The purpose of this study was to develop the best mushroom cultivation conditions and the combination of mushroom culture media in order for mushroom producers and consumers. To reach this target, we first investigated the genetic relationship and developed suitable conditions of mycelial growth in Hypsizygus marmoreus strains. One superior strain of H. marmoreus was selected from 124 strains using bag culture. One hundred and twenty four strains were genetically classified into four main groups using two Universal Rice primers, URP2R and URP17R. The studies on the effects of different temperature (17, 21, 25, 29, 33℃) showed that 25℃ is the best temperature for mycelial growth for almost all strains while at 33℃ most of mycelium stop growth. Finally, ten strains were selected according to the groups identified by their temperature requirements. The length of mycelial growth in PDA, MCM, GPYM, MEA and MYP were longer than those in Czapek Dox. The selected ten strains of H. marmoreus showed heavier dry weight of mycelia at pH 3.0∼7.0 than any other pH. Although it was not show distinct requirement of carbon and nitrogen sources for vegetative growth according to strains, mainly the mycelial growth of the ten selected strains were observed at media including xylan and yeast extract, peptone, tryptone, respectively. Moreover, higher C/N ratio was observed in higher dry weight of mycelia.
TGsol (Translational Genomics for Solanaceae)은 가지과 작물의 유전체 정보를 육종분야에서 활용할 수 있도록 해석하여 제공하는 특화된 웹 인터페이스이다(http://tgsol.seeders. co.kr). 가지과는 경제적으로 고부가가치 작물인 토마토, 감자, 고추를 포함한 3,000개 이상의 다양한 종을 포함하고 있다. 최근 감자 (Nature, 2011)와 토마토 (Nature, 2012)의 유전체 정보가 발표되었고, 고추 유전체는 국내팀에서 독자적으로 많은 진척을 보여 이를 육종에 활용하기 위한 방안이 필요하다. 또한 주요 육종 라인 혹은 중요 유전자원을 중심으로 resequencing이 예정되어 있어 활용에 대한 기대가 매우 높으나 대규모의 복잡한 유전체 정보를 접근할 수 있는 통로는 매우 제한적이다. 그래서 TGsol은 가지과 작물 표준유전체 그리고 resequencing 정보를 통합하여 분자육종 수요자의 요구에 친화적인 웹 인터페이스로 구축하여 기능적 데이터베이스를 제공하고자 한다. 특별히 유전체 정보와 형질관련 유용유전자를 제공함으로써 MAS (maker-assisted selection)와 MAB (marker-assisted backcrossing) 확립에 필요한 다양한 정보를 제공한다. MAS를 위한 목표 형질은 병저항성, 과실발달, 유용 대사산물이다. 또한 토마토, 감자, 고추 사이의 ortholog 정보를 제공함으로써 기존에 연구된 형질관련 유전자를 가지과내에서 적용할 수 있도록 지원한다. 이러한 결과는 유전체 정보와 육종간의 상호 소통 및 활용이 원활하도록 지원하는 가교역할을 수행할 것으로 기대한다.
Clubroot, caused by a soil borne fungus Plasmodiophora brassicae Woronin, is a common disease of cabbages and other plants belonging to the genus Brassica, which is the most extensively cultivated vegetable crops worldwide. This present study was to evaluate the utilization possibility of SNP primers, which we designated as molecular markers linked to disease resistance based on B. rapa genome, it may be possible to apply them to B. oleracea, and to survey SNP marker related to clubroot resistance of cabbages. In total, 425 SNP markers can be applied to B. oleracea were selected from 8,000 SNP markers based on B. rapa genome linked to disease resistance. New 123 SNP markers of them were designed to be analysed to High Resolution Melt (HRM), and tested for clubroot resistance using 6 cabbage varieties, including 3 clubroot resistances (YR Chunrok, YR Dongjanggun, and Grandmart Cabbage) and 3 susceptibilities (Chungam-45, Bogam-1, and Junggam-21). Of them, 118 SNP primers amplified cabbage genomic DNA using HRM analysis, suggesting that it is possible to apply SNP markers based on B. rapa genome to B. oleracea. A total of 4 candidate SNP markers related to clubroot resistance were detected at 80.2℃ of melt temperature in BRS6, 79.2℃ in BRS18, 82.2℃ in BRS79, and at 84.4℃ in BRS114, respectively. These results provide valuable information that can be used for the utilization of the genus Brassica genome study and breeding for clubroot resistance in cabbages.
The rice blast disease caused by Magnaporthe oryzae (M. oryzae) is one of the most serious diseases of cultivated rice (Oryza sativa L.) in most rice-growing regions of the world. In order to investigate early responsible genes in rice in response to M. oryzae, we analyzed transcriptomics analysis using 300 K tilling microarray chip. The quality of RNA samples was initially validated by 4 defense related genes and phytoalexins measurement using RT-PCR and HPLC, respectively, which are well known defense markers. We determined that accumulation of 608 genes showed statistically significant changes in the level of transcription (>2 fold change, P<0.05). Among them, 261 genes were more up-regulated in incompatible interaction than that of compatible one. We further analyzed GO enrichment analysis of the 41 and 231 which were 2 fold up-regulated genes at 12h and 48h in incompatible interaction, respectively, using Rice Oligo nucleotide Array Database (http://ricearray.org). Furthermore, MapMan analysis (http://mapman.gabipd.org/) revealed that 21 and 85 genes including 18 receptor-like genes which were more induced in incompatible interaction compared to compatible interaction were found to be involved in biotic stress. Thus, this study suggests that early inducible genes including receptor-like protein kinases in incompatible interaction may play a key role in disease resistance against M. oryzae attacks.
R genes are a key component of genetic interactions between plants and biotrophic bacteria and are known to regulate resistance against bacterial invasion. The most common R proteins contain a nucleotide-binding site and a leucine-rich repeat (NBS-LRR) domain. Some soybean NBS-LRR genes have also been reported to function in disease resistance. A total of 319 genes were determined to be putative NBS-LRR genes in the soybean genome. The number of NBS-LRR genes on each chromosome was highly correlated with the number of disease resistance QTL in the 2-Mb flanking regions of NBS-LRR genes. In addition, the recently duplicated regions contained duplicated NBS-LRR genes and duplicated disease resistance QTL, and possessed either an uneven or even number of NBS-LRR genes on each side. The significant difference in NBS-LRR gene expression between a resistant near-isogenic line (NIL) and a susceptible NIL after inoculation of Xanthomonas axonopodis pv. glycines supports the conjecture that NBS-LRR genes have disease resistance functions in the soybean genome. The number of NBS-LRR genes and disease resistance QTL in the 2-Mb flanking regions of each chromosome was significantly correlated, and several recently duplicated regions that contain NBS-LRR genes harbored disease resistance QTL for both sides. In addition, NBS-LRR gene expression was significantly different between the BLP-resistant NIL and the BLP-susceptible NIL in response to bacterial infection. From these observations, NBS-LRR genes are suggested to contribute to disease resistance in soybean. Moreover, we propose models for how NBS-LRR genes were duplicated, and apply Ks values for each NBS-LRR gene cluster.
To produce abiotic stress resistant transgenic cucumber, the cotyledonary node explants of cucumber (c.v. Eunsung) were inoculated with A. tumefaciens strain EHA105 containing the binary vector (pPZP211) carrying Nit gene. The 491 explants inoculated with bacterium solution for 30 min were maintained on 50 mg/L paromomycin contained shoot induction (SI) medium for first 2 weeks and then subcultured on 100 mg/L paromomycin to obtain transgenic adventitious shoots for 4 x 14 days. So far, 5 plant were selected, and then acclimated in soil. Of them, 3 transgenic plants with Nit gene were confirmed by Southern blot analysis.
Microsatellite is one of the most suitable markers for variety identification as it has great discrimination power for varieties with narrow genetic variation. The relationship between marker genotypes and 58 melon varieties was analyzed. Of the 400 pairs of simple sequence repeat (SSR) primers screened against 11 melon varieties. 28 pairs showed highly polymorphism on the basis of PIC (Polymorphism Information Contents) value. These markers were applied to constructing DNA profile data base of 58 major melon varieties through multiplex PCR and fluorescence based automatic detection system. A total of 111 polymorphic amplified fragments were obtained by using 28 SSR markers. The average of PIC value was 0.643 ranging from 0.401 to 0.846. One hundred eleven SSR loci were used to calculate Jaccard’s distance coefficients for UPGMA cluster analysis. A clustering group of varieties, based on the results of SSR analysis, were categorized into plant shape and fruit type. Almost all of the varieties were discriminated by marker genotypes. This information may be used to select comparative variety through comparison of genetic relationship between existing variety and candidate varieties in distinctive tests and protection of plant breeders’ intellectual property rights through variety identification.
In plants, the Dof (DNA binding with One Finger) proteins are plant-specific transcription factors with a particular class of zinc-finger DNA-binding domain. The Dof genes have been predicted 30 different Dof genes in the rice Oryza sativa genome by phylogenetic analysis. The mostly Dof proteins contain a conserved region of 50 amino acids with a C2-C2 zinc finger motifs that binds a cis-regulatory element sequence 5’-T/AAAAG-3’. We found that a member of the DOF transcription factor family, Dof1 gene of rice, was expressed to wound from Ds insertion mutant population. Sequencing of the flanking regions of the transposon insertion site indicated that the gene-trap had been inserted near the front of the second exon of OsDof1 gene in chromosome 7. Genomic southern analysis revealed that mutant line contained a single copy of Ds gene trap. The Ds tagged rice mutant line, OsDof1::Ds, wound-inducible GUS expression was identified. To analyze the cis-acting elements, we constructed fusion genes with the OsDof1 promoter fused to the β-glucuronidase (GUS) reporter gene and transformed Arabidopsis and rice plants with these constructs. Wound-induced GUS expression was observed in the leaves of transgenic OsDof1::GUS rice and Arabidospsis plants. These results showed that, OsDof1 protein might be involved in stress responses and growth regulation in plant, might plays a role as a transcription regulator in stress response signal transduction pathways of plant.
The present study is designed to know the effect of cultivative environment and conditions, locational genetic variation on the content and components of seed isoflavones soybean genotype. The concentration of isoflavones showed difference between three different seeding time. Specially, Somyung of normal seeding (6. 21) revealed the highest concentrations (7657.0 ㎍ g-1) than other soybean seeds. On the other hand, Cheongjakong and Taekwangkong of early seeding (5. 25) showed the lower concentrations (1518.6 ㎍ g-1, 2524.0 ㎍ g-1) than other soybeans. For different regions, Taekwangkong of Yeoju region had the highest total isoflavone concentrations (3227.5㎍ g-1) than other cultivation regions. While, Taekwangkong of Miryang region (619.5 ㎍ g-1) revealed lower concentrations than other regions. And, Cheongjakong of Yeoju regions also revealed higher concentrations (3140.1 ㎍ g-1) than other regions. On the other hand, Cheongjakong of Miryang (1619.4 ㎍ g-1) showed lower concentrations of total isoflavone concentrations. In our present study, the isoflavone concentration of soybean was influenced by seeding and cultivation regions.
Rice transformation method using A. tumefaciens has already been widely used to generate transgenic plants, the transformation rate is still low in most Korean elite cultivars. We made several modifications of the standard protocol especially in the co-cultivation step to improve the efficiency of the rice transformation. The co-culture medium was modified by the addition of three antioxidant compounds (10.5㎎/ℓ L-cysteine, 1mM sodium thiosulfate, 1mM dithiothreitol) and of Agrobacterium growth-inhibiting agent (5㎎/ℓ silver nitrate). Co-cultivation temperature (23. 5℃ for 1 day, 26.5℃ for 6 days) and duration (7 days) were also changed. The plasmid of pMJC-GB-GUS carrying the GUS reporter gene and the bar gene as the selectable marker was used to evaluate the efficiency of the transformation. After co-cultivation, a high level of GUS gene expression was observed in calli treated with the modified method. It is likely that those newly added compounds helped to minimize the damage due to oxidative bursts during plant cell-Agrobacterium interaction and to prevent necrosis of rice cells. And the transformation rate under the modified method was also remarkably increased approximately 8-fold in Heungnambyeo and 2-fold in Ilmibyeo as compared to the corresponding standard method. Furthermore, we could produce the transgenic plants stably from Ilpumbyeo which is a high-quality rice but its transformation rate is extremely low. Transformation and the copy number of transgenes were confirmed by PCR, bar strip and Southern blot analysis. The improved method would attribute reducing the effort and the time required to produce a large number of transgenic rice plants.
Rice transformation method using A. tumefaciens has already been widely used to generate transgenic plants, the transformation rate is still low in most Korean elite cultivars. We made several modifications of the standard protocol especially in the co-cultivation step to improve the efficiency of the rice transformation. The co-culture medium was modified by the addition of three antioxidant compounds (10.5㎎/ℓ L-cysteine, 1mM sodium thiosulfate, 1mM dithiothreitol) and of Agrobacterium growth-inhibiting agent (5㎎/ℓ silver nitrate). Co-cultivation temperature (23. 5℃ for 1 day, 26.5℃ for 6 days) and duration (7 days) were also changed. The plasmid of pMJC-GB-GUS carrying the GUS reporter gene and the bar gene as the selectable marker was used to evaluate the efficiency of the transformation. After co-cultivation, a high level of GUS gene expression was observed in calli treated with the modified method. It is likely that those newly added compounds helped to minimize the damage due to oxidative bursts during plant cell-Agrobacterium interaction and to prevent necrosis of rice cells. And the transformation rate under the modified method was also remarkably increased approximately 8-fold in Heungnambyeo and 2-fold in Ilmibyeo as compared to the corresponding standard method. Furthermore, we could produce the transgenic plants stably from Ilpumbyeo which is a high-quality rice but its transformation rate is extremely low. Transformation and the copy number of transgenes were confirmed by PCR, bar strip and Southern blot analysis. The improved method would attribute reducing the effort and the time required to produce a large number of transgenic rice plants.
A cDNA clone encoding CBL-interacting protein kinase 1 (CIPK1) was isolated from Chinese cabbage seedlings. The gene, BrCIPK1 consisted of 1,982 bp long with 216 bp of the 5’-untranslated region (UTR), 1,509 bp of the coding region and 257 bp of the 3’-UTR. It is highly conserved CBL-interacting module with absolutely conserved domain among the 15 amino acid NAF domain of the 15 related genes. Southern blot analysis showed a single copy number. BrCIPK1 gene was localized in the cytoplasm and peripheral region in the plant cell which is highly expressed in seedling of rice and in the shoot and pistil of Arabidopsis. Analyses of gene expression on Ubi-1::BrCIPK1 rice lines was differentially accumulated by cold, salinity and drought, indicating its biological roles in the multiple stress response pathways in plants. Further, the expression of BrCIPK1 is hijacked by rice calcineurin-B-like protein (OsCBL5). Moreover, mRNA expression of P5CS1, a gene responsible for proline biosynthesis is regulated by the BrCIPK1 during abiotic stresses resulting to improved accumulation of proline. The interaction of BrCIPK1 with OsCBL5 along with the regulation of P5CS1 explained the enhanced tolerance of transgenic rice. This gene could be used in the development of rice varieties with enhanced tolerance to abiotic stresses.
The surveying of binding affinity between a particular transcription factor and DNA motifs is important in order to understand the developmental specific gene expression and regulatory networks of an organism. The microarray-based technologies (protein-binding microarrays; PBMs) provide useful predictions for understanding the transcriptional regulatory code in a genome-wide manner. The PBM was designed in such a way that target probes were synthesized as quadruples of all possible 9-mer combinations, named Q9-UPBM. Also, we developed rice promoter PBM (RPBM) using 19,480 rice promoter sequences containing 40 bp long probe with overlapping 20 bp (cover 1kb from 5’ upstream). We applied RISBZ1 protein, an endosperm specific basic leucine zipper transcription factor, to compare binding site specificities between Q9-UPBM and RPBM and find directly regulated promoter regions through the RPBM. Several cis-elements; Prolamin box (TGTAAAG), GCN4 motif (TGA(G/C)TCA), AACA motif (AACAAAA), and ACGT motif, are highly conserved in the promoters of cereal seed storage protein genes, and play a central role in controlling endosperm specific expression during seed maturation. Characterization of cis-elements and TFs has been performed on many storage protein genes of several crop plants, but the mechanisms are still poorly understood. Two chips provide RISBZ1 could bind to ACGT motif such as a CCACGTCA site and GGATGAC site as well as GCN4 motif known binding site. In RPBM binding affinity to CCACGTCA was highly significant, compared to GGATGAC site. The difference might be caused by the biased presence of specific promoter rather than Q9-UPBM. Also our results will provide direct insight into the importance of combinatorial interplay between cis-elements in regulating the expression of seed storage protein genes.
Bacillus thuringiensis (Bt) crystal protein genes encode insecticidal δ-endotoxins that are widely used for the development of insect-resistant crops. Common soybean is a crop of economic and nutritious importance in many parts of the world. Korea soybean variety Kwangan was transformed with Bacillus thuringiensis (Bt) crystal protein genes. These genes were transformed into Kwangan using highly efficient soybean transformation system. Transgenic plants with Bt crystal protein genes were confirmed for gene introduction and their expression using PCR, real-time PCR and RT-PCR. Currently, the confirmation of stable gene introduction with Bt genes is also performing by southern blot analysis and physiology test and agronomic characters are investigating.
Cry1Ac protein is known as one of toxin crystal proteins synthesized from Bacillus thuringenesis that plays a critical role for the insect resistance. Recently, cry1Ac genes have introduced into many plants in general and soybean as well. However, the gene expression of cry1Ac genes in transgenic plants remains low that need to be improved. Several mutations we reintroduced into the cry1Ac genes in order to enhance the insecticidal effect. In this study, the cry1Ac with mutant #2, #11 and #16 were transformed into Kwangan, a Korean soybean variety by using the “half-seed” method. The plant lets carrying modified cry1Ac genes were primarily selected on media containing Phosphinothricine (PPT), a bar selective agent and Basta leaf painting. Then, the presence of introduced genes in T0 plants and the gene expression were investigated by PCR, RT-PCR and Real-time PCR. PCR and RT-PCR analysis showed expression of bar and cry1Ac genes from tested transgenic soybean plants. The number of copy of bar gene ranged from 1 to 3 by Real-time PCR analysis. These results provided a fundamental back ground for our further experiments: Confirmation of the gene expression by Southern blot and identification of the function of modified cry1Ac by insect bioessays.
GM 벼의 환경위해성 평가를 위해 레스베라트롤 생합성 벼인 익산515호, 익산526호와 동진벼를 수원, 익산, 밀양 등 3개 지역의 GMO 격리포장에서 재배하여 지역별 농업적 특성과 레스베라트롤 함량을 조사하였다. 익산과 밀양에서 재배된 3계통 모두 생육특성 및 수량 등 농업적 특성이 비슷한 결과를 나타내었으나, 수원지역에서 재배된 시험계통 모두 익산 대비 출수가 지연되고, 키는 12㎝ 이상 작고, 수수, 수당립수, 등숙률이 낮아져 익산지역과 비교하여 쌀 수량이 약 12% 정도 감수되었다. 하지만, 대조구인 동진벼 대비 익산515호, 익산526호의 수량지수는 익산, 수원, 밀양 등 3개 지역 모두 동일한 경향을 나타내었다. 3개 지역에서 재배된 동진, 익산515호, 익산526호 등 3계통의 정조 시료를 백미로 조제한 후, UPLC를 이용하여 레스베라트롤 함량을 분석한 결과, 익산지역 대비 익산515호, 익산526호 계통의 레스베라트롤 함량은 수원, 밀양에서 생산된 2계통 모두 각각 약 30%, 25% 이상 함량이 증가하였다. 이는 레스베라트롤 합성 유전자가 온도, 일조량 등 재배환경에 따라 유전자 발현량이 조절되어 최종산물인 레스베라트롤 생합성에 영향을 주는 것으로 확인되었다.
Disease is one of the significant factors to damage for the crop productivity, including rice. Although there are many methods to avoid from several diseases such as chemical pesticides and biological treatments, it has been appreciated that the most economical and environmentally effective method of disease control is application of resistance genes. A survey (Dardick & Ronald, 2006) reported that plant kinome has a small number of non-RD kinase (nRDK) (4-29% of total kinase), all known or predicted pattern recognition receptors (PRRs) fall into the class. We here introduce a strategy to identify rice resistance genes that are probably encoding PRRs. We selected 130 nRDK genes by combinational analysis of QTL and bioinformatics, 61 of rice mutant lines of 130 candidates inoculated by Xanthomonas oryzae pv. oryzae (Xoo) and Magnaporthe grisea. (M. grisea), and disease progression was monitored. Lesion lengths of the activation mutant lines for nRDK-08 and nRDK-18 genes reduced more than 34% compared to wild type of rice (Dongjin) and other mutant lines. The nRDK-03 and nRDK-17 gene activation rice line had remarkably smaller lesion lengths by M. grisea infection. Our results suggest that a reverse genetic approach using bioinformatics and T-DNA tagging system successfully identified nRDK genes conferring a resistance against Xoo and M. grisea.
This study investigated the functional substances, including phenolics, tocopherol compounds in rice (Onnuri, Nampyeong, Dongjin, Deuraechan, Hopum, Iksan-515, Iksan-526) that were cultivated in Iksan region. Specially, Iksan-515 and Iksan-526 were produced resveratrol rice. Resveratrol is health-beneficial compound with strong antioxidant activity. Iksan-515 and Iksan-526 were contained resveratrol compound while, other rice varieties did not showed resveratrol. The total phenolic compounds showed similar concentrations in 7 rice varieties. Especially, Iksan-526 revealed 273.3 ㎍ g-1 and Iksan-515 was 264.8 ㎍ g-1. In this study, we analyzed vitamin E contents of 7 varieties of rice. The total tocopherols revealed 418.3 ㎍ g-1 in Iksan-526, 401.2 ㎍ g-1 in Iksan-515 and 413.3 ㎍ g-1 in Hopum. We determined 4 tocopherols (α-, β-, γ-, δ-) and 4 tocotrienols (α-, β-, γ-, δ-). Among the all samples, β-tocotrienol showed higher average concentrations (101.3 ㎍ g-1) than other compounds and, γ-tocotrienol revealed second higher concentration (94.5 ㎍ g-1). On the other hand, δ-tocopherol was not detected in all rice samples.
This study was carried out to determine the contents of antioxidant activity of colored rice lines which derived from a mutant of MGI079 induced by MGI079 tissue culture in rice (Oryza sativa L.). The antioxidant activities were evaluated by assaying polyphenol, flavonoid, DPPH free radical and color values of colored rice lines, respectively. Among 8 lines including Heugnam of colored rice, the 70% ethanol extracts decreased in the order of MGI079-2-1>MGI079-2-6>MGI079-2-R,Heugnam>MGI079-1-R>MGI079-2-1>MGI079, MGI079-1-1, MGI079-1-6. The rice lines of highest polyphenolic compound was MGI079-2-6 and the next were Heugnam, MGI079-2-R, MGI079-2-1, MGI079-1-R, MGI079-1-6, MGI079-1-1, MGI079, MGI079-OP-R with the order of higher content. The flavonoid was higher in order of MGI079-2-1, MGI079-2-6, Heugnam, MGI079-2-R, MGI079-1-R, MGI079-1-6, MGI079-OP-R, MGI079, MGI079-1-1. The DPPH free radical was higher in order of MGI079-2-1, MGI079-2-6, Heugnam, MGI079-1-6, MGI079-1-1, MGI079-2-R, MGI079-1-R, MGI079-OP-R, MGI079. For chromaticity, a negative correlation was exhibited between the color value and the 70% ethanol extracts, polyphenolic compound, flavonoid, DPPH free radical. The grain characters in brown rice of a mutant of MGI079 showed similar to that of a donor plant, MGI079. Whereas, chemical characteristics of brown rice in two colored rice lines(MGI079-2-1, MGI079-2-6) were lower in amylose and lipid contents and were higher four times in zinc that of a donor plant, MGI079. Two colored rice lines(MGI079-2-1, MGI079-2-6) showed relatively high antioxidative activity in every results of antioxidant activity tests.
The significance of genetic stability and bio-safety environment has been recently recognized by many GM plants. This study was to evaluate the GM stability of transgenic rice and to identify the environment variance. The GM rice of vitamin A -enriched rice and four check cultivars were analyzed the data on agronomic characters and principal component for 2009-2011 in large-GM crop field. Cultivation environment was conducted in the large-GM field and greenhouse to determine grain characters. In this experiment, there was no significant difference in agronomic characters between GM rice of vitamin A-enriched rice and a donor plant, Nagdong. Related to grain characters, grain appearance and physicochemical characteristics were similar to GM rice of vitamin A-enriched rice and a donor plant, Nagdong. However, grain appearance in GM rice of vitamin A-enriched rice showed to white core and white belly when GM rice of vitamin A-enriched rice was planted in greenhouse. The type and distribution of dominant weed species also were not different from GM rice of vitamin A-enriched rice and a donor plant, Nagdong. Additionally that of gene flow was not detected in dominant weed species by PCR analysis.