【作者】 谢艳平；

【导师】 吴江生；

【作者基本信息】 华中农业大学 ， 作物遗传育种， 2008， 硕士

【摘要】 甲基磺酸乙酯(Ethl methane sulfonste,EMS)是一种能够诱导染色体结构变异的烷化剂,已被广泛应用于诱变育种和种质创新。本实验采用不同EMS浓度(0.02%、0.04%、0.08%(v/v))和处理时间(10min和20min)对7679-1、7679-2、7680、7681和2145在内的5个品系的甘蓝型油菜小孢子进行诱变处理,对诱变后小孢子的胚胎发生能力、植株再生能力、再生植株形态和种子品质变异等进行了研究。主要研究结果如下:1.不同基因型小孢子胚胎发生能力均存在明显差异。品系7679胚胎发生能力达到100%,2050-2等材料胚胎发生能力低,产胚率0—1.7个/蕾。同一品系不同株系之间胚胎发生能力无显著差异,如品系2050的4个株系的单株胚胎发生能力在0—4.0%之间。2.不同EMS浓度和处理时间对小孢子产胚率的影响。在EMS浓度相同时,材料7679-2、7680小孢子产胚数随诱变时间延长而减少;在处理时间相同时,材料7679-2、7680小孢子产胚数随诱变剂浓度增加而减少,且随着诱变剂浓度增大产胚数下降速度加快。3.确定不同基因型材料的EMS诱变半致死剂量。7679-1处理10min时的半致死剂量为0.04%;7679-2处理10-20 min时的半致死剂量为0.08%;7680处理20min时的半致死剂量0.04%。4.获得了不同表型的突变体。经济性状突变:矮化、短角果、分枝数增多等突变体;花器官突变:粉色花瓣、白色花瓣、雄蕊多于6枚、花瓣数增加、柱头外露等突变体;种子品质突变:获得了含油量43.81%的植株(供体植株含油量38.12-40.93%),油酸含量68.64%的DH植株(供体植株油酸含量62.93-65.82%)。。5.EMS诱变对小孢子染色体加倍的影响。7679-2未诱变处理的小孢子经秋水仙碱处理后再生植株加倍率54.0%,EMS诱变后经秋水仙碱处理再生植株加倍率在50.0-92.4%,大部分高于未诱变处理;7680未诱变处理的小孢子经秋水仙碱处理的加倍率96.7%,EMS诱变后经秋水仙碱处理再生植株加倍率在85.7-98.0%,EMS诱变对再生植株染色体加倍效果有影响。更多还原

【Abstract】 Ethl methane sulfonste(EMS) is a kind of alkylating agent which has remarkable mutagenesis effect in chromosome mutagenesis. It has been widely applied in auxiliary breeding and germplasm innovation. This experiment adopts three EMS concentrationsnamely 0.02%, 0.04% and 0.08% (v/v) and two treating durations which are 10 minutes and 20 minutes to treat on in vitro microspore of nine lines from five strains in Brassica napus ,that including 7679-1、7679-2、7680. Investigate the microspore’s embryogenesis ,regenerative ability, and the microspore-derived plant’s phenotypic and qualitive variation. The finding results are listed as follows:1. There are diffences in microspore embryogenesis of various genotypes, material7679 get embryo easily,the ability of embryogenesis can reach 100%. 2050-2 embryogenesis is very low, which produces 0-1.7 embryoes per alabastrum. Embryogenesis ability tends to be consistent in different lines of the same strain. For example the embryogenesis ability are all extremely low as 0-4.0% in fore lines of strain 2050.2. The influence of EMS concentration and treating duration on microspore-yield embryo number . when EMS concentration is same every material’s embryo number are always reduced by prolong of treatment duration and the longer the time is the speed ofembryo generation descends slower..when treating duration is same 7679-2、7680’sembryo number reduced accelerative by increase EMS concentration, and the more the concentration is the speed of embryo generation descends faster..3. Every material has its own medial lethal concentration(LC5o): 7679-1 producing embryoes was about half of the CK when EMS concentration was 0.04% and the duration of EMS treatment was 10min; 7679-2 producing embryoes was about half of the CK when EMS concentration was 0.08% and the duration of EMS treatment was 10-20min;7680 producing embryoes was about half of the CK when EMS concentration was 0.04% and the duration of EMS treatment was 20min. 4. The phenotypic variation has get from DH plants’ by EMS mutagenesis. Variation in economic character : height decreases, silique becomes shorter , branching number increases. Flower organ morphological variations including : pink petal, white peta; got more than six stamens, more than fore petalsl, and stigma exsertion. Qualitive variation:got a DH plant whose oil content achieved 43.81% (Donor adult plant index of oil 38.12-40.93%), a DH plant whose oleic acid content was 68.64% (Donor adult plant oleic acid content 62.93-65.82%).5. EMS treatment influence the chromosome double rate of microspore-derived plants. In 7679-2’s regeneration group, CK chromosome double rate was 54.0% by treating with colchicines, regeneration groups which treated by EMS before colchicines treating that chromosome double rate were 50.0-92.4%, were mostly higher than CK. In 7680’s regeneration group, CK chromosome double rate was 96.7% by treating with colchicines; regeneration group which treated by EMS before colchicines treating that chromosome double rate were 85.7-98.0%. In a certain amount of concentration, EMS mutagenesis treatment approved the double of regeneration plant chromosome.更多还原