Home | Vol. 19 >B. Research Notes>IV. Genetics of physiological traits and others |
25. | High-resolution mapping of glup3 gene accumulating high amount of glutelin precursor |
T. KUMAMARU1, Y. UEMURA1, Y. TAKEMOTO1, M. OGAWA2
and H. SATOH1 1) Faculty of Agriculture, Kyushu University, Hakozaki, Fukuoka, 812-8581 Japan 2) Faculty of Human Life Science, Yamaguchi Prefectural University, Sakurabatake, Yamaguchi, 753-8502 Japan |
The major storage proteins of rice consist of 40 kDa acidic and 20 kDa
basic glutelin subunits, 26 kDa globulin polypeptide, and 16 kDa, 14 kDa,
13 kDa and 10 kDa prolamin polypeptides (Tanaka et al. 1980). Seed
protein extracted from glup3 mutant lines, which were introduced
from North Asian countries and preserved in Kyushu University, contain
higher quantities of 57 kDa polypeptides and lower levels of 40 kDa acidic
and 20 kDa basic glutelin subunits compared to Taichung 65 (Satoh et
al. 1995). Kasalath. The locus of glup3 was mapped within 5.4 cM region,
between RFLP markers R374 (81.9 cM position from short arm end) and R1783
(87.3cM). For high-resolution mapping, about 2,200 glup3 homozygous
plants from F2 were selected. The genome DNA was extracted from the seedling
of glup3 homozygous plants. Using CAPS markers C377 (74.5cM) and
R3261 (87.3cM), 45 recombinants in the side of short arm and 88 in the
side of long arm were selected. From the above recombinants, 2 between
glup3 and RFLP marker L1091 and 8 between glup3 and R2783
were selected. The results indicate that Glup3 locus is located
within 0.5cM region, between L1091 and R2783. The search of BAC clone
database revealed that L1091 and R2783 corresponded to BAC clones OSJNBa0091D06
and OSJNBa0011L07, respectively. Both clones overlapped. This shows that
Glup3 gene sequence is contained within these BAC clones. To shorten
the region containing Glup3 gene, CAPS markers from the sequence
of OSJNBa0091D06 were constructed and used for further mapping of Glup3
gene. As a result, we obtained several recombinant plants (Fig. 1). These
results showed that Glup3 gene was contained in 95kb, from L1091
to long arm side. The high homologous sequence with vacuolar processing
enzyme in castor bean, soybean and Arabidopsis, which is related
to maturation of the protein in the vacuole (Hara-Nishimura et al.
1993, Kinoshita et al. 1995, Shimada et al. 1994), was detected.
The sequence has been considered as the candidate of Glup3 gene.
The complementation test by transformation with Glup3 candidate
gene is currently underway. |
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