白皮松和美国黄松组织培养及快繁技术研究
|
摘要
白皮松和美国黄松组织培养和微体快繁研究表明，在离体培养条件下，以白皮松成熟胚为材料，诱导出大量不定芽；以美国黄松成熟胚为外植体获得了再生小植株，生根率提高到15％。初步探索出白皮松、美国黄松微体快繁的有效途径。其主要结论如下：
（1）胚是白皮松(Pinus bangeana)和美国黄松（Pinus ponderosa）微体快繁的最佳外植体；在组织培养前，切除胚根和其相连的部分—下胚轴，外植体的生长和不定芽的分化较好。
（2）白皮松种子在55℃水中浸泡30min，然后用自来水浸泡3-5天；美国黄松种子用55℃水浸泡10min后，用自来水浸泡2-3天。选用0.2％的KMnO4溶液对白皮松种皮消毒30min，对美国黄松种皮消毒10min。对种皮消毒后，去掉种皮，经70%乙醇漂洗20s，0.1%HgCl2消毒8-10min，无菌水冲洗3-4次，于无菌条件下用解剖刀和镊子剥去胚乳，取出成熟胚接种到培养基上，污染率低。
（3）白皮松不定芽的诱导以MS为最佳培养基，不定芽的诱导率达70%以上，继代培养基仍以MS为基本培养基，不定芽的增殖和生长状况良好。
（4）美国黄松不定芽诱导以GD培养基诱导效果最佳；不定芽增殖选用1/2GD和1/2SH培养基，不定芽伸长选用GD和SH培养基；生根培养选用1/2GD和1/2SH，以1/2GD培养基为佳。
（5）白皮松不定芽诱导采用6-BA 3.5mg· L-1和NAA0.4 mg· L-1及IBA0.1 mg· L-1配合，不定芽的诱导率达到70%以上。
（6）美国黄松采用6-BA和NAA两种配合并进一步缩小激素浓度梯度，6-BA 0.5mg· L-1-1.0mg· L-1，NAA0mg· L-1-0.5mg· L-1时，不定芽的诱导率达到60%以上，在继代培养过程中添加GA3，不添加其他生长调节剂，促进了白皮松不定芽的伸长；采用NAA和GA3组合，使美国黄松嫩稍生根率从2%提高到15%。
（7）在适当的培养基和激素配比条件下，适当提高蔗糖浓度，可作为促进白皮松子叶上不定芽发生的一个重要因素。
（8）在初代培养基中添加活性炭不利于不定芽的诱导和分化；继代培养基中加入1％的活性炭，促进了不定芽的生长；在生根培养基中加入适量活性炭能促进根的生长。
（9）美国黄松嫩稍在1/2GD和1/2SH培养基上均能生根，美国黄松的嫩稍在1/2GD培养基上，NAA1.0 mg· L-1，GA30.5 mg· L-1时的生根率最高，达到16.7%。在1/2SH培养基上，当NAA0.5 mg· L-1和GA30.5 mg· L-1时的生根率，与NAA1.0 mg· L-1不加GA3的生根率相当，达到8.3%。
The study on the tissue culture and micropropagation of Pinus bangeana and Pinus ponderosa showed that: under vitro condition，the mature embryo of Pinus bangeana was introduced in large quantity of adventitious buds, the reproduced plantlet was regenerated from the mature embryo of Pinus ponderosa with great amount of rooting rate. The effective way for micropropagation of Pinus Bangeana and Pinus ponderosa was tentatively explored. The main results are as the following:
(1) The embryo is an optimum explant for micropropagation of Pinus bangeana and Pinus ponderosa. The explant growing and adventitious buds differentiation were well when the embryo root was cut before inoculation.
(2) Pinus bangeana seeds were soaked in hot water with 55℃ for 30min, and then soaked in normal water for 3 to 5 days; the seeds of Pinus ponderosa were soaked in hot water with 55℃ for 10min, and then soaked in water for 2 to 3 days; The well-marinated seeds were sterilized by using 0.2% KMnO4 to Pinus bangeana for 30min and to Pinus ponderosa for 10min, then seedcoat was removed. The embryo was soaked in 70% alcohol for 20s, then transferred into 0.1% HgCl2 for 8 to 10min, and washed by sterile water for 4 times; under disinfective condition, the endosperm was striped by using little dissection knife to get mature embryo into adventitious buds medium. The embryo is rarely polluted.
(3) MS is the best adventitious buds inducement medium for Pinus bangeana. The adventitious inductive rates reached to 70%, it is also used into adventitious buds propagation and growing.
(4) GD is a best medium for adventitious buds inducement to Pinus ponderosa, 1/2GD and 1/2SH were a better medium for adventitious propagation, GD and SH were better medium for adventitious growing, 1/2 GD was a better medium for roots inducement among four mediums.
(5) The inductive rate of adventitious buds for Pinus bangeana reaches 70% above by using 6-BA 3.5 mg· L-1combing NAA 0.4 mg· L-1 with IBA 0.1 mg· L-1.
(6) The inductive rate of adventitious buds for Pinus ponderosa reaches 60% above by using 6-BA combing NAA with lower hormone level at 6-BA 0.5—1.0mg·L-1 and NAA 0—0.5mg·L-1; Which could accelerate the adventitious buds elongation for Pinus bangeana by adding GA3 in adventitious growing medium only. The rooting rates of Pinus ponderosa shoots increased from 2% to 15% by combing NAA and GA3 in medium.
(7) The higher sucrose concentration could be an important factor for adventitious buds
inducement and rooting of Pinus bangeana under the proper medium and hormone combination.
(8) Activated carbon isn’t good for adventitious buds inducement and differentiation, the adventitious buds growing can be promoted by adding 0.1% activated carbon in the medium, and the proper concentration of activated carbon can promote root growing.
(9) The roots can be induced from Pinus ponderosa shoots at 1/2GD and 1/2SH medium. The roots inductive rate of Pinus ponderosa reaches 16.7% at 1/2GD medium with NAA 1.0mg·L-1 and GA3 0.5mg·L-1. The roots inductive rate reaches 8.3% at 1/2SH medium with NAA 0.5mg·L-1 and GA3 0.5mg·L-1, or with NAA 1.0mg·L-1 without GA3.
（1）胚是白皮松(Pinus bangeana)和美国黄松（Pinus ponderosa）微体快繁的最佳外植体；在组织培养前，切除胚根和其相连的部分—下胚轴，外植体的生长和不定芽的分化较好。
（2）白皮松种子在55℃水中浸泡30min，然后用自来水浸泡3-5天；美国黄松种子用55℃水浸泡10min后，用自来水浸泡2-3天。选用0.2％的KMnO4溶液对白皮松种皮消毒30min，对美国黄松种皮消毒10min。对种皮消毒后，去掉种皮，经70%乙醇漂洗20s，0.1%HgCl2消毒8-10min，无菌水冲洗3-4次，于无菌条件下用解剖刀和镊子剥去胚乳，取出成熟胚接种到培养基上，污染率低。
（3）白皮松不定芽的诱导以MS为最佳培养基，不定芽的诱导率达70%以上，继代培养基仍以MS为基本培养基，不定芽的增殖和生长状况良好。
（4）美国黄松不定芽诱导以GD培养基诱导效果最佳；不定芽增殖选用1/2GD和1/2SH培养基，不定芽伸长选用GD和SH培养基；生根培养选用1/2GD和1/2SH，以1/2GD培养基为佳。
（5）白皮松不定芽诱导采用6-BA 3.5mg· L-1和NAA0.4 mg· L-1及IBA0.1 mg· L-1配合，不定芽的诱导率达到70%以上。
（6）美国黄松采用6-BA和NAA两种配合并进一步缩小激素浓度梯度，6-BA 0.5mg· L-1-1.0mg· L-1，NAA0mg· L-1-0.5mg· L-1时，不定芽的诱导率达到60%以上，在继代培养过程中添加GA3，不添加其他生长调节剂，促进了白皮松不定芽的伸长；采用NAA和GA3组合，使美国黄松嫩稍生根率从2%提高到15%。
（7）在适当的培养基和激素配比条件下，适当提高蔗糖浓度，可作为促进白皮松子叶上不定芽发生的一个重要因素。
（8）在初代培养基中添加活性炭不利于不定芽的诱导和分化；继代培养基中加入1％的活性炭，促进了不定芽的生长；在生根培养基中加入适量活性炭能促进根的生长。
（9）美国黄松嫩稍在1/2GD和1/2SH培养基上均能生根，美国黄松的嫩稍在1/2GD培养基上，NAA1.0 mg· L-1，GA30.5 mg· L-1时的生根率最高，达到16.7%。在1/2SH培养基上，当NAA0.5 mg· L-1和GA30.5 mg· L-1时的生根率，与NAA1.0 mg· L-1不加GA3的生根率相当，达到8.3%。
The study on the tissue culture and micropropagation of Pinus bangeana and Pinus ponderosa showed that: under vitro condition，the mature embryo of Pinus bangeana was introduced in large quantity of adventitious buds, the reproduced plantlet was regenerated from the mature embryo of Pinus ponderosa with great amount of rooting rate. The effective way for micropropagation of Pinus Bangeana and Pinus ponderosa was tentatively explored. The main results are as the following:
(1) The embryo is an optimum explant for micropropagation of Pinus bangeana and Pinus ponderosa. The explant growing and adventitious buds differentiation were well when the embryo root was cut before inoculation.
(2) Pinus bangeana seeds were soaked in hot water with 55℃ for 30min, and then soaked in normal water for 3 to 5 days; the seeds of Pinus ponderosa were soaked in hot water with 55℃ for 10min, and then soaked in water for 2 to 3 days; The well-marinated seeds were sterilized by using 0.2% KMnO4 to Pinus bangeana for 30min and to Pinus ponderosa for 10min, then seedcoat was removed. The embryo was soaked in 70% alcohol for 20s, then transferred into 0.1% HgCl2 for 8 to 10min, and washed by sterile water for 4 times; under disinfective condition, the endosperm was striped by using little dissection knife to get mature embryo into adventitious buds medium. The embryo is rarely polluted.
(3) MS is the best adventitious buds inducement medium for Pinus bangeana. The adventitious inductive rates reached to 70%, it is also used into adventitious buds propagation and growing.
(4) GD is a best medium for adventitious buds inducement to Pinus ponderosa, 1/2GD and 1/2SH were a better medium for adventitious propagation, GD and SH were better medium for adventitious growing, 1/2 GD was a better medium for roots inducement among four mediums.
(5) The inductive rate of adventitious buds for Pinus bangeana reaches 70% above by using 6-BA 3.5 mg· L-1combing NAA 0.4 mg· L-1 with IBA 0.1 mg· L-1.
(6) The inductive rate of adventitious buds for Pinus ponderosa reaches 60% above by using 6-BA combing NAA with lower hormone level at 6-BA 0.5—1.0mg·L-1 and NAA 0—0.5mg·L-1; Which could accelerate the adventitious buds elongation for Pinus bangeana by adding GA3 in adventitious growing medium only. The rooting rates of Pinus ponderosa shoots increased from 2% to 15% by combing NAA and GA3 in medium.
(7) The higher sucrose concentration could be an important factor for adventitious buds
inducement and rooting of Pinus bangeana under the proper medium and hormone combination.
(8) Activated carbon isn’t good for adventitious buds inducement and differentiation, the adventitious buds growing can be promoted by adding 0.1% activated carbon in the medium, and the proper concentration of activated carbon can promote root growing.
(9) The roots can be induced from Pinus ponderosa shoots at 1/2GD and 1/2SH medium. The roots inductive rate of Pinus ponderosa reaches 16.7% at 1/2GD medium with NAA 1.0mg·L-1 and GA3 0.5mg·L-1. The roots inductive rate reaches 8.3% at 1/2SH medium with NAA 0.5mg·L-1 and GA3 0.5mg·L-1, or with NAA 1.0mg·L-1 without GA3.
引文
[1] 曹孜义,刘国民.实用植物组织培养技术教程.兰州:甘肃科学技术出版社,1996:1-8
[2] 潘瑞帜,董愚得.植物生理学.北京:高等教育出版社,1995:249－251
[3] 罗士韦.植物组织培养研究的现状和展望.科学通报，1963.14（1）：21-27
[4] 罗士韦.植物组织培养在花卉上的应用. 植物生理学通讯,1979,3:1-2
[5] 罗士伟，许智宏.经济植物组织培养.科学出版社，1988，9：105-112
[6] 颜昌敬.植物组织培养手册.上海科学技术出版社，1990
[7] 梁一池,杨华.植物组织培养技术的研究进展.福建林学院学报,2002,22(1):93-96
[8] 何叶华,胡芳名,谢碧霞.经济林木离体培养研究进展.中南林学院学报, 2000, 20(1):31-39
[9] 施季森.迎接21世纪现代林木生物技术育种的挑战.南京林业大学学报，2000，24（1）：6-10
[10] 容世清.桉树无性系-湛江6号的工厂化微繁研究初报.桉树科技, 1995(2):58-59
[11] 王丽娟,田京伟,杨建雄,等.植物组织培养的研究进展.运城高等师范专科学校学报, 1999,17(3):8-11
[12] 苏晓华，张冰玉，黄烈健等.转基因林木研究进展.林业科学研究, 2003，16（1）：95-103
[13] 王敏杰，卢孟柱.林木基因工程育种现状与发展趋势.世界林业研究, 2002，15（3）：7-13
[14] 贺晶，谭晓风，杨伟.林木转基因工程进展.湖南林业科技, 2001，28（4）19-25
[15] 李文安.植物细胞分化及试管植物.见：余叔文主编. 植物生理与分子生物学. 北京：科学出版社，1992.37-52
[16] 陈振光编著.果树组织培养.上海：上海科技出版社，1987
[17] 陈振光.枇杷离体培养研究进展.福建农学院学报,1991,20(4):422-426
[18] 杨增海.园艺植物组织培养. 北京：农业出版社，1984
[19] 樊军锋，李玲. 84K杨叶片离体培养再生的研究。西北林学院学报，2003，18（2）：40－42
[20] 郝贵霞,朱之悌.毛白杨叶片组培再生芽的玻璃化问题探讨.北京林业大学学报,1999,21(1):68-71
[21] 庞惠仙.灰桉的组织培养与快速繁殖.林业科技通讯,1999(11):29
[22] 丁爱萍,王洪范.影响苹果离体培养叶片分化不定芽的因素.中国果树,1996,(4):20-21
[23] 孙清荣.影响苹果叶片不定芽再生的几个因素.落叶果树,1995(4):3-4
[24] 李梦玲,李嘉瑞.梨叶片离体培养和植株再生.中国农学通报,2001,17(2):8-10
[25] 徐凌飞,任小林.八月红梨叶片不定芽诱导研究.西北农林科技大学学报, 2002, 30(4):73-75
[26] 李名扬,罗金华.发根农杆菌对柑桔的离体转化和植株再生作用.西南农业学报,1996,9(1):90-95
[27] 黄文江,刘庆忠.马哈利樱桃叶片再生的研究.落叶果树,2002,34(4):1-3
[28] 刘庆忠,李志强.甜樱桃矮化砧木吉塞拉的离体叶片再生植株的研究.果树学报,2001,18(5):255-257
[29] 孙仲序,刘静.樱桃砧木叶片和叶柄再生体系的构建.生物技术,2002,12(5):14-16
[30] 石福臣,孔瑾.以叶盘为外植体的白桦的再生.植物研究,2001,21(4):611-614
[31] 张红晓,经剑颖.木本植物组织培养技术研究进展.河南科技大学学报(农学版)，2003，23（3）：66-69
[32] 黄学林,李筱菊.高等植物组织离体培养的形态建成及调控.北京:科学出版社,1995
[33] 李浚明.植物组织培养教程.北京:中国农业大学出版社,2002
[34] 崔凯荣,戴若兰.植物体细胞胚发生的分子生物学.北京:科学出版社,2000
[35] 邓成木,刘进平.热带亚热带植物微繁殖.长沙:湖南科学技术出版社,2001
[36] 王季轩,随长琴.苹果砧木组织培养繁殖的研究.中国农业科学,1981,6:33-37
[37] 王蕴珠.葡萄体细胞愈伤组织诱导与植株再生.植物学报,1985,27(6):661-664
[38] 张锡铭.“昆玉”葡萄快速繁殖.云南植物研究,1985,7(2):245-247
[39] 王大元,张进仁.柑桔的人工培养.植物学报,1975,17(2):149-153
[40] 王春林.香椿组织培养与快速繁殖的研究.园艺学报,1994,21(4):406-407
[41] 王景雪.油菜下胚轴高频率植株再生因素研究.作物学报,1997(3):336-339
[42] 王文静，袁道强，高松洁. 植物组织培养的应用现状. 河北师范大学学报（自然科学版），2000，28（3）：137-139
[43] 桂耀林,马成.植物组织培养.北京:科学出版社,1988
[44] 桂耀林.白皮松离体胚子叶的组织分化和不定芽的形成.植物学刊集（第一集），1983，12（1）：91-95
[45] 雷泽勇,周凤艳,周莉.关于离体胚组织培养在植物育种中的应用进展.防护林科技,2001(3):57-59
[46] 夏光敏,陈惠民.石防风悬浮培养中体细胞胚发生各阶段的细胞超显微结构.植物学报,1991,33(6):482-485
[47] 杨丽军,许智宏.悬浮培养的谷子细胞的胚胎发生和植株再生.实验生物学报, 1985,18(4):493-498
[48] 赵泊,程井辰.石刁柏体细胞胚发生的组织学研究.植物学报, 1994,34(7):491-495
[49] 周俊彦.植物体细胞在组织培养中产生胚状体.植物生理学报, 1981,7(4):389-397
[50] 夏镇澳.植物原生质体培养理论和技术研究进展.农业科学集刊, 1995,2:1-6
[51] 彭信海.植物组织培养在林木遗传育种中的应用.经济林研究，1998，16（2）：54-55
[52] 陈金慧，王洪云，诸葛强等.林木体胚发生技术进展.林业科技开发，2000，14（3）：9-11
[53] 黄健秋.马尾松成熟合子胚的体细胞胚胎发生和植株再生.科学通报，1995，40（1）：72-75
[54] 黄健秋，卫志明，许智宏. 马尾松成熟合子胚的体细胞胚胎发生和植株再生.植物学报, 1995，37（4）：289-294
[55] 黄健秋，卫志明.松属树种的组织培养和原生质体培养.植物学报, 1994，11（1）：34-420
[56] 黄健秋，卫志明.针叶树体细胞胚胎发生的进展.植物生理学通讯,1995，31（2）：85-90
[57] 周微，黄健秋，卫志明，许智宏.云南松成熟胚的不定芽诱导及植株再生.植物生理学报, 1995，31（5）：351-353
[58] 唐巍.火炬松胚性愈伤组织诱导和植株再生的研究.林业科学，1998，34（3）：115-119
[59] 唐巍，欧阳藩，郭仲琛.火炬松成熟胚直接器官发生和植株再生.云南植物研究,1997，19（3）：285-288
[60] 唐巍，欧阳藩，郭仲琛.火炬松合子胚愈伤组织器官发生和植株再生.实验生物学报,1998，31（1）：87-91
[61] 李修庆.植物人工种子研究.北京:北京大学出版社,1990:1-18
[62] 沈熙环等.林业科技通讯，1979，（2）：1
[63] 沈熙环.由欧洲赤松（Pinus sylvestris L）胚离体培养出完整小植株的初步研究.林业科学，1982，18（4）：405-408
[64] 黑龙江省林科所.红松樟子松胚培养效果初报.林业科技通讯,1978，（10）：2-3
[65] 康明，田建强，郑均宝.樟子松成熟胚的离体培养与不定芽的形成. 河北林学院学报，1993，8 （4）：273-276
[66] 王怀智.林业科学，1983，19（3）：292
[67] 刑世岩.松属树种细胞、组织和器官培养名录.植物生理学通讯,1990,（1）：75-79
[68] 齐力旺，杨云龙.油松封顶芽的组织培养.植物生理学通讯，1995，31（1）：40-41
[69] 齐力旺，杨云龙，韩素英等.油松的无性系繁殖与离体培养的研究. 林业科技通讯，1996，（11）：12-14
[70] 孙昌祖.油松组织培养的研究.油松体细胞愈伤组织的诱导,内蒙古林学院学报，1984，1：10-11
[71] 郑均宝，潘冬梅，陈正华.油松离体胚子芽的组织分化和无根试管苗的形成.河北林学院学报,1994，（2）：97-101
[72] 梁玉堂，刑世岩.几种影响黑松针束离体培养再生植株的因子.植物生理学通讯,1990，26（3）：31-33
[73] 吴若菁.马尾松离体胚组织培养初报.福建林学院学报，1993，13（1）：98-100
[74] 成小飞，花晓梅，李文钿.马尾松离体培养条件下的微繁殖和菌根的形成. 林业科学研究，1995，8（3）：241-246
[75] 林静芳译.国外林业文摘. 1994，（1）：57
[76] 林静芳译.国外林业文摘. 1994，（3）：56
[77] 彭方仁译.国外林业文摘. 1995，（5）：59
[78] 贾慧君译.国外林业文摘. 1999，（1）：6
[79] 王明庥主编.林木遗传育种学.中国林业出版社，北京，2001年，278-286
[80] 陈正华主编.木本植物组织培养及其应用. 北京：高等教育出版社，1986：24-74
[81] 黄敏仁等.植物生理学通讯，1990，2：75-78
[82] 龚峥. 松科树种的组织培养.广东林业科技,1990，6：29-33
[83] 阙国宁，郭达初，李金田译，树木组织培养.北京：中国林业出版社，1988：59-94
[84] 阙国宁，房建军，葛万川等.火炬松、湿地松、晚松组培繁殖研究.林业科学研究，1997，10（3）：227-232
[85] 奚元龄，颜昌敬编译.植物组织培养手册.第一版 北京：农业出版社，1990：355
[86] 初立业，宏波邵.植物激素在松树离体快速繁殖中的应用.生物技术通报，1995，1：6-10
[87] 张培杲等译.林木和果树生物技术北京：中国林业出版社，1991：256
[88] 翟应昌.植物组织培养的意义和工厂化育苗.广东林业科技,1987，4：41
[89] 树木志编委会.中国主要树种造林技术.北京：中国林业出版社，1983
[90] 顾淑荣，陈振华，朱至清.白皮松和油松雌配子体愈伤组织的诱导和分化.植物学报，1995，37（3）：
217-221
[91] 中国树木志编委会.树木志（上）.北京：中国林业出版社，1980
[92] 华北树木志编写组.华北树木志.北京：中国林业出版社，1984
[93] 吴宗伦.中国松树的分类与分布.植物分类学报，1956，5（3）：131-278
[94] 韩素英，齐力旺，杨云龙等.几种针叶树离体培养条件的研究.林业科技通讯，1995，10：20-21
[95] 普晓兰，林萍，李乡旺.五针白皮松离体胚培养初步研究.西南林学院学报，1997，17（1）：17-20
[96] 张景群，王德祥，张海花. 美国黄松与秦岭主要松属树种叶燃烧性比较.陕西林业科技，1999，4：53-55
[97] 李曙轩，曹寿椿.蔬菜组织培养.上海：上海科学技术出版社，1983：42-46
[98] Abdullah, A. A., Yeoman, M. M., and Grace, J. In vitro adventitious shoot form embryonic and cotyledonary tissues of Pinus brutia. Plant Cell Tissue Organ Culture.1985,5:35-44
[99] Aiteken, J., Horgan, K. J., and Thorpe, T. A., Influence of explant selection on the shoot forming capacity of juvenile tissue of Pinus radiata. Can.J .For .Res.1981,11(1):112-117
[100] Arnold, S. V., and Eriksson, T. In vitro studies of adventitious shoot formation in Pinus contorta .National Research Council of Canada.1981,59: 870
[101] Arnold, S. V., and Eriksson, T. In vitro study of adventitious shoot formation in Pinus contorta.Can J Bot.1981,59:870-874
[102] Bergmann, B. A., and Stomp, A. M. Effect of genotype on in vitro adventitious shoot formation in pinus radiata and correlations between pairs of phenotypic traits during in vitro shoot development. Plant-Cell, Tissue and Organ Culture.1994,39:185-194
[103] Bergmann, B. A., and Stomp, A. M. Effect of genotype on rooting of hypocotyls and in vitro-produced shoots of Pinus radiata. Plant-Cell, Tissue and Organ Culture .1994,39:195-202
[104] Biondi,and Thorpe, T. A.Growth regulator effects metabolite changes and respiration during initiation in cultured cotyledon explants of Pinus radiata.Bot Gaz.1982,143:20-25
[105] Bornman, C. H., and Jansson, E. Organogenesis in culture Pinus sylvestris L. Tissue. Z. P. Flanzenphysiol.1980, 96:1-6
[106] Becwar, M. R., Nagmani, R., and Wann, S. R. Initiation of embryogenic cultures and somation embryo development in lobiolly pine(Pinus taeda).Can J For Res.1990,20:810-817
[107] Bronson, M. R., and Dixon, R. K.Cultural factors influencing adventitious shoot and plantlet formation from slash pine cotybedone .New Forests.1991,5(4):277-288
[108] Chevreau, E., Skirvin, R. and Abu-Qaoud. H., et al. Adventitious shoot regeneration from leaf tissue of pear (pyrus spp) cultivars in vitro. Plant cell report, 1989(7): 688-691
[109] Cocking, E. C., Method for the isolation of plant protoplasts and vacuoles. Nature, 1960, 187-929
[110] Cupta, P. K., and Durzan, D. J. Biotechnology of somatic polyembryogenesis and plantlet regeneration in loblolly pine.Bio/Tech,1987,5:147-151
[111] Constantin, M. J., Henke R R and Wansur M A. Effect of activated carbon on callus growth and shoot organogenesis in tobacco. In vitro, 1977(13):293-296
[112] Chesick, E. E., Mohn, C. A., and Hackett, W. P., Plantlet multiplication from white pine(Pinus strobes L.)embryos in vitro: bud induction and rooting. Plant Cell Tissue Organ Culture.1991,26:107-114
[113] Campbell, R. A., and Durzan, D. J. Induction of multiple buds and needles intissue cultures of Picea glauca.Can J Bot.1975,53:1652-1657
[114] David, T., Webb, and Olga Diaz Santiago.Cytokinin induced bud formation on Caribean Pine(Pinus caribaea morlet) embryos in vitro. Plant Sci Lett .1983,32:17
[115] David, A., and David, H. Manifestations de diverses potentialities organogenes d’organes ou de fragments d’organes de pin martitime(Pinus pinaster Sol.)en culture in vitro.C R Acad Sci.Paris.1977,184:727-630
[116] David, A.，David, H., and Mateille, T. Invitro adventitious budding on Pinus pinaster otyldedons. Physiol Plant.1982,56:102-107
[117] Dunstan, D. I., Bekkaoui, F., and Pilon, M. et al. Effects of abscisic acid and analogues on the maturation of white spruce(picea glauca) somatic embryos.Plant Sci.1988,58:77
[118] Dumas, E., and Monteuuis, O. In vitro rooting of micropropagated shoots from juvenile and mature Pinus pinaster explants-influence of activated charcoal[J]. Plant Cell Tissue Org Cult, 1995(40):231-235
[119] Ebert, A., Taylor, F., and Balke, J. Changes of 6-benzylaminopurine and 2,4-dichlorphenoxyacetic acid concentrations in plant tissue culture media in the presence of activated charcoal. Plant Cell Tissue Org Cult, 1993(33):157-162
[120] Ellis, D., and Bilderback, D.E.Temporal competence of embryonic Pinus ponderosa cotyledons to form multiple buds in vitro.Amer J Bot .1989,76:348-355
[121] Ellis, D., and Bilderback, D. E. Multiple bud formation by cultured embryos of Pinus ponderosa. Plant Physiol.1984,115:201-204
[122] Ellis, D. et al. Plant Cell, Tissue and Organ Culture.1987,11(1):57
[123] Emily, E., Chesick, Carl, A., Mohn, Wesley P. Hackett. Plantet multiplication from white pine(Pinus strobus L.)embryos in vitro: bud induction and rooting.Plant Cell TISSUE and Organ Culture,1991, 26:107-114
[124] Finer, J. J., Kriebel, H. B., and Becwar, M. R. Initiation of embryogenic callus and susension cultures of eastern white pine(Pinus strobes L.). Plant Cell Rep .1989,8:203-206
[125] Farnum, P., Timmis, R., and Kulp, J. L. Biotechnology of forest yield. Science. 1983, 219:694-702
[126] Fowke, L. C., and Attree, S. M. Applied and basic studies of somatic embryogenesis in white spruce(Picea glauca) and black spruce(Picea mariana).In Soh W Y, Liu J R, Kamamine A(eds). Advances in Developmental Biology and Biotechnology of Higher Plants. Publishedby the Korean
Soc Plant Tissue Culture. Korea, 1993,5
[127] Fonda, R. W., Belanger, L. A., and Burley, L. L. Burning characteristics of western conifer needles. Northwest Sci, 1998,72: 1-9
[128] Go, N. E., Glorina, D., Perez-Orozco and Halos, S. C. In vitro response of embryos from different provenances of pinuns caribbaea var. hondurensis Morelet. Plant-Cell, Tissue and Organ Culture.1993,32:1-7
[129] Gupta, P. K., Durzan, D. J. Somatic plyembryogenesis from callus of mature sugar pine embryos. Bio/Tech.1986,4:643-645
[130] Gupta, P. K., and Durzan, D. J. Shoot multiplication from mature trees of Douglas-fir (Pseudotsuga menziesii) and sugar pine (Pinus lambertiana).Plant Cell Rep.1985,4:177-179
[131] Gupta, P. K., and Durzan, D. J. Somatic plyembryogenesis from mature trees of Douglasfir and sugar pine. Plant Cell Rep.1986,4:643-645
[132] Gladfelter, H. J., and Phillips, G. C. Denovo shoot organogenesis of Pinus eldarica in vitro. Plant Cell Rep.1987,6:163-166
[133] Gupta, P. K., and Pullman, G. S. Method for reproducing coniferous plants by somatic embryogenesis using abscisic acid and osmotic potential variation. U. S. Patent No.5,036,007, 1991
[134] Gupta, P. K., and Pullman, G. S. High concentration enrichment of conifer embryonal cells. U. S. patent No.5,041,382,1991
[135] Genys, J. B., and Hegestad, H. E. Susceptibility of different species,clones and strains of pines to acute injury caused by ozone and sulfer dioxide. Plant disease reporter,1978,6(2):687-691
[136] Halhouli, K. A,, Darwish, N. A., and Alldoon, N. M. Effects of pH and inorganic salts on the adsorption of phenol from aqueous systems on activated decoloring charcoal. Separ Sci Technol, 1995(30):3313-3324
[137] Haissig, B. E., Nelson, N. D., and Kidd, G. H. Trends in the use of tissue culture in forest improvement. Bio/Technology. 1987,5:5
[138] Haddock, P. G. Sapling sugar pine grown from excised mature embryos. Jour Forestry. 1954 , 52 : 434-437
[139] Hohtola, A. Seasonal changes in explant viability and contamination of tissue cultures from mature Scots pine Plant-Cell, Tissue and Organ Culture .1988,15:211-222
[140] James, D. J., Passey, A. J., and Rugini, E. Factors affecting high frequency plant regeneration from apple leaf tissue cultured in vitro. Journal of plant physiology, 1989(132): 148-154
[141] Jones, O. P., Effect of benzylandenine on isolated apple shoots. Nature, 1967, 215: 1514-1515
[142] Johansson, L., and Eriksson, T. Induced embryo formation in anther culture of sever anemone species. Physiol Plant, 1977(40):172-174
[143] Johansson, L., Calleberg, E., and Gedin, A. Correlation between activated charcoal, Fe-EDTA and other
organic media ingredients in cultures of anothers of Anemone canadenis. Physiol Plant, 1990(80):243-249
[144] Jiaswal, V. S., and Armin, M. N. In vitro propagation of guava from shoot cultures of mature trees. J Plant Physiol, 1987(130):7-12
[145] Jang, J. C., and Tainter, F. H. Micropropagation of shortleaf, Virginia and loblolly × shortleaf pine hybrids via organogenesis. Plant-Cell, Tissue and Organ Culture .1991,25: 61-67
[146] Jain, S. N., Dong, N., and Newton, R. J. Somatic embrygenesis in slash pine(Pinus elliottii)from immature embryo culture In vitro. Plant Sci.1989,65:233-241
[147] Kaul, K. Plant regeneration from cotyledon-hypocotyl explants of Pinus strobes L. Plant Cell Rep.1987,6:5-7
[148] Kolevska, Pletikapic, Jelaska, S., and Berllak, J. Bud and shoot formation in juvenile tissue culture of pinus nigra. Silvae genetica . 1983 , 32 : 115-119
[149] Lee, K. C., and Ahn, K. Y, A. Studying on the diversifying of plant species for the landscape construction in Korea. Seoul national university journal of agricultural sciences, 1991, 16(1): 67-80
[150] Lin, Y. Q., Wager, M. R., and Heidmann, L. J. In vitro formation of axillary buds by immature shoots of Pinus ponderosa. Plant-Cell, Tissue and Organ Culture.1991,26(3):161-166
[151] Main, M. L., Marin, J. A. Excised roots cultured in vitro. Plant Cell Report. 1998,18(3/4):350
[152] Merkle, S. A. Plant Tissue Culture Concept and laboratory exercise.CRC Press,1996
[153] M’Kada, J., Dorin, N., and Bigot, C. In vitro micropropagation of Arctostuphylos uvaursi L. Sperengel: comparison between two methodologies.Plant Cell Tissue Org Cult, 1991(24):217-222
[154] Mehra, Palta, A., Smeltzer, R. H., and Mott, R. L. Hormonal control of induced organogenesis : Experiments with excised plant parts of loblolly pine.Tappi. 1978, 61:37 ~40
[155] Mott, R. L., and Amerson, H. V. Tissue culture plantlets produced from Pinus monticola embryonic materials. Forest Sci . 1981 , 27 : 299~304
[156] Minocha, S. C.Callus and adventitious shoot formation in excised embryos of white pine(Pinus strobes). Can J Bot.1980,58:366-370
[157] Mazarl, A., and Camm, E. L. Effect of cytokinins on development and photosynthetic polypeptides during organogenesis of Pinus ponderosa Dougl.cytolodons cultured in vitro.Tissue and Organ Culture.1993,33:81-89
[158] Nancy, E. G., Glorinm, D., and Perez, Orzco, et al. In vitro response of embryos form different provenances of pinus caribaec var.boudurensis Morelet. Plant Cell ,Tissue and Organ Culture.1993,32(1):47-53
[159] Proskauer, J., and Berman, R. Agar culture medium modified to approximate soil condition. Nature(London), 1970(227):1161
[161] Patel, K. R., Shekhwat, N. S., Berlyn, G. P., and Thorpe, T, A. Isolation and culture of protoplasts from
cotyledons of pinus coulteri D.Don.Plant-Cell, Tissue and Organ Culture.1984,3:85-90
[161] Patel, K. R., Kim, H. R., and Thorpe, T. A. Plantlet formation in pitch pine by tissue culture methods. Forest Ecol Managenent.1986,15:147-160
[162] Patel, K. R., and Thorpe, T. A. In vitro differention of plantlets from embryonic explants of lodgepole pine (pinuns contorta Dougl. Ex Lound. Plant-Cell, Tissue and Organ Culture.1984,3:131-142
[163] Pulido, C. M,, Harry, I. S., and Thorpe, T. A. Optimization of bud induction in cotyledonary explants of pinus canariensis. Plant-Cell, Tissue and Organ Culture.1992,29:247-255
[164] Pulido, C. M., Harry, I. S., and Thorpe, T. A. Effect of various bud induction treatments on elongation and rooting of adventitious shoots of Canary Island pine (Pinus canariensis) Plant-Cell, Tissue and Organ Culture.1994,39:225-230
[165] Perez, Bermuudez, P. and Sommer, H. E. Factor affecting adventitious and induction in Pinus elliottii Engelm embryos cultured invitro. Plant Cell, Tissue and Organ Culture. 1987,(11):25-35
[166] Pullman, G. S., and Gupta, P. K. Method for reproducing coniferous plants by somatic embryogenesis using absorbent materials in the development stage media. U.S. patent No. 5,034,326,1991
[167] Rancillac, M., Faye, M., and David, A. In vitro rooting of cloned shoots in Pinus pinaster. Physiol Plant.1982,56:97-101
[168] Robert, D. R. Abscisic acid and mannitol promote early development,maturation and storage protein accumilation in somatic embryos of interior spruce. Physiol Plant.1991,83:247
[169] Skoog, F. C. Tsui. Chemical control of growth and bud formation in tobacco stem segments and callus cultured in vitrol. Amer. J. Bot.,1948,(35):782-787
[170] Skoog, F. C. Tsui. Growth substances and the foemation of buds in plant tissue. In:Plant growth substance,1951,p265-285
[171] Steward, F.C., Holsten, J. R. D., and Simth, J. Growth and organized development of cultured cellsⅡ: growth and division of freely suspended cells. Amer J Bot. 1958,（45）:693-703
[172] Sommer, H. E., Brown, C. L., and Kormanik, P. P. Differentiation of plantlets in long-leaf pine(Pinus palustris Mill) tissue cultured in vitro. Bot Gaz.1975,136:196~200
[173] Sommer, H. E., and Brown, C. L. Plantlets formation in pine tissue cultures. Amer J Bot. Suppl. 1974,61:11
[174] Schwarz, O. J., Schlarbaum, S. E., and Beatu, R. M. Plantlet regeneration form mature zygotic embryos of eastern white pine(Pinus strobes L.). Plant Cell Rep.1988,7:174-174
[175] Takebe, T. Regulation of whole plants from isolated mesophyll protoplasts of tobacoo. Nature, 1971,58: 318-320
[176] Tranvan, H. In vitro adventitious bud formation on isolated seedlings of Pinus sylvestris L.Biologia Plantarum(Praha). 979,21(3):230-233
[177] Tuskan, G. A., Sargent, W. A., and Rensem, T. et al. Influence of plant growth regulators, basal media
and Carbohydrate levels on the in vitro development of Pinus ponderosa cotyledon explants.Plant Cell,Tissue and organ culture,1990,20(1): 47-52
[178] Teasale, R. D., Dawson, P. A, and Woolhouse, H. W. Mineral nutrient requirements of a loblolly pine(Pinus taeda) cell suspension culture,Plant Physiol.986,82: 942-945
[179] Tautorus, T. E,, Fowke, L. C., and Dunstan, D. I. Somatic embryogenes is in conifers. Can J Bot,1991,69:1873
[180] Ueda, N., Dewa, N., and Tsuneya, T., et al. The leaf oil components of lacebark pine, Pinus bungeana Zuuc.Chemistry Express,1989,4(12):813-816
[181] Von Arnold, S., and Eriksson, T. Invitro studies of adventitious shoot formation in Pinus contorta .Can J Bot.1981,59:870-874
[182] White, P. R.The cultivation of animal and plant cells.1943
[183] Whitehead, HCM, K. L. Giles Rapid propagation of polar by tissue culture method . NZ J For Sci 1977,(7):40-43
[184] Weatherhead, M. A., Burdon, J., and Henshaw, G. G. Some effects of activated charcoal as an additive to plant tissue culture media. Z pflanzen-physiol,1978(89):141-147
[185] Webb, D. T., and Santiago, O. D. Induced bud formation on Caribbean pine(Pinus caribaea Morelet)embryos in vitro. Plant Sci Lett.1983,32:17-21
[186] Yam, T. Y., Erns, R., and Arditi, J., et al. Charcoal in orchid seed germination and tissue culture media: a review. Lindleyana,1990(5):256-165
[2] 潘瑞帜,董愚得.植物生理学.北京:高等教育出版社,1995:249－251
[3] 罗士韦.植物组织培养研究的现状和展望.科学通报，1963.14（1）：21-27
[4] 罗士韦.植物组织培养在花卉上的应用. 植物生理学通讯,1979,3:1-2
[5] 罗士伟，许智宏.经济植物组织培养.科学出版社，1988，9：105-112
[6] 颜昌敬.植物组织培养手册.上海科学技术出版社，1990
[7] 梁一池,杨华.植物组织培养技术的研究进展.福建林学院学报,2002,22(1):93-96
[8] 何叶华,胡芳名,谢碧霞.经济林木离体培养研究进展.中南林学院学报, 2000, 20(1):31-39
[9] 施季森.迎接21世纪现代林木生物技术育种的挑战.南京林业大学学报，2000，24（1）：6-10
[10] 容世清.桉树无性系-湛江6号的工厂化微繁研究初报.桉树科技, 1995(2):58-59
[11] 王丽娟,田京伟,杨建雄,等.植物组织培养的研究进展.运城高等师范专科学校学报, 1999,17(3):8-11
[12] 苏晓华，张冰玉，黄烈健等.转基因林木研究进展.林业科学研究, 2003，16（1）：95-103
[13] 王敏杰，卢孟柱.林木基因工程育种现状与发展趋势.世界林业研究, 2002，15（3）：7-13
[14] 贺晶，谭晓风，杨伟.林木转基因工程进展.湖南林业科技, 2001，28（4）19-25
[15] 李文安.植物细胞分化及试管植物.见：余叔文主编. 植物生理与分子生物学. 北京：科学出版社，1992.37-52
[16] 陈振光编著.果树组织培养.上海：上海科技出版社，1987
[17] 陈振光.枇杷离体培养研究进展.福建农学院学报,1991,20(4):422-426
[18] 杨增海.园艺植物组织培养. 北京：农业出版社，1984
[19] 樊军锋，李玲. 84K杨叶片离体培养再生的研究。西北林学院学报，2003，18（2）：40－42
[20] 郝贵霞,朱之悌.毛白杨叶片组培再生芽的玻璃化问题探讨.北京林业大学学报,1999,21(1):68-71
[21] 庞惠仙.灰桉的组织培养与快速繁殖.林业科技通讯,1999(11):29
[22] 丁爱萍,王洪范.影响苹果离体培养叶片分化不定芽的因素.中国果树,1996,(4):20-21
[23] 孙清荣.影响苹果叶片不定芽再生的几个因素.落叶果树,1995(4):3-4
[24] 李梦玲,李嘉瑞.梨叶片离体培养和植株再生.中国农学通报,2001,17(2):8-10
[25] 徐凌飞,任小林.八月红梨叶片不定芽诱导研究.西北农林科技大学学报, 2002, 30(4):73-75
[26] 李名扬,罗金华.发根农杆菌对柑桔的离体转化和植株再生作用.西南农业学报,1996,9(1):90-95
[27] 黄文江,刘庆忠.马哈利樱桃叶片再生的研究.落叶果树,2002,34(4):1-3
[28] 刘庆忠,李志强.甜樱桃矮化砧木吉塞拉的离体叶片再生植株的研究.果树学报,2001,18(5):255-257
[29] 孙仲序,刘静.樱桃砧木叶片和叶柄再生体系的构建.生物技术,2002,12(5):14-16
[30] 石福臣,孔瑾.以叶盘为外植体的白桦的再生.植物研究,2001,21(4):611-614
[31] 张红晓,经剑颖.木本植物组织培养技术研究进展.河南科技大学学报(农学版)，2003，23（3）：66-69
[32] 黄学林,李筱菊.高等植物组织离体培养的形态建成及调控.北京:科学出版社,1995
[33] 李浚明.植物组织培养教程.北京:中国农业大学出版社,2002
[34] 崔凯荣,戴若兰.植物体细胞胚发生的分子生物学.北京:科学出版社,2000
[35] 邓成木,刘进平.热带亚热带植物微繁殖.长沙:湖南科学技术出版社,2001
[36] 王季轩,随长琴.苹果砧木组织培养繁殖的研究.中国农业科学,1981,6:33-37
[37] 王蕴珠.葡萄体细胞愈伤组织诱导与植株再生.植物学报,1985,27(6):661-664
[38] 张锡铭.“昆玉”葡萄快速繁殖.云南植物研究,1985,7(2):245-247
[39] 王大元,张进仁.柑桔的人工培养.植物学报,1975,17(2):149-153
[40] 王春林.香椿组织培养与快速繁殖的研究.园艺学报,1994,21(4):406-407
[41] 王景雪.油菜下胚轴高频率植株再生因素研究.作物学报,1997(3):336-339
[42] 王文静，袁道强，高松洁. 植物组织培养的应用现状. 河北师范大学学报（自然科学版），2000，28（3）：137-139
[43] 桂耀林,马成.植物组织培养.北京:科学出版社,1988
[44] 桂耀林.白皮松离体胚子叶的组织分化和不定芽的形成.植物学刊集（第一集），1983，12（1）：91-95
[45] 雷泽勇,周凤艳,周莉.关于离体胚组织培养在植物育种中的应用进展.防护林科技,2001(3):57-59
[46] 夏光敏,陈惠民.石防风悬浮培养中体细胞胚发生各阶段的细胞超显微结构.植物学报,1991,33(6):482-485
[47] 杨丽军,许智宏.悬浮培养的谷子细胞的胚胎发生和植株再生.实验生物学报, 1985,18(4):493-498
[48] 赵泊,程井辰.石刁柏体细胞胚发生的组织学研究.植物学报, 1994,34(7):491-495
[49] 周俊彦.植物体细胞在组织培养中产生胚状体.植物生理学报, 1981,7(4):389-397
[50] 夏镇澳.植物原生质体培养理论和技术研究进展.农业科学集刊, 1995,2:1-6
[51] 彭信海.植物组织培养在林木遗传育种中的应用.经济林研究，1998，16（2）：54-55
[52] 陈金慧，王洪云，诸葛强等.林木体胚发生技术进展.林业科技开发，2000，14（3）：9-11
[53] 黄健秋.马尾松成熟合子胚的体细胞胚胎发生和植株再生.科学通报，1995，40（1）：72-75
[54] 黄健秋，卫志明，许智宏. 马尾松成熟合子胚的体细胞胚胎发生和植株再生.植物学报, 1995，37（4）：289-294
[55] 黄健秋，卫志明.松属树种的组织培养和原生质体培养.植物学报, 1994，11（1）：34-420
[56] 黄健秋，卫志明.针叶树体细胞胚胎发生的进展.植物生理学通讯,1995，31（2）：85-90
[57] 周微，黄健秋，卫志明，许智宏.云南松成熟胚的不定芽诱导及植株再生.植物生理学报, 1995，31（5）：351-353
[58] 唐巍.火炬松胚性愈伤组织诱导和植株再生的研究.林业科学，1998，34（3）：115-119
[59] 唐巍，欧阳藩，郭仲琛.火炬松成熟胚直接器官发生和植株再生.云南植物研究,1997，19（3）：285-288
[60] 唐巍，欧阳藩，郭仲琛.火炬松合子胚愈伤组织器官发生和植株再生.实验生物学报,1998，31（1）：87-91
[61] 李修庆.植物人工种子研究.北京:北京大学出版社,1990:1-18
[62] 沈熙环等.林业科技通讯，1979，（2）：1
[63] 沈熙环.由欧洲赤松（Pinus sylvestris L）胚离体培养出完整小植株的初步研究.林业科学，1982，18（4）：405-408
[64] 黑龙江省林科所.红松樟子松胚培养效果初报.林业科技通讯,1978，（10）：2-3
[65] 康明，田建强，郑均宝.樟子松成熟胚的离体培养与不定芽的形成. 河北林学院学报，1993，8 （4）：273-276
[66] 王怀智.林业科学，1983，19（3）：292
[67] 刑世岩.松属树种细胞、组织和器官培养名录.植物生理学通讯,1990,（1）：75-79
[68] 齐力旺，杨云龙.油松封顶芽的组织培养.植物生理学通讯，1995，31（1）：40-41
[69] 齐力旺，杨云龙，韩素英等.油松的无性系繁殖与离体培养的研究. 林业科技通讯，1996，（11）：12-14
[70] 孙昌祖.油松组织培养的研究.油松体细胞愈伤组织的诱导,内蒙古林学院学报，1984，1：10-11
[71] 郑均宝，潘冬梅，陈正华.油松离体胚子芽的组织分化和无根试管苗的形成.河北林学院学报,1994，（2）：97-101
[72] 梁玉堂，刑世岩.几种影响黑松针束离体培养再生植株的因子.植物生理学通讯,1990，26（3）：31-33
[73] 吴若菁.马尾松离体胚组织培养初报.福建林学院学报，1993，13（1）：98-100
[74] 成小飞，花晓梅，李文钿.马尾松离体培养条件下的微繁殖和菌根的形成. 林业科学研究，1995，8（3）：241-246
[75] 林静芳译.国外林业文摘. 1994，（1）：57
[76] 林静芳译.国外林业文摘. 1994，（3）：56
[77] 彭方仁译.国外林业文摘. 1995，（5）：59
[78] 贾慧君译.国外林业文摘. 1999，（1）：6
[79] 王明庥主编.林木遗传育种学.中国林业出版社，北京，2001年，278-286
[80] 陈正华主编.木本植物组织培养及其应用. 北京：高等教育出版社，1986：24-74
[81] 黄敏仁等.植物生理学通讯，1990，2：75-78
[82] 龚峥. 松科树种的组织培养.广东林业科技,1990，6：29-33
[83] 阙国宁，郭达初，李金田译，树木组织培养.北京：中国林业出版社，1988：59-94
[84] 阙国宁，房建军，葛万川等.火炬松、湿地松、晚松组培繁殖研究.林业科学研究，1997，10（3）：227-232
[85] 奚元龄，颜昌敬编译.植物组织培养手册.第一版 北京：农业出版社，1990：355
[86] 初立业，宏波邵.植物激素在松树离体快速繁殖中的应用.生物技术通报，1995，1：6-10
[87] 张培杲等译.林木和果树生物技术北京：中国林业出版社，1991：256
[88] 翟应昌.植物组织培养的意义和工厂化育苗.广东林业科技,1987，4：41
[89] 树木志编委会.中国主要树种造林技术.北京：中国林业出版社，1983
[90] 顾淑荣，陈振华，朱至清.白皮松和油松雌配子体愈伤组织的诱导和分化.植物学报，1995，37（3）：
217-221
[91] 中国树木志编委会.树木志（上）.北京：中国林业出版社，1980
[92] 华北树木志编写组.华北树木志.北京：中国林业出版社，1984
[93] 吴宗伦.中国松树的分类与分布.植物分类学报，1956，5（3）：131-278
[94] 韩素英，齐力旺，杨云龙等.几种针叶树离体培养条件的研究.林业科技通讯，1995，10：20-21
[95] 普晓兰，林萍，李乡旺.五针白皮松离体胚培养初步研究.西南林学院学报，1997，17（1）：17-20
[96] 张景群，王德祥，张海花. 美国黄松与秦岭主要松属树种叶燃烧性比较.陕西林业科技，1999，4：53-55
[97] 李曙轩，曹寿椿.蔬菜组织培养.上海：上海科学技术出版社，1983：42-46
[98] Abdullah, A. A., Yeoman, M. M., and Grace, J. In vitro adventitious shoot form embryonic and cotyledonary tissues of Pinus brutia. Plant Cell Tissue Organ Culture.1985,5:35-44
[99] Aiteken, J., Horgan, K. J., and Thorpe, T. A., Influence of explant selection on the shoot forming capacity of juvenile tissue of Pinus radiata. Can.J .For .Res.1981,11(1):112-117
[100] Arnold, S. V., and Eriksson, T. In vitro studies of adventitious shoot formation in Pinus contorta .National Research Council of Canada.1981,59: 870
[101] Arnold, S. V., and Eriksson, T. In vitro study of adventitious shoot formation in Pinus contorta.Can J Bot.1981,59:870-874
[102] Bergmann, B. A., and Stomp, A. M. Effect of genotype on in vitro adventitious shoot formation in pinus radiata and correlations between pairs of phenotypic traits during in vitro shoot development. Plant-Cell, Tissue and Organ Culture.1994,39:185-194
[103] Bergmann, B. A., and Stomp, A. M. Effect of genotype on rooting of hypocotyls and in vitro-produced shoots of Pinus radiata. Plant-Cell, Tissue and Organ Culture .1994,39:195-202
[104] Biondi,and Thorpe, T. A.Growth regulator effects metabolite changes and respiration during initiation in cultured cotyledon explants of Pinus radiata.Bot Gaz.1982,143:20-25
[105] Bornman, C. H., and Jansson, E. Organogenesis in culture Pinus sylvestris L. Tissue. Z. P. Flanzenphysiol.1980, 96:1-6
[106] Becwar, M. R., Nagmani, R., and Wann, S. R. Initiation of embryogenic cultures and somation embryo development in lobiolly pine(Pinus taeda).Can J For Res.1990,20:810-817
[107] Bronson, M. R., and Dixon, R. K.Cultural factors influencing adventitious shoot and plantlet formation from slash pine cotybedone .New Forests.1991,5(4):277-288
[108] Chevreau, E., Skirvin, R. and Abu-Qaoud. H., et al. Adventitious shoot regeneration from leaf tissue of pear (pyrus spp) cultivars in vitro. Plant cell report, 1989(7): 688-691
[109] Cocking, E. C., Method for the isolation of plant protoplasts and vacuoles. Nature, 1960, 187-929
[110] Cupta, P. K., and Durzan, D. J. Biotechnology of somatic polyembryogenesis and plantlet regeneration in loblolly pine.Bio/Tech,1987,5:147-151
[111] Constantin, M. J., Henke R R and Wansur M A. Effect of activated carbon on callus growth and shoot organogenesis in tobacco. In vitro, 1977(13):293-296
[112] Chesick, E. E., Mohn, C. A., and Hackett, W. P., Plantlet multiplication from white pine(Pinus strobes L.)embryos in vitro: bud induction and rooting. Plant Cell Tissue Organ Culture.1991,26:107-114
[113] Campbell, R. A., and Durzan, D. J. Induction of multiple buds and needles intissue cultures of Picea glauca.Can J Bot.1975,53:1652-1657
[114] David, T., Webb, and Olga Diaz Santiago.Cytokinin induced bud formation on Caribean Pine(Pinus caribaea morlet) embryos in vitro. Plant Sci Lett .1983,32:17
[115] David, A., and David, H. Manifestations de diverses potentialities organogenes d’organes ou de fragments d’organes de pin martitime(Pinus pinaster Sol.)en culture in vitro.C R Acad Sci.Paris.1977,184:727-630
[116] David, A.，David, H., and Mateille, T. Invitro adventitious budding on Pinus pinaster otyldedons. Physiol Plant.1982,56:102-107
[117] Dunstan, D. I., Bekkaoui, F., and Pilon, M. et al. Effects of abscisic acid and analogues on the maturation of white spruce(picea glauca) somatic embryos.Plant Sci.1988,58:77
[118] Dumas, E., and Monteuuis, O. In vitro rooting of micropropagated shoots from juvenile and mature Pinus pinaster explants-influence of activated charcoal[J]. Plant Cell Tissue Org Cult, 1995(40):231-235
[119] Ebert, A., Taylor, F., and Balke, J. Changes of 6-benzylaminopurine and 2,4-dichlorphenoxyacetic acid concentrations in plant tissue culture media in the presence of activated charcoal. Plant Cell Tissue Org Cult, 1993(33):157-162
[120] Ellis, D., and Bilderback, D.E.Temporal competence of embryonic Pinus ponderosa cotyledons to form multiple buds in vitro.Amer J Bot .1989,76:348-355
[121] Ellis, D., and Bilderback, D. E. Multiple bud formation by cultured embryos of Pinus ponderosa. Plant Physiol.1984,115:201-204
[122] Ellis, D. et al. Plant Cell, Tissue and Organ Culture.1987,11(1):57
[123] Emily, E., Chesick, Carl, A., Mohn, Wesley P. Hackett. Plantet multiplication from white pine(Pinus strobus L.)embryos in vitro: bud induction and rooting.Plant Cell TISSUE and Organ Culture,1991, 26:107-114
[124] Finer, J. J., Kriebel, H. B., and Becwar, M. R. Initiation of embryogenic callus and susension cultures of eastern white pine(Pinus strobes L.). Plant Cell Rep .1989,8:203-206
[125] Farnum, P., Timmis, R., and Kulp, J. L. Biotechnology of forest yield. Science. 1983, 219:694-702
[126] Fowke, L. C., and Attree, S. M. Applied and basic studies of somatic embryogenesis in white spruce(Picea glauca) and black spruce(Picea mariana).In Soh W Y, Liu J R, Kamamine A(eds). Advances in Developmental Biology and Biotechnology of Higher Plants. Publishedby the Korean
Soc Plant Tissue Culture. Korea, 1993,5
[127] Fonda, R. W., Belanger, L. A., and Burley, L. L. Burning characteristics of western conifer needles. Northwest Sci, 1998,72: 1-9
[128] Go, N. E., Glorina, D., Perez-Orozco and Halos, S. C. In vitro response of embryos from different provenances of pinuns caribbaea var. hondurensis Morelet. Plant-Cell, Tissue and Organ Culture.1993,32:1-7
[129] Gupta, P. K., Durzan, D. J. Somatic plyembryogenesis from callus of mature sugar pine embryos. Bio/Tech.1986,4:643-645
[130] Gupta, P. K., and Durzan, D. J. Shoot multiplication from mature trees of Douglas-fir (Pseudotsuga menziesii) and sugar pine (Pinus lambertiana).Plant Cell Rep.1985,4:177-179
[131] Gupta, P. K., and Durzan, D. J. Somatic plyembryogenesis from mature trees of Douglasfir and sugar pine. Plant Cell Rep.1986,4:643-645
[132] Gladfelter, H. J., and Phillips, G. C. Denovo shoot organogenesis of Pinus eldarica in vitro. Plant Cell Rep.1987,6:163-166
[133] Gupta, P. K., and Pullman, G. S. Method for reproducing coniferous plants by somatic embryogenesis using abscisic acid and osmotic potential variation. U. S. Patent No.5,036,007, 1991
[134] Gupta, P. K., and Pullman, G. S. High concentration enrichment of conifer embryonal cells. U. S. patent No.5,041,382,1991
[135] Genys, J. B., and Hegestad, H. E. Susceptibility of different species,clones and strains of pines to acute injury caused by ozone and sulfer dioxide. Plant disease reporter,1978,6(2):687-691
[136] Halhouli, K. A,, Darwish, N. A., and Alldoon, N. M. Effects of pH and inorganic salts on the adsorption of phenol from aqueous systems on activated decoloring charcoal. Separ Sci Technol, 1995(30):3313-3324
[137] Haissig, B. E., Nelson, N. D., and Kidd, G. H. Trends in the use of tissue culture in forest improvement. Bio/Technology. 1987,5:5
[138] Haddock, P. G. Sapling sugar pine grown from excised mature embryos. Jour Forestry. 1954 , 52 : 434-437
[139] Hohtola, A. Seasonal changes in explant viability and contamination of tissue cultures from mature Scots pine Plant-Cell, Tissue and Organ Culture .1988,15:211-222
[140] James, D. J., Passey, A. J., and Rugini, E. Factors affecting high frequency plant regeneration from apple leaf tissue cultured in vitro. Journal of plant physiology, 1989(132): 148-154
[141] Jones, O. P., Effect of benzylandenine on isolated apple shoots. Nature, 1967, 215: 1514-1515
[142] Johansson, L., and Eriksson, T. Induced embryo formation in anther culture of sever anemone species. Physiol Plant, 1977(40):172-174
[143] Johansson, L., Calleberg, E., and Gedin, A. Correlation between activated charcoal, Fe-EDTA and other
organic media ingredients in cultures of anothers of Anemone canadenis. Physiol Plant, 1990(80):243-249
[144] Jiaswal, V. S., and Armin, M. N. In vitro propagation of guava from shoot cultures of mature trees. J Plant Physiol, 1987(130):7-12
[145] Jang, J. C., and Tainter, F. H. Micropropagation of shortleaf, Virginia and loblolly × shortleaf pine hybrids via organogenesis. Plant-Cell, Tissue and Organ Culture .1991,25: 61-67
[146] Jain, S. N., Dong, N., and Newton, R. J. Somatic embrygenesis in slash pine(Pinus elliottii)from immature embryo culture In vitro. Plant Sci.1989,65:233-241
[147] Kaul, K. Plant regeneration from cotyledon-hypocotyl explants of Pinus strobes L. Plant Cell Rep.1987,6:5-7
[148] Kolevska, Pletikapic, Jelaska, S., and Berllak, J. Bud and shoot formation in juvenile tissue culture of pinus nigra. Silvae genetica . 1983 , 32 : 115-119
[149] Lee, K. C., and Ahn, K. Y, A. Studying on the diversifying of plant species for the landscape construction in Korea. Seoul national university journal of agricultural sciences, 1991, 16(1): 67-80
[150] Lin, Y. Q., Wager, M. R., and Heidmann, L. J. In vitro formation of axillary buds by immature shoots of Pinus ponderosa. Plant-Cell, Tissue and Organ Culture.1991,26(3):161-166
[151] Main, M. L., Marin, J. A. Excised roots cultured in vitro. Plant Cell Report. 1998,18(3/4):350
[152] Merkle, S. A. Plant Tissue Culture Concept and laboratory exercise.CRC Press,1996
[153] M’Kada, J., Dorin, N., and Bigot, C. In vitro micropropagation of Arctostuphylos uvaursi L. Sperengel: comparison between two methodologies.Plant Cell Tissue Org Cult, 1991(24):217-222
[154] Mehra, Palta, A., Smeltzer, R. H., and Mott, R. L. Hormonal control of induced organogenesis : Experiments with excised plant parts of loblolly pine.Tappi. 1978, 61:37 ~40
[155] Mott, R. L., and Amerson, H. V. Tissue culture plantlets produced from Pinus monticola embryonic materials. Forest Sci . 1981 , 27 : 299~304
[156] Minocha, S. C.Callus and adventitious shoot formation in excised embryos of white pine(Pinus strobes). Can J Bot.1980,58:366-370
[157] Mazarl, A., and Camm, E. L. Effect of cytokinins on development and photosynthetic polypeptides during organogenesis of Pinus ponderosa Dougl.cytolodons cultured in vitro.Tissue and Organ Culture.1993,33:81-89
[158] Nancy, E. G., Glorinm, D., and Perez, Orzco, et al. In vitro response of embryos form different provenances of pinus caribaec var.boudurensis Morelet. Plant Cell ,Tissue and Organ Culture.1993,32(1):47-53
[159] Proskauer, J., and Berman, R. Agar culture medium modified to approximate soil condition. Nature(London), 1970(227):1161
[161] Patel, K. R., Shekhwat, N. S., Berlyn, G. P., and Thorpe, T, A. Isolation and culture of protoplasts from
cotyledons of pinus coulteri D.Don.Plant-Cell, Tissue and Organ Culture.1984,3:85-90
[161] Patel, K. R., Kim, H. R., and Thorpe, T. A. Plantlet formation in pitch pine by tissue culture methods. Forest Ecol Managenent.1986,15:147-160
[162] Patel, K. R., and Thorpe, T. A. In vitro differention of plantlets from embryonic explants of lodgepole pine (pinuns contorta Dougl. Ex Lound. Plant-Cell, Tissue and Organ Culture.1984,3:131-142
[163] Pulido, C. M,, Harry, I. S., and Thorpe, T. A. Optimization of bud induction in cotyledonary explants of pinus canariensis. Plant-Cell, Tissue and Organ Culture.1992,29:247-255
[164] Pulido, C. M., Harry, I. S., and Thorpe, T. A. Effect of various bud induction treatments on elongation and rooting of adventitious shoots of Canary Island pine (Pinus canariensis) Plant-Cell, Tissue and Organ Culture.1994,39:225-230
[165] Perez, Bermuudez, P. and Sommer, H. E. Factor affecting adventitious and induction in Pinus elliottii Engelm embryos cultured invitro. Plant Cell, Tissue and Organ Culture. 1987,(11):25-35
[166] Pullman, G. S., and Gupta, P. K. Method for reproducing coniferous plants by somatic embryogenesis using absorbent materials in the development stage media. U.S. patent No. 5,034,326,1991
[167] Rancillac, M., Faye, M., and David, A. In vitro rooting of cloned shoots in Pinus pinaster. Physiol Plant.1982,56:97-101
[168] Robert, D. R. Abscisic acid and mannitol promote early development,maturation and storage protein accumilation in somatic embryos of interior spruce. Physiol Plant.1991,83:247
[169] Skoog, F. C. Tsui. Chemical control of growth and bud formation in tobacco stem segments and callus cultured in vitrol. Amer. J. Bot.,1948,(35):782-787
[170] Skoog, F. C. Tsui. Growth substances and the foemation of buds in plant tissue. In:Plant growth substance,1951,p265-285
[171] Steward, F.C., Holsten, J. R. D., and Simth, J. Growth and organized development of cultured cellsⅡ: growth and division of freely suspended cells. Amer J Bot. 1958,（45）:693-703
[172] Sommer, H. E., Brown, C. L., and Kormanik, P. P. Differentiation of plantlets in long-leaf pine(Pinus palustris Mill) tissue cultured in vitro. Bot Gaz.1975,136:196~200
[173] Sommer, H. E., and Brown, C. L. Plantlets formation in pine tissue cultures. Amer J Bot. Suppl. 1974,61:11
[174] Schwarz, O. J., Schlarbaum, S. E., and Beatu, R. M. Plantlet regeneration form mature zygotic embryos of eastern white pine(Pinus strobes L.). Plant Cell Rep.1988,7:174-174
[175] Takebe, T. Regulation of whole plants from isolated mesophyll protoplasts of tobacoo. Nature, 1971,58: 318-320
[176] Tranvan, H. In vitro adventitious bud formation on isolated seedlings of Pinus sylvestris L.Biologia Plantarum(Praha). 979,21(3):230-233
[177] Tuskan, G. A., Sargent, W. A., and Rensem, T. et al. Influence of plant growth regulators, basal media
and Carbohydrate levels on the in vitro development of Pinus ponderosa cotyledon explants.Plant Cell,Tissue and organ culture,1990,20(1): 47-52
[178] Teasale, R. D., Dawson, P. A, and Woolhouse, H. W. Mineral nutrient requirements of a loblolly pine(Pinus taeda) cell suspension culture,Plant Physiol.986,82: 942-945
[179] Tautorus, T. E,, Fowke, L. C., and Dunstan, D. I. Somatic embryogenes is in conifers. Can J Bot,1991,69:1873
[180] Ueda, N., Dewa, N., and Tsuneya, T., et al. The leaf oil components of lacebark pine, Pinus bungeana Zuuc.Chemistry Express,1989,4(12):813-816
[181] Von Arnold, S., and Eriksson, T. Invitro studies of adventitious shoot formation in Pinus contorta .Can J Bot.1981,59:870-874
[182] White, P. R.The cultivation of animal and plant cells.1943
[183] Whitehead, HCM, K. L. Giles Rapid propagation of polar by tissue culture method . NZ J For Sci 1977,(7):40-43
[184] Weatherhead, M. A., Burdon, J., and Henshaw, G. G. Some effects of activated charcoal as an additive to plant tissue culture media. Z pflanzen-physiol,1978(89):141-147
[185] Webb, D. T., and Santiago, O. D. Induced bud formation on Caribbean pine(Pinus caribaea Morelet)embryos in vitro. Plant Sci Lett.1983,32:17-21
[186] Yam, T. Y., Erns, R., and Arditi, J., et al. Charcoal in orchid seed germination and tissue culture media: a review. Lindleyana,1990(5):256-165