Several kinds of molecular markers such as RFLP, RAPD, STS, AFLP and
microsatellite markers, have been developed and applied in plant genetics
and breeding. Of these, microsatellite markers are valuable because they
are co-dominant, can detect high levels of allelic diversity and are economically
assayed by PCR. In rice, molecular linkage map consisting of more than
500 microsatellite markers are now available to the public (Temnykh et
al. 2001). These markers are expected to be efficiently used for QTL
analysis and markerassisted selection. In this study, molecular linkage
maps between Oryza sativa and O. rufipogon were constructed
using microsatellite markers, to confirm the utility of microsatellite
markers in studies using wild species of rice.
A single wild Myanmar accession (O. rufipogon W630) was crossed
with two O. sativa
rice cultivars Japonica Nipponbare and Indica IR36). Two BC1 mapping
populations (W630/Nipponbare//Nipponbare and W630/IR36//IR36) consisting
of 79 and 91 plants, respectively, were produced. Total DNA was extracted
from these plants. PCR amplification and band detection were carried out
using the method of Panaud et al. (1996). In total, 112 and 119
microsatellite markers were examined for Nipponbare and IR36 background
populations. The markers were selected from 12 rice linkage groups (Temnykh
et al. 2000) with an average marker distance of ca. 20 cM. Of these,
106 (94.6%) and 106 (89.1%) markers showed obvious polymorphisms between
mapping parents; these were used to check the segregation in BC1 populations.
For most of the markers, segregation followed the expected ratio of 1:1
(P < 0.05). However, 14 (13.2%) and 10 (9.4%) markers showed
significantly deviated segregation in Nipponbare and IR36 background populations,
respectively. Linkage analysis was carried out with these polymorphic
markers using MapMaker ver. 2.0 (Lander et al. 1987).
Two interspecific linkage maps were constructed between Nipponbare and
O. rufipogon W630, and between IR36 and O. rufipogon W630
(Fig. 1). Total map size were 1505 cM and 1662 cM, respectively. The marker
order of the microsatellites was consistent with that reported by Temnykh
et al. (2000), using double-haploid population between IR64 (O.
sativa Indica) and Azucena (O. sativa Javanica). To examine
the size of 12 linkage groups between interspecific and intraspecific
maps, the maximum common interval size between two markers shared by both
maps was used. Table 1 summarizes the respective length of 12 chromosomes
compared between two maps. Majority of the linkage groups showed similar
interval size. However, some showed either a relatively short or long
map size. This was probably due to a small number of markers used to compare
maps.
The present results suggest that most microsatellite markers can be used
with O. rufipogon and applied for map-based studies, such as gene
tagging and interspecific QTL analysis.
References
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