• Journal of Crop Science and Biotechnology
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• 제10권1호
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• pp.50-56
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• 2007

Barley S-adenosylmethionine synthetase1 gene, which was differentially expressed in seed development of extra early barley, was regulated by the phytohormones and abiotic stresses. In order to identify the regulation regions which were involved in transcriptional control of the phytohormones and abiotic stresses, we isolated 1459 bp fragment of HvSAMS1 gene promoter using genome walking strategy and deletion series were constructed. Deleted upstream fragments(-1459, -1223, -999, -766, -545, -301 bp) were fused to the GUS reporter gene and evaluated via Agrobacterium-mediated transient expression assay. Increased GUS activity of HvSMAS1 promoter -301/GUS construct under each of NaCl, $GA_3$, ABA and ethylene application was found. However, GUS activity was negligible in the leaves transformed with the HvSMAS1 promoter(-1459, -1223, -999, -766 and -545)/GUS constructs. No significant induction of GUS activity was observed for the ethionine and spermidine treatments. In order to locate promoter sequence of the HvSAMS1 gene that was critical for the activation of gene expression, deletion and addition promoter derivatives(+, includes 43 bp of 5' ORF) of the HvSAMS1 gene fused to the GUS reporter gene were applied. The tobacco leaves which harbored the additional HvSAMS1 promoter(-1459+, -1459 to -546, -545+ and -301+)/GUS construct did not significantly induce GUS activity as compared to the HvSAMS1 promoter(-1459, -545 and -301)/GUS constructs under each of NaCl, ABA and $GA_3$ treatment. However, the GUS activity was high in the tobacco leaves which harboring the -211 to -141 regions of the HvSAMS1 promoter. This result suggested that HvSAMS1 gene expression might be regulated by this region(from -211 to -141).

• 원예과학기술지
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• 제31권5호
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• pp.598-606
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• 2013

약은 생식과 화형을 결정짓는 꽃의 주요한 기관 중 하나이다. 오리엔탈나리인 아카풀코로부터 만든 약 특이적 cDNA library로부터 2000개의 ESTs를 무작위로 선발하였다. 잎과 약을 cDNA 탐침으로 이용한 differential slot blot이 약에서 발현되는 클론들을 얻기 위해 사용되었으며 570개의 비반복적 ESTs를 얻었고 염기서열분석을 하였다. BLASTX 알고리즘을 이용하여 GenBank에 비교해서 191개의 클론이 의미 있는 유사성을 보였지만 나머지(66.5%)는 기존에 보고된 염기서열에 확인되지 않았다. Gene ontology(GO) annotation에 따른 기능분류결과 대체적으로 세포와 세포구성 부분에서 주요하게 단백질이 확인되었다. 7개의 다른 기관과 발달 단계에서 전사체 분석은 약특이적일 적으로 추정되는 30개의 클론을 가지고 노던혼성화반응을 이용하여 수행하였다. 이러한 결과는 differential slot blot을 이용하여 약에 발현되는 유전자를 선별하는 것이 매우 효과적인 방법인 것으로 간주되며 또한 지금의 연구가 앞으로 나리의 화분을 포함한 약에 대한 기초정보를 제공할 수 있을 것으로 생각한다.

• Molecules and Cells
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• 제23권3호
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• pp.304-315
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• 2007

Fusarium graminearum causes a serious scab disease of small grains in Korea. The nucleotide sequence of the genomic RNA of a double-stranded RNA (dsRNA) virus, Fusarium graminearum virus-DK21 (FgV-DK21), from F. graminearum strain DK21, which is associated with hypovirulence in F. graminearum, was determined and compared to the genome sequences of other mycoviruses, including Cryponectria hypoviruses. The FgV-DK21 dsRNA consists of 6,624 nucleotides, excluding the 3'-terminal poly(A) tail. The viral genome has 53- and 46-nucleotide 5' and 3' untranslated regions (UTRs), respectively, and five putative open reading frames. A phylogenetic analysis of the deduced amino acid sequence of ORF1, which encodes a putative RNA-dependent RNA polymerase, and those of other mycoviruses revealed that this organism forms a distinct virus clade with other hypoviruses, and is more distantly related to other mycoviruses (3.8 to 24.0% identity). However, pairwise sequence comparisons of the nucleotide and deduced amino acid sequences of ORFs 2 through 5 revealed no close relationships to other protein sequences currently available in GenBank. Analyses of RNA accumulation by Northern blot and primer extension indicated that these putative gene products are expressed from at least two different subgenomic RNAs (sgRNAs), in contrast to the cases in other hypoviruses. This study suggests the existence of a new, as yet unassigned, genus of mycoviruses that exhibits a potex-like genome organization and sgRNA accumulation.

• Asian-Australasian Journal of Animal Sciences
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• 제19권5호
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• pp.622-626
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• 2006

Molecular characterization of Hallikar, the native cattle breed of Karnataka, was undertaken using 19 cattle specific, highly polymorphic microsatellite markers recommended by FAO. The genomic DNA was subjected to PCR amplification and alleles were resolved through six per cent denaturing PAGE with a 10 bp DNA ladder followed by silver staining. Genotyping of animals was done based on allele size. The number of alleles ranged from three to nine with allele sizes ranging from 102 bp to 294 bp. These alleles were distributed in the frequency range between 0.0306 and 0.8673 in the population. The mean observed number of alleles was $6.368{\pm}1.4225$. The mean observed and expected heterozygosities were $0.7515{\pm}0.1734$ and $0.7850{\pm}0.1381$, respectively. The high heterozygosity observed implies presence of higher genetic variability within Hallikar breed. The PIC (Polymorphism Information Content) values ranged from 0.2322 (ETH152) to 0.8654 (ETH225). The percentage of polymorphic loci obtained was 100 as all the 19 microsatellite markers were found to be polymorphic. Except for ETH152, all the other loci had high PIC values, indicating that these markers are highly informative for characterization of Hallikar breed. The population was tested for Hardy-Weinberg equilibrium at 19 microsatellite loci, and at 74 per cent of the loci the population was found to be in disequilibrium.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2022년도 추계학술대회
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• pp.292-292
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• 2022

Since 1992, the Rural Development Administration (RDA), Republic of Korea in collaboration with International Rice Research Institute (IRRI) has developed 6 japonica rice varieties(MS11, Japonica 1, 2, 6, 7 and Cordillera 4) that are adaptable to tropical regions. However, these varieties show moderate resistance or susceptibility to certain biotic and abiotic stress. The development of varieties with more stable forms of resistance is highly desirable, and this could be possibly achieved through rapid introgression of known biotic and abiotic resistant genes. In this study, we analyzed the allele types of major biotic stress resistant genes including Xa5, Xa13, Xa21 and Xa25 for bacterial leaf blight, Pi5, Pi40, Pish and Pita2 for blast, tsv1 for rice tungro spherical virus, and Bph6, Bph9, Bph17, Bph18 and Bph32 for brown planthopper by using gene-specific molecular markers. In addition, seed quality related genes Sdr4 for preharvest sprouting and qLG-9 for seed longevity were also analyzed. The results revealed that2h5 and Xa25 resistance alleles showed in all varieties while Pi5 resistance allele showed only in MS11. The Pish resistance allele were present in five varieties except for Japonica 1. Meanwhile, for the rest of the genes, no presence of resistance alleles found in six varieties. In conclusions, most of tropical japonica varieties are lack of the major biotic stress resistant genes and seed quality genes (Sdr4 and qLG-9). Moreover, the results indicated that rapid deployment of a few major genes in the current tropical japonica rice varieties is urgent to increase durability and spectrum of biotic stress resistance and also seed dormancy/longevity which are essential traits for tropical environments.

• Molecules and Cells
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• 제21권2호
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• pp.284-293
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• 2006

Even though Ac/Ds gene-tagging systems have been established in many higher plants, maize is the only major plant in which short-distance transposition of Ac/Ds has been utilized to probe gene function. This study was performed to evaluate the efficiency of obtaining new alleles and functional revertants from Ds insertion loci in rice. By analyzing 1,580 plants and the progeny of selected lines, the insertion sites and orientations of Ds elements within 16 new heritable alleles of three rice loci were identified and characterized. Intragenic transposition was detected in both directions from the original insertion sites. The closest interval was 35 bp. Three of the alleles had two Ds elements in cis configuration in the same transcription units. We also analyzed the excision footprints of intragenic and extragenic transpositions in Ds-inserted alleles at 5 loci. The 134 footprints obtained from different plants revealed predominant patterns. Ds excision at each locus left a predominant footprint at frequencies of 30-75%. Overall, 66% of the footprints were 7-bp additions. In addition, 16% of the excisions left 0-, 3-, 6-, and 9-bp additions with the potential of conserving reading frame.

• Molecules and Cells
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• 제25권2호
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• pp.205-210
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• 2008

To develop molecular markers linked to the $L^4$ locus conferring resistance to tobamovirus pathotypes in pepper plants, we performed AFLP with 512 primer combinations for susceptible (S pool) and resistant (R pool) DNA bulks against pathotype 1.2 of pepper mild mottle virus. Each bulk was made by pooling the DNA of five homozygous individuals from a T10 population, which was a near-isogenic $BC_4F_2$ generation for the $L^4$ locus. A total of 19 primer pairs produced scorable bands in the R pool. Further screening with these primer pairs was done on DNA bulks from T102, a $BC_{10}F_2$ derived from T10 by back crossing. Three AFLP markers were finally selected and designated L4-a, L4-b and L4-c. L4-a and L4-c each underwent one recombination event, whereas no recombination for L4-b was seen in 20 individuals of each DNA bulk. Linkage analysis of these markers in 112 $F_2$ T102 individuals showed that they were each within 2.5 cM of the $L^4$ locus. L4-b was successfully converted into a simple 340-bp SCAR marker, designated L4SC340, which mapped 1.8 cM from the $L^4$ locus in T102 and 0.9 cM in another $BC_{10}F_2$ population, T101. We believe that this newly characterized marker will improve selection of tobamovirus resistance in pepper plants by reducing breeding cost and time.

• 원예과학기술지
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• 제33권4호
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• pp.550-558
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• 2015

To analyze the linkage relationships among molecular markers recently reported to be linked to onion (Allium cepa L.) Ms, a restorer-of-fertility locus, in onion (Allium cepa L.), three single nucleotide polymorphism markers were converted into cleaved amplified polymorphic sequence (CAPS) markers based on onion transcriptome sequences and the rice genome database. Analysis of the recombinants selected from 4,273 segregating plants using CAPS and other linked markers demonstrated the jnurf13 and jnurf610 markers to perfectly co-segregate with the Ms locus. In contrast to jnurf13, the jnurf610 marker was not in perfect linkage disequilibrium with the Ms locus in diverse breeding lines. Thus, the jnurf13 marker and the marker for identification of cytoplasm types were utilized to enhance the efficiency of onion breeding through four applications. First, 89 maintainer lines containing the normal cytoplasm and homozygous recessive Ms genotypes were successfully identified from 100 breeding lines. Second, these two molecular markers were used to analyze the main sources of male-fertile contaminants frequently found in the male-sterile parental lines during F1 hybrid seed production. The majority of the contaminants contained heterozygous Ms genotypes, indicating that pollen grains harboring the dominant Ms genotype may have been introduced during propagation of the maintainer lines. Therefore, the genetic purity of the two maintainer lines was analyzed in the third application, and the results showed that both maintainer lines contained 13-21% off-types. Finally, the two markers were used to increase the seed yield potentials of two open-pollinated varieties containing sterile cytoplasms by removing the plants harboring homozygous recessive and heterozygous Ms genotypes.

• Journal of Plant Biotechnology
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• 제37권4호
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• pp.425-441
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• 2010

Forages are the backbone of sustainable agriculture. They includes a wide variety of plant species ranging from grasses, such as tall fescue and bermudagrass, to herbaceous legumes, such as alfalfa and white clover. Abiotic stresses, especially salinity, drought, temperature extremes, high photon irradiance, and levels of inorganic solutes, are the limiting factors in the growth and productivity of major cultivated forage crops. Given the great complexity of forage species and the associated difficulties encountered in traditional breeding methods, the potential from molecular breeding in improving forage crops has been recognized. Plant engineering strategies for abiotic stress tolerance largely rely on the gene expression for enzymes involved in pathways leading to the synthesis of functional and structural metabolites, proteins that confer stress tolerance, or proteins in signaling and regulatory pathways. Genetic engineering allows researchers to control timing, tissue-specificity, and expression level for optimal function of the introduced genes. Thus, the use of either a constitutive or stress-inducible promoter may be useful in certain cases. In this review, we summarize the recent progress made towards the development of transgenic forage plants with improved tolerance to abiotic stresses.

• Journal of Ginseng Research
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• 제41권4호
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• pp.444-449
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• 2017

The development of molecular markers is one of the most useful methods for molecular breeding and marker-based molecular associated selections. Even though there is less information on the reference genome, molecular markers are indispensable tools for determination of genetic variation and identification of species with high levels of accuracy and reproducibility. The demand for molecular approaches for marker-based breeding and genetic discriminations in Panax species has greatly increased in recent times and has been successfully applied for various purposes. However, owing to the existence of diverse molecular techniques and differences in their principles and applications, there should be careful consideration while selecting appropriate marker types. In this review, we outline the recent status of different molecular marker applications in ginseng research and industrial fields. In addition, we discuss the basic principles, requirements, and advantages and disadvantages of the most widely used molecular markers, including restriction fragment length polymorphism, random amplified polymorphic DNA, sequence tag sites, simple sequence repeats, and single nucleotide polymorphisms.