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Effects of Exogenous 2,4-Dichlorophenoxyacetic Acid, Glutathione, and L-Buthionine Sulfoximine on Intracellular Redox State and Hormone Levels and Their Links with Proliferation in Picea pungens Somatic Embryogenesis

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Abstract

Cellular redox state and hormone levels have important regulatory roles in the process of somatic embryogenesis (SE), but experimental data on the topic are limited in conifer SE. This study investigated changes in redox pairs (reduced glutathione (GSH) vs oxidized glutathione (GSSG) and ascorbic acid (ASA) vs dehydroascorbate (DHA)), activities of enzymes (glutathione reductase (GR), DHA reductase (DHAR), and ascorbate peroxidase (APX)), and levels of endogenous hormones (indole-3-acetic acid (IAA), zeatin-riboside (ZR), gibberellin (GA), and abscisic acid (ABA)) following exogenous supplements of 2,4-dichlorophenoxyacetic acid (2,4-D), GSH, and L-buthionine sulfoximine (BSO) to the proliferation medium of Picea pungens SE and linked the changes with proliferation efficiencies. While 2,4-D promoted proliferation and maturation in a concentration-dependent manner, the joint application of 2,4-D (2 mg/L) and GSH (0.5 mmol/L) was the most effective. Such beneficial roles were not observed when GSH alone was applied and exogenous BSO (1 mmol/L) prohibited proliferation. Supplements of 2,4-D and GSH, individually or jointly, enhanced the GR activity, decreased the DHAR activity and increased amounts of redox compounds, yet more in GSH and ASA than in GSSG and DHA, creating reduced environments. Exogenous BSO had reversal effects, creating an oxidized environment, and so was the joint use of 2,4-D and BSO, although at a less extent. 2,4-D and GSH reduced while BSO increased IAA content. The alternations in redox state as well as in IAA content from exogenous 2,4-D or BSO supplements coincided with the corresponding changes in proliferation. The notable finding was that exogenous GSH, regardless its substantial roles in reducing environments or IAA content, did not improve proliferation. Thus, the role of GSH on proliferation varied, depending on 2,4-D existing, and could not be fully accounted for by dynamic changes in redox state or hormone level. Alternations in APX and hormones ABA, ZR, GA, and IAA/ABA ratio were also found not highly involved in altering proliferation.

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Abbreviations

ABA:

Abscisic acid

AsA-GSH cycle:

Ascorbate–glutathione cycle

APX:

Ascorbate peroxidase

AsA:

Ascorbic acid

BSO:

L-buthionine sulfoximine

2,4-D:

2,4-Dichlorophenoxyacetic acid

DHA:

Dehydroascorbate

DHAR:

Dehydroascorbate reductase

EC:

Embryogenic calli

GA:

Gibberellin

GR:

Glutathione reductase

GSH:

Reduced glutathione

GSSG:

Oxidized glutathione

IAA:

Indole-3-acetic acid

IAAO:

Acetic acid oxidase

MDHA:

Monodehydroascorbic acid

MDHAR:

Monodehydroascorbic acid reductase

PEM:

Proembryogenic mass

PGR:

Plant growth regulator

SE:

Somatic embryogenesis

ZR:

Zeatin-riboside

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Funding

This research is supported by the Major Science and Technology Project “Research on Collection and Preservation Technologies of Germplasm Resources in the Trees of Changbai Mountain” funded by Jilin Provincial Forestry Bureau (2015–002) and the Forestry Science and Technology Promotion and Demonstration Project, “Promotion of Efficient Propagation Technologies in Blue Spruce” funded by the Central Finance of China (No. JLT2021-10).

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Authors and Affiliations

  1. Jilin Provincial Academy of Forestry Sciences, 3528 Linhe St., Changchun, 130033, Jilin, China

    Fang Gao, Fangfang Yan, Caiyun Qin, Jufeng Cai, Jing Tao & Shigang Chen

  2. College of Horticulture, Jilin Agricultural University, 2888 Xincheng St., Changchun, 130118, Jilin, China

    Fangfang Yan

  3. Forest Seedling Management Station, Jilin Provincial Forest and Grassland Department, 3698 Yatai Avenue, Changchun, 130022, Jilin, China

    Conghui Wang

  4. Arthur Temple College of Forestry and Agriculture, Stephen F. Austin State University, Nacogdoches, TX, 75962, USA

    Yuhui Weng

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  1. Fang Gao
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Contributions

Conception, F.G., F.F.Y. and S.Q.C; Writing-draft preparation, F.G., J.T., F.F.Y. and Y.H.W.; Writing-review and editing, F.G., S.G.C., J.T. and Y.H.W.; Experimental planning, F.G, J.T. and S.G.C.; Experiment set up and data collection, Fa.G., F.F.Y., C.Y.Q., J.F.C., C.B.S., C.H.W.; Data analysis, F.G., Y.H.W. All authors have read and agreed to the published version of the manuscript.

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Correspondence to Yuhui Weng, Jing Tao or Shigang Chen.

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Gao, F., Yan, F., Qin, C. et al. Effects of Exogenous 2,4-Dichlorophenoxyacetic Acid, Glutathione, and L-Buthionine Sulfoximine on Intracellular Redox State and Hormone Levels and Their Links with Proliferation in Picea pungens Somatic Embryogenesis. J Plant Growth Regul (2023). https://doi.org/10.1007/s00344-023-11198-3

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