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树上干杏再生体系的建立与多倍体诱导
周黎
学位类型硕士
导师王晓军
2011-05-25
学位授予单位中国科学院研究生院
学位授予地点北京
学位专业有机化学
关键词 组织培养 植株再生 秋水仙素 多倍体诱导
摘要论文以新疆树上干杏种子和茎段为材料,系统研究了其组织培养与再生体系建立的过程,对各个培养阶段的主要影响因素进行了探讨,并采用化学方法对其进行多倍体诱导,为新疆杏品种开展生物技术育种奠定了基础,现得结果如下: 关于树上干杏再生体系建立的研究:种子消毒处理以75%乙醇溶液处理30s,0.1%升汞溶液处理30s, 15%双氧水溶液处理20min,灭菌无菌水冲洗6次效果较好,平均发芽率和污染率分别为87.5%和12.5%。茎段消毒处理为75% 乙醇溶液处理30s,0.1% 升汞溶液处理5 min,15%双氧水溶液处理20min,灭菌无菌水冲洗6次效果较好,发芽率和污染率分别为89.6%和8.9%。茎段的取材以当年春季新抽出的枝条最好,发芽率高污染率低。以茎段为材料诱导不定芽最适培养基:改良MS+1.0 mg/L TDZ+0.1 mg/L NAA,出芽率可达90.8%。不定芽增殖培养的最佳培养基:QL’+0.4 mg/L6-BA+0.04 mg/LIBA,增殖倍数可达5.2,新生苗平均高度为3.1cm。诱导生根的最佳培养基为:1/2DKW+0.4mg/LIBA。接种后暗培养12d生根效果最好,生根率达到83.3%,平均每株生根数目为4条,生根指数为499.8。 关于多倍体诱导的研究:茎段浸泡法诱导杏多倍体,以0.4%秋水仙素溶液浸泡1d得到的变异率最高,可达15.8,再生率为15.8。茎段混培法诱导杏多倍体,以在含0.2%秋水仙素的培养基里培养10d得到最多变异株,变异率为20%。种子浸泡法诱导杏多倍体,以0.5%秋水仙素溶液处理24h效果比较好,其发芽率和变异率分别是35%和15%。茎尖涂抹法诱导杏多倍体,以0.3%的秋水仙素溶液对茎尖处理的效果最好,存活率为71.4%,变异率为21.4%。变异株在诱导初期与对照株形态学上存在差异,如叶片面积变小,叶片长宽比由1.58变为1.14,节间长度由2.49cm变为1.58cm。变异株与对照株的气孔对比,气孔增大,密度减小。变异株与对照株气孔大小分别为27.30µm×19.55µm和17.20µm×13.85µm(长×宽),气孔密度分别为62.0 mm-2和109.0 mm-2。5颗变异株经流式细胞仪检测,最终确定有2株四倍体植株,1株嵌合体植株,2株二倍体植株。
其他摘要The tissue culture and regeneration system were systematically studied by using seeds and stems of apricot as meterial. And the influence of the whole process was researched detailedly, which laid the foundation for the transgenation and polyploidy breeding. Meanwhile, ployploid induction of apricot was studied primarily.The results were as follow: The results about regeneration system research: The most suitable disinfection method for apricot’ seeds was 75% C2H5OH treated 30s, 0.1% HgC12 treated 30s, 15%H2O2 treated 20min, and washed by germfree water 6 times. The rate of germination was 87.5%, the rate of pollution was 12.5%.The most suitable disinfection method for apricot’ stems was 75% C2H5OH treated 30s, 0.1% HgC12 treated 5min, 15%H2O2 treated 20min, and washed by germfree water 6 times. The rate of germination was 89.6%, the rate of pollution was 8.9%. we’d better take the explants in spring.The optimum medium for shoot induction was modified MS+1.0 mg/L TDZ+0.1 mg/L NAA.The regeneration rate was 90.8%.The optimum medium for shoot proliferation was QL’+0.4 mg/L6-BA+0.04 mg/LIBA. The proliferation coefficient was 5.2 and the average heigh of shoots was 3.1cm.The optimum medium for shoot rooting was 1/2DKW+0.4mg/LIBA. The best result was treated 12 days in dark , and the rooting rate was 83.3%, the average number of rooting is 4. The results about ployploid induction: Immersing shoots in 0.4% colchicine solution for 24 h induced a high number of autotetraploid plants. The induction rate reached as high as 15.8% and the regeneration rate of treated stems was 15.8.Stoots treated in proliferation medium supplied with colchicine .The best result was culture 10d in 0.2% colchicine medium and the induction rate was 20%.Seeds were immersed in various concentrations of colchicine solution. The best result was 0.5% colchicine treated 24h. The germination rate and the induction rate was 35% and 15%, respectively.Colchicine was applied directly to the apical growing tips. That 0.3% colchicine processed 3d achieve the best effectiveness. The survival rate and the induction rate was 71.4% and 21.4%, respectively.The differences of morphological and anatomical between diploid and tetraploid are obvious, the leaf index from 1.58 change to 1.14, internodes from 2.49cm change to 1.58cm. The average size of stomatas in diploid and tetraploid is 17.20µm×13.85µm and 27.30µm×19.55µm. The average density of stomatas in diploid and tetraploid is 109.0 mm-2 and 62.0 mm-2. Five variant plants were tested by FCM, two were teraploid , one was mixoploid and two were diploid.
文献类型学位论文
条目标识符http://ir.xjipc.cas.cn/handle/365002/4434
专题资源化学研究室
作者单位中国科学院新疆理化技术研究所
推荐引用方式
GB/T 7714
周黎. 树上干杏再生体系的建立与多倍体诱导[D]. 北京. 中国科学院研究生院,2011.
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