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Triticeae Genomics and Genetics
TGG 2010, Vol.1, No.1
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amplify a 2 300 bp band in ms (Ae. bicornis)-90-110
while in Ae. bicornis the specific band was absent
(Figure 2E). In the CMS lines (nuclear-cytoplasm
substitution lines), nuclear genome of the Aegilops
was substituted in the successive backcross with
common wheat 90-110. Above results indicate that
mtDNA variation in the donors of cytoplasms and
CMS lines result from genetic interactions between
90-110 nuclear genes and Aegilops cytoplasm, and
have affected the structure of the mitochondrial
genome. The structural alternations of the
mitochondrial genome are possible factors to cause
the mitochondrial defect and CMS.
Figure 1 RAPD patterns amplified by primer OPY-01, S32 and
S22 using mtDNA as template, respectively
Note: A: RAPD patterns amplified by primer OPY-01; 1: Ae.
kotschyi; 2: ms (Ae. kotschyi)-90-110, 3: ms (Ae.
kotschyi)-90-110×5253; B: RAPD patterns amplified by primer
S32; 1: Ae. variabilis; 2: ms (Ae. variabilis)-90-110; 3: ms (Ae.
variabilis)-90-110×5253; C: RAPD patterns amplified by
primer S22; 1: Ae. Ventricosa; 2: ms (Ae. ventricosa)-90-110; 3:
ms (Ae. ventricosa)-90-110×5253; M: λ-EcoT14
Ⅰ
marker;
Polymorphism patterns existed between Aegilops and male
sterile lines are marked by arrows
2.2 RAPD analysis of CMS lines and
fertility-restored hybrids
To assess the effect of fertility restoration on
mitochondrial genome, CMS plants were crossed with
the restorer line of common wheat 5253 to obtain
fertile F1 hybrids. Subsequently, remarkable
differences were found in the amplification patterns of
mtDNA between CMS lines and fertile F1 hybrids. Of
the 120 RAPD primers used, four primers OPY-01,
OPD-05, S21 and OPP-02 showed stable
polymorphisms in the male sterile lines and their
respective fertility-restored F1 hybrids. In male sterile
line ms (Ae. kotschyi)-90-110, primer OPY-01 can
amplify a specific 580 bp band while in ms (Ae.
kotschyi)-90-110×5253 (fertility-restored hybrids) the
primer can amplify a specific 550 bp band (Figure 1A).
In ms (Ae. variabilis)-90-110, primer OPD-05 can
amplify a specific 2 200 bp band, while in ms (Ae.
variabilis)-90-110×5253 the band was absent (Figure
3A). In ms (Ae. ventricosa)-90-110×5253, primer S21
can amplify two specific bands of 2 020 bp and 1 610
bp, respectively, while in male sterile line ms (Ae.
ventricosa)-90-110 the spceific bands were absent
(Figure
3B).
Similarly,
in
ms
(Ae.
bicornis)-90-110×5253, primer OPP-02 can amplify a
2 530 bp band while in ms (Ae. bicornis)-90-110 the
specific band was absent (Figure 3C). These results
suggested that the mitochondrial genomes were
altered in fertility restoration by the combination of
fertility restorer gene(s), and fertility restoration
involving a strong influence of nuclear restorer genes
on mtDNA organization. Variation of mitochondrial
genome might be dependent on different nuclear
backgrounds.
Figure 2 RAPD patterns amplified by primer S202, OPB-05, OPA-04, S153 and S21 using mtDNA as template, respectively
Note: A, B, C and D are RAPD patterns amplified by primer S202, OPB-05, OPA-04, S153 and S21 using mtDNA as template,
respectively; 1: Ae. bicornis; 2: ms (Ae. bicornis)-90-110; 3: ms (Ae. bicornis)-90-110×5253; M: λ-EcoT14
Ⅰ
marker; Polymorphism
patterns existed between Ae. bicornis and ms (Ae. bicornis)-90-110 are marked by arrows