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.
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.
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.
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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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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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DOI: https://doi.org/10.1007/BF00420319