东方百合离体培养与试管鳞茎诱导的研究
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摘要
本论文报道了对东方百合进行离体培养的研究结果,包括鳞片外植体的选择与灭菌、鳞茎芽的诱导分化、鳞茎芽的继代培养和增殖、试管鳞茎的诱导、试管苗的生根与移栽等,以建立较完善的东方百合组织培养技术体系。主要结果如下:
1、东方百合外层、中层鳞片较适宜的灭菌处理为:10%NaClO 10min+0.1%HgCl_2 25min;内层鳞片较适宜的灭菌处理为:10%NaClO 10min+0.1%HgCl_2 10min。
2、东方百合适宜的鳞片诱导分化培养基为:MS+6-BA 1.0mg/L+2,4-D 0.5mg/L+NAA 0.2mg/L。不同品种鳞片的芽诱导分化能力不同:索邦鳞茎芽诱导率71.45%,平均生成鳞茎芽数3.95;元帅鳞茎芽诱导率66.06%,平均生成鳞茎芽数3.48;西伯利亚鳞茎芽诱导率56.67%,平均生成鳞茎芽数2.09。同一品种不同部位的鳞片诱导能力也有差异,内层低于外层和中层。
3、三种基本培养基对东方百合鳞茎芽的增殖效应:MS＞Miller＞White。在这三类培养基中,索邦品种芽增殖倍数依次为:4.69、1.83、1.23;西伯利亚品种芽增殖倍数依次为:3.70、1.67、0.83;元帅品种芽增殖倍数依次为3.86、1.85、1.11。
4、一定浓度范围内的6-BA、KT和NAA、IBA间的配组对东方百合鳞茎芽的增殖起促进作用。鳞茎组培芽增殖的适宜培养基为:MS+1.0-2.0mg/L 6-BA+1.0mg/L KT+0.2mg/L NAA,不同品种对最佳激素组合浓度略有不同。3个品种间的芽增殖结果比较,以索邦最优。
5、不同细胞分裂素种类对芽增殖和壮苗的效果不同。6-BA在芽增殖方面优于同浓度水平的KT;KT在壮苗方面优于同浓度水平的6-BA。
6、东方百合增殖培养的适宜代数是3～5代。芽增殖倍数比较:索邦＞元帅＞西伯利亚。
7、适当提高蔗糖浓度能促进东方百合三个品种试管结鳞茎。当蔗糖浓度为80g/L时,东方百合三个品种的结鳞茎率、鳞茎直径和鳞茎鲜重均达到最大值。索邦结鳞茎率为100%,鳞茎直径1.768cm,鳞茎鲜重8.55g;西伯利亚结鳞茎率为100%,鳞茎直径1.709cm,鳞茎鲜重8.01g;元帅结鳞茎率为100%,鳞茎直径1.744cm,鳞茎鲜重8.82g。但当蔗糖至高浓度(＞100g/L)则对试管结鳞茎不利,所形成的鳞茎变小。
8、培养基中附加香豆素对试管结鳞茎有利。东方百合三个品种索邦、西伯利亚和元帅的香豆素适宜浓度分别为:12mg/L、6mg/L和9mg/L。
9、多效唑(MET)的适宜浓度对东方百合三个品种试管结鳞茎有促进作用。索邦的MET适宜浓度为3mg/L;西伯利亚和元帅的MET适宜浓度为2mg/L。
10、基本培养基1/2MS比MS更利于东方百合的生根。三个品种的苗素质和根素质比较:索邦＞西伯利亚＞元帅。
11、生长激素NAA、IBA浓度均以0.9mg/L时,适宜东方百合三个品种的生根培养。两种激素比较,NAA的生根效应优于IBA。
12、生根粉ABT有良好的发根作用。当ABT浓度为4.0mg/L时,索邦根数最多,为17.95条;当ABT浓度为3.0mg/L时,元帅根数最多,为13.45条;当ABT浓度为2.0mg/L时,西伯利亚根数最多,为12.67条。
13、适宜浓度(0.6-0.9mg/L)的多效唑能使苗健壮,苗移栽后成活率也较高。MET浓度过高(＞1.2mg/L)会抑制试管苗的生长。
14、不同种类基质对试管苗移栽生长影响不同。单一基质比较:泥炭＞菜园土＞细沙＞黄泥,索邦品种在泥炭中的成活率为98.91%,西伯利亚为84.62%,元帅为90.00%。复合基质效果优于单一基质,泥炭1/2+珍珠岩1/4+菜园土1/4是东方百合三个品种较为理想栽培基质。在此复合基质中,索邦品种成活率为100%,西伯利亚为90.00%,元帅为95.90%。品种间移栽成活率、抗逆性、适应力比较:索邦＞元帅＞西伯利亚。
This paper reported the studied results on culture in vitro of Oriental lily varieties, which included selection and sterilization of squamas explants, induction and differentiation of bulb buds, subculture and propagation of bulb buds, induction and formation of test-tube bulblets, rooting and transplanting of test-tube plantlets, etc, in order to set up the better system on tissue culture of Oriental Lily. The main results indicated as follows:
1、The more suitable sterilization treatments of outer and middle squamas of Oriental lily was 10% NaClO 10min + 0.1% HgCl_225min and that of inner squamas of Oriental lily was 10% NaClO 10min + 0.1 % HgCl_2 10min.
2、The appropriate medium for the squamas induction and differentiation of Oriental Lily was MS + 6- BA 1.0mg/L + 2,4-D 0.5mg/L + NAA 0.2mg/L. There were different frequencies of buds induction and differentiation with differentiation species squamas: the bud induction rate of Sorbonne was 71.45%, and the average number of bulblets formation was 3.95; the bud induction rate of Acapulco was 66.06%, and the average number of bulblets formation was 3.48; the bud induction rate of Siberia was 56.67%, and the average number of bulblets formation was 2.09. There were also differences on the induction ability in different parts of the squamas in the same species: the inner was lower than the outer and the middle.
3、The effects on buds propagation of three basic media of Oriental Lily's bulbs was MS>Miller> White. In these three categories of media, Sorbonne's multiple of bud multiplication ordinally were 4.69, 1.83, 1.23; Siberia's multiple of bud multiplication ordinally were 3.70, 1.67, 0.83; Acapulco's multiple of bud multiplication ordinally were 3.86, 1.85, 1.11.
4、A certain concentration within the scope of 6-BA and KT combined with NAA or IBA play the facilitating role on proliferation of bulb culture buds. The optimal medium for buds propagation was: MS+1.0-2.0mg/L 6-BA+1.0mg/L KT+0.2mg/L NAA. The best combination of hormone concentration of three varieties differ slightly. Comparing with the buds propagation results between three varieties, Sorbonne was the best.
5、There were different effects on the buds propagation and sound plantlets in different kinds of cytokimin. At the same concentration level, 6-BA was better than KT in the buds propagation and KT was better than 6-BA in sound plantlet aspect.
6、The appropriate generations for buds propagation of Oriental Lily was generations three to five. The comparison of bud propagation multiples to three varieties was Sorbonne >Acapulco> Siberia.
7、Increasing appropriately the concentration of sugar could promote the bulblets formation to three varieties of Oriental Lily. When sucrose concentration was 80g/L, the rate of bublet formation and the diameter and the weight of bulblet were achived to the biggest value. Under this sucrose concentration, the bulblet formation rate of Sorbonne reached 100%, the diameter of bulblet was 1.768cm, and the weight of blublet was 8.55g; the bulblet formation rate of Siberia reached 100%, the diameter of bulblet was 1.709cm, and the weight of blublet was 8.01g; the bulblet formation rate of Acapulco reached 100%, the diameter of bulbler was 1.744cm, and the weight of blublet was 8.82g. But the high sucrose concentration (>100g/L) was detrimental to the test-tube bulb and the bulb formed smaller.
8、Added cumarin to the medium could promot bulblet formation.The concentrations of cumarin for the three varieties of Oriental Lily (Sorbonne, Siberia and Acapulco) were 12mg/L, 6mg/L and 9mg/L.
9、The appropriate concentration of MET could promot bulblet formation of the three varieties of Oriental Lily. MET 3mg/L was availbe to Sorbonne and MET 2mg/L was suitable to Siberia and Acapulco.
10、Basic medium 1/2 MS more suitble than MS to the rooting of Oriental Lily. The comparison of plantlet quality and root quality to the three varieties was Sorbonne > Siberia > Acapulco.
11、When growth hormone NAA, IBA concentration was 0.9mg/L, the concentration was suitble to rooting culture of Oriental Lily. Comparing the two hormones, NAA was superior to IBA on rooting effect.
12、ABT played a good role in rooting. When ABT concentration was 4.0mg/L, the most root number of Sorbonne was 17.95; when its concentration was 3.0mg/L, the most root number of Acapulco was 13.45; when its concentration was 2.0 mg/L, the most root number of Siberian was 12.67.
13、The appropriate concentration of MET (0.6-0.9mg/L) could make the plantlets stronger and the survival rate after transplanting was higher. High concentrations (> 1.2mg/L) of MET could inhibit the growth of the plantlets.
14、There were different effects of different kinds of matrix on the transplantation and growth of plantlets. The comparison of a single matrix was peat >garden soil > sand > mud. The survival rate of Sorbonne in the peat was 98.91%, when Siberia and Acapulco showed 84.62% and 90.00%. Multi-matrix was better than a single matrix. Peat 1 / 2 + perlite 1 / 4 + garden soil 1/4 was the better matrix for the three varieties of Oriental Lily. In this multi-matrix, the survival rate of Sorbonne was 100%, Siberia and Acapulco showed 90.00% and 95.90%. Comparing with the transplant survival rate, the resistance and the resilience between the varieties, the order was Sorbonne > Acapulco > Siberia.
1、东方百合外层、中层鳞片较适宜的灭菌处理为:10%NaClO 10min+0.1%HgCl_2 25min;内层鳞片较适宜的灭菌处理为:10%NaClO 10min+0.1%HgCl_2 10min。
2、东方百合适宜的鳞片诱导分化培养基为:MS+6-BA 1.0mg/L+2,4-D 0.5mg/L+NAA 0.2mg/L。不同品种鳞片的芽诱导分化能力不同:索邦鳞茎芽诱导率71.45%,平均生成鳞茎芽数3.95;元帅鳞茎芽诱导率66.06%,平均生成鳞茎芽数3.48;西伯利亚鳞茎芽诱导率56.67%,平均生成鳞茎芽数2.09。同一品种不同部位的鳞片诱导能力也有差异,内层低于外层和中层。
3、三种基本培养基对东方百合鳞茎芽的增殖效应:MS＞Miller＞White。在这三类培养基中,索邦品种芽增殖倍数依次为:4.69、1.83、1.23;西伯利亚品种芽增殖倍数依次为:3.70、1.67、0.83;元帅品种芽增殖倍数依次为3.86、1.85、1.11。
4、一定浓度范围内的6-BA、KT和NAA、IBA间的配组对东方百合鳞茎芽的增殖起促进作用。鳞茎组培芽增殖的适宜培养基为:MS+1.0-2.0mg/L 6-BA+1.0mg/L KT+0.2mg/L NAA,不同品种对最佳激素组合浓度略有不同。3个品种间的芽增殖结果比较,以索邦最优。
5、不同细胞分裂素种类对芽增殖和壮苗的效果不同。6-BA在芽增殖方面优于同浓度水平的KT;KT在壮苗方面优于同浓度水平的6-BA。
6、东方百合增殖培养的适宜代数是3～5代。芽增殖倍数比较:索邦＞元帅＞西伯利亚。
7、适当提高蔗糖浓度能促进东方百合三个品种试管结鳞茎。当蔗糖浓度为80g/L时,东方百合三个品种的结鳞茎率、鳞茎直径和鳞茎鲜重均达到最大值。索邦结鳞茎率为100%,鳞茎直径1.768cm,鳞茎鲜重8.55g;西伯利亚结鳞茎率为100%,鳞茎直径1.709cm,鳞茎鲜重8.01g;元帅结鳞茎率为100%,鳞茎直径1.744cm,鳞茎鲜重8.82g。但当蔗糖至高浓度(＞100g/L)则对试管结鳞茎不利,所形成的鳞茎变小。
8、培养基中附加香豆素对试管结鳞茎有利。东方百合三个品种索邦、西伯利亚和元帅的香豆素适宜浓度分别为:12mg/L、6mg/L和9mg/L。
9、多效唑(MET)的适宜浓度对东方百合三个品种试管结鳞茎有促进作用。索邦的MET适宜浓度为3mg/L;西伯利亚和元帅的MET适宜浓度为2mg/L。
10、基本培养基1/2MS比MS更利于东方百合的生根。三个品种的苗素质和根素质比较:索邦＞西伯利亚＞元帅。
11、生长激素NAA、IBA浓度均以0.9mg/L时,适宜东方百合三个品种的生根培养。两种激素比较,NAA的生根效应优于IBA。
12、生根粉ABT有良好的发根作用。当ABT浓度为4.0mg/L时,索邦根数最多,为17.95条;当ABT浓度为3.0mg/L时,元帅根数最多,为13.45条;当ABT浓度为2.0mg/L时,西伯利亚根数最多,为12.67条。
13、适宜浓度(0.6-0.9mg/L)的多效唑能使苗健壮,苗移栽后成活率也较高。MET浓度过高(＞1.2mg/L)会抑制试管苗的生长。
14、不同种类基质对试管苗移栽生长影响不同。单一基质比较:泥炭＞菜园土＞细沙＞黄泥,索邦品种在泥炭中的成活率为98.91%,西伯利亚为84.62%,元帅为90.00%。复合基质效果优于单一基质,泥炭1/2+珍珠岩1/4+菜园土1/4是东方百合三个品种较为理想栽培基质。在此复合基质中,索邦品种成活率为100%,西伯利亚为90.00%,元帅为95.90%。品种间移栽成活率、抗逆性、适应力比较:索邦＞元帅＞西伯利亚。
This paper reported the studied results on culture in vitro of Oriental lily varieties, which included selection and sterilization of squamas explants, induction and differentiation of bulb buds, subculture and propagation of bulb buds, induction and formation of test-tube bulblets, rooting and transplanting of test-tube plantlets, etc, in order to set up the better system on tissue culture of Oriental Lily. The main results indicated as follows:
1、The more suitable sterilization treatments of outer and middle squamas of Oriental lily was 10% NaClO 10min + 0.1% HgCl_225min and that of inner squamas of Oriental lily was 10% NaClO 10min + 0.1 % HgCl_2 10min.
2、The appropriate medium for the squamas induction and differentiation of Oriental Lily was MS + 6- BA 1.0mg/L + 2,4-D 0.5mg/L + NAA 0.2mg/L. There were different frequencies of buds induction and differentiation with differentiation species squamas: the bud induction rate of Sorbonne was 71.45%, and the average number of bulblets formation was 3.95; the bud induction rate of Acapulco was 66.06%, and the average number of bulblets formation was 3.48; the bud induction rate of Siberia was 56.67%, and the average number of bulblets formation was 2.09. There were also differences on the induction ability in different parts of the squamas in the same species: the inner was lower than the outer and the middle.
3、The effects on buds propagation of three basic media of Oriental Lily's bulbs was MS>Miller> White. In these three categories of media, Sorbonne's multiple of bud multiplication ordinally were 4.69, 1.83, 1.23; Siberia's multiple of bud multiplication ordinally were 3.70, 1.67, 0.83; Acapulco's multiple of bud multiplication ordinally were 3.86, 1.85, 1.11.
4、A certain concentration within the scope of 6-BA and KT combined with NAA or IBA play the facilitating role on proliferation of bulb culture buds. The optimal medium for buds propagation was: MS+1.0-2.0mg/L 6-BA+1.0mg/L KT+0.2mg/L NAA. The best combination of hormone concentration of three varieties differ slightly. Comparing with the buds propagation results between three varieties, Sorbonne was the best.
5、There were different effects on the buds propagation and sound plantlets in different kinds of cytokimin. At the same concentration level, 6-BA was better than KT in the buds propagation and KT was better than 6-BA in sound plantlet aspect.
6、The appropriate generations for buds propagation of Oriental Lily was generations three to five. The comparison of bud propagation multiples to three varieties was Sorbonne >Acapulco> Siberia.
7、Increasing appropriately the concentration of sugar could promote the bulblets formation to three varieties of Oriental Lily. When sucrose concentration was 80g/L, the rate of bublet formation and the diameter and the weight of bulblet were achived to the biggest value. Under this sucrose concentration, the bulblet formation rate of Sorbonne reached 100%, the diameter of bulblet was 1.768cm, and the weight of blublet was 8.55g; the bulblet formation rate of Siberia reached 100%, the diameter of bulblet was 1.709cm, and the weight of blublet was 8.01g; the bulblet formation rate of Acapulco reached 100%, the diameter of bulbler was 1.744cm, and the weight of blublet was 8.82g. But the high sucrose concentration (>100g/L) was detrimental to the test-tube bulb and the bulb formed smaller.
8、Added cumarin to the medium could promot bulblet formation.The concentrations of cumarin for the three varieties of Oriental Lily (Sorbonne, Siberia and Acapulco) were 12mg/L, 6mg/L and 9mg/L.
9、The appropriate concentration of MET could promot bulblet formation of the three varieties of Oriental Lily. MET 3mg/L was availbe to Sorbonne and MET 2mg/L was suitable to Siberia and Acapulco.
10、Basic medium 1/2 MS more suitble than MS to the rooting of Oriental Lily. The comparison of plantlet quality and root quality to the three varieties was Sorbonne > Siberia > Acapulco.
11、When growth hormone NAA, IBA concentration was 0.9mg/L, the concentration was suitble to rooting culture of Oriental Lily. Comparing the two hormones, NAA was superior to IBA on rooting effect.
12、ABT played a good role in rooting. When ABT concentration was 4.0mg/L, the most root number of Sorbonne was 17.95; when its concentration was 3.0mg/L, the most root number of Acapulco was 13.45; when its concentration was 2.0 mg/L, the most root number of Siberian was 12.67.
13、The appropriate concentration of MET (0.6-0.9mg/L) could make the plantlets stronger and the survival rate after transplanting was higher. High concentrations (> 1.2mg/L) of MET could inhibit the growth of the plantlets.
14、There were different effects of different kinds of matrix on the transplantation and growth of plantlets. The comparison of a single matrix was peat >garden soil > sand > mud. The survival rate of Sorbonne in the peat was 98.91%, when Siberia and Acapulco showed 84.62% and 90.00%. Multi-matrix was better than a single matrix. Peat 1 / 2 + perlite 1 / 4 + garden soil 1/4 was the better matrix for the three varieties of Oriental Lily. In this multi-matrix, the survival rate of Sorbonne was 100%, Siberia and Acapulco showed 90.00% and 95.90%. Comparing with the transplant survival rate, the resistance and the resilience between the varieties, the order was Sorbonne > Acapulco > Siberia.
引文
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