• Title/Summary/Keyword: cotyledon culture

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High Frequency Somatic Embryogenesis and Plant Regeneration from Cultured Immature Seeds of Ostericum koreanum Kitagawa and Angelica purpuraefolia Chung (강활(Ostericum koreanum)과 지리강활(Angelica purpuraefolia)의 미숙종자로부터 고빈도의 체세포배 발생과 식물체 재분화)

  • 최은경;박학봉
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.5
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    • pp.299-306
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    • 1995
  • The objective of this study is to establish an efficient celll culture system for somatic embryogenesis in Ostericum koreanum and Angelica purpuraefolia. The highest frequency of embryogenic callus on immature seeds of O. konanum and A, purpuraefolia. was obtained when seeds were cultured on MS medium containing 0.1 mg/L 2,4-D and 0.1 mg/L BA. However somatic embryos were formed directly from the edge of cotyledon and hypocotyl of plant which regenerated on medium supplemented with 0.1-3.0 mg/L NAA. Immature seed explane cultured at 25$\pm$2$^{\circ}C$ after 10 days treatment at 5$^{\circ}C$ produced embryogenic callus and somatic embryos, and these differentiated into whole plane. Addition of glutamine and coconut milk to media did not enhance the frequency of somatic embryogenesis in immature seed cultures of A. purpuraefolia. However in immature seed culture of O. koreanum, the frequency of somatic embryogenesis were increased on media supplemented with glutamine and 10% coconut milk. Especially addition of glutamine to the medium substituted effect of NH$_4$N0$_3$ in constast to coconut milk. The highest frequency of conversion somatic embryos into plantlet was 89.1% on MS basal medium Embryogenic calli were grown vigorously when maintained on medium with 0.01 mg/L 2,4-D and 0.01 mg/L BA.

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Somatic Embryogenesis and Plant Regeneration from Immature Zygotic Embryo Culture in Pepper (Capsicum annuum L.)

  • Jo, Jeong-Yon;Choi, Eun-Young;Choi, Dong-Su;Lee, Kwang-Woong
    • Journal of Plant Biology
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    • v.39 no.2
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    • pp.127-135
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    • 1996
  • An efficient system of somatic embryogenesis was established for the red pepper plant (Capsicum annuum L. cv. Nokkwang) usign immature zygotic embryos. The size of the immature zygotic embryos and the concentrations of 2, 4-D and sucrose were found to be critical. Somatic embryos were induced via callus or directly from explants and regenerated into plantlets successfully. When zygotic embryos 1~2 mm long were cultured on the modified Murashige-Skoong (MS) medium supplemented with 2 mg/L 2, 4-D for 3 weeks in the dark, somatic embryos were induced directly from the apical region of zygotic embryos with the highest frequency being approximately 90%. To mature the somatic embryos, ABA and an ethylene inhibitor AgNO3 were used. The highest frequency of shoot regeneration (25% in each) resulted at 2$\mu$M ABA or 20$\mu$M AgNO3 treatment at rates 3.7 and 1.6 times control, respectively. Shoots developed mainly from the cotyledonary node on CoCl2-containing medium, and from the upper side of cotyledon on medium containing AgNO3 while the embryos on the control medium produced shoots from both the cotyledonary node and the upper region of cotyledons both at frequencies of 50%. Indirect somatic embryogenesis via callus was induced at an efficiency of approximately 10% with zygotic embryos 3~4 mm long cultured on MS medium containing 5~10 mg/L, 2, 4-D for 5~7 weeks under a continuous light condition. The plants regenerated from the somatic embryos were morphologically normal. Using scanning electron microscopy, the direct and indirect somatic embryogeneses were observed to follow the globular, heart and torpedo stages, similar to zygotic embryogenesis. Also, suspensors appeared in the early globular and ovoid-shaped late globular embryos during indirect somatic embryogenesis.

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High Frequency Shoot Formation and Plant Regeneration from Cotyledonary Hypocotyl Explants of Boxthorn (Lycium chinense Mill.) Seedlings (구기자나무(Lycium chinense Mill.) 유식물체의 자엽하배축으로부터 고효율의 부정아 형성과 식물체 재생)

  • Jo, Man-Hyun;Ham, In-Ki;Lee, Bong-Chun;Kim, Jae-Whune;Lee, Won-Seok;Kwon, Suk-Yoon;Lee, Haeng-Soon;Kwak, Sang-Soo
    • Journal of Plant Biotechnology
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    • v.31 no.3
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    • pp.203-207
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    • 2004
  • To establish high frequency shoot formation from two cultivars (Cheongyangjaerae and Myungan) of boxthorn (Lycium chinense Mill.), hypocotyl segments with cotyledons from seedlings were used as explants. High frequency adventitious shoot formation (more than 80%) were obtained from hypocotyl segments with cotyledon on MS medium supplemented with 1.0 mg/L zeatin, when precultured for 3 weeks under dark conditions followed by transfer to light conditions. But there was no shoot induction in the explants cultured without preculture under dark conditions. Roots were induced from the shoots when transferred to rooting medium supplemented with 1.0 mg/L IAA for 4 weeks. Regenerated plantlets were grown to normal mature plants in soil.

Changes of Nitrogen Compounds and Nutritional Evaluation of Soybean Sprout - Part III. Changes of free amino acid composition - (콩나물 제조중(製造中) 질소화합물(窒素化合物)의 변화(變化)와 그 영양학적(營養學的) 연구(硏究) - 제3보(第三報). 유리(遊離)아미노산의 조성변화(組成變化) -)

  • Yang, Cha-Bun
    • Applied Biological Chemistry
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    • v.24 no.2
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    • pp.101-104
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    • 1981
  • Changes in free amino acid composition in cotyledon and axis was investigated during growth of soybean sprout. Free amino acid increased with sprout-growth, especially in axis. Arginine, proline and methionine did not appear in axis throughout. Free amino acids in total amino acids was 0.11% for soybean and 8.8% for 8 day-sprout. Free amino acid change in the course of culture was in order of Asp>His>Val>Ile>Thr>Ser>Lys>Tyr>Phe>Leu>Ala>Gly>Pro>Arg>Glu>Met. The content of free amino acid in 4 day-sprout was in order of Asp>Ser>Val>His>Ala>Ile>Lys>Thr>Arg>Leu>Glu>Phe>Tyr>Gly>Cys>Met>Pro.

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Plant Regeneration via Somatic Embryogenesis and Organogenesis from Peucedanum japonicum $T_HUNB$ (갯기름나물의 체세포배 분화와 기관 분화를 통한 재분화)

  • Kim, Ok-Tae;Kim, Kwang-Soo;An, Jun-Chul;Hwang, Baik
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.1
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    • pp.21-24
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    • 2001
  • Peucedanum japonicum $T_{HUNB}$ used as a edible and medicinal plants was investigated for in uitro regeneration. Callus formation occurred on leaf and stem explant cultures and showed spontaneous embryogenic and organogenic capability on MS basal medium supplemented with 0.1~5 mg/L NAA and 0~10 mg/L BA in dark. The regeneration was highest on the condition supplemented with 2.5 mg/L NAA and 10 mg/L BA. Development of the somatic embryo progressed through the globular, heart-shaped, torpedo-shaped and cotyledonary stage, typical of zygotic embryos. When the first somatic embryos was cultured on the medium supplemented with 0.2 mg/L NAA, secondary somatic embryo were induced with higher frequency on the hypocotyl then on the cotyledon and root.t.

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Analysis of Dietary Fiber of 66 Korean Varieties of Sprout Beans and Bean Sprouts

  • Lee, Kyung-A;Cho, Young-Ae;Hwang, Young-Hyun;Lee, Hye-Sung
    • Preventive Nutrition and Food Science
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    • v.8 no.2
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    • pp.173-178
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    • 2003
  • The present study was conducted to determine a high-fiber variety of sprout bean and bean sprout. Sixty-six varieties of Korean sprout beans and thirty varieties of Korean bean sprouts were analyzed for their total dietary fiber (TDF) using a combination of enzymatic and gravimetric methods adopted by AOAC. The average and range of the TDF contents of the 66 varieties of sprout beans were 21.73$\pm$2.43% and 16.60~29.2% (dry basis), respectively. The top five types of sprout beans with high TDF anions the 66 samples were KLG10658 (29.2%), Dawonkong (28.46%), Sohokong (25.66%), Moohankong (25.465%) and Samnamkong (24.94%). The mean TDF content of sprout beans was relatively high in the variety with yellow seed coat color and a smaller seed size (<15 g/100 seeds) than the ones with other colors and of medium or large seed size. The average and range of the TDF content of the 30 varieties of bean sprouts were 24.48$\pm$3.9% and 18.3~33.38%, respectively. Dawonkong sprouts showed the highest TDF content (33.38%). The average TDF content of bean sprouts was 1.16 times higher than that of the corresponding seed beans. The TDF content of bean sprouts did not significantly change during the days of culture. The TDF content in the different parts of the bean sprout were high in the order of roots, hypocotyl and cotyledon.

Expression of Chinese Cabbage Glutathione Reductase Gene in Lettuce (Lactuca sativa L.) (형질전환 상추에서 배추 Glutathione Reductase 유전자의 발현)

  • 정재동;김창길;조진기
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.4
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    • pp.267-271
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    • 1998
  • Cotyledon explants of lettuce were cocultured with Agrobacterium tumefaciens LBA4404::pBKS-GR1 harboring glutathione reductase(GR) gene in MS medium supplemented with 0.1 mg/L NAA and 1.0 mg/L 2ip for 48 hr. These explants were transferred to MS medium supplemented with 0.1 mg/L NAA and 1.0 mg/L 2ip, 50 mg/L kanamycin, and 500 mg/L carbenicillin. After 4 weeks of subculture, kanamycin-resistant shoots were obtained on selection medium. Leaves of putative transformants survived on selection medium containing 100 mg/L kanamycin. Incoporation of the GR gene into lettuce was confirmed by PCR analysis of genomic DNA. Southern blot analysis showed that ECL-labeled GR gene was hybridized to the expected amplified genomic DNA fragment of about 1.8 kb from transgenic lettuce. The presence of mRNA in transgenic lettuce was confirmed by RT-PCR with total RNA of transgenic lettuce. In progeny test of transformants, R$_1$ seeds were resistant to kanamycin (200mg/L) on MS medium.

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Effect of Abscisic Acid on the Number of Somatic Embryo Cotyledons in Tissue Cultures of Aralia cordata Thunb. (땅두릅(Aralia cordata Thunb.)의 조직배양에서 체세포배의 자엽 수 변화에 미치는 ABA의 영향)

  • 이강섭;소웅영
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.5
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    • pp.287-291
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    • 1994
  • In order to elucidate the effect of abscisic acid (ABA) on the abnormality of somatic embryos, somatic embryos were induced from embryogenic cell clumps derived from cotyledon segment of Aralia cordata. When embryogenic cell clumps were pretreated medium containing 0.2 mg/L ABA for 3 weeks before transferring to MS basal medium, the frequency of embryos with normal cotyledons enhanced 68% as compared with control. However when clumps pretreated in medium containing 0.2 mg/L ABA were transferred to medium containing 0.1 mg/L ABA, the Sequency decreased to about 29%. In the case of globular embryos cultures in medium containing various concentrations of ABA (0.01 to 1.0mg/L), the frequency of dicotyledonary embryo formation decreased propotionally to ABA concentration. Also, when somatic embryos at various stages were cultured in medium containing ABA, those with polycotyledons appeared at higher frequency.

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Transformation of PAT gene into Lettuce (Lactuca sativa L.) using Agrobacterium tumefaciens (Agrobacterium tumefaciens를 이용한 상추 (Lactuca sativa L.)의 PAT유전자 형질전환)

  • 류정아;김창길;이현숙;최경배;양덕춘
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.4
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    • pp.197-200
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    • 2001
  • Agrobacterium tumefaciens MP90 harboring PAT (phosphinothricin acetyltransferase) and NPTII-GUS gene were used for the genetic transformation of lettuce (Lactuca Sativa L.). Shoot regeneration from cotyledon explants were obtained from the MS medium supplemented with 0.1 mg.L$^{-1}$ NAA, 1.0 mg.L$^{-1}$ 2ip, 50 mg.L$^{-1}$ kanamycin and 500 mg.L$^{-1}$ carbenicillin after cocultivation with A. tumefaciens for 2 days. Kanamycin resistance test of transgenic plants indicated that the NPTII gene was integrated into the lettuce genome and was stably expressed. PCR and northern blot analysis indicated that bialaphos resistance gene (PAT) was stably integrated into the lettuce genome. The transgenic plant sprayed with Basta (1500x) remained healthy with continuous growth, while the control group exhibited fatality.

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Esterase isozyme patterns in developing plant regeneration from calli of citrus junos Sieb.

  • Lee, Hyun-Hwa;Lee, Sook-Young;Park, Min-Hee;Jang, Hyun-kyu;Kim, Hong-Sub
    • Plant Resources
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    • v.2 no.1
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    • pp.1-9
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    • 1999
  • The callus from the hypocotyl region of immature embryo of Citrus junos Sieb. was efficiently induced in the $\frac{1}{2}$ MS medium containing 45uM BA after 8 weeks culture. The callus was developed into the two callus type, embryogenic callus and nonembryogenic callus, which can be distinguished by visual examination depending on color and appearence. In vitro regeneration of callus established efficiently in the hormone-free MS medium from the embryogenic callus. In order to investigate the physiological changes depending on the developmental stage of embryo, the embryo was formed in the MS medium. The embryogenic and nonembryogenic callus, and the various stages of the somatic embryo were examimed the changes of esterase activity, and their isozyme patterns as well. The protein content and esterase activities was gradually increased on the developmental stages of embryo. Total protein pattern were different by the SDS-PAGE and were appeared strong band of 23 KD in the torpedo stage. The pattern of the esterase isozyme was exhibited a difference between embryogenic callus and nonembryogenic callus. It was appered pI 6.0, 8.0, 8.2 in the embryogenic callus. Also the new band of pI 4.75 was appeared in the cotyledon. These results suggest that the changes of esterase activities and isozyme patterns are importent factor in the differentiation and development of citrus.

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