|Budget Amount *help
¥15,200,000 (Direct Cost : ¥15,200,000)
Fiscal Year 2004 : ¥1,500,000 (Direct Cost : ¥1,500,000)
Fiscal Year 2003 : ¥1,400,000 (Direct Cost : ¥1,400,000)
Fiscal Year 2002 : ¥12,300,000 (Direct Cost : ¥12,300,000)
(1)The S-locus genes (Si-and Sz-RNase) in the pistil were doped and sequenced from the apple (Malus x domestica Borkh.) ailtivar McIntosh. We developed S-allele specific polymerase chain reaction (PCR) -digestion method for identification the S-alleles. Using the method, we analyzed S-genotype of 310 apple species, cultivars and selections. As the parents of the some of the apple cultivars were unknown and others were uncertain, we detennined accurate parents of eight apple cult ivars using SSR markers.
(2)For unification of the alphabetical S-RNase allele designation in Japan to the European numerical designations, we clarified that the S_<a^->, S_<b^->, S_<c^->, S_<d^->, S_<e^->, S_<f^->, S_<g^->, S_<h^->, S_<i^->, S_<z^->-RNase alleles, respectively, to the S_<2^->, S_<3^->, S_<9^->, S_<7^->, S_<28^->, S_<1^->, S_<20^->, S_<24^->, S_<10^-> and S_<25^->-RNase allele. We resolved the confusion of the numerical S-RNase allele designations, and established the standardized S-RNase allele
designation of apple.
(3)We isolated three cDNA clones (MdiPI, MdTM6 and MdiMADS13) homologous to class-B MADS-box protein from Malus x domesirca Borkh. 'Indo'. MdTM6 and MdiMADS13 genes showed the same expression patterns, and are close to TM6 lineage members. This is the first case of two TM6 lineage genes existing in one plant. Producing transgenic apple possessing apetalous flowers and parthenocarpic fruit is under propressing.
(4)We isolated rose MADS-box genes 'MASAKO BP, B3 and euB3' homologous to class-B floral identity genes. Functional analyses using transgenic Arabidopsis with ectopically expressed MASAKO BP and B3 or MASAKO BP and euB3 under the control of CaMV35S promoter produced flowers with sepal-to-petal and pistil-to-stamen or sepals-to-petals, suggesting that MASAKO BP and B3, and MASAKO B3 and euB3 are involved in petal and stamen identification in rose.
(5)The transgenic plants with ectopically expressed rose MADS -box genes 'MASAKO C1 and D1' homologous to class-C floral identity genes under the control of CaMV35S promoter produced flowers with sepa1-to-carpel and petal-to-stamen transformation in Arabidopsis and sepal-to-cazpel transformation in Torenia. These results suggest that both genes have similar function and are involved in stamen and carpel identity in the rose.
(6)We isolated Rose MADS-box genes 'MASAKO S1 and S3' homologous to class-E floral organ identity genes. We found that MASAKO S1 and S3 are homologues to SEPALLATA (SEP)1/2 and 3, respectively. The expression patterns and cDNA structures of MASAKO S1 and S3 in the green rose flower were the same as those in the normal flower of Rosa rugasa, suggesting that homeotic changes in the green rose flower might not develop into mutations of the MADS-box genes alone. Less