• Horticultural Science & Technology
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• v.29 no.5
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• pp.494-499
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• 2011

Cold pretreatment, washing medium and composition of nutrient media may have marked effects on microspore embryogenesis. When microspores isolated from radish (Raphanus sativus L. cv. Gwanhun) flower buds were washed with Nitsch & Nitsch (NLN) medium liquid medium containing $130g{\cdot}L^{-1}$ sucrose (NLN-13), yields of microspore-derived embryos were greater than when using B5 liquid medium containing $130g{\cdot}L^{-1}$ sucrose. Microspore viability is known to decrease rapidly with storage; however, in this experiment, microspore viability was maintained for 24 h at $4^{\circ}C$ without media. Among the various medium concentrations used ($0.25{\times}$, $0.5{\times}$, $1.0{\times}$, $2.0{\times}$, and $4.0{\times}$ NLN liquid medium), $0.5{\times}$ NLN liquid medium induced the most efficient formation of microspore-derived embryos. In addition, microspore-derived embryos yields were greater when microspores were cultured in $0.5{\times}$ NLN liquid medium supplemented with $0.25{\times}$, $0.5{\times}$, and $1.0{\times}$ NLN microelements, compared to medium not supplemented with microelements. In this study, the highest yield of microspore-derived embryos was observed when the microspores derived from flower buds were washed using NLN-13 liquid medium and then cultured on $0.5{\times}$ NLN liquid medium supplemented with $0.25{\times}$ NLN microelements, followed by incubation at $25^{\circ}C$ for 30 days.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.51 no.spc1
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• pp.237-241
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• 2006

This experiment was carried out comparison with haploid plants productivity by microspore culture among domestic cultivars of Brassica napus L. Isolated microspore from flower buds were cultured on NLN medium supplemented with 13% sucrose, $0.05mg/{\ell}$ BA and $0.5mg/{\ell}$ NAA. Genotype was important factor in haploid embryo productivity 'Tamlayuchae' showed the highest haploid embryo production frequency (176 embryos formed from 1 flower bud). But, 'Hallayuchae' and 'Youngsanyuchae' were not generated embryo even cell division. When suspension culture on NLN liquid medium at 100 rpm, embryos were developed multilobe abnormal embryo cluster. Multilobe abnormal embryos on MS medium basal solid medium were regenerated multiple shoots. Regenerated haploid plant with well developed shoots and roots on MS basal medium were successfully transferred to pots.

• Korean Journal of Plant Resources
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• v.37 no.3
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• pp.225-234
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• 2024

Chinese lobelia (Lobelia chinensis Lour.) is an important medicinal plant that is used in traditional Chinese, Korean, and Japanese medicine. The goal of the current study was to develop an in vitro propagation technique for Lobelia chinensis. We have examined the effects of different media compositions on the regeneration of shoots from nodal cultures of Lobelia chinensis, including Murashige and Skoog (MS), Gamborg (B5), Schenk and Hildebrandt (SH), Woody plant (WPM), Chu (N6), and Nitsch and Nitsch (NLN) media. Similar to this, shoot regeneration was examined using MS medium of double (2.0), full (1.0), half (0.5), and quarter (0.25) strengths. The regeneration of shoots was also examined with additions of 0, 1, 3, 5, and 7% (w/v) sucrose to MS media. For axillary shoot regeneration, full-strength MS medium supplemented with 3% (w/v) sucrose was shown to be the most effective of all the evaluated factors. On this medium, nodal explants optimally regenerated 4.5 shoots per explant and subsequently shoots involved in rooting on the same medium. The regenerated plants possessed abundant phenolics, flavonoids, and DPPH, ABTS, and FRAP antioxidant activities. High performance liquid chromatographic examination (HPLC) of the regenerated plants revealed an accumulation of myricetin and catechin in higher amounts.

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

• Journal of Plant Biotechnology
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• v.44 no.1
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• pp.27-34
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• 2017

Our aim was to find the effective duration and culture conditions for microspore culture in a genetically variant cabbage (Brassica oleraceae L. var. capitata). We discovered that the '$2{\times}NLN$ medium containing $AgNO_3$ 1 mg/l, sucrose 13%' was most efficient in producing the cabbage embryos. The number of induced embryos was higher in $F_2$ or $F_3$ progeny generation than the $F_1$ hybrid cultivar used as plant materials. Thus, we recommend microspore culturing using progeny plants of failed $F_1$ cultivar. The acquisition ratio of embryos and plants derived from microspore was higher in the early flowering period as compared to the late flowering period. When transplanted in the soil, 71.2% plants developed from the early flowering period, compared to 27.0% from the middle period and 1.8% plants from the late period. Thus, we recommend microspore culture using buds collected during the early flowering period.

• Horticultural Science & Technology
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• v.30 no.5
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• pp.573-578
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• 2012

A total of eleven Chinese cabbage accessions were used for microspore culture and were grown to take basal data. Based on the collected data, breeding materials were chosen to develop new improved Chinese cabbage cultivars. The range of microspore-derived embryoid taken from flower buds was 1.6 to 35.4 embryoids. The embryoids from IT26110 and IT26153 among the Chinese cabbages were more than 34 per flower bud. The viability rate after cold treatment was low from 0.2 to 11.7%. The range of fertility rate was 7.7 to 58.8% in general but the IT26118, IT26122, IT26128, IT26130, and IT26164 were more than 50%. The result of their seed production ability by selfing was 11.9 seeds per siliqua in IT26128 while the others were less than 10 seeds. In the microspore culture using parents of different hereditary, the number of embryoids, the number of plants, the rate of fertility and their pure seed production ability appeared to be very different in doubled haploid lines obtained from fertile plants of Chinese cabbage.