• Title/Summary/Keyword: embryo germination

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Effect of Growth Regulators and Osmoticums on Somatic Embryogenesis and Plants Regeneration in Aralia elata Cultivar 'Zaoh' (두릅나무 '자오'의 체세포배 유도와 식물체 형성에 미치는 생장조절제 및 삼투압제 효과)

  • Kim Ji-Ah;Moon Heung-Kyu;Kim Yong-Wook
    • Journal of Plant Biotechnology
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    • v.32 no.2
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    • pp.129-134
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    • 2005
  • Effective micropropagation system via somatic embryognesis was established for a Phytophthora resistant Aralia elata cultivar. Different kinds of growth regulators were needed to induce embryogenic callus with different explant sources. When leaf explants were used, a combination of 2,4-D, TDZ and L-glutamine was needed, whereas when petiole and root explants needed only 1.0 mg/L 2,4-D. Embryogenic callus induction rate under the optimum culture condition was 75.0%, 67.0% and 83.0% from leaf, petiole and root segment, respectively. Somatic embryo germination and plantlet conversion rate appeared to be influenced greatly by various osmoticums. More than 90% of embryos germinated when treated with sucrose, glucose and maltose. However, the highest conversion rate (72%) was recorded on medium with 2% sucrose only. The converted plantlets grew normally on 1/2MS basal medium, were acclimatized on artificial soil mixture and survived more than 95% in the greenhouse condition. The results suggest that the species can be clonally propagated through in vitro culture system via somatic embryogenesis.

Callus growth and plant regeneration from hybrid embryo of L. longiflorum X L. elegans (L. longiflorum X L. elegans의 잡종 배로부터 캘러스의 증식 및 식물체 재분화)

  • Yoon, Eui-Soo;Kwon, Hye-Kyoung;Cho, Yi-Yun
    • Journal of Plant Biotechnology
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    • v.33 no.2
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    • pp.99-104
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    • 2006
  • This study was performed to investigate regeneration of plants differentiated from hybrid embryos between L. longifilorum Georgia and L. elegans Kakutanohikari. In addition, proliferation of callus and process of differentiation were investigated by histological observation. The germination of hybrid embryos was observed in 86 individuals from 48 slice cultures. Plant regeneration was effective on a medium supplemented with 1 mg/L HPh, and only callus proliferation was the highest in combination of 0.1 mg/L HPh and 1 mg/L BA. Also, plant regeneration was the most effective on a medium supplemented with 50 mg/L pyridoxine. We concluded that somatic embryos were formed from procambium of callus and proliferation of embryonic or proembryonic cells were stimulated with NAA from procambial cells.

Fusarium spp. Isolated from Seed, Root and Cultivated Soil of Phaseouls vidissimus and Their Pathogenicity (녹두종자, 뿌리와 녹두 재배토양에서 분리된 Fusarium spp.와 병원성)

  • Paik Su Bong;Do Eun Su
    • Korean Journal Plant Pathology
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    • v.3 no.1
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    • pp.8-12
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    • 1987
  • Fusarium oxysporum, F. moniliforme, F. solani, F. equiseti, F. semitectum, and F. sporotrichioides were detected from seeds, roots and cultivated soil of Phaseolous vidissimus collected from Kyung-gi Provincial Rural Development Administration. The rate of seedling desease incidence was $60\%$ by testing of seed germination using a large petri-dish. According to the blotter method, F. moniliforme showed $7\%$ infection at seed-coat and $2\%$ at cotyledon and embryo. Their pathogenicities of F. moniliforme, F. semitectum, F. equiseti, and F. sporotrichioides isolated from seeds were recognized on seedlings by water-agar test tube methods. F. oxysporum and F. solani isolated from infected-roots had their pathogenicity by water-agar test tube method but were weakly pathogenic by soil treatment method. Their pathogenicities of F. oxysporum. F. solani and F. uiseti isolated from cultivated-soil were recognized by water-agar test tube method. These F. oxysporum and equiseti isolates had their pathogenicities but F. solani was weakly pathogenic by soil treatment method.

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Disinfection of Fusarium-infected Rice Seeds by Prochloraz and Gaseous Chlorine Dioxide

  • Jeon, Young-ah;Lee, Young-yi;Lee, Ho-sun;Sung, Jung-sook;Lee, Seokyoung
    • 한국균학회소식:학술대회논문집
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    • 2014.10a
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    • pp.25-25
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    • 2014
  • Three species of Fusarium, F. fujikuroi, F. verticillioides and F. proliferatum, are known to be associated with bakanae disease of rice [1, 2]. F. fujikuroi infects rice flowers and survive in endosperm and embryo of the seeds. Infected seed is an important source of primary inoculum of pathogens [3]. Seeds of rice (Oryza sativa cv. Boramchan) collected from bakanae-infected field were found to be 96% infected with Fusarium sp., 52% with F. fujikuroi, 42% with F. verticillioides, and 12% with F. proliferatum as determined by incubation method and species-specific PCR assays. F. fujikuroi was detected at lemma/palea, endosperm and embryo whereas F. verticillioides and F. proliferatum were recovered only from lemma/palea by means of component plating test. Seed disinfection methods have been developed to control bakanae disease and prochloraz has been most widely used for rice seeds. Two chemicals formulated with prochloraz (PC 1) and prochloraz + hexaconazole (PC 2) that inhibit biosynthesis of ergosterol strongly reduced the incidence of Fusarium spp. on selective media to 4.7% and 2.0%, respectively. Disease symptoms of rice seedlings in nursery soil were alleviated by chemical treatment; seedlings with elongated leaves or wide angle between leaf and stem were strikingly reduced from 15.6 to 3.2% (PC 1) and 0 (PC 2), stem rots were reduced from 56.9 to 26.2% (PC 1) and 32.1% (PC 2), and normal seedling increased from 0.4 to 13.3% (PC 2). Prochloraz has some disadvantages and risks such as the occurrence of tolerant pathogens [4] and effects on the sterol synthesis in animals and humans [5]. For these reasons, it is necessary to develop new disinfection method that do not induce fungal tolerance and are safe to humans and animals. Chlorine dioxide ($ClO_2$), that is less toxic, produces no harmful byproducts, and has high oxidizing power, has been reported to be effective at disinfection of several phytopathogenic fungi including Colletotrichum spp. and Alternaria spp. [6]. Gaseous $ClO_2$ applied to rice seeds at a concentration of 20 ppm strongly suppressed mycelial growth of Fusarium fujikuroi, F. verticillioides and F. proliferatum. The incidence of Fusarium spp. in dry seed with 8.7% seed moisture content (SMC) tended to decrease as the concentration of $ClO_2$ increased from 20 to 40 ppm. Applying 40 ppm $ClO_2$ at 90% relative humidity, incidence was reduced to 5.3% and resulted in significant reduction of disease symptoms on MS media. In nursery soil, stem rot was reduced from 56.9 to 15.4% and the number of normal seedlings increased from 0.4 to 25.5%. With water-soaked seeds (33.1% SMC) holding moisture in the endosperm and embryo, the effectiveness of disinfection using $ClO_2$ increased, even when treated with only 20 ppm for four hours. This suggests that moisture was a key element for action of $ClO_2$. Removal of the palea and lemma from seeds significantly decreased the incidence of Fusarium spp. to 3.0%. Seed germination appeared to decrease slightly by water-soaking at $30^{\circ}C$ because of increased SMC and by physical damage of embryos from hulling. These results indicate that the use of gaseous $ClO_2$ was effective as a means to disinfect rice seeds infected with Fusarium spp. and that moisture around the pathogens in the seed was an important factor for the action of $ClO_2$. Further investigations should be conducted to ascertain the best conditions for complete disinfection of Fusarium spp. that infect deep site of rice seeds.

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Effects of physico-chemical treatment on 'Nunkeunhukchal' (black sticky rice with giant embryo) for the enhancement of GABA (γ- aminobutyric acid) contents ('눈큰흑찰'의 가바(GABA) 함량 증진을 위한 이화학적 처리 효과)

  • Han, Sang-Ik;Ra, Ji-Eun;Seo, Kyung-Hye;Park, Ji-Young;Seo, Woo Duck;Park, Dong-Soo;Cho, Jun-Hyun;Lee, Jong-Hee;Sim, Eun-Yeong;Nam, Min-Hee
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.59 no.4
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    • pp.398-405
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    • 2014
  • We assessed the GABA accumulation and other components after the 'Nunkeunhukchal (BGE)', 'Josanghukchal (BR)', and 'Ilmibyeo (IB)' grain was soaked in water for 24, 36, 48, 60, 72 and 96 hr. The results showed a continuous accumulation of GABA in soaking treated brown rice of BGE and IB. Among the treated hours, 72 hours of soaking had the maximal accumulation of GABA (51.4 mg/100 g), amino acid, polyphenol and other components. The activities of glutamate decarboxylase (GAD) in brown rice and rice-bran were the same in BGE rice. However, the formation of GABA treated with L-glutamate as substrate showed dramatic increase of 354.6 (fourteen times higher than normal extraction) and 726.4 mg/100 g in BGE rice and rice-bran, respectively. These results suggested that the soaking and extraction with L-glutamate buffer could be better methods for the harvest of increased GABA.

Studies on Proper Medium for Somatic Embryogenesis in Suspension Culture of Rehmania glutinosa and Encapsulation of Somatic Embryos (지황의 현탁배양에서 체세포배 형성에 관여하는 요인분석과 체세포배의 Encapsulation)

  • Park, Ju-Hyun;Park, Sang-Un;Chae, Young-Am
    • Korean Journal of Medicinal Crop Science
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    • v.3 no.2
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    • pp.100-106
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    • 1995
  • This study was conducted to find the factors affecting somatic embryogenesis in suspension culture of Rehmania glutinosa and investigate the possibility of artificial seed production by encapsulation of somatic embryos. Linsmeier-Skoog medium was appeared as proper for somatic embryogenesis. Sucrose with $3{\sim}5%$ as carbon sources was good for somatic embryogenesis, and both ammonium and nitrate nitrogen were necesary for normal somatic embryo production. BA with NAA or kinetin with NAA were better than the use of cytokinin alone for both somatic embryogenesis and numbers of somatic embryos. $AgNO_3$ as protectant for vitrification of seedlings in vitro culture had no harmful effect on somatic embryos. Sphericity of encapsulated seeds was good at 3% gel of sodium alginate but germination was better at 2.5% sodium alginate level. Artificial seeds were germinated and developed normal shoots and roots under in vitro condition.

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Plant Regeneration via in Vitro Culture of Ovule Obtain by Intergeneric Crossing Between Citrus junos Sieb. et Tanaka and Poncirus trifoliata Raf. (유자와 탱자의 속간교잡후 배주배양에 의한 식물체 유기)

  • 이만상;남궁승박
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.6
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    • pp.317-322
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    • 1995
  • As a basic research for breeding new varieties, reciprocal -intergeneric crosses between Citrus junos and P.trifoliata were made. F$_1$ hybrid production using in vitro ovule culture, gametogenesis, and fertilization phenomena were investigated. Frequency of fruit set resulting from crossing of Citrus junos and Poncirus Trifoliata was 16.6% while that of Poncirus Trifoliata and Citrus junos was 11.7%. Callus formation occurred well when ovules at the 6th week after pollination were cultured on MT (Murashige and Tucker) medium supplemented with zeatin 0.5 mg/L and NAA 1.0 or 3.0 mg/L. Immature ovules developed into mature embryos of the MT medium supplemented with 2,4-D 0.1 or 3.0 mg/L. Immature ovules developed into mature embryos of the MT medium supplemented with 2,4 D 0.1 or 0.5 mg/L. The invitro germination rates of 20-week-old ovules set C. junos $\times$ P. Trifoliata and P. Trifoliata $\times$ C. junos were 54.5% and 48.6%, respectively. The emergence ratios of trifoliate hybrids obtained by C. junos $\times$ P. Trifoliata and P. Trifoliata $\times$ C. junos were 56.7% and 100%, respectively. The chromosome number of C. junos and P. Trifoliata was n = 9 or 2n = 18, and the sizes of their pollen grain were 33.75 $\mu$ and 25.0 $\mu$. The length and width of embryo sac in C. junos and P. Trifoliata were 69.38~79.23 $\mu$ and 27.50~38.56 $\mu$, and those of egg cells were 17.50~41.50 $\mu$ and 6.25~8.12$\mu$. Fertilization of C. junos and P. trifoliata terminated 72 h after pollination.

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Acquirement of transgenic rose plants from embryogenic calluses via Agrobacterium tumefaciens (배발생 캘러스를 이용한 아그로박테리움 매개형질전환 장미 식물체 획득)

  • Lee, Su-Young;Lee, Jung-Lim;Kim, Won-Hee;Kim, Seung-Tae;Lee, Eun-Kyung
    • Journal of Plant Biotechnology
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    • v.37 no.4
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    • pp.511-516
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    • 2010
  • The process to acquire intron-GUS gene-expressed transformants from somatic embryos (including embryogenic calli) of Rosa hybrida cv. 'Sweet Yellow' using Agrobacterium-meditated transformation method was reported in this study. Somatic embryos including embryogenic calluses were infected with Agrobacterium tumefaciens AGL1 strain (O.D = 0.7~1.6) including intron-GUS gene for 30 min, and were co-cultured for 3 days. After co-cultivation, they were cultured on embryo germination medium (EGM) supplemented with $250\;mg{\cdot}L^{-1}$ cefotaxim at $4^{\circ}C$ for 7 days. Then, transient GUS gene expression was observed. Shoots were regenerated from the shoot primodia induced from the intron-GUS gene-transferred either somatic embryos or embryogenic calli cultured on EGM supplemented with both cefotaxim $250\;mg{\cdot}L^{-1}$ and ppt $2\;mg{\cdot}L^{-1}$. Before induction of rooting from shoots cultured on shoot growing medium supplemented with both cefotaxim $250\;mg{\cdot}L^{-1}$ and ppt $2\;mg{\cdot}L^{-1}$, the shoots were cultured on multi-shoot induction medium supplemented with both cefotaxim $250\;mg{\cdot}L^{-1}$ and ppt $2\;mg{\cdot}L^{-1}$ to induce multi-shoots. When expression of the gene from a part of the multi-shoots was identified by GUS transient assay, the putative transgenic multishoots were transferred to rooting medium supplemented with cefotaxim $250\;mg{\cdot}L^{-1}$. After the formation of healthy roots, transgenic plantlets were transferred to the greenhouse after acclimatization. The expression rate of the intron-GUS gene in the multi-shoots was 100%.

Water Uptake, Cotyledon Damage after Imbibition and Hypocotyl Elongation in Soybean with Different Seed Size and Color (콩 종실크기 및 종피색에 따른 침종후 수분흡수특성, 자엽손상 및 배축 신장력의 차이)

  • Park, Keum-Yong;Kim, Seok-Dong;Ryu, Yong-Hwan
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.39 no.4
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    • pp.331-338
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    • 1994
  • The experiment was conducted to determine if seed size and seed coat color of soybean might be effective in water uptake and cotyledon damage after imbibition, and hypocotyl elongation. Eight soybean cultivars were separated into two classes of large and small seed based on seed weight, and each class included two cultivars with yellow and black seed color, respectively. Small seed size group was superior in water uptake by seed for 24 hour in imbibition at $25^{\circ}C$ , but its differences decreased as soaking time increased. Small seed cultivars germinated faster and had better germination rate than large ones. However, cultivars with black seed coat showed more slow water uptake at initial time and faster germination than yellow seed, but in 24 hour after imbibition, cultivars with black seed coat had higher water uptake rate than yellow seeds. Small seed cultivar group showed no cotyledon damage in imbibition for 24 hour while large seed cultivars were damaged 78% of cotyledon, and black seed showed low cotyledon damage compared to yellow seed. Hypocotyl length was shorter in large seed rather than in small seed, but hypocotyl thickness in large seed was more thick than in small seed. In correlation coefficients, seed coat rate, embryo rate exhibited significantly negative association with seed weight, and the correlation of seed weight with water uptake in 3 hour after soaking was significantly negative, but in 24 hour showed positive correlation.

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Effects of Temperature on the Oviposition, Feeding and Emergence of the Azuki Bean Weevil (Callosobruchus chinensis L.) in the stored beans (저장두류(貯藏豆類)에 대(對)한 팥바구미의 산란(産卵).섭식(攝食) 및 우화(羽化)에 미치는 온도(溫度)의 영향(影響))

  • Kim, Kyu-Chin;Choi, Hyeon-Soon
    • Korean journal of applied entomology
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    • v.26 no.2 s.71
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    • pp.71-81
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    • 1987
  • This study was conducted to study the effects of temperature on the oviposition, feeding and emergence of the azuki bean weevil(ABW) (Callesobruchus chinensis L.) in the stored beans. And ovipositional preference, hatching and emergence on the preference and non-preference hosts were also investigated. ABW has four generations a year and overwinters as larva stage in the bean grains. Optimum oviposition temperature of the ABW ranges from $25^{\circ}C\;to\;30^{\circ}C$. Oviposition preference in different hosts was in the order of azuki bean, mung bean, soy bean, kidney bean, black-soybean, pea. At $25^{\circ}C$, optimum development temperature emergence of period the ABW averaged 29 days on azuki bean, 31 days on mung bean, 49 days on soy bean, 46 days on black-soybean, 34 days on Pea. Percent hatch in different hosts was in the order of azuki bean, mung bean, soy bean, pea, black-soybean, kidney bean. But percent emergence was in the order of azuki bean, mung bean, pea, black-soybean, kidney bean. Especially, in azuki bean and mung bean, percentage of hatching and emergence was very high. And in soy bean and black-soybean, percentage of hatching was high but emergence was low. The kidney bean was hatching but not emergence at all. The longevity at the different temperatures appeared to be $4{\sim}5$ days at $35^{\circ}C,\;5{\sim}7$ days at $25^{\circ}C{\sim}30^{\circ}C$, $10{\sim}15$ days at $20^{\circ}C\;and\;19{\sim}24$ days at $15^{\circ}C$. Kidney bean of feeding non-preference host oviposited at Seed coat and Endosperm, but no larva developed. And cotyledon and embryo parts were not oviposited at all. The percentage of germination of egg laid bean grain and damaged bean grains was low. Especially, the percentage of germination of damaged bean grains was 0% on azuki bean, 5% on black-soy bean, 7% on Pea.

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