Establishment of an Efficient Regeneration System of Populus×euramericana cl. 'Bofeng 1’

CUI Xu-dong; SU Xiao-hua; ZHANG Bing-yu; CHU Yan-guang; HUANG Qin-jun; ZHANG Wei-xi; LIU Bo-yang

Volume 25 Issue 2

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Establishment of an Efficient Regeneration System of Populus×euramericana cl. 'Bofeng 1’

1.
State Key Laboratory of Forest Genetics and Tree Breeding, Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, China
2.
Key Laboratory of Genetics and Breeding in Forestry Trees Ornamental Plants, Ministry of Education, College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083, China

Received Date: 2011-12-28

Abstract

Compared with the species of Leuce section, the difficulty in plant regeneration during tissue culture for the species of Aigeiros section is a key limiting factor for their application in genetic engineering of forest trees. In this study, the plant growth regulators combination, copper concentration, light intensity, and the selection pressures of kanamycin were investigated using leaves of Populus×euramericana cl. 'Bofeng 1’ as explants, and a stable and high efficient regeneration system was established. Using this system, both the shoot regeneration rate and rooting rate were up to 100%, and the average number of differentiated shoots in each leaf explant was up to 20, and the survival rate of seedlings reached 98%. The optimal selection pressure of kanamycin in shoot inducement for leaf-explant was 40 mg·L-1, and the optimal selection pressure of kanamycin in root inducement of adventitious bud was 20 mg·L-1. The authors also found that differentiation rate of adventitious buds of Populus×euramericana cl. 'Bofeng 1’ could be significantly promoted by increased copper concentration, and this is the first report about the key role of copper (Cu) in explant regeneration of Poplars. This regeneration system with high frequency which was established by the authors was as good as that of species of Leuce section, and it provides a good technological platform for genetic engineering of forest tree which uses the species of Aigeiros section as model materials.
- Populus×euramericana cl. 'Bofeng 1’,
- high frequency regeneration system,
- copper

References

[1]	Winton L. Tissue Culture Propagation of European Aspen[J]. Forest Science, 1971, 17(3): 348-350
[2]	Ahuja M R. Somatic cell differentiation and rapid clonal propagation of aspen [J]. Silvae Genet, 1983, 32(3-4): 131-135
[3]	Coleman G D, Ernst S G. In vitro shoot regeneration of Populus deltoides: effect of cytokinin and genotype[J]. Plant Cell Pep, 1989, 8(8): 459-462
[4]	Vinocur B, Carmi T, Altman A, et al. Enhanced bud regeneration in aspen (Populus tremula L.) roots cultured in liquid media[J]. Plant Cell Rep, 2000, 19(12): 1146-1154
[5]	Wang H M, Liu H M,Wang W J, et al. Effects of Thidiazuron, basal medium and light quality on adventitious shoot regeneration from in vitro cultured stem of Populus alba ×P. berolinensis[J]. Journal of Forestry Research, 2008, 19(3): 257-259
[6]	Bosela M J. Effects of β-lactam antibiotics, auxins, and cytokinins on shoot regeneration from callus cultures of two hybrid aspens, Populus tremuloides×P. tremula and P. xcanescens×P. gradidentata[J]. Plant cell, tissue and organ culture, 2009, 98(3): 249-261
[7]	Yevtushenko D P, Misra S. Efficient Agrobacterium-mediated transformation of commercial hybrid poplar Populus nigra L.×P. maximowiczii A. Henry[J]. Plant Cell Rep, 2010, 29(3): 211–221
[8]	陈彩霞,沈昕,王祎宁,等. 派间杂种110杨再生系统的建立[J]. 林业科学研究,2007,20(2):287-291
[9]	张蕾,苏晓华,张冰玉,等. 京2杨组培条件的优化及再生体系建立[J]. 林业科学研究,2007,20(6):787-793
[10]	Gary D C, Stephen G E. In vitro shoot regeneration of Populus deltoides: effect of cytokinin and genotype[J]. Plant Cell Rep, 1989, 8(8): 459–462
[11]	Chaturvedi H C, Sharma A K, Agha B Q, et al. Production of cloned trees of Populus deltoides through in vitro regeneration of shoots from leaf, stem and root explants and their field cultivation[J]. Indian Journal of Biotechnology, 2004, 3(2): 203-208
[12]	Mingozzi M, Montello P, Merkle S. Adventitious shoot regeneration from leaf explants of eastern cottonwood (Populus deltoides) cultured under photoautotrophic conditions[J].Tree Physiology, 2009, 29(3): 333-343
[13]	苏晓华,张冰玉,黄秦军,等. 我国林木基因工程研究进展及关键领域[J]. 林业科学,2003,39(5):111-118
[14]	苏晓华,张冰玉,黄秦军,等. 杨树基因工程育种[M]. 北京:科学出版社,2009
[15]	马常耕. 我国杨树育种中的若干问题商榷[J]. 青海农林科技,2004(B03):1-8
[16]	苏晓华,丁昌俊,马常耕. 我国杨树育种的研究进展及对策[J]. 林业科学研究,2010, 23(1): 31-37
[17]	陈正华. 木本植物组织培养及其应用[M]. 北京:高等教育出版社,1986
[18]	王斌,李百炼,张金凤,等. 杂种白杨离体再生体系的建立[J]. 西北植物学报,2009,29(4):704-710
[19]	王红蕾. 山新杨高效组培再生体系的建立[J]. 北方园艺,2010(13):174-175
[20]	Purnhauser L. Stimulation of shoot and root regeneration in wheat, Triticum aestivum callus cultures by copper[J]. Cereal Res. Commun, 1991, 19: 419- 423
[21]	杨跃生,筒玉瑜,郑迎. 铜在水稻愈伤组织培养再生植株中的促进作用[J]. 中国水稻科学,1999,13(2):95-98
[22]	李会勇,尹钧,刘雷. Cu²⁺浓度对啤酒大麦幼胚组织培养与植株再生的影响[J]. 麦类作物学报,2003,23(2):27-29
[23]	王树昌,于晓玲,赵平娟. Cu离子对巴西橡胶树组织培养的影响[J]. 热带农业工程,2010,34(6):28-30
[24]	Purnhauser L, Gyulai. Effect of copper on shoot and root regeneration in wheat, triticale, rape and tobacco tissue culture[J]. Plant Cell, Tissue and Organ Culture, 1993, 35: 131-139
[25]	Jain P, Kachhwaha S, Kothari S L. Improved micropropagation protocol and enhancement in biomass and chlorophyll content in Stevia rebaudiana (Bert.) Bertoni by using high copper levels in the culture medium[J]. Scientia Horticulturae, 2009, 119(3): 315-319

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通讯作者: 陈斌, bchen63@163.com

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沈阳化工大学材料科学与工程学院沈阳 110142

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Establishment of an Efficient Regeneration System of Populus×euramericana cl. 'Bofeng 1’

1. State Key Laboratory of Forest Genetics and Tree Breeding, Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, China

2. Key Laboratory of Genetics and Breeding in Forestry Trees Ornamental Plants, Ministry of Education, College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083, China

Received Date: 2011-12-28

Available Online: 2012-04-13

Abstract: Compared with the species of Leuce section, the difficulty in plant regeneration during tissue culture for the species of Aigeiros section is a key limiting factor for their application in genetic engineering of forest trees. In this study, the plant growth regulators combination, copper concentration, light intensity, and the selection pressures of kanamycin were investigated using leaves of Populus×euramericana cl. 'Bofeng 1’ as explants, and a stable and high efficient regeneration system was established. Using this system, both the shoot regeneration rate and rooting rate were up to 100%, and the average number of differentiated shoots in each leaf explant was up to 20, and the survival rate of seedlings reached 98%. The optimal selection pressure of kanamycin in shoot inducement for leaf-explant was 40 mg·L-1, and the optimal selection pressure of kanamycin in root inducement of adventitious bud was 20 mg·L-1. The authors also found that differentiation rate of adventitious buds of Populus×euramericana cl. 'Bofeng 1’ could be significantly promoted by increased copper concentration, and this is the first report about the key role of copper (Cu) in explant regeneration of Poplars. This regeneration system with high frequency which was established by the authors was as good as that of species of Leuce section, and it provides a good technological platform for genetic engineering of forest tree which uses the species of Aigeiros section as model materials.

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[1]	Winton L. Tissue Culture Propagation of European Aspen[J]. Forest Science, 1971, 17(3): 348-350
[2]	Ahuja M R. Somatic cell differentiation and rapid clonal propagation of aspen [J]. Silvae Genet, 1983, 32(3-4): 131-135
[3]	Coleman G D, Ernst S G. In vitro shoot regeneration of Populus deltoides: effect of cytokinin and genotype[J]. Plant Cell Pep, 1989, 8(8): 459-462
[4]	Vinocur B, Carmi T, Altman A, et al. Enhanced bud regeneration in aspen (Populus tremula L.) roots cultured in liquid media[J]. Plant Cell Rep, 2000, 19(12): 1146-1154
[5]	Wang H M, Liu H M,Wang W J, et al. Effects of Thidiazuron, basal medium and light quality on adventitious shoot regeneration from in vitro cultured stem of Populus alba ×P. berolinensis[J]. Journal of Forestry Research, 2008, 19(3): 257-259
[6]	Bosela M J. Effects of β-lactam antibiotics, auxins, and cytokinins on shoot regeneration from callus cultures of two hybrid aspens, Populus tremuloides×P. tremula and P. xcanescens×P. gradidentata[J]. Plant cell, tissue and organ culture, 2009, 98(3): 249-261
[7]	Yevtushenko D P, Misra S. Efficient Agrobacterium-mediated transformation of commercial hybrid poplar Populus nigra L.×P. maximowiczii A. Henry[J]. Plant Cell Rep, 2010, 29(3): 211–221
[8]	陈彩霞,沈昕,王祎宁,等. 派间杂种110杨再生系统的建立[J]. 林业科学研究,2007,20(2):287-291
[9]	张蕾,苏晓华,张冰玉,等. 京2杨组培条件的优化及再生体系建立[J]. 林业科学研究,2007,20(6):787-793
[10]	Gary D C, Stephen G E. In vitro shoot regeneration of Populus deltoides: effect of cytokinin and genotype[J]. Plant Cell Rep, 1989, 8(8): 459–462
[11]	Chaturvedi H C, Sharma A K, Agha B Q, et al. Production of cloned trees of Populus deltoides through in vitro regeneration of shoots from leaf, stem and root explants and their field cultivation[J]. Indian Journal of Biotechnology, 2004, 3(2): 203-208
[12]	Mingozzi M, Montello P, Merkle S. Adventitious shoot regeneration from leaf explants of eastern cottonwood (Populus deltoides) cultured under photoautotrophic conditions[J].Tree Physiology, 2009, 29(3): 333-343
[13]	苏晓华,张冰玉,黄秦军,等. 我国林木基因工程研究进展及关键领域[J]. 林业科学,2003,39(5):111-118
[14]	苏晓华,张冰玉,黄秦军,等. 杨树基因工程育种[M]. 北京:科学出版社,2009
[15]	马常耕. 我国杨树育种中的若干问题商榷[J]. 青海农林科技,2004(B03):1-8
[16]	苏晓华,丁昌俊,马常耕. 我国杨树育种的研究进展及对策[J]. 林业科学研究,2010, 23(1): 31-37
[17]	陈正华. 木本植物组织培养及其应用[M]. 北京:高等教育出版社,1986
[18]	王斌,李百炼,张金凤,等. 杂种白杨离体再生体系的建立[J]. 西北植物学报,2009,29(4):704-710
[19]	王红蕾. 山新杨高效组培再生体系的建立[J]. 北方园艺,2010(13):174-175
[20]	Purnhauser L. Stimulation of shoot and root regeneration in wheat, Triticum aestivum callus cultures by copper[J]. Cereal Res. Commun, 1991, 19: 419- 423
[21]	杨跃生,筒玉瑜,郑迎. 铜在水稻愈伤组织培养再生植株中的促进作用[J]. 中国水稻科学,1999,13(2):95-98
[22]	李会勇,尹钧,刘雷. Cu²⁺浓度对啤酒大麦幼胚组织培养与植株再生的影响[J]. 麦类作物学报,2003,23(2):27-29
[23]	王树昌,于晓玲,赵平娟. Cu离子对巴西橡胶树组织培养的影响[J]. 热带农业工程,2010,34(6):28-30
[24]	Purnhauser L, Gyulai. Effect of copper on shoot and root regeneration in wheat, triticale, rape and tobacco tissue culture[J]. Plant Cell, Tissue and Organ Culture, 1993, 35: 131-139
[25]	Jain P, Kachhwaha S, Kothari S L. Improved micropropagation protocol and enhancement in biomass and chlorophyll content in Stevia rebaudiana (Bert.) Bertoni by using high copper levels in the culture medium[J]. Scientia Horticulturae, 2009, 119(3): 315-319

Establishment of an Efficient Regeneration System of Populus×euramericana cl. 'Bofeng 1’

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通讯作者: 陈斌, bchen63@163.com

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Establishment of an Efficient Regeneration System of Populus×euramericana cl. 'Bofeng 1’

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