Calla lilies (Zantedeschia spp.) are monocotyledonous ornamental plants which belongs to the Araceae family. After the release of elite calla cultivar, an efficient propagation system is needed for commercial use. Despite the use of conventional propagation methods such as splitting of tubers and rhizomes of calla, rapid and efficient propagation system should be developed. In order to achieve this goal, stem segments contained apical meristems derived from calla lily cultivar (cv. Gag-si) were cultured on Murashige and Skoog (MS) medium supplemented with various concentrations of cytokinin and auxin. This was aimed at inducing embryogenic calluses, shoots and multiple shoots. As a result, about 25% of induction rates of yellow embryogenic calluses were observed with MS medium containing both $0.5mg{\cdot}L^{-1}\;NAA$ and $1.5mg{\cdot}L^{-1}\;BA$ as growth regulators. In the experiments involving the regeneration from embryogenic calluses through shoot formation, MS medium supplemented with $0.5mg{\cdot}L^{-1}\;IAA$ and $2.0mg{\cdot}L^{-1}\;BA$ showed the highest rates at approximately 85 ~ 90% with regard to the formation of shoots in calla. Moreover, multiple shoots needed for rapid propagation were generated when explants were cultured on MS medium supplemented with $0.5mg{\cdot}L^{-1}\;IAA$ and $2.0mg{\cdot}L^{-1}\;BA$ with 40% of formation rate. In this study, the combination of auxin and cytokinin showed positive effects on both the induction of embryogenic calluses, the formation of shoots as well as multiple shoots in calla. The regeneration system described here can contribute to the development of breeding programs of calla in the future.
Lee Ki-Jung;Seo Jen-Kyung;Lee Hye-Young;Jeon Eun-Hee;Shin Sang-Hyun;Lee Jai-Heon;Kim Doh-Hoon;Ko Jong-Min;Hahn Won Young;Baek In-Youl;Oh Boung-Jun;Chung Young-Soo
Journal of Life Science
/
v.16
no.2
s.75
/
pp.289-296
/
2006
In order to establish highly efficient gene transfer condition at early stage of soybean transformation, various experiments were performed and compared their efficiencies by transient GUS analysis; those conditions are genotype determination of Korean soybean cultivars for amenability to Agro-infection, appropriate agar and selective agent concentration, orientation of explant placement, hormone pre-culture, and liquid selection condition. In the genotype screen of Korean soybean varieties, 14 amenable genotypes were selected. For efficient Agrobacterium washing, cefotaxime was chosen and hygromycin at the concentration of 10 and 15 ppm was used as selection agent in the media. Agar concentration was slightly better in 0.6% and 0.8% for both shoot and callus formation, and explant placement with adaxial side down showed high frequency of GUS expression. For wounding treatment, oriental needle was efficient than scalpel for shoot formation and gene transfer. To increase the frequency of gene transfer, hormone pre-treatment was applied. BA at the concentration of 5 and 10 ppm resulted in better survival at the late stage of selection in shoot elongation media. Selection in liquid media after hormone pre-treatment seemed to be effective to remove the escaped non-transformants at early stage of procedure. Considering the results obtained, Eunhakong could be the first choice as a material for soybean transformation among Korean soybean genotypes.
Objective of this study was to select ectomycorrhizal fungi for black spruce(Picea mariana) inoculation to overcome the growth inhibitory effects of Kalmia angustifolia. Nineteen isolates representing 11 species of ectomycorrhizal fungi were tested for their abilities to grow and form mycorrhizae with black spruce seedlings in the presence of water leachate of leaves of Kalmia. Mycelium growth of 9 isolates were inhibited by the leaf leachate. Colony diameter and biomass of the other 10 isolates were either increased or unaffected under the same conditions. Acidic pH of the culture medium(pH 3 and 4) inhibited some of the fungi, but a combination of acidic pH and the leaf leachate was more inhibitory. Thirteen isolates were able to form ectomycorrhizae with black spruce in presence of 25% leaf leachate in pure culture. Four isolates, Paxillus involutus(NF4), Cenococcum geophilum(GB12), Laccaria laccata(GB23), and E-strain(GB45) formed mycorrhizae more successfully than the others in presence of up to 50% Kalmia leaf leachate. Black spruce seedlings pre-inoculated with these fungi were grown with Kalmia leaf leachate and live Kalmia plants during a four month greenhouse experiment. Abundant mycorrhizae(77-91% of root tips) were developed on seedlings pre-inoculated with P. involutus, L. laccata and E-strain but relatively poor mycorrhization(32% of root tips) resulted with C. geophidum. Over 90% of the short root mycorrhizae were attributed to the inoculated fungi although indigenous mycorrhizae also occurred on most seedlings. Persistence of the mycorrhizae was not affected by living Kalmia plants. Over 80% of the mycorrhizae on seedlings inoculated with P. involutus, L. laccata and E-strain and 53% of the mycorrhizae on seedlings inoculated with C. geophilum were attributable to the inoculant fungi. Control seedlings formed about 45% ectomycorrhizal short roots with indigenous fungi. The L. laccata and C. geophilum inoculated seedlings exhibited enhanced mycorrhizae formation in presence of Kalmia leaf leachate. Mycorrhizae formation with inoculant fungi was 4-15% lower at pH 4 than at pH 5, with the greatest inhibition occurring for L. laccata. Seedlings inoculated with P. involutus had the greatest shoot and root growth followed by L. laccata and E-strain inoculated seedlings. The P. involutus and L. laccata inoculated seedlings were significantly taller with more shoot dry biomass than the uninoculated(control) seedlings. E-strain inoculated seedlings had significantly higher shoot dry biomass and significantly lower number of first order lateral roots compared to the control but other growth parameters such as height, root dry weight and number of short root tips were not significantly different from the control. Seedlings inoculated with C. geophilum were not significantly different from the uninoculated seedlings in any of the growth parameters except for the number of first artier lateral roots which was significantly less than the control seedlings.
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.
An experiment was conducted to find out effect of soil water condition on shoot and root growth and P, K uptake by soybean plants. Soybean plants were grown under different soil water table levels, 20cm, 40cm and 60cm below the soil surface using minirhizotron with 20cm in diameter, and well irrigated and water stressed conditions using 1/2000a Waganer pots. Three soybean plants, Paldal culfivar, were grown and sampled at the early growing period, 37 days after planting, and at the harvesing period, 115 days after planting. Shoot and root growth were restricted by water stresed condition and by excessive soil water condition with the 20cm water table. Little difference in shoot and root growth were found between well irrigated condition and 40cm or 60cm water table conditions. The P and K contents in shoot under water stressed condition were higher than well irrigated condition at the early growing period but reversed at later harvesting period. The dry weight and length of roots were more severely restricted by water stress than those of shoots. Root morphological difference was found by anatomical observation. Normal cortex was developed under the well irrigated condition, while abnormal cortex with aerenchyma formed by lysis under excessive water conditions of 20 or 40cm water talbes. Aerenchyma was formed at outer skirt of pericycle. Role of formation of aerenchyma of soybean roots might be an adaptation to excessive soil water condition and possibly related to survival of soybean roots growing under near flooding condition on dikes of paddy lands.
The repeated subcultures of in vitro plant materials in wasabi became highly vitrified and the capacity for multiple shoot formation from the vitrified plant materials was very low. In order to improve the quality of in vitro propagated planting materials, the experiments were carried out using culture vessels capped with membrane filter(MF). When vitrified shoots were cultured on MS medium with 0.2mg/L BA in the vessels with MF or without MF for 60 days, the shoots in the vessels with MF did not vitrified. In contrast, the shoots grown in the vessels without MF vitrified at 65%. The stomates of vitrified leaves were circular and inflated, whereas those of normal leaves acclimatizated in the vessels with MF were ovate in shape. The hardened shoots were also cultured on MS media without sucrose containing 0.01mg/L IBA in vessels with(photoautotrophic culture) or without(control) MF. Sucrose was necessary for survival of the in vitro plantlets in the vessels without MF. After 20 days of culture, the shoots in the vessels without MF on the sucrose-free media turned yellow and died. But the shoots in the vessels with MF in the sucrose-free media produced a lot of roots. When shoots were cultured on MS medium with 2% sucrose containing 0.01mg/L IBA in the vessels with(photomixotrophic culture) or without(heterotrophic culture) MF, best growth occured in photomixotrophic culture.
In plants, a shoot apex has a small region known as the shoot apical meristem (SAM) having a group of dividing (initiating) cells. The SAM gives rise to all the groundabove structures of plants throughout their lifetime, and thus it plays important role in growth and development of plants. This review describes theories to explain the SAM organization and function developed over the last 250 years. Since in 1759 German botanist C. F. Wolff has described firstly the SAM, in 1858 Swiss botanist C. N${\ddot{a}}$geli proposed the apical cell theory from the observation of a large single apical cell in the SAM of seedless vascular plants: however, this view was recognized to be unsuitable to seed plants. In 1868, German botanist J. Hanstein suggested the histogen theory: this concept subdividing the SAM into dermatogen, periblem, and plerome was unable to generally apply to seed plants. In 1924, German botanist A. Schmidt proposed the tunica-corpus theory from the examination of angiosperm SAM in which two parts show different planes of cell division: this theory was proved to be not suitable to gymnosperm SAM, not have stable surface tunica layer. In 1938, American botanist A. Foster described zones in gymnosperm SAM based on the cytohistologic differentiation and thus called it a cytohistological zonation theory. With works by E. Gifford, in 1954, this zonation pattern was demonstrated to be also applicable to angiosperm SAM. As another theory, in 1952 French botanist R. Buvat proposed the m${\acute{e}}$rist${\grave{e}}$me d'attente (waiting meristem) theory: however, this concept was confuted because of its negation of function during vegetative growth phase to central initial cells. Rescent studies with Arabidopsis thaliana have found that formation and maintenance of the SAM are under the control of selected genes: SHOOTMERISTEMLESS (STM) gene forms the SAM, and WUSCHEL (WUS) and CLAVATA (CLV) genes function in maintaining the SAM; signaling between WUS and CLV genes act through a negative feedback loop.
Kim, Youn Hee;Lee, Gee Young;Kim, Hye Hyeong;Lee, Jae Hong;Jung, Jae Hong;Lee, Sang Deok
Journal of Plant Biotechnology
/
v.46
no.1
/
pp.22-31
/
2019
The objective of this study was to investigate the suitable parts for callus induction and optimal concentrations of growth regulators contained in the medium affecting shooting and rooting Echeveria laui and Echeveria elegans for in vitro mass production. To determine the suitable plant parts for callus induction, the leaves were divided into upper, medium and bottom parts and cultured on MS medium at different concentrations with $0{\sim}2mgL^{-1}\;NAA$ and $0{\sim}4 mgL^{-1}BA$. The upper and middle parts of leaves both showed 100% callus formation rate with $NAA\;1\;mgL^{-1}$ and $BA\;1\;mgL^{-1}$ treatment in E. laui. The middle parts of leaves showed 83.3% callus formation rate at $NAA\;2\;mgL^{-1}$ and BA 4 mgL-1 treatment in E. elegans. The shoot induction rate from callus was highest at $NAA\;0.1\;mgL^{-1}$ and $BA\;3\;mgL^{-1}$ treatment in E. laui and $NAA\;0.3\;mgL^{-1}$ in E. elegans. In addition, the number of shoots formation was 10.4 shoots high in $NAA\;1\;mgL^{-1}$ and $BA\;1\;mgL^{-1}$ treatment in E. laui and 12.0 shoots in most effective $NAA\;1\;mgL^{-1}$ and $BA\;0.1\;mgL^{-1}$ treatment in E. elegans. In the case of acclimatization of regenerated plant, growth characteristics did not show any significant difference (35 ~ 55%) shading with respect to the different ratio of substrate mixture, and it was determined that would be appropriate considered plant height and appearance preference of E. laui and E. elegans. It was established that the optimization of culture condition was responsible for the mass propagation in vitro cultures of E. laui and E. elegans.
The explants of yacon (Polymnia sonchifolia Poeppig & Endlicher) were cultured to invest th8e dedifferentiation condition, and formative callus from leaf was cultured to find the regeneration and micro-crown bud formation. Basal MS medium was more effective to form callus than 1/2 MS and B$_{5}$ medium. Calli formations from leaf, petiole and lateral bud were more effective on MS medium supplemented with 1.0, 2.0 mg/L 2,4-D and 0.2, 0.4 mg/L kinetin or BA than 1.0, 2.0 mg/L NAA and 0.2, 0.4 mg/L kinetin or BA. Formative callus from leaf was proliferated about 70% on medium supplemented with 1.0 mg/L BA. When callus was proliferated, 63% regeneration rate was shown on medium supplemented with 1.0, 2.0 mg/L BA in case of subculture for 3~4 months but was not shown on medium supplemented with 1.0, 2.0 mg/L kinetin. Micro-crown bud formed as addition of BA at 3~4 months after callus culture and then was obtained many at 5~6 months, it was most formed about 82% on medium supplemented with 5 mg/L BA. Rate of micro-crown bud formation was increased as more over 5 mg/L BA concentration, when this time, however, shoot had thick leaves and short internodes, and then withered before long, Micro-crown bud was formed about 88.0% on medium supplemented with 5% sucrose, that was more increased 28% than with 3% sucrose. The buds of crown bud between harvested in field and formed in vitro were difference only in size, but both were similar in shape according to histological view.
In order to identify the effect of cadmium uptake by soil ameliorators in rice plants, compost, lime, silicate, and phosphate were treated. Plants were grown with irrigation water containing $0.01mg\;l^{-1}$ cadmium and treated with two irrigation systems, intermittent irrigation and continuous submersion. Soil pH was higher at the panicle formation stage compared to other growing stages. However, Eh value was decreased as growing stage was proceeded. Eh values measured at the continuous submersion plots were lower than those at the intermittent irrigation plots. Cadmium contents in the both leaves and roots of rice plants grown at the continuous submersion plots were lower than intermittent irrigation plots in the middle of growing season. Among the continuous submersion plots, cadmium uptake was highly reduced at the both silicate and lime treated plot compared to other treatments. In the harvest season, cadmium contents in the both leaves and brown rice were higher at the intermittent irrigation plots than the continuous submersion plots. Cadmium uptake was highly reduced at the compost and lime mixture plot compared to other treatments among the continuous submersion plots. Soil pH was negatively correlated with the cadmium contents of the both shoot and brown rice while Eh was positively correlated with those. The cadmium content of shoot was positively correlated with that of brown rice in the harvest season.
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