Cryopreservation of virus: a novel biotechnology for long-term preservation of virus in shoot tips.
Cryopreservation of virus: a novel biotechnology for long-term preservation of virus in shoot tips.
复制标题
病毒冷冻保存：一种在茎尖长期保存病毒的新型生物技术
DOI：
10.1186/s13007-018-0312-9
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发表时间：
2018
期刊：
影响因子：
5.1
通讯作者：
Wang QC
中科院分区：
文献类型：
--
作者：
Wang MR;Yang W;Zhao L;Li JW;Liu K;Yu JW;Wu YF;Wang QC
植物病毒的保存是各类病毒研究和应用的基本要求。开发有效的，可靠的策略，长期保存植物病毒将在很大程度上帮助这些studies.ResultsThe本研究报告了一种新的生物技术，允许冷冻保存苹果茎沟病毒（ASGV）在活茎尖。采用液滴玻璃化法和玻璃化脱水法进行超低温保存，可获得62-67%的再生率和100%的ASGV超低温保存率。尽管芽再生8周后，冻存病芽的芽增殖和病毒浓度有所降低，但连续继代4次（16周）使芽增殖和病毒浓度增加到与茎尖培养（作为茎尖冻存的对照）相当的水平。冻存的ASGV有效地传输到木本植物的微嫁接和草本植物的指示器通过机械接种。对ASGV基因组外壳蛋白和运动蛋白3个片段的序列测定表明，ASGV与茎尖培养保存病毒的核苷酸同源性为99.87%，表明ASGV在茎尖中具有遗传稳定性。据我们所知，这是第一次报道植物病毒在活组织中的超低温保存，并有很大的潜在应用，以长期保存植物病毒。
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.
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影响因子：
3
作者：
Chen, P;Buss, GR;Veilleux, RE
通讯作者：
Veilleux, RE
影响因子：
2.9
作者：
Jeon, Su Min;Naing, Aung Htay;Kim, Chang Kil
通讯作者：
Kim, Chang Kil
影响因子：
2.6
作者：
DESEQUEIRA, OA
通讯作者：
DESEQUEIRA, OA
影响因子：
5.9
作者：
Loh HS;Green BJ;Yusibov V
通讯作者：
Yusibov V
影响因子：
4.5
作者：
MacKenzie, DJ;McLean, MA;Green, M
通讯作者：
Green, M