凤丹体胚发生及愈伤组织诱导芽分化研究
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摘要
本文研究了牡丹不同时期的种胚直接诱导体细胞胚,并在此基础上实现了体细胞胚植株再生。利用牡丹的叶片、茎段、花药、花瓣进行了愈伤组织诱导,获得愈伤组织的基础上进行增殖和分化培养获得了小植株,并实现了驯化移栽。同时利用石蜡切片和扫描电镜的方法对愈伤组织发育过程中的结构变化进行了观察。主要研究结果如下：
在牡丹体细胞胚诱导方面,本试验结果表明利用‘凤丹’花后110d的种胚诱导率最高。种胚、子叶、胚轴3种外植体进行体细胞胚的诱导,种胚的诱导率最高,为32.5%。利用90g/L的蔗糖溶液处理种胚2h,体细胞胚诱导率为38.0%。对花后110d的种胚进行体细胞胚诱导,‘凤丹’体细胞胚增殖最适合的培养基组合为：MS+6-BA0.1mg/L+蔗糖60.0 g/L+CH0.4 g/L,体细胞胚诱导率为38.33%。
体细胞胚成熟过程中,活性炭对牡丹体细胞胚成熟具有显著的促进作用,而不同的培养基对体细胞胚成熟效果差异不显著。6-BA能够促进体细胞胚的萌发,GA3对打破休眠促进萌发具有较好的效果。IAA对促进体细胞胚生根效果较好。本文首次探讨‘凤丹’体细胞胚直接诱导,首次深入研究了添加物质CH和活性炭对体细胞胚诱导及成熟的影响,首次对‘凤丹’进行体细胞胚诱导,诱导率均在15%以上,最高诱导率达到38.33%,并成功获得了体细胞胚植株。
牡丹叶片愈伤组织诱导过程中,改良MS培养基为诱导牡丹叶片愈伤组织的最佳培养基,在未添加任何植物植物激素的培养基上无法诱导出愈伤组织,单独添加生长素或细胞分裂素均能诱导出愈伤组织,但诱导率有差异,最佳的生长素是2,4-D 0.2 mg/L,最佳的细胞分裂素是TDZ 2.0 mg/L。用2,4-D、NAA、TDZ正交试验得到最佳组合为改良MS+2,4-D 0.2 mg/L+NAA 0.2 mg/L+TDZ 3.0 mg/L,愈伤诱导率高达87.8%。利用牡丹幼嫩茎段为外植体,进行愈伤组织诱导,通过试验得出最佳的培养基组合为MS+NAA0.05mg/L+6-BA2.0 mg/L+2,4-D1.0 mg/L为最佳的培养基组合,在此培养基诱导下,茎段愈伤组织诱导率达到最高的87.8%。牡丹茎段愈伤组织诱导率在不同蔗糖浓度时的大小顺序为30.0 g/L>40.0 g/L>20.0 g/L>10.0 g/L,因此最佳的蔗糖浓度为30.0 g/L。利用茎段的中下部、将外植体横放在培养基上会获得较高的愈伤组织诱导率和较多的愈伤组织。试验证明,牡丹花药诱导愈伤组织的最佳时期为花粉的单核中期,最佳的培养基为MS+2,4-D2.0 mg/L+6-BA1.5 mg/L+NAA1.0 mg/L,在最佳培养基、蔗糖质量浓度为6%,4℃的低温条件下处理8d的愈伤组织诱导率最高达到50.8%。利用花瓣诱导愈伤组织,最佳的培养基组合为MS+2,4-D 2.0 mg/L+6-BA 1.5 mg/L+NAA 0.3 mg/L,诱导率达到98.9%。
牡丹愈伤组织增殖过程中,叶片愈伤组织最合适的增殖培养基为MS+NAA0.2mg/L+6-BA2.0+KT2.0 mg/L,增殖率为209.8%。不同碳源对愈伤组织增殖的影响不同,30.0g/L的蔗糖为合适碳源。添加浓度为0.4g/L的CH效果最好。试验结果表明叶片愈伤组织增殖的生长曲线为S型曲线,生长分为延缓生长期、指数生长期和静止期三个时期,生长周期为30d左右。茎段和花瓣的增殖培养基相同,均为MS+NAA0.2mg/L+6-BA3.0mg/L+蔗糖30.0g/L+CH0.4g/L+琼脂7.0g/L,增殖率分别为237.3%和233.5%。而花药的增殖培养基为MS+NAA0.2mg/L+6-BA2.0mg/L+蔗糖30.0g/L+CH0.4g/L+琼脂7.0g/L,增殖率为230.9%。光照12h对牡丹茎段愈伤组织的增殖效果最好。
研究了光照强度、抗氧化剂和吸附剂对愈伤组织褐化率的影响,认为在转接初期将所有愈伤组织,放置在黑暗条件下培养1周左右后转移到光照强度为19umol/m2/s下培养,既能够保证愈伤组织的增殖生长,又降低愈伤组织在增殖过程中的褐化。几种抗氧化剂能够大大降低愈伤组织褐化率。其中AgNO3对愈伤组织褐化的抑制效果最好。对于各种吸附剂而言,1.0g/LPVP和2.0g/L活性炭两种吸附剂抑制褐化效果最好。
牡丹愈伤组织结构细胞学观察。牡丹的芽原基不是从愈伤组织表面发生,而是发生于愈伤组织的近表层,然后突破表层细胞开始发育。通过石蜡切片发现在不同的发育阶段不同形状的胚状体同时存在,不同的愈伤组织细胞结构差异较大。
通过扫描电镜可以看出,不同的愈伤组织培养的不同时间会发生不同的变化,有的愈伤组织可以发育成胚状体,有的则无法完成正常的发育,同一块愈伤组织可能存在胚性愈伤组织和非胚性愈伤组织两种不同的类型。从牡丹愈伤组织的发育过程中来看,继代后15-20d是愈伤组织发育最快的时期,经过这一时期后愈伤组织发育开始减慢。
在牡丹愈伤组织分化过程中,最合适的叶片愈伤组织分化培养基为MS+6-BA2.0mg/L+NAA0.2mg/L+TDZ0.3 mg/L+蔗糖30.0g/L+琼脂7.0g/L,分化率为22.22%。茎段愈伤组织最适合的分化培养基为MS+6-BA3.0 mg/L+NAA0.2mg/L,分化率为24.44%。花药愈伤组织分化的最佳培养基为MS+6-BA2.0 mg/L+NAA0.2 mg/L+KT0.3mg/L,分化率为22.22%,干燥处理3d有利于花药愈伤组织的分化。花瓣愈伤组织分化的合适培养基为MS+ZT 0.5 mg/L+6-BA2.0 mg/L,分化率为34.81%。
对于愈伤组织诱导形成的植株,在各种不同培养基上诱导生根,1/2MS和WPM两种培养基是合适的培养基,最适合的蔗糖浓度为30.0g/L。最佳的培养基组合为1/2MS+NAA0.1mg/L+IBA0.2mg/L+蔗糖30.0g/L。小植株在驯化后能够移栽成活,成活率为40.0%左右。
In this paper, Different stages of Peony embryo were used to directly induce somatic embryos. peony leaves, stems, anthers, petals were used to induce the callus,then through callus proliferation and differentiation to achieve small plants. Based on the induction of small plant, The roots were induced in this experiment. At last acclimation and transplanting were succeed. At the same time using paraffin sections and scanning electron microscopy methods observed the callus structural changes during development.The main results were as follows:
Somatic embryo induction of peony. The seed embryo of peony after flower 110d of 'Feng Dan'were used to induce the somatic embryo,the induction rate was higher than other stages. Embryo, cotyledon, hypocotyl explants were all able to induce somatic embryos,but the highest induction rate were appeared while using embryo as explants. Sucrose pretreatment to embryo were helpful to somatic embryos induction.The experiment showed that using 90g/L sucrose dealing with embryo 2h were best. The best medium of somatic embryos induction were MS+2,4-D 0.2mg/L+6-BA 0.2mg/L.With this medium, the rate of embryo induction was highest of all. Peony proliferation of somatic embryos and plant regeneration. The results showed that the most suitable culture medium of peony'Feng Dan'the proliferation were MS+6-BA0.1 mg/L+sucrose 60.0 g/L+CH0.4 g/L. Activated charcoal played significant role in somatic embryo maturation and different culture media on somatic embryo maturation effect was not significant.6-BA can promote the germination of somatic embryos, but the excess of it will make somatic embryos more callus and thus can not be achieved plant regeneration. GA3 were good for break dormancy and promote germination. IAA were good at promoting rooting of somatic embryos.
Callus induction of peony. The best medium for callus induction of peony leaves was Modified MS medium.Without adding any plant growth regulator medium can not induce callus. Adding auxin or cytokinin alone can induce callus organization, but there were differences in induction rates. The best auxin was 2,4-D 0.2 mg/L and the best cytokinin was TDZ 2.0 mg/L. Using 2,4-D, NAA, TDZ for orthogonal experiment, the optimal medium were modified MS+2,4-D 0.2 mg/L+NAA 0.2 mg/L+TDZ 3.0 mg/L and the induction rate of callus was 87.8%.The tender stems of peony were used as explants for callus induction,The best medium were MS+NAA0.05 mg/L+6-BA2.0 mg/L+2,4-D1.0 mg/L.In this medium, the highest induction rate of callus was 87.8%. The order of sucrose concentrations to peony stem callus induction were 30.0 g/L> 40.0 g/L> 20.0g/L> 10.0 g/L, so the best sucrose concentration was 30.0 g/L. Using the lower and middle stem were better than other oposition.With this stems, callus induction rate will be higher and callus will be more.The results showed that the best period was the middle develop stage. The best growth regulators were MS+2,4-D2.0 mg/L+6-BA1.5 mg/L+NAA1.0 mg/L. The best sucrose concentration was 6%, After 8 days cold pre-treatment in 4℃, the callus grew well, the induction efficiency reached to 50.8%.The petals were used as explants in this experiment, The best medium wereMS+2,4-D 2.0 mg/L+6-BA 1.5 mg/L+NAA 0.3 mg/L,The induction rate reached to 98.9%.
In this experiment,The roots of peony were used as explants,but there were no callus appeared, the reason of it probably due to the root fungi are more difficult to sterilization, It was likely to cause pollution. After inoculated root to the medium for a period of time, The root got brown black and there were no callus appeared.
The most appropriate medium for callus proliferation of leaves were MS+NAA0.2 mg/L +6-BA2.0 mg/L+KT2.0 mg/L.30.0 g/L of sucrose as a suitable carbon source for callus proliferation. CH with 0.4 g/L was good for callus proliferation. The results showed that the growth curve of leaf callus was the S-curve, The grow process can be divided into slow growth phase, exponential phase and stationary phase, The growth period of leaf callus was about 30d.The callus proliferation medium of Stems and petals were same, They were MS+ NAA0.2 mg/L+6-BA3.0 mg/L+sucrose 30.0 g/L+CH0.4 g/L+Agar 7.0 g/L. The medium of anther proliferation were MS+NAA0.2 mg/L+6-BA2.0 mg/L+sucrose 30.0 g/L+CH0.4 g/L+Agar 7.0 g/L.12h light was good for stem proliferation.
The effects of light intensity, anti-oxidants and adsorbents to callus browning were studied in this experiment. The results showed that after dark conditions for 1 week and then cultured in light intensity 19umol/m2/s may be good choice, In this condition,not only ensure the callus proliferation but also greatly reduce the callus browning. Several antioxidants had good effect to callus proliferation and reduced the rate of callus browning. AgNO3 was good for inhibiting the browning. 1.0g/LPVP and 2.0g/L activated carbon were good for inhibiting browning.
Callus structure observation of peony. The result showed that bud primordia are not from the surface of callus but occurred near the surface, and then began to break through the surface cell to grow. Different developmental stages were found in different shapes.
Using scanning electron microscopy, different callus culture occurs at different times different changes can be found, some callus can develop into embryoids, while others are unable to complete normal development. embryogenic callus and non-embryogenic callus may exist on one callus.15-20d after subculture ofth was the fastest time, after this period callus began to grow slow,so after the 15-20d of subculture is more good subculture period.
Callus differentiation of peony. The most suitable medium for callus differentiation of leaves were MS+6-BA2.0 mg/L+NAA0.2 mg/L+TDZ0.3 mg/L+sucrose 30.0 g/L+Agar 7.0 g/L. CH had little effect on the callus differentiation., The highest callus differentiation rate achieved when the sucrose concentration of 30g/L. Activated carbon were negative to callus differentiation, but good for the growth of adventitious buds.
The most appropriate callus differentiation medium of stem were MS+6-BA3.0 mg/L+ NAA0.2 mg/L. That of anther were MS+6-BA2.0 mg/L+NAA0.2 mg/L+KT0.3 mg/L, After dried 3d with filter paper,the anther callus formation quickly. The suitable medium of petal callus differentiation were MS+ZT 0.5 mg/L+6-BA2.0 mg/L.
Root induction and transplanting of peony. Four mediums were used in rooting induction,The results showed that 1/2MSand WPM were appropriate medium.The suitable sucrose concentration was 30.0g/L. The best medium were 1/2MS+NAA0.1mg/L+ IBA0.2mg/L+sucrose 30.0 g/L. Small plants can be transplanted after the domestication of survival, the survival rate was 40.0% or so.
在牡丹体细胞胚诱导方面,本试验结果表明利用‘凤丹’花后110d的种胚诱导率最高。种胚、子叶、胚轴3种外植体进行体细胞胚的诱导,种胚的诱导率最高,为32.5%。利用90g/L的蔗糖溶液处理种胚2h,体细胞胚诱导率为38.0%。对花后110d的种胚进行体细胞胚诱导,‘凤丹’体细胞胚增殖最适合的培养基组合为：MS+6-BA0.1mg/L+蔗糖60.0 g/L+CH0.4 g/L,体细胞胚诱导率为38.33%。
体细胞胚成熟过程中,活性炭对牡丹体细胞胚成熟具有显著的促进作用,而不同的培养基对体细胞胚成熟效果差异不显著。6-BA能够促进体细胞胚的萌发,GA3对打破休眠促进萌发具有较好的效果。IAA对促进体细胞胚生根效果较好。本文首次探讨‘凤丹’体细胞胚直接诱导,首次深入研究了添加物质CH和活性炭对体细胞胚诱导及成熟的影响,首次对‘凤丹’进行体细胞胚诱导,诱导率均在15%以上,最高诱导率达到38.33%,并成功获得了体细胞胚植株。
牡丹叶片愈伤组织诱导过程中,改良MS培养基为诱导牡丹叶片愈伤组织的最佳培养基,在未添加任何植物植物激素的培养基上无法诱导出愈伤组织,单独添加生长素或细胞分裂素均能诱导出愈伤组织,但诱导率有差异,最佳的生长素是2,4-D 0.2 mg/L,最佳的细胞分裂素是TDZ 2.0 mg/L。用2,4-D、NAA、TDZ正交试验得到最佳组合为改良MS+2,4-D 0.2 mg/L+NAA 0.2 mg/L+TDZ 3.0 mg/L,愈伤诱导率高达87.8%。利用牡丹幼嫩茎段为外植体,进行愈伤组织诱导,通过试验得出最佳的培养基组合为MS+NAA0.05mg/L+6-BA2.0 mg/L+2,4-D1.0 mg/L为最佳的培养基组合,在此培养基诱导下,茎段愈伤组织诱导率达到最高的87.8%。牡丹茎段愈伤组织诱导率在不同蔗糖浓度时的大小顺序为30.0 g/L>40.0 g/L>20.0 g/L>10.0 g/L,因此最佳的蔗糖浓度为30.0 g/L。利用茎段的中下部、将外植体横放在培养基上会获得较高的愈伤组织诱导率和较多的愈伤组织。试验证明,牡丹花药诱导愈伤组织的最佳时期为花粉的单核中期,最佳的培养基为MS+2,4-D2.0 mg/L+6-BA1.5 mg/L+NAA1.0 mg/L,在最佳培养基、蔗糖质量浓度为6%,4℃的低温条件下处理8d的愈伤组织诱导率最高达到50.8%。利用花瓣诱导愈伤组织,最佳的培养基组合为MS+2,4-D 2.0 mg/L+6-BA 1.5 mg/L+NAA 0.3 mg/L,诱导率达到98.9%。
牡丹愈伤组织增殖过程中,叶片愈伤组织最合适的增殖培养基为MS+NAA0.2mg/L+6-BA2.0+KT2.0 mg/L,增殖率为209.8%。不同碳源对愈伤组织增殖的影响不同,30.0g/L的蔗糖为合适碳源。添加浓度为0.4g/L的CH效果最好。试验结果表明叶片愈伤组织增殖的生长曲线为S型曲线,生长分为延缓生长期、指数生长期和静止期三个时期,生长周期为30d左右。茎段和花瓣的增殖培养基相同,均为MS+NAA0.2mg/L+6-BA3.0mg/L+蔗糖30.0g/L+CH0.4g/L+琼脂7.0g/L,增殖率分别为237.3%和233.5%。而花药的增殖培养基为MS+NAA0.2mg/L+6-BA2.0mg/L+蔗糖30.0g/L+CH0.4g/L+琼脂7.0g/L,增殖率为230.9%。光照12h对牡丹茎段愈伤组织的增殖效果最好。
研究了光照强度、抗氧化剂和吸附剂对愈伤组织褐化率的影响,认为在转接初期将所有愈伤组织,放置在黑暗条件下培养1周左右后转移到光照强度为19umol/m2/s下培养,既能够保证愈伤组织的增殖生长,又降低愈伤组织在增殖过程中的褐化。几种抗氧化剂能够大大降低愈伤组织褐化率。其中AgNO3对愈伤组织褐化的抑制效果最好。对于各种吸附剂而言,1.0g/LPVP和2.0g/L活性炭两种吸附剂抑制褐化效果最好。
牡丹愈伤组织结构细胞学观察。牡丹的芽原基不是从愈伤组织表面发生,而是发生于愈伤组织的近表层,然后突破表层细胞开始发育。通过石蜡切片发现在不同的发育阶段不同形状的胚状体同时存在,不同的愈伤组织细胞结构差异较大。
通过扫描电镜可以看出,不同的愈伤组织培养的不同时间会发生不同的变化,有的愈伤组织可以发育成胚状体,有的则无法完成正常的发育,同一块愈伤组织可能存在胚性愈伤组织和非胚性愈伤组织两种不同的类型。从牡丹愈伤组织的发育过程中来看,继代后15-20d是愈伤组织发育最快的时期,经过这一时期后愈伤组织发育开始减慢。
在牡丹愈伤组织分化过程中,最合适的叶片愈伤组织分化培养基为MS+6-BA2.0mg/L+NAA0.2mg/L+TDZ0.3 mg/L+蔗糖30.0g/L+琼脂7.0g/L,分化率为22.22%。茎段愈伤组织最适合的分化培养基为MS+6-BA3.0 mg/L+NAA0.2mg/L,分化率为24.44%。花药愈伤组织分化的最佳培养基为MS+6-BA2.0 mg/L+NAA0.2 mg/L+KT0.3mg/L,分化率为22.22%,干燥处理3d有利于花药愈伤组织的分化。花瓣愈伤组织分化的合适培养基为MS+ZT 0.5 mg/L+6-BA2.0 mg/L,分化率为34.81%。
对于愈伤组织诱导形成的植株,在各种不同培养基上诱导生根,1/2MS和WPM两种培养基是合适的培养基,最适合的蔗糖浓度为30.0g/L。最佳的培养基组合为1/2MS+NAA0.1mg/L+IBA0.2mg/L+蔗糖30.0g/L。小植株在驯化后能够移栽成活,成活率为40.0%左右。
In this paper, Different stages of Peony embryo were used to directly induce somatic embryos. peony leaves, stems, anthers, petals were used to induce the callus,then through callus proliferation and differentiation to achieve small plants. Based on the induction of small plant, The roots were induced in this experiment. At last acclimation and transplanting were succeed. At the same time using paraffin sections and scanning electron microscopy methods observed the callus structural changes during development.The main results were as follows:
Somatic embryo induction of peony. The seed embryo of peony after flower 110d of 'Feng Dan'were used to induce the somatic embryo,the induction rate was higher than other stages. Embryo, cotyledon, hypocotyl explants were all able to induce somatic embryos,but the highest induction rate were appeared while using embryo as explants. Sucrose pretreatment to embryo were helpful to somatic embryos induction.The experiment showed that using 90g/L sucrose dealing with embryo 2h were best. The best medium of somatic embryos induction were MS+2,4-D 0.2mg/L+6-BA 0.2mg/L.With this medium, the rate of embryo induction was highest of all. Peony proliferation of somatic embryos and plant regeneration. The results showed that the most suitable culture medium of peony'Feng Dan'the proliferation were MS+6-BA0.1 mg/L+sucrose 60.0 g/L+CH0.4 g/L. Activated charcoal played significant role in somatic embryo maturation and different culture media on somatic embryo maturation effect was not significant.6-BA can promote the germination of somatic embryos, but the excess of it will make somatic embryos more callus and thus can not be achieved plant regeneration. GA3 were good for break dormancy and promote germination. IAA were good at promoting rooting of somatic embryos.
Callus induction of peony. The best medium for callus induction of peony leaves was Modified MS medium.Without adding any plant growth regulator medium can not induce callus. Adding auxin or cytokinin alone can induce callus organization, but there were differences in induction rates. The best auxin was 2,4-D 0.2 mg/L and the best cytokinin was TDZ 2.0 mg/L. Using 2,4-D, NAA, TDZ for orthogonal experiment, the optimal medium were modified MS+2,4-D 0.2 mg/L+NAA 0.2 mg/L+TDZ 3.0 mg/L and the induction rate of callus was 87.8%.The tender stems of peony were used as explants for callus induction,The best medium were MS+NAA0.05 mg/L+6-BA2.0 mg/L+2,4-D1.0 mg/L.In this medium, the highest induction rate of callus was 87.8%. The order of sucrose concentrations to peony stem callus induction were 30.0 g/L> 40.0 g/L> 20.0g/L> 10.0 g/L, so the best sucrose concentration was 30.0 g/L. Using the lower and middle stem were better than other oposition.With this stems, callus induction rate will be higher and callus will be more.The results showed that the best period was the middle develop stage. The best growth regulators were MS+2,4-D2.0 mg/L+6-BA1.5 mg/L+NAA1.0 mg/L. The best sucrose concentration was 6%, After 8 days cold pre-treatment in 4℃, the callus grew well, the induction efficiency reached to 50.8%.The petals were used as explants in this experiment, The best medium wereMS+2,4-D 2.0 mg/L+6-BA 1.5 mg/L+NAA 0.3 mg/L,The induction rate reached to 98.9%.
In this experiment,The roots of peony were used as explants,but there were no callus appeared, the reason of it probably due to the root fungi are more difficult to sterilization, It was likely to cause pollution. After inoculated root to the medium for a period of time, The root got brown black and there were no callus appeared.
The most appropriate medium for callus proliferation of leaves were MS+NAA0.2 mg/L +6-BA2.0 mg/L+KT2.0 mg/L.30.0 g/L of sucrose as a suitable carbon source for callus proliferation. CH with 0.4 g/L was good for callus proliferation. The results showed that the growth curve of leaf callus was the S-curve, The grow process can be divided into slow growth phase, exponential phase and stationary phase, The growth period of leaf callus was about 30d.The callus proliferation medium of Stems and petals were same, They were MS+ NAA0.2 mg/L+6-BA3.0 mg/L+sucrose 30.0 g/L+CH0.4 g/L+Agar 7.0 g/L. The medium of anther proliferation were MS+NAA0.2 mg/L+6-BA2.0 mg/L+sucrose 30.0 g/L+CH0.4 g/L+Agar 7.0 g/L.12h light was good for stem proliferation.
The effects of light intensity, anti-oxidants and adsorbents to callus browning were studied in this experiment. The results showed that after dark conditions for 1 week and then cultured in light intensity 19umol/m2/s may be good choice, In this condition,not only ensure the callus proliferation but also greatly reduce the callus browning. Several antioxidants had good effect to callus proliferation and reduced the rate of callus browning. AgNO3 was good for inhibiting the browning. 1.0g/LPVP and 2.0g/L activated carbon were good for inhibiting browning.
Callus structure observation of peony. The result showed that bud primordia are not from the surface of callus but occurred near the surface, and then began to break through the surface cell to grow. Different developmental stages were found in different shapes.
Using scanning electron microscopy, different callus culture occurs at different times different changes can be found, some callus can develop into embryoids, while others are unable to complete normal development. embryogenic callus and non-embryogenic callus may exist on one callus.15-20d after subculture ofth was the fastest time, after this period callus began to grow slow,so after the 15-20d of subculture is more good subculture period.
Callus differentiation of peony. The most suitable medium for callus differentiation of leaves were MS+6-BA2.0 mg/L+NAA0.2 mg/L+TDZ0.3 mg/L+sucrose 30.0 g/L+Agar 7.0 g/L. CH had little effect on the callus differentiation., The highest callus differentiation rate achieved when the sucrose concentration of 30g/L. Activated carbon were negative to callus differentiation, but good for the growth of adventitious buds.
The most appropriate callus differentiation medium of stem were MS+6-BA3.0 mg/L+ NAA0.2 mg/L. That of anther were MS+6-BA2.0 mg/L+NAA0.2 mg/L+KT0.3 mg/L, After dried 3d with filter paper,the anther callus formation quickly. The suitable medium of petal callus differentiation were MS+ZT 0.5 mg/L+6-BA2.0 mg/L.
Root induction and transplanting of peony. Four mediums were used in rooting induction,The results showed that 1/2MSand WPM were appropriate medium.The suitable sucrose concentration was 30.0g/L. The best medium were 1/2MS+NAA0.1mg/L+ IBA0.2mg/L+sucrose 30.0 g/L. Small plants can be transplanted after the domestication of survival, the survival rate was 40.0% or so.
引文
[1]陈有民.园林树木学[M].北京：中国林业出版社,1988
[2]王莲英.中国牡丹品种图志[M].北京：中国林业出版社,1996
[3]陈平平.欧阳修与牡丹.中国园林[J]1998,40(6)：43-45
[4]徐金光,陈相国.荷泽牡丹产业化发展的思考[J]山东林业科技,2001,136(5)：48-49
[5]宋·欧阳修,洛阳牡丹记[M].丛书集成编.1355
[6]宋·陆游,天彭牡丹谱[M]蒋廷锡主编,古今图书集成北京：中华书局
[7]明·薛凤翔,牡丹史[M]李冬生注合肥：安徽人民出版社.1983
[8]清·陈淏子,花镜[M]修订版第2版尹钦恒校注北京：中国农业出版社,1980 94-96
[9]陈俊愉,中国十大名花[M].上海：上海文化出版社.1989
[10]喻衡,菏泽牡丹济南：山东科技出版社.1980
[11]景新民,郑光华.栽培牡丹的种子萌发和储藏特性简报.植物生理学通讯,1995,31(4)：268-270
[12]BartonLV,ChandlerC. Physiological and morphological effects of gibberellic acid on epicotyl dormancy of tree peony. ContribBoyceThompsonInst,1958,19;201-214
[13]郑光华.几种园林植物种子休眠与发芽的研究.园艺学报,1963,2(3)：311-316
[114]郑相穆,周院宝.凤丹种子的休眠和萌发特性.植物生理学通讯,1995,31(4)：260-262
[15]裴利涛,王占营,张俊萍.牡丹繁殖与促成栽培[J]花卉,1999.1：10
[16]曾端香,尹伟伦,赵孝庆,等.牡丹繁殖技术[J]北京林业大学学报,2000,22(3)：90-95
[17].Aoki N,InOue 1.Studies on production of nursery stock in tree peony(J)Effects of bud position of scion.binding material,time.Cultivar and temperature after grafting on graft-take of grafted tree peony Bulleti of the Faculty of Agriculture Shimane University.1992,26:83-89
[18]刘淑敏,王莲英,吴涤新,等.牡丹[M]北京：中国建筑工业出版社.1987
[19]Witkins H F.Halevy A H. Paeonia. CRC Handbook of Flowering IV CRC Press.1985,4:2-4
[20]杨红超,裴冬丽.牡丹种子胚培养研究[J].广西农业科学2006,37(2)：108-110
[21]黄守印.牡丹胚培养与植株再生[J].植物生理学通讯,1987(2)：54-55
[22]周仁超,姚崇怀.紫斑牡丹胚培养与植株再生[J].亚热带植物科学,2001,30(3)：60
[23]曹小勇.濒危植物紫斑牡丹胚离体培养[J].氨基酸和生物资源,2003,25(2)：35-36.
[24]张子学,丁为群,等.凤丹组织培养研究[J].现代中药研究与实践,2004,18(1)：18-2
[25]李志军,刘志国,李红梅.牡丹组培快繁技术研究[J].山东林业科技,2006,3：39-40
[26]李玉龙,吴德玉,潘淑龙,等.牡丹试管苗繁殖技术的研究[J].科学通报,1984(8)：500-50
[27]谢静萱.枯枝牡丹的组织培养[J].植物生理学通讯,1987(2)：53
[28]张龙勃.牡丹组织培养研究初报[J].河南城市园林,1989(10)：21-23
[29]孔祥生,张妙霞.牡丹离体快繁技术研究[J].北方园艺,1998,3(4)：87-89
[30]张桂花,王洪海,王连祥.牡丹组织培养技术研究[J].山东农业科技,2001(5)：16-18.
[31]陈怡平,廉永康,王勋陵.紫斑牡丹休眠地下芽在组织培养条件下的发育研究[J].西北植物学报,2003,23(2)：314-317.
[32]李丽霞,曲复宁,由翠荣,等.应用正交设计方法筛选牡丹愈伤诱导培养基的研究[J].烟台大学学报：自然科学与工程版,2005,18(1)：41-49
[33]何松林,陈笑蕾,陈莉,等.牡丹叶柄离体培养中褐化防止的初步研究[J].河南科学,2005,23(1)：47-50.
[34]王高潮,刘仲健.中国牡丹培育与鉴赏及文化渊源[M].北京：中国林业出版社,2001：119.
[35]时侠清,张子学.凤凰山牡丹药用器官的愈伤组织培养[J].核农学报,2005,19(3)：186-190.
[36]Buchheim,J.A and Meyer,M.M. Micropropagating of Peony(Paeonia spp.)[J].Biotechnology in Agriculture and Forestry, High-tech and Micropropagation Iv,Springer-Verlag, BerlinHeidelberg.1992,20：269-285
[37]贺丹,雷雅凯,李高燕,等.IBA对牡丹太平红,试管苗生根的影响[J].河南科学,2009,27(6)：687-689
[38]高昌勇.不同牡丹外植体诱导愈伤组织的研究[J].安徽农业科学,2007,35(34)：11036；11111
[39]伍成厚,冯毅敏,陈妙贤,等.印度野牡丹茎段的培养与快速繁殖[J].植物生理学通讯,2006,42(6)：1145-1146
[40]贾文庆,王广印,刘宇,等.牡丹种胚离体培养的研究[J].安徽农业科学,2006,34(24)：6441-6444
[41]郎玉涛,罗晓芳.牡丹愈伤组织的诱导及愈伤褐化抑制的研究[J].河南林业科技,2007,27(1)：4-6
[42]Alber,M.R.J.andKunneman,B.P.A.M.Micropropagation of Paeonia[J].Acta Horticulturae1992,314:85-92
[43]Bouza,L.,Jacques,M.,Sotta,B.,etal.The reactivation of tree peony(Paeonia suffruticosa Andr.)vitroplants by chilling is correlated with modifications of abscisic acid, auxin and cytokinin levels[J].Plant Science.1994a,93:153-160
[44]Beruto,M.,Lanteri,L.and Pirtigallo,C. Micropropagation of tree peony [J].Plant Cell Tiss.Org.Cult.2004,79:249-255
[45]Wang,H.F. and Van Standen,J. Establishment of in vitro cultures of tree peonies[J].South African J.Bot.2001.67:358-361
[46]Harris,R.A.,Mantell,S.H. Effect of stage II subculture duration on the multiplication rate and rooting capacity of micropropagated shoots of tree peony[J].J.Hort.Sci.1991,66(l):95-102
[47]Bouza,L. Sotta,B.Bonnet,M.,etal.Hormone content and meristematic activity of Paeonia suffruticosa Andr.cv.'Mme de Vatry'vitroplants during in vitro rooting[J]Acta Horticulturae.1992,302:213-216
[48]Bouza,L.,Jacques,M.,Sotta,B.,etal. The differential effect of N6-benzyl-adenine and N6-(A2-isppentenyl)-adenine on in vitro propagation of Paeonia suffruticosa Andr.is correlated with different hormone contents[J].Plant Cell Report.1993,12:593-596
[49]Bouza,L.,Jacques,M.,Sotta,B.,etal.Relationship between auxin and cytokinin contents and in vitro rooting of tree peony(Paeonia suffruticosa Andr.)[J].Plant Growth Regulation.1994d,15:69-73
[50]Bouza,L.,Jacques,M.and Miginiac,E. Requirements for in vitro rooting of Paeonia suffruticosa Andr.cv. 'Mme de Vatry'[J].Scientia Horticulturae.1994c,58:223-233.
[51]BrukhinVB, BatyginaTB. Embryo culture and somatic embryogenesis in culture of Paeonia anomala[J].Phytomorphology,1994,(34):151-157.
[52]Bouza,L.,Jacques,M.and Miginiac,E. Requirements for in vitro propagation of Paeonia suffruticosa Andr.cv.'Mme de Vatry':developmental effects of exogenous hormones during the multiplication phase[J]. Scientia Horticulturae.1994b,57:241-251
[53]陈笑蕾.牡丹组织培养的初步研究[J].郑州：河南农业大学,2005.
[54]王忠.植物生理学[M].中国农业出版社,1999.
[55]李志军,王国栋,辛淑亮,等.牡丹组织培养快繁新技术[J].莱阳农学院学报(自然科学版)2006,23(2)：122-125,
[56]储成才,李大卫.牡丹组织培养中玻璃化显现的出现及初步观察[J]河南师范大学学报(自然科学版),1992,(1)：70-73.
[57]周俊辉,周家容,曾浩森,等.园艺植物组织培养中的褐化现象及抗褐化研究进展[J].园艺学报,2000,27(增刊)：481-486
[58]曹孜义,刘国民.实用组织培养技术教程[M].兰州：甘肃科学技术出版社,1999
[59]Margherita Beruto, Luca Lanteri, Cristina Portogallo.Micropropagation of tree peony (Paeonia suffruticosa) [J].PlantCell:Tissue and Organ Culture,2004,(2):249-255.
[60]李嘉钰著.中国牡丹与芍药[M].北京：中国林业出版社,1999
[61]高国训.植物组织培养中的褐化问题[J].植物生理学通讯,1999,35(6)：501-506
[62]李艳敏.三个牡丹品种组织培养技术的研究[D].北京林业大学硕士论文,2004
[63]王立浩,张宝玺,郭家珍,等.辣椒花药培养中若干影响因素研究[J]园艺学报,2004,31(2)：199-204
[64]张淑红,王蒂,王清.影响马铃薯叶肉原生质体褐化的因素及硝酸银对其褐化和分裂的作用[J]中国马铃薯,2004,18(2)：77-81.
[65]Goh C.J,Ng S.K, Lakshmanan P.etal. The role of ethylene on direct shoot bud regeneration from mangosteen(Garcinia mangostana L.)leaves cultured in vitro[J].Plant Sci.1997,124:193-202.
[66]Housti F,Coupe M, Auzac J. Effect of ethylene on enzymatic activities involved in the browning of Hevea beasiliensis callus[J].Physiol Plant,1992,86:445-450
[67]Ozden-Tokatli Y, Ozudogru E.A, Akcin A. In vitro response of pistachio nodal explants to silver nitrate.Scientia Horticulturae[J].2005,106:415-426
[68]熊丽,吴丽芳.观赏花卉的组织培养与大规模生产[M].北京：中国化学工业出版社,2003
[69]Debergh P, Harbaoui Y.Mass propagation of globe artichoke:Evaluation of different hypothesis to overcome vitrification with special reference to water potential[J].Physoil Plant.1981,53:181-187
[70]Singha S,Townsend E C,Oberly G H. Relationship between calcium and agar on vitrification and shoot-tip necrosis of quince(Cydonia oblonga Mill.)shoots in vitro.Plant Cell Tissue and Organ Culture.1990,23:135-142
[71]Heloir M C, Kevers C,Hausman J F,etal.Thomas Gaspar Changes in the concentrations of auxin and polyamines during rooting of in vitro propagation walnut shoots[J].Tree Physiology,1996,16:515-519
[72]高彦,白海霞,等.蒸腾抑制剂对组培苗出瓶移栽后成活率的影响[J].植物生理学通讯,1997,33(3)：171-173
[73]丰峰.提高试管苗移栽成活率的技术研究[J].中国南方果树,2001,30(4)：63-64
[74]Yadav JS, Rajam MV. Spatial distribution of free and conjugaged polyamines in leaves of solamum melongena L. associated with differential morphogenetic capacity:efficient somatic embryogenesis with putrescine[J].JExp Bot.1997.48(313):1537-1545.
[75]崔凯荣,刑更生,周功克等.植物植物激素对体细胞胚发生的诱导和调节[J].遗传.2000.22(4)：349-354.
[76]杨玲.花楸种子生物学和体细胞胚发生体系研究.东北林业大学博士学位论文.2007.6.
[77]高莉萍,包满珠.月季的植株再生及遗传转化研究进展[J].植物学通报.2005.22(2)：231-237.
[78]Kintzios S, Manos C, Malcri O.1999.Somatic embryogenesis from mature leaves of rose[J].Plant Cell Rep.18(5):467-472.
[79]郏艳虹,熊庆娥.植物体细胞胚胎发生的研究[J].四川农业大学学报.2003,21(3)：263-266.
[80]齐力旺,韩一凡,李玲,等.2000.华北落叶松体细胞胚发生及遗传转化实验系统的建立[J].实验生物学报.33(4)：357-365.
[81]Stamp J A, Colby S M.1990.Direct shoot organogenesis and plant regeneration from leaves of grape (vitis spp.)[J].Plant Cell, Tissue and Organ Culture.22(2):127-133.
[82]Atree SM, Fowke LC. Embryogeny of gymnosperm, advanced in synthetic seed technology of conifers. Plant Cell Tiss Organ Cult,1993,35:1-35
[83]胡适宜.被子植物胚胎学.北京高等教育出版社.1982
[84]Wetherell D F, Dougall D K.The potassium requirment for growth and embryogenesis in a wild carrot suspension culture. Physiol Plant,1976,37:73-79
[85]王傲雪,李景富.植物体细胞胚状体的诱导研究及应用[J]黑龙江农业科学.1999,2：39-40
[86]Lakshmanan P, Taji A.2000.Somatic embryogenesis in leguminous plants[J]. Plant. Biol.2:136-148.
[87]韩碧文.1989.胚状体发生的细胞学和生理生化研究[J].植物学通报.6(1)：1-4.
[88]Tokuji Y, Kutiyama K. Improvement of gibberellins and cytokinin in the formation of embryogenic cell clumps in carrot (Daucus carota) [J]. Plant Physiol.2003.160:133-141.
[89]Hutchinson M.J., Krishna Raj S. and Saxena P.K. Inhibitory effect of GA3 on the development of Thidiazuron-induced somatic embryogenesis in geranium (Pelargonium x hortorum Bailey)hypocotyls cultures[J]. Plant Cell Rep.1997.16:435-438.
[90]Hita O., Lafarga C. and Guerra H. Somatic embryogenesis from chickpea (Cicer arietinum L.)immature cotyledons:the effect of zeatin, gibberelllic acid and indole-3-butyric acid[J]. Acta Physiol. Plant. 1997.19:333-338.
[91]Rudus L, Kepcrynska E, Kepcrynska J. Regulation of Medicago sativa L. somatic embryogenesis by gibberellins[J]. Plant Growth Regulat.2002.36:91-95.
[92]贺红,潘瑞炽,韩美丽,等.红江橙体胚发生及影响因素的研究[J].广西植物.1998.18(4)：343-346.
[93]Ammirato PV. The regulation of somatic embryos development in plant cell cultures:suspension culture techniques and hormone requirements[J]. Biotechnology.1983.1:68-74.
[94]Chithra M, Martin K, Sunandakumari P, etal. Somatic embryogenesis, encapsulation, and plant regeneration of Rotula aquataca Lour. a rare rhoeophytic woody medicinal plant [J].In Vitro Cellular and Developmental. Biology-plant,2005,41:28-31
[95]Kim T D, Anbazhzgan V R, Park J I. Somatic embryogenesis in Schisandra chinensis (Turcz)Baill. [J]. In vitro cellular and developmental biology-plant,2005,41:253-257
[96]Kaur N, Pati P K, Sharma M, et al. Somatic embryogenesis from inunature zygotic embryos of Rosa bourboniana Desp.[J]. In vitro Cellular and Developmental Biology-plant,2006,42:124-128
[97]Moon H K, Kim Y W, Lee J S, etal. Micropropagation of Kalopanax pious tree via somatic embryogenesis [J]. In vitro cellular and developmental Biology-plant,2005,41:303-306
[98]Sharma P, Koche V, Quraishi A, etal. Somatic embryogenesis in Buchanania lanzan spreng [J]. In vitro cellular and developmental biology-plant,2005,41:645-647
[99]王伟达,李成浩,等.不同植物生长调节物质对杂种落叶松胚性愈伤组织增殖的影响[J].东北林业大学学报,2008,36(9)：5-7
[100]程钰,孟强,等.长俊木瓜体细胞胚胎诱导的初步研究[J].西北林学院学报.2008,23(6)：98-100
[101]田传卫,尚爱芹,张建甫,等.多花蔷薇假珠芽诱导、体细胞胚发生及植株高效再生[J].园艺学报2008,35(3)：403—408
[102]李琰,崔宏安,冯俊涛,等.雷公藤体细胞胚发生及其植株再生[J].林业科学,2008,44(10)：148-152
[103]张存旭,张焕玲,贾小明,等.栓皮栎体胚的增殖、成熟和萌发[J].林业科学.2008,44(6)：39-44
[104]冯新富,谢丽君,陈厚彬,等.香蕉愈伤组织及体细胞胚诱导过程中的组织学观察[J].果树学报2007,24(6)：788-791
[105]兰大伟,刘永立,原田隆.狗枣猕猴桃叶片离体培养的器官、体细胞胚形成与植株再生[J].果树学报2007,24(2)：218-222
[106]何业华,罗吉,吴会桃,等.菠萝叶基愈伤组织诱导体细胞胚[J].果树学报2007,24(1)：59-63
[107]辛伟杰,徐彬,王广东.花烛体细胞胚胎发生及植株再生研究[J].园艺学报2006,33(6)：1281-1286
[108]Yang Y X, Liu G F, Bao M Z. Somatic embryogenesis and plant regeneration from petioles of Parthenocissus tricuspidata Planch. [J].In vitro cellular and developmental biology plant,200742:520-524
[109]Agrawal V,Sardar P R. In vitro regeneration through somatic embryogenesis and organogenesis using cotyledons of Cassia angustifolia vahl. [J]. In Vitro Cellular and Developmental bilology-Plant,2007, 43:585-592
[110]Capuana M, Petrini G Marco A D, etal. Plant regeneration of common ash (Fraxinus excelsior L)by somatic embryogenesis [J]. In vitro cellular and development biology-plant,2007,43:101-110
[111]Kumar V, Ramakrishna A, Ravishankar G A. Influence of different ethyleneinhibitors on somatic embryogenesis and secondary embryogenesis from coffea canephora[J]In Vitro Cellular& Developmental Biology-plant,2007,43:602-607
[112]Steinmacher D A, Cangahuala-Inocente, Clement C R, et al. Somatic embryogenesis from peach palm zygotic embryos [J]. In Vitro Cellular & Developmental Biology plant.,2007,43:124-133
[113]吴雅琴,赵艳华,刘国俭,等.甜樱桃体细胞胚胎发生及植株再生的研究[J].华北农学报.2006.21(增刊)：125-128
[114]李正红,孙振元,刘秀贤,等.地锦体细胞胚胎发生研究[J].林业科学研究2005,18(1)：36-40
[115]Demoise C F, Partanen C R. Effects of subculturing and physical of medium on the nuclear behavior of a plant tissue culture [J]. Am. J. Bot.,1969,56(2):147-152
[116]Sunderland N. Pollen and anther culture. In:Street H E. (ed)Plant Tissue and Cell Culture,1 sted.,1973, 205-239. University of California Press, Berkeley.
[117]Zenkteler M,Misiura E,Ponitka A,Induction of Androgenetic Embryoids in the in vitro Cultured Anthers of Several Species[J].Specialia,1975,15(3):289-291
[118]Roberts M,Sunderland N.Pollen culture of PaeoniaJohn Innes Ann.Rep,1977,68:60-61
[119]何桂梅.牡丹远缘杂交育种及其胚培养与体细胞胚发生的研究[D]北京：北京林业大学,2006
[120]周秀梅.牡丹体细胞胚胎发生研究[D]北京：北京林业大学,2008
[121]李波,董云波,焦德志,等.北海道黄杨叶片愈伤组织形成及细胞学研究[J].种子,2007,26(11)：44-46
[122]叶建伟等.不同生长调节物质对宣木瓜愈伤组织的诱导[J].南京林业大学学报(自然科学 版),2007,31(6)：125-128
[123]杜希华,等.不同外植体和植物激素对银杏愈伤组织诱导和生长的影响[J]山东师范大学学报(自然科学版)2008,23(1)：129-133
[124]师春娟,等.红豆杉愈伤组织诱导试验初探[J]甘肃林业科技,2007,32(4)：19-22
[125]伍成厚,等.红千层愈伤组织诱导及植株再生[J]中南林业科技大学学报,2007,27(6)：104-108
[126]赖明航,等.马尾松愈伤组织诱导条件的研究[J]三峡大学学报(自然科学版),2007,29(5)：457-460
[127]吴延军,张上隆,谢鸣,等.桃幼胚及幼胚子叶再生的研究[J].林业科学,2005,41(9)：45-51
[128]梁巧玲,等.甘蔗愈伤组织诱导与植株再生研究[J]安徽农业科学,2007,35(32)：10246-10247
[129]蒋炜,等.光叶海桐愈伤组织诱导及增殖研究[J]绵阳师范学院学报(自然科学版),2007,26(11)：61-65
[130]龚伟,胡庭兴,宫渊波,等.光叶子花茎段愈伤组织的诱导及其植株再生的研究[J].园艺学报,2005,32(6)：1125-1128
[131]姚丽娟,等.红掌茎段愈伤组织的诱导与植株再生及内生菌的抑制[J]浙江亚热带作物通讯,2006,28(2)：6-14
[132]赵恒田,等.辽东楤木愈伤组织诱导及增殖技术研究[J]农业生物技术科学,2008,24(1)：64-67
[133]张彦波,等.喜树愈伤组织诱导与抑制褐化的研究[J]安徽农业科学,2007,35(36)：11902-11904
[134]樊青峰,等.中国李Nubiana离体叶片愈伤组织诱导的研究[J]华中农业大学学报,2007，10(5)：680-683
[135]姜蕾,兰天维,黎扬辉,等.影响红掌愈伤组织诱导、增殖和芽分化的因素[J].种子,2006,25(11)：26-30
[136]曹新有,李春莲,余玲,等.小麦成熟胚愈伤组织诱导及分化研究[J].西北植物学报,2006：26(11)：2276-2280
[137]贾利敏,傅晓峰,孙国琴,等.燕麦愈伤组织诱导和分化再生影响因素的研究[J].华北农学报,2006,21(4)：31-34
[138]郭英华,张振贤,关秋竹.四川竹根姜胚性愈伤组织诱导和植株再生[J].河北农业大学学报,2005,28(6):36-39
[139]付志惠,李洪林,李琼,等.紫果猕猴桃幼胚愈伤组织诱导及植株再生[J].武汉植物学研究2004,22(5)：459-462
[140]张小红,张红燕,武军,等.TDZ对香椿愈伤组织诱导及芽增殖生长等的影响[J].西北农林科技大学学报(自然科学版),2002,30(5)：35-38
[141]李冬杰,魏景芳,鲁绍伟,等.不同植物植物激素对冬凌草愈伤组织增殖及褐化的影响[J].水土保持研究,2006,13(3)：149-150
[142]郝建平,张江涛.8种甜荞的种子消毒及愈伤组织诱导和增殖条件[J].植物研究,2000,21(1)：65-69
[143]张海兰,林晓佳,赵博光.植物激素对黑松愈伤组织褐变和增殖的作用[J].山东农业大学学报(自然科学版),2002,33(4)：413-417.
[144]周晓鹿,罗晓芳.皂质芦荟诱导愈伤组织影响因素的初步研究[J].辽宁林业科技,2007,2：22-25
[145]黄勇,周吉源.杜仲愈伤组织的诱导及增殖效应[J].华中师范大学学报(自然科学版),2003,31(1)：102-105
[146]刘玉冰,赵听,谭会娟.荒漠植物红砂愈伤组织诱导与增殖研究[J].中国沙漠,2008,28(2)：255-258
[147]Kyeung II Park,Jeong Doo Choi,Sun Jung Eum,Kiu Weon Kim. Effect of Sucrose and Plant Growth Regulator Concentrations on Callus Proliferation and Adventitious Bud Induction from Callus of Lilium Oriental Hybrid Casa Blanca'[J] Journal of the Korean Society for Horticultural Science,2003,44(6):523--528
[148]毛学文,陈荃.不同光质对毛地黄愈伤组织诱导和增殖的效应[J].植物学通报,1997,14(1)：55-56
[149]张真,李胜,李唯,等.不同光质光对葡萄愈伤组织增殖和白藜芦醇含量的影响[J].植物生理学通讯,2008,44(1)：106-108
[150]李玉平,龚宁,王永宏,等.大花金挖耳愈伤组织诱导与增殖[J].西北植物学报,2006,26(6)：1142-1149
[151]何泼,申延,秦俊哲,等杜仲愈伤组织诱导与增殖的研究[J].陕西科技大学学报,2006,21(4)21-25
[152]张继,廖天江,杨宁,等.高乌头愈伤组织诱导及增殖研究[J].西北师范大学学报(自然科学版),2008,44(1)：83-87
[153]王声斌,黄自然,等.影响蕉柑胚性愈伤组织增殖的因素[J].华南农业大学学报,2002,23(1)：23-23
[154]张宝红,刘方,姚长兵,等.糖源对棉花花药培养的影响[J].江西农业大学学报,1998,20(3)：317-321
[155]李亚东,毕光杨.天然汁对绞股蓝愈伤组织增殖的效应[J].湖北大学学报(自然科学版),1999,21(4)：392-394
[156]马海燕,张博,郝兴明,等.新疆苜蓿愈伤组织再生体系建立的研究[J].新疆农业科学 2005,42(1)：19-23
[157]王清连,王敏,师海荣.植物植物激素对棉花体细胞胚胎发生的诱导及调节作用[J].生物技术通讯,2004,15(6)：577-579
[158]Wakita Y,Sasamoto H,Yokota S, Yoshizawa N. Callus proliferation from leaf protoplasts using phenylurea type cytokinin,4-PU in Betula platyphylla var. japonica. In Vitro Cellular & Development Biology. Plant,1995,31(4):252-258
[159]De Silva, A.E.; Kadir, M.A.; Aziz, M.A.; Kadzimin, S. High and Rapid Callus Proliferation of Pineapple (Ananas comosus L.) cv. Moris. The Scientific World Journal,2006,6:169-175
[160]Y. Sahoo, S. K. Pattnaik and P. K. Chand, Plant regeneration from callus cultures of Morus indica L. derived from seedlings and mature plants. Scientia Horticulturae,1997,69(2):85-98
[161]Cai Yunfei, Liu Yanling, Liu Zhenhua,etc.High-frequency embryogenesis and regeneration of plants with high content of gentiopicroside from the Chinese medicinal plant Gentiana straminea Maxim. In Vitro Cellular & Developmental Biology-Plant,2009,45(6):730-739.
[162]郭胜娟,杨春燕,冯玲玲,等.长春花愈伤组织的诱导和增殖[J].华中师范大学学报(自然科学版),2004,38(2)：228-230
[163]陈红,张绿萍.刺梨花药愈伤组织的增殖培养[J].安徽农业科学,2008,36(11)：4413-4414
[164]冯玲玲,范美华,周吉源.丹参愈伤组织的诱导及增殖效应[J].生物学杂志,2004,21(5)：25-27
[165]齐香君,王薇,陈如意.秦艽愈伤组织诱导及增殖研究[J].陕西科技大学学报,2008,26(1)：42-45
[166]杨春燕,刘树楠,周吉源.驱蚊香草愈伤组织的诱导和增殖[J].华中师范大学学报(自然科学版),2006,40(1)：86-89
[167]马俊莲,刘恺,张子德.‘富有’和‘次郎’甜柿叶片再生植株的研究[J].园艺学报,2006,33(5)：1048-1050
[168]陈金慧,施季森,诸葛强,等.杂交鹅掌楸体细胞胚胎发生研究J].林业科学,2003,39(4)：49-53
[169]范小峰,杨颖丽,刘军梅,等.黄花补血草愈伤组织的诱导和植株再生[J].西北植物学报,2007,27(2)：0257-0261
[170]詹亚光,杨传平.白桦愈伤组织的高效诱导和不定芽分化[J].植物生理学通讯,2002,38(2)：111-114
[171]尚爱芹,蔡汉,闫晓洁,等.北海道黄杨下胚轴的离体培养及植株再生[J].中国农业科学2005,38(12)：2502-2507
[172]刘永立,兰大伟,毕静华,等.葛枣猕猴桃组织培养中的器官形成与植株再生[J].果树学报,2005,22(3)：220-223
[173]李毅,何明珠,马海芸.三倍体毛白杨组织脱分化培养与植株体再生[J].植物研究,2002,22(3)：288-291
[174]李登科,杜学海,王永康,等.六月鲜枣愈伤组织诱导及胚状体发生[J].果树学报,2004,21(5)：414-418
[175]何业华,罗吉,吴会桃,等.菠萝叶基愈伤组织诱导体细胞胚[J].果树学报2007,24(1)：59-63
[176]王文星,屈山,曹成有,等.硝酸银对离体培养烟草叶片愈伤组织形成和芽再生及其脯氨酸和丙二醛含量的影响[J].植物生理学通讯,2006,42(4)：668-670
[177]姚丹,郝文媛,沈刚,等.玉米自交系愈伤组织诱导及植株再生研究[J].玉米科学,2007,15(2)：67-72
[178]高莉萍,包满珠.月季‘萨蔓莎’愈伤组织的诱导及植株再生[J].园艺学报,2005,32(3)：534-536
[179]赵成章,吴连斌,杨长登,等.干燥处理对水稻愈伤组织绿苗再生和若干生理生化特性的影响[J].作物学报,997,23(1)：39-43
[180]Ishii Y, Takamura T, Goi M, etal. Callus induction and somatic embryogenesis of Phalaenopsis[J]. Plant Cell Rep,1998,17:446-450.
[181]郝浩永,尉亚辉,王娅宁,等.花生愈伤组织的诱导和再生体系的建立[J].武汉植物学研究2007,25(4)：410-412
[182]马艳红,于卓,李小雷,等.加拿大披碱草与2种国产披碱草杂种F1愈伤组织诱导及植株再生研究[J].西北植物学报,2006,26(9)：1888-1892
[183]邢桂梅,毕晓颖,雷家军.君子兰花器官离体培养[J].园艺学报,2007,34(6)：1563-1568
[184]张俊卫,唐蜻,包满珠.日本结缕草成熟种子愈伤组织诱导及植株再生的影响因子分析[J].中国农业科学,2006,39(2)：368-374
[185]余桂红,马鸿翔,佘建明,等.草坪型高羊茅成熟种子胚性愈伤组织诱导及植株再生[J].江苏农业学报,2004,20(1)：38-43
[186]曾继吾,易干军,张秋明.番木瓜体胚发生及植株再生研究[J].果树学报,2003,20(6)：471-474
[187]吴若菁,阮少宁.百合胚状体形成途径及植株的再生研究[J].福建林学院学报,2003,23(4)：322-325
[188]谢海燕,毛碧增,单兰兰,等.狗牙根颖果胚性愈伤组织的诱导和胚性细胞的超微结构及植株再生[J]. 植物生理与分子生物学学报,2004,30(2)：209-215
[189]胡凤荣,席梦利,刘光欣,等.东方百合的器官发生与体胚发生研究[J].南京林业大学学报,2007,31(2)：5-8
[190]江洪如,余发新,刘腾云.红千层的离体培养及快速繁殖[J].园艺学报,2005,32(3)：541-543
[191]王瑛,周燕,高述民,等.芨芨草愈伤组织器官发生及植株再生[J].植物学通报,2007,24(5)：636-641
[192]曾泉,徐子勤.几种匍匐翦股颖成熟胚愈伤组织诱导及植株再生[J].西北植物学报,2006,26(10)：2033-2038
[193]李瑞芬,张敬原,赵茂林.结缕草愈伤组织诱导及植株再生[J].园艺学报,2003,30(3)：355-357
[194]马骥,乔琦,肖娅萍,等.防风组织培养中畸形胚状体的发生和控制[J].西北植物学报,2005,25(3)552—556
[195]林侃,王晓军,赵民安,等.新疆天山雪莲体胚诱导与分化研究[J].西北植物学报,2006,26(7)：1351-1354
[196]许耀祖,韦彦余,王晓军,等.薰衣草叶片高频再生体系的建立[J].园艺学报,2006,33(1)：182-185
[197]于洋,卫志明.影响小麦成熟胚再生频率因素的研究[J].分子细胞生物学报,2007,40(6)：443-449
[198]迟吉娜,马峙英,韩改英,等.陆地棉组织培养体细胞胚胎发生技术改进[J].棉花学报,2005,17(4)：195-200
[199]高三基,陈如凯,马宏敏.影响籼稻成熟胚愈伤组织植株再生频率的几个因素[J].作物学报,2004,30(12)：1254-1258
[200]田志宏,严寒,李秋杰,等.马蹄金愈伤组织诱导及植株再生研究[J].华中农业大学学报,2003,22(4)：403-407
[201]赵昌杰,马锋旺,徐凌飞.硝酸银对中国李原生质体分离和培养的影响[J].西北农林科技大学学报(自然科学版),2003,31(6)：100-101
[202]肖荷霞,王瑛,高峰,等.外植体及植物激素对SANDITI紫花苜蓿愈伤组织诱导和分化的影响[J].河北农业大学学报,2003,26(4)：47-52
[203]王清连,刘方,周云,等.棉花组织培养直接胚胎发生和植株再生[J].棉花学报,2002,14(6)340-343
[204]邱璐,王波,王志和,等.史密斯桉愈伤组织的诱导及分化[J].东北林业大学学报,2006,34(4)：18-21
[205]王小玲,樊军锋,余发新,等.奥地利黑松组培苗生根技术研究[J].西北林学院学 报,2007,22(1)：63-66
[206]汪灵丹,张日清.大叶榉组培苗生根诱导和移栽试验[J].经济林研究,2008,26(3)：59-63.
[207]金迪,俞红强,义鸣放.二色补血草组培苗生根培养基及移栽基质的筛选[J].河北农业大学学报,2007,30(6)：54-56
[208]王大平,杨玲.猕猴桃组培苗生根培养的研究[J].安徽农业科学,2008,36(21)：8930-8931
[209]Vinod Kumar, H.B. Gururaj, B.C. Narasimha Prasad.etc. Direct shoot organogenesis on shoot apex from seedling explants of Capsicum annuum L[J]. Scientia Horticulturae 2005,106:237-246
[210]Zai Xueming, Pei Qina, Wang Shuwen,etc. The application of beach plum (Prunus maritima) towasteland vegetation recovery in Jiangsu Province, China:Seedling cloning and transplantation[J]. Ecological Engineering,2009,35:591-596.
[211]Yuan Xuejun, Wang Zhiyong, Liu Jianxiu,etc. Development of a plant regeneration system from seed-derived calluses of centipedegrass [Eremochloa ophiuroides (Munro.) Hack][J]. Scientia Horticulturae 2009,120:96-100
[212]Shang Aiqin, Cai han, Yan Xiaojie,etc. Plant Regeneration from In Vitro Cultured Hypocotyl Explants of Euonymus japonicus Cu zhi[J]. Agricultural Sciences in China,2006,5(3):196-201
[213]Lin J J,Thomas J A,Meyer M M. Tissue culture and embryoid formation in Paeonia lactiflora Pall,[J].Am Peony Soc.Bull.1987,263:24-30
[214]Sohn J K, Kim K S, Kim K M.Development of pollen-derived embryos and ploidy level of their regenerated plants in Paeonia Lactiflora Pall[J].Korean J.Plant Tissue Culture,1994,21(4):215-219
[215]Kim Y S,Lee B K.Callus induction and embryogenesis through pollen culture in Paeonia albiflora Pall.[J].Korean J. Plant Tissue Culture,1995,22(1):13-17
[216]Kwon Y S,Shin Y A,Sohn J K.Effect of phenylacetic acid(PAA)on embryo formation in anther and microspore culture of Paeonia lactiflora[J]. Korean J.Plant Biotechnology,2002,29(3):193-198
[217]Kim H M,Shin J H,Sohn J K.Cryopreservation of somatic embryos of the herbaceous peony(Paeonia lactiflora Pall.)by air drying[J].Cryobilogy,2006,53:69-74
[218]Shin J H, Sohn J K, Kim K M, etal. Plant regeneration through somatic embryogenesis from cotyledon of herbaceous peony (Paeonia lactiflora Pall.)[J]. Plant Tissue Culture,1997,24(5):291-294.
[219]Shin J H,Sohn J K,Kim K M,etal.Effect of plant growth regulators on somatic embryogenesis from cotyledon of herbaceous peony(Paeonia lactiflora Pall.)[J]. Korean J. Plant Tissue Culture,1998,25(2):115-118
[220]Partanen C R.Cytological behaviour of plant tissues in vitro as a reflection of potentialities in vivo[J] Plant Tissue Culture,1965,1-9
[221]Gildow F E,Mitchell.J P.Initiation,growth and nuclear characteristics of tissue cultures of Paeonia suffruticosa [J].Physiol Plant,1977,(39):295-298.
[222]陈怡平,丁兰,赵敏桂,等.用紫斑牡丹不同外植体诱导愈伤组织的研究[J].西北师范大学学报(自然科学版),2001,37(3)：66-69.
[223]Biahoua A, Bonneau L. Control of in vitro Somatic embryogenesis of the spindle tree(Euonymus europaeus L) by the sugar type and the osmotic potential of the culture medium[J]Plant Cell Reports, 1999,19:185-190
[224]Zhou S J, Brown D C W. High efficiency plant production of North American ginseng via somatic embryogenesis from cotyledon explants [J]. Plant Cell Rep,2006 (25):166-173
[225]Sujatha M, Prabakaran A J. High frequency embryogenesis in immature zygotic embryos of sunflower[J].Plant Cell, Tiss. Org. Cult,2001,65:23-29.
[226]达克东,张松,李雅志,等.苹果离体叶片培养直接体细胞胚胎发生研究[J]园艺学报,1996,23(3)：241-245
[227]汤浩茹,王水清,任正隆.核桃体细胞胚发生与转基因研究进展[J].林业科学,2000,36(3)：102-110.
[228]Zimmerman J. Lyiv N. Somatic Emhryogenesis:A Model for Early Development in HigherPlants [J]. The Plant Cell,1993,5:1411-1423
[229]Feher A, Pasternak T P, Dudits D. Transition of somatic plant cells to an embryogenicstate [J]. Plant Cell, Tiss. Org. Cult.,2003,74:201-228
[230]Francis D, Sorrell D A. The interface between the cell cycle and plant growth regulators:amini review [J]. Plant Growth Regulation.2001,33:1-12
[231]Gaj M D. Factors influencing somatic embryogenesis induction and plant regeneration with particular reference to Arabidopsis thaliana L Heynh [J]. Plant Growth Regulation.2004,43:27-47
[232]Stasolla C,Yeltng E C. Recent advances in conifer somatic embryogenesis:improving somatic embryo quality [J]. Plant Cell,Tiss. Org. Cult.,2003a,74:15-35
[233]Stasolla C, Vanzyl L, Egertsdotter U.The effects of polyethylene glycol on gene expression of developing White Spruce somatic embryos[J]. Plant Physiology,2003b,131:49-60.
[234]吕守芳,张守攻,齐力旺,等.日本落叶松体细胞胚胎发生的研究[J].林业科学,2005,41(2)：48-52.
[235]齐力旺,韩一凡,韩素英,等.麦芽糖、NAA及ABA对华北落叶松体细胞胚成熟及生根的影响[J].林业科学,2004,40(1)：52-58.
[236]郑相穆,周阮宝,谷丽萍,等.凤丹种子的休眠与萌发特性[J].植物生理学通讯,1995,31(4)：260-262
[237]Beruto,M.,Lanteri,L.and Pirtigallo,C. Micropropagation of tree peony [J].Plant Cell Tiss.Org.Cult.2004,79:249-255.
[238]宋英今,季静,刘海学,等.安祖花愈伤组织诱导及其分化的正交试验设计[J].核农学报2008,22(3)：300-303
[239]施茜,孙振元,卢琦.梭梭(HaloxylOn ammodendron)愈伤组织诱导及植株再生[J].核农学报,2005,19(6)：441-444
[240]杨小玲,侯正仿,季静,等.同组分培养基及培养温度对安祖花叶片愈伤组织诱导率的影响.[J] 沈阳农业大学学报,2008,39(1)：15-18
[241]沈海龙.植物组织培养[M].北京：中国林业出版社,2005
[242]林荣双,王庆华,梁丽琨,等.TDZ诱导花生幼叶的不定芽和体细胞胚胎发生组织细胞学观察[J].植物研究,2002,23：169-172
[243]徐华松,徐九龙,黄学林.TDZ在植物组织培养中的作用[J].广西植物.1996,16(1)：77-80
[244]Ashok KHG,Murthy HN,Pack KY.Embryogenesis and plant. regeneration from anther cultures of Cucumis sativus L[J].Sci Hortic,2003,98 (3):213-222
[245]Chen J T,Chang W C.Efficient plant regeneration through somatic embryogenesis from callus of Oncidium(Orchidaceae)[J].Plant Sci,2000,160:87-93
[246]Murthy BNS,Murch SJ,Saxena PK,Thidiazuron:a potent regulator of in vitro plant morphogenesis[J].In Vitro Cell DevBiol-Plant,1998,34:267-275
[247]赵桂兰,刘艳芝,尹爱平.大豆花药培养中胚状体萌发的研究[J].科学通报.1998,43：1512-1516
[248]陈宗礼,齐向英,张向前,等TDZ和6-BA对枣树继代培养的影响[J]西北农业学报.2006,15(3)：162-165
[249]宋明,汤青林,邹建,等.不同因素对观赏茄花药愈伤组织诱导的影响[J].西南农业大学学报(自然科 学版),2005,27,(4)：534-536.
[250]王延玲,丰震,赵兰勇,等.仙客来花药培养外植体消毒研究[J].山东林业科技,2004,(4)：8-9.
[251]邹金美,张国广,潘一山.羽衣甘蓝花药培养技术体系的研究[J].漳州师范学院学报：自然科学版,2005,18(2)：94-97.
[252]Chen Lin Jiao, Zhu XueYi, Gu Li,et al.Efficient callus induction and plant regeneration from anther of Chinese narcissus(Narcissus tazetta L.var.chinensis Roem)[J].Plant Cell Reports,2005,24(7):401-407.
[253]Han DongSheng; Niimi Y; Nakano M, etal. Regeneration of haploid plants from anther cultures of the Asiatic hybrid lily'Connecticu King'[J]. Plant Cell,Tissue and Organ Culture,1997,47(2)153-158.
[254]段承俐,张智慧,文国松,等.三七花药培养的研究--愈伤组织的诱导[J].云南农业大学学报,2004,19(5)：510-513
[255]王立浩,张宝玺,郭家珍,等.辣椒花药培养中若干影响因素的研究[J].园艺学报,2004,31(2)：199-204.
[256]曹晓燕,王喆之,赵银萍.毛刺槐花药培养及再生植株的获得[J],西北植物学报,2003,23(3)：456-459.
[257]马菊兰,张博.培养基、蔗糖和植物激素对苜蓿花药愈伤组织诱导的影响[J]新疆农业科学2007,44(6)：839-844.
[258]王延玲,丰震,赵兰勇,等.基因型、低温预处理对仙客来花药培养的影响[J]北方园艺2006,(3)：123.
[259]刘国民.花药离体培养中若干问题的研究进展[J].海南大学学报自然科学版,1994,12(3)：253-259.
[260]Xu Wu,Li Ming,Zhang Jing. The variation of endogenous hormones of barley anther in the process of low temperature pretreatment[J]. Actab Genetica Sinica,1997,24(2):165-169.
[261]Huang Tengbo,You Ruilin. A study on the ultra structural changes in tobacco pollens during the course of cold treatment[J].Acta Scientiarura Naturaliura Universitatis Pekinensis,2003,39(6):780-792.
[262]Huang Jianhua,Lu Ruiju, Wang Yifei,Sun Yuefang,Zhou Runmei. Effects of cool pretreatment on level of am ino acid and polyamine content in barley anthers[J]. Scientia Agricultura Sinica,2001,34(6):626-631.
[263]Nedialka Zagorska,Bojan Dimitrov.Induced and drogenesis in alfalfa(Medicago sativa L.)[J].Plant Cell Reports,1995,14 (4):249-252.
[264]司怀军,戴朝曦,于品华,等.菊花幼嫩花瓣愈伤组织的诱导和植株再生[J].甘肃农业大学学报,1998,2：175-177
[265]刘秀莲,吴月燕.金边瑞香花瓣的愈伤组织诱导和植株再生研究[J].浙江万里学院学报.2006,19(2)：87-90
[266]王军娥,巩振辉,李新凤.牡丹愈伤组织诱导与分化技术的优化研究[J].西北农业学报,2008,17(5)：282-286
[267]张宝红,刘方,姚长兵,等.糖源对棉花花药培养的影响[J].江西农业大学学报,1998,20(3)：317-320
[268]李亚东,毕光杨.天然汁对绞股蓝愈伤组织增殖的效应[J].湖北大学学报(自然科学版).1998,21(4)：392-394
[269]郭胜娟,杨春燕,冯玲玲,等.长春花愈伤组织的诱导和增殖[J].华中师范大学学报(自然科学版).2004,38(2)：228-230
[270]陈红,张绿萍.刺梨花药愈伤组织的增殖培养[J]安徽农业科学.2008,36(11)：4413-4414
[271]冯玲玲,范美华,周吉源.丹参愈伤组织的诱导及增殖效应[J].生物学杂志,2004,21(5)：25-27
[272]Tqutorus T E.Fowke L C,Dunstan D I.Somatic embryogenesis in conifers[J]. Canadian Journal of Botany.1991,69:1873-1899.
[273]张真,李胜,李唯,等.不同光质光对葡萄愈伤组织增殖和白藜芦醇含量的影响[J].植物生理学通讯.2008,44(1)：106-108
[274]杨博,韩振海,张永,等.不同光照强度对玫瑰组织培养中初代培养物褐化的影响[J].园艺园林科学,2003,19(6)：194-196.
[275]Wang Q C,etal. Phenol induced browning and establishment of shoot-tip explants of'Fuji'apple and 'Jinhua'pear cultured vitro[J]. J Hort Sci.1994,69:833-839.
[276]蔡金星,等.不同品种梨多酚氧化酶特性及其抑制荆的研究、河北农业技术师范学院学报.1999.13(1)：55-57.
[277]Yang,S.F.and Hoffman,N.E.Ethylene biosynthesis and its regulation in higher plants[J].Annu.Rev.Plant Physiol.1984,35:155-189
[278]Goh C.J,Ng S.K,Lakshmanan P.etal.The role of ethylene on direct shoot bud regeneration from mangosteen(Garcinia mangoseana L.) leaves cultured in vitro[J].Plant Sci.1997,124:193-202
[279]刘用生,李友勇.植物组织培养中活性炭的使用[J].植物生理学通讯,1994,30(4)：214-217
[280]向凤宁,张举仁,陈惠民.1990.玉米愈伤组织的扫描电镜观察.山东大学学报(自然科学版),25(4)：508-514
[281]张宝红,刘方,姚长兵,等.棉花组织培养体细胞胚胎发生的扫描电镜观察.作物学报,2000.26(1)：124-126
[282]滕俊琳,王以秀,薛庆中.水稻花培愈伤组织及其器官发生过程中的扫描电镜观察.华南农业大学学报1992,41-42
[283]R. Li, A. H. Bruneaul and R. Qu. Improved plant regeneration and in vitro somatic embryogenesis of St Augustinegrass[Stenotaphrum secundatum (Walt.) Kuntze] Plant Breeding 2006,125,52-56.
[284]Bronson M R,Dlxon R K. Cultural factors influencing adventitious shoot and plantlet formation from slash pine cotyledons[J]. New Forests,1991,5:277-288
[285]吴月燕,毛军平,周倩倩.路易斯鸢尾组织培养过程中愈伤组织的诱导和芽的分化[J].浙江农业科学.2009,(1)：86-89
[286]王军娥,巩振辉,李新凤.牡丹愈伤组织诱导与分化技术的优化研究[J].西北农业学报,2008,17(5)：282-286
[287]肖兵,梁元冈.几种生长调节物质及有机附加物对菠萝愈伤组织分化影响的细胞学研究[J].华南农业大学学报,1992,13(1)：69-74
[288]兰芹英,李启任,何惠英,等.红掌愈伤组织诱导和芽的分化[J].园艺学报,2003,30(1)：107-109
[2]王莲英.中国牡丹品种图志[M].北京：中国林业出版社,1996
[3]陈平平.欧阳修与牡丹.中国园林[J]1998,40(6)：43-45
[4]徐金光,陈相国.荷泽牡丹产业化发展的思考[J]山东林业科技,2001,136(5)：48-49
[5]宋·欧阳修,洛阳牡丹记[M].丛书集成编.1355
[6]宋·陆游,天彭牡丹谱[M]蒋廷锡主编,古今图书集成北京：中华书局
[7]明·薛凤翔,牡丹史[M]李冬生注合肥：安徽人民出版社.1983
[8]清·陈淏子,花镜[M]修订版第2版尹钦恒校注北京：中国农业出版社,1980 94-96
[9]陈俊愉,中国十大名花[M].上海：上海文化出版社.1989
[10]喻衡,菏泽牡丹济南：山东科技出版社.1980
[11]景新民,郑光华.栽培牡丹的种子萌发和储藏特性简报.植物生理学通讯,1995,31(4)：268-270
[12]BartonLV,ChandlerC. Physiological and morphological effects of gibberellic acid on epicotyl dormancy of tree peony. ContribBoyceThompsonInst,1958,19;201-214
[13]郑光华.几种园林植物种子休眠与发芽的研究.园艺学报,1963,2(3)：311-316
[114]郑相穆,周院宝.凤丹种子的休眠和萌发特性.植物生理学通讯,1995,31(4)：260-262
[15]裴利涛,王占营,张俊萍.牡丹繁殖与促成栽培[J]花卉,1999.1：10
[16]曾端香,尹伟伦,赵孝庆,等.牡丹繁殖技术[J]北京林业大学学报,2000,22(3)：90-95
[17].Aoki N,InOue 1.Studies on production of nursery stock in tree peony(J)Effects of bud position of scion.binding material,time.Cultivar and temperature after grafting on graft-take of grafted tree peony Bulleti of the Faculty of Agriculture Shimane University.1992,26:83-89
[18]刘淑敏,王莲英,吴涤新,等.牡丹[M]北京：中国建筑工业出版社.1987
[19]Witkins H F.Halevy A H. Paeonia. CRC Handbook of Flowering IV CRC Press.1985,4:2-4
[20]杨红超,裴冬丽.牡丹种子胚培养研究[J].广西农业科学2006,37(2)：108-110
[21]黄守印.牡丹胚培养与植株再生[J].植物生理学通讯,1987(2)：54-55
[22]周仁超,姚崇怀.紫斑牡丹胚培养与植株再生[J].亚热带植物科学,2001,30(3)：60
[23]曹小勇.濒危植物紫斑牡丹胚离体培养[J].氨基酸和生物资源,2003,25(2)：35-36.
[24]张子学,丁为群,等.凤丹组织培养研究[J].现代中药研究与实践,2004,18(1)：18-2
[25]李志军,刘志国,李红梅.牡丹组培快繁技术研究[J].山东林业科技,2006,3：39-40
[26]李玉龙,吴德玉,潘淑龙,等.牡丹试管苗繁殖技术的研究[J].科学通报,1984(8)：500-50
[27]谢静萱.枯枝牡丹的组织培养[J].植物生理学通讯,1987(2)：53
[28]张龙勃.牡丹组织培养研究初报[J].河南城市园林,1989(10)：21-23
[29]孔祥生,张妙霞.牡丹离体快繁技术研究[J].北方园艺,1998,3(4)：87-89
[30]张桂花,王洪海,王连祥.牡丹组织培养技术研究[J].山东农业科技,2001(5)：16-18.
[31]陈怡平,廉永康,王勋陵.紫斑牡丹休眠地下芽在组织培养条件下的发育研究[J].西北植物学报,2003,23(2)：314-317.
[32]李丽霞,曲复宁,由翠荣,等.应用正交设计方法筛选牡丹愈伤诱导培养基的研究[J].烟台大学学报：自然科学与工程版,2005,18(1)：41-49
[33]何松林,陈笑蕾,陈莉,等.牡丹叶柄离体培养中褐化防止的初步研究[J].河南科学,2005,23(1)：47-50.
[34]王高潮,刘仲健.中国牡丹培育与鉴赏及文化渊源[M].北京：中国林业出版社,2001：119.
[35]时侠清,张子学.凤凰山牡丹药用器官的愈伤组织培养[J].核农学报,2005,19(3)：186-190.
[36]Buchheim,J.A and Meyer,M.M. Micropropagating of Peony(Paeonia spp.)[J].Biotechnology in Agriculture and Forestry, High-tech and Micropropagation Iv,Springer-Verlag, BerlinHeidelberg.1992,20：269-285
[37]贺丹,雷雅凯,李高燕,等.IBA对牡丹太平红,试管苗生根的影响[J].河南科学,2009,27(6)：687-689
[38]高昌勇.不同牡丹外植体诱导愈伤组织的研究[J].安徽农业科学,2007,35(34)：11036；11111
[39]伍成厚,冯毅敏,陈妙贤,等.印度野牡丹茎段的培养与快速繁殖[J].植物生理学通讯,2006,42(6)：1145-1146
[40]贾文庆,王广印,刘宇,等.牡丹种胚离体培养的研究[J].安徽农业科学,2006,34(24)：6441-6444
[41]郎玉涛,罗晓芳.牡丹愈伤组织的诱导及愈伤褐化抑制的研究[J].河南林业科技,2007,27(1)：4-6
[42]Alber,M.R.J.andKunneman,B.P.A.M.Micropropagation of Paeonia[J].Acta Horticulturae1992,314:85-92
[43]Bouza,L.,Jacques,M.,Sotta,B.,etal.The reactivation of tree peony(Paeonia suffruticosa Andr.)vitroplants by chilling is correlated with modifications of abscisic acid, auxin and cytokinin levels[J].Plant Science.1994a,93:153-160
[44]Beruto,M.,Lanteri,L.and Pirtigallo,C. Micropropagation of tree peony [J].Plant Cell Tiss.Org.Cult.2004,79:249-255
[45]Wang,H.F. and Van Standen,J. Establishment of in vitro cultures of tree peonies[J].South African J.Bot.2001.67:358-361
[46]Harris,R.A.,Mantell,S.H. Effect of stage II subculture duration on the multiplication rate and rooting capacity of micropropagated shoots of tree peony[J].J.Hort.Sci.1991,66(l):95-102
[47]Bouza,L. Sotta,B.Bonnet,M.,etal.Hormone content and meristematic activity of Paeonia suffruticosa Andr.cv.'Mme de Vatry'vitroplants during in vitro rooting[J]Acta Horticulturae.1992,302:213-216
[48]Bouza,L.,Jacques,M.,Sotta,B.,etal. The differential effect of N6-benzyl-adenine and N6-(A2-isppentenyl)-adenine on in vitro propagation of Paeonia suffruticosa Andr.is correlated with different hormone contents[J].Plant Cell Report.1993,12:593-596
[49]Bouza,L.,Jacques,M.,Sotta,B.,etal.Relationship between auxin and cytokinin contents and in vitro rooting of tree peony(Paeonia suffruticosa Andr.)[J].Plant Growth Regulation.1994d,15:69-73
[50]Bouza,L.,Jacques,M.and Miginiac,E. Requirements for in vitro rooting of Paeonia suffruticosa Andr.cv. 'Mme de Vatry'[J].Scientia Horticulturae.1994c,58:223-233.
[51]BrukhinVB, BatyginaTB. Embryo culture and somatic embryogenesis in culture of Paeonia anomala[J].Phytomorphology,1994,(34):151-157.
[52]Bouza,L.,Jacques,M.and Miginiac,E. Requirements for in vitro propagation of Paeonia suffruticosa Andr.cv.'Mme de Vatry':developmental effects of exogenous hormones during the multiplication phase[J]. Scientia Horticulturae.1994b,57:241-251
[53]陈笑蕾.牡丹组织培养的初步研究[J].郑州：河南农业大学,2005.
[54]王忠.植物生理学[M].中国农业出版社,1999.
[55]李志军,王国栋,辛淑亮,等.牡丹组织培养快繁新技术[J].莱阳农学院学报(自然科学版)2006,23(2)：122-125,
[56]储成才,李大卫.牡丹组织培养中玻璃化显现的出现及初步观察[J]河南师范大学学报(自然科学版),1992,(1)：70-73.
[57]周俊辉,周家容,曾浩森,等.园艺植物组织培养中的褐化现象及抗褐化研究进展[J].园艺学报,2000,27(增刊)：481-486
[58]曹孜义,刘国民.实用组织培养技术教程[M].兰州：甘肃科学技术出版社,1999
[59]Margherita Beruto, Luca Lanteri, Cristina Portogallo.Micropropagation of tree peony (Paeonia suffruticosa) [J].PlantCell:Tissue and Organ Culture,2004,(2):249-255.
[60]李嘉钰著.中国牡丹与芍药[M].北京：中国林业出版社,1999
[61]高国训.植物组织培养中的褐化问题[J].植物生理学通讯,1999,35(6)：501-506
[62]李艳敏.三个牡丹品种组织培养技术的研究[D].北京林业大学硕士论文,2004
[63]王立浩,张宝玺,郭家珍,等.辣椒花药培养中若干影响因素研究[J]园艺学报,2004,31(2)：199-204
[64]张淑红,王蒂,王清.影响马铃薯叶肉原生质体褐化的因素及硝酸银对其褐化和分裂的作用[J]中国马铃薯,2004,18(2)：77-81.
[65]Goh C.J,Ng S.K, Lakshmanan P.etal. The role of ethylene on direct shoot bud regeneration from mangosteen(Garcinia mangostana L.)leaves cultured in vitro[J].Plant Sci.1997,124:193-202.
[66]Housti F,Coupe M, Auzac J. Effect of ethylene on enzymatic activities involved in the browning of Hevea beasiliensis callus[J].Physiol Plant,1992,86:445-450
[67]Ozden-Tokatli Y, Ozudogru E.A, Akcin A. In vitro response of pistachio nodal explants to silver nitrate.Scientia Horticulturae[J].2005,106:415-426
[68]熊丽,吴丽芳.观赏花卉的组织培养与大规模生产[M].北京：中国化学工业出版社,2003
[69]Debergh P, Harbaoui Y.Mass propagation of globe artichoke:Evaluation of different hypothesis to overcome vitrification with special reference to water potential[J].Physoil Plant.1981,53:181-187
[70]Singha S,Townsend E C,Oberly G H. Relationship between calcium and agar on vitrification and shoot-tip necrosis of quince(Cydonia oblonga Mill.)shoots in vitro.Plant Cell Tissue and Organ Culture.1990,23:135-142
[71]Heloir M C, Kevers C,Hausman J F,etal.Thomas Gaspar Changes in the concentrations of auxin and polyamines during rooting of in vitro propagation walnut shoots[J].Tree Physiology,1996,16:515-519
[72]高彦,白海霞,等.蒸腾抑制剂对组培苗出瓶移栽后成活率的影响[J].植物生理学通讯,1997,33(3)：171-173
[73]丰峰.提高试管苗移栽成活率的技术研究[J].中国南方果树,2001,30(4)：63-64
[74]Yadav JS, Rajam MV. Spatial distribution of free and conjugaged polyamines in leaves of solamum melongena L. associated with differential morphogenetic capacity:efficient somatic embryogenesis with putrescine[J].JExp Bot.1997.48(313):1537-1545.
[75]崔凯荣,刑更生,周功克等.植物植物激素对体细胞胚发生的诱导和调节[J].遗传.2000.22(4)：349-354.
[76]杨玲.花楸种子生物学和体细胞胚发生体系研究.东北林业大学博士学位论文.2007.6.
[77]高莉萍,包满珠.月季的植株再生及遗传转化研究进展[J].植物学通报.2005.22(2)：231-237.
[78]Kintzios S, Manos C, Malcri O.1999.Somatic embryogenesis from mature leaves of rose[J].Plant Cell Rep.18(5):467-472.
[79]郏艳虹,熊庆娥.植物体细胞胚胎发生的研究[J].四川农业大学学报.2003,21(3)：263-266.
[80]齐力旺,韩一凡,李玲,等.2000.华北落叶松体细胞胚发生及遗传转化实验系统的建立[J].实验生物学报.33(4)：357-365.
[81]Stamp J A, Colby S M.1990.Direct shoot organogenesis and plant regeneration from leaves of grape (vitis spp.)[J].Plant Cell, Tissue and Organ Culture.22(2):127-133.
[82]Atree SM, Fowke LC. Embryogeny of gymnosperm, advanced in synthetic seed technology of conifers. Plant Cell Tiss Organ Cult,1993,35:1-35
[83]胡适宜.被子植物胚胎学.北京高等教育出版社.1982
[84]Wetherell D F, Dougall D K.The potassium requirment for growth and embryogenesis in a wild carrot suspension culture. Physiol Plant,1976,37:73-79
[85]王傲雪,李景富.植物体细胞胚状体的诱导研究及应用[J]黑龙江农业科学.1999,2：39-40
[86]Lakshmanan P, Taji A.2000.Somatic embryogenesis in leguminous plants[J]. Plant. Biol.2:136-148.
[87]韩碧文.1989.胚状体发生的细胞学和生理生化研究[J].植物学通报.6(1)：1-4.
[88]Tokuji Y, Kutiyama K. Improvement of gibberellins and cytokinin in the formation of embryogenic cell clumps in carrot (Daucus carota) [J]. Plant Physiol.2003.160:133-141.
[89]Hutchinson M.J., Krishna Raj S. and Saxena P.K. Inhibitory effect of GA3 on the development of Thidiazuron-induced somatic embryogenesis in geranium (Pelargonium x hortorum Bailey)hypocotyls cultures[J]. Plant Cell Rep.1997.16:435-438.
[90]Hita O., Lafarga C. and Guerra H. Somatic embryogenesis from chickpea (Cicer arietinum L.)immature cotyledons:the effect of zeatin, gibberelllic acid and indole-3-butyric acid[J]. Acta Physiol. Plant. 1997.19:333-338.
[91]Rudus L, Kepcrynska E, Kepcrynska J. Regulation of Medicago sativa L. somatic embryogenesis by gibberellins[J]. Plant Growth Regulat.2002.36:91-95.
[92]贺红,潘瑞炽,韩美丽,等.红江橙体胚发生及影响因素的研究[J].广西植物.1998.18(4)：343-346.
[93]Ammirato PV. The regulation of somatic embryos development in plant cell cultures:suspension culture techniques and hormone requirements[J]. Biotechnology.1983.1:68-74.
[94]Chithra M, Martin K, Sunandakumari P, etal. Somatic embryogenesis, encapsulation, and plant regeneration of Rotula aquataca Lour. a rare rhoeophytic woody medicinal plant [J].In Vitro Cellular and Developmental. Biology-plant,2005,41:28-31
[95]Kim T D, Anbazhzgan V R, Park J I. Somatic embryogenesis in Schisandra chinensis (Turcz)Baill. [J]. In vitro cellular and developmental biology-plant,2005,41:253-257
[96]Kaur N, Pati P K, Sharma M, et al. Somatic embryogenesis from inunature zygotic embryos of Rosa bourboniana Desp.[J]. In vitro Cellular and Developmental Biology-plant,2006,42:124-128
[97]Moon H K, Kim Y W, Lee J S, etal. Micropropagation of Kalopanax pious tree via somatic embryogenesis [J]. In vitro cellular and developmental Biology-plant,2005,41:303-306
[98]Sharma P, Koche V, Quraishi A, etal. Somatic embryogenesis in Buchanania lanzan spreng [J]. In vitro cellular and developmental biology-plant,2005,41:645-647
[99]王伟达,李成浩,等.不同植物生长调节物质对杂种落叶松胚性愈伤组织增殖的影响[J].东北林业大学学报,2008,36(9)：5-7
[100]程钰,孟强,等.长俊木瓜体细胞胚胎诱导的初步研究[J].西北林学院学报.2008,23(6)：98-100
[101]田传卫,尚爱芹,张建甫,等.多花蔷薇假珠芽诱导、体细胞胚发生及植株高效再生[J].园艺学报2008,35(3)：403—408
[102]李琰,崔宏安,冯俊涛,等.雷公藤体细胞胚发生及其植株再生[J].林业科学,2008,44(10)：148-152
[103]张存旭,张焕玲,贾小明,等.栓皮栎体胚的增殖、成熟和萌发[J].林业科学.2008,44(6)：39-44
[104]冯新富,谢丽君,陈厚彬,等.香蕉愈伤组织及体细胞胚诱导过程中的组织学观察[J].果树学报2007,24(6)：788-791
[105]兰大伟,刘永立,原田隆.狗枣猕猴桃叶片离体培养的器官、体细胞胚形成与植株再生[J].果树学报2007,24(2)：218-222
[106]何业华,罗吉,吴会桃,等.菠萝叶基愈伤组织诱导体细胞胚[J].果树学报2007,24(1)：59-63
[107]辛伟杰,徐彬,王广东.花烛体细胞胚胎发生及植株再生研究[J].园艺学报2006,33(6)：1281-1286
[108]Yang Y X, Liu G F, Bao M Z. Somatic embryogenesis and plant regeneration from petioles of Parthenocissus tricuspidata Planch. [J].In vitro cellular and developmental biology plant,200742:520-524
[109]Agrawal V,Sardar P R. In vitro regeneration through somatic embryogenesis and organogenesis using cotyledons of Cassia angustifolia vahl. [J]. In Vitro Cellular and Developmental bilology-Plant,2007, 43:585-592
[110]Capuana M, Petrini G Marco A D, etal. Plant regeneration of common ash (Fraxinus excelsior L)by somatic embryogenesis [J]. In vitro cellular and development biology-plant,2007,43:101-110
[111]Kumar V, Ramakrishna A, Ravishankar G A. Influence of different ethyleneinhibitors on somatic embryogenesis and secondary embryogenesis from coffea canephora[J]In Vitro Cellular& Developmental Biology-plant,2007,43:602-607
[112]Steinmacher D A, Cangahuala-Inocente, Clement C R, et al. Somatic embryogenesis from peach palm zygotic embryos [J]. In Vitro Cellular & Developmental Biology plant.,2007,43:124-133
[113]吴雅琴,赵艳华,刘国俭,等.甜樱桃体细胞胚胎发生及植株再生的研究[J].华北农学报.2006.21(增刊)：125-128
[114]李正红,孙振元,刘秀贤,等.地锦体细胞胚胎发生研究[J].林业科学研究2005,18(1)：36-40
[115]Demoise C F, Partanen C R. Effects of subculturing and physical of medium on the nuclear behavior of a plant tissue culture [J]. Am. J. Bot.,1969,56(2):147-152
[116]Sunderland N. Pollen and anther culture. In:Street H E. (ed)Plant Tissue and Cell Culture,1 sted.,1973, 205-239. University of California Press, Berkeley.
[117]Zenkteler M,Misiura E,Ponitka A,Induction of Androgenetic Embryoids in the in vitro Cultured Anthers of Several Species[J].Specialia,1975,15(3):289-291
[118]Roberts M,Sunderland N.Pollen culture of PaeoniaJohn Innes Ann.Rep,1977,68:60-61
[119]何桂梅.牡丹远缘杂交育种及其胚培养与体细胞胚发生的研究[D]北京：北京林业大学,2006
[120]周秀梅.牡丹体细胞胚胎发生研究[D]北京：北京林业大学,2008
[121]李波,董云波,焦德志,等.北海道黄杨叶片愈伤组织形成及细胞学研究[J].种子,2007,26(11)：44-46
[122]叶建伟等.不同生长调节物质对宣木瓜愈伤组织的诱导[J].南京林业大学学报(自然科学 版),2007,31(6)：125-128
[123]杜希华,等.不同外植体和植物激素对银杏愈伤组织诱导和生长的影响[J]山东师范大学学报(自然科学版)2008,23(1)：129-133
[124]师春娟,等.红豆杉愈伤组织诱导试验初探[J]甘肃林业科技,2007,32(4)：19-22
[125]伍成厚,等.红千层愈伤组织诱导及植株再生[J]中南林业科技大学学报,2007,27(6)：104-108
[126]赖明航,等.马尾松愈伤组织诱导条件的研究[J]三峡大学学报(自然科学版),2007,29(5)：457-460
[127]吴延军,张上隆,谢鸣,等.桃幼胚及幼胚子叶再生的研究[J].林业科学,2005,41(9)：45-51
[128]梁巧玲,等.甘蔗愈伤组织诱导与植株再生研究[J]安徽农业科学,2007,35(32)：10246-10247
[129]蒋炜,等.光叶海桐愈伤组织诱导及增殖研究[J]绵阳师范学院学报(自然科学版),2007,26(11)：61-65
[130]龚伟,胡庭兴,宫渊波,等.光叶子花茎段愈伤组织的诱导及其植株再生的研究[J].园艺学报,2005,32(6)：1125-1128
[131]姚丽娟,等.红掌茎段愈伤组织的诱导与植株再生及内生菌的抑制[J]浙江亚热带作物通讯,2006,28(2)：6-14
[132]赵恒田,等.辽东楤木愈伤组织诱导及增殖技术研究[J]农业生物技术科学,2008,24(1)：64-67
[133]张彦波,等.喜树愈伤组织诱导与抑制褐化的研究[J]安徽农业科学,2007,35(36)：11902-11904
[134]樊青峰,等.中国李Nubiana离体叶片愈伤组织诱导的研究[J]华中农业大学学报,2007，10(5)：680-683
[135]姜蕾,兰天维,黎扬辉,等.影响红掌愈伤组织诱导、增殖和芽分化的因素[J].种子,2006,25(11)：26-30
[136]曹新有,李春莲,余玲,等.小麦成熟胚愈伤组织诱导及分化研究[J].西北植物学报,2006：26(11)：2276-2280
[137]贾利敏,傅晓峰,孙国琴,等.燕麦愈伤组织诱导和分化再生影响因素的研究[J].华北农学报,2006,21(4)：31-34
[138]郭英华,张振贤,关秋竹.四川竹根姜胚性愈伤组织诱导和植株再生[J].河北农业大学学报,2005,28(6):36-39
[139]付志惠,李洪林,李琼,等.紫果猕猴桃幼胚愈伤组织诱导及植株再生[J].武汉植物学研究2004,22(5)：459-462
[140]张小红,张红燕,武军,等.TDZ对香椿愈伤组织诱导及芽增殖生长等的影响[J].西北农林科技大学学报(自然科学版),2002,30(5)：35-38
[141]李冬杰,魏景芳,鲁绍伟,等.不同植物植物激素对冬凌草愈伤组织增殖及褐化的影响[J].水土保持研究,2006,13(3)：149-150
[142]郝建平,张江涛.8种甜荞的种子消毒及愈伤组织诱导和增殖条件[J].植物研究,2000,21(1)：65-69
[143]张海兰,林晓佳,赵博光.植物激素对黑松愈伤组织褐变和增殖的作用[J].山东农业大学学报(自然科学版),2002,33(4)：413-417.
[144]周晓鹿,罗晓芳.皂质芦荟诱导愈伤组织影响因素的初步研究[J].辽宁林业科技,2007,2：22-25
[145]黄勇,周吉源.杜仲愈伤组织的诱导及增殖效应[J].华中师范大学学报(自然科学版),2003,31(1)：102-105
[146]刘玉冰,赵听,谭会娟.荒漠植物红砂愈伤组织诱导与增殖研究[J].中国沙漠,2008,28(2)：255-258
[147]Kyeung II Park,Jeong Doo Choi,Sun Jung Eum,Kiu Weon Kim. Effect of Sucrose and Plant Growth Regulator Concentrations on Callus Proliferation and Adventitious Bud Induction from Callus of Lilium Oriental Hybrid Casa Blanca'[J] Journal of the Korean Society for Horticultural Science,2003,44(6):523--528
[148]毛学文,陈荃.不同光质对毛地黄愈伤组织诱导和增殖的效应[J].植物学通报,1997,14(1)：55-56
[149]张真,李胜,李唯,等.不同光质光对葡萄愈伤组织增殖和白藜芦醇含量的影响[J].植物生理学通讯,2008,44(1)：106-108
[150]李玉平,龚宁,王永宏,等.大花金挖耳愈伤组织诱导与增殖[J].西北植物学报,2006,26(6)：1142-1149
[151]何泼,申延,秦俊哲,等杜仲愈伤组织诱导与增殖的研究[J].陕西科技大学学报,2006,21(4)21-25
[152]张继,廖天江,杨宁,等.高乌头愈伤组织诱导及增殖研究[J].西北师范大学学报(自然科学版),2008,44(1)：83-87
[153]王声斌,黄自然,等.影响蕉柑胚性愈伤组织增殖的因素[J].华南农业大学学报,2002,23(1)：23-23
[154]张宝红,刘方,姚长兵,等.糖源对棉花花药培养的影响[J].江西农业大学学报,1998,20(3)：317-321
[155]李亚东,毕光杨.天然汁对绞股蓝愈伤组织增殖的效应[J].湖北大学学报(自然科学版),1999,21(4)：392-394
[156]马海燕,张博,郝兴明,等.新疆苜蓿愈伤组织再生体系建立的研究[J].新疆农业科学 2005,42(1)：19-23
[157]王清连,王敏,师海荣.植物植物激素对棉花体细胞胚胎发生的诱导及调节作用[J].生物技术通讯,2004,15(6)：577-579
[158]Wakita Y,Sasamoto H,Yokota S, Yoshizawa N. Callus proliferation from leaf protoplasts using phenylurea type cytokinin,4-PU in Betula platyphylla var. japonica. In Vitro Cellular & Development Biology. Plant,1995,31(4):252-258
[159]De Silva, A.E.; Kadir, M.A.; Aziz, M.A.; Kadzimin, S. High and Rapid Callus Proliferation of Pineapple (Ananas comosus L.) cv. Moris. The Scientific World Journal,2006,6:169-175
[160]Y. Sahoo, S. K. Pattnaik and P. K. Chand, Plant regeneration from callus cultures of Morus indica L. derived from seedlings and mature plants. Scientia Horticulturae,1997,69(2):85-98
[161]Cai Yunfei, Liu Yanling, Liu Zhenhua,etc.High-frequency embryogenesis and regeneration of plants with high content of gentiopicroside from the Chinese medicinal plant Gentiana straminea Maxim. In Vitro Cellular & Developmental Biology-Plant,2009,45(6):730-739.
[162]郭胜娟,杨春燕,冯玲玲,等.长春花愈伤组织的诱导和增殖[J].华中师范大学学报(自然科学版),2004,38(2)：228-230
[163]陈红,张绿萍.刺梨花药愈伤组织的增殖培养[J].安徽农业科学,2008,36(11)：4413-4414
[164]冯玲玲,范美华,周吉源.丹参愈伤组织的诱导及增殖效应[J].生物学杂志,2004,21(5)：25-27
[165]齐香君,王薇,陈如意.秦艽愈伤组织诱导及增殖研究[J].陕西科技大学学报,2008,26(1)：42-45
[166]杨春燕,刘树楠,周吉源.驱蚊香草愈伤组织的诱导和增殖[J].华中师范大学学报(自然科学版),2006,40(1)：86-89
[167]马俊莲,刘恺,张子德.‘富有’和‘次郎’甜柿叶片再生植株的研究[J].园艺学报,2006,33(5)：1048-1050
[168]陈金慧,施季森,诸葛强,等.杂交鹅掌楸体细胞胚胎发生研究J].林业科学,2003,39(4)：49-53
[169]范小峰,杨颖丽,刘军梅,等.黄花补血草愈伤组织的诱导和植株再生[J].西北植物学报,2007,27(2)：0257-0261
[170]詹亚光,杨传平.白桦愈伤组织的高效诱导和不定芽分化[J].植物生理学通讯,2002,38(2)：111-114
[171]尚爱芹,蔡汉,闫晓洁,等.北海道黄杨下胚轴的离体培养及植株再生[J].中国农业科学2005,38(12)：2502-2507
[172]刘永立,兰大伟,毕静华,等.葛枣猕猴桃组织培养中的器官形成与植株再生[J].果树学报,2005,22(3)：220-223
[173]李毅,何明珠,马海芸.三倍体毛白杨组织脱分化培养与植株体再生[J].植物研究,2002,22(3)：288-291
[174]李登科,杜学海,王永康,等.六月鲜枣愈伤组织诱导及胚状体发生[J].果树学报,2004,21(5)：414-418
[175]何业华,罗吉,吴会桃,等.菠萝叶基愈伤组织诱导体细胞胚[J].果树学报2007,24(1)：59-63
[176]王文星,屈山,曹成有,等.硝酸银对离体培养烟草叶片愈伤组织形成和芽再生及其脯氨酸和丙二醛含量的影响[J].植物生理学通讯,2006,42(4)：668-670
[177]姚丹,郝文媛,沈刚,等.玉米自交系愈伤组织诱导及植株再生研究[J].玉米科学,2007,15(2)：67-72
[178]高莉萍,包满珠.月季‘萨蔓莎’愈伤组织的诱导及植株再生[J].园艺学报,2005,32(3)：534-536
[179]赵成章,吴连斌,杨长登,等.干燥处理对水稻愈伤组织绿苗再生和若干生理生化特性的影响[J].作物学报,997,23(1)：39-43
[180]Ishii Y, Takamura T, Goi M, etal. Callus induction and somatic embryogenesis of Phalaenopsis[J]. Plant Cell Rep,1998,17:446-450.
[181]郝浩永,尉亚辉,王娅宁,等.花生愈伤组织的诱导和再生体系的建立[J].武汉植物学研究2007,25(4)：410-412
[182]马艳红,于卓,李小雷,等.加拿大披碱草与2种国产披碱草杂种F1愈伤组织诱导及植株再生研究[J].西北植物学报,2006,26(9)：1888-1892
[183]邢桂梅,毕晓颖,雷家军.君子兰花器官离体培养[J].园艺学报,2007,34(6)：1563-1568
[184]张俊卫,唐蜻,包满珠.日本结缕草成熟种子愈伤组织诱导及植株再生的影响因子分析[J].中国农业科学,2006,39(2)：368-374
[185]余桂红,马鸿翔,佘建明,等.草坪型高羊茅成熟种子胚性愈伤组织诱导及植株再生[J].江苏农业学报,2004,20(1)：38-43
[186]曾继吾,易干军,张秋明.番木瓜体胚发生及植株再生研究[J].果树学报,2003,20(6)：471-474
[187]吴若菁,阮少宁.百合胚状体形成途径及植株的再生研究[J].福建林学院学报,2003,23(4)：322-325
[188]谢海燕,毛碧增,单兰兰,等.狗牙根颖果胚性愈伤组织的诱导和胚性细胞的超微结构及植株再生[J]. 植物生理与分子生物学学报,2004,30(2)：209-215
[189]胡凤荣,席梦利,刘光欣,等.东方百合的器官发生与体胚发生研究[J].南京林业大学学报,2007,31(2)：5-8
[190]江洪如,余发新,刘腾云.红千层的离体培养及快速繁殖[J].园艺学报,2005,32(3)：541-543
[191]王瑛,周燕,高述民,等.芨芨草愈伤组织器官发生及植株再生[J].植物学通报,2007,24(5)：636-641
[192]曾泉,徐子勤.几种匍匐翦股颖成熟胚愈伤组织诱导及植株再生[J].西北植物学报,2006,26(10)：2033-2038
[193]李瑞芬,张敬原,赵茂林.结缕草愈伤组织诱导及植株再生[J].园艺学报,2003,30(3)：355-357
[194]马骥,乔琦,肖娅萍,等.防风组织培养中畸形胚状体的发生和控制[J].西北植物学报,2005,25(3)552—556
[195]林侃,王晓军,赵民安,等.新疆天山雪莲体胚诱导与分化研究[J].西北植物学报,2006,26(7)：1351-1354
[196]许耀祖,韦彦余,王晓军,等.薰衣草叶片高频再生体系的建立[J].园艺学报,2006,33(1)：182-185
[197]于洋,卫志明.影响小麦成熟胚再生频率因素的研究[J].分子细胞生物学报,2007,40(6)：443-449
[198]迟吉娜,马峙英,韩改英,等.陆地棉组织培养体细胞胚胎发生技术改进[J].棉花学报,2005,17(4)：195-200
[199]高三基,陈如凯,马宏敏.影响籼稻成熟胚愈伤组织植株再生频率的几个因素[J].作物学报,2004,30(12)：1254-1258
[200]田志宏,严寒,李秋杰,等.马蹄金愈伤组织诱导及植株再生研究[J].华中农业大学学报,2003,22(4)：403-407
[201]赵昌杰,马锋旺,徐凌飞.硝酸银对中国李原生质体分离和培养的影响[J].西北农林科技大学学报(自然科学版),2003,31(6)：100-101
[202]肖荷霞,王瑛,高峰,等.外植体及植物激素对SANDITI紫花苜蓿愈伤组织诱导和分化的影响[J].河北农业大学学报,2003,26(4)：47-52
[203]王清连,刘方,周云,等.棉花组织培养直接胚胎发生和植株再生[J].棉花学报,2002,14(6)340-343
[204]邱璐,王波,王志和,等.史密斯桉愈伤组织的诱导及分化[J].东北林业大学学报,2006,34(4)：18-21
[205]王小玲,樊军锋,余发新,等.奥地利黑松组培苗生根技术研究[J].西北林学院学 报,2007,22(1)：63-66
[206]汪灵丹,张日清.大叶榉组培苗生根诱导和移栽试验[J].经济林研究,2008,26(3)：59-63.
[207]金迪,俞红强,义鸣放.二色补血草组培苗生根培养基及移栽基质的筛选[J].河北农业大学学报,2007,30(6)：54-56
[208]王大平,杨玲.猕猴桃组培苗生根培养的研究[J].安徽农业科学,2008,36(21)：8930-8931
[209]Vinod Kumar, H.B. Gururaj, B.C. Narasimha Prasad.etc. Direct shoot organogenesis on shoot apex from seedling explants of Capsicum annuum L[J]. Scientia Horticulturae 2005,106:237-246
[210]Zai Xueming, Pei Qina, Wang Shuwen,etc. The application of beach plum (Prunus maritima) towasteland vegetation recovery in Jiangsu Province, China:Seedling cloning and transplantation[J]. Ecological Engineering,2009,35:591-596.
[211]Yuan Xuejun, Wang Zhiyong, Liu Jianxiu,etc. Development of a plant regeneration system from seed-derived calluses of centipedegrass [Eremochloa ophiuroides (Munro.) Hack][J]. Scientia Horticulturae 2009,120:96-100
[212]Shang Aiqin, Cai han, Yan Xiaojie,etc. Plant Regeneration from In Vitro Cultured Hypocotyl Explants of Euonymus japonicus Cu zhi[J]. Agricultural Sciences in China,2006,5(3):196-201
[213]Lin J J,Thomas J A,Meyer M M. Tissue culture and embryoid formation in Paeonia lactiflora Pall,[J].Am Peony Soc.Bull.1987,263:24-30
[214]Sohn J K, Kim K S, Kim K M.Development of pollen-derived embryos and ploidy level of their regenerated plants in Paeonia Lactiflora Pall[J].Korean J.Plant Tissue Culture,1994,21(4):215-219
[215]Kim Y S,Lee B K.Callus induction and embryogenesis through pollen culture in Paeonia albiflora Pall.[J].Korean J. Plant Tissue Culture,1995,22(1):13-17
[216]Kwon Y S,Shin Y A,Sohn J K.Effect of phenylacetic acid(PAA)on embryo formation in anther and microspore culture of Paeonia lactiflora[J]. Korean J.Plant Biotechnology,2002,29(3):193-198
[217]Kim H M,Shin J H,Sohn J K.Cryopreservation of somatic embryos of the herbaceous peony(Paeonia lactiflora Pall.)by air drying[J].Cryobilogy,2006,53:69-74
[218]Shin J H, Sohn J K, Kim K M, etal. Plant regeneration through somatic embryogenesis from cotyledon of herbaceous peony (Paeonia lactiflora Pall.)[J]. Plant Tissue Culture,1997,24(5):291-294.
[219]Shin J H,Sohn J K,Kim K M,etal.Effect of plant growth regulators on somatic embryogenesis from cotyledon of herbaceous peony(Paeonia lactiflora Pall.)[J]. Korean J. Plant Tissue Culture,1998,25(2):115-118
[220]Partanen C R.Cytological behaviour of plant tissues in vitro as a reflection of potentialities in vivo[J] Plant Tissue Culture,1965,1-9
[221]Gildow F E,Mitchell.J P.Initiation,growth and nuclear characteristics of tissue cultures of Paeonia suffruticosa [J].Physiol Plant,1977,(39):295-298.
[222]陈怡平,丁兰,赵敏桂,等.用紫斑牡丹不同外植体诱导愈伤组织的研究[J].西北师范大学学报(自然科学版),2001,37(3)：66-69.
[223]Biahoua A, Bonneau L. Control of in vitro Somatic embryogenesis of the spindle tree(Euonymus europaeus L) by the sugar type and the osmotic potential of the culture medium[J]Plant Cell Reports, 1999,19:185-190
[224]Zhou S J, Brown D C W. High efficiency plant production of North American ginseng via somatic embryogenesis from cotyledon explants [J]. Plant Cell Rep,2006 (25):166-173
[225]Sujatha M, Prabakaran A J. High frequency embryogenesis in immature zygotic embryos of sunflower[J].Plant Cell, Tiss. Org. Cult,2001,65:23-29.
[226]达克东,张松,李雅志,等.苹果离体叶片培养直接体细胞胚胎发生研究[J]园艺学报,1996,23(3)：241-245
[227]汤浩茹,王水清,任正隆.核桃体细胞胚发生与转基因研究进展[J].林业科学,2000,36(3)：102-110.
[228]Zimmerman J. Lyiv N. Somatic Emhryogenesis:A Model for Early Development in HigherPlants [J]. The Plant Cell,1993,5:1411-1423
[229]Feher A, Pasternak T P, Dudits D. Transition of somatic plant cells to an embryogenicstate [J]. Plant Cell, Tiss. Org. Cult.,2003,74:201-228
[230]Francis D, Sorrell D A. The interface between the cell cycle and plant growth regulators:amini review [J]. Plant Growth Regulation.2001,33:1-12
[231]Gaj M D. Factors influencing somatic embryogenesis induction and plant regeneration with particular reference to Arabidopsis thaliana L Heynh [J]. Plant Growth Regulation.2004,43:27-47
[232]Stasolla C,Yeltng E C. Recent advances in conifer somatic embryogenesis:improving somatic embryo quality [J]. Plant Cell,Tiss. Org. Cult.,2003a,74:15-35
[233]Stasolla C, Vanzyl L, Egertsdotter U.The effects of polyethylene glycol on gene expression of developing White Spruce somatic embryos[J]. Plant Physiology,2003b,131:49-60.
[234]吕守芳,张守攻,齐力旺,等.日本落叶松体细胞胚胎发生的研究[J].林业科学,2005,41(2)：48-52.
[235]齐力旺,韩一凡,韩素英,等.麦芽糖、NAA及ABA对华北落叶松体细胞胚成熟及生根的影响[J].林业科学,2004,40(1)：52-58.
[236]郑相穆,周阮宝,谷丽萍,等.凤丹种子的休眠与萌发特性[J].植物生理学通讯,1995,31(4)：260-262
[237]Beruto,M.,Lanteri,L.and Pirtigallo,C. Micropropagation of tree peony [J].Plant Cell Tiss.Org.Cult.2004,79:249-255.
[238]宋英今,季静,刘海学,等.安祖花愈伤组织诱导及其分化的正交试验设计[J].核农学报2008,22(3)：300-303
[239]施茜,孙振元,卢琦.梭梭(HaloxylOn ammodendron)愈伤组织诱导及植株再生[J].核农学报,2005,19(6)：441-444
[240]杨小玲,侯正仿,季静,等.同组分培养基及培养温度对安祖花叶片愈伤组织诱导率的影响.[J] 沈阳农业大学学报,2008,39(1)：15-18
[241]沈海龙.植物组织培养[M].北京：中国林业出版社,2005
[242]林荣双,王庆华,梁丽琨,等.TDZ诱导花生幼叶的不定芽和体细胞胚胎发生组织细胞学观察[J].植物研究,2002,23：169-172
[243]徐华松,徐九龙,黄学林.TDZ在植物组织培养中的作用[J].广西植物.1996,16(1)：77-80
[244]Ashok KHG,Murthy HN,Pack KY.Embryogenesis and plant. regeneration from anther cultures of Cucumis sativus L[J].Sci Hortic,2003,98 (3):213-222
[245]Chen J T,Chang W C.Efficient plant regeneration through somatic embryogenesis from callus of Oncidium(Orchidaceae)[J].Plant Sci,2000,160:87-93
[246]Murthy BNS,Murch SJ,Saxena PK,Thidiazuron:a potent regulator of in vitro plant morphogenesis[J].In Vitro Cell DevBiol-Plant,1998,34:267-275
[247]赵桂兰,刘艳芝,尹爱平.大豆花药培养中胚状体萌发的研究[J].科学通报.1998,43：1512-1516
[248]陈宗礼,齐向英,张向前,等TDZ和6-BA对枣树继代培养的影响[J]西北农业学报.2006,15(3)：162-165
[249]宋明,汤青林,邹建,等.不同因素对观赏茄花药愈伤组织诱导的影响[J].西南农业大学学报(自然科 学版),2005,27,(4)：534-536.
[250]王延玲,丰震,赵兰勇,等.仙客来花药培养外植体消毒研究[J].山东林业科技,2004,(4)：8-9.
[251]邹金美,张国广,潘一山.羽衣甘蓝花药培养技术体系的研究[J].漳州师范学院学报：自然科学版,2005,18(2)：94-97.
[252]Chen Lin Jiao, Zhu XueYi, Gu Li,et al.Efficient callus induction and plant regeneration from anther of Chinese narcissus(Narcissus tazetta L.var.chinensis Roem)[J].Plant Cell Reports,2005,24(7):401-407.
[253]Han DongSheng; Niimi Y; Nakano M, etal. Regeneration of haploid plants from anther cultures of the Asiatic hybrid lily'Connecticu King'[J]. Plant Cell,Tissue and Organ Culture,1997,47(2)153-158.
[254]段承俐,张智慧,文国松,等.三七花药培养的研究--愈伤组织的诱导[J].云南农业大学学报,2004,19(5)：510-513
[255]王立浩,张宝玺,郭家珍,等.辣椒花药培养中若干影响因素的研究[J].园艺学报,2004,31(2)：199-204.
[256]曹晓燕,王喆之,赵银萍.毛刺槐花药培养及再生植株的获得[J],西北植物学报,2003,23(3)：456-459.
[257]马菊兰,张博.培养基、蔗糖和植物激素对苜蓿花药愈伤组织诱导的影响[J]新疆农业科学2007,44(6)：839-844.
[258]王延玲,丰震,赵兰勇,等.基因型、低温预处理对仙客来花药培养的影响[J]北方园艺2006,(3)：123.
[259]刘国民.花药离体培养中若干问题的研究进展[J].海南大学学报自然科学版,1994,12(3)：253-259.
[260]Xu Wu,Li Ming,Zhang Jing. The variation of endogenous hormones of barley anther in the process of low temperature pretreatment[J]. Actab Genetica Sinica,1997,24(2):165-169.
[261]Huang Tengbo,You Ruilin. A study on the ultra structural changes in tobacco pollens during the course of cold treatment[J].Acta Scientiarura Naturaliura Universitatis Pekinensis,2003,39(6):780-792.
[262]Huang Jianhua,Lu Ruiju, Wang Yifei,Sun Yuefang,Zhou Runmei. Effects of cool pretreatment on level of am ino acid and polyamine content in barley anthers[J]. Scientia Agricultura Sinica,2001,34(6):626-631.
[263]Nedialka Zagorska,Bojan Dimitrov.Induced and drogenesis in alfalfa(Medicago sativa L.)[J].Plant Cell Reports,1995,14 (4):249-252.
[264]司怀军,戴朝曦,于品华,等.菊花幼嫩花瓣愈伤组织的诱导和植株再生[J].甘肃农业大学学报,1998,2：175-177
[265]刘秀莲,吴月燕.金边瑞香花瓣的愈伤组织诱导和植株再生研究[J].浙江万里学院学报.2006,19(2)：87-90
[266]王军娥,巩振辉,李新凤.牡丹愈伤组织诱导与分化技术的优化研究[J].西北农业学报,2008,17(5)：282-286
[267]张宝红,刘方,姚长兵,等.糖源对棉花花药培养的影响[J].江西农业大学学报,1998,20(3)：317-320
[268]李亚东,毕光杨.天然汁对绞股蓝愈伤组织增殖的效应[J].湖北大学学报(自然科学版).1998,21(4)：392-394
[269]郭胜娟,杨春燕,冯玲玲,等.长春花愈伤组织的诱导和增殖[J].华中师范大学学报(自然科学版).2004,38(2)：228-230
[270]陈红,张绿萍.刺梨花药愈伤组织的增殖培养[J]安徽农业科学.2008,36(11)：4413-4414
[271]冯玲玲,范美华,周吉源.丹参愈伤组织的诱导及增殖效应[J].生物学杂志,2004,21(5)：25-27
[272]Tqutorus T E.Fowke L C,Dunstan D I.Somatic embryogenesis in conifers[J]. Canadian Journal of Botany.1991,69:1873-1899.
[273]张真,李胜,李唯,等.不同光质光对葡萄愈伤组织增殖和白藜芦醇含量的影响[J].植物生理学通讯.2008,44(1)：106-108
[274]杨博,韩振海,张永,等.不同光照强度对玫瑰组织培养中初代培养物褐化的影响[J].园艺园林科学,2003,19(6)：194-196.
[275]Wang Q C,etal. Phenol induced browning and establishment of shoot-tip explants of'Fuji'apple and 'Jinhua'pear cultured vitro[J]. J Hort Sci.1994,69:833-839.
[276]蔡金星,等.不同品种梨多酚氧化酶特性及其抑制荆的研究、河北农业技术师范学院学报.1999.13(1)：55-57.
[277]Yang,S.F.and Hoffman,N.E.Ethylene biosynthesis and its regulation in higher plants[J].Annu.Rev.Plant Physiol.1984,35:155-189
[278]Goh C.J,Ng S.K,Lakshmanan P.etal.The role of ethylene on direct shoot bud regeneration from mangosteen(Garcinia mangoseana L.) leaves cultured in vitro[J].Plant Sci.1997,124:193-202
[279]刘用生,李友勇.植物组织培养中活性炭的使用[J].植物生理学通讯,1994,30(4)：214-217
[280]向凤宁,张举仁,陈惠民.1990.玉米愈伤组织的扫描电镜观察.山东大学学报(自然科学版),25(4)：508-514
[281]张宝红,刘方,姚长兵,等.棉花组织培养体细胞胚胎发生的扫描电镜观察.作物学报,2000.26(1)：124-126
[282]滕俊琳,王以秀,薛庆中.水稻花培愈伤组织及其器官发生过程中的扫描电镜观察.华南农业大学学报1992,41-42
[283]R. Li, A. H. Bruneaul and R. Qu. Improved plant regeneration and in vitro somatic embryogenesis of St Augustinegrass[Stenotaphrum secundatum (Walt.) Kuntze] Plant Breeding 2006,125,52-56.
[284]Bronson M R,Dlxon R K. Cultural factors influencing adventitious shoot and plantlet formation from slash pine cotyledons[J]. New Forests,1991,5:277-288
[285]吴月燕,毛军平,周倩倩.路易斯鸢尾组织培养过程中愈伤组织的诱导和芽的分化[J].浙江农业科学.2009,(1)：86-89
[286]王军娥,巩振辉,李新凤.牡丹愈伤组织诱导与分化技术的优化研究[J].西北农业学报,2008,17(5)：282-286
[287]肖兵,梁元冈.几种生长调节物质及有机附加物对菠萝愈伤组织分化影响的细胞学研究[J].华南农业大学学报,1992,13(1)：69-74
[288]兰芹英,李启任,何惠英,等.红掌愈伤组织诱导和芽的分化[J].园艺学报,2003,30(1)：107-109