【作者】 梁艳；

【导师】 陈典；

【作者基本信息】 东北农业大学 ， 蔬菜学， 2005， 硕士

【摘要】 以大蒜茎尖试管苗为外植体,探索试管微鳞茎培养技术,研究微鳞茎形成和膨大过程中内源激素的变化规律。为大蒜脱毒种苗试管微鳞茎的工厂化生产提供理论依据和关键技术。结论如下: 1.试管鳞茎的微繁技术(1)生长素添加不同浓度的NAA 和IBA 均可诱导形成试管微鳞茎并具有较高的鳞茎形成率,其中NAA 对鳞茎的形成更有利。1/2MS +NAA0.1 mg·L-1+IBA1.5mg·L-1 为诱导鳞茎形成、促进鳞茎膨大的最佳激素组合,鳞茎形成率127.78%,鳞茎指数1.21,平均鳞茎鲜重173.2mg。(2)乙烯利不同浓度的乙烯利处理,鳞茎形成率均达到100%以上,其中添加1 mg·L-1效果最佳。鳞茎指数和平均鳞茎鲜重以添加3 mg·L-1为最好。(3)多效唑在一定范围内,随着多效唑浓度的增加,抑制鳞茎的形成与分化,鳞茎发生数减少,地上部生长受抑制严重,叶片和根系变短增粗。添加6mg·L-1 多效唑为诱导鳞茎形成和促进膨大的最佳浓度,鳞茎形成率109.00%,鳞茎鲜重为307.07,鳞茎指数1.15。(4)水杨酸水杨酸各处理鳞茎形成率均在100%以上,鳞茎指数和平均鳞茎鲜重以添加5×10-5 mg·L-1 为最好,鳞茎形成率、鳞茎鲜重、鳞茎指数分别为160.00%,456.77mg 和1.16。(5)蔗糖蔗糖浓度控制在30-120g·L-1范围时,浓度越高,鳞茎形成效果越好,以120 g·L-1处理鳞茎形成率最高。90g·L-1 处理对鳞茎膨大效果最好,鳞茎形成率为147.78%,鳞茎鲜重368.97mg,鳞茎指数1.21。(6)活性炭此处理形成试管微鳞茎历时较长,从处理到鳞茎完全膨大大致要90d,形成初期鳞茎形状近椭圆形,至鳞茎膨大后期才渐呈圆形。3 mg·L-1处理效果最佳,鳞茎形成率、鳞茎指数、鳞茎鲜重分别为,138.89%,1.12 和391.87mg。2.试管微鳞茎形成与膨大过程中内源激素含量及激素间关系的变化(1)蔗糖处理各处理叶鞘中的内源ABA 含量均表现为先增加后降低趋势,随着蔗糖处理浓度的增加,叶鞘内的ABA 含量变化幅度增大。植株叶鞘内的GA 含量变化整体呈现先下降再略有上升趋势,膨大后期GA 含量又有降低倾向,但变化不明显。IAA 含量也呈先高后低的变化规律。提高蔗糖浓度可以促进鳞茎快速形成并促进其尽快地完成膨大期,植物内源激素IAA 有利于鳞茎的形成。内源激素间平衡的关系为:GA/ABA 的值在处理后35d 内急剧下降,在鳞茎进入快速膨大期时,先略增加后趋于稳定,GA/ABA 的比值较低有利于鳞茎的形成。IAA/ABA的比值全程呈现递减的趋势,随蔗糖浓度的变化,二者的比值变化幅度不大,因此,不同处更多还原

【Abstract】 It regarded test-tube seedings of garlic stem tip as explants,then explored the change of cultural technique of test-tube bulb in garlic.At the same time,it studied the change of endogenous hormones during the bulb formation and development,so that it provided theoretical basis and key techniques for the factory production of test-tube bulb from garlic virus-free seedings.The conclusions were as follows: 1.The micropropagation technique of test-tube bulb (1)Auxin Different concentration treatment of NAA and IBA induced bulb formation in vitro,and both had a high bulb formation rate,and the effect of NAA was superior to IBA.The medium of 1/2MS +NAA0.1 mg·L-1+IBA1.5mg·L-1 was fit for the bulb formation and development.The bulbing rate ,bulb index and the fresh weight came up to 127.78%,1.21,173.2mg respectively. (2)Ethephon The bulbing rates of different treatment of Ethephon were over 100%,1mg·L-1 ethephon treatment was the optimal one among of these treatments. As for bulb index and the fresh weight,the treatment of 3 mg·L-1 Ethephon was the optimal. (3)PP333 Within a certain range,raising concentration could inhibit the formation and differentiation of the bulb,the No.of bulb decreased,the growth of the test-tube seedings was heavily inhibited, the leaves and roots were shortened and widened. The 6mg·L-1 PP333 was the optimal treatment concentration of the inducing bulb formation and development, the bulbing rate ,bulb index and the fresh weight came up to 109.00%,1.15,307.07mg respectively. (4)SA The bulbing rates of different treatment of SA were over 100%, As for bulb index and the mean fresh weight,the treatment of 5×10-5mg·L-1 SA was the optimal, the bulbing rate ,bulb index and the fresh weight came up to 160.00%,1.16,456.77mg respectively. (5)sucrose Within the range of 30~120g·L-1,the higher of treatment concentration,the better of the bulb formation effect, 120 g·L-1 sucrose was the optimal which obtained the highest bulbing rate.As for the bulb development, 90 g·L-1 concentration was the optimal, the bulbing rate ,bulb index and the fresh weight came up to 147.78. %,1.21,368.97mg respectively. (6)AC This treatment spent longer time on bulb formation,the whole process from treatment to development was over 90d,and the bulb became round from ellipse gradually during the process of bulb formation and the development. The 3 mg·L-1 treatment was the optimal,he bulbing rate ,bulb index and the fresh weight came up to 138.89%,1.12, 391.87mg respectively. 2.The change of endogenesis hormones content in the leaf sheat and the change of relations among endogenesis hormones during the bulb formation and development (1)sucrose treatment Within the different treatment,the content of ABA rose,then decreased in the leaf sheat, the higher of treatment concentration,the much of the ABA content changed. The content of GA decreased,then rose in the leaf sheat,then decreased during the development period,but the change was not significant.The change of IAA content also rose,then decreased.Raising sucrose concentration could induce bulb formation and development,therefore,IAA was advantage for the bulb formation.The relations among endogenesis hormones were as follows:The number of GA/ABA decreased rapidly during 35d after the threatment,when came to bulb rapid development period,at first rose,then maintain stable, the relatively low number of GA/ABA was advantage for the bulb formation. The number of IAA/ABA decreased through the whole treatment process,with the change of the sucrose concentration,the number changed slightly.Therefore,the differences among treatment had a close relation with the change of the IAA/ABA,while had nothing to do with the change of GA/ABA. (2)PP333 treatment The content of ABA rose,then decreased in the leaf sheat,the increased degree was bigger during the prophase of the treatment.The change of GA content decreased during the bulb formation prophase,then maitain stable.The change of IAA content rose,then decreased,the degree of change was bigger during the 45d,when came to the bulb development period,the content of IAA maitain stable and remain a low level. The relations among endogenesis hormones were follows The change degree of GA/ABA was bigger than the sucrose treatment,the better of the bulb inducing effect was,the more significant of the number decreased.The number of IAA/ABA decreased rapidly during 35d after the threatment,when came to bulb development,the number maitain stable.Therefore,this treatment inducing bulb formation was through the decrease of IAA/ABA and GA/ABA.更多还原