A tissue culture method for the preservation of Soybean mosaic virus strains
A tissue culture method for the preservation of Soybean mosaic virus strains
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DOI：
10.1023/a:1023915932392
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发表时间：
2003-08-01
影响因子：
3
通讯作者：
Veilleux, RE
中科院分区：
文献类型：
--
作者：
Chen, P;Buss, GR;Veilleux, RE
用11株大豆花叶病毒（SMV）在大豆愈伤组织培养中的活性和寿命进行了研究。稳定)。通过在Msoy和MS琼脂培养基上直接培养SMV感染的大豆植株叶片，建立了大豆愈伤组织和SMV双培养。建立的smv愈伤组织在光照下保持在25度，每隔2个月或必要时将其传代到新鲜的MS培养基上，并定期检测病毒感染性。感染愈伤组织在MS培养基上比在Msoy培养基上生长得更好，保持活性的时间更长。在10-15℃条件下，愈伤组织和SMV均可存活13-15周或更长时间。愈伤组织培养的SMV的传染性与感染植株的SMV相当，并且在连续5次继代培养中保持稳定超过一年。双重培养的愈伤组织被SMV均匀感染，从而保证了随机转移的传染性继代培养。通过多次传代培养可以显著提高体外接种量。SMV培养物的生物完整性得以保持，病毒毒力和致病型未发生变化。该方法对于将SMV菌株保存在高传染性和易于获得的形式并减少污染或丧失生存能力的机会具有价值。
The activity and longevity of Soybean mosaic virus (SMV) in soybean callus culture were investigated with 11 SMV strains which are distinguished by differential reactions on soybean cultivars [Glycine max ( L.) Merr.]. Dual cultures (soybean callus and SMV) were initiated by direct culture of SMV-infected leaves from susceptible soybean plants on Msoy and MS agar medium. Established SMV-callus cultures were maintained at 25degreesC under light, subcultured to fresh MS medium at 2-month intervals or as necessary, and assayed periodically for virus infectivity. The infected calluses on MS medium grew better and stayed active longer than those on Msoy medium. At 10-15degreesC, calluses and SMV were viable and active for 13-15 weeks or longer without subculture. The infectivity of SMV from callus cultures was comparable with that of SMV from infected plants, and remained stable for more than a year through five successive subcultures. Callus tissues of dual cultures were uniformly infected by SMV, thus ensuring infectious subcultures by random transfers. Production of in vitro inoculum can be significantly increased by multiple subcultures. Biological integrity of the SMV cultures was maintained with no change of viral virulence and pathotype. The method is of value for preserving a collection of SMV strains in a highly infectious and readily available form and reducing the chance of contamination or loss in viability.