Droplet-vitrification cryotherapy and thermotherapy as efficient tools for the eradication of apple chlorotic leaf spot virus and apple stem grooving virus from virus-infected quince in vitro cultures

Farhadi-Tooli, Sakineh; Ghanbari, Atefeh; Kermani, Maryam Jafarkhani; Zeinalabedini, Mehrshad; Bettoni, Jean Carlos; Naji, Amir Mohammad; Kazemi, Nooshin

doi:10.1007/s10658-021-02400-x

Cited by 21 publications

(13 citation statements)

References 62 publications

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“…We found that thermotherapy involving alternating temperature of 28°C (8 h in dark) and 40°C (16 h in light) for 2 weeks followed by the excision of 1 mm shoot tips resulted in very low efficiency of virus eradication in “V500,” which was mixed-infected with PVS and PVM, and failed to eradicate either the single infection of PVS from “Dunluce” or the mixed infection of PVS and PVA from “Tahi.” Although 70% of “Tahi” shoots were free of PVA following this thermotherapy regime, none of the recovered plants were free of PVS. In general, prolonged heat treatment associated with high temperatures can increase the frequency of virus eradication, but at the same time reduces the viability of the treated explants, as host plants are often sensitive to these conditions ( Zhao et al, 2018 ; Wang et al, 2018a ; Farhadi-Tooli et al, 2022 ).…”

Section: Discussionmentioning

confidence: 99%

“…In recent years, shoot tip cryotherapy has proven to be an effective method to eradicate viruses from in vitro infected plants in multiple species of economic importance ( Cui et al, 2015 ; Pathirana et al, 2015 ; Bettoni et al, 2018 , 2019a , 2021a ; Bi et al, 2018 ; Souza et al, 2020 ; Farhadi-Tooli et al, 2022 ; Wang et al, 2022a ). Cryotherapy makes use of LN exposure (−196°C) to selectively destroy vacuolated and differentiated cells that are known to harbor viruses within shoot tips ( Wang et al, 2009 ; Wang and Valkonen, 2009 ).…”

Section: Discussionmentioning

confidence: 99%

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Eradication of Potato Virus S, Potato Virus A, and Potato Virus M From Infected in vitro-Grown Potato Shoots Using in vitro Therapies

et al. 2022

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Certain viruses dramatically affect yield and quality of potatoes and have proved difficult to eradicate with current approaches. Here, we describe a reliable and efficient virus eradication method that is high throughput and more efficacious at producing virus-free potato plants than current reported methods. Thermotherapy, chemotherapy, and cryotherapy treatments were tested alone and in combination for ability to eradicate single and mixed Potato virus S (PVS), Potato virus A (PVA), and Potato virus M (PVM) infections from three potato cultivars. Chemotherapy treatments were undertaken on in vitro shoot segments for four weeks in culture medium supplemented with 100 mg L−1 ribavirin. Thermotherapy on in vitro shoot segments was applied for two weeks at 40°C (day) and 28°C (night) with a 16 h photoperiod. Plant vitrification solution 2 (PVS2) and cryotherapy treatments included a shoot tip preculture followed by exposure to PVS2 either without or with liquid nitrogen (LN, cryotherapy) treatment. The virus status of control and recovered plants following therapies was assessed in post-regeneration culture after 3 months and then retested in plants after they had been growing in a greenhouse for a further 3 months. Microtuber production was investigated using in vitro virus-free and virus-infected segments. We found that thermotherapy and cryotherapy (60 min PVS2 + LN) used alone were not effective in virus eradication, while chemotherapy was better but with variable efficacy (20–100%). The most effective result (70–100% virus eradication) was obtained by combining chemotherapy with cryotherapy, or by consecutive chemotherapy, combined chemotherapy and thermotherapy, then cryotherapy treatments irrespective of cultivar. Regrowth following the two best virus eradication treatments was similar ranging from 8.6 to 29% across the three cultivars. The importance of virus removal on yield was reflected in “Dunluce” free of PVS having higher numbers of microtubers and in “V500’ free of PVS and PVA having a greater proportion of microtubers > 5 mm. Our improved procedure has potential for producing virus-free planting material for the potato industry. It could also underpin the global exchange of virus-free germplasm for conservation and breeding programs.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Eradication of Potato Virus S, Potato Virus A, and Potato Virus M From Infected in vitro-Grown Potato Shoots Using in vitro Therapies

et al. 2022

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“…The efficacy of producing virus-free apple plants varied according to the virus species and the nature of infection (single or mixed), cultivars, and the virus–host combination. Furthermore, the duration and temperature of the thermotherapy, as well as the size of the excised shoot tip affected the extent of virus eradication [ 48 , 51 , 58 , 59 , 60 , 61 ]. For example, Paprštein et al [ 57 ] found that the success of thermotherapy in eradicating ACLSV, ASPV, and ASGV depended on the apple cultivar.…”

Section: Discussionmentioning

confidence: 99%

“…Zhao et al [ 48 ], using the same thermotherapy condition as the present study, found frequencies of ASGV eradication of 20% and 41% in shoot tips (1.5 mm) excised from thermo-treated in vitro shoots of ‘Gala’ for four and six weeks, respectively. In general, prolonged heat treatment associated with high temperatures can increase the frequency of virus eradication, but concurrently reduces the viability of the treated explants, and often the host plants are sensitive to these conditions [ 51 , 57 , 59 , 62 , 63 ].…”

Section: Discussionmentioning

confidence: 99%

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Thermotherapy Followed by Shoot Tip Cryotherapy Eradicates Latent Viruses and Apple Hammerhead Viroid from In Vitro Apple Rootstocks

Bettoni

Fazio²,

Costa

et al. 2022

Plants

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Virus and viroid-free apple rootstocks are necessary for large-scale nursery propagation of apple (Malus domestica) trees. Apple stem grooving virus (ASGV) and Apple chlorotic leaf spot virus (ACLSV) are among the most serious apple viruses that are prevalent in most apple growing regions. In addition to these viruses, a new infectious agent named Apple hammerhead viroid (AHVd) has been identified. We investigated whether thermotherapy or cryotherapy alone or a combination of both could effectively eradicate ACLSV, ASGV, and AHVd from in vitro cultures of four apple rootstocks developed in the Cornell-Geneva apple rootstock breeding program (CG 2034, CG 4213, CG 5257, and CG 6006). For thermotherapy treatments, in vitro plants were treated for four weeks at 36 °C (day) and 32 °C (night). Plant vitrification solution 2 (PVS2) and cryotherapy treatments included a shoot tip preculture in 2 M glycerol + 0.8 M sucrose for one day followed by exposure to PVS2 for 60 or 75 min at 22 °C, either without or with liquid nitrogen (LN, cryotherapy) exposure. Combinations of thermotherapy and PVS2/cryotherapy treatments were also performed. Following treatments, shoot tips were warmed, recovered on growth medium, transferred to the greenhouse, grown, placed in dormancy inducing conditions, and then grown again prior to sampling leaves for the presence of viruses and viroids. Overall, thermotherapy combined with cryotherapy treatment resulted in the highest percentage of virus- and viroid-free plants, suggesting great potential for producing virus- and viroid-free planting materials for the apple industry. Furthermore, it could also be a valuable tool to support the global exchange of apple germplasm.

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Assessment of virus eradication methods from infected in vitro-grown apricot cultures

Zare Khafri,

Zarghami,

Naderpour

et al. 2024

Plant Cell Tiss Organ Cult

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Droplet-vitrification cryotherapy and thermotherapy as efficient tools for the eradication of apple chlorotic leaf spot virus and apple stem grooving virus from virus-infected quince in vitro cultures

Cited by 21 publications

References 62 publications

Eradication of Potato Virus S, Potato Virus A, and Potato Virus M From Infected in vitro-Grown Potato Shoots Using in vitro Therapies

Eradication of Potato Virus S, Potato Virus A, and Potato Virus M From Infected in vitro-Grown Potato Shoots Using in vitro Therapies

Thermotherapy Followed by Shoot Tip Cryotherapy Eradicates Latent Viruses and Apple Hammerhead Viroid from In Vitro Apple Rootstocks

Assessment of virus eradication methods from infected in vitro-grown apricot cultures

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