Viçosa, Brazil
Universidade Federal de Viçosa
B chromosome derivatives in maize somatic cells
--Carvalho, CR, Saraiva, LS and Almeida, PM
Accessory or B chromosomes have been described in hundreds of plant and
animal species. They are typically smaller than the members of the regular
complement, mostly heterochromatic, transmitted in a non-Mendelian pattern,
and not necessary for normal development and reproduction. The maize B
chromosome is responsible for its own non-disjunction, due to the induction
of late knob replication during mitotic anaphase, when the heterochromatin
adjacent to the centromere remains functionally non-divided. The morphology
of the B chromosome in the root tip was analyzed. The stock used was the
Black Mexican Sweet Corn inbred line with B chromosomes from the Maize
Stock Center, at the University of Illinois. Seeds were germinated in Petri
dishes containing a film of distilled water, and incubated at 29 C in the
dark. Seedlings (1.5 to 2 cm root length) were transferred to a plastic
mesh adapted inside 6 cm diameter plastic vessels containing 3 ml of 5
µM amiprophos-methyl (APM) solution. After 2h 30 min, the roots were
washed for 15 min in running tap water, fixed in a fresh ice-cold methanol:acetic
acid solution (3:1), and kept in a freezer for at least 24 h. Subsequently,
root tip meristems were excised, macerated with 200 µL of freshly
prepared Flaxzyme (NOVO) enzymatic solution plus 1.6 ml distilled water,
and incubated at 34 C for 2h 30 min. The macerated cells were dissociated
on a clean slide with a fresh fixative solution, air-dried and stained
with a 3% Giemsa solution. Metaphasic figures were digitized directly from
a microscope-coupled CCD video camera to a computer. The karyogram was
elaborated (Figure 1) and the average length of the homologue A pairs and
B set was determined in micrometers (Table 1). Interestingly, the data
show that B chromosomes were longer than those measurements reported by
other authors. On top of that, figures of the somatic cell-derived B chromosomes
have not been widely documented. This makes any kind of comparison with
the present results difficult. Besides B chromosomes with standard morphology
(Bs), B derivatives (Bd) were observed that were
reduced in length (Figure 1). It is possible that the different morphologies
observed were due to the highly heterochromatic nature of the maize B chromosome
as affected by the arresting agent (APM) used.
Figure
1. Maize karyogram of the 2n =20A+4B chromosomes. Note the different
B chromosome morphologies. Bar = 5 µm.
Table 1. Measurements of maize root tips metaphasic A and B chromosomes.
A and B Maize chromosomes |
A Pairs |
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
9 |
10 |
Mean Length (µm) |
2.22 |
1.99 |
1.98 |
1.86 |
1.39 |
1.49 |
1.41 |
1.4 |
1.32 |
1.11 |
B Types |
Bs |
Bd1 |
Bd2 |
Bd3 |
|
|
|
|
|
|
Length (µm) |
1.48 |
1.20 |
0.91 |
0.89 |
|
|
|
|
|
|
Bs = Standard B chromosomes
Bd = Derivative B chromosomes
Please Note: Notes submitted to the Maize Genetics Cooperation Newsletter may be cited only with consent of the authors.
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