• Journal of Plant Biotechnology
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• v.38 no.4
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• pp.258-262
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• 2011

Embryogenic callus was induced from the cultures of immature embryos of Camellia japonica L. on Murashige & Skoog's (MS) solid medium supplemented with 1 mg/L 2,4-dichlorophenoxy acetic acid (2,4-D), and then the embryogenic callus was proliferated on same medium for 4 weeks over. The embryogenic callus was sub-cultured on MS basal medium without 2,4-D to produce coyledonary stage of somatic embryo. The frequency (%) of somatic embryogenesis was 25.1%, and the majority of somatic embryos formed had a abnormal morphology with cupshaped cotyledon (48.3%), one cotyledon (12.6%), three cotyledons (9.4%), four cotyledons (1.9%), whereas was only normal morphology with two cotyledon (27.5%). When the somatic embryos with normal or abnormal cotyledons transfer to MS basal medium or $\frac{1}{2}$ MS medium with/or without plant growth regulators ($GA_3$, IBA) for regeneration, the frequency (%) of two-cotyledon embryos regenerated into plantlets was higher 11.1% than one cotyledon (0.0~8.3 %), three cotyledons (0.0~5.8%), four cotyledons (0.0%), cup-shaped (0.3~4.2%). These results demonstrated that the anomalous cotyledons of somatic embryos could caused to decrease the rate of plant regeneration.

• Journal of Plant Biotechnology
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• v.47 no.2
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• pp.179-184
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• 2020

Embryogenic calli from Codonopsis lanceolata L. were cultured in MS liquid media and supplemented with various concentrations of primary carbon sources to study the effects of carbohydrates and osmoticum on somatic embryogenesis and somatic embryo morphology. Sucrose, glucose, and a combination of 3% sucrose and various concentrations of sorbitol or mannitol as osmoticum were used as carbon supplements. The maximum number of somatic embryos per flask was greater in media exclusively supplemented with 3% sucrose (128.29) than exclusively glucose-supplemented media (47.67) and either supplement combination of 3% sucrose and osmoticum (95.67 with mannitol and 114.00 with sorbitol). The number of somatic embryos gradually decreased in media with increasing concentrations of combined osmoticum supplement. Decreases also occurred in the highest concentrations of sucrose- and glucose-supplemented media. The total frequency of somatic embryos with two cotyledons was slightly higher in medium with 3% + mannitol (24.09%) compared with exclusively sucrose (21.52%), glucose (21.22%), or 3% sucrose + sorbitol (22.13%). As concentrations of sucrose and glucose increased, the occurrence of two cotyledons and trumpet cotyledons gradually decreased and the occurrence of polycotyledon and globular stage embryos increased. Furthermore, as concentrations of 3% sucrose and osmoticum increased, the occurrence of trumpet cotyledon and globular stage embryos increased and the occurrence of polycotyledon gradually decreased. These results demonstrated that the somatic embryogenesis and occurrence of cotyledon morphology were influenced by the concentration of carbohydrates and combinations of 3% sucrose and osmoticum supplements.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.39 no.5
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• pp.480-488
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• 1994

Low seedling growth rates of white clover (Trifolium repens L.) have been limited its good establishment to pastures. The experiment was done to determine the effect of removal of cotyledon and unifoliolate on the growth and morphological characters of contrasting white clover cultivars for 8 weeks after the treatment. Individual plants of cv. Regal (large leaf), Louisiana S-l (medium-large leaf), Grasslands Huia (medium-small leaf) and Aberystwyth S184 (small leaf) were grown in 10cm plastic pot containing a 2:1:1 soil:sand:peat moss mixture until the cotyledon or unifoliolate stage and then removed one (C1) or two cotyledons (C2) at cotyledon stage, and unifoliolate only (U), unifoliolate and one cotyledon (DC1) or unifoliolate and two cotyledons (DC2) at the unifoliolate stage. To measure the removal effect on biomass and morphological characters (leaf area, petiole and stolon lengths, growing tips and leaves), plants were sampled 4, 6 and 8 weeks after the treatment. Intact plants had greater biomass and morphological characters than removal-treated ones, Removal treatments at cotyledon stage, C1 and C2, were decreased more biomass and morphological characters than removal ones at unifoliolate stage while the severer cotyledon removal, the more reduction. Stolon length per plant and petiole length markedly inclined 6 weeks after the treatments although biomass and the other characters continuously did. Relatively large-leaved cultivar had more biomass, leaf area per plant and longer petiole than the other(s) but the reverse results were true in stolon length, growing tips and no. of leaves per plant. Biomass was linearly increased with increased leaf area but the earlier and severer removal, the less slope. The severer damage of cotyledon and unifoliolate had detrimental effects on the growth and aftermath establishment of white clover

• Journal of Plant Biotechnology
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• v.50
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• pp.89-95
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• 2023

In order to investigate the effect of carbon sources on somatic embryogenesis and cotyledon number variations in carrot, embryogenic callus were cultured in the medium supplemented with various concentrations of sucroseor glucose, and with combination of 2% sucrose and various concentrations of mannitol or sorbitol. The maximum number of somatic embryos formed per flask (1,555.70) was obtained in the medium supplemented with 2% sucrose rather than glucose alone or a combination of mannitol or sorbitol and 2% sucrose, and the number of somatic embryos was decreased with the increasing of mannitol or sorbitol concentration. The frequencies of somatic embryos with two cotyledons were 35.14% for sucrose, 19.88% for glucose, 32.55% for mannitol + 2% sucrose, and 38.59% for sorbitol + 2% sucrose, respectively, and the frequencies of abnormal somatic embryos having 3 or more cotyledons were 64.86% for sucrose, 80.12% for glucose, 67.44% for mannitol + 2% sucrose, and 61.41% for sorbitol + 2% sucrose, respectively. Particularly, the frequency of somatic embryos with two cotyledons (59.16%) was the highest in the 2% sucrose medium compared to the frequency of abnormal somatic embryogenesis with three or more cotyledons, and the frequency gradually decreased with increasing concentration of glucose, mannitol or sorbitol. According to these results, it was found that the ratio of abnormal somatic embryo was higher than the normal somatic embryo in carrot, and was shown that somatic embryogenesis and the cotyledon number was affected by the concentrations of sucrose, glucose as carbon source, and mannitol and sorbitol as osmotic agents in culture medium.

• Journal of Life Science
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• v.11 no.2
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• pp.166-172
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• 2001

Various types of the seeding of bottle gourd widely used as a rootstock of watermelon has been required to satisfy the farmers need. The study was done to determine the effect of light quality of blue, red and far-red lights treated with light emitting diodes on growth and morphology of bottle goured seeding and chlorophyll content of its cotyledons. The lights were treated in the growth chamber for 7 days to the seeding elapsed 8 days after sowing under natural condition, and 64 hole trays with commercial bedsoil. Plant height, length and diameter of hypocotyl, leaf area of cotyledon and first true leaf, its leaf length, number of true leaves, fraction and total dry weight were measured. Red light shortened and slenderized the hypocotyl, which lengthened by far-red light and thickened by blue light. Plant height was declined in order of far-red light treatment, blue and red lights mainly due to difference of hypocotyl length, Area and length of the first true leaf became smaller and shorter under far-red light than under the other lights. However, blue light increased leaf area of cotyledons. Two cultivars cv. Yongjadaemok and cv. Kunghap had different response to the light treatments in total seedling dry weight(dw); far-red and red light treatments showed the greatest and the least dw of hypocotyl, respectively, while blue and red lights did the greatest dw of the other organs. Among the ratio of each organ dw to total dw, those of hypocotyl and true leaves were different between the light treatments; the highest ratio of hypocotyl dw to total dw was observed in far-red light treatment but the lowest was in red light treatment. Those of the true leaves were the lowest in far-red light and similar response in blue or red light treatment. Chlorophyll content of cotyledons was decreased in order of red light treatment, blue and far-red lights, meaning that short period light treatment may influence photosynthesis of seeding and afterward its growth.

• Korean Journal of Plant Tissue Culture
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• v.21 no.1
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• pp.7-14
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• 1994

Immature zygotic embryos of five Korean soybean cultivars cultured on Murashige and Skoog's (MS) medium supplemented with various concentrations of 2,4-Dichlorophenoxyacetic acid (2,4-D) produced somatic embryos without forming an intervening callus. The highest frequency (up to 90%) of somatic embryo formation was obtained when zygotic embryos were cultlued on medium containing 1 to 2 mga 2, 0-D in four cultivars. However the frequency was highly variable to the cultivars. Transversely sliced primary somatic embryo halves were also capable of forming secondary embryos at frequencies of up to 70% when cultured on medium containing 0.1 to 1 mg/L 2,4-D. Somatic embryos formed on zygotic embryos cultured on medium containing 0.1 to 0.2 mg/L 2,4-D had two cotyledons more frequently than one horn-type cotyledon and those on medium containing 0.5 to 4mg/L 2,4-LD had a horm-type cotyledon at a prominently higher freequency. However somatic embryos on medium containing 10mg/L or higher concentrations of 2,4-D were usually shunted at the globular stage even after transfer to medium containing lower concentrations of 2,4-D or other growth regulators. non somatic embryos with one or two cotyledons or a hem-type cotyledon were transferred to medium containing $GA_3$, those with two cotyledons converted to plantlets at a higher frequency (25%) than the others.

• Journal of Life Science
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• v.12 no.1
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• pp.19-21
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• 2002

The pathway of embryos formed anther culture in herbaceous peony was influenced by addition of 2,4-D. MS medium with 2,4-Dichlorophenoxy acetic acid (2,4-D) alone did not arise direct embryogenesis, but was proliferate callus. Embryos through calli were produced on medium containing 0.2 mg/1 zeatin or without growth regulators. Direct embryogenesis was obtained from MS basal medium. However, after the anthers were cultured on medium with 0.1 mg/1 2,4-D, 3 g/1 AC, 30 g/1 sucrose, 2 g/1 gelrite for 40 days. Its efficiency (32.3 %) was markedly improved when anthers cultured on medium without 2,4-D. Embryo morphology was also affected by the 2,4-D used in medium. The induction of normal embryos with two cotyledons was higher in the embryos formed through direct embryogenesis than those formed callus. The embryos formed from calli were mainly showed abnormal embryo with one, three, four cotyledons or hors and bowling pin type.

• Korean Journal of Weed Science
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• v.30 no.1
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• pp.1-16
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• 2010

This study was conducted to develope identification method with major broad leaved weed seedlings. Characteristics of weed seedlings were investigated at two to four true leaf stage. Qualitative characteristics were more useful to identification of weed species than quantitative characteristics at the seedling stage. Many species were elliptic and broadly elliptic type in their cotyledon shape. Cotyledon shape of Cassia nomame and Calystegia sepium were broadly oblong. Four species were transverse broadly elliptic in their cotyledon shape. Rumex japonicus, Persicaria hydropiper, Euphorbia supina and Acalypha australis have three or more types in their cotyledon shape. Cotyledons of Vigna angularis var. nipponensis remained below ground after its germination. Many species were round in apex and were cuneate in base of cotyledon. Humulus japonicus and Polygonum aviculare were sessile in cotyledon base. The second true leaf shape of some species changed after first leaf stages. Glycine soja, V. angularis var. nipponensis and Kummerowia striata were broadly ovate, heart-shaped, orbicular or broadly obovate in their first true leaf, respectively, but the second true leaf shape of them were tripalmately compound. Aeschynomene indica and Cassia nomame were paripinnately compound leaf in their first and second true leaves. Margin of true leaves were entire in many species but six species including Xanthium occidentale were dentate in their true leaf margins. Margin of Euphorbia supina was entire in first true leaf and dentate in second true leaf. Margin of Chenopodium album and C. serotinum were entire in their first true leaf stage and erose in their second true leaf stage. The key for weed seedling identification of major weed species were made based on quantitative and qualitative morphological characteristics.

• Plant Biotechnology Reports
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• v.4 no.1
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• pp.53-59
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• 2010

A specific deleted version of ARABIDOPSIS RESPONSE REGULATOR1 (ARR1) lacking the signal receiver domain (1.152 amino acids)-coding sequence, referred to as $ARR1{\Delta}DDK$, was amplified using Arabidopsis thaliana cDNA prepared from adult leaves and transferred into the genome of Nicotiana tabacum cv. Samsun under the transcriptional control of a ${\beta}$-estradiol-inducible expression system. The ectopic expression of $ARR1{\Delta}DDK$ affected the morphology of transgenic seedlings and their segments in vitro. In the presence of an inducer, ${\beta}$-estradiol, ectopic expression of $ARR1{\Delta}DDK$ induced only the formation of soft, pseudo-bulbous tissue in the root tip region of intact seedlings, which appeared similar to callus generated on a hypocotyl segment in the presence of 2,4-D and 6-benzyladenine (BA), both at $1\;{\mu}M$. Those callus tissues on the root tip region could not generate shoots unless $1\;{\mu}M$ BA was supplied. In segment culture, ectopic expression of $ARR1{\Delta}DDK$ induced calluslike tissue around the cut-end of cotyledon and hypocotyl segments with occasional shoot formation, suggesting that the expression of $ARR1{\Delta}DDK$ could substitute for the effects of cytokinin on these segments. Additionally, treatment with only ${\beta}$-estradiol induced NtWUS, a WUS ortholog in tobacco, which was detected during the process of callus tissue formation in the root tip region and also in cotyledon or hypocotyl segments. These findings suggest that the NtWUS might be associated in the transdifferentiation process caused by the functional regulation of $ARR1{\Delta}DDK$ in transgenic tobacco seedlings.

• Horticultural Science & Technology
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• v.35 no.4
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• pp.402-409
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• 2017

We performed seed germination tests to investigate the effects of seed sowing orientation on germination viability on peanut (Arachis hypogaea L.) sprouts. Specifically, we assessed the influence of seed sowing orientation on germination rate, seedling weight, and seedling length, as well as the seedling vigor index. The seeds were sown in oak tree sawdust at 3.0 cm depth. Four seed orientations were tested: vertical with the hypocotyl end down, vertical with the hypocotyl end up, horizontal with the hypocotyl end down, and horizontal with the hypocotyl end up. The mean seed germination percentages of the four seed orientations were significantly different (p < 0.01) and ranged from 25 to 91.7%. The vertical orientation with hypocotyl-end-down and hypocotyl-end-up orientations showed the highest (91.7%) and lowest (25%) germination rates, respectively. The vertical orientation with the hypocotyl end down produced the heaviest (4.9 g) seedlings and the longest hypocotyls (4.65 cm). This orientation also produced the longest true leaf + epycotyl (2.15 cm) and had the highest seedling vigor index (197.1). The seedlings had a straight growth pattern, whereas seedlings from seeds sown with the hypocotyl up had an awkward plumular hook shape. Taken together, to produce peanut sprouts, we recommend placing the seeds vertically with the hypocotyl end down because this orientation leads to a high germination rate, high biomass production, and high overall seedling quality.