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Effects of antioxidants on the plant regeneration and GUS expressive frequency of peanut (Arachis hypogaea) explants by Agrobacterium tumefaciens

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Abstract

Our objective is to develop an Agrobacterium-based transformation system for peanut. Ascorbic acid (AA), sodium selenite (Se), DL-α-tocopherol (TOC) and glutathione (GSH) were used as antioxidants during the plant regeneration and co-cultivation with Agrobacterium tumefaciens. Percentage of explants with buds or shoots increased from 50 in control group to 88, 90, 87 and 76 in GSH, TOC, Se or AA treated groups, respectively. The percentage of GUS-positive plantlets increased from 3.9 (in control) to 14.5, 10.3, 12.4 and 3.9 in GSH, TOC, Se or AA groups, respectively. Some of the callus in AA group became brown and died 2 months later. GSH, TOC and Se not only eliminated the formation of H2O2 produced in wound tissue during preparation of leaflets and co-cultivation with Agrobacterium tumefaciens and decreased malondialdehyde (MDA) formation, but also enhanced superoxide dismutase (SOD) and catalase (CAT) activities. As a result, GSH, TOC or Se increased the frequency of plant regeneration and transformation efficiency of peanut (Arachis hypogaea L.) explants by Agrobacterium tumefaciens. AA is an unsuitable antioxidant in Murashige and Skoog (MS) medium due to the stimulation of oxidation in the presence of iron in MS medium.

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Abbreviations

AA:

ascorbic acid

BA:

6-benzylaminopurine

EDTA:

ethylenediaminetetraacetic acid

GSH:

glutathione

GUS:

β-glucuronidase

HR:

hypersensitive response

MS medium:

Murashige and Skoog medium (1962)

MDA:

malondialdehyde

NAA:

1-naphthaleneacetic acid

NPTII:

neomycin phosphotransferase II

Se:

sodium selenite

SR:

shoot regeneration

TDZ:

thidiazuron

TOC:

DL-α-tocopherol

TBA:

thiobarbituric acid

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Authors and Affiliations

  1. Ocean School, Yantai University, Yantai, People’s Republic of China

    Zheng Qiusheng, Ju Bao, Liang Likun & Xiao Xianhua

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  1. Zheng Qiusheng
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  2. Ju Bao
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  3. Liang Likun
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  4. Xiao Xianhua
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Correspondence to Zheng Qiusheng.

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Qiusheng, Z., Bao, J., Likun, L. et al. Effects of antioxidants on the plant regeneration and GUS expressive frequency of peanut (Arachis hypogaea) explants by Agrobacterium tumefaciens. Plant Cell Tiss Organ Cult 81, 83–90 (2005). https://doi.org/10.1007/s11240-004-3176-9

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