Investigation of browning factors in embryogenic callus culture of <i>Pinus tabulaeformis</i> and optimization of proliferation medium

FU Shuangbin; LI Hui; ZHAO Jian; FU Xuejun; ZHU Songlin; ZHANG Jinfeng

doi:10.7671/j.issn.1001-411X.2017.05.016

Journal of South China Agricultural University > 2017 > 38(5): 91-96. > DOI: 10.7671/j.issn.1001-411X.2017.05.016

FU Shuangbin, LI Hui, ZHAO Jian, FU Xuejun, ZHU Songlin, ZHANG Jinfeng. Investigation of browning factors in embryogenic callus culture of Pinus tabulaeformis and optimization of proliferation medium[J]. Journal of South China Agricultural University, 2017, 38(5): 91-96. DOI: 10.7671/j.issn.1001-411X.2017.05.016

Citation:

PDF (1772 KB)

Investigation of browning factors in embryogenic callus culture of Pinus tabulaeformis and optimization of proliferation medium

1.
National Engineering Laboratory for Tree Breeding/Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants of Ministry of Education/Key Laboratory of Forest Trees and Ornamental Plants Biological Engineering of State Forestry Administration/College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China
2.
National Pinus tabulaeformis seed base, State-owned Forest Administration of Lüliang Mountain, Linfen 041300, China

More Information

Received Date: November 30, 2016
Available Online: May 17, 2023

Graphical Abstract

Abstract

Abstract

Objective
To study the effects of different factors on browning of embryogenic callus of Pinus tabulaeformis during proliferation, and optimize the culture medium based on proliferation rate for lower browning rate and higher efficiency.

Method
Embryogenic callus of P. tabulaeformis from nine cell lines and three different year were used as material.We performed a serious of single factor experiments and calculated the browning rates and proliferation rates of embryogenic callus under the treatments of different callus number, pH, phytagel content, type and content of sugars, and culturing time. Orthogonal experiment was then conducted to screen out the best culture conditions.

Result
Results of the single factor experiments showed that browning rate was significantly different among different genotypes, and it was significantly positively correlated with the age of subculture. The browning rate was relatively low and the proliferation rate was relatively high when the callus number in each petri dish was 6–7, medium pH was 5.8–6.0, phytagel was 2.0–3.0 g·L^–1, and sucrose was 10 g·L^–1. Sucrose was better than glucose and maltose.

Conclusion
Based on the orthogonal design, placing 7 callus in each 90 mm petri dish, adding 2.5 g·L^–1 phytagel and 10 g·L^–1 sucrose to the medium, and adjusting the pH to 5.9 could achieve the preferred browning rate, proliferation rate and efficiency.
- Pinus tabulaeformis,
- embryogenic callus,
- browning,
- proliferation,
- lorthogonal design

FullText(HTML)

References (20)

References

[1]	沈熙环. 油松、华北落叶松良种选育实践与理论[M]. 北京: 科学出版社, 2013.
[2]	翟晓巧. 木本植物组织培养褐化控制策略[J]. 河南林业科技, 2008, 28(1): 38-40.
[3]	罗丽华. 板栗组织培养及褐变研究[D]. 长沙: 中南林学院, 2004, 9-17.
[4]	邹英宁, 吴强盛. 果树组织培养中褐变现象及其抑制研究进展[J]. 长江大学学报(自科版), 2007, 4(3): 47-50.
[5]	庞勇. 果树组织培养中褐化现象的研究进展[J]. 甘肃林业科技, 2004, 29(1): 16-18.
[6]	PIRTTILA A M, JOENSUU P, POSPIECH H, et al. Bud endophytes of Scots pine produce adenine derivatives and other compounds that affect morphology and mitigate browning of callus cultures[J]. Physiol Plant, 2004, 121(2): 305-312.
[7]	KHOSROUSHAHI A Y, NADERI-MANESH H, SIMONSEN H T. Effect of antioxidants and carbohydrates in callus cultures of Taxus brevifolia: Evaluation of browning, callus growth, total phenolics and paclitaxel production[J]. Bioimpacts, 2011, 1(1): 37-45.
[8]	陈建勇. 杉木愈伤组织培养中的褐化控制研究[J]. 亚热带植物科学, 2011, 40(3): 47-49.
[9]	BUSINGE E, EGERTSDOTTER U. A possible biochemical basis for fructose-induced inhibition of embryo development in Norway spruce (Picea abies)[J]. Tree Physiol, 2014, 34(6): 657-669.
[10]	LITVAY J D, VERMA D C, JOHNSON M A. Influence of a loblolly pine (Pinus taeda L.) culture medium and its components on growth and somatic embryogenesis of the wild carrot (Daucus carota L.)[J]. Plant Cell Rep, 1985, 4(6): 325-328.
[11]	BRETON D, HARVENGT L, TRONTIN J F, et al. High subculture frequency, maltose-based and hormone-free medium sustained early development of somatic embryos in maritime pine[J]. In Vitro Cell Dev Biol Plant, 2005, 41(4): 494-504.
[12]	KONG L, VON ADERKAS P. A novel method of cryopreservation without a cryoprotectant for immature somatic embryos of conifer[J]. Plant Cell Tiss Organ Cult, 2011, 106(1): 115-125.
[13]	SILVEIRA V, FLOH E S, HANDRO W, et al. Effect of plant growth regulators on the cellular growth and levels of intracellular proteins, starch and polyamines in embryogenic suspension cultures of Pinus taeda[J]. Plant Cell Tiss Organ Cult, 2004, 76(1): 53-60.
[14]	PETREK J, ZITKA O, ADAM V, et al. Are early somatic embryos of the Norway spruce (Picea abies (L.) Karst.) organised?[J]. PLoS One, 2015, 10(12): e0144093.
[15]	崔国栋. 林木悬浮细胞系建立过程中的影响因素[J]. 山西林业科技, 2010, 39(4): 35-37.
[16]	谭兴杰, 李月标. 荔枝多酚氧化酶的部分纯化及性质[J]. 植物生理学报, 1984, 10(4): 339-345.
[17]	王政, 周方方, 尚文倩, 等. 培养基成分对牡丹愈伤组织褐化的影响[M]//张启翔.中国观赏园艺研究进展. 北京: 中国林业出版社, 2015: 248-252.
[18]	邱璐, 陈善娜, 杨跃仙, 等. 云桑组织培养中褐化问题的研究[J]. 蚕业科学, 2000, 26(2): 118-119.
[19]	DONG Y S, FU C H, SU P, et al. Mechanisms and effective control of physiological browning phenomena in plant cell cultures[J]. Physiol Plant, 2016, 156(1): 13-28.
[20]	刘艺平, 王政, 牛佳佳, 等. 不同糖源及蔗糖质量浓度对牡丹愈伤组织褐化的影响[J]. 河南农业科学, 2013, 42(3): 103-106.

Cited By

Get Citation

PDF

XML

Article views PDF downloads

Turn off MathJax

Article Contents

Abstract

References

Investigation of browning factors in embryogenic callus culture of Pinus tabulaeformis and optimization of proliferation medium

Abstract

References

Catalog

Export File

Citation

Format

Content