Bulletin of Botanical Research ›› 2021, Vol. 41 ›› Issue (4): 514-521.doi: 10.7525/j.issn.1673-5102.2021.04.006
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Ren-Rui WANG, Ling-Li TANG, Shi-Lin SU, Cheng-Cai LIU, Jie LI()
Received:
2020-08-07
Online:
2021-07-20
Published:
2021-03-24
Contact:
Jie LI
E-mail:jay0224@sina.com
About author:
WANG Ren-Rui(1981—),female,lecturer,doctor degree,majoring in cryopreservation and cryotherapy of horticultural plants.
Supported by:
CLC Number:
Ren-Rui WANG, Ling-Li TANG, Shi-Lin SU, Cheng-Cai LIU, Jie LI. Elimination of Cymbidium mosaic virus from in vitro Shoot Tips of Phalaenopsis aphrodite by Droplet Vitrification[J]. Bulletin of Botanical Research, 2021, 41(4): 514-521.
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URL: https://bbr.nefu.edu.cn/EN/10.7525/j.issn.1673-5102.2021.04.006
Fig.1
Droplet-vitrification of shoot tips and micropropagation of P. aphrodite ‘Mantianhong’a.A shoot tip(0.5 cm) of Phalaenopsis aphrodite‘Mantianhong’;b.Droplets of 2.5 μL PVS2,each containing a shoot tip,carried on an aluminum foil;c.A survival shoot tip of cryopreservation after 7 d;d.A dead shoot tip of cryopreservation after 7 d;e.A survival shoot tip of cryopreservation after 21 d;f.Protocrom-like bodies regeneration from survival shoot tips after 8 weeks;g.Proliferation of shoots;h.Roots growth of shoots
Table 1
Effect of sucrose concentration in preculture medium on survival rate and regrowth rate of shoot tips(-LN and +LN)
预培养的蔗糖浓度 Sucrose concentration | 成活率Survival rate(%) | 再生率Regrowth rate(%) | ||
---|---|---|---|---|
对照茎尖 -LN | 超低温茎尖 +LN | 对照茎尖 -LN | 超低温茎尖 +LN | |
BM+0.3 mol·L-1蔗糖 | 93.3±3.3x | 43.3±3.3b | 73.3±3.3x | 10.0±5.8bc |
BM+0.6 mol·L-1蔗糖 | 86.7±6.7x | 76.7±3.3a | 66.7±6.7x | 56.7±3.3a |
BM+0.8 mol·L-1蔗糖 | 83.3±3.3x | 70.0±5.8a | 46.7±3.3y | 30.0±5.8b |
BM+1.0 mol·L-1蔗糖 | 56.7±3.3y | 66.7±3.3a | 33.3±3.3y | 6.7±3.3c |
Table 2
Effect of sucrose duration in preculture medium on survival rate and regrowth rate of shoot tips(-LN and +LN)
预培养时间 Preculture duration(d) | 成活率(%) Survival rate | 再生率(%) Regrowth rate | ||
---|---|---|---|---|
对照茎尖 -LN | 超低温茎尖 +LN | 对照茎尖 -LN | 超低温茎尖 +LN | |
1 | 86.7±6.7x | 76.7±3.3a | 66.7±6.7x | 56.7±3.3a |
2 | 80.0±5.8xy | 70.0±5.8a | 60.0±5.8x | 53.3±3.3a |
3 | 73.3±3.3xy | 66.7±3.3a | 53.3±6.7xy | 30.0±5.8b |
4 | 60.0±0y | 20.0±5.8b | 33.3±3.3y | 6.7±3.3c |
1 | 陈和明,吕复兵,肖文芳,等.蝴蝶兰与火焰兰远缘杂交育种初探[J].中国农业大学学报,2019,24(8):60-71. |
Chen H M,Lv F B,Xiao W F,et al.Preliminary study on the intergeneric hybridization breeding between Phalaenopsis and Renanthera[J].Journal of China Agricultural University,2019,24(8):60-71. | |
2 | Lam S H,Hung H Y,Yang M L,et al.Chemical constituents from Phalaenopsis hybrids and their bioactivities[J].Natural Product Communications,2019,14(5),doi:10.1177/1934578X19850688. |
3 | 杜君,王利民,孟月娥,等.我国蝴蝶兰产业发展现状及对策研究[J].北方园艺,2015(9):172-175. |
Du J,Wang L M,Meng Y E,et al.Study on the development status and countermeasures of Phalaenopsisamabilis industry in China[J].Northern Horticulture,2015(9):172-175. | |
4 | 贺嘉.蝴蝶兰(Phalaenopsis)脱毒快繁关键技术及其生理基础的研究[D].南京:南京农业大学,2010. |
He J.Study on key technology and its physiological basis of virus elimination and rapid propagation of Phalaenopsis[D].Nanjing:Nanjing Agricultural University,2010. | |
5 | 郑荔丹,张宏宇,江赜如,等.蝴蝶兰脱毒技术研究进展与展望[J].分子植物育种,2018,16(2):541-545. |
Zheng L D,Zhang H Y,Jiang Z R,et al.Progress and prospect of detoxification technology research in phalaenopsis[J].Molecular Plant Breeding,2018,16(2):541-545. | |
6 | Stancato G C.Virus-free Phalaenopsis production using infected stock plants[J].Plant Cell Biotechnology and Molecular Biology,2016,17(3-4):104-111. |
7 | Kim S M,Baek E,Ryu K H,et al.Differentially expressed genes of Chenopodium amaranticolor in response tocymbidium mosaicvirus infection[J].Virus Research,2016,223:43-51. |
8 | Wang Q C,Valkonen J P T.Cryotherapy of shoot tips:novel pathogen eradication method[J].Trends in Plant Science,2009,14(3):119-122. |
9 | 丁兰,郭艳,董刚,等.国产蝴蝶兰种苗携带建兰花叶病毒(CymMV)和齿兰环斑病毒(ORSV)的调查及脱毒的初步研究[J].北方园艺,2012(2):137-140. |
Ding L,Guo Y,Dong G,et al.Investigation and preliminary study of elimination of CymMV and ORSV from Phalaenopsis in China[J].Northern Horticulture,2012(2):137-140. | |
10 | Chien K W,Agrawal D C,Tsay H S,et al.Elimination of mixed ‘Odontoglossum ringspot’ and ‘Cymbidium mosaic’viruses from Phalaenopsis hybrid ‘V3’ through shoot-tip culture and protocorm-like body selection[J].Crop Protection,2015,67:1-6. |
11 | Albouy J,Flouzat C,Kusiak C,et al.Eradication of orchid viruses by chemotherapy from in- vitro cultures of Cymbidium[J].Acta Horticulturae,1988,234:413-420. |
12 | Lim S T,Wong S M,Goh C J.Elimination of Cymbidium mosaic virus and Odontoglossum ringspot virus from orchids by meristem culture and thin section culture with chemotherapy[J].Annals of Applied Biology,1993,122(2):289-297. |
13 | 姜玲,张明涛,陈泽雄,等.墨兰组织培养结合化学处理脱除建兰花叶病毒(CymMv)的研究[J].园艺学报,2005,32(6):1056-1060. |
Jiang L,Zhang M T,Chen Z X,et al.Eradication of CymMv from tissue cultures of Cymbidium sinense with chemotherapy[J].Acta Horticulturae Sinica,2015,32(6):1056-1060. | |
14 | 靖晶.蝴蝶兰组织培养与脱除病毒技术研究[D].北京:北京林业大学,2011. |
Jing J.Studies on tissue culture and virus elimination techniques of the Phalaenopsis[D].Beijing:Beijing Forestry University,2011. | |
15 | Chen S Y,Cheng L C,Chen C W,et al.NIR-assisted orchid virus therapy using urchin bimetallic nanomaterials in Phalaenopsis[J].Advances in Natural Sciences:Nanoscience and Nanotechnology,2013,4(4):045006. |
16 | Skiada F G,Maliogka V I,Katis N I,et al.Elimination of Grapevine rupestris stem pitting-associated virus(GRSPaV) from two Vitis vinifera cultivars by in vitro chemotherapy[J].European Journal of Plant Pathology,2013,135(2):407-414. |
17 | Venturieri G A,De Arbieto E A M.Ex-vitro establishment of Phalaenopsisamabilis seedlings in different substrates[J].Acta Scientiarum.Agronomy,2011,33(3):495-501. |
18 | Wang B,Wang R R,Cui Z H,et al.Potential applications of cryogenic technologies to plant genetic improvement and pathogen eradication[J].Biotechnology Advances,2014,32(3):583-595. |
19 | Li B Q,Feng C H,Hu L Y,et al.Shoot tip culture and cryopreservation for eradication of Apple stem pitting virus(ASPV) and Apple stem grooving virus(ASGV) from apple rootstocks ‘M9’ and ‘M26’[J].Annals of Applied Biology,2016,168(1):142-150. |
20 | 王仁睿,李杰.建兰花叶病毒和齿兰环斑病毒的检测技术及脱毒方法研究进展[J].中国农学通报,2020,36(11):56-62. |
Wang R R,Li J.Cymbidium mosaic virus and Odontoglossum ringspot virus:research progress on detection techniques and virus elimination[J].Chinese Agricultural Science Bulletin,2020,36(11):56-62. | |
21 | Wolkers W F,Oldenhof H.Cryopreservation and freeze-drying protocols[M].New York:Springer,2015. |
22 | Sakai A,Kobayashi S,Oiyama I.Cryopreservation of nucellar cells of navel orange(Citrus sinensis Osb.var.brasiliensis Tanaka) by vitrification[J].Plant Cell Reports,1990,9(1):30-33. |
23 | 梁芳,邓祖丽颖,许申平,等.蝴蝶兰2种病毒的同步检测及其CP融合反义表达载体构建[J].西南农业学报,2018,31(8):1622-1627. |
Liang F,Deng Z L Y,Xu S P,et al.Simultaneous detection of two viruses and construction of double-antisense expression vector of CP genes in Phalaenopsis[J].Southwest China Journal of Agricultural Sciences,2018,31(8):1622-1627. | |
24 | 聂菁,刘丽凤,任海虹,等.蝴蝶兰类原球茎诱导、增殖及植株再生条件初步研究[J].山西大学学报:自然科学版,2016,39(2):318-324. |
Nie J,Liu L F,Ren H H,et al.Preliminary study on multiplication and regeneration system of protocorm-like bodies in Phalaenopsis[J].Journal of Shanxi University:Natural Science Edition,2018,31(8):2016,39(2):318-324. | |
25 | 许传俊,黄珺梅,曾碧玉,等.病毒唑对大花蕙兰CyMV及ORSV脱毒效果初探[J].亚热带植物科学,2012,41(1):7-9. |
Xu C J,Huang J M,Zeng B Y,et al.Influence of ribavirin on Cymbidium hybridum virus eradication[J].Subtropical Plant Science,2012,41(1):7-9. | |
26 | 李正民.蝴蝶兰无毒苗组培快繁技术研究[D].海口:海南大学,2013. |
Li Z M.Study on rapid propagation of Phalaenopsis virus-free plantlets in vitro[D].Haikou:Hainan University,2013. | |
27 | 刘福秀,王安石,韩玉春,等.文心兰2种主要病毒的快速检测及其组培脱毒方法[J].热带作物学报,2014,35(1):147-151. |
Liu F X,Wang A S,Han Y C,et al.Rapid detection of 2 main viruses in Oncidium and the method of virus elimination by plant tissue culture[J].Chinese Journal of Tropical Crops,2014,35(1):147-151. | |
28 | Brison M,De Boucaud M T,Pierronnet A,et al.Effect of cryopreservation on the sanitary state of a cv Prunus rootstock experimentally contaminated with plum pox potyvirus[J].Plant Science,1997,123(1-2):189-196. |
29 | Helliot B,Panis B,Poumay Y,et al.Cryopreservation for the elimination of cucumber mosaic and banana streak viruses from banana(Musa spp.)[J].Plant Cell Reports,2002,20(12):1117-1122. |
30 | 曾继吾,牛王翠,黄永红,等.利用超低温保存方法脱除香蕉束顶病毒的研究[J].植物遗传资源学报,2009,10(3):457-460. |
Zeng J W,Niu W C,Huang Y H,et al.Elimination ofbanana bunchy top virus(BBTV) by cryopreservation of in vitro-grown shoot tips from banana(Musa spp.)[J].Journal of Plant Genetic Resources,2009,10(3):457-460. | |
31 | Wang Q C,Mawassi M,Li P,et al.Elimination of grapevine virus A(GVA) by cryopreservation of in vitro-grown shoot tips of Vitis vinifera L.[J].Plant Science,2003,165(2):321-327. |
32 | Bayati S,Shams-Bakhsh M,Moieni A.Elimination of grapevine virus A (GVA) by cryotherapy and electrotherapy[J].Journal of Agricultural Science & Technology,2011,13(3):443-450. |
33 | Wang Q C,Liu Y,Xie Y H,et al.Cryotherapy of potato shoot tips for efficient elimination of potato leafroll virus(PLRV) and potato virus Y(PVY)[J].Potato Research,2006,49(2):119-129. |
34 | Dhital S P,Lim H T,Manandhar H K.Elimination of potato viruses(PLRV and PVY) by cryopreservation of in vitro grown shoot tips of potato(Solamum tuberosum L.)[J].Horticulture Environment and Biotechnogy,2009,50(3):233-239. |
35 | 白建明,陈晓玲,卢新雄,等.超低温保存法去除马铃薯X病毒和马铃薯纺锤块茎类病毒[J].分子植物育种,2010,8(3):605-611. |
Bai J M,Chen X L,Lu X X,et al.Elimination of potato virus Xand potato spindle tuber viroid by cryopreservation[J].Molecular Plant Breeding,2010,8(3):605-611. | |
36 | Romadanova N V,Mishustina S A,Omasheva M Y,et al.Cryotherapy as a method for reducing the virus infection of apples(Malus sp.)[J].Cryobiology,2015,71(3):559. |
37 | Ruiz-Sáenz D R,Ayala-Hernández D D,Niino T,et al.Salicylic acid-cryotherapy treatment for elimination of potato virus S from Solanum tuberosum[J].American Journal of Potato Research,2019,96(3):225-234 |
38 | 王仁睿,王乔春,李明福,等.植物茎尖超低温技术脱除病原体研究进展[J].植物检疫,2012,26(4):65-69. |
Wang R R,Wang Q C,Li M F,et al.Advancement on elimination of plant pathogen by cryotherapy of shoot tips[J].Plant Quarantine,2012,26(4):65-69. | |
39 | 王仁睿.菊花、枣和马铃薯种质资源超低温保存技术与脱毒的研究[D].杨凌:西北农林科技大学,2014. |
Wang R R.Cryopreservation for germplasm conservation and pathogen eradication in chrysanthemum,Chinese jujube and potato[D].Yangling:Northwest A&F University,2014. |
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