Abstract
BackgroundPreservation of plant virus is a fundamental requirement in all types of virus-related research and applied applications. Development of efficient, reliable strategies for long-term preservation of plant virus would largely assist these studies.ResultsThe present study reported a novel biotechnology allowing cryopreservation of Apple stem grooving virus (ASGV) in living shoot tips. Following cryopreservation by droplet-vitrification or encapsulation-dehydration, about 62–67% of shoot regrowth and 100% of ASGV cryopreservation were obtained. Although shoot proliferation and virus concentration were reduced in cryopreserved diseased shoots after 8 weeks of shoot regeneration, continuous subculture for 4 times (16 weeks) increased shoot proliferation and virus concentration to comparative levels as those produced by shoot tip culture (as a control to shoot tip cryopreservation). Cryopreserved ASGV was efficiently transmitted to a woody plant by micrografting and to a herbaceous indicator by mechanical inoculation. Gene sequencing in three fragments of ASGV genome including coat protein and movement protein showed that cryopreserved ASGV shared 99.87% nucleotide identities with shoot tip culture-preserved virus, indicating cryopreserved virus is genetically stable.ConclusionsThe present study demonstrates ASGV, a representative virus that can infect meristematic cells of shoot tips, can be efficiently cryopreserved in shoot tips. To the best of our knowledge, this is the first report on plant virus cryopreservation in living tissues, and has great potential applications to long-term preservation of plant viruses.
Highlights
Preservation of plant virus is a fundamental requirement in all types of virus-related research and applied applications
Shoot number and length were similar in Apple stem grooving virus (ASGV)-infected shoots regenerated from droplet-vitrification and encapsulation-dehydration cryopreservation, which were significantly lower in ASGV-infected shoots regenerated from shoot tip culture in the first and second subculture (Table 2)
No differences were found in shoot number and length in ASGV-infected shoots regenerated from droplet-vitrification and encapsulationdehydration cryopreservation, and shoot tip culture after the fourth time of subculture (Table 2, Fig. 1c–e)
Summary
Preservation of plant virus is a fundamental requirement in all types of virus-related research and applied applications such as antigen preparation for virus detection by immunology-based methods [1], production of vaccines [2, 3], genetic transformation to produce. Fan et al [15] reported preservation of viral genomes in 700-y-old caribou feces from a subarctic ice patch Their results indicate virus can be cryopreserved for decades. In the study of De Wijs and Suda-Bachmann [19], particles of Potato virus Y (PVY) and Watermelon mosaic virus (WMMV) were cryopreserved in LN for 22 months for the former and 32 months for the latter, without any decreases in the virus infectivity These data indicate cryostorage of virus seems a very promising long-term preservation method for plant viruses
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