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Nitro-blue tetrazolium: A specific stain for photosynthetic activity in protoplasts

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Abstract

A qualitative assay for the detection of photosynthetic activity in protoplasts is described. Leaf protoplasts from atrazine resistant and susceptible biotypes of Brassica napus suspended in medium with 0.01% nitro-blue tetrazolium and 10 to 100 µM atrazine showed clear differences in staining. Staining was detectable with 0.001% fluorescein-labelled tetrazolium which, unlike nitro-blue tetrazolium, did not cause collapse of the protoplasts.

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Abbreviations

atrazine:

2-chloro-4-(ethylamino)-6(isopropyl-amino)-s-triazine

DCPIP:

dichlorophenolindophenol

fluorescein-labelled tetrazolium:

2,5-bis-(4-nitrophenyl)-3-(5-fluoresceinyl)-2H-tetrazolium chloride

Hepes:

N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid

Mes:

2-(N-morpholino)ethanesulfonic acid

nitro-blue tetrazolium:

2,2′,5,5′-tetraphenyl-(3,3′-dimethoxy-4,4′-biphenylene)-ditetrazolium chloride

PS:

0.6M sorbitol + 10mM NaHCO3 + 50mM Hepes, pH 7.6

SM:

0.5M sorbitol + 10mM Mes, pH 5.8

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Author notes

  1. Dominique Robertson

    Present address: Department of Zoology, Duke University, 27706, Durham, NC, USA

Authors and Affiliations

  1. Department of Plant Breeding, Cornell University, 14853, Ithaca, NY, USA

    Dominique Robertson & Elizabeth D. Earle

Authors
  1. Dominique Robertson
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  2. Elizabeth D. Earle
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Additional information

Communicated by O. Gamborg

This work has been submitted by D. R. in partial fulfillment of the requirement for the Ph.D. degree

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Robertson, D., Earle, E.D. Nitro-blue tetrazolium: A specific stain for photosynthetic activity in protoplasts. Plant Cell Reports 6, 70–73 (1987). https://doi.org/10.1007/BF00269743

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  • DOI: https://doi.org/10.1007/BF00269743

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