【作者】 孙珍夏；

【导师】 彭定祥；

【作者基本信息】 华中农业大学 ， 作物栽培学与耕作学， 2007， 硕士

【摘要】 苎麻(Bochmeria nivea L．Gaud)是我国特产的重要纤维作物。占全世界总产量的95％，纺织加工能力约占80％。其纤维品质优良，在纺织上有重要地位，因而在世界上素有“中国草”的美誉。但在生产上存在遗传背景复杂，育种周期太长的问题，严重限制了苎麻新品种的培育。目前已经有许多作物通过单倍体途径获得纯和二倍体植株用于育种研究。花药培养是获得单倍体比较简单的方法。本实验通过对谈家何麻等13个品种和材料花药培养愈伤诱导频率主要影响因素的研究，初步形成了一套从苎麻花药愈伤诱导到愈伤倍性鉴定的完整体系，为下一步研究打好了基础。主要研究内容和结果如下：在外植体污染控制方面，本研究认为使用含有效氯为1％的次氯酸钠溶液灭菌效果好而且对外植体的毒害作用比较小。低温预处理会增加苎麻花药的污染频率，液体培养基要比固体培养基更容易污染。本实验通过对基本培养基、蔗糖浓度、激素配比、基因型的因素的筛选对苎麻花药培养愈伤诱导体系进行了优化。MSN、B5N这两个基本培养基的诱导效果比较好，蔗糖浓度在3％的时候诱导率最高，采用2，4-D2.0mg／L附加BA0.2 mg／L的激素组合对苎麻花药愈伤诱导效果最稳定。并且筛选到了一批诱导率比较高的材料，其中94-10的诱导率最高，达到81.05％。愈伤组织的分化与增殖实验中，以谈家何麻、白里子青、94-10的花药愈伤转入含有不同激素配比的分化培养基，但均未发现有芽的分化。愈伤在2，4-D0.5mg／L附加BA0.5mg／L的培养基上生长缓慢，结构为松散的黄色颗粒。对谈家何麻及白里子青的花药愈伤及由愈伤诱导产生的根，分别进行流式细胞仪和细胞学鉴定，但两种方法均未发现有单倍体细胞。更多还原

【Abstract】 Ramie (Bochmeria nivea L. Gaud.) is an important fiber crops and speciallyproduction in China. The ramie output in China accounts for the world ultimate output95％, the textile processing ability approximately composes 80％. Because of its excellenttextile fiber quality, ramie has the important status in textile industry. Thus ramie has thefine reputation of "China Grass" in the world.But in the application, the questions of its extremely complex heredity backgroundand long traditional breeding method cycle, have seriously limited the scientific researchdevelopment of ramie. Many crops have been obtained dihaploid plant by double thehaploid in present. The anther culture is a simple method to obtain haploid. We haveestablished the complete protocol for ramie anther callus induction and callus ploidyidentify by researching the major effect factors of the anther callus induction frequency in13 ramie varieties or materials. And it was the foundation for developing research. Themain research content and result as follows:The research of anther contaminated factors indicated that it was easy to becontaminated whenthe anther was kept in 4℃for longer than 2 days or cultured inliquid medium. The callus induction frequency of Tanjiahema, Jiangkou, Huangganma,Cuba ramie were highest when sterilized by 1％NaClO solution.We have optimized the ramie anther culture method by selecting the basic media,sucrose concentration, plant growth regulators and genotypes. Anthers cultured in MSNand B5N media indicated higher callus induction frequency than in other media. Whenthe media contained 3％sucrose concentration the anthers had high callus inductionfrequency. 9 media which contained different plant growth regulators and concentrationswere selected. The result shows that major part of the media can induct anther callus. Andthe medium which contains 2,4-D 2.0mg/L and BA 0.2 mg/L had stable effect of callusinduction. And we have selected some materials which have high callus inductionfrequency, such as 94-10 (81.05％). We translated the anther callus of Tanjiahema, Bailizi Qing, 94-10 to differentdifferentiation media. Callus grew slowly and their structure were loose decadent pelletwhen they cultured in the media contains 2,4-D 0.5mg/L and BA 0.5mg/LThe anther callus were identified the ploidy by FACSCalibur and cytology method.No haploid cells were found by the two methods.更多还原