【作者】 孙安妮；

【导师】 张延龙；

【作者基本信息】 西北农林科技大学 ， 园林植物与观赏园艺， 2011， 硕士

【摘要】 秦巴山区及其毗邻地区在我国来说是野生百合资源分布最丰富的地区之一,该地区总共有11种野生百合资源,这其中包括有野生百合资源的变种在内(郭美等2010)。但是最近几年以来,该地区的野生百合资源的种群的生存和繁衍不断的受到来自于一些商用的购于国外的种球的侵入以及一些外来的植物种群的渗入等方面的威胁,而且这些地区的野生百合资源赖以生存的生态环境也经受到了来自于自然的以及人为的等众多因素的破环,因此,对野生百合种质资源进行保护和保存的工作就显得非常的关键(薛晓娜等2007)。相对于百合的其它的组织培养途径来说,体细胞胚的发生诱导途径是一种具有后代数量多、繁殖速度快、结构较完整、再生率较高等众多的优点的途径(崔凯荣2010)。本实验研究了秦巴山区的野生宜昌百合(Lilium leucanthum)的组织培养体系和体细胞胚胎发生诱导体系,对宜昌百合的体细胞胚胎的发生诱导的各种影响因素做了探究。实验结果如下:1.野生宜昌百合的不定芽的发生诱导的研究:野生宜昌百合的鳞片最佳消毒时间:酒精30s和升汞10min的组合,在此消毒灭菌过程中的污染率是33.33%,死亡率为10%。诱导野生宜昌百合不定芽的最佳激素组合是:MS+1.0mg/L 6-BA+0.3mg/L NAA,其不定芽诱导率是86.67%。不同部位的鳞片诱导分化不定芽能力由高到低的排列顺序为:中层>外层>内层。2.野生宜昌百合的体细胞胚胎的发生诱导的研究:三个不同生态型的宜昌百合体细胞胚胎发生的最佳诱导激素浓度组合有所不同。采自于陕西省镇巴县渔渡乡的宜昌百合体细胞胚胎发生的诱导最佳激素浓度组合是:MS+0.5 mg/L TDZ+1.0 mg/L NAA,其诱导体细胞胚胎发生率为91.43%;采自于陕西省留坝县留侯镇的宜昌百合体细胞胚胎发生的诱导最佳激素浓度组合是:MS+0.5mg/L TDZ+2.0mg/L NAA,其诱导体细胞胚胎的发生率为77.14%;采自于甘肃舟曲县立节镇的宜昌百合体细胞胚胎发生的诱导最佳激素浓度组合是:MS+0.5mg/L TDZ+1.5mg/L NAA,其诱导体细胞胚胎的发生率为85.71%。暗培养是诱导野生宜昌百合的体细胞胚胎发生的有利条件。鳞片的不同部位诱导体细胞胚胎发生、分化的能力由高到低排列顺序为:基部>中部>上部。60g/L的蔗糖浓度为诱导宜昌百合体细胞胚胎发生的最佳蔗糖浓度,其诱导率最高是90%。0.1g/L活性炭浓度为诱导宜昌百合体细胞胚胎发生的最佳活性炭浓度,其诱导率最高是83.33%。3.野生宜昌百合快繁与再生体系建立的最佳途径的研究:野生宜昌百合的一个接种鳞片通过不定芽诱导方式平均生成4.3个小鳞茎,一个接种鳞片通过体细胞胚诱导方式平均生成11.5个小鳞茎。宜昌百合的体细胞胚诱导方式是宜昌百合快繁和再生体系建立的更好的途径。4.野生宜昌百合试管苗的膨大、生根以及移栽的研究:影响野生宜昌百合试管苗鳞茎膨大的最佳培养基是:2MS+0.2mg/L NAA,增大倍数最大为2.51倍。野生宜昌百合生根的最佳培养基为:MS+0.2mg/L NAA+0.1g/L AC,最长根达到35.74cm。与此同时,本研究结果表明暗培养也是影响野生宜昌百合试管苗鳞茎膨大及生根的有利条件。野生宜昌百合试管苗的最佳炼苗时间为8天,移栽后成活率最高为83%;1/2MS营养液喷施移栽植株的叶面能达到最佳的效果,成活率达到80%;鳞茎直径大于1cm时移栽苗成活率最高,达到了88%;试管苗的根长在1-2cm时,移栽苗的成活率最高为77%。更多还原

【Abstract】 ‘Qinba’Mountains is one of areas with the most plentiful wild lily resources,the area has 11 kinds of wild lily Including variety.In recent years, the environment of the wild lily resources in‘Qinba’Mountains has been destroyed by human factors. Population propagation is also infiltrated by external plant species and threated by commercial ball invading.The work of protecting and preservating wild lily resources is very critical. Somatic embryogenesis is one of the in vitro culture of lily methods,which have many advantages such as large quantities,high speed,complete structure and high regeneration rate.This experiment studied tissue culture systerm and somatic embryogenesis systerm of wild lily in‘Qinba’Mountains,and researched the factors affected the somatic embryogenesis induced. The main results in this study as follows:1. The experiment of in vitro shoot induction of Lilium leucanthum:The best sterilization time of scales of Lilium leucanthum is: the combination is Alcohol 30s and HgCl2 10min. In the process of disinfection,the pollution rate is 33.33% and the mortality rate is 10%. the best hormonal combination inducing adventitious bud is: MS+1.0mg/L6-BA+0.3mg/LNAA,the induction rate is 86.67%. The ability of different locations in inducing in vitro shoot is :middle>outer>inner.2. The experiment of induction somatic embryogenesis of Lilium leucanthum:The best hormone combination in inducing somatic embryogenesis of three different ecotype of Lilium leucanthum is different. The best hormone combination of the Lilium leucanthum in YuDu,ZhenBa,ShaanXi is: MS+0.5 mg/L TDZ+1.0 mg/L NAA, the induction rate is91.43%. The best hormone combination of the Lilium leucanthum in LiuHou,LiuBa,ShaanXi is: MS+0.5 mg/L TDZ+2.0 mg/LNAA, the induction rate is77.14%. The best hormone combination of the Lilium leucanthum in LiJie,ZhouQu,ShaanXi is: MS+0.5 mg/L TDZ+1.5 mg/LNAA, the induction rate is 85.71%.Dark condition is favorable for inducing somatic embryogenesis of Lilium leucanthum.The ability of different position of scales in inducing somatic embryogenesis of Lilium leucanthum is: down>middle>upper. When sugar concentration of medium is 60g/L, the induction rate is 90% which is the highest.When activated carbon concentration of medium is 0.1 g/L, the induction rate is83.33%which is the highest.3. The best way to establish rapid propagation and regeneration system of Lilium leucanthum:The average number of new bulblet generating by the way of inducing adventitious buds is 4.3. The average number of new bulblet generating by the way of inducing somatic embryogenesis is 11.5.the way of somatic embryogenesis induced is the better way of rapid propagation and regeneration system of Lilium leucanthum:4. The experiment of enlargement、taking root、transplanting of tube plantlet of Lilium leucanthum:The best t hormone combination in enlarging tube plantlet of Lilium leucanthum is : 2MS+0.2 mg/LNAA,Dark condition is favorable for enlarging tube plantlet of Lilium leucanthum. The best hormone combination in taking root of tube plantlet of Lilium leucanthum is: MS+0.2 mg/L NAA+0.1g/LAC,the longest roots is 35.74mm. Dark condition is also favorable for taking root of tube plantlet of Lilium leucanthum.The best acclimatization time of the tube plantlet is 8 days. The survival rate is 83% which is the highest.1/2MS is the best nutrient solution,the survival rate is 80%.When the diameter of bulb is greater than 1cm, The survival rate is highest which is 88%.When the length of root of tube plantlet is between 1cm and 2cm,the survival rate is highest which is 77%.更多还原