Establishment of Cell Suspension Culture and Growth Characteristics of Wild Nitraria tangutorum

NI Jian-wei; WEI Qi; YANG Xiu-yan; XU Xiu-yu; WU Hai-wen; ZHANG Hua-xin; LIU Tao

Volume 28 Issue 1

Article Contents

Turn off MathJax

Article Navigation > Forest Research > 2015 > 28(1): 74-80

Citation:

Establishment of Cell Suspension Culture and Growth Characteristics of Wild Nitraria tangutorum

1.
Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, China
2.
Research Center of Saline and Alkali Land of State Forestry Administration, Beijing 100091, China
3.
Guangdong Academy of Forestry, Guangzhou 510520, Guangdong, China

Received Date: 2014-08-26

Abstract

Taking stems and leaves of wild Nitraria tangutorum Bobr. as explants to study the disinfectant effects by NaClO on explants and the effects of 2,4-D, NAA and 6-BA on callus induction and morphology. The callus morphology was analyzed and cell suspension systems were established with the loose type callus, its growth characteristics were observed. The results show that the best way of disinfection of explants was 10 minutes with NaClO, the longer or shorter the sterilization time, the lower the survival rate. Three hormones had effects on relative growth and morphology of callus and the 2,4-D was a major factor, the relative growth rate of callus from stem was higher than that of leaves. To be confirmed by callus growth rate and morphological characteristics, cultured on MS medium containing 2,4-D 1.5 mg·L-1, NAA 0.2 mg·L-1 and 6-BA 0.4 mg·L-1 was the optimum treatment combination for the establishment of N. tangutorum suspension cell lines. The calluses were loose type with light yellow, and the best inoculation dosage was 7.5 mL mother liquor. Under such condition, the cells grew in an 'S' curve. The cell mitotic index reached the maximum value of 5.1% at the seventh day, at the third day of cultured the cell got the strongest activity and the light absorption value was 0.69. The survival rate in the lag phase and stationary phase decreased slightly faster than logarithmic phase, but maintained between 84%-93%.
- Nitraria tangutorum.,
- callus,
- suspension cells,
- cell culture,
- growth characteristics

References

[1]	李双福,张启昌,张起超,等. 白刺属植物研究进展[J]. 北华大学学报:自然科学版,2005,6(1):78-81.
[2]	潘晓玲,沈观冕,陈鹏. 白刺属植物的分类学及系统学研究[J]. 云南植物研究,1999,21(3):21-29.
[3]	潘晓云,曹琴东,尉秋实,等. 白刺属的系统进化和生物多样性研究进展[J]. 中国医学生物技术应用,2002(4):1-6.
[4]	Duan J, Williams I D, Che C, et al. Tangutorine: A novel β-carboline alkaloid from Nitraria tangutorum[J]. Tetrahedron letters,1999, 40(13):2593-2596.
[5]	宋永朋,张东,林辉. 白刺果皮中原花青素的提取工艺[J]. 食品研究与开发,2012,33(8):85-88.
[6]	任旭,陈贵林. 唐古特白刺果实提取物抗氧化活性评价[J]. 食品科学,2011,32(3):95-97.
[7]	高海宁,李彩霞,张勇,等. 唐古特白刺果浆中原花青素提取工艺及抗氧化性研究[J]. 食品工业科技,2010,31(10):109-112.
[8]	刘利平,燕玲. 白刺属植物化学成分的研究现状与发展趋势[J]. 内蒙古农业大学学报:自然科学版,2009,30(1):316-320.
[9]	马启慧. 耐盐碱植物白刺的开发与利用[J]. 黑龙江农业科学,2007(5):116-117.
[10]	何正伦. 白刺的离体繁殖[J]. 甘肃林业科技,1989 (2):52-53.
[11]	王尚德. 唐古特白刺优株选择与组织培养研究 [D]. 北京:北京林业大学, 2005.
[12]	张红晓,康向阳. 白刺组织培养技术的研究[J]. 西北植物学报,2004,24(1):56-64.
[13]	李代丽. 白刺愈伤组织培养中外源激素对内源激素影响的研究 [D]. 北京:北京林业大学,2007.
[14]	Yang Y, Wei X, Shi R, et al. Salinity-induced Physiological Modification in the Callus from Halophyte Nitraria tangutorum Bobr.[J]. Journal of plant growth regulation,2010,29(4):465-476.
[15]	Zhang C H, Mei X G, Liu L, et al. Enhanced paclitaxel production induced by the combination of elicitors in cell suspension cultures of Taxus chinensis[J]. Biotechnology Letters,2000,22(19):1561-1564.
[16]	Carrier D, Chauret N, Mancini M, et al. Detection of ginkgolide A in Ginkgo biloba cell cultures[J]. Plant cell reports,1991,10(5):256-259.
[17]	Lu M, Wong H, Teng W. Effects of elicitation on the production of saponin in cell culture of Panax ginseng[J]. Plant Cell Reports,2001,20(7):674-677.
[18]	杜晓映,张振文,夏惠,等. 葡萄悬浮细胞系的建立[J]. 西北农林科技大学学报:自然科学版,2009, 37(1):199-210.
[19]	Guo Z, Liu Y, Gong M, et al. Regulation of vinblastine biosynthesis in cell suspension cultures of catharanthus roseus[J]. Plant Cell, Tissue and Organ Culture,2013,112(1):43-54.
[20]	Namdeo A G. Plant cell elicitation for production of secondary metabolites: a review[J]. Pharmacognosy reviews,2007,1(1):69-79.
[21]	Towill L E, Mazur P. Studies on the reduction of 2, 3, 5-triphenyltetrazolium chloride as a viability assay for plant tissue cultures[J]. Canadian Journal of Botany,1975, 53(11):1097-1102.
[22]	王晨霞. 西伯利亚白刺离体培养体系建立及试管苗耐盐性研究 [D]. 内蒙古呼和浩特:内蒙古大学,2008.
[23]	范小峰,杨颖丽,刘秀丽,等. 唐古特白刺愈伤组织诱导及再生体系的研究[J]. 西北师范大学学报:自然科学版,2009,45(2):74-77.
[24]	王海波,魏景芳,葛亚新,等. 小麦愈伤组织状态调控与原生质体培养[J]. 中国农业科学,1996,29(6): 8-14.
[25]	孙敬三,朱至清. 植物细胞工程实验技术[M]. 北京:化学工业出版社,2006.
[26]	刘磊,顾玉红,孟坤,等. 文冠果悬浮细胞系的建立及细胞生长特性[J]. 林业科学,2010,46(9):79-83.
[27]	闫静辉,张小兵,李亚璞,等. 西洋参悬浮细胞系的建立及其生长特性的研究[J]. 河北省科学院学报,2005,22(4):23-26,36.
[28]	朱玉贤,李心毅. 现代分子生物学[M]. 北京:高等教育出版社,2002.
[29]	李勇. 桑树细胞悬浮培养的研究 [D]. 山东泰安:山东农业大学,2007.
[30]	胡风庆,任娟. 东北红豆杉细胞悬浮培养研究[J]. 辽宁大学学报:自然科学版,2002,29(3):279-282.

Proportional views

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Get Citation

PDF

XML

Article views(3005) PDF downloads(1012) Cited by()

Proportional views

Establishment of Cell Suspension Culture and Growth Characteristics of Wild Nitraria tangutorum

1. Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, China

2. Research Center of Saline and Alkali Land of State Forestry Administration, Beijing 100091, China

3. Guangdong Academy of Forestry, Guangzhou 510520, Guangdong, China

Received Date: 2014-08-26

Abstract: Taking stems and leaves of wild Nitraria tangutorum Bobr. as explants to study the disinfectant effects by NaClO on explants and the effects of 2,4-D, NAA and 6-BA on callus induction and morphology. The callus morphology was analyzed and cell suspension systems were established with the loose type callus, its growth characteristics were observed. The results show that the best way of disinfection of explants was 10 minutes with NaClO, the longer or shorter the sterilization time, the lower the survival rate. Three hormones had effects on relative growth and morphology of callus and the 2,4-D was a major factor, the relative growth rate of callus from stem was higher than that of leaves. To be confirmed by callus growth rate and morphological characteristics, cultured on MS medium containing 2,4-D 1.5 mg·L-1, NAA 0.2 mg·L-1 and 6-BA 0.4 mg·L-1 was the optimum treatment combination for the establishment of N. tangutorum suspension cell lines. The calluses were loose type with light yellow, and the best inoculation dosage was 7.5 mL mother liquor. Under such condition, the cells grew in an 'S' curve. The cell mitotic index reached the maximum value of 5.1% at the seventh day, at the third day of cultured the cell got the strongest activity and the light absorption value was 0.69. The survival rate in the lag phase and stationary phase decreased slightly faster than logarithmic phase, but maintained between 84%-93%.

HTML

Reference (30)

[1]	李双福,张启昌,张起超,等. 白刺属植物研究进展[J]. 北华大学学报:自然科学版,2005,6(1):78-81.
[2]	潘晓玲,沈观冕,陈鹏. 白刺属植物的分类学及系统学研究[J]. 云南植物研究,1999,21(3):21-29.
[3]	潘晓云,曹琴东,尉秋实,等. 白刺属的系统进化和生物多样性研究进展[J]. 中国医学生物技术应用,2002(4):1-6.
[4]	Duan J, Williams I D, Che C, et al. Tangutorine: A novel β-carboline alkaloid from Nitraria tangutorum[J]. Tetrahedron letters,1999, 40(13):2593-2596.
[5]	宋永朋,张东,林辉. 白刺果皮中原花青素的提取工艺[J]. 食品研究与开发,2012,33(8):85-88.
[6]	任旭,陈贵林. 唐古特白刺果实提取物抗氧化活性评价[J]. 食品科学,2011,32(3):95-97.
[7]	高海宁,李彩霞,张勇,等. 唐古特白刺果浆中原花青素提取工艺及抗氧化性研究[J]. 食品工业科技,2010,31(10):109-112.
[8]	刘利平,燕玲. 白刺属植物化学成分的研究现状与发展趋势[J]. 内蒙古农业大学学报:自然科学版,2009,30(1):316-320.
[9]	马启慧. 耐盐碱植物白刺的开发与利用[J]. 黑龙江农业科学,2007(5):116-117.
[10]	何正伦. 白刺的离体繁殖[J]. 甘肃林业科技,1989 (2):52-53.
[11]	王尚德. 唐古特白刺优株选择与组织培养研究 [D]. 北京:北京林业大学, 2005.
[12]	张红晓,康向阳. 白刺组织培养技术的研究[J]. 西北植物学报,2004,24(1):56-64.
[13]	李代丽. 白刺愈伤组织培养中外源激素对内源激素影响的研究 [D]. 北京:北京林业大学,2007.
[14]	Yang Y, Wei X, Shi R, et al. Salinity-induced Physiological Modification in the Callus from Halophyte Nitraria tangutorum Bobr.[J]. Journal of plant growth regulation,2010,29(4):465-476.
[15]	Zhang C H, Mei X G, Liu L, et al. Enhanced paclitaxel production induced by the combination of elicitors in cell suspension cultures of Taxus chinensis[J]. Biotechnology Letters,2000,22(19):1561-1564.
[16]	Carrier D, Chauret N, Mancini M, et al. Detection of ginkgolide A in Ginkgo biloba cell cultures[J]. Plant cell reports,1991,10(5):256-259.
[17]	Lu M, Wong H, Teng W. Effects of elicitation on the production of saponin in cell culture of Panax ginseng[J]. Plant Cell Reports,2001,20(7):674-677.
[18]	杜晓映,张振文,夏惠,等. 葡萄悬浮细胞系的建立[J]. 西北农林科技大学学报:自然科学版,2009, 37(1):199-210.
[19]	Guo Z, Liu Y, Gong M, et al. Regulation of vinblastine biosynthesis in cell suspension cultures of catharanthus roseus[J]. Plant Cell, Tissue and Organ Culture,2013,112(1):43-54.
[20]	Namdeo A G. Plant cell elicitation for production of secondary metabolites: a review[J]. Pharmacognosy reviews,2007,1(1):69-79.
[21]	Towill L E, Mazur P. Studies on the reduction of 2, 3, 5-triphenyltetrazolium chloride as a viability assay for plant tissue cultures[J]. Canadian Journal of Botany,1975, 53(11):1097-1102.
[22]	王晨霞. 西伯利亚白刺离体培养体系建立及试管苗耐盐性研究 [D]. 内蒙古呼和浩特:内蒙古大学,2008.
[23]	范小峰,杨颖丽,刘秀丽,等. 唐古特白刺愈伤组织诱导及再生体系的研究[J]. 西北师范大学学报:自然科学版,2009,45(2):74-77.
[24]	王海波,魏景芳,葛亚新,等. 小麦愈伤组织状态调控与原生质体培养[J]. 中国农业科学,1996,29(6): 8-14.
[25]	孙敬三,朱至清. 植物细胞工程实验技术[M]. 北京:化学工业出版社,2006.
[26]	刘磊,顾玉红,孟坤,等. 文冠果悬浮细胞系的建立及细胞生长特性[J]. 林业科学,2010,46(9):79-83.
[27]	闫静辉,张小兵,李亚璞,等. 西洋参悬浮细胞系的建立及其生长特性的研究[J]. 河北省科学院学报,2005,22(4):23-26,36.
[28]	朱玉贤,李心毅. 现代分子生物学[M]. 北京:高等教育出版社,2002.
[29]	李勇. 桑树细胞悬浮培养的研究 [D]. 山东泰安:山东农业大学,2007.
[30]	胡风庆,任娟. 东北红豆杉细胞悬浮培养研究[J]. 辽宁大学学报:自然科学版,2002,29(3):279-282.

Establishment of Cell Suspension Culture and Growth Characteristics of Wild Nitraria tangutorum

Abstract

References

Proportional views

通讯作者: 陈斌, bchen63@163.com

Proportional views

Establishment of Cell Suspension Culture and Growth Characteristics of Wild Nitraria tangutorum

HTML

Catalog

Export File

Citation

Format

Content