山桐子繁殖及组织培养技术研究
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摘要
山桐子(Idesia polycarpa Maxim.)为大风子科山桐子属植物,是重要的木本油料、用材及观赏树种,具有很高的开发利用价值。研究山桐子繁殖及组织培养技术,对其资源开发和遗传改良具有重要意义。本文围绕山桐子繁殖及组织培养技术开展了种子催芽、根段繁殖和组织培养等方面的研究工作。主要研究结果如下:
1.以山桐子种子为材料,研究了浸种时间、浸种温度、催芽温度和GA3处理对其萌发的影响。结果表明:山桐子种子最适种子浸种温度为30℃、最适浸种时间为24h、最适催芽温度为25℃、GA3最适处理浓度为200mg/L,在最适组合条件下其发芽率、发芽势和活力指数可分别达到81.00%、63.67%和5.02。
2.以山桐子根段为材料,研究了基质、母树年龄、根段长度、根段粗度及激素种类等因素对山桐子根段繁殖的影响。结果表明:以1:1的河沙与珍珠岩为基质,以3年生苗木10cm长、1.0cm粗的根段为繁殖材料,用100mg/LABT生根粉浸泡30min,可以获得较为理想的结果,根段萌芽率可达92.0%。
3.以山桐子种子和叶片为外植体,研究了外植体种类、基本培养基和植物激素(生长调节物质)等因素对山桐子组织培养各个环节的影响,建立了山桐子高频组织培养再生体系。结果表明:种子为诱导愈伤组织的最佳外植体,先用70%酒精消毒2min,再用0.1%HgCl2消毒10min是最适宜的消毒方法,成活率达95.0%;种子诱导愈伤的最适培养基是MS+6-BA1.0mg/L+NAA1.0mg/L+3%蔗糖+0.6%琼脂,诱导率达96.8%,愈伤组织生长良好;愈伤组织不定芽分化的最适培养基是MS+ TDZ0.08mg/L+6-BA 1.5mg/L+NAA0.05mg/L+3%蔗糖+0.6%琼脂,诱导率达48.3%以上;腋芽增殖的最适培养基是1/2MS+TDZ0.03mg/L+6-BA2.0mg/L+NAA0.1mg/L+3%蔗糖+0.6%琼脂,增殖系数为4.83;不定芽生根的最适培养基是1/2MS+IBA 0.5mg/L+2%蔗糖+0.6%琼脂,培养40d后生根率可达88.0%。
Idesia polycarpa, as an energy, ornament and timber using tree species, has high value for application. Obtaining some techniques of propagation and tissue culture has great theoretical and realistic significance for resources exploitation and genetic improvement of this species. The technique of improving seed germination, tissue culture and root segment propagation of Idesia polycarpa were studied in this thesis. The chief results are as follows:
Using the seeds of Idesia polycarpa as materials, the effects of different treatment methods, including soaking time, soaking temperature, incubating temperature and concentration of GA3, on germination were studied. The results showed that the suitable treatment method was incubating under 25℃condition after soaking with 200mg/L GA3 for 8h and water for 16h at 30℃. Under which the germination rate, germination energy and vigor index were 81.00%, 63.67% and 5.02 respectively.
The root segments propagation technique of Idesia polycarpa was also explored in this study by testing various factors including cutting media, ages of the mother trees, length and diameter of root segments and hormone types. In the media of 1:1 sand and perlite, the survival rate of root segments from 3 year old trees with 10cm in length and 1.0cm in diamete, after soaking in 100mg/LABT for 30min, could reached 92.0%.
Using seeds and tender leaves of Idesia polycarpa as explants, the impact of explants, base mediums and plant hormones (or plant growth regulator) on tissue culture of Idesia polycarpa were studied. A tissue culture system with higher regeneration ratio of the species was established. Seeds were the suitable explants for inducing calli. The best process for seeds sterilize was dipping in 70% alcohol for 2min and then in 0.1%HgCI2 for 10min.The survival rate was 95%. On the medium of MS + NAA 1.0mg/L +6-BA 1.0mg/L + 3% sucrose + 0.6% agar, calli could beinduced successfully from explants with a rate of 96.8%. The optimum medium for inducing adventitious buds was MS + TDZ 0.08mg/L + 6-BA 1.5mg/L+ NAA 0.05mg/L + 3%sucrose + 0.6%agar, and the induction rate was more than 48.3%. Axillary buds chould be induced on the 1/2MS medium with 0.03mg/LTDZ, 2.0mg/L6-BA, 0.1mg/L NAA, 3%sucrose and 0.6%agar, and the proliferation rate reached 4.83.The elongated axillary buds were rooted at a rate of 88.0% on 1/2MS medium with 0.5mg/L IBA, 2%sucrose and 0.6% agar after culturing 40d.
1.以山桐子种子为材料,研究了浸种时间、浸种温度、催芽温度和GA3处理对其萌发的影响。结果表明:山桐子种子最适种子浸种温度为30℃、最适浸种时间为24h、最适催芽温度为25℃、GA3最适处理浓度为200mg/L,在最适组合条件下其发芽率、发芽势和活力指数可分别达到81.00%、63.67%和5.02。
2.以山桐子根段为材料,研究了基质、母树年龄、根段长度、根段粗度及激素种类等因素对山桐子根段繁殖的影响。结果表明:以1:1的河沙与珍珠岩为基质,以3年生苗木10cm长、1.0cm粗的根段为繁殖材料,用100mg/LABT生根粉浸泡30min,可以获得较为理想的结果,根段萌芽率可达92.0%。
3.以山桐子种子和叶片为外植体,研究了外植体种类、基本培养基和植物激素(生长调节物质)等因素对山桐子组织培养各个环节的影响,建立了山桐子高频组织培养再生体系。结果表明:种子为诱导愈伤组织的最佳外植体,先用70%酒精消毒2min,再用0.1%HgCl2消毒10min是最适宜的消毒方法,成活率达95.0%;种子诱导愈伤的最适培养基是MS+6-BA1.0mg/L+NAA1.0mg/L+3%蔗糖+0.6%琼脂,诱导率达96.8%,愈伤组织生长良好;愈伤组织不定芽分化的最适培养基是MS+ TDZ0.08mg/L+6-BA 1.5mg/L+NAA0.05mg/L+3%蔗糖+0.6%琼脂,诱导率达48.3%以上;腋芽增殖的最适培养基是1/2MS+TDZ0.03mg/L+6-BA2.0mg/L+NAA0.1mg/L+3%蔗糖+0.6%琼脂,增殖系数为4.83;不定芽生根的最适培养基是1/2MS+IBA 0.5mg/L+2%蔗糖+0.6%琼脂,培养40d后生根率可达88.0%。
Idesia polycarpa, as an energy, ornament and timber using tree species, has high value for application. Obtaining some techniques of propagation and tissue culture has great theoretical and realistic significance for resources exploitation and genetic improvement of this species. The technique of improving seed germination, tissue culture and root segment propagation of Idesia polycarpa were studied in this thesis. The chief results are as follows:
Using the seeds of Idesia polycarpa as materials, the effects of different treatment methods, including soaking time, soaking temperature, incubating temperature and concentration of GA3, on germination were studied. The results showed that the suitable treatment method was incubating under 25℃condition after soaking with 200mg/L GA3 for 8h and water for 16h at 30℃. Under which the germination rate, germination energy and vigor index were 81.00%, 63.67% and 5.02 respectively.
The root segments propagation technique of Idesia polycarpa was also explored in this study by testing various factors including cutting media, ages of the mother trees, length and diameter of root segments and hormone types. In the media of 1:1 sand and perlite, the survival rate of root segments from 3 year old trees with 10cm in length and 1.0cm in diamete, after soaking in 100mg/LABT for 30min, could reached 92.0%.
Using seeds and tender leaves of Idesia polycarpa as explants, the impact of explants, base mediums and plant hormones (or plant growth regulator) on tissue culture of Idesia polycarpa were studied. A tissue culture system with higher regeneration ratio of the species was established. Seeds were the suitable explants for inducing calli. The best process for seeds sterilize was dipping in 70% alcohol for 2min and then in 0.1%HgCI2 for 10min.The survival rate was 95%. On the medium of MS + NAA 1.0mg/L +6-BA 1.0mg/L + 3% sucrose + 0.6% agar, calli could beinduced successfully from explants with a rate of 96.8%. The optimum medium for inducing adventitious buds was MS + TDZ 0.08mg/L + 6-BA 1.5mg/L+ NAA 0.05mg/L + 3%sucrose + 0.6%agar, and the induction rate was more than 48.3%. Axillary buds chould be induced on the 1/2MS medium with 0.03mg/LTDZ, 2.0mg/L6-BA, 0.1mg/L NAA, 3%sucrose and 0.6%agar, and the proliferation rate reached 4.83.The elongated axillary buds were rooted at a rate of 88.0% on 1/2MS medium with 0.5mg/L IBA, 2%sucrose and 0.6% agar after culturing 40d.
引文
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