• Title/Summary/Keyword: rice progenies

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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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Phenotypic and Genotypic Analysis of Rice Lesion Mimic Mutants

  • Matin, Mohammad Nurul;Pandeya, Devendra;Baek, Kwnag-Hyun;Lee, Dong-Sun;Lee, Jai-Heon;Kang, Ho-Duck;Kang, Sang-Gu
    • The Plant Pathology Journal
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    • v.26 no.2
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    • pp.159-169
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    • 2010
  • Lesion mimic mutant (LMM) plants display spontaneous necrotic lesions on their leaves without any pathogenic infection. Specific rice LMMs designated as spotted leaf (spl) including spl1, spl3, spl4, spl5 and spl6 are genetically known as lesion resembling disease (lrd) mutant. The inheritance patterns in the $F_1$ and $F_2$ progenies of these mutants are controlled by recessive genetic factors. Lesion development in the rice LMMs were controlled by both development stages and environmental factors. The rice LMMs exhibited higher numbers of spots under $45^{\circ}C$ temperature than those under $30^{\circ}C$. Contents of chlorophyll were drastically reduced at 60 days old LMM leaves when the spot formation was severe. The levels of endogenous hydrogen peroxide were highest at 45 days old mutants but reduced at 60 days old. Transcription levels of stress related genes including thioredoxin peroxidase and protein disulfide isomerase were reduced in spotted leaves than those of non spotted leaves. It could be suggested that scavenging system against reactive oxygen species induced by either stresses or innate metabolisms may not work properly in the rice LMMs. As these rice LMMs autonomously expressed clear lesions of lrd phenotype without pathogen infection, it could be useful to understand stresses responses in plants.

Salinity Tolerance of Progenies between Korean Cultivars and IRRI's New Plant Type Lines in Rice

  • Lee, Seung-Yeob;Dharmawansa Senadhira
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.43 no.4
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    • pp.234-238
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    • 1998
  • To select new germplasm for salinity tolerance from new plant type (NPT) breeding lines, the sixty F$_4$ lines selected from the crosses between Korean cultivars and IRRI's NPT lines were evaluated for salinity tolerance at the seedling stage with salinized culture solution (EC=12 dS/m) in the controlled conditions. Two NTP lines derived from a cross between 'Ilmibyeo' and 'IR66152-AC5-1', 'HR15258-7-1' and 'HR15258-27-1', were found to have good tolerance. The salinity tolerance of the lines was compared to their parents and the sensitive ('IR29') and tolerant ('Pokkali') checks in three salinity levels, no salinity (control) and an EC of 12 and 16 dS/m. Visual salinity score, shoot Na+ and Na-K ratio in two NPT lines was significantly low compared with the parents and IR29. Indicating that salinity tolerance of the lines might be derived from a transgressive segregation. The relative water content of the lines was higher than Pokkali, and the dry weight of shoot and root was proportionally decreased to salinity score and salinizing concentration. The visual salinity scores were significantly correlated with shoot Na concentration, Na-K ratio, relative water content, and reduction of dry weight (P<0.01). Their tolerance was attributed to root and shoot characteristics that led to high shoot water content, thus diluting the toxic effect of salts.

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Molecular biological characteristics and analysis using the specific markers of leaf folder-resistant GM rice (혹명나방저항성 GM 벼의 분자생물학적 특성 및 특이 마커를 이용한 검정)

  • Shin, Kong-Sik;Lee, Si-Myoung;Lim, Sun-Hyung;Woo, Hee-Jong;Cho, Hyun-Suk;Lee, Kyeong-Ryeol;Lee, Myung-Chul;Kweon, Soon-Jong;Suh, Seok-Cheol
    • Journal of Plant Biotechnology
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    • v.36 no.2
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    • pp.115-123
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    • 2009
  • In recent years, several genetically modified (GM) crops have been developed worldwide through the recombinant DNA technology and commercialized by various agricultural biotechnological companies. Commercialization of GM crops will be required the assesment of risks associated with the release of GM crops. In advance of the commercial release of GM crops, developer should submit the several information on GM crops for approval. In this study, we carried out to provide the molecular data for the risk assessment of GM rice containing insect-resistant gene, modified Cry1Ac (CryIAc1). Through the molecular analysis with CryIAc1 induced GM rice, we confirmed the steady integration and expression of transgene, the transgene copy number, the adjacent region sequences of inserted gene into rice genome, and the transgene stability in progenies. For the qualitative PCR detection methods, specific primer pairs were designed on the basis of integration sequences, and construct- and event-specific detection markers were developed for leaf folder-resistant rice, Cr7-1 line. From these results, we demonstrated that the molecular data and the PCR detection methods of leaf folderresistant GM rice could be acceptable to conduct the biosafety and environment risk assessment.

Agronomic characteristics of stay-green mutant derived from an early-maturing rice variety 'Pyeongwon'

  • Won, Yong-Jae;Ji, Hyeon-So;Ahn, Eok-Keun;Lee, Jeong-Heui;Jung, Kuk-Hyun;Lee, Sang-Bok;Hong, Ha-Cheol;Hyun, Ung-Jo;Ha, Woon-Goo;Kim, Myeong-Ki;Kim, Byeong-Ju
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.72-72
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    • 2017
  • We found a new stay-green mutant from 'Pyeongwon' which is an early-maturing rice variety in Korea. The mutant showed green leaves after grain ripening period and it maintained higher SPAD value than wild type rice plant and original variety 'Pyeongwon'. The stay-green trait in rice, three genes have been identified up to date. The non-yellow coloring1 (NYC1) gene encodes a chloroplast-localized short-chain dehydrogenase/reductase (SDR) with three transmembrane domains. The non-yellow coloring3 (NYC3) gene encodes a plastid-localizing alpha/beta hydrolase-fold family protein with an esterase/lipase motif. The Sgr gene encodes a novel chloroplast protein and regulates the destabilization of the light-harvesting chlorophyll binding protein (LHCP) complexes of the thylakoid membranes, which is a prerequisite event for the degradation of chlorophylls and LHCPs during senescence. After sequencing the PCR products, we found a single nucleotide variation($A{\rightarrow}T$) in the NYC1 gene, which changes the amino acid lysine to methionine. The NYC1 gene encodes a short-chain dehydrogenase/reductase(SDR) protein. And we confirmed the co-segregation between SNP and stay-green trait from genotyping the progenies of the mutant.

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Mapping quantitative trait loci controlling low-temperature germinability in rice

  • Nguyen, Hoang Nam;Park, In-Kyu;Yeo, Sang-Min;Yun, Yeo-Tae;Ahn, Sang-Nag
    • Korean Journal of Agricultural Science
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    • v.39 no.4
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    • pp.477-482
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    • 2012
  • Low-temperature germination is one of the major determinants for stable stand establishment in the rice direct seeding method in temperate regions and at high altitude areas. Quantitative trait loci (QTL) controlling low-temperature germinability in rice were identified using 96 introgression lines (ILs) derived from a cross between Oryza rufipogon and the Korean japonica cultivar, 'Hwaseongbyeo'. The germination rate at $15^{\circ}C$ was measured to represent low-temperature germination and used for QTL analysis. The germination rate at $15^{\circ}C$ for 7 days of Oryza rufipogon and Hwaseongbyeo was 93.3 and 28.7%, respectively, and that of progenies ranged from 0 to 48%. A linkage map was constructed using 135 simple sequence repeat (SSR) markers. Five putative QTLs associated with low-temperature germination were detected on chromosomes 1, 3, 4, 10 and 11. The QTL, qltg10 on chromosome 10 accounted for 19.2% of the total phenotypic variation for low-temperature germinability. Four additional QTL, accounted for 10.4 - 15.1% of the total phenotypic variation. The O. rufipogon alleles in all detected QTLs loci increased the low-temperature germination rate. No QTL associated with low temperature germinability has been detected near the qltg10 QTL in this study suggesting that qltg10 is a new QTL. The locus, qltg10 is of particular interest because of its independence from undesirable height and maturity effects. The DNA markers linked to the QTL for low temperature germinability would be useful in selecting lines with enhanced low temperature germinability in rice breeding program.

Generation and DNA Characterization of High-lysine Mutants by Biochemical Selection from Callus Culture of 'Hwayeongbyeo'

  • Yi Gi-Hwan;Choi Jun-Ho;Kim Kyung-Min;Jeong Eung-Gi;Park Hyang-Mi;Kim Doh-Hoon;Ku Yeon Chung;Eun Moo-Young;Kim Ho-Yeong;Nam Min-Hee
    • Plant Resources
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    • v.8 no.1
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    • pp.60-66
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    • 2005
  • Lysine is the first essential amino acid for optimal nutrient quality in rice grain. For the narrow genetic diversities of lysine contents in rice, somaclonal variation was the source of mutation in our breeding program. Biochemical selection was conducted using 1 mM S-(2-aminoethyl) cysteine followed by two passages of 5 mM lysine plus threonine in the callus subculture medium. The lysine contents in endosperm of all progenies recovered from the biochemical selection were higher than those of their donor cultivar 'Hwayeongbyeo'. These elevated lysine levels of mutants were successfully transmitted to $M_4$ generation. The lysine contents in endosperm varied 3.85 to $4.80\%$ compare to their donor cultivar 'Hwayeongbyeo' was $3.85\%$. Three of high-lysine germplasms, Lys-l, Lys-2 and Lys-7 were selected by biochemical selection and rapid screening methods. DNA analysis showed that a new insertion of Tos 17 which mapped to rice chromosome 11 on the high-lysine mutant, Lys-2.

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Genetic Insights into Domestication Loci Associated with Awn Development in Rice

  • Ngoc Ha Luong;Sangshetty G. Balkunde;Kyu-Chan Shim;Cheryl Adeva;Hyun-Sook Lee;Hyun-Jung Kim;Sang-Nag Ahn
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.33-33
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    • 2022
  • Rice (Oryza sativa L.) is a widely studied domesticated model plant. Seed awning is an unfavorable trait during rice harvesting and processing. Hence, awn was one of the target characters selected during domestication. However, the genetic mechanisms underlying awn development in rice are not well understood. In this study, we analyzed the genes for awn development using a mapping population derived from a cross between the Korean indica cultivar 'Milyang23' and NIL4/9 (derived from a cross between 'Hwaseong' and O. minuta). Two quantitative trait loci (QTLs), qAwn4 and qAwn9 were mapped on chromosome 4 and 9, respectively, increased awn length in an additive manner. Through comparative sequencing analyses parental lines, LABA1 was determined as the causal gene underlying qAwn4. qAwn9 was mapped to a 199-kb physical region between markers RM24663 and RM24679. Within this interval, 27 annotated genes were identified, and five genes, including a basic leucine zipper transcription factor 76 (OsbZIP76), were considered candidate genes for qAwn9 based on their functional annotations and sequence variations. Haplotype analysis using the candidate genes revealed tropical japonica specific sequence variants in the qAwn9 region, which partly explains the non-detection of qAwn9 in previous studies that used progenies from interspecific crosses. This provides further evidence that OsbZIP76 is possibly a causal gene for qAwn9. The O. minuta qAwn9 allele was identified as a major QTL associated with awn development in rice, providing an important molecular target for basic genetic research and domestication studies. Our results lay the foundation for further cloning of the awn gene underlying qAwn9.

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Evaluation of Pollen Viability of Nakdongbyeo, Two Transgenic Rice Lines, Its Hybrids with Weedy Rice, and Subsequent Selfed Progenies: F2 and F3 (낙동벼, 2개의 promoter를 각각 삽입한 유전자변형 계통과 잡초성벼(Oryza sativa)인공수정 한 후 다음세대인 F1, F2, F3의 화분활력 평가)

  • Ghimire, Sita Ram;Sohn, Eun-Young;Shin, Dong-Hyun;Lee, In-Jung;Kim, Kil-Ung
    • Journal of Life Science
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    • v.19 no.7
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    • pp.839-844
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    • 2009
  • This experiment was conducted to evaluate pollen viability of Nakdongbyeo, transgenic rice lines, an F$_1$ hybrid from a cross between Milyang weedy rice and ABC-promoter transgenic rice line containing basta-resistant (bar) gene and subsequent selfed progenies, F$_2$ and F$_3$. The reaction of pollen with 3-{4,5 dimethylthiazolyl-2}-2,5-diphenyl monotetrazolium bromide (MTT) as a staining chemical immediately after pollen shedding showed maximum pollen viability of 86% in Nakdongbeyo, 75% in ABC-promoter transgenic rice line, 62% in ubiquitin-promoter transgenic line, 68% in F$_1$, 79% in F$_2$ and 78% in F$_3$. Viability gradually declined during subsequent observations at 20-minute intervals. However, there was a drastic decline in pollen viability after 40 minutes of pollen shedding. The mean difference of pollen viability among rice lines and time was highly significant, indicating significantly different pollen viabilities at different time intervals. Maximum viability of 36.2% was observed in F$_3$ and minimum viability of 3.5% was found in F$_2$ at 90 min after pollen shedding. Results of this experiment on pollen viability and longevity elucidate potential risks of pollen-mediated flow of herbicide-resistant gene from transgenic rice lines and possible integration of it into the weedy rice population.

Genetic Analysis on the Bacterial Blight Resistance of Suweon497, a Rice Breeding Line Developed through Wide Hybridization (벼 종간교잡 후대계통 '수원497호'의 흰잎마름병 저항성에 대한 유전분석)

  • Jeung, Ji Ung;Roh, Tae Hwan;Kang, Kyung Ho;Jeong, Jong Min;Kim, Myeong Ki;Kim, Yeon Gyu
    • Korean Journal of Breeding Science
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    • v.43 no.1
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    • pp.81-91
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    • 2011
  • Rice wild relatives have been recognized as reservoirs of genetic reinforcements to improve cultivating rice against biotic and abiotic stresses. A wild relative, Oryza. minuta(BBCC; Acc. 101141), was hybridized with a Korean Japonica cultivar, 'Hwaseong'(AA), followed by ovule culture and several times of back crossings to overcome high level of sterility. During evaluation of the introgression lines, breeding line exhibited resistance to bacterial blight with reasonable agronomic performances, and nominated as an elite breeding line, the 'Suweon497'. A mapping population, to dissect genetic basis of the resistance, was constructed by using $F_2$ progenies of the 'Suweon497' ${\times}$ 'Milyang23'. Association analysis between SSR marker genotypes and pathogenisity levels of each $F_2$ progeny revealed the end terminal region of rice chromosome 11 as the nesting place for the wild rice derived bacterial blight resistance gene, where at least four other genes, Xa3, Xa4, Xa26 and Xa31, have been reported.