본 연구는 옥수수 재배 시 환경에 영향을 미치는 노균병 저항성과 관련된 유전자 후보군을 탐색해서 노균병으로 인한 토양오염과 옥수수 생산량 감소를 해결하기 위하여 노균병 저항성 품종을 효율적으로 발굴하기 위한 연구이다. 옥수수의 6번 염색체의 152,892,333과 154,335,437 사이에 있는 노균병 저항성 유전자를 탐색하였으며 이 부분에 존재할 것으로 예상되는 전사체에서 38개의 프라이머 세트를 디자인하여 이 중 16개의 예측 전사체를 가려 내었다. 또한 RT-PCR을 수행하여 감염된 Ki11의 발현이 높은 7개의 전사체로 5개의 품종에 대하여 건강한 샘플과 감염된 샘플을 검정하였고 최종 5개의 후보 유전자군[알려지지 않은 미확인 유전자 2개, OFP transcription factor, bZIP transcription factor, pentatricopeptide repeat (Ppr)]이 발견 되었다. 본 연구의 결과로 추가적인 실험 설계를 통해 5개의 후보 유전자군에 대한 재검정을 통하여 확실한 노균병 저항성 유전자를 발굴하고 이를 노균병 저항성 품종 개발 및 방재에 이용할 수 있을 것으로 사료된다.
Drought is one of important environmental stress for plant. Drought has deleterious effect to plant growth including maize (Zea mays L.) such as vegetative and/or reproductive growth, root extension, photosynthesis efficiency, flowering, anthesis-silking interval (ASI), fertilization, and grain filling. In this study, we screened drought tolerant maize in 21 cultivars from different sources, sixteen NAM parent lines (B73, CML103, CML228, CML247, CML277, CML322, CML333, CML69, Ki11, Ki3, Ky21, M37W, Mo18w, NC350, Oh43 and Tx303), four Korean hybrids (Cheongdaok, Gangdaok, Kwangpyeongok and Pyeonganok) and one Southeast Asian genotype (DK9955). Drought stress (DS) index was evaluated with leaf rolling score at seedling stage and ASI at silking date. The leaf rolling scoring of CML228, DK9955 and Ki11 were determined 1.28, 1.85, 1.86, respectively. However, M37W, Kwangpyeongok, B73 and NC350 were determined over the 3. ASI analysis revealed that CML228, CML103, Cheongdaok, NC350, B73, CML322, Kwangpyeongok and Ki11 are represented less than 5 days under DS and less than 3 days of difference between DS and well-watered (WW), but CML69, Ki3, Pyeonganok, M37W, Mo18w and Gangdaok were represented more than 10 days under DS and more than 8 days of difference between DS and WW. Multi-Dimensional Scaling (MDS) analysis determined CML228, Ki11, and CML322 were regarded as drought tolerance cultivars. Eventually, Ki11 showed genetic similarity with Korean cultivars by QTL analysis and MDS analysis. Ki11 has a potential for development of drought tolerance maize with Korean cultivars.
We screened the drought tolerant maize using seventeen maize genotypes from different sources, nine inbred genotypes from United States Department of Agriculture (USDA) (B73, CML103, CML228, CML277, CML322, CML69, Ki3, Ki11, and NC350), three Southeast Asian genotypes (DK9955, LVN-4, and 333), and five Korean hybrids (Cheongdaok, Gangdaok, Ilmichal, Kwangpyeongok, and Pyeonganok). We evaluated anthesis-silking interval (ASI), leaf senescence (LS), ears per plant (EPP), tassel length (TL), and fresh weight (FW) at silking date. According to ASI and LS examination, CML103 and Kill were drought tolerant genotypes, wheareas Ki3 and 333 were drought susceptible. EPP, TL, and FW differed according to drought resistance. Grain yield was correlated strongly with ASI, but moderately with LS. Difference in ASI between drought-stressed (DS) and well-watered (WW) conditions was less than three days in CML228, CML103, Cheongdaok, NC350, B73, Ki11, CML322, and Kwangpyeongok, whereas that of Ki3, Pyeonganok, and Gangdaok was more than 6.5 days. We concluded that CML228, CML103, Cheongdaok, NC350, B73, Ki11, CML322, and Kwangpyeongok are drought tolerant genotypes, whereas Ki3, Pyeonganok, and Gangdaok are drought susceptible.
This study was conducted to evaluate maize downy mildew resistance using spreader row technique in Cambodia. A total of forty maize lines were used in this experiment. Seven Korean varieties and seven breeding lines showed high infection rates (80~100%) and highly susceptible (HS) to downy mildew disease in both spring and fall. Also most of nested association mapping (NAM) parent lines were highly susceptible (HS). Meanwhile three inbred lines, Ki3, Ki11, and CML228, showed highly resistant (HR) or resistant (R) in spring and moderately resistant (MR) in fall. These three lines were already known as resistant inbred lines against downy mildew disease. It appears that spreader row technique was suitable for selection of maize downy mildew resistance in Cambodia. The incidence of downy mildew was influenced by weather conditions, especially relative humidity and temperature. Among several inoculation methods to screen for downy mildew resistance, this spreader row technique is effectively and easily used in the field of Southeast Asia.
Drought stress has detrimental effects on the seedling development, vegetative/ reproductive growth, photosynthesis, root proliferation, anthesis, anthesis-silking interval (ASI), pollination and grain yield in maize. Typically, two weeks before silking through pollination are an important time in maize life. Here we reviewed the effects of drought stress on growth, physiological/ molecular researches for drought tolerance, and breeding to genomics in maize. Drought stress during kernel development increases leaf dying and lodging, decreases grain filling period and grain yield. Physiological factors of drought stress/ effects are water content, water deficits, and water potential. Nowdays molecular marker assisted breeding method is becoming increasingly useful in the improvement of new germplasm with drought stress tolerance.
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 this study, we report that the development of a multiplex PCR method using species-specific primers for the simultaneous detection of Poaceae family members, including adlay, barley, maize, rice and wheat, based on the sequence polymorphism of the DNA-directed RNA polymerase beta'' chain (rpoC2) genes Species-specific primers were constructed with common forward primer and each reverse primers which have differences on the basis of sequences. Each primer pairs could amplify PCR products of 443 bp for rice, 346 bp for barley, 278 bp for adlay, 221 bp for wheat and 96 for maize, respectively, from the five chloroplast DNAs. The series of template DNA concentrations were identified by the sensitivity of multiplex PCR. The band of products were clearly amplified from the DNA concentration range of 0.01 to 10 ng/μL. In addition, the species-specific primers were examined for the detection of seven commercial flour mixed products. The combination of the sensitivity of a multiplex PCR with the specificity of the primers for the detection of species would allow to be applied in analyses of processed foods.
The objective of this study was to evaluate the drought tolerance in maize seedling using leaf rolling. Nineteen maize resources, seven Nested Association Mapping parents lines, six Korean commercial cultivars, and six Southeast Asia commercial cultivars, were used to examine drought tolerance. The leaf rolling scores were measured on each leaf in three stress conditions with moderate drought (10%), severe drought (7%), and extreme drought (5%). Generally leaf rolling score of seedlings increased at the lower soil water potentials (5~7%). As a result, drought-tolerant cultivars showed lower leaf rolling score (below 2.5) than the drought sensitive cultivars (above 3.5). Nine varieties, NK4043, CML322, DK9955, NK4300, Ki11, DK8868, CML228, LVN99, and LVN10, have been selected for tolerance to drought stress. These results suggest that the leaf rolling score in maize seedling has been made available to indirect index for drought tolerance.
Transcription factors are essential for the regulation of gene expression in plant. They are binding to either enhancer or promoter region of DNA adjacent to the gene and are related to basal transcription regulation, differential enhancement of transcription, development, response to intercellular signals or environment, and cell cycle control. The mechanism in controlling gene expression of transcription can be understood through the assessment of the complete sequence for the maize genome. It is possible that the maize genome encodes 4,000 or more transcription factors because it has undergone whole duplication in the past. Previously, several transcription factors of maize have been characterized. In this review article, the transcription factors were selected using Pfam database, including many family members in comparison with other family and listed as follows: ABI3 / VP1, AP2/EREBP, ARF, ARID, AS2, Aux/IAA, BES1, bHLH, bZIP, C2C2-CO-like, C2C2-Dof, C2C2-GATA, C2C2- YABBY, C2H2, E2F/DP, FHA, GARP-ARR-B, GeBP, GRAS, HMG, HSF, MADS, MYB, MYB-related, NAC, PHD, and WRKY family. For analyzing motifs, each amino acid sequence has been aligned with ClustalW and the conserved sequence was shown by sequence logo. This review article will contribute to further study of molecular biological analysis and breeding using the transcription factor of maize as a strategy for selecting target gene.
Recently, the importance of food safety is increasing due to numerous junk food. Junk food means to violate the law in stage such as production, manufacture, distribution, and sale of food. Many crop plants are processing as foods including bread, noodle, and other foods for supporting nutrition to human. For example, rice is one of the most well-known food crops in the world, and processed rice is being mixed with other processed crops to health food. The object of this study is to detect amount of specific grains, i.e. rice from processed foods mixed with other cereals. This experiment was performed to the following two steps: 1) designed the specific primer sets based on chloroplast DNAs, 2) amplified products using real-time PCR. We designed eleven primer sets within chloroplast DNA of rice, and then the confirmation of primer efficiency was to amplified with rice genomic DNA using real-time PCR. In addition, these primer sets were applied in other crops such as wheat, maize, and adlay to confirm specificity to rice. The rice specific primer sets were determined by the number of amplification and the melting peak through real-time PCR. As a result, five primer sets were confirmed to uniqueness in the rice genome. In conclusion, the specific primer sets would be useful for identifying rice grain from the processed foods to eliminate junk foods and for contribution of food safety.
Maize is one of the most important food and feed crops in the world including Southeast Asia. In spite of numberous efforts with conventional breeding, the maize productions remain low and the loss of yields by drought and downy mildew are still severe in Asia. Genetic improvement of maize has been performed with molecular marker and genetic engineering. Because maize is one of the most widely studied crop for its own genome and has tremendous diversity and variant, maize is considered as a forefront crop in development and estimation of molecular markers for agricultural useful trait in genetics and breeding. Using QTL (Quantitative Trait Loci) and MAS (Marker Assisted Breeding), molecular breeders are able to accelerate the development of drought tolerance or downy mildew resistance maize genotype. The present paper overviews QTL/MAS approaches towards improvement of maize production against drought and downy mildew. We also discuss here the trends and importance of molecular marker and mapping population in maize breeding.
We used a microarray dataset that is deposited in the public database to evaluate plant responses to heat stress and selected two genes, OsSHSP1 (Os03g16030) and OsSHSP2 (Os01g04380), that are highly expressed under heat stress in rice. OsSHSP1 and OsSHSP2 gene transcripts were highly induced in response to salt and drought. In addition, OsSHSP1 and OsSHSP2 gene transcripts were induced under ABA and SA. Subcellular localization of proteins of 35S::OsSHSP1 were associated with the cytosol, whereas those of and 35S::OsSHSP2 were associated with the cytosol and nucleus. Heterogeneous overexpression of both genes exhibited higher germination rates than those of wild-type plants under the salt treatment, but not under heat or drought stress. The network of both genes harboring 9 sHSPs as well as at least 13 other chaperone genes might support the idea of a role for sHSPs in the chaperone network. Our findings might provide clues to shed light on the molecular functions of OsSHSP1 and OsSHSP2 in response to abiotic stresses, especially heat stress.
Trypsin inhibitors occur in a wide range of foods, they have been studied extensively in plants such as soybean (Glycinemax) as the majority of species are considered as important food sources. Selected soybeans were subjected to various treatments including microwave heating, supercritical fluid CO2 de-fatting, ultrasound incubating by different treat time (5min-120min) to determine the effect on soybean trypsin inhibitor (STI) activities compared to the non-treatment. The results indicated that microwave reduced trypsin inhibitors ranged from 20.57±1.40 TIU/mg to 14.60±1.48 TIU/mg compared to the control (21.44±1.04 TIU/mg) respectively (p<0.05). The inactivation of STI activities by ultrasound ranged from 28.47±0.16 TIU/mg to 19.86±0.78 TIU/mg compared to the control (28.87±0.18 TIU/mg) respectively (p<0.05). While the observed STI activities by supercritical CO2 de-fatting was not significant compared to the control. This study signifies the effective potential of inactivation of trypsin inhibitor by several applicable treatments which might be benefit in the food manufacture and livestock feeding industry.
To develop the STS (Sequence Tagged Site) marker for discrimination between oat cultivars used the EEG (Euchromatin Enriched Genomic) DNA library in oat. The EEG DNA library was constructed the Mcr A and Mcr BC system in DH5 alpha bacterial cell line. About 3,500 EEG colonies had been constructed by using junk DNA exclusion. About 800 colonies were selected that included insert DNA more than 500 bp. It was analyzed the genetic information by using blast searches of NCBI web site. More than three hundred STS primer sets were developed using sequencing data of selected colonies and about 90 primer sets which showed single band were selected in Olgwiri. It was applied to twelve oat cultivars including Olgwiri and has been shown polymorphism at 15%. PCR product which amplified with selected STS primer was treated with six endonucleases and was showed polymorphic bands. These primers could be useful for specific allele tagging in mapping populations and germplasm and for the study of functional genomics of oat.