The maize genome is complex with exceeding the levels of intra-specific variation, repetitive DNA content, and allelic content observed between many species. Because of tremendous diversity and variants, maize is considered as a forefront crop development and estimation of molecular markers for agricultural trait in genetics and breeding. Using quantitative trait loci (QTL) and marker assisted breeding (MAS), molecular breeders are able to development of drought tolerance and grain yield in maize genotype. To study QTL congruency, a meta QTL analysis including results from eight-teen QTL publications for grain yield and drought tolerance were considered. Among them, we assembled 420 QTLs for abscisic acid (ABA) concentration, anthesis silking interval (ASI), days to flower, days to silk, ear number, kernel number, grain number and grain yields, involved in drought tolerance and grain yield. The meta QTL analysis revealed significant evidence for linkage of these traits to 39 different segments as candidates regions on maize genome. A total of 571 marker was selected as QTL or integrated QTL markers for narrowing down the QTL region into specific functionally relevant candidates. The results of meta QTL analysis helped to refine the genomic regions of agricultural traits, interest described and provided the closest flanking markers.
In here, we screened drought tolerant varieties with modified leaf water loss rate assay and visual drought tolerant phenotype in the greenhouse conditions with more than 800 varieties. Among these varieties, Samgang, Gumei4 and Apo showed the lowest of leaf water loss rate and strong drought tolerant phenotype. To identify drought QTLs with Samgang variety, we developed the doubled-haploid (DH) population consist of 101 lines derived from a cross the drought tolerant cultivar Samgang and the drought sensitive cultivar Nagdong. To score the drought phenotype degrees of this population, we withheld water for 6 weeks and treated the watering for 7 days. After watering, visual phenotype was observed 1 to 9 degree according to the standard evaluation system for rice, IRRI. Drought sensitive parent Nagdong was almost died and was scored as 9 degree, while tolerant parent Samgang showed slightly leaf tip drying phenotype and was scored as 3 degree in our experimental conditions. Three main QTLs were detected on chromosome 2, 6, and 11 with this visual phenotype. We also measured relative water contend of these population under drought stress conditions, and got one main QTL on chromosome 11. The QTL loci on chromosome 11 with flanking markers RM26755-RM287 has a function for visual phenotype and relative water content under drought conditions.
Cultivated tomato (Solanum lycopersicum L.) is an economically important vegetable and has a narrow genetic base due to intensive human selection through domestication and breeding. The low level of genetic variation between cultivated tomatoes has made it difficult to develop molecular markers for elite breeding lines. Recently, genome-wide 145,695 InDels were identified from in silico analysis of two tomato genome sequences, Heinz 1706 (S. lycoperiscum) and LA1589 (S. pimpnellifolium). Of these, 2,272 InDels were validated and 717 InDels showed polymorphism in cultivated tomatoes. In the present study, we selected 48 out of 717 InDels based on PIC value (> 0.3) and size (> 10 bp) to develop a DNA database for commercial tomato cultivars. We also used an additional set of 28 InDels that have been previously reported. These markers were distributed across 11 chromosomes with an average of 6.6 markers. A total of 48 F1 hybrid cultivars were collected from 20 seed companies and a subset of eight cultivars were used to test polymorphism of the InDel markers. The 37 InDel markers were polymorphic in these cultivars and were used to genotype additional 40 cultivars. Genetic distances and relationships between cultivars were assessed using the InDel genotypes of 48 cultivars. This analysis revealed that the InDel markers detected genetic variations to identify 46 cultivars. Our results demonstrate that the InDel markers will be a useful resource to construct a DNA database for tomato cultivars and to protect tomato breeder’s rights via variety identification.
Proline (Pro) accumulation is a common physiological reaction in response to abiotic stresses in many plants. Accumulation of Pro is believed to play the important role in protecting cellular components from dehydrating effects due to such stresses. The study was performed to investigate the relationship between cold hardiness and Pro content or expression of related genes in peach cultivars during a constant experimental deacclimation. Changes in cold hardiness were determined using electrolyte leakage method in the shoots of 10 peach cultivars (Prunus persica ‘Aikawanakajima’, ‘Chiyomaru’, ‘Daewol’, ‘Janghowon Hwangdo’, ‘Kiraranokiwami’, ‘Mihong’, ‘Misshong’, ‘Soomee’, ‘Suhong’, and ‘Sun Gold’). Pro content was analyzed using the ninhydrin method and related gene expressions were examined using quantitative real-time RT-PCR. While cold hardiness of 10 peach cultivars decreased, Pro contents of those increased during the deacclimation. Notably, at the same time, expression of P5CS (Δ1-pyrroline-5-carboxylatesynthase) decreased in 10 peach cultivars, whereas expressions of P5CR (Δ1-pyrroline-5-carboxylatereductase) and OAT (ornithine-δ-aminotransferase) increased. Our results demonstrate that Pro responds positively to higher temperature in the shoots of 10 peach cultivars and expression of both P5CS and P5CR genes could show contrasting patterns during the deacclimation. Furthermore, our results suggest that ornithine pathway, which has been suggested to be important during seedling development, could serve as an alternative pathway in Pro synthesis process during the deacclimation in peach.
Root-knot nematode, Meloidogyne incognita is a virulent pest of solanaceaous crops worldwide. The M. incognita resistance gene Me7 derived from Capsicum annuum CM334, is located on chromosome 9. In the present study, an F2 population derived from a cross between ECW03R and CM334 was used to locate the Me7 gene. An F2 population was inoculated using approximately 1,000 second-stage juveniles per individual plant. Phenotype screening was done 45 days after inoculation by using gall index system. The phenotype study of 503 F2 individual showed 391 resistant and 112 susceptible plants. The 3:1 phenotypic ratio confirmed that resistance phenotype is controlled by a single dominant gene. Previously reported two markers were tested to reveal the linkage of markers to phenotype. Two markers, CAPS_F4R4 and SCAR_PM6a were located at 4.3 and 2.7 cM from the resistance gene, respectively. Additional SNP markers were developed using CM334 reference genome information to narrow down the position of the gene, but no closer markers could be developed due to errors of DNA sequence assembly. The closest marker was positioned on telomere of the chromosome 9 long arm, where tens of other NB-LRR genes are clustered. NB-LRR genes are being used as candidates to identify the Me7 gene.
알스트로메리아의 환경위해성 평가를 하기 위한 생물학적 특성평가 방법을 제시하고자 본 연구를 수행하였다. 환경위해성 평가를 위해 전남대학교 격리 포장 및 격리온실에서의 원예 형질 표현형 지표비교, 격리 포장에서의 번식성과 월동성, 무성번식을 통한 차세대의 표현형적 후대 안정성 등 4항목의 생물학적 특성 조사를 하였으며, 또한 알스트로메리아의 실질적 동등성 평가를 하는 데 필요한 적정 실험 개체 수를 제시하고자 하였다. 격리 포장 및 격리 온실에서 원예형질 표현형 지표 비교 조사 결과 줄기 굵기, 잎의 길이와 너비, 꽃차례 분지 길이 및 꽃의 꽃자루 길이 등 5항목에서 포장보다 온실에서 크거나 굵은 차이를 보였다. 자연환경에서 번식성 조사방법은 알스트로메리아 rhizome의 개수를 조사한 결과 7.0에서 10.2개로 계통에 따라 차이가 나타났으며, 월동성은 토양 표면에서의 깊이를 10cm, 20cm, 30cm로 다르게 하여 월동 후 생존력을 조사한 결과 계통에 따라 10∼75%로 조사되었다. 무성번식을 통한 차세대의 표현형적 후대 안정성 조사에서는 양적 형질 6가지를 조사하였으며, 씨방에서 안토시아닌의 유무를 조사한 결과 ‘씨엔알스호프’(품종등록번호 5192)는 안토시아닌이 있고, ‘파이네세’는 안토시아닌이 없었던 차이를 제외하고 다른 항목에서는 차이를 발견할 수 없었다. 실질적 동등성 평가를 하는데 필요한 적정 실험 개체 수는 전남대학교 온실에서 표준재배법에 따라 재배된 알스트로메리아 96주를 대상으로 하였으며 양적 형질 9항목의 특성을 실측하여, 각 조사항목의 실측치를 로그함수에 의한 비선형회귀모형을 사용하여 분석하여 기울기 값이 0.03이 되는 점을 정하였으며, 조사결과 실질적 동등성 평가를 하기 위한 개체 수는 항목에 따라 13주 이상에서 60주 이상으로 조사되었다.
Tomato fruit color, which is the most visible characteristic among the other fruit traits, is considered to have a substantial influence on consumers. The pink-colored tomatoes with high soluble solids content are considerably preferred especially in Asia compared to the other colors. Generally the pink fruit trait of tomatoes is easily determined by visual examination of intact fruit, however, it is technically determined by the characteristic of the fruit peel. The pink trait is regulated by variations of the SlMYB12(y) gene located on chromosome 1, which controls the accumulation of the naringenin chalcone, which comprises a large proportion of flavonoids. In this study, we developed a derived Cleaved Amplified Polymorphic Sequences (dCAPS) marker and a sequence characterized amplified regions (SCAR) marker in order to discriminate of pink/non-pinktomatoes in the domestic breeding lines. Quantitative RT-PCR analysis indicated that the SlMYB gene is highly expressed in non-pink fruit peel, whereas the expression is significantly lowered in the pink fruit peel. These gene based markers are expected to enhance the efficiency and accuracy of selection pink-tomatoes in tomato breeding programs.
The β-carotene biofortified transgenic rice was developed by transforming rice cv. Nakdongbyeo with phytoene synthase (Psy) and carotene desaturase (Crt I) genes isolated from Capsicum and Pantoea. The aim of this study was to perform molecular characterization of rice transformants of T5-T7 generation harboring Psy and Ctr I genes driven by endosperm specific globulin promoter for biosafety evaluation of β-carotene biofortified transgenic rice. The structure and sequence of T-DNA in the transformation vector and the insertion sites, flanking sequences and generational stability of inserted T-DNA in transgenic rice lines were analyzed. The transformation vector consisted of right border, MAR gene, carotenogenic genes unit, herbicide resistance selectable marker unit, MAR gene and left border in sequential order. T-DNA was introduced at the position of 30,363,938-30,363,973 bp of chromosome No. 2 by adaptor-ligation PCR. Stable integration of T-DNA and stable expression of bar gene was confirmed in T5 to T7 generations. It was also confirmed that the backbone DNA of transformation vector containing antibacterial gene was not present in the genome of β-carotene biofortified transgenic rice. HPLC analysis confirmed that carotenoids were consistently detected through T5-T7 generations.
Genetically modified (GM) crops have never been cultivated commercially in Korea, it is necessary for a thorough assessment of the risks associated with their environmental release. We determined the frequency of pollen mediated gene flow from disease resistant GM rice (OsCK1) to non-GM rice (Nagdongbyeo) and weedy rice (R55). A total of 449,711 or 164,604 seeds were collected from non-GM and weedy rice, respectively which were planted around OsCK1. Resistance of the hybrids was determined by repeated spraying of herbicide and DNA analysis using specific primer to confirm hybrids. Though non-GM rice and weedy rice have similar flowering time, the hybrids were found only in non-GM rice and out-crossing ranged from 0.018% at 0.3 m to 0.013% at 0.6 m. All of hybrids were located within 0.6 m distance from the GM rice plot in southerly direction. The meteorological factors including temperature and relative humidity during flowering time were found to be the most important factors for determining rice out-crossing. It should be considered many factors like the local weather condition and flowering time to set up the safety management policy to prevent pollen mediated gene flow between GM and conventional crop.
Proline has been shown to accumulate in plant under various type of stresses. In our previous study, changes in cold hardiness and proline content showed contrasting patterns during a constant deacclimation. This study was performed to investigate the proline accumulation and related gene expression in response to repeated deacclimation and reacclimation in peach cultivar ‘Daewol’. Proline content was analyzed using the ninhydrin method and related gene expressions were examined using quantitative real-time RT-PCR. Proline contents of ‘Daewol’ increased during the repeated deacclimation treatments. Interestingly, during the twice deacclimation, expressions of P5CS (Δ1-pyrroline-5-carboxylatesynthase) constantly decreased, whereas expressions of P5CR (Δ1-pyrroline-5-carboxylatereductase) increased. Expressions of OAT (ornithine-δ-aminotransferase) indicated up- and down- pattern in response to repeated deacclimation and reacclimation. Our results indicated that proline responds positively to higher temperature in the shoots of peach cultivar ‘Daewol’ and expressions of both P5CS and P5CR genes could show contrasting patterns during the deacclimation. Moreover, our results suggest that ornithine pathway could serve as an alternative pathway in proline synthesis process during deacclimation in peach.
Tomato spotted wilt virus (TSWV) causes one of the most destructive viral diseases that threaten tomato (Solanum lycopersicum) worldwide. So far, eight TSWV resistance genes, Sw1a, Sw1b, sw2, sw3, sw4, Sw-5b, Sw-6, and Sw-7 have been identified and Sw-5b has been incorporated into tomato for prevention of TSWV. The objectives of this research are first to discover single nucleotide polymorphisms (SNPs) in Sw-5 alleles and then to develop SNP markers to distinguish resistant genotypes against TSWV for marker-assisted breeding in tomato. First, DNA sequences of Sw-5b alleles from both resistant and susceptible cultivars amplified using known Sw-5 gene-based marker was analyzed. The single functional SNP (G→A) was detected as non-synonymous substitution because this SNP causes change of arginine (Arg599) to glutamine (Gln599). Next, the primer pair for high resolution melting analysis (HRM) was designed around this SNP. To determine accuracy of this SNP marker to distinguish resistant Sw-5b genotypes against TSWV, genotypes of 32 commercial tomato cultivars were checked. The newly developed SNP marker could select six cultivars carrying resistant Sw-5b genotype, which was 100% correlated with genotypes based on the gene-based marker. These results indicate that the SNP maker developed in this study could be useful for better tracking resistance to TSWV in tomato breeding.
Leaf mold disease in tomato (Solanum lycopersicum) is caused by Cladosporium fulvum, a fungal leaf pathogen. One of effective ways to control leaf mold is to breed disease-resistant tomato cultivars. Cf-4 and Cf-9 resistance (R) genes encode proteins that carry a leucine rich repeat domain and are located in plasma membrane. They trigger hypersensitive response following recognition of corresponding Avr4 and Avr9 proteins of C. fulvum, respectively. Cf-4 and Cf-9 genes are originated from wild tomato species S. habrochaites and S. pimpinellifolium and have been introgressed into commercial tomato cultivars. These two highly homologous orthologs exist as a cluster with four highly homologous paralogs. Due to this reason, development of genetic markers to distinguish these two functional R genes from their orthologs and paralogs is difficult. In this study, we tried to develop single-nucleotide polymorphism (SNP) markers to select tomato cultivars carrying resistant Cf-9 genotype. The genomic sequences of resistant Cf-4 and Cf-9 alleles, susceptible cf-9 alleles, and their paralogs were obtained from the GenBank database, and two functional SNPs causing non-synonymous substitution were found among them. Based on two SNPs, the Cf-9_2-SNP-F/R primer set for high resolution melting (HRM) analysis was developed. HRM analysis with this primer set could successfully distinguish tomato cultivars carrying resistant Cf-9 allele among 30 commercial tomato cultivars, which were characterized with the gene-based marker. These indicate that the SNP marker developed in this study is useful to trace Cf-9 genotype efficiently in marker-assisted selection in tomato.
Molecular characterization of crops improved through biotechnology has traditionally been conducted using Southern blot analysis which has been used to determine T-DNA copy number, the presence or absence of backbone (sequence outside of the T-DNA) and to demonstrate generational stability of the T-DNA insert. The advancement of high-throughput DNA sequencing (HTS) technology allows efficient characterization of the transgene incorportated into the genome of the plant by rapidly sequencing the entire plant genome. By combining NGS (Next Generation Sequencing) technologies with bioinformatic methods that identify the T-DNA insert derived from the plasmid vector and genome-T-DNA junction sequences, it has been shown that conclusions equivalent to those of a Southern blot are readily obtained. NGS is done at sufficient coverage depth (>75x) across the entire genome. By mapping the sequence reads to the plasmid vector, and identifying the number of unique junctions, we can confirm insert number, copy number, absence of backbone, across multiple generations. With the widespread availability of NGS and steadily decreasing costs it is likely that academia and industry will fully transition to NGS-based molecular characterizations in the near future.
Understanding how crops interact with their environments is increasingly important in breeding program, especially in light of highly anticipated climate changes. A total of 150 recombinant inbred lines (RILs) of F12 generation derived from Dasanbyeo (Indica) x TR22183 (Japonica) were evaluated at Suwon 2010, Shanghai 2010, IRRI 2010 wet season, Suwon 2011, Shanghai 2011, IRRI 2011 dry season, and IRRI 2011 wet season as a total of seven diverse environments. Traits evaluation included eight important agronomical traits such as days to heading (DTH), culm length (CL), panicle length (PL), panicle number per plant (PN), spikelet number per panicle (SN), spikelet fertility (SF), 100-grain weight (GW), and grain yield (GY). As a result of genotyping using 384-plex GoldenGate oligo pool assay (OPA) set (RiceOPA3.1), the linkage map for 235 SNP markers covering a total of 926.53 cM with an average interval of 4.01 cM was constructed and a total of 44 main-effect quantitative trait loci (QTL)s and 35 QTLs by environment interaction (QEI) were detected for all eight traits using single environment and multi-environments analysis, respectively. Of these, fourteen putative QTLs for DTH, CL, PN, SN, GW and GY found in single environment analysis had the similar position to QEI for those traits, suggesting that these same QTLs from both single-and multi-environments are major and stable for certain traits. To the best of our knowledge, 12 QTLs consisted of four QTLs for CL (qCL2, qCL8.1, qCL8.2, and qCL8.3), six QTLs for GW (qGW3.1, qGW3.2, qGW7, qGW8, qGW10.1, and qGW10.2), one QTL for GY (qGY3) and one for SF (qSF4) out of 44 QTLs obtained from single environment analysis were considered to be novel since no overlapping QTL was reported from previous studies. In addition, 12 out of 35 QTLs obtained from multi-environments analysis were also novel.
Copper (Cu) is an essential micronutrient required for growth and development of plants. But, at a high concentration in soil, copper acts as a major toxic element to plant cells due to its potential inhibitory effects against many physiological and biochemical processes. In this study, the morphological and physiological changes were observed in the leaf of sorghum plants treated with different concentrations (0, 100, and 150 μM) of Copper (Cu). The results linked to morphological changes that plants treated with Cu suffered reduction in growth and morphological changes. In the ion concentration investigation, the concentrations of Cu2+ increased, the concentration of others interacting ions (Zn2+, Ca2+, Mn2+, Fe2+) were changed dramatically. For proteome analysis, 2-D combined with MALDI-TOF-TOF mass spectrometry was performed. Two dimensional gels stained with silver staining, a total of 422 differential expressed proteins (≥ 2-fold) were identified using Progenesis SameSpot software. A total of 24 spots from Cu-induced sorghum leaf and 21 spots from Cu-induced sorghum root were analyzed by mass spectrometry. Out of 24 protein spots from Cu-stressed leaf, of which 16 protein spots were up-regulated and 8 protein spots were down-regulated whereas out of 21 protein spots, a total of 9 protein spots were up-regulated and 12 spots were down-regulated from Cu-stressed root. Taken together, these studies revealed the effects of heavy metal, Cu on the growth and physiological characteristics in sorghum seedlings and proteome investigation, hoping to provide references on the mechanism of heavy metal damaging plants.
Cadmium (Cd) pollution is thought to be one of the leading threat to the environment due to its high toxicity. However, the molecular responses induced by Cd have so far been grossly overlooked. This study examines the morpho-physiological alterations combined with proteome changes in leaves of Sorghum bicolor when exposed to Cd. Ten days old sorghum seedlings were exposed to different concentrations (0, 100, and 150 μM) of CdCl2 and a significant accumulation of Cd in the leaves was recorded by ICP analysis. Furthermore, the effects of Cd exposure on protein expression patterns in S. Bicolor was investigated by two-dimensional gel electrophoresis (2-DE) and the 2-DE profile of leaf proteins from both control and Cd-treated seedlings were compared quantitatively using Progenesis SameSpot software. Results lined to morphological changes that plants treated with Cd suffered reduction of growth. The concentration of Cd was markedly reversed by the Cd treatments, whereas the absorption degree of Cd was increased by the higher concentration of Cd by confocal microscopy. Using 2-DE method, a total of 33 differentially expressed protein spots were identified by MALDI-TOF-TOF mass spectrometry. Of those, 13 protein spots were significantly enhanced/reduced while 20 reduced under Cd treatment. The most of the up-regulated proteins are involved in oxidative response, glutathione and sulfur metabolism as well as the secondary metabolite biosynthesis. Collectively, our study provides insights into the integrated molecular mechanisms of early responses to Cd and growth and physiological characteristics of sorghum seedlings hoping to provide references on the mechanism of heavy metal damaging plants.
The roots of Platycodon grandiflorum are known as traditional medicine, has been extensively used since ancient times as a therapeutic to treat cold, cough and asthma in Korean traditional medications. This study was conducted in order to profile proteins from the hormone induced diploid and tetraploid roots using high throughput proteome approach. Two dimensional gels stained with CBB, a total of 64 differential expressed proteins were identified from the diploid root using image analysis by Progenesis SameSpot software. Out of total differential expressed spots, 20 differential expressed protein spots (≥ 2-fold) were analyzed using MALDI-TOF-TOF mass spectrometry whereas a total of 13 protein spots were up regulated and 7 protein spots were down-regulated. However, in the case of tetraploid root, a total of 78 differential expressed proteins were identified from tetraploid root of which a total of 28 differential expressed protein spots (≥ 2-fold) were analyzed by mass spectrometry whereas a total of 16 protein spots were up regulated and a total of 12 protein spots were down-regulated. However, proteins identified using iProClass databases revealed that the identified proteins from the explants were mainly associated with the nucleic acid binding, oxidoreductase activity, transporter activity and isomers activity. The exclusive protein profile may provide insight clues for better understanding the characteristics of proteins and metabolic activity in various explants of the economically important medicinal plant Platycodon grandiflorum.
The roots of Platycodon grandiflorum are massively used in traditional herbal medicine as a remedy for pulmonary disease and respiratory disorders. However, in spite of its potential medicinal significance, the molecular mechanism of its roots is still unknown. In the present study, high throughput proteome approach was conducted to profile proteins from 3, 4 and 5 months aged diploid and tetraploid roots of Platycodon grandiflorum. Two dimensional gels stained with CBB, a total of 68 differential expressed proteins were identified from diploid root out of 767 protein spots using image analysis by Progenesis SameSpot software. Out of total differential expressed spots, 29 differential expressed protein spots (≥ 2-fold) were analyzed using LTQ-FTICR MS whereas a total of 24 protein spots were up regulated and 5 protein spots were down-regulated. On the contrary, in the case of tetraploid root, a total of 86 differential expressed proteins were identified from tetraploid root out of 1033 protein spots of which a total of 39 differential expressed protein spots (≥ 2-fold) were analyzed using LTQ-FTICR MS whereas a total of 21 protein spots were up regulated and a total of 18 protein spots were down-regulated. It was revealed that the identified proteins from the explants were mainly associated with the nucleotide binding, oxidoreductase activity, transferase activity. In that way, the exclusive protein profile may provide insight clues for better understanding the characteristics of proteins and metabolic activity in various explants of the economically important medicinal plant Platycodon grandiflorum.
The β-carotene biofortified transgenic soybean was developed recently through Agrobacterium -mediated transformation using the recombinant PAC (Phytoene synthase-2A-Carotene desaturase) gene in Korean soybean (Glycine max L. cv. Kwangan). GM crops prior to use as food or release into the environment required risk assessments to environment and human health in Korea. Generally, transgenic plants containing a copy of T-DNA were used for stable expression of desirable trait gene in risk assessments. Also, information about integration site of T-DNA can be used to test the hypothesis that the inserted DNA does not trigger production of unintended transgenic proteins, or disrupt plant genes, which may cause the transgenic crop to be harmful. As these reasons, we selected four transgenic soybean lines expressing carotenoid biosynthesis genes with a copy of T-DNA by using Southern blot analysis, and analyzed the integration sites of their T-DNA by using flanking sequence analysis. The results showed that, T-DNA of three transgenic soybean lines (7-1-1-1, 9-1-2, 10-10-1) was inserted within intergenic region of the soybean chromosome, while T-DNA of a transgenic soybean line (10-19-1) located exon region of chromosome 13. This data of integration site and flanking sequences is useful for the biosafety assessment and for the identification of the β-carotene biofortified transgenic soybean.
A variety of genetically modified (GM) crops have been developed in Korea. In these crops, the resveratrol-enriched transgenic rice plant has moved ahead to generate the dossier for regulatory review process required for commercialization of GM crop. The resveratrol-enriched transgenic rice plant could be released to farmers for cultivation after national regulators have determined that it is safe for the environment and human health. Here we developed a PCR-based DNA marker based on flanking sequences of transgene for the discrimination of zygosity in resveratrol-enriched transgenic rice plant. This DNA marker will be useful for identifying of resveratrol-enriched transgenic rice plant, and can also be use to estimate transgene movement occurred by pollen transfer or seed distribution.