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A comparison between mitochondrial DNA of an isogenic male-sterile (5) and male-fertile (F) couple (HA89) of sunflower

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Summary

Mitochondrial DNA from etiolated seedling of male-fertile (F) and male-sterile (S) sunflower (Helianthus annuus L.) has been investigated for the first time in an isogenic couple HA89.

  1. 1.

    The mt-DNAs isolated from the S and F cytoplasms give a sharp and symmetrical band at 1.706 g/ml in an analytical CsCI gradient ultracentrifugation.

  2. 2.

    In polyacrylamide gel it is found that mitochondria from F cytoplasm contain a Low-Molecular-Weight (LMW) mt-DNA molecule of 1.45 Kilobase-pairs (kb) in addition to the major mt-DNA. On the contrary, mitochondria from S cytoplasm contain no LMW molecule. Treatments with DNAse, RNAse and Nuclease S1 show that this LMW mt-DNA molecule consists of a supercoiled circular DNA. The length and form of the LMW molecule are confirmed by electron microscopy observations.

  3. 3.

    The mt-DNAs from S and F cytoplasms have also been investigated using restriction endonuclease digestions (Sa11, Xho1, Bg11). The electrophoresis of the resulting fragments reveals several differences between the S and F cytoplasm.

It is not known whether the differences observed between F and S mt-DNA are correlated with the cytoplasmic male sterility phenotype.

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Abbreviations

MOPS:

3-[N-morpholinojpropanesulfonic acid

EGTA:

ethyleneglycol-bis(β-amino-ethyl-ether)N,N′-tetra acetic acid

BSA:

bovine serum albumine

PVPP:

polyvinylpolypyrrolidone

EDTA:

ethylenediaminetetraacetic acid

Tris:

tris(hydroxymethyl)aminomethane

TEMED:

N,N,N′,N′-tetramethylenediamine

Buffers:

SSC: 0.15 M NaCl, 0.02M Na-citrate, pH 7.0

TNE:

0.01 M Tris, 0.05 M NaCl, 0.001 M EDTA, pH 8.0

TBE 10 x:

0.9 M Tris, 0.9 M Boric acid, 0.025 M EDTA, pH 8.3

S:

5% (w/v) Sucrose, 0.1 M EDTA, 0.02% (w/v) bromophenol blue, pH 8.0

EBV:

40 mM Tris, 2 mM EDTA, 20 mM Na-acetate, pH 7.8 (acetic acid)

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Correspondence to Philippe Leroy.

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Leroy, P., Bazetoux, S., Quétier, F. et al. A comparison between mitochondrial DNA of an isogenic male-sterile (5) and male-fertile (F) couple (HA89) of sunflower. Curr Genet 9, 245–251 (1985). https://doi.org/10.1007/BF00420319

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Key words

  • Cytoplasmic male sterility
  • Hehanthus annuus
  • mt-DNA
  • Sunflower