• Journal of Plant Biotechnology
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• 제5권1호
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• pp.51-57
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• 2003

Doubled haploids have long been recognized as a valuable tool in plant breeding since it not only offers the quickest method of advancing heterozygous breeding lines to homozygosity, but also increased the selection efficiency over conventional procedures due to better discrimination between genotypes within any one generation. Salt tolerant mutants were obtained in rice the variety, 'Hawsungbyeo', through in vitro mutagenesis of in vitro cultured anther-derived calli. Various doses (30, 50, 70 and 90 Gy) of gamma ray were applied to investigate the effect of radiation on callus formation on medium containing 1% NaCl, green plant regeneration, frequency of selected doubled haploid mutants and of the salt tolerant screen. It was demonstrated that the dose of 30 and 50 Gy gamma rays had significant effects on callus formation, regeneration and selection of salt tolerance. No tolerant lines were obtained from non-mutagenized cultures. From gamma ray irradiated cultures, five tolerant lines ($M_2$generation) at germination stage and 13 tolerant lines ($M_3$genoration) at seedling stage were obtained. The frequency of salt tolerant mutants indicates that anther culture applied in connection with gamma rays is an effective way to improve salt tolerance.

• Molecules and Cells
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• 제26권4호
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• pp.368-372
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• 2008

Recent molecular and genetic studies in rice, a short-day plant, have elucidated both conservation and divergence of photoperiod pathway genes and their regulators. However, the biological roles of rice genes that act within the autonomous pathway are still largely unknown. In order to better understand the function of the autonomous pathway genes in rice, we conducted molecular genetic analyses of OsFVE, a rice gene homologous to Arabidopsis FVE. OsFVE was found to be ubiquitously expressed in vegetative and reproductive organs. Overexpression of OsFVE could rescue the flowering time phenotype of the Arabidopsis fve mutants by up-regulating expression of the SUPPRESSOR OF OVEREXPRESSION OF CO1 (SOC1) and down-regulating FLOWERING LOCUS C (FLC) expression. These results suggest that there may be a conserved function between OsFVE and FVE in the control of flowering time. However, OsFVE overexpression in the fve mutants did not rescue the flowering time phenotype in in relation to the response to intermittent cold treatment.

• Animal cells and systems
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• 제2권1호
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• pp.81-86
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• 1998

Recent identification and characterization of plant homeobox genes suggest that they play important roles in morphogenetic events. OSH1, one of the rice homeobox genes, is thought to be related to organ development since the changes of OSH1 gene expression cause morphological abnormalities of leaves by the ectopic expression and is expressed during early embryogenesis. In this experiment, the expression pattern of OSH1 was analyzedinmutants by in situ hybridization, and OSH1's potential as a molecular marker was explored. Region-specific expression of OSH1 during early embryogenesis shows that OSH1 could be used as a molecular marker for characterizing embryo mutants. Although several organless and shootless mutants showed normal expression of OSM1, some mutants exhibited abnormal expression patterns. In a minute organless cle1-1 embryo whose epidermis resembled morphologically the epithelium of scutellum, OSH1 expression was limited to a small basal region. This expression pattern suggests the gross deletion of the basal part. In a radicleless mutant, odm115, OSH1 expression was detected in a basal region instead of subcentral region of the ventral side. Together with other characteristics (short embryo and normal adventitious roots), odm115 was estimated to be derived from the deletion of basal region. Among five shootless mutants, three showed normal expression of OSH1. In the shl2 embryo, no expression of OSH1 was observed. In the shl1 embryo, however, OSH1 expression was extended to a dorsal side, indicating that SHL2 might be related to dorsoventral patterning. The above results of in situ hybrydization clearly indicate that OSH1 can be utilized as a marker for characterizing gene functions of embryo mutants.

• 한국작물학회지
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• 제38권3호
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• pp.264-274
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• 1993

본 연구는 화청벼와 IR24의 돌연변이 계통인 분상질 및 심백 변이체의 작물학적 및 이화학적특성과 제병 및 알콜발효특성을 조사하여 그 가공적 이용성을 검토하고 분상질배유 유전자의 유전분리와 표지인자들과의 연관 관계를 밝히고자 수행하였으며 그 결과는 다음과 같다. 1. 돌연변이체의 종실은 원품종에 비해 작았으며, 천입중과 1$\ell$중도 전반적으로 가벼운 것으로 나타났다. 2. 변이체들의 출수기는 원품종과 같았으며, 정조수량은 원품종에 비해 낮았다. 3. 분상질 배유의 전분립은 느슨한 결정구조를 갖고 있었고, 전분립의 크기는 원품종과 변이체들간에 동일경향이 아니었으며, 원품종에 비해 변이체들은 밀도와 경도가 낮았다. 4. 변이체들의 아밀로스 함량은 16.9～28.5%로 변이복이 크게 나타났다. 현미의 조단백 함량에서 화청벼의 분상질변이체들은 원품종과 같았으나 IR24의 심백변이체인 47106은 원품종(8.30%)에 비해 11.32%로 높게 나타났다. 5. Gel consistency는 원품종들이 soft의 특성을 보였으나 분질 및 심백의 변이체에서는 모도 medium의 특성을 보였고, 백미의 알칼리 붕괴도는 원품종에 비해 분수질 변이체들에서 높게 나타났다. 6. Amylogram에 의한 호화개시 온도는 IR24의 분상질변이체(47160)를 제외하고 모도 원품종과 비슷하였으나, 최고점도와 최저점도 및 굳음성은 원품종 화청벼와 IR24에 비해 변이체에서 월등히 낮게 나타났다. 7. Instron으로 측정된 증편의 경도와 gumminess로 볼 때 화청벼 분상질 및 심백변이체로 조제된 증편은 다소 부드러운 특성을 나타냈으나, IR24의 경우는 일정 경향이 없었다. 분상질미와 심백미의 증편부피는 원품종에 비해 대체로 크게 나타났다. 8. 알콜발효 과정 중 당도 저하는 변이체들 특히 분상질에서 빨리 일어났고, 발효 10일 후의 알콜생성량도 원품종에 비해 변이체들에서 높게 나타나서 분상질미는 양조용으로서의 이용성이 크게 기대되었다. 9. 화청벼와 IR24에서 유래한 분상질 배유유전자들의 대립성 검정결과 서로 다른 3개의 유전자좌를 탐색하였으며, 그 유전자들을 각각 flo-a, flo-b, flo-c로 표시하였다. 10. flo-a 유전자는 IV번 연관군의 lg(liguleless ; 무엽설) 유전자와 5.76$\pm$1.72%(상반)로 연관되어 있었다.

• The Plant Pathology Journal
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• 제26권3호
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• pp.216-222
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• 2010

Fusarium proliferatum is the causal agent of stalk and root rot disease of maize, foot rot disease of rice, basal and root rot disease of onion and knife cut disease of sugarcane in Iran. In recent years, incidence and severity of these diseases have been increased in Iran. Fifty seven F. proliferatum single-spore isolates collected from diseased maize, rice, onion and sugarcane plants at different areas were used to study genetic diversity by determination of vegetative compatibility groups (VCGs). Chlorate-resistant nitrate non-utilizing (nit) mutants were recovered from selected isolates of F. proliferatum and used in complementation tests. All isolates in which both nit1 and NitM (or nit3) mutants were recovered, demonstrated self-compatibility. Vegetative compatibility tests by pairing nit mutants identified 30 VCGs among 57 isolates. Twenty-three isolates belonged to singlemember VCGs and the remaining 34 isolates, belonged to other seven multimember VCGs. Segregation of F. proliferatum isolates obtained from various area and host plants into different VCGs in Iran is reported for the first time. In this study, none of isolates obtained from rice complemented with any other isolates from onion and sugarcane and, non complementation occurred between onion and sugarcane isolates. Also, only one complementation occurred between one isolate of maize and one isolate of sugarcane and rice. Thus, a correlation between VCGs grouping and host preferences was founded. It is concluded that natural populations of F. proliferatum in Iran are probably genetically divergent and include isolates representing a potential risk for disease development.

• Molecules and Cells
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• 제42권10호
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• pp.711-720
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• 2019

Sink strength optimizes sucrose import, which is fundamental to support developing seed grains and increase crop yields, including those of rice (Oryza sativa). In this regard, little is known about the function of vacuolar invertase (VIN) in controlling sink strength and thereby seed size. Here, in rice we analyzed mutants of two VINs, OsVIN1 and OsVIN2, to examine their role during seed development. In a phenotypic analysis of the T-DNA insertion mutants, only the OsVIN2 mutant osvin2-1 exhibited reduced seed size and grain weight. Scanning electron microscopy analysis revealed that the small seed grains of osvin2-1 can be attributed to a reduction in spikelet size. A significant decrease in VIN activity and hexose level in the osvin2-1 spikelets interfered with spikelet growth. In addition, significant reduction in starch and increase in sucrose, which are characteristic features of reduced turnover and flux of sucrose due to impaired sink strength, were evident in the pre-storage stage of osvin2-1 developing grains. In situ hybridization analysis found that expression of OsVIN2 was predominant in the endocarp of developing grains. A genetically complemented line with a native genomic clone of OsVIN2 rescued reduced VIN activity and seed size. Two additional mutants, osvin2-2 and osvin2-3 generated by the CRISPR/Cas9 method, exhibited phenotypes similar to those of osvin2-1 in spikelet and seed size, VIN activity, and sugar metabolites. These results clearly demonstrate an important role of OsVIN2 as sink strength modulator that is critical for the maintenance of sucrose flux into developing seed grains.

• 방사선산업학회지
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• 제4권4호
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• pp.325-334
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• 2010

Transcriptional regulation in response to salt in mutant lines was investigated using oligonucleotide microarrays. In order to characterize the salt-responsive genes in rice, the expression profiles of transcripts that responded to salt-treatment were monitored using the microarrays. In the microarray analysis, among 37,299 reliable genes, 5,101, 2,758 and 2,277 genes were up-regulated by more than 2-fold using the salt treatment, while the numbers of down-regulated genes were 4,619, 3,234, and 1,878 in the WT, ST-495, and ST-532, respectively. From genotype changes induced by gamma ray mutagenesis, 3,345 and 2,397 genes were up-regulated, while 2,745 and 2,075 genes were down-regulated more than 2-fold in the two untreated mutants lines compared with untreated WT, respectively. A total of 3,108 and 2,731 genes were up-regulated more than 2-fold, while 3,987 and 3,660 genes were down-regulated by more than 2-fold in the salt treatment of the two mutants lines compared with the salt treated WT, respectively. The expressions of membrane transporter genes such as OsAKT1, OsKUP, and OsNAC increased more severely in ST-495 and ST-532 than in the WT. The expressions of the proline accumulation related genes such as OsP5CS and OsP5CR were also markedly increased in the salt tolerant mutants when compared to the WT plant.

• Animal cells and systems
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• 제10권4호
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• pp.197-201
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• 2006

Diaminopimelate decarboxylase (DAPDC, EC 4.1.1.20) catalyzes the conversion of diaminopimelate into lysine (Lys), which is the last step in Lys biosynthetic pathway. The genes for DAPDC have been reported in many bacteria, and more recently in Arabidopsis. Here we report characterization of a gene for DAPDC from rice (OsDAPDC). Sequence analysis of a cDNA clone revealed a full-length open reading frame for OsDAPDC that encoded 490 amino acids, approximately 53.2 kDa protein. The OsDAPDC protein contains a consensus binding site for pyridoxal-5'-phosphate as a cofactor and has a sequence at the amino terminus that resembles a transit peptide for localization to plastids, similar to that of Arabidopsis. Single gene encoding DAPDC was found in chromosome II in rice. The predicted amino acid sequence of OsDAPDC is highly homologous to that of the enzymes for DAPDC encoded by lysA of many bacteria. Expression of OsDAPDC in lysA mutants of Escherichia coli shows that the gene is able to functionally complement the mutants. These results suggest that OsDAPDC encodes a protein for diaminopimelate decarboxylase in rice.

• Molecules and Cells
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• 제20권3호
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• pp.385-391
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• 2005

As a first step towards identifying genes involving in the signal transduction pathways mediating rice blast resistance, we isolated 3 mutants lines that showed enhanced susceptibility to rice blast KJ105 (91-033) from a T-DNA insertion library of the japonica rice cultivar, Hwayeong. Since none of the susceptible phenotypes co-segregated with the T-DNA insertion we adapted a map-based cloning strategy to isolate the gene(s) responsible for the enhanced susceptibility of the Hwayeong mutants. A genetic mapping population was produced by crossing the resistant wild type Hwayeong with the susceptible cultivar, Nagdong. Chi-square analysis of the $F_2$ segregating population indicated that resistance in Hwayeong was controlled by a single major gene that we tentatively named Pi-hy. Randomly selected susceptible plants in the $F_2$ population were used to build an initial map of Pi-hy. The SSLP marker RM2265 on chromosome 2 was closely linked to resistance. High resolution mapping using 105 $F_2$ plants revealed that the resistance gene was tightly linked, or identical, to Pib, a resistance gene with a nucleotide binding sequence and leucine-rich repeats (NB-LRR) previously isolated. Sequence analysis of the Pib locus amplified from three susceptible mutants revealed lesions within this gene, demonstrating that the Pi-hy gene is Pib. The Pib mutations in 1D-22-10-13, 1D-54-16-8, and 1C-143-16-1 were, respectively, a missense mutation in the conserved NB domain 3, a nonsense mutation in the 5th LRR, and a nonsense mutation in the C terminus following the LRRs that causes a small deletion of the C terminus. These findings provide evidence that NB domain 3 and the C terminus are required for full activity of the plant R gene. They also suggest that alterations of the resistance gene can cause major differences in pathogen specificity by affecting interactions with an avirulence factor.

• 한국미생물·생명공학회지
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• 제6권3호
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• pp.129-133
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• 1978

Trichoderma viride QM 9414의 생육을 저해하여 colony를 형성 시키고 배지에 첨가된 cellulose가 균이 분비한 cellulase에 의해서 분해되어 생긴 clear zone으로서 육안으로 cellulase 역가를 쉽게 측정할 수 있는 방법과 repressor를 첨가하여 constitutive mutant를 선별할 수 있는 방법에 관한 실험을 하였든 바 그 결과를 요약하면 다음과 같다. 1) Colony를 형성시키고 그 size를 제한 하는데는 deoxycholate를 배지에 0.15％ 첨가 하였을 때 가 가장 좋았다. 2) Saponin을 배지에 0.l％ 첨가하면 뚜렷한 clear zone을 형성하였다. 3) Constitutive mutant를 선별하기 위해서는 기질로 pulp를 사용하였을 경우 glucsoe를 1％ 첨가하거나 glycerol이나 cellobiose를 0.5％ 첨가하면 가능하다고 생각되며 기질로 pure cellulose를 사용하였을 경우는 그대로 배지에 spore를 plating하면 가능하다고 생각된다. 4) Cellulase 역가가 높은 균주는 clear zone의 크기로서 쉽게 분리할 수 있다.