柑橘黄化斑驳相关病毒（Citrus yellow mottle-associated virus，CiYMaV）属马铃薯X病毒属（Potexvirus）、印度柑橘病毒亚属（Mandarivirus），最早于2018年在巴基斯坦被发现，目前该病毒在巴基斯坦多地柑橘产区有发生。本研究首先借助植物双元表达载体pCass4-RZ构建了3个CiYMaV的全长cDNA克隆（pCiYMaV-FL-1、pCiYMaV-FL-18和pCiYMaV-FL-22），随后通过农杆菌真空浸润接种强德勒柚（Citrus grandis）验证其侵染活性。结果表明，在接种40-60天，3个克隆均引起严重的叶脉黄化、叶片斑驳和植株矮化的症状。直接组织点免疫（direct tissue blot immunoassay，DTBIA）和反转录聚合酶链式反应（reverse transcription polymerase chain reaction，RT-PCR）的检测结果表明3个克隆的侵染率为94.7-100%。尽管3个克隆引发的症状相似，pCiYMaV-FL-22在接种60天时，其在植株内的RNA和蛋白积累水平最高。因此，选择pCiYMaV-FL-22将其接种到7种柑橘和3种草本寄主上，结果表明pCiYMaV-FL-22能成功侵染7种柑橘寄主，并且在5个柑橘品种上引起明显症状。通过透射电镜，在接种pCiYMaV-FL-22的柑橘植株组织中观察到长线型、弯曲的病毒颗粒。综上所述，本研究构建了CiYMaV的全长感染性cDNA克隆，完成了该病毒的柯赫氏法则验证，为进一步研究该病毒侵染的分子机制和开发能应用于柑橘上的病毒载体奠定了基础。

Citrus tristeza virus (CTV) is one of the most important causal agents of citrus diseases and exists as numerous strains.CTV is replicated in phloem cells of plants within the family Rutaceae and is transmitted by a few of aphid species. CTVepidemics have caused death of millions of citrus trees in many regions all over the world, where the sour orange (Citrusaurantium) was used as rootstock. Also the production of grapefruit (C. paradisi) and sweet orange (C. sinensis) hasbeen affected by CTV strains. CTV gives uplift to three prominent syndromes, namely quick-decline (tristeza), stempittingand seedling-yellows. The disease is graft-transmissible in nature but not seed-transmitted. However, the tristezadisease in most citrus groves was a man-made problem created by the desire of horticulturists to introduce cultivars fromother citrus growing areas. The utmost importance of the disease called for review articles in numbers of plant protection,epidemiology books, citriculture and proceedings. This review collects the information with respects to disease history,distribution host range, virus isolates association, identification and detection, transmission and management; especiallyon the current status of CTV prevailing and controlling in Pakistan. It provides valuable information for CTV disease andits controlling approaches.