• Journal of Plant Biotechnology
• /
• v.37 no.4
• /
• pp.494-504
• /
• 2010

The effect of the addition of the fresh medium, volume of under solid medium in double layer culture as well as the medium change after pretreating microspores on the production of embryos in microspore culture of hot pepper (Capsicum annuum L.) has been studied. When cultured after heat pre-treatment, changing pretreatment media with fresh culture media proved to be more effective for embryo production rather than supplementing additional culture media. Heat-pretreating for 3 days turned out more effective for embryo production than pretreating for 1 or 2 days. In the case of anther pretreatment, the addition of fresh medium after culture was not effective for embryo production. In pretreating microspores, however, supplementing additional fresh culture media greatly improved embryo yield and quality. The best time point of media addition was 4 days after culture commenced, and the most effective number of times of media addition was one time addition. Moreover, the effective volume of added medium in double layer culture for embryo production was 1.5 ml. The addition of media more than 1.5 ml reduced both embryo yield and quality. Double layer medium was more effective for embryo development than liquid medium. When the volume of under solid medium increased ranging from 3 ml to 7 ml, more cotyledonary embryos were produced in either 5 ml or 7 ml compared to 3 ml, even though the total number of embryos were highest in 3 ml. These results can be used as an important data for establishing an efficient microspore culture system for producing high frequency of normal embryos in hot pepper.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.42 no.3
• /
• pp.306-316
• /
• 1997

Embryogenic calli were induced from mature seed scutella of anther culture-derived rice variety Zhonghua 8. Cell suspension cultures were initiated from friable embryogenic calli and utilized as source material for protoplast isolation. Generally, the older and finer cell suspensions gave higher protoplast yields than younger suspension cultures. Protoplasts exhibited sustained cell division and formed microcalli when cultured in KPR medium supplemented with 0.5 mg $l^{-1}$ 2,4-D, 1.0 mg $l^{-1}$ NAA and 0.5 mg $l^{-1}$ zeatin using the agarose embedding procedure without feeder cells. Protoplast plating efficiencies ranged from 0.20 to 0.54％. Microcalli were transferred to MS medium supplemented with 2.0 mg $l^{-1}$ kinetin and 0.5mg $l^{-1}$ NAA for plant regeneration. The regeneration frequencies were 2 to 12％, depending on the cell suspension lines of Zhonghua 8. The plants were transferred to the glasshouse and were fertile.

• Journal of agriculture & life science
• /
• v.43 no.6
• /
• pp.29-33
• /
• 2009

The purpose of this study was carried out to investigate the effects of plant growth regulators on in vitro propagation of Liliem cernum Komarov. Small bulblets were poliferated from callus explants after 2 weeks and leaf, root and bulb were formed after 4 weeks culture. Leaf differentiation was promoted vigorously by the combination of TDZ 0.1 mg/L and NAA 0.01 mg/L(87.5%). The rate of root differentiation was the greatest at BA 0.2 mg/L alone(81.8%). The rate of callus formation was the high in medium containing TDZ. The number of bulblets and leaves formed in bulb scales was the greatest at TDZ 0.1 mg/L(5.7). Also, the longest length of total length, leaf and root length were in Zeatin 1.0 mg/L + NAA 0.1 mg/L(10.5 cm). However the longest bulblet was in TDZ 0.1 mg/L(1.4 cm).

• Journal of Plant Biotechnology
• /
• v.32 no.1
• /
• pp.37-44
• /
• 2005

Microspores were isolated from pepper (Capsicum annuum L.) anthers by using a micro-blender and cultured in modified NLN medium at $25^{\circ}C$. The influence of pretreatment period at $32^{\circ}C$, adding the 2-hydroxynicotinic acid to a pretreatment medium, and co-pretreatment anthers with microscopes on the induction of embryo were examined. Globular and torpedo embryos were observed from 3 weeks after culture. Embryo development was not synchronized within culture. After 4 weeks in culture, in addition to globular and torpedo embryos, cotyledonary embryos were observed. Normal cotylodonary embryos developed into plantlets when transferred to a solid hormone free B5 medium containing $2\%$ sucrose. Embryo yields were significantly higher after 1- and 2-day pretreatment at $32^{\circ}C$. However the development of embryo ceased at the globular or heart stage. In contrast, embryo yields were lower after 3- to 6-day pretreatment at $32^{\circ}C$ and embryo developed at the cotyledonary stage. After adding the 2-hydroxynicotinic acid to anther pretreatment solution, embryo yields were slightly increased. However most embryos occurred were at the globular or heart stage. Co-pretreatment of microspores with anthers was deleterious for embryo induction and development. AS far as we know, this is the first report of success in obtaining high frequency of embryogenesis and plantlets formation from isolated microspores of pepper. Although the culture conditions have to be optimized further, this promising microspore culture system can be used for genetic transformation, selection for dominant and recessive traits as well as for the production of homozygous doubled haploid plants.

• Current Research on Agriculture and Life Sciences
• /
• v.1
• /
• pp.1-9
• /
• 1983

This study was conducted to obtain basic information on the rice anther culture. Materials used were (Inabawase X YR 2404-14-2-1) $F_1$ hybrid. Callus growth rate on various media, induction frequency of callus in spikelet and panicle, and the effect of treatment on anther and callus were evaluated. The results obtained were summarized as follows ; The growth rate of callus on N-6, M-S, P.E.agar media was 19.8, 13.1, 4.1 times respectively after 30 days inoculated, and on liquid media was 3.8, 5.1, 1.4 times, respectively. Organ differentiation on N-6, M-S, P.E.agar media was 37.5%, 12.5%, 17.5% respectively. The difference of induction frequency of callus per panicle was 0.14%-6.25% and per spikelet was 0-19.05%. Almost callus was induced 30-35 days after inoculation. Organ differentiation of induced callus was decreased by culture. Callus cultured for 13 days after induction did not make shoot. Anthers cold shocked at $8^{\circ}C$ for 5 days obtained 3.32% efficiencys of callus induction per number of anthers plated, and compared with 2.41% of no treated anthers. But anther treated at $8^{\circ}C$ for 7 days decreased 2.24%. Callus induction periods were shortened by cold treatment for about 5 days. Callus cultured on medium containing 2 mg/l of 2, 4-D showed 5% on root formation but medium containing 5 mg/l of 2, 4-D showed 30% of root formation after transfered on the medium without 2, 4-D. Callus cold shocked at $15-18^{\circ}C$ revealed poor efficiency for root formation, but 5 days treatment was good for shoot formation.

• Journal of Plant Biotechnology
• /
• v.43 no.1
• /
• pp.76-81
• /
• 2016

We performed a molecular marker-based analysis of quantitative trait loci for traits that determine the quality of appearance of grains using 120 doubled haploid lines developed by anther culture from the F1 cross between 'Cheongcheong' (Oryza sativa L. ssp. Indica) and 'Nagdong' (Oryza sativa L. ssp. Japonica). We therefore calculated the alkali digestion value (ADV), used to indirectly measure gelatinization temperature, to evaluate the quality of cooked rice in 2013 and 2014. The ADV score of frequency distribution was higher milled rice than brown rice. In total, nine different quantitative trait loci (QTLs) were found on 5 chromosomes in 2013 and 2014. Also, chromosome 5, 8 were detected over two years. We conclude that selected molecular markers from this QTL analysis could be exploited in future rice quality. In conclusion, we investigated ADV of brown and milled rice in CNDH population. This study found nine QTLs related to the ADV of brown and milled rice. The detected one marker can be used to select lines with desirable eating-quality traits because ADV is closely associated with the eating quality of cooked rice. Therefore, it will be useful to collect resources and distinguishable in many varieties for rice breeding program.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.46 no.4
• /
• pp.261-265
• /
• 2001

This study was carried out to select randomly amplified polymorphic DNA (RAPD) markers associated with grain weight of a large-grain mutant, Hyacp 39-26-1, derived from anther culture of a rice cultivar, 'Hwayeongbyeo'. The segregation mode for grain weight in an F$_2$ population from a cross, 'Hwayeongbyeo/Hyacp 39-26-1', showed a nearly normal distribution. One hundred and ninety-one F$_2$plants ranged from 21.8 g to 34.7 g in 1,000-grain weight with a mean of 26.8 g. Five hundred and twenty primers were used to detect the RAPD markers associated with the grain weight of the large-grain mutant. Of these primers, 54 primers showed polymorphism between 'Hwayeongbyeo' and 'Hyacp 39-26-1'. Four RAPD markers (OPB18, OPH07, OPT20, and OPX20) were significantly related to the grain weight of twenty one F$_3$ lines derived from the cross, 'Hwayeongbyeo/Hyacp 39-26-1'. This RAPD marker could facilitate the early and efficient selection of high-yield lines through improvement of grain weight in rice.

• Journal of Plant Biotechnology
• /
• v.29 no.3
• /
• pp.199-207
• /
• 2002

The tropane alkaloids hyoscyamine (its racemic form being atropine) and scopolamine are used medicinally as anticholinergic agents that act on the parasympathetic nerve system. Because they differ in their actions on the central nervous system, currently there is a 10-fold higher commercial demand for scopolamine, in the N-butylbromide form, than there is for hyoscyamine and atropine combined. Several solanaceous species have been used as the commercial sources of these alkaloids, but the scopolamine contents in these plants often are much lower than those of hyoscyamine. For this reason there has been long-standing interest in increasing the scopolamine contents of cultivated medicinal plants. Naturally occurring and artificial interspecific hybrids of Duboisia have high scopolamine contents and are cultivated as a commercial source of scopolamine in Australia and other countries. Anther culture combined with conventional interspecific hybridization also has been used to breed high scopolamine-containing plants in the genera Datura and Hyoscyamus, but without much success. The use of recombinant DNA technology for the manipulation of metabolic processes in cells promises to provide important contributions to basic science, agriculture, and medicine. In this review, I introduce on the enzymes and genes involved in tropane alkaloid biosynthesis and current progress in metabolic engineering approaches for tropane alkaloid, especially scopolamine, production.

• Horticultural Science & Technology
• /
• v.29 no.5
• /
• pp.488-493
• /
• 2011

Callus culture initiation of strawberry (Fragaria${\times}$ananassa Duch.) was investigated at different Murashige and Skoog (MS) medium strengths, types and concentrations of plant growth regulators, and incorporating a cold pretreatment period to determine the optimal nutritional and environmental conditions. No high quality callus was induced on MS media without auxin regardless of medium strength. When 6-benzylaminopurine (BA) was combined with indole acetic acid (IAA), naphthalene acetic acid (NAA), and 2,4-dichlorophenoxyacetic acid (2,4-D), high quality callus were highly induced compared to medium supplemented with auxin alone. When $0.5mg{\cdot}L^{-1}$ BA was combined with IAA, NAA, and 2,4-D, high quality callus induction was more effective than the medium supplemented with the other BA concentrations. The best combination of auxin and cytokinin for high quality callus induction was $1.0mg{\cdot}L^{-1}$ NAA and $0.5mg{\cdot}L^{-1}$ BA. Although the differences in callus induction were not significant, high quality callus induction at half strength MS medium was more effective than at full strength medium. When $30g{\cdot}L^{-1}$ sucrose was added to the half strength MS medium, the rate of high quality callus induction increased. The optimum cold pretreatment temperature and period for high quality callus induction were $4^{\circ}C$ and 72 h, respectively. Regeneration rate of high quality callus increased in MS medium supplemented with thidiazuron.

• Korean Journal of Plant Tissue Culture
• /
• v.21 no.6
• /
• pp.333-339
• /
• 1994

Enzyme activities and phenolic compound were compared to investigate the physiological characteristics during shoot formation from diploid and haploid of Nicotiana tabacum w BY4. The Nakata medium with 1.0 mg/L IAA, 0.5 mg/L Kinetin and 3 g/L activie carbon was excellent to induce the haploid plants from the middle size anther within 30 days after culture. The MS medium with 0.5 mg/L 2,4-D was good for callus induction from leaf explants of diploid and haploid, and a lot of plane were regenerated from calli on the MS medium supplemented with 2.0 mg/L BAE Activities of peroxidase for both of diploid and haploid plane were the highest at 2.0mg/L BAP Activities of IAA oxidase and catalase of haploid Plants were high or than those of diploid plants. On the other hand, activity of peroxidase of haploid plants were lower than those of diploid plants.