【作者】 何志祥；

【导师】 谭晓风； 曹孟良；

【作者基本信息】 中南林业科技大学 ， 林学， 2005， 硕士

【摘要】 本文对梨的组织培养技术进行了较为系统的研究。分别以13个杂交或自交梨组合的胚、翠冠梨的茎段、新高梨自交组培苗叶片为材料，实验研究了其相应的离体再生方式、培养条件及其关键影响因子，为梨优良种质的获得及其良种快速繁殖提供了科学依据和技术基础。主要结果如下： 1、胚培养。以13个砂梨和白梨系统的杂交和自交组合为研究对象，研究了幼胚组织培养技术，并成功地获得了再生植株。结果表明，以培养基1／2MS+6-BA0.5mg／L适于幼胚再生，但不同结合之间、尤其是杂交与自交组合之间，幼胚的离体再生能力和表现有很大差异，在该培养基配比条件下，砂梨与砂梨杂交的组合，无论哪种梨品种为父本，幼胚出芽率和生根率都较其他组合好。外植体对胚培养有影响，带部分子叶的胚比不带子叶或带完整子叶的胚更适于该培养条件。 2、茎段培养。在激素水平为6-BA2.0mg／L和IBA0.2mg／L的条件下，选取1／2MS为基本培养基可以获得高于以WPM或AS为基本培养基的外植体再生率。有芽茎段和无芽茎段具有不同的再生方式及不同的适宜条件。有芽茎段倾向于不经过脱分化而直接出芽，可以省略诱导愈伤阶段；而无芽茎段培养则必须经过脱分化阶段，先诱导出愈伤组织，然后诱导不定芽。就本研究所选用的3种基本培养基而言，AS较适合无芽茎段的愈伤组织诱导，愈伤组织生长良好。 3、叶片培养。经过正交实验筛选出最佳培养基配比为NN69+0.5 mg／LTDZ+1.0 mg／L 6-BA+0.3 mg／L IAA。接种方式以叶基部向上的叶片愈伤率和出芽率都是最高的。叶龄对叶片培养有较大影响，以胚培养再生后继代培养1 5-30d的叶片进行叶培养，无论是愈伤率还是出芽率，都明显优于继代培养30d以上的叶片。 4、移栽。移栽基质和移栽方式对组培苗移栽成活率无明显影响，不管在哪种基质用哪种方式盘炼，只要无菌苗出瓶时生长健壮均能获得较高的移栽成活率，这说明组织培养外植体成功与否主要在于无菌培养阶段，移栽基质和移栽方式只影响其适应外界条件的时间长短。更多还原

【Abstract】 The tissue culture of the pear has been researched systematically. Used the stems of Cuiguan pear, the embryo of 13 different combinations of pear hybridize and self-breeding leaves of Niitaka pear as explants, the basic conditions of culture and some essential factors of regeneration have been determined. In addition, the transplants of plantlet have been researched.All of these researches are benefit to the foundation of pear tissue culture and its implication on rapid propagation of fine varieties. The main results are shown as follows:1, embryo culture: With the medium of 1/2MS+6-BA0.5mg/L, the embryo are easy to regenerate plantlet. At the same medium,the different capabilities of regeneration have been observed on different combinations of pear hybridize. Compared to other combinations ,the combination of Pyrus pyrifolia and Pyrus pyrifolia have the higher buding and rooting frequency at the medium above. Moreover, the pear embryo with part of cotyledon can grow better than the pear embryo without cotyledon or the pear embryo with whole of cotyledon.2, stem culture: Under the hormone level of 6-BA2.0mg/Land IBA0.2mg/L, the regeneration ratio on the basic medium of 1/2 MS is higher than on the basic medium WPM or AS . The stem with buds can regenerate plantlet directly without dedifferentiation, but the stem without buds must be induced callus firstly. According to the three basic medium of this research, AS adapts to the no bud stem culture, and the callus are health, but it need look for another dividing medium.3, leaf culture:After orthorhombic test, the NN69+0.5 mg/1 TDZ+0.3 mg/1 IAA has been determined as the best medium. In addition, it can obtain the highest callus frequency and budding frequency with inverted explanting of leaves. Subctltured different ages of leaves regenerated from embryo culture,both callus frequency and budding frequency of 15-30-day-old are much higher than 30-day-old4, transplant: the technique of transplant has been researched. The result shew that the plantlets grew well no matter what transplant method. The result indicated that the important stage which determine tissue culture whether or nor success is at tube-stage, and the transplant methods only affects the long of plantlets to adapt to growth condition.更多还原