State Key Laboratory of Crop Stress Biology for Arid Areas, College of Horticulture, Northwest A&F University, Yangling, 712100, Shaanxi, China.
State Key Laboratory of Crop Stress Biology for Arid Areas, College of Plant Protection, Northwest A&F University, Yangling, 712100, Shaanxi, China.
Appl Microbiol Biotechnol. 2018 Dec;102(24):10743-10754. doi: 10.1007/s00253-018-9405-7. Epub 2018 Oct 6.
Availability of and easy access to diverse plant viruses and viroids is a prerequisite in applied and basic studies related to viruses and viroids. Long-term preservation of viruses and viroids is difficult. A protocol was described for long-term preservation of potato leafroll virus (PLRV), potato virus S (PVS), and potato spindle tuber viroid (PSTVd) in cryopreserved shoot tips of potato cv. Zihuabai. Shoot regrowth levels following cryopreservation were higher in 1.5 mm-shoot tips (58-60%) than in 0.5-mm-ones (30-38%). All shoots recovered from 0.5-mm-shoot tips were PVS- and PSTVd-preserved, but none of them were PLRV-preserved. Cryopreservation of 1.5-mm-shoot tips resulted in 35% and 100% of PLRV- and PVS- and PSTVd-preserved shoots. Studies on cell survival patterns and virus localization provided explanations to the varying PLRV-preservation frequencies produced by cryopreservation of the two sizes of shoot tips. Although micropropagation efficiencies were low after 12 weeks of subculture following cryopreservation, similar efficiencies were obtained after 16 weeks of subculture in pathogen-preserved shoots recovered from cryopreservation, compared with the diseased in vitro stock shoots (the control). Pathogen concentrations in the three pathogens-preserved shoots analyzed by qRT-PCR were similar to those in micropropagated shoots. The three pathogens cryopreserved in shoot tips were readily transmitted by grafting and mechanical inoculation to potato plants. PLRV, PVS, and PSTVd represent a diverse range of plant viruses and viroid in terms of taxonomy and infectious ability. Therefore, shoot tip cryopreservation opens a new avenue for long-term preservation of the virus and viroid.
不同植物病毒和类病毒的可用性和易于获取是与病毒和类病毒相关的应用和基础研究的前提。病毒和类病毒的长期保存较为困难。本文描述了一种在Cryopreserved 马铃薯 cv. Zihuabai 茎尖中保存马铃薯卷叶病毒（PLRV）、马铃薯 Y 病毒（PVS）和马铃薯纺锤块茎类病毒（PSTVd）的长期保存方法。Cryopreservation 后，1.5mm 茎尖（58-60%）的茎尖再生水平高于 0.5mm 茎尖（30-38%）。从 0.5mm 茎尖恢复的所有茎均保留了 PVS 和 PSTVd，但均未保留 PLRV。Cryopreservation 1.5mm 茎尖可使 35%和 100%的 PLRV、PVS 和 PSTVd 保留的茎尖再生。细胞存活模式和病毒定位研究为两种大小的茎尖 Cryopreservation 产生的不同 PLRV 保存频率提供了解释。尽管 Cryopreservation 后继代培养 12 周后的微繁殖效率较低，但从 Cryopreservation 中恢复的保存有病原菌的茎尖继代培养 16 周后，与患病的体外库存茎尖（对照）相比，获得了相似的效率。qRT-PCR 分析的三种病原菌保存茎尖中的病原菌浓度与微繁殖茎尖中的浓度相似。通过嫁接和机械接种，Cryopreserved 在茎尖中的三种病原菌很容易传播到马铃薯植株中。PLRV、PVS 和 PSTVd 在分类学和感染能力方面代表了广泛的植物病毒和类病毒。因此，茎尖 Cryopreservation 为病毒和类病毒的长期保存开辟了新途径。
