The Pineapple mealybug wilt-associated virus (PMWaV) provokes a disease that causes considerable losses to pineapple growers due to the reduced leaf tissue turgescence, resulting in leaf necrosis and death in severe cases. In this work, we describe the comparison of shoot tip culture and cryotherapy methods to eradicate the pineapple wilt disease-associated ampeloviruses. Plants from the accessions Ananas comosus var. comosus (AGB-009), var. bracteatus (AGB-119), and var. parguazensis (AGB-376) were indexed by RT-PCR, confirming mixed infections of PMWaV-1, PMWaV-2, and PMWaV-3. The accessions were cultured in vitro and their shoot tips treated by cryotherapy following a droplet vitrification protocol. The regeneration rate from shoot tip culture was 93% for AGB-376 and 100% for the other two accessions. After freezing, AGB-376 had 100% regeneration subsequent to exposure to PVS2 for 45 min, followed by 95% for AGB-009, while for AGB-119 the optimal exposure time was 60 min, with plant regeneration from nearly 80% of the shoot tips. For the accessions AGB-009 and AGB-376, all the recovered plants were virus free by the two methods, while 50% of the plants from accession AGB-119 remained infected. These results indicate that shoot tip culture alone or in association with cryotherapy is a promising routine method for virus removal from pineapple plant tissues and is useful to ensure that backup reserves of pineapple germplasm, conserved by in vitro bank and cryobank, are formed with virus-free plants.

的菠萝粉蚧萎病毒相关病毒（PMWaV）引发，导致由于减少的叶组织turgescence相当大的损失，以菠萝种植者，从而导致严重的情况下叶坏死和死亡的疾病。在这项工作中，我们描述了根尖培养和冷冻疗法根除菠萝枯萎病相关的两性病毒的比较。凤梨Ananas comosus var。的植物。comosus（AGB-009），变型 bracteatus（AGB-119）和var。通过RT-PCR对帕瓜（AGB-376）进行索引，确认了PMWaV-1，PMWaV-2和PMWaV-3的混合感染。将种质进行体外培养并按照液滴玻璃化方案对它们的梢进行冷冻治疗。对于AGB-376，茎尖培养的再生率为93％，对于其他两种登录种，再生率为100％。冷冻后，暴露于PVS2 45分钟后，AGB-376的再生率为100％，AGB-009暴露为95％，而AGB-119的最佳暴露时间为60分钟，其中近80％的植株再生。拍摄技巧。对于登录号AGB-009和AGB-376，所有回收的植物都通过两种方法不含病毒，而登录号AGB-119的植物中有50％仍被感染。这些结果表明，单独或与冷冻疗法一起进行茎尖培养是从菠萝植物组织中去除病毒的一种有前途的常规方法，并且对于确保菠萝种质的备用储备（可用于保存菠萝种）是有用的。体外库和冷冻库是由无病毒的植物形成的。