• Title/Summary/Keyword: rice callus

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Improvement of cultural efficiency using DNA markers in anther and seed culture of rice (DNA marker를 이용한 벼의 조직배양 효율개선)

  • Kim, Hong-Jib;Kim, Tae-Heon;Sohn, Jae-Keun
    • Current Research on Agriculture and Life Sciences
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    • v.27
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    • pp.21-28
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    • 2009
  • The purpose of this study was to improve the culturability of 'IR 36', a indica type rice cultivar using DNA marker associated with the ability of plant regeneration in anther and seed culture. The culturability of 6 rice cultivars and 2 indica/japonica lines ('MGRI 036', 'MGRI 079') were investigated in anther and seed culture. The culturability of 3 japonica rice cultivars were much higher than tongil and indica rice cultivars, and 'MGRI 036' and 'MGRI 079' has high culturability with 20% regenerability, also. 34 $BC_2F_4$ 4 lines were selected by marker screening using RZ400 among 90 $BC_2F_4$ lines derived from a cross between 'MGRI 079' and 'IR 36'. The frequency of callus formation of 10 $BC_2F_4$ lines were higher than 'IR 36' in anther culture among selected 34 $BC_2F_4$ lines. The ability of plant regeneration of 10 lines were higher than 'IR 36' in the seed culture among selected 34 $BC_2F_4$ lines. A promising line, $BC_2F_4$-28, was selected to have better culturability in the anther and seed culture among selected 34 $BC_2F_4$ lines. The heading date and grain shape of the $BC_2F_4$-28 was similar to 'IR 36'. Using the RZ400 DNA marker associated with the culturability will be useful method for improving of indica rice culticvar's culturability in rice breeding program.

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Enhanced proline accumulation and salt stress tolerance of transgenic indica rice by over-expressing P5CSF129A gene

  • Kumar, Vinay;Shriram, Varsha;Kishor, P.B. Kavi;Jawali, Narendra;Shitole, M.G.
    • Plant Biotechnology Reports
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    • v.4 no.1
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    • pp.37-48
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    • 2010
  • [ ${\Delta}^1$ ]pyrroline-5-carboxylate synthetase (P5CS) is a proline biosynthetic pathway enzyme and is known for conferring enhanced salt and drought stress in transgenics carrying this gene in a variety of plant species; however, the wild-type P5CS is subjected to feedback control. Therefore, in the present study, we used a mutagenized version of this osmoregulatory gene-P5CSF129A, which is not subjected to feedback control, for producing transgenic indica rice plants of cultivar Karjat-3 via Agrobacterium tumefaciens. We have used two types of explants for this purpose, namely mature embryo-derived callus and shoot apices. Various parameters for transformation were optimized including antibiotic concentration for selection, duration of cocultivation, addition of phenolic compound, and bacterial culture density. The resultant primary transgenic plants showed more enhanced proline accumulation than their non-transformed counterparts. This proline level was particularly enhanced in the transgenic plants of next generation ($T_1$) under 150 mM NaCl stress. The higher proline level shown by transgenic plants was associated with better biomass production and growth performance under salt stress and lower extent of lipid peroxidation, indicating that overproduction of proline may have a role in counteracting the negative effect of salt stress and higher maintenance of cellular integrity and basic physiological processes under stress.

Variations of Agronomic Traits on the Progenies of the Different Callus Origin and Subculture Period in Rice (벼에서 캘러스 유기원와 계대배양 기간에 따른 재분화 계통의 주요 농업형질 변이)

  • Yi, Gi-Hwan;Oh, Byeong-Geun;Yang, Sae-Jun;Kim, Soon-Chul;Nam, Min-Hee
    • Journal of Plant Biotechnology
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    • v.30 no.4
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    • pp.365-368
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    • 2003
  • Variations of agronomic traits were evaluated on the progenies of regenerated rice plants of different callus origin and subculture period. From eighty-eight percent to ninety percents of ovary culture-derived plants (OCP) and anther culture-derived plants (ACP) were not segregated within the lines in major agronomic characters. Compare to ACP and OCP lines, the seed culture derived lines (SCP) showed more segregation (17%) in major agronomic traits among the lines. The most frequent segregating traits were grain fertility (6.1%) in ACP, leaf color (4.2%) in OCP and grain fertility (4.9%) and leaf color (4.9%) in SCP lines. The SCP line showed more variation in culm length, panicle length and heading date than those of OCP and ACP lines. The variation of agronomic traits in SCP lines was tended to increase with prolonged subculture. Culm and panicle length were shorter than those of original cultivar in all three types of tissue culture-derived lines (OCP, ACP and SCP).

Effects of Mixing Ratio of Substrates and Concentration of Fertigation in Nutrient Solutions on the Early Growth of Cucumber Plug Seedlings (배지의 혼입비율과 관비 양액 농도가 오이 플러그묘의 초기생장에 미치는 영향)

  • Kim, Hong-Gi;Cho, Ja-Yong;Bae, Jong-Hyang;Yang, Seung-Yul;Heo, Buk-Gu
    • Journal of Bio-Environment Control
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    • v.16 no.2
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    • pp.101-107
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    • 2007
  • This experiment was carried out to develop the substrate mixtures for the production of high quality plug seedlings of cucumber. Peatmoss based substrates, rice hull, carbonized rice hull, decomposed sawdust, perlite and granular rockwool were mixed by five different mixing ratioes (M1, M2, M3, M4 and M5). The cultivars used were cucumber (Cucumis sativus L. cv. Janghyung heukjinju) plants. The higher the content of peatmoss added, the higher the plant growth in terms of plant height, leaf area and total dry weight, which leading to the production of high quality plug seedlings. Seedlings growth of cucumber were greater in M5 mixtures [peatmoss:rice hull:decomposed sawdust=40:40:20(v/v)], M4 mixtures [peatmoss:rice hull:decomposed sawdust:granular rockwool=30:25:20:25(v/v)] and M2 mixtures [peatmoss:rice hull:decomposed sawdust:granular rockwool=20:20:15:25:20(v/v)] The concentrations of nutrient solution (EC) had a great influence on plant height, leaf area, total fresh and dry weight of cucumber seedlings growth. As the concentration of nutrient solution increased from 0.1 to $1.5dS{\cdot}m^{-1}$, the growth and seedling quality of cucumber in plant height, leaf area and dry weight were significantly improved.

Effects of the Mixing Ratio of the Different Substrates and the Concentration of Fertigation in Nutrient Solution on the Growth of Tomato Plug Seedlings (배지의 혼합비율과 관비 양액 농도가 토마토 플러그묘의 생장에 미치는 영향)

  • Kim, Hong-Gi;Cho, Ja-Yong;Yu, Sung-Oh;Yang, Seung-Yul;Kang, Jong-Gu;Heo, Buk-Gu
    • Journal of Bio-Environment Control
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    • v.16 no.2
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    • pp.108-114
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    • 2007
  • This study was conducted to clarify the effects of the different mixing ratios of substrate mixtures based on peat moss and the concentration of nutrient solution on the growth of tomato (Lycopersicon esculentum Mill.) seedlings. Substrates such as peat moss, rice hull, carbonized rice hull, decomposed sawdust, perlite and granular rock wool were mixed and used. The concentration of nutrient solution were adjusted to EC $0.5{\sim}1.5mS/cm$. The volumetric moisture contents became higher as peat moss mixed were much more. Total porosities in all substrate mixtures were over 80%, and pH in substrate mixtures became lower as the volume of peat moss mixed higher. Mixing ratios of substrates suitable for the production of tomato seedlings with the higher quality were peat moss:rice hull:carbonized rice hull:decomposed sawdust:perlite=25:10:25:20:20(v/v). The plant growth was not significant among the different substrate mixtures. However, plant growth such as plant height, leaf area, and total dry weight became significantly increased as EC increasing.

Characterization of the 5-methyltryptophan Resistant Mutant Lines Selected by Mutagenized Seeds in Rice (돌연변이 벼 종자로부터 선발된 5-methyltryptophan 저항성 계통의 특성)

  • 이효연;배창휴;임용표;박노동;조백호;이수인;최해춘;김호일
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.6
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    • pp.453-459
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    • 2000
  • Three rice (Oryza sativa L. var Dong-Jin) mutants (DTR1, DTR2, DTR3) resistant to S-methyltryptophan (5MT) were selected by mutagenized M3 seeds. The frequency of chlorophyll mutations induced by the EMS (0.2%) treatment performed 2 hours after flowering is clearly higher than that induced by other treatments in M1 generation. Progeny obtained from the self-pollinating of 5MT-resistant lines segregated with 3 : 1 of resistant to sensitive ratio. Furthermore, the ratio of homozygote to heterozygote in 5MT-resistant plants of the M4 generation was 1:2. These results show that 5MT resistance was inherited as a single dominant nuclear gene. The resistance was also expressed in callus derived from seeds. Total free amino acid content in homozygous seeds of DTR1 and DTR2 showed about 1.7 fold-increased compared to the wild-type seeds. In particular, the levels of phenylalanine and Iysine were, respectively, 6.2 and 3.2 times higher than those in the wild-type seeds. However, seeds of DTR3 had lower levels of free amino acid than the wild-type seeds. This result indicate that these mutants as a significant step towards the production of new rice with balanced amino acid content.

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Development of Bialaphos-Resistant Transgenic Rice Using Agrobacterium tumefaciens (Agrobacterium tumefaciens를 이용한 bialaphos 저항성 형질전환 벼의 개발)

  • 이효연;이춘환;김호일;한원동;최지은;김진호;임용표
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.4
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    • pp.283-288
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    • 1998
  • The bialaphos is a potent inhibitor of glutamine synthease in higher plants and is used as a non-selective herbicide. We have used the bialaphos resistant gene(Bar) encoding for an acetyltransferase isolated from Streptomyces hygroscopicus SF1293. Callus derived from mature seeds of rice(Oryza sativa L. cv. Dong Jin) were co-cultivated with Agrobacterium tumefaciens EHA101 carring a plasmid pGPTV-HB containing genes for hygromycin resistance (HygR) and Bar. Transgenic plants showing in vitro resistance to 50 mg/L hygromycin and 10 mg/L bialaphos were obtained by using a two-step selection/regeneration procedure. Transformation efficiency of rice was about 30% which was as high as reported in other dicotyledons. Progenies ($\textrm{T}_{1}$ generation) derived from primary transformant of 17 lines were segregated with a 3 resistant : 1 sensitive ratio in medium containing hygromycin and bialaphos. Stable integration of Bar gene into chromosomal DNA was proven by Southern blot analysis of genomic DNA isolated from $\textrm{T}_{2}$ progenies. Transgenic plants ($\textrm{T}_{3}$) grown in the field were resistant to bialaphos (Basta) at a dosage lethal to wild type plants.

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Bioassays of Plant Hormones and Plant Growth Regulating Substances I . Auxins, Gibberellins, and Cytokinins (식물홀몬 및 생장조절물질의 생물검정기술 I. 옥신, 지베렐린 및 싸이토키닌)

  • 이정명
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.34 no.s01
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    • pp.4-15
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    • 1989
  • The objective of this paper is to compare and summarize the procedure and effectiveness of some bioassay systems and to point out ways to obtain reliable results from each bioassay. Detailed C:escriptions were given for those widely-adapted bioassay methods, such as mungbean rooting (auxin), Avena first internode straight growth (auxin), dwarf rice growth (gibberellin), dwarf pea epicotyl elongation (gibberellin), radish cotyledon expansion test (cytokinin), and tobacco stem pith callus growth (cytokinin), and the effects of various plant growth regulators including some recently introduced growth retardants (Paclobutrazol, Uniconazol, etc.) were also summarized.

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Studies on the Viability of Cultured Anther in Rice Anther Culture I. Changes of Respiratory Activity by Genotype and Cold-pretreatment (벼 배양약에서 약의 활력 연구 I. 품종 및 저온 전처리에 따른 호흡활성의 변화)

  • Seung Yeob, Lee;Seon Yong, Lee;Jang Soo, Choi
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.33 no.3
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    • pp.248-253
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    • 1988
  • The longer pollen stage grew to flowering stage, the higher anther respiratory rate in vivo became. and it was rapidly increased just before flowering. The anther respiratory rate in vitro showed the first and second peak points after 3-7 days and 9-1l days in culture, respectively, and fastest and highest in Daecheongbyeo with high sporophytic potentiality. It was lower in cold-pretreatment than non-treatment at the early days, but higher from 15 days after culture. The frequency of browning anthers was promoted by cold-pretreatment. The respiratory rate was not different between uncolored and browned anthers at 12 days, but it was higher in browned anthers after 24 days in culture.

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Characterization of Salt Tolerant Rice Mutant Lines Derived from Azetidine-2-Carboxylic Acid Resistant Cell Lines Induced by Gamma Ray Irradiation (AZCA 저항성 돌연변이 세포주로부터 선발 육성만 내염성 벼 돌연변이 계통의 특성 검정)

  • Song, Jae-Young;Kim, Dong-Sub;Lee, Geung-Joo;Lee, In-Sok;Kang, Kwon-Kyoo;Yun, Song-Joong;Kang, Si-Yong
    • Journal of Plant Biotechnology
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    • v.34 no.1
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    • pp.61-68
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    • 2007
  • To develop rice (Oryza sativa L.) cultivars to be planted on salt-affected sites, cell lines with enhanced proline content and resistance to growth inhibition by Azetidine-2-carboxylic acid (AZCA), a proline analogue, were screened out among calli irradiated with gamma ray of 50, 70, 90, and 120 Gy. The calli had been derived from embryo culture of the cultivar Donganbyeo. Selected AZCA resistant lines that had high proline accumulation were used as sources for selection of NaCl resistant lines. To determine an optimum concentration for selection of NaCl resistant lines, Donganbyeo seeds were initially cultured on the media containing various NaCl concentrations (0 to 2.5%) for 40 days, and 1.5% NaCl concentration was determined as the optimum concentration. One hundred sixteen salt-tolerant (ST) lines were selected from bulked 20,000 seeds of the AZCA resistant $M_{3}$ seeds in the medium containing 1.5% NaCl. The putative 33 lines ($M_{4}$ generation) considered with salt-tolerance were further analyzed for salt tolerance, amino acid and ion contents, and expression patterns of the salt tolerance-related genes. Out of the 33 lines, 7 lines were confirmed to have superior salt tolerance. Based on growth comparison of the entries, the selected mutant lines exhibited greater shoot length with average 1.5 times, root length with 1.3 times, root numbers with 1.1 times, and fresh weight with 1.5 times than control. Proline contents were increased maximum 20%, 100% and 20% in the leaf, seed and callus, respectively, of the selected lines. Compared to control, amino acid contents of the mutants were 24 to 29%, 49 to 143%, 32 to 60% higher in the leaf, seed and callus, respectively. The ratio of $Na^{+}/K^{+}$ for most of the ST-lines were lower than that of control, ranging from 1.0 to 3.8 for the leaf and 11.5 to 28.5 for the root, while the control had 3.5 and 32.9 in the leaf and root, respectively. The transcription patterns for the P5CS and NHXI genes observed by RT-PCR analysis indicated that these genes were actively expressed under salt stress. The selected mutants will be useful for the development of rice cultivar resistant to salt stress.