶蝉在肯尼亚西部传播象草矮化植原体,可能是该地区象草矮化病的关键传播媒介。