大果木莲种子生物学与离体繁殖技术研究
潘睿
学位类型硕士
导师孙卫邦
2010-05
学位授予单位中国科学院研究生院
学位专业植物学
摘要1. 大果木莲种子的休眠类型: 大果木莲种子种子透水性良好,胚很小,已分化但发育不完全;在25℃下30 d的萌发率只有2%,具有形态生理休眠特性。大果木莲种子在25/15℃下萌发率达到了82%,其种胚并不要求0~10℃的层积处理来打破其形态生理休眠且只有在临界萌发时才有明显生长。可以判定,大果木莲种子具有浅度简单型形态生理休眠,冷层积可以打破其休眠组成中的生理休眠。大果木莲种子在30/20℃,25/15℃,20/10℃下培养50 d时的萌发率较30 d时的萌发率有所扩展,20/10℃下GA3有效的打破了种子的生理休眠,由此大果木莲种子具有条件休眠的特性。经过不同时间的层积处理后,发现层积60 d是行之有效的打破大果木莲种子休眠的方法,而之后的层积处理使种子长期处于一个低温潮湿的环境,使种子进入次生休眠产生休眠循环。2. 有效打破大果木莲种子休眠及促进萌发的方法: 1000 mg·L-1的GA3在20~30℃下可以较为有效的打大果木莲种子的休眠。6-BA与KNO3对大果木莲种子的萌发没有显著的促进作用,但是这两种化学物质无论浓度的高低在20/10℃下都能够使大果木莲种子达到一个较高的萌发率且彼此之间的差异性并不显著。大果木莲的种子经过60 d的低温层积之后,其原有休眠几乎可以全部被打破。而在20/10℃下大果木莲种子的最高萌发率发生在层积40 d的处理下,层积了100 d的种子其萌发率也高达到82%。3. 大果木莲种子的储藏习性: 由耐干性测试所初步得出的大果木莲种子储藏习性为中间型。进一步试验中,大果木莲种子干燥至10~12%的含水量时,其活力均在90%以上。继续干燥至5%的含水量,大果木莲种子的生活力为87%,在3.7%的含水量时,仍有大部分种子存活。然后-20℃下密闭贮存三个月,其生活力只有30%(5%与4.7%含水量下种子活力综合考虑)左右,大多数种子失去了生活力。由此判断,大果木莲种子为中间型种子。4. 大果木莲种子的储藏条件: 大果木莲的种子可以忍耐4℃的低温,并长期保持种子的生活力,但对零下低温忍耐性很差,在10℃以上的温度下活力随含水量的降低而丧失。由此,可以将大果木莲的种子直接密闭封藏在4℃下或者进行适度的干燥至10%左右于-20℃储藏。低温湿藏60 d与120 d后种子的生活均在90%以上,可以看出,低温湿藏比干藏的效果要好的多而且操作简单便捷。5. 大果木莲的离体培养: 以大果木莲种子在]无菌体条件下萌发的小植株(未长出真叶)进行不定芽的诱导、增殖及不定根的诱导培养。将未小植株置于H+6-BA0.25mg·L-1(单位下同)+NAA0.125+KT0.1+IAA0.05的培养基上培养,可以诱导出多个诱导芽。诱导芽在H+6-BA0.5+KT0.1上培养,其增殖系数可达3.5,且生长健壮。在培养过程当中,诱导的不定芽会出现褐化,通过抗氧化剂的使用与切除得到了有效的缓解。
资助项目1. Seed dormancy type ofManglietia grandis seeds, The seeds of Manglietia grandis had good water-absorption and the embryos were underdeveloped. Fresh seeds only germinated to 2% at 25 ℃, so the seeds were morphophysiological dormant. The seeds germinated to 82% at 25/15 ℃,the embryos do not need 0~10 ℃ stratification to break the morphophysiological dormancy and only grew obviously when they were nearly germinated. So we can say that the seeds of Manglietia grandis have non-deep morphophysiological simple dormancy, and the cold stratification can break the physical dormancy component of there complex dormancies. The 50 days germination rate of Manglietia grandis seeds was higher than the 30 day germination rate at 30/20℃,25/15℃,20/10℃, and the GA3 had broken the seeds physical dormancy effectively, so the Manglietia grandis seeds were conditional dormant. After different time cold stratifications, we found that the 60 days stratification was the most effective way to break the dormancy. The others’ stratifications made a long time and moist environment, so the seeds would have a secondary dormancy, entering into the dormancy circle. 2. The way to break the dormancy of Manglietia grandis seeds effectively and to improve the germination, 1000 mg·L-1 GA3 could break the dormancy of Manglietia grandis seeds at 20~30℃ somehow. 6-BA and KNO3 couldn’t improve germination effectively, but these two chemicals could make a considerably high germination at 20/10℃ no matter with the concentration. And the differences of each other were not remarkable. The dormancy of 60 days cold stratification Manglietia grandis seeds almost could been broken. But at 20/10℃ the highest germination happened at the 40 days stratification, the 100 days stratification’ s germination also high to 82%. 3. The storage behavior of Manglietia grandis seeds, After the desiccation tolerance test, we can know generally that the storage behavior of Manglietia grandis seeds are intermediate type. In the further experiment, when the seeds were dehydrated to 10~12% water content, the viability was above 90%. Go on the dehydration to 5% water content, the seeds viability was 87%,when the water content was 3.7%, there still were considerable live seeds. Then the seeds were air tightly stored at -20℃ for 3 months, the viability was only about 30%(comprehend the viability of 5% and 4.7% water content together), most seeds lost their viability. So the Manglietia grandis seeds were intermediate seeds. 4. The storage condition of Manglietia grandis seeds, The Manglietia grandis seeds can endure 4℃ low temperature, and keep the viability for quite a long time, but can’t endure the sub-zero low temperature. At the 10℃ above temperature, the seeds viability went down as the water content lost. So we can airtightly store the Manglietia grandis seeds directly at 4℃ or make proper dehydration to about 10% water content then store at -20℃. After low temperature moist storage 60 days and 120 days, the seeds viability were all above 90%, we can concluded that the low temperature moist storage was more effective than the dry storage, and easy to manipulate. 5. The in vitro culture of Manglietia grandis seeds, The in vitro culture experiment used seedlings of Manglietia grandis as the explants. Put the seedlings which only have the cotyledons into the culture medium of H+6-BA0.25+NAA0.125+KT0.1+IAA0.05 mg·L-1(the fallowing units are as the same), several adventitious buds had be induced out. Put all these adventitious buds into the medium of H+6-BA0.5+ KT0.1, the multiplication coefficient is 3.5, and all of the buds grew well. It’s not hard to induce out the adventitious buds and make proliferations, the problem was that during the process there had be browning effects on the seedlings and buds, the problem was settled by cutting and using the antioxidants
语种中文
文献类型学位论文
条目标识符http://ir.kib.ac.cn/handle/151853/16064
专题昆明植物所硕博研究生毕业学位论文
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潘睿. 大果木莲种子生物学与离体繁殖技术研究[D]. 中国科学院研究生院,2010.
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