This study was conducted to investigate the effects of pretreatment and shipping temperature on leaf chlorosis in cut Lilium Oriental hybrid 'Siberia'. Cut lilies were shipped under various temperatures (5, 10, 15, $25^{\circ}C$) for 5 days. When cut lilies were shipped at $25^{\circ}C$, leaf chlorosis was accelerated. However, chlorosis was significantly decreased by shipping at 5 to $15^{\circ}C$. In addition, leaf chlorosis was significantly decreased when the cut lilies were pretreated with a solution containing Promalin (BA + $GA_{4+7}$) as compared to the control. Promalin completely prevented postharvest leaf chlorosis, whereas $GA_3$ and Chrysal SVB were ineffective. Leaf chlorosis decreased more with Promalin dip treatment than with spray treatment. This pretreatment solution also extended the vase life of cut lilies. When cut lilies were pretreated with Promalin, yield (Fv/Fm) of chlorophyll fluorescence was highly maintained. Especially chlorophyll content was significant increased by Promalin treatment. Thus, shipping between 5 and $15^{\circ}C$ and Promalin dip pretreatment significantly decreased leaf chlorosis in cut 'Siberia' lilies.
These studies were carried out to select somatic hybrid using selectable marker genes of Nicotiana glauca transformed by NPTII gene and Solanum tuberosum transformed by T- DNA, and to study characteristics of transformant. The results are summarized as follows. 1. Crown gall tumors and hairy roots were formed on potato tuber disc infected by A. tumefaciens Ach5 and A. rhizogenes ATCC15834. These tumors and roots could be grown on the phytohormone free media. 2. Callus formation from hairy root was prompted on the medium containing 2, 4 D 2mg/I with casein hydrolysate lg/l. 3. The survival ratio of crown gall tumor callus derived from potato increased on the medium containing the activated charcoal 0. 5-2. 0mg/I because of the preventions on the other hand, hairy roots were necrosis on the same medium. 4. Callus derived from hairy root were excellently grown for a short time by suspension culture on liquid medium containing 2, 4-D 2mg/I and casein hydrolysate lg/l. 5. The binary vector pGA643 was mobilized from E. coli MC1000 into wild type Agrobacteriurn tumefaciens Ach5, A. tumefaciens $A_4T$ and disarmed A. tuniefaciens LBA4404 using a triparental mating method with E. ccli HB1O1/pRK2013. Transconjugants were obtained on the minimal media containing tetracycline and kanamycin. pGA643 vectors were confirmed by electrophoresis on 0.7% agarose gel. 6. Kanamycin resistant calli were selected on the media supplemented with 2, 4-D 0.5mg/1 and kanamycin $100\mug$/ml after co- cultivating with tobacco stem explants and A. tumefaciens LBA4404/pGA643, and selected calli propagated on the same medium. 7. The multiple shoots were regenerated from kanamycin resistant calli on the MS medium containing BA 2mg/l. 8. Leaf segments of transformed shoot were able to grow vigorusly on the medium supplemented with high concentration of kanamycin $1000\mug$/ml. 9. Kanamycin resistant shoots were rooting and elongated on medium containing kanamycin $100\mug$/ml, but normal shoot were not. 10. For the production of protoplast from potato calli transformed by T-DNA and mesophyll tissue transformed by NPTII gene, the former was isolated in the enzyme mixture of 2.0% celluase Onozuka R-10, 1.0% dricelase, 1.0% macerozyme. and 0.5M mannitol, the latter was isolated in the enzyme mixture 1.0% Celluase Onozuka R-10, 0.3% macerozyme, and 0.7M mannitol. 11. The optimal concentrationn of mannitol in the enzyme mixture for high protoplast yield was 0.8M at both transformed tobacco mesophyll and potato callus. The viabilities of protoplast were shown above 90%, respectively. 12. Both tobacco mesophyll and potato callus protoplasts were fused by using PEG solution. Cell walls were regenerated on hormone free media supplemented with kanamycin after 5 days, and colonies were observed after 4 weeks culture.
The objective of this research was to produce sod by box seeding for zoysiagrass or by vegetative propagation for zoysiagrass and manilagrass.1 Various ratio of peatmoss to sand(v /v) were prepared to find idea[ medium for fast and light weight sod production. Then, the days required for sod formation, the effect of growth regulators on the growth of turfgrass, and the various storage methods for winter keeping of sods were also investigated. 1.The mixed medium of sand and peatmoss(v /v, 1 : 2) showed more biomass production than that of sand. 2.In comparison of seeding rate of zoysiagrass, the amount of log /$m^2$ was most effective in the fast and dense sod formation. The amount of 20g /$m^2$ also showed fast sod formation. But, it resulted in weak plant and less tillering. During April to June, about 100 days were required to form sod with seeding rate of 5g /$m^2$ regardless of seeding time. Whereas 80 days were required to form sod in the rate log /$m^2$, which was 20 days shorter than that of 5g /$m^2$. 3.More than 85% of shoots in sod stored in field or plastic house during the winter time resumed the growth in good appearance after transplanting. The whole covering of ground with sod resulted in less weeds and faster formation of lawn. 4.Vegetative propagation of manilagrass showed about 7 to 15 days faster formation of sod than that of zoysiagrass. Application of GA increased shoot growth and BA increased the total number of tillering. However, the effects of the combined application of GA and BA were negligable.
Loaves, stems, cotyledons, and roots of Hovenia dulcis Thunb grown in test tube were cultured on media containing different concentrations of single or combined growth regulators. In MS media containing 2mg/ι BA, the shoot formation rate was 95.5% and it was the highest frequency of shoot formation. MS media showed most efficiency in the shoot formation at 0.01mg/ι TDZ for the callus formation, but the color of callus changed to brown at a higher concentration of TDZ. Callus formation was 89.% at 0.5mg/ 2.4-D, but IAA, IBA, and NAA were not effective on the formation of callus. Calli were formed only on wound area when IAA, IBA, and NAA were added into MS media. Combined growth regulators (BA + auxin) were more effective in roots and nodes than leaves and cotyledons on the formation of shoot. More than 97% of shoot formation was obtained on MS media containing BA and auxin. For the production of multiple shoot, nodes of Hovenia dulcis were used and effect of growth regulators on the formation of multiple shoot was evaluated on MS media. Highest shoots (5.3) of Hovenia dulcis were induced on MS media supplied with 0.1mg/ι BA and 0.1mg/ι NAA, and an average of 6.4 shoots per explant were obtained in 1/2 MS media containing same concentration and growth regulators. An average of 7 shoots per explant after 4 weeks of culture from nodes of Hovenia dulcis was produced on a woody plant medium(WPM) containing 0.1mg/ι BA and 0.1mg/ι NAA. Shoot length was 6.0 cm in average.
Chrysanthemum is a cut flower species that normally lasts for 1 to 2 weeks, in some cases 3-4 weeks. This has been attributed to low ethylene production during senescence. Reduction in cut flower quality has been attributed to the formation of air embolisms that partially or completely blocks the water transport from the vase solution to the rest of the cut flower stem, increasing hydraulic resistance which may cause severe water stress, yellowing, wilting of leaf, and chlorophyll degradation. Standard type chrysanthemum can be harvested when buds were still tightly closed and then fully opened with the simple bud-opening solution. Standard type chrysanthemum can also be harvested when the minimum size of the inflorescence is about 5-6 cm bud which opened into the first flower full-sized flower. While spray varieties can be harvested when 2-4 most mature flowers have opened (40% opening). Cut flowers are sorted by stem length, weight, condition, and so on. Standard chrysanthemum is 80 cm length for standard type and 70cm for spray type. Pre-treatment with a STS, plant regulator such as GA, BA, 1-MCP, chrysal, germicide, and sucrose, significantly improved the vase life and quality of cut flowers. It is well established that vase solutions containing sugar can improve the vase life of cut chrysanthemum. Chrysanthemum is normally packed in standard horizontal fiberboard boxes. Chrysanthemum should normally be stored at $5{\sim}7^{\circ}C$. Precooling resulted in reduction in respiration, decomposition, and transpiration activities as well as decoloration retardation. There was significant difference between "wet" storage in 3 weeks and "dry" storage in 2 weeks. In separate pulsing solution trials, various germicides were tested, as well as PGRs to maintain the green color of leaves and turgidity. Prolonging vase life was attained with the application of optimal solution such as HQS, $AgNO_3$, GA, BA and sucrose. This also retarded senescence in leaves of cut flower stems. Fresh cut chrysanthemum can be transported using a refrigerated van with $5{\sim}7^{\circ}C$. Increasing consumption and usage of cut chrysanthemum of various cultivars would require efficient transport system, and effective information exchange among producer, wholesaler, and consumer.
A simple, high-frequency and reproducible protocol for induction of adventitious shoot buds and plant regeneration from leaf-disc cultures of Jatropha curcas L. has been developed. Adventitious shoot buds were induced from very young leaf explants of in vitro germinated seedlings as well as mature field-grown plants cultured on Murashige and Skoog's (MS) medium supplemented with thidiazuron (TDZ) ($2.27{\mu}M$), 6-benzylaminopurine (BA) ($2.22{\mu}M$) and indole-3-butyric acid (IBA) ($0.49{\mu}M$). The presence of TDZ in the induction medium has greater influence on the induction of adventitious shoot buds, whereas BA in the absence of TDZ promoted callus induction rather than shoot buds. Induced shoot buds were multiplied and elongated into shoots following transfer to the MS medium supplemented with BA ($4.44{\mu}M$), kinetin (Kn) ($2.33{\mu}M$), indole-3-acetic acid (IAA) ($1.43{\mu}M$), and gibberellic acid ($GA_3$) ($0.72{\mu}M$). Well-developed shoots were rooted on MS medium supplemented with IBA ($0.5{\mu}M$) after 30 days. Regenerated plants after 2 months of acclimatization were successfully transferred to the field without visible morphological variation. This protocol might find use in mass production of true-to-type plants and in production of transgenic plants through Agrobacterium/biolistic-mediated transformation.
Several seeded cultivars of Vitis labruscana showing various responses to GA-induced seedlessness were tested to study the effects of streptomycin sulfate on induction of parthenocarpy. The results were summarized as follows. 1. Prebloom dipping of streptomycin sulfate at 200 ppm induced 90~100 % parthenocarpy in 'Schuyler', 'Delaware', 'Ohira-Dela.' and 'Tanored' cultivars and also stimulated maturity but did not show any toxic effects. 2. In most cultivars, the addition of 25 ppm GA to streptomycin tended to increase berry setting and induction of parthenocarpy. 3. Dipping of strepto mycin alone in 'Campbell Early' severely induced berry abscission but the addition of calcium acetate or BA to streptomycin slightly increased setting of berries. 4. In 'Kyoho' and 'Pione' cultivars, streptomycin successfully induced parthenocarpy. Berry setting by streptomycin treatment was variable according to environmental conditions. 5. In 'Campbell Early' and 'Tanored' cultivars, streptomycin severely reduced viability of pollen.
Protoplasts were isolated from hypocotyl tissues of 5-day-old Brassica oleracea var capitata Green Challenger seedlings. Several media were used for protoplast culture and shoot regeneration. The shoot-regeneration rapacity of protoplast derived callus depended on the initial culture medium. Protoplasts were cultured in liquid medium (B5 medium supplemented with CaCl2, 2H2O 600mg/L, g1ucose 20g/L, D-mannito1 70g/L, NAA lmg/L, BA lmg/L, 2.4-D 0.25 mg/L)at 27$^{\circ}C$ under the dark After 5 to 10 days, cultlues were diluted with medium with a reduced osmotic stabilizer and then transferred to illuminated conditions. The culture medium was changed with the fresh medium at 7- to 10-day-intervals until the formation of microcallus. Hypocotyl protoplast-derived callus proliferated when transferred to MS medium supplemented with NAA lmg/L, BA 1mg/L and GA$_3$ 0.02mg/L. Upon transfer to MS basal medium without growth regulators, roots were produced. In an attempt to increase the regeneration frequency, 10g/L polyvinylpyrrolidone was added to the regeneration medium, but the shoot regeneration was mot improved. The regenerated whole plants were acclimated in a sterized soilless mixture(vermiculite 2;perlite 2;peat moss1) in a culture room.
This study describes an efficient and stable droplet vitrification following cryopreservation of strawberry shoot tip (Fragaria × ananassa Duch.) accessions 'Massey' and 'MDUS3816'. The shoot tips were precultured in Murashige and Skoog (MS) liquid medium supplemented with sucrose (0.3-0.7M). Precultured explants were osmoprotected with loading solution (LS, C4) containing 17.5% glycerol and 17.5% sucrose for 40 min and exposed to dehydration solution (B1) containing 50% glycerol and 50% sucrose for 40 min at 25oC. Subsequently, the explants were transferred onto droplets containing 2.5 µL PVS3 on sterilized aluminum foils (4 cm× 0.5 cm) prior to direct immersion in liquid nitrogen (LN) for 1 h. The highest regrowth rate (%) in both the cultivars was obtained when the shoot tips were precultured with 0.3M sucrose for 30 h + 0.5M sucrose for 16 h at 25oC. The cryopreserved shoots tips exhibited 57.8 % recovery rate by culturing in NH4NO3-free MS medium supplemented with 3% sucrose, 1.0 g/L casein, 1.0mg/L GA3, and 0.5 mg/L BA for 5 weeks and in MS medium supplemented with 0.5 mg/L GA3 for 8 weeks. Variation was not observed in both of ploidy analysis and morphological investigation on plantlets of two accessions cryopreserved under variable preculture conditions.
We describe culture conditions for a high-efficiency in vitro regeneration system of Papaver nudicaule through somatic embryogenesis and secondary somatic embryogenesis. The embryogenic callus induction rate was highest when petiole explants were cultured on Murashige and Skoog (MS) medium containing 1.0 mg $1^{-1}$${\alpha}$-naphthaleneacetic acid (NAA) and 0.1 mg $1^{-1}$ 6-benzyladenine (BA) (36.7%). When transferred to plant growth regulator (PGR)-free medium, 430 somatic embryos formed asynchronously from 90 mg of embryogenic callus in each 100-ml flask. Early-stage somatic embryos were transferred to MS medium containing 1.0 mg $1^{-1}$ BA and 1.0 mg $1^{-1}$ NAA to germinate at high frequency (97.6%). One-third-strength MS medium with 1.0% sucrose and 1.0 mg $1^{-1}$$GA_3$ had the highest frequency of plantlet conversion from somatic embryos (91.2%). Over 90% of regenerated plantlets were successfully acclimated in the greenhouse. Secondary somatic embryos were frequently induced directly when the excised hypocotyls of the primary somatic embryos were cultured on MS medium without PGRs. Sucrose concentration significantly affected the induction of secondary embryos. The highest induction rate (89.5) and number of secondary somatic embryos per explant (9.3) were obtained by 1% sucrose. Most secondary embryos (87.2-94.3%) developed into the cotyledonary stage on induction medium. All cotyledonary secondary embryos were converted into plantlets both in liquid and on semisolid 1/3-strength MS medium with 1.0% sucrose.
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