百合组织培养快繁研究
摘要
本试验选用铁炮、粉香水两个百合品种的鳞片，帝伯、西伯利亚、铁炮、金百合四个百合品种的花器官为外植体进行培养，分别在无性培养系建立，胚性愈伤组织的增殖及分化，生根结鳞茎和移栽四个阶段进行了研究，得出以下结果：
1．无性培养系建立阶段
用铁炮和香水百合的鳞片作为外植体放在各种培养基中进行培养，对其各部位的污染率、诱导分化率等进行比较。试验结果表明，鳞茎中部和内部的鳞片污染率低，为较佳的外植体培养材料。同一片鳞片，基部最易分化，其次为中部，顶部较难分化。在所试验的培养基中，MS+BA2.0(单位为mg·L~(-1)下同)+NAA0.2培养基最适合铁炮百合鳞片的分化，分化率为83％，而粉香水百合的鳞片在MS+BA_(4.0)+NAA_(0.2)中的分化率最高，为75％。
用帝伯、西伯利亚、铁炮、金百合四个品种的花器官作为外植体建立无性培养系。试验结果表明，品种间的分化难易存在差异，从易到难的排序为帝伯、西伯利亚、铁炮、金百合，同一品种不同部位的分化难易也存在差异，从易到难的排序为花丝、花托、子房、花冠、花柱、花药。在所试的培养基中，MS+BA0.5+NAA0.2和MS+BA1.0+NAA0.5培养基的分化效果较好。
无菌苗叶片诱导胚性愈伤组织的的试验表明最适合培养部位为叶片的基部。基部培养较适合的培养基为MS+BA2.0+NAA0.2，MS+BA1.0+NAA0.2。
2．胚性愈伤组织增殖及分化阶段
在鳞茎胚性愈伤组织增殖及分化的试验中，对铁炮百合鳞片及无菌苗叶片较
适培养基为MS+BAZ .0+NAAO.2;对粉香水百合鳞茎较适合增殖的培养基为MS+
BAI .0，较适合分化的培养基为MS+BAO.5+NAAO.05;花器官(帝伯和西伯利亚)
的较适培养基为MS+BAI .0十NAAO.2，而铁炮百合为MS+B川.0+NAAO.5。
3.生根结鳞茎阶段
在生根结鳞茎培养中，激素的含量和种类、糖的浓度、大量元素的浓度等对
百合生根结鳞茎都有影响。MET(多效哩)有助于鳞茎的形成，较适浓度为2.0
mg.L一，和2.5 mg.L一，。NAA有利于根的生长，不利于鳞茎的形成。糖也有助于鳞茎
的形成，较适浓度为6Og.L一，。降低大量元素的浓度有利于生根结鳞茎，较适的浓
度为1/4MS。
4.移栽阶段
生根苗的移栽分四个步骤，沙床的准备、炼苗、假植和移栽大田或盆栽。各
种处理中移栽成活率最高可达到100%。
Bulblets of lily of two species Lilium longiflorum ,Oriental lily and floral organs of lily of four varieties - Diber, Siberia, Lilium Longiflorum and Lilium trompeten were selected for in vitro culture .Four stages of primary research were processed, that was, establishment of clone system, multiplication of embryonal callus, rooting and bulblet formation and transplantation. The result are shown as follows:
1. Stage of establishment of clone system
The bulblets of two species , Lilium Longiflorum and Oriental lily, were selected as explants for in vitro culture , and compared their contamination rate and inducing differentiation rate of different position in bulblet. The results of experiments indicated that the containminion rate of inner and middle part in bulblet were in low level. When different part in a single scale were cultured in the same medium the results showed that the basal part turned out to be the easiest part for inducing, the second was the middle part, the upper part was hard to induce. Among the inducing mediums in our experiment, MS medium supplemented with 2.0mg L-1 BA and 0.2 mg L-1 NAA was the best for Lilium Longiflorum , its inducing rate was 83 percent; while MS medium supplemented with 4.0mg L-1 BA and 0.2 mg L-1 NAA was suitable for Oriental lily, the inducing rate was 75 percent.
The floral organs of four varieties, Diber, Siberia, Lilium Longiflorum , and Lilium trompeten, were selected as explants for in vitro culture. It was founded that when they were culture in the same medium ,the inducing differentiation rate varied from varieties. Diber was the easiest variety for inducing which was followed by Siberia, Lilium Longiflorum, and Lilium trompeten. The inducing rates of different part in the same variety were also different. Their sequence, from easy to hard, was filament, floral receptable, ovary, corolla, style and anther. The explants were cultured in a series of inducing medium .The results showed that MS medium supplemented with 0.5mg L-1 BA and 0.5mg L-1 NAA and
MS medium supplemented with l.Omg'L"1 BA and O.Smg-L"1 NAA showed a better induction character.
When different part of bacteria and germ -free leafs were induced for embryonal callus, the basal part was the easiest to induce. MS medium supplemented with 2.0 mg.L"1 BA and 0.2mg.L"' NAA and MS medium contained l.Omg.L"1 BA and O.Zmg.L"1 NAA were suitable for basal part of leaf inducing culture.
2. Stage of multiplication and differentiation of embryonal callus
During the stage of embryonal callus of multiplication and differentiation ,it was found that, for Lilium Longiflorum bulblet and germ-free seeding , MS medium containing 0.2 mg.L"1 BA and 0.2 mg.L"1 NAA was the best medium; for Oriental lily, the best multiplication medium was MS supplemented with 1.0 mg.L"1 BA, and the best differentiation medium was MS supplemented with O.Smg.L"1 BA and 0.05 mg.L ' NAA; MS supplemented with l.Omg.L"1 BA and 0.2 mg.L"1 NAA were suitable for floral organs of Siberia and Diber ; and MS containing l.Omg.L"1 BAand O.Smg.L"1 NAA were fit to floral organs of Lilium Longiflorum .
3. Stage of rooting and bulblet formation
During the stage of rooting and bulblet formation, the result showed that the category and the concentration of hormone, the concentration of sugar and the concentration of macroelement, affected rooting and forming of bulblets. The application of PP333 benefited the bulblet formiation, the best concentration were 2.0 mg.L"1 and 2.5 mg.L"1. NAA promoted rooting but made against forming bulblet. Sucrose can help to form bulblet, the best concentration was 60g.L"1. It was favorable for rooting and bulblet formation in reducing the concentration of macroelement .1/4MS proved the best concentration.
4.Stage of transplantation
There were four stages in transplantation , that was sand bed prepairing , domestication, pseudoplantation and transplant soil and basin. The survival rate
1．无性培养系建立阶段
用铁炮和香水百合的鳞片作为外植体放在各种培养基中进行培养，对其各部位的污染率、诱导分化率等进行比较。试验结果表明，鳞茎中部和内部的鳞片污染率低，为较佳的外植体培养材料。同一片鳞片，基部最易分化，其次为中部，顶部较难分化。在所试验的培养基中，MS+BA2.0(单位为mg·L~(-1)下同)+NAA0.2培养基最适合铁炮百合鳞片的分化，分化率为83％，而粉香水百合的鳞片在MS+BA_(4.0)+NAA_(0.2)中的分化率最高，为75％。
用帝伯、西伯利亚、铁炮、金百合四个品种的花器官作为外植体建立无性培养系。试验结果表明，品种间的分化难易存在差异，从易到难的排序为帝伯、西伯利亚、铁炮、金百合，同一品种不同部位的分化难易也存在差异，从易到难的排序为花丝、花托、子房、花冠、花柱、花药。在所试的培养基中，MS+BA0.5+NAA0.2和MS+BA1.0+NAA0.5培养基的分化效果较好。
无菌苗叶片诱导胚性愈伤组织的的试验表明最适合培养部位为叶片的基部。基部培养较适合的培养基为MS+BA2.0+NAA0.2，MS+BA1.0+NAA0.2。
2．胚性愈伤组织增殖及分化阶段
在鳞茎胚性愈伤组织增殖及分化的试验中，对铁炮百合鳞片及无菌苗叶片较
适培养基为MS+BAZ .0+NAAO.2;对粉香水百合鳞茎较适合增殖的培养基为MS+
BAI .0，较适合分化的培养基为MS+BAO.5+NAAO.05;花器官(帝伯和西伯利亚)
的较适培养基为MS+BAI .0十NAAO.2，而铁炮百合为MS+B川.0+NAAO.5。
3.生根结鳞茎阶段
在生根结鳞茎培养中，激素的含量和种类、糖的浓度、大量元素的浓度等对
百合生根结鳞茎都有影响。MET(多效哩)有助于鳞茎的形成，较适浓度为2.0
mg.L一，和2.5 mg.L一，。NAA有利于根的生长，不利于鳞茎的形成。糖也有助于鳞茎
的形成，较适浓度为6Og.L一，。降低大量元素的浓度有利于生根结鳞茎，较适的浓
度为1/4MS。
4.移栽阶段
生根苗的移栽分四个步骤，沙床的准备、炼苗、假植和移栽大田或盆栽。各
种处理中移栽成活率最高可达到100%。
Bulblets of lily of two species Lilium longiflorum ,Oriental lily and floral organs of lily of four varieties - Diber, Siberia, Lilium Longiflorum and Lilium trompeten were selected for in vitro culture .Four stages of primary research were processed, that was, establishment of clone system, multiplication of embryonal callus, rooting and bulblet formation and transplantation. The result are shown as follows:
1. Stage of establishment of clone system
The bulblets of two species , Lilium Longiflorum and Oriental lily, were selected as explants for in vitro culture , and compared their contamination rate and inducing differentiation rate of different position in bulblet. The results of experiments indicated that the containminion rate of inner and middle part in bulblet were in low level. When different part in a single scale were cultured in the same medium the results showed that the basal part turned out to be the easiest part for inducing, the second was the middle part, the upper part was hard to induce. Among the inducing mediums in our experiment, MS medium supplemented with 2.0mg L-1 BA and 0.2 mg L-1 NAA was the best for Lilium Longiflorum , its inducing rate was 83 percent; while MS medium supplemented with 4.0mg L-1 BA and 0.2 mg L-1 NAA was suitable for Oriental lily, the inducing rate was 75 percent.
The floral organs of four varieties, Diber, Siberia, Lilium Longiflorum , and Lilium trompeten, were selected as explants for in vitro culture. It was founded that when they were culture in the same medium ,the inducing differentiation rate varied from varieties. Diber was the easiest variety for inducing which was followed by Siberia, Lilium Longiflorum, and Lilium trompeten. The inducing rates of different part in the same variety were also different. Their sequence, from easy to hard, was filament, floral receptable, ovary, corolla, style and anther. The explants were cultured in a series of inducing medium .The results showed that MS medium supplemented with 0.5mg L-1 BA and 0.5mg L-1 NAA and
MS medium supplemented with l.Omg'L"1 BA and O.Smg-L"1 NAA showed a better induction character.
When different part of bacteria and germ -free leafs were induced for embryonal callus, the basal part was the easiest to induce. MS medium supplemented with 2.0 mg.L"1 BA and 0.2mg.L"' NAA and MS medium contained l.Omg.L"1 BA and O.Zmg.L"1 NAA were suitable for basal part of leaf inducing culture.
2. Stage of multiplication and differentiation of embryonal callus
During the stage of embryonal callus of multiplication and differentiation ,it was found that, for Lilium Longiflorum bulblet and germ-free seeding , MS medium containing 0.2 mg.L"1 BA and 0.2 mg.L"1 NAA was the best medium; for Oriental lily, the best multiplication medium was MS supplemented with 1.0 mg.L"1 BA, and the best differentiation medium was MS supplemented with O.Smg.L"1 BA and 0.05 mg.L ' NAA; MS supplemented with l.Omg.L"1 BA and 0.2 mg.L"1 NAA were suitable for floral organs of Siberia and Diber ; and MS containing l.Omg.L"1 BAand O.Smg.L"1 NAA were fit to floral organs of Lilium Longiflorum .
3. Stage of rooting and bulblet formation
During the stage of rooting and bulblet formation, the result showed that the category and the concentration of hormone, the concentration of sugar and the concentration of macroelement, affected rooting and forming of bulblets. The application of PP333 benefited the bulblet formiation, the best concentration were 2.0 mg.L"1 and 2.5 mg.L"1. NAA promoted rooting but made against forming bulblet. Sucrose can help to form bulblet, the best concentration was 60g.L"1. It was favorable for rooting and bulblet formation in reducing the concentration of macroelement .1/4MS proved the best concentration.
4.Stage of transplantation
There were four stages in transplantation , that was sand bed prepairing , domestication, pseudoplantation and transplant soil and basin. The survival rate
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