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

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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

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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

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

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