Cryopreservation techniques and their application in vegetatively propagated crop plants in Finland

Nukari, Anna; Uosukainen, Marjatta; Rokka, V-M.

doi:10.2137/145960609789267506

Cited by 12 publications

(10 citation statements)

References 44 publications

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“…The results indicate that coinfection with the aphid‐transmitted BRNV and the pollen‐transmitted RBDV is common in wild raspberries in Finland. These findings are important because BRNV has not been reported previously in Finland and because RBDV has been detected so far only in cultivated raspberries in Finland (Wang et al ., ; Nukari et al ., ). Small interfering RNA deep sequencing was used recently to facilitate the first detection of the mite‐transmitted raspberry leaf blotch virus (genus Emaravirus ) in cultivated and wild raspberries in Finland (Dong et al ., ).…”

Section: Discussionmentioning

confidence: 97%

“…RBDV has a wide host range and is known to infect more than 55 plant species in 12 dicotyledonous families (AAB Descriptions of Plant Viruses; http://www.dpvweb.net/dpv/). In Finland, RBDV has been reported to infect cultivated raspberries (Wang et al ., ; Nukari et al ., ) and Arctic brambles, Rubus arcticus L. (Kokko et al ., a , b ), but there are few studies available on occurrence of RBDV in wild raspberries. Finally, raspberry leaf blotch virus (genus Emaravirus ) was also recently detected in Finland in both wild and cultivated raspberries as well as the ornamental species Rubus odoratus L. (Bi et al ., ; Dong et al ., ).…”

Section: Introductionmentioning

confidence: 99%

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Molecular detection and characterisation of black raspberry necrosis virus and raspberry bushy dwarf virus isolates in wild raspberries

Susi

Rajamäki

Artola

et al. 2018

Annals of Applied Biology

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Virus-derived small interfering RNAs (siRNAs) were extracted from leaves of wild raspberries (Rubus idaeus) sampled from three different regions in Finland and subjected to deep sequencing. Assembly of the siRNA reads to contigs and their comparison to sequences in databases revealed the presence of the bipartite positive-sense single-stranded RNA viruses, raspberry bushy dwarf virus (RBDV, genus Idaeovirus), and black raspberry necrosis virus (BRNV, family Secoviridae) in 19 and 26 samples, respectively, including 15 plants coinfected with both viruses. Coverage with siRNA reads [21 and 22 nucleotides (nt)] was higher in BRNV-FI (Finland) RNA1 (79%) than RNA2 (45%). In RBDV, the coverage of siRNA reads was 89% and 90% for RNA1 and RNA2, respectively. Average depth of coverage was 1.6-4.9 for BRNV and 16.5-36.5 for RBDV. PCR primers designed for RBDV and BRNV based on the contigs were used for screening wild raspberry and a few cultivated raspberry samples from different regions. Furthermore, the sequences of BRNV RNA1 and RNA2 were determined by amplification and sequencing of overlapping contigs (length 1000-1200 nt) except for the 3 ′ and 5 ′ ends of RNA1 and RNA2 covered by primers. RNA1 of the Finnish BRNV isolate (BRNV-FI) was 80% and 86% identical to BRNV-NA (USA) and BRNV-Alyth (UK), respectively, whereas the identity of NA and Alyth was 79%. RNA2 of BRNV-FI was 84% and 80% identical to BRNV-NA and BRNV-Alyth, respectively, whereas NA and Alyth were 82% identical. Hence, the strains detected in Finland differ from those reported in the UK and USA. Our results reveal the presence of BRNV in Finland for the first time. The virus is common in wild raspberries and nearly identical isolates are found in cultivated raspberries as well. The results show that wild raspberries in Finland are commonly infected with RBDV or BRNV or both viruses and thus are likely to serve as reservoirs of RBDV and BRNV for cultivated Rubus spp.Rosaceae), which is native to Europe and northern Asia. Hybrids of R. idaeus and the closely related wild North American form known as Rubus idaeus var. strigosus or Rubus strigosus are cultivated as well (Dale et al., 1993).The perennial nature of raspberries and the fact that they propagate vegetatively and produce berries only by the shoots of the second year make the crop prone to Ann Appl Biol 173 (2018) 97-111 97

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Section: Discussionmentioning

confidence: 97%

Section: Introductionmentioning

confidence: 99%

Molecular detection and characterisation of black raspberry necrosis virus and raspberry bushy dwarf virus isolates in wild raspberries

Susi

Rajamäki

Artola

et al. 2018

Annals of Applied Biology

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“…In view of endangered species, where seed production may be low, or species are cross pollinated, this method represents a quick possibility to conserve the genetic diversity of the remnant population. Established in vitro cultures are often preferred as starting material for cryopreservation over field material because of the fear of introducing contamination by microbes (Nukari et al 2009). However, this potential disadvantage of field material never materialized: clean cultures could be established using the described sterilisation method in combination with very small explants that were protected by leaf bases with 100% success.…”

Section: Discussionmentioning

confidence: 99%

Direct cryopreservation of winter-acclimated buds of Dracocephalum austriacum (Lamiaceae) from field material

Rasl

Schalk

Temsch

et al. 2020

Plant Cell Tiss Organ Cult

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This study develops protocols for the micropropagation and cryopreservation of Dracocephalum austriacum (Lamiaceae). It is a perennial herbaceous plant that overwinters with ground-level sprouts and is classified as critically endangered in Europe. In vitro cultures were initiated from seeds on growth-regulator-free Murashige & Skoog (MS) medium after nicking the seed coat. Propagation via shoot culture was achieved on ½ MS medium with 1 µM benzyl adenine (BAP). Rooting on various indole-3-butyric acid (IBA)-media was not reliable, but the rooting success was 80% after 10 weeks on medium with 1 µM BAP. Two starting materials underwent cryopreservation: (1) shoot tips from cold-acclimated in vitro plantlets and (2) axillary buds from winter shoots from field plants. For the cryopreservation of in vitro shoots, plant vitrification solution (PVS)3 and incubation over ice yielded the best results (~ 34% regeneration success). However, regeneration using winter acclimated buds were 100, 76 and 30% for collections in December, February and March, respectively, using the same protocol. Moreover, the ploidy levels of cryopreserved plantlets were estimated using flow cytometry. The use of winteracclimated field material of temperate herbaceous plants or subshrubs has high potential as explant source for cryopreservation and calls for exploring this technique for other species.

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“…Of 40 analyzed plant samples, 17 were found to contain two or three viruses. RBDV and BRNV have been previously identified in both cultivated and wild raspberries in Finland [7, 21, 39, 40]. In addition, RBDV has been detected in arctic brambles ( Rubus arcticus L.) in Finland [39, 41].…”

Section: Discussionmentioning

confidence: 99%

Small-RNA analysis of pre-basic mother plants and conserved accessions of plant genetic resources for the presence of viruses

et al. 2019

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Pathogen-free stocks of vegetatively propagated plants are crucial in certified plant production. They require regular monitoring of the plant germplasm for pathogens, especially of the stocks maintained in the field. Here we tested pre-basic mother plants of Fragaria , Rubus and Ribes spp., and conserved accessions of the plant genetic resources of Rubus spp. maintained at research stations in Finland, for the presence of viruses using small interfering RNA (siRNA) -based diagnostics (VirusDetect). The advance of the method is that unrelated viruses can be detected simultaneously without resumptions of the viruses present. While no virus was detected in pre-basic mother plants of Fragaria and Ribes species, rubus yellow net virus (RYNV) was detected in pre-basic mother plants of Rubus . Raspberry bushy dwarf virus (RBDV), black raspberry necrosis virus (BRNV), raspberry vein chlorosis virus (RVCV) and RYNV were detected in the Rubus genetic resource collection. The L polymerase encoding sequence characterized from seven RVCV isolates showed considerable genetic variation. The data provide the first molecular biological evidence for the presence of RYNV in Finland. RYNV was not revealed in virus indexing by indicator plants, which suggests that it may be endogenously present in some raspberry cultivars. In addition, a putative new RYNV-like badnavirus was detected in Rubus spp. Blackcurrant reversion virus (BRV) and gooseberry vein banding associated virus (GVBaV) were detected in symptomatic Ribes plants grown in the field. Results were consistent with those obtained using PCR or reverse transcription PCR and suggest that the current virus indexing methods of pre-basic mother plants work as expected. Furthermore, many new viruses were identified in the collections of plant genetic resources not previously tested for viruses. In the future, siRNA-based diagnostics could be a useful supplement for the currently used virus detection methods in certified plant production and thus rationalize and simplify the current testing system.

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Cryopreservation techniques and their application in vegetatively propagated crop plants in Finland

Cited by 12 publications

References 44 publications

Molecular detection and characterisation of black raspberry necrosis virus and raspberry bushy dwarf virus isolates in wild raspberries

Molecular detection and characterisation of black raspberry necrosis virus and raspberry bushy dwarf virus isolates in wild raspberries

Direct cryopreservation of winter-acclimated buds of Dracocephalum austriacum (Lamiaceae) from field material

Small-RNA analysis of pre-basic mother plants and conserved accessions of plant genetic resources for the presence of viruses

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