【作者】 王艳军；

【导师】 向长萍； 李锡香；

【作者基本信息】 华中农业大学 ， 蔬菜学， 2003， 硕士

【摘要】 我国是世界上大蒜栽培的第一大国，经过世世代代劳动人民的精心种植、培育，已经形成许多各具特色的地方类型或品种。保存这些类型和品种对我国大蒜业的可持续发展具有重要的战略意义。大蒜属无性繁殖植物，传统的保存方法需要每年进行种质更新，不仅需要耗费大量的人力、财力，还容易受到病虫害和自然灾害的影响。为了克服传统种质资源保存所面临的问题，我们以山东苍山大蒜为材料，进行了大蒜离体保存技术的初步研究，丰要结果如下： (1) 进行了以微繁为基础的大蒜试管苗保存研究。多芽分化试验表明，大蒜茎尖在含有NAA0.1mg/L+6-BA0.1mg/L的培养基中诱导多芽分化的效果最好。将诱导出的芽分离培养成试管苗，接种在15种不同培养基中，在四种不同的温度下保存。结果显示：试管苗在0±2℃和4±2℃，黑暗保存保存6个月，各种处理无死亡现象：15±2℃，光照强度为1080Lux(100FC)(8小时/天)条件生长的试管苗保存6个月，各种处理都生长非常瘦弱，且苗色偏淡，不适于继续保存；20±2℃，光照强度为6130Lux(560FC)(8小时/天)条件下保存6个月，除加有甘露醇和脱落酸的培养基中保存的试管苗更换新鲜培养基可恢复生长外，其他处理的试管苗已不能成活；培养基中添加甘露醇或ABA对试管苗的生长有抑制作用，两者相比，ABA的抑制作用更强烈。 (2) 对大蒜茎尖玻璃化超低温保存的基本条件进行了研究。大蒜茎尖超低温保存的基本程序为，大蒜茎尖经含高浓度蔗糖的培养基预培养后，室温下用60％PVS2预处理一段时间，换用PVS2再处理一段时间，然后将材料直接投入液氮中保存。通过对预处理时间、预处理培养基、材料大小、冰冻保护剂处理时间等的初步研究，结果表明，在大蒜的超低温保存中，当长度为3.0～3.5mm的大蒜茎尖经含0.7mol/L蔗糖的MS培养基预培养5～7天，室温下60％PVS2预处理60min，再用PVS2在0℃下处理30min，冻存无论是2天还是1个月，成活率均达到最大(100％)。更多还原

【Abstract】 Garlic is planted widely in China and has come into being lots of special local types or cultivars by people. It is strategic for the garlic industry sustainable development to keep those garlic types and cultivars. Garlic is one of the vegetative plant and conserved with the traditional method which needs renewing every year. The traditional conservation not only needs labor and money very much, but also is affected by diseases and insect pests, natural disasters. In order to settle those problems, we initiated the study garlic germplasm conservation in vitro using Cangshan garlic. The major results were as follows:(1) The conservation of garlic plantlets that based on micro-propagation was studied. The results showed that medium with 0.1mg/L NAA and 0.1mg/L 6-BA was best for multi-proliferation of garlic shoot-tips. The plantlets that come from the induced buds cultured on 15 different media and conserved at 4 different temperatures for 6 months. The results showed that, conserved at 0±2℃ and 4±2℃, in darkness for 6 months, the plantlets all survived and could continue to be conserved; conserved at 15±2℃ with a 8h/d photoperiod and 1080 Lux for 6 months, the plantlets were slim . light green and not suitable to conservation: conserved at 20±2℃ with a 8h/d photoperiod and 6130 Lux for 6 months, the plantlets all could’t live after being re-cultured except those plantlets which cultured on the media with mannitol and abscisic acid: conserved on media with mannitol or ABA. the plantlets could be restained grow, but the effect of ABA was stronger than mannitol.(2) The basic condition for garlic shoot-tips cryopreservation by vitrification was researched. The procedure obtained was as follows: the shoot-tips of garlic were pre-cultured on medium with high concentration of sucrose, then pretreated with 60% PVS2 at room temperature and treated with PVS2 at 0℃ . at last plunged into liquid nitrogen. The results based on the preliminary studies on pre-culture media, pre-culture time, material size and treatment time of cryprotectants showed that garlic shoot-tips with 3.0 to 3.5mm length that pre-cultured on MS medium with 0.7mol/L sucrose for 5 to 7 days, pre-treated with 60% PVS2 for 60min and treated with PVS2 for 5 to 60min could get highest survival rate after being cryopreserved in liquid nitrogen for 2 dsys or 30 days.更多还原