【作者】 任亚萍；

【导师】 陈龙清；

【作者基本信息】 华中农业大学 ， 园林植物与观赏园艺， 2008， 硕士

【摘要】 以麝香百合（Lilium.longiflorum）、亚洲百合杂交系（Lilium.asiatic hybrids）两个品种‘Acuba’、‘Nove Cento’及卷丹（Lilium.lancifolium）的无菌苗为外植体,从培养条件入手,探索优化促进试管鳞茎膨大的培养基,为百合试管鳞茎的工厂化生产提供理论依据和关键技术。结论如下:1.以麝香百合的花丝、花梗等为外植体,诱导形成试管苗。研究发现,培养基MS+6-BA0.5mg/L+NAA0.1mg/L适合花丝诱导愈伤和分化,适合花梗诱导愈伤及分化的培养基为MS+6-BA1.0mg/L+NAA0.5mg/L。MS+6-BA1.0mg/L+NAA0.1mg/L最适合无菌苗的继代,繁殖系数最高,为5.29。2.不同培养条件对试管鳞茎膨大的影响。在一定范围内,提高糖浓度会促进鳞茎的膨大,但浓度过高（120g/L）会影响鳞茎的形成,90g/L对于麝香百合和亚洲百合‘Acuba’都是最适合鳞茎化培养的浓度,鳞茎的平均鲜重分别为107.86mg和127.97mg。生长素NAA对于鳞茎的膨大也有促进作用。在培养基中附加0.5mg/L的NAA最适合麝香百合试管鳞茎的膨大,其鳞茎平均鲜重达122.64mg:而对于‘Nove Cento’,2.0mg/L的NAA适合鳞茎膨大培养,获得的鳞茎平均鲜重为124.22mg。在培养基中附加5mg/L的PP₃₃₃可以使麝香百合获得最大鳞茎鲜重,为99.22mg;10mg/L的PP₃₃₃适合‘Nove Cento’的鳞茎膨大培养,获得的平均鳞重为105.06mg。在正交试验中,‘Acuba’可选用90g/L蔗糖、10mg/L PP₃₃₃和0.025mmol/L SA进行鳞茎化培养,获得鳞茎平均鲜重为161.17mg;‘Nove Cento’可选用90g/L蔗糖、1mg/L PP₃₃₃和0.1mmol/L SA进行鳞茎化培养,获得鳞茎平均鲜重为209.93mg。活性炭具有很强的促进生长和生根效果,同时也具有较强的促进鳞茎形成效果,麝香百合和卷丹都可采用3g/L的AC进行鳞茎化培养,获得的鳞茎平均鲜重都为对照的2～3倍。茉莉酸甲酯（Me-JA）对麝香百合鳞茎的增重有效果,可以使鳞茎平均鲜重达到110.77mg;而对于卷丹,茉莉酸甲酯并没有显著作用。液体浅层静置由于更易于鳞茎对营养物质的吸收,因此促进鳞茎的膨大,麝香百合和卷丹在液体培养中都获得了最大的鳞茎鲜重,分别为161.62mg和249.84mg,超过8mm的大鳞茎分别达到48.53%和54.05%,这种培养方式还可以缩短鳞茎培养时间到60天。以液体培养基为基础,添加蔗糖和活性炭能够很有效地促进试管鳞茎膨大,这种培养基会强烈地促使叶片及根生长,还有试管鳞茎的膨大,比单独使用其中任何一种,效果都要好得多。60g/L蔗糖+3g/L活性炭最有利于麝香百合试管鳞茎膨大,平均鲜重达627.32mg,获得的13mm以上的鳞茎达39.02%;90g/L蔗糖+3g/L活性炭是促进卷丹试管鳞茎膨大的最适浓度,平均鲜重达479.17mg,获得的13m以上的鳞茎达10.25%。3.将固体培养基上获得的试管鳞茎按直径大小分成A:8mm以上、B:5-8mm及C:3-5mm三个等级,经播种栽培后麝香百合成活率分别为95.8%、81.9%、80.6%。亚洲百合‘Nove Cento’成活率分别为85.7%、68.3%、44.4%。萌发苗长势随直径的增大而增强。将液体培养基上获得的卷丹试管鳞茎按直径大小分成D:12mm以上、E:9-12mm和F:7-9mm三个等级,经播种栽培后卷丹成活率都在85%以上。12mm以上鳞茎萌发植株最健壮,60天后鳞茎平均直径仍在7mm以上。更多还原

【Abstract】 The sterile seedlings of Lilium,longiflorum、Lilium.asiatic hybrids ’Acuba’、’Nove Cento’ and Lilium.lancifolium were taken as explants to induce the bulblets in vitro.In order to improve factory production of bulblets from sterile seedlings,the tissue culture condition of the technique of bulblets enlargement was optimized.1.Filament and pedicel of L.longiflorum were taken as explants to induce plantlet. The medium of MS+6-BA0.5mg/L+NAA0.1 mg/L and MS+6-BA1.0mg/L+NAA0.5mg/L were suitable to induce the callus and differentiation for the filament and pedice respectively;The medium of MS+6-BA1.0mg/L+NAA0.1 mg/1L showed the best effect for the buds multiplication.2.Effect of culture condition on the bulblets enlargement Within a certain range, raising the concentration of sucrose could increase the diameter of bulblets.But the excessive high concentration（120g/L） could weaken the effect.The concentration of 90g/L was the optimal concentration for the culture of bulblets in vitro,either L.longiflorum or ’Acuba’,the fresh weight of bulblets came up to 107.86mg and 127.97mg respectively.The medium additioned 5mg/L PP₃₃₃ showed the best fresh weight of L.longiflorum bulblets in vitro,it was 99.22mg;for ’Nove Cento’,the concentration of 10mg/L was suitable.The concentration of 0.5mg/L NAA showed the best effect to increasing the fresh weight of L.longiflorum bulblets,it came up to 122.64mg;the concentration 2.0mg/L NAA were suitable for the enlargement of ’Nove Cento’ bulblets.In the experiment of orthogonal design,the medium with 90g/Lsucrose、10mg/L PP₃₃₃ and 0.025mmol/L SA was suitable for ’Acuba’ bulblets enlargement;the medium with 90g/L sucrose、1mg/L PP₃₃₃ and 0.1mmol/L SA was suitable for ’Nove Cento’.AC had powerful effect on promoting leaf growth and rooting,and had powful effect on promoting bulblet enlargement.3g/L AC was suitable for bulblets culture of L. longiflorurn and L.lancifolium,and the fresh weight of them was 2～3 times to CK.Me-JA had powerful effect on L.longiflorum,it made the fresh weight up to 110.77mg;but there was no notable difference on L.lancifolium.The shallow liquid system was useful for the bulblets to absorbe the nutrition,so it was good for the diameter increasing.There was best fresh weight on the shallow liquid system for L.longiflorum and L.lancifolium,it came up to 161.62mg and 249.84mg respectively,and the percentage of large bulblets（dm＞8mm） was 48.53%and 54.05% respectively.This culture system could also shorten the time of bulblets culture to 60 days. The liquid culture containing sucrose and AC simultaneity had more powerful effect on promoting leaf growth,rooting and bulblet enlargement than the effect of using sucrose or AC respectively.The medium with 60g/L sucrose and 3g/L AC showed the best effect to increasing the fresh weight of L.longiflorum bulblets,it came up to 627.32mg, and the percentage of bulblets（dm＞13mm）was up to 39.02%.The medium with 90g/L sucrose and 3g/L AC was suitable for the culture of L.lancifolium bulblets enlargement, the fresh weight of bulblets came up to 479.17mg,and the percentage of bulblets （dm＞13mm） was 10.25%.3.The bulblets from the solid medium were classified to three grades on the basis of diameter A:＞8mm,B:5-8mm and C:3-5mm.The living rate of L.longiflorum and ’Nove Cento’ were respectively 95.8%、81.9%、80.6%and 85.7%、68.3%、44.4%;After sowing, the living seedlings from bigger diameter became stronger.The bulblets from the liquid medium were classified to three grades on the basis of diameter D:＞12mm、E:9-12mm and F:7-9mm.The living rate of L.lancifolium in different grade were up all to 85%.The bulblets whose diameter up to 12mm grew best,and the diameter was over 7mm after planting for 60days.更多还原