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Efficiency of eradication of Raspberry bushy dwarf virus from infected raspberry ( Rubus idaeus ) by in vitro chemotherapy, thermotherapy and cryotherapy and their combinations
Plant Cell, Tissue and Organ Culture ( IF 3 ) Pub Date : 2020-04-20 , DOI: 10.1007/s11240-020-01829-y
Liya Mathew , Heather Tiffin , Zoe Erridge , Andrew McLachlan , Donald Hunter , Ranjith Pathirana

Raspberry bushy dwarf virus (RBDV) is one of the few horizontally transmitted plant viruses and is difficult to eradicate from infected raspberry (Rubus idaeus) by conventional methods because of its ability to infect meristematic cells in the apical dome. This virus is present where ever raspberry is grown and its adverse effects on quality and yield of the crop has necessitated development of methods to eradicate RBDV from infected stock plants. We applied ribavirin treatment (30 mg/L for 2 weeks—chemotherapy), thermotherapy (24 °C for 8 h in dark and 39 °C for 16 h with light for 2 weeks), cryotherapy (droplet vitrification of shoot tips) and their different combinations to assess the efficiency of virus eradication from an infected raspberry genotype. Virus testing was done on plantlets in tissue culture after 3–4 months following plant regeneration and repeated in half the population of virus-free plants in the greenhouse, 1 year after the initial test. Results showed that ribavirin treatment severely affected plant regeneration, resulting in low efficiency of virus eradication (10.6% regenerated plantlets free of RBDV out of all explants treated). When it was combined with thermotherapy, plant regeneration was significantly improved, resulting in higher efficiency of virus eradication (44.4%). When the shoot tips subjected to these two treatments were cryopreserved and regenerated, virus eradication efficiency further improved to 58.5%. The control of this treatment where plant material was not treated in liquid nitrogen produced the highest efficiency of virus eradication (78.8%), partly because of better plant regeneration. Thermotherapy and cryotherapy when applied alone failed to remove the virus from infected material; however, when combined, 48.6% efficiency of virus eradication was recorded. Inclusion of in vitro thermotherapy in combination with chemotherapy or cryotherapy or both is important to improve efficiency of RBDV eradication from infected raspberry. Nevertheless this virus can be eradicated by different approaches, and our results are useful for planning virus eradication programmes based on the efficiency of eradication. It is important to test the virus status after greenhouse acclimation, as virus may be suppressed and not completely eliminated in some treatments at tissue culture stage, as shown in ribavirin treatment in our research.



中文翻译:

通过体外化学疗法,热疗法和冷冻疗法及其组合从被感染的覆盆子(Rubus idaeus)上清除覆盆子矮化病毒的效率

覆盆子矮矮化病毒(RBDV)是为数不多的水平传播植物病毒之一,很难从被感染的覆盆子中根除(Rubus idaeus),因为它能够感染顶圆顶中的分生细胞。这种病毒存在于任何覆盆子生长的地方,并且它对农作物的品质和产量产生不利影响,因此需要开发从受感染的植物中根除RBDV的方法。我们应用了利巴韦林治疗(30 mg / L,持续2周-化学治疗),热疗(在黑暗中24°C持续8 h,在光照下39°C持续2 h),冷冻疗法(茎尖的液滴玻璃化)及其不同的组合来评估从感染的树莓基因型清除病毒的效率。在植物再生后的3-4个月后,对组织培养中的小植株进行病毒测试,并在初始测试后1年对温室中一半的无病毒植物种群进行重复测试。结果表明,利巴韦林处理严重影响了植物的再生,导致病毒清除效率低下(所有处理过的外植体中没有RBDV的再生小植株有10.6%)。当与热疗法相结合时,植物的再生得到了显着改善,从而消除病毒的效率更高(44.4%)。将经过这两种处理的芽尖冷冻保存并再生后,根除病毒的效率进一步提高到58.5%。在不使用液氮处理植物材料的情况下,对这种处理方法的控制产生了最高的病毒清除效率(78.8%),部分原因是更好的植物再生能力。单独使用热疗和冷冻疗法无法从感染的材料中清除病毒;但是,相加时为48。记录了6%的病毒清除效率。将体外热疗与化学疗法或冷冻疗法或两者结合使用,对于提高从感染树莓中清除RBDV的效率非常重要。但是,可以通过不同方法根除该病毒,我们的结果对于基于根除效率计划病毒根除计划很有用。正如我们研究中的利巴韦林处理所示,在温室培养后测试病毒状态非常重要,因为在某些组织培养阶段的处理中,病毒可能会被抑制并且不能完全消除。并且我们的结果对于基于根除效率计划病毒根除计划很有用。正如我们研究中的利巴韦林处理所示,在温室培养后测试病毒状态非常重要,因为在某些组织培养阶段的处理中,病毒可能会被抑制并且不能完全消除。并且我们的结果对于基于根除效率计划病毒根除计划很有用。正如我们研究中的利巴韦林处理所示,在温室培养后测试病毒状态非常重要,因为在某些组织培养阶段的处理中,病毒可能会被抑制并且不能完全消除。

更新日期:2020-04-22
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