Fluorescence in situ hybridization (FISH) is a powerful tool for the detection of DNA sequences in the specific region of the chromosomes. As well as for the integrated physical mapping, FISH karyotype analysis has to be preceded. The detailed karyotypes of two onion cultivars, which are resources for onion genome sequencing project (‘Eumginara’ and ‘Sinsunhwang’), were constructed based on triple color fluorescence in situ hybridization (FISH) using 5S rDNA, 45S rDNA, and tandem repeat sequence. All used our materials showed 2n=2x=16 with x=8 as basic chromosome number. 5S rDNAs were located on 4 loci in one pair of interstitial region of short arm chromosome in both onion cultivars. Two pairs of 45S rDNAs were positioned in distal region of short arm chromosome in ‘Eumginara’. Otherwise 5 loci of 45S rDNAs were located in distal region of two pairs of short arm chromosome in ‘Sinsunhwang’. Among them, two signals of 45S rDNAs were co-localized in distal part of short arm and long arm chromosome, respectively. In case of tandem repeat sequence was detected on telomeric region of 8 pairs of chromosomes except on 45S ribosomal DNA sites. These results will provide a valuable background for physical mapping and help to further more understand the genome sequencing project in Allium cepa.

Aquaporin5 (AQP5), a water channel plays an important role in the fluid homeostasis and cell volume control in epithelial cells. In an effort to understand fluid homeostasis in the oviduct, tissue specific expression of AQP 5 was examined together with hormonal regulation of AQP5 in the mouse oviduct. To understand the oviductal fluid homeostasis and its regulation by sex steroids, We examined AQP5 expression in mouse oviduct during developmental stage and estrous cycle, and in estrogen receptor α (ERα) knockout mice oviduct. In immature mouse oviduct, expression of AQP5 expression was examined after stimulation with gonadotropins. The effect of ERα agonist (PPT) and ERβ agonist (DPN) on the oviductal expression of AQP5 was examined in ovariectomized mouse. All samples were subjected to realtime-PCR and immunohistochemistry analysis. In oviduct epithelium, AQP5 was largely found in the apicolateral membrane and cytoplasm of ERα-positive non-ciliated cells but weakly expressed in the ciliated cells. Interstitial cells, muscle cells and blood vessels were also weakly positive for AQP5 immunoreactivity. In cyclic female mice oviductal AQP5 mRNA levels were the highest at estrous. In immature mouse oviduct AQP5 mRNA and epithelial immunoreactivity were increased by PMSG, and followed by a decrease after hCG. In ERα KO mice oviduct, AQP5 mRNA levels were significantly lower than those of WT females at diestrous stage. In immature and OVX mouse oviducts, AQP5 mRNA and epithelial immunoreactivity were significantly increased by E2 and PPT. Together, our results suggest the pivotal role of AQP5 in fluid secretion and absorption of water in non-ciliated cells in oviduct. AQP5 gene is tightly activated by estrogen – ERα signaling in non-ciliated cells in oviductal epithelium, mediating the effect of estrogen on gamete transport, fertilization and early embryo development via regulating the fluid homeostasis in oviduct.

Abscission is an important developmental process used to shed organs such as leaves, flowers and fruits. Despite the detailed characterization of growth dynamics and hormonal balance during the early steps of fruit development, the molecular aspects remain unclear. Abscission of young fruit occurs by separation of cells in anatomically distinct regions between the pedicel and junction. Differences of gene expression between central pedicel and lateral pedicel were investigated by NGS. Partial cDNAs from 15 clones from both the central pedicel and lateral pedicel were selected for nucleotide sequence determination and homology searches, and 12 clones were subsequently selected for further analysis. In preliminary series of Real Time PCR analysis, 9 genes were confirmed as showing a higher expression level in lateral pedicel than in central pedicel. Many of these genes are expressed in a central or lateral pedicel in specific manner, and the expression profiles of the representative genes were confirmed. To clarify the mechanism of MdIAA14 transcription factor gene underlying abscission zone development, we are investigating the expression patterns between central and lateral pedicels in different apple cultivar using real-time PCR and constructing the vector for transformation into apple.

The precise, fast, and cost-effective identification of important fruit crop cultivars is essential for practical breeding and plant breeder’s rights. Traditional methods for identification of persimmon cultivars are based on the evaluation of sets of morphological characteristics. However, the identification using only morphological traits is difficult to distinguish among genetically closely related cultivars. This study was conducted to develop more reliable DNA markers for identification of the 32 persimmon cultivars in Korea and Japan. In total, 309 random amplified polymorphic DNA (RAPD) markers were identified using 40 different random primers. The 4 (OPP-08) to 14 (UBD159) polymorphic bands were detected with an average of 7.7. The resulting 57 RAPD fragments were selected, and their sequences were determined for developing sequence-characterized amplified region (SCAR) markers. As a result, 15 of 57 RAPD fragments were successfully converted to SCAR markers. A single polymorphic band of the same size as the RAPD fragments or smaller DNA fragments were amplified depending on primer combinations in the 15 SCAR markers. Among these markers, a combination of eight SCAR markers (PS225_200, PSN05_420, PSF13_523, PSN11_540, PS372_567, PS485_569, PSP08_635, and PS631_735) provided sufficient polymorphisms to identify 32 persimmon cultivars depending on number and size of amplicons. These newly developed markers will be useful as a fast and reliable tool to identify persimmon cultivars.

The objective of this study was to determine the fatty acid composition of newly developed tissues of germinated soybean seeds. Five soybean accessions with varied fatty acid composition were allowed to germinate in sand under greenhouse conditions. Seedlings were picked up after 4, 6, 8, 10 and 12 days of germination and freeze dried. The fatty acid composition of the newly developed tissues was analyzed by gas chromatography. Significant variation in fatty acid composition was observed between accessions, days of germination, and variety × day of germination in whole and the cotyledons. In the case of newly developed five tissues, significant variation in fatty acid composition were observed between days of germination except oleic acid for root, hypocotyl and epicotyl stem and except stearic acid for hypocotyl and unifoliate leaves while all the parameters were significantly different for accession. Significant interactions of accession and days of germination were observed for palmitic, linoleic and linolenic acid in all tissues; only for oleic acid in hypocotyl, epicotyl and unifoliate leaves; and only for stearic acid in root, hypocotyl, epicotyl and unifoliate leaves. During germination, the fatty acid composition of newly developed tissues changed dramatically but whole seedlings and cotyledons changed slightly. These tissues contained five major fatty acids as found in original seeds, but compositions were totally different from that of the seed: higher in palmitic, stearic and linolenic acid and lower in oleic and linoleic acid. New tissues conserved their fatty acid compositions regardless of genotypic variation in the original seeds.

The ripening behavior of three apple cultivars, ‘Tsugaru’, ‘Hongro’ and ‘Fuji’ was distinctive and the involvement of POLYGALACTURONASE(PG) in the fruit softening process was confirmed to be ethylene dependent. Fruit softening is genetically coordinated by the action of several cell wall enzymes, including PG which depolymerizes cell wall pectin. Also, loss of firmness is associated with increasing of the ripening hormone, ethylene. In this work, climacteric ripening of three apple cultivars, Tsugaru, Hongro and Fuji, producing different ethylene levels and ripening responses, was examined. Correspondingly in Fuji, a linear and basal ethylene level was observed over the entire period of measurements, and Tsugaru and Hongro displayed a typical climacteric rise in ethylene production. Transcript accumulation of genes involved in ethylene biosynthesis (MdACS3 and MdACO1) and MdPG1 was studied in Tsugaru, Hongro and Fuji cultivars. Expression of MdACO1 transcripts was shown in all three ripened apple fruits. However, the MdACS3 and MdPG1 were transcribed differently in these cultivars. Comparing the MdPG1 of ‘Tsugaru’, ‘Hongro’ and ‘Fuji’, structural difference was discovered by genomic Southern analysis. Overall results pointed out that MdACS3 and MdPG1 play an important role in regulation of fruit ripening in apple cultivar.

Dracocephalum argunense Fischer ex Link (Labiatae) is a perennial herbaceous plant used as valuable materials for ornamentals, honey production, and pharmaceutics. Since seed germination of this species was quite difficult, present studies were conducted to improve the germination rate by subjecting the seeds to various environmental conditions (temperature and light) and treatments (scarification, priming and seed coating). Optimum temperature for adequate germination was 20℃ though it ranged from 15℃ to 25℃, and low temperature treatment improved germination rate. Light was required for higher germination rate in this species. The scarification of seeds resulted in much higher germination, especially by the physical treatment with sandpaper or chemical treatment with sulfuric acid for 30 seconds. Various primers with different concentrations were treated on the seeds and it was demonstrated that low temperature enhanced germination rate, regardless of kinds and concentrations of the primers. Three treatment combinations of the primers, 0.5 mM GA3 treated for 48 hours, 0.5 mM IAA for 24 hours, and 1.0 mM IAA for 24 hours, increased the seed germination rate profoundly. Soaking treatment of inorganic salts, KNO3 and KH3PO4, promoted germination when seeds were subjected to low temperature. Water soluble primers such as sucrose at 0.5 and 3% concentration and solid primer talc powder were effective in enhancing germination rate.

This experiments were carried out to find out the effects of different explant materials, kinds and concentration of plant growth regulators, and total nitrogen and sucrose contents on the in vitro regeneration of Abeliophyllum distichum Nakai. The effects of growth regulators on regeneration from 3 explant sources (leaf, internode and node) were more or less same. Leaf explants produced only callus with 2ip (Isopentenyladenine) and NAA (Naphthaleneacetic acid) treatment and other regulators had no effects. Test with internode explants yielded about same results but callus was obtained with 2,4-D (2,4-Dichlorophenoxyacetic acid). Node explants resulted in shoot regeneration by all regulator treatment except NAA and 2,4-D, but control also showed similar results. Callus formation from internode and node explants was vigorous by 2ip, zeatin, and 2,4-D treatments and high NAA concentration resulted in higher callus formation. In this experiment, various mixed treatment of growth regulators were also employed, using node as explant material. Shoot regeneration was obtained with BA (Benzyl adenine) + NAA treatments but the results were comparable with control. Generally shoot and root regeneration was poor with all combined treatment except 2ip + NAA and 2,4-D + NAA. However, callus was formed readily with all treatments. In this experiment, combined treatments of regulators were applied on the callus derived from singular regulator treatment. The results showed no shoot and root regeneration with any combination of 2,4-D, IAA (Indoleacetic acid) and NAA, but soft milky white callus was formed in all the treatments. No shoot and root regeneration was observed with any combination of 2iP, NAA and IAA, but somewhat hard, light green callus was formed in all the treatments. Callus formation decreased with high kinetin concentration in case of kinetin + NAA treatment. The experiments with total nitrogen content of media showed that low concentrations of 15 and 30mM were effective for the shoot and root regeneration. Sucrose experiment demonstrated shoot regeneration with 1~4% concentration, and root and callus formation with 2~4%. No root and callus formation was observed with 0 and 1% sucrose.

Oil and fatty acid composition of 648 soybean germplasms of different categories including Korean and American source were analyzed by NIRS (Near Infrared Reflectance Spectrophotometer) method at Yeongnam Agricultural Research Institute, Milyang, Korea. A

The effect of plant growth regulators was investigated on in vitro shoot proliferation from axillary bud explants of Hypericum erectum. To determine the optimal cytokinin for proliferation of axillay buds, we carried out screening four cytokinins (BA, kinetin, 2iP, TDZ). When nodal segments were cultured on MS medium supplemented with 4.5 μM TDZ (thidiazuron), a number of shoots were induced. Our results indicated that the addition of TDZ to culture medium resulted in the induction of significantly more axillary buds than in the addition of other cytokinins. The optimal concentration of TDZ for proliferation of axillary buds was 10 μM. 92% of shoots spontaneously rooted without any plant growth regulator (PGR) and formed whole plantlets within one month. More than 95% of these regenerants survived and they did not show any detectable variation in morphology or growth characteristics compared to their donor plants.