Ionic liquids (ILs) are organic salts with low melting point by asymmetric ionic strength between cation and anion. They have been known as promising DNA extraction, separation and preservation agent due to their hydrophilic, hydrophobic interaction with DNA. However, few studies have been performed about how DNA-ILs complexes form and their mechanism. Herein, we present three types of ionic liquids (EMIM-Cl, BMIM-Cl, and OMIM-Cl) change the DNA structure depend on alkyl chain length of ionic liquids. Structural changes of DNA by ionic liquids are observed by Atomic force microscopy, gel electrophoresis, zeta potential and solid-state nanopore technology. The results of AFM show the different structures of DNA, including aggregate, stretching, and bundling shapes in terms of EMIM-Cl, BMIM-Cl, and OMIM-Cl respectively. In DNA translocation experiment, DNA/EMIM-Cl show rare translocation signal due to aggregated structure by neutralized surface charge. DNA/BMIM-Cl and DNA/OMIM-Cl show slowing down the translocation speed due to changes of DNA net charge and structure. Especially, OMIM-Cl make slowing down the DNA translocation speed about 102~104 times compared to translocation speed of bare DNA by unzipping the bundling shape of complex. In conclusion, the morphology of DNA could be modified by the incorporation with different alkyl chain length of ILs, providing their further application in nanopore technique for slowing DNA sequencing or understanding protein-DNA interaction.
본 연구는 향후 송추이주사업으로 인한 수생태계 회복의 효과에 대한 검증을 위하여 사업 전 송추계곡의 상류부터 하류 구간을 대상으로 저서성 대형무척추동물의 정밀 모니터링을 실시하여 송추이주사업 이전의 수생태계 현황 자료를 구축하고자 한다. 현지조사는 2012년 11월부터 2013년 8월까지 총 3회에 걸쳐 조사를 실시하였으며, 조사구간은 대조구간(St. 1), 이주구간(St. 2, 3), 하류구간(St. 4)의 총 4개 지점을 선정하였다. 저서성 대형무척추동물은 총 4문 5강 9목 32과 62종 3,805개체가 출현하였다. 군집분석결과 대조구간에서 우점도 0.52(±0.16), 다양도 1.95(±0.44), 균등도 0.81(±0.04), 풍부도 2.25(±0.85)로 나타나 상대적으로 가장 안정된 군집구조를 유지하는 것으로 분석되었다. 출현 종수와 개체수의 EPT-비율은 대조구간에서 각각 67.60(±5.66)%, 66.30(±2.06)%로 가장 높게 나타났으며, 하류구 간으로 갈수록 감소하는 것으로 분석되었다. 기능군분석 결과 섭식기능군은 하류구간으로 갈수록 shredders의 비율은 감소하고 gathering-collectors와 filtering-collectors의 비율은 증가하였으며, 서식기능군은 하류구간으로 갈수록 sprawlers의 비율은 감소하고 burrows의 비율은 증가하는 것으로 나타났다. 하천오수생물지수(KSI)를 분석한 결과 대조구간은 평균 0.75(±0.10)로 A등급으로 나타났으며, 하류구간으로 갈수록 평균 지수값이 증가해 B등급으로 하락하 는 것으로 평가되었다.
무에 대한 I2 증기의 작물체 침적속도와 뿌리 전류계수를 측정하기 위하여 파종 후 29 일에서 53 일 사이에 생육시기별로 작물체를 I2 증기에 80 분 간 피폭시켰다. 피폭은 오전 중에 투명한 상자 내에서 수행되었다. 침적속도(ms-1)는 대체로 1.0×10-4∼2.0×10-4의 범위로 생육밀도가 높을수록 증가하는 경향이었다. 또한 상대습도가 높을 경우 값이 커진다는 기존 보고와 어느 정도 일치하였다. 본 침적속도는 몇몇 야외 측정치보다 수 십 배 정도 낮았고 이는 주로 피폭상자 내의 낮은 풍속(0.2 ms-1 내외)에 기인하는 것으로 추정되었다. 뿌리 전류계수(작물체 총침적량에 대한 수확시 뿌리 내 함유량의 비)는 다소 보수적으로 계산하여 파종 후 29 일 피폭에서 1.3×10-3, 파종 후 53 일 피폭에서는 5,0×10-3이었다. 본 실험결과의 이용에 있어서는 기상 조건, 요오드의 물리화학적 형태 등에 유의할 필요가 있다.
The relationship between source leaf position and photo-assimilate translocation and distribution was characterized for tomato (Lycopersicon esculentum Mill) grown in the greenhouse. Three different positions of source leaf on the stem (first node above or below the first fruit cluster and 5th node above the first fruit cluster) were tested for their influence on 14CO2 assimilation and transfer to different parts of the plant. The leaves at the 5th node above the first fruit cluster transferred the highest (57%) proportion of C14 to other plant parts, followed by leaves home on the first node below the first fruit cluster (50%), and the first node above the first fruit cluster (39%). In all treatments, fruits served as the strongest sink for C14, followed by stem, leaf, and root tissues. The leaf home on the 5th node above the first fruit cluster transferred the largest amount of C14 to the second fruit cluster.
This experiment was conducted to evaluate the effects of high temperature on the stem, leaf and grain of barley during the ripening period and to provide information for the development of high-temperature cultivation techniques and adaptive varieties. We used an artificial climate control facility, to provide a temperature 3℃ higher than the normal average temperature during the ripening stage. Although the maximum rate of starch synthesis was increased at high temperature by approximately 11%, the starch content was decreased, because the period of starch synthesis ended 4 days earlier. As in the case of starch synthesis, the expression of genes related to starch synthesis was increased at the early ripening stage in the high temperature treatment, however, the duration of expression tended to decrease rapidly. Furthermore, the partitioning rate of assimilation products in the panicle increased to a greater extent in the high temperature treatment than in the control. In contrast, for the stem and leaf, the partitioning rate of assimilation products decreased more rapidly in the high temperature treatment than in the control. On the basis of these results, it can be considered that the translocation rate of assimilation products increased to a greater extent in the high temperature treatment than in the control at the early ripening stage. These results indicate that the decrease in grain weight at high temperature during the ripening stage is attributable to an increase in the speed of starch synthesis at high temperature, but the increase in ripening speed does not compensate for the shortening of the ripening period. Finally to develop varieties and cultivation techniques suited to high temperature, we need to focus on physiological characteristics related to the duration of starch synthesis.
The wheat-rye translocations are world widely used in wheat breeding. Among the various forms of wheat-rye translocation, the 1RS (short arm of rye chromosome 1) translocations are the most widely used because of the valuable genes (e.g. biotic / abiotic resistance genes) introduced from rye. We have developed a new wheat-rye cultivar ‘TRANS’ which have freezing resistance and high yield in this study. ‘TRANS’ is a new wheat-rye translocation cultivar developed by crossing between common wheat ‘Keumkangmil’ and ‘951188-G3-G1’, a 1AL.1RS translocation derived from ‘Fleming’ with the aim of high yield and resistant genes to various unfavorable environments carried by 1RS. ‘TRNAS’ have clearly different genetic and agronomic traits to the control cultivar ‘Keumkangmil’. The heading date and maturity of ‘TRANS’ are later than that of ‘Keumkangmil’. ‘TRANS’ has 1023 spike number per square meters and grain yield of 541 kg/10a, which are higher than ‘Keumkangmil’ (904 spike number/m2 and 504 kg/10a yield). ‘TRANS’ showed winter hardiness and powdery mildew resistance in artificial infection test and field evaluation. ‘TRANS’ gives lower flour yield than ‘Keumkangmil’ but ash and protein content were similar to that of ‘Keumkangmil’. The color of flour and noodle dough of ‘TRANS’ were darker than ‘Keumkangmil’. Quality parameters related to milling, flour quality, noodle dough and end-use quality of ‘TRANS’ indicates that ‘TRANS’ is soft wheat suitable for noodle making. ‘TRANS’ can be cultivated in entire part of Korea. Registration Grant No.: 4695
Wheat-rye translocation lines are widely used in wheat breeding programmes by reason of biotic stress tolerances. Though there have been a number of researches regarding abiotic stress tolerance, the tolerance of the lines depends on wheat genetic background, not on rye chromosome. Here, we investigated wheat-rye translocation specific transcripts derived from cDNA-AFLP under drought stress, which may help to elucidate the reaction under the stress. ‘OK91G117’ (1BL.1RS translocation) and ‘OK91G144’ (non-translocation) were used as materials, which are near-isolines for 1RS. 25% PEG 6000 was added in culture solution to simulate drought condition and root tissues were sampled at each 0 h, 3 h, 6 h, 12 h, 24 h, and 48 h after PEG treatment for RNA extraction. As a result of cDNA-AFLP, TDFs (transcript derived fragments) that were specific to OK91G117 were sequenced. GO functions of each sequenced TDF were annotated by Blast2GO using standard parameter with cut-off level 3 and mapped to the GO term (i.e. biological process; BP, molecular function; MF, cellular component; CC). The term with “organic substance metabolic process”, “primary metabolic process”, and “cellular metabolic process” account for almost 50 % of BP. The most represented terms among probes classified to MF were “transferase activity” and most of TDF were annotated in “cell part” of CC. In addition, rye-chromatin specific markers were developed by BLAST comparing sequence of TDF with wheat and rye genome data. RT-PCR was conducted to validate expression patterns of selected TDF. Further studies will be needed to elucidate functions of the highly expressed genes under drought stress.
Common wheat has complex genome composition of homoeologous hexaploid (AABBDD, 2n = 6x = 42) and each homoeologous genome has high similarity. Due to these complexity, wheat genome study is a large challenge to researchers for genomic and genetic study. We analyzed expressions of individual wheat genome and rye genome specific transcripts using custom array with 2BS.2RL wheat-rye translcoation. Genomic probes were synthesized within each diploid progenitors (AA, BB, DD, 2n = 14, respectively) of wheat, common wheat, and rye (RR, 2n = 14). Total RNA isolated from seedlings of T. urartu, Ae. speltoides, Ae. squarrosa, ‘Chinese Spring’, ‘Chaupon’, and 2BS.2RL were hybridized on arrays. Each homoeologous gene differentially expressed in hexaploid wheat and rye were identified on the custom array and the transcripts were clustered based on hybridization values. qRT-PCR was performed to verify the custom array result with a set of five genes by highly replicated experiments (three biological and three technical replications). The qRT-PCR results demonstrated genome specific expression of five genes in sympathy with array results. Here we provide information of each individual genome specific transcripts and it will we a useful data to study complex wheat genome compositions.
Wheat-rye translocation lines were developed to produce a main crop resistant to biological and physical stress. 'Chaupon' rye contains 2RL chromatin to harbor resistance genes for powdery mildew and leaf rust. In order to identify chromosome 2RL-derived rye proteins and 2RL-perturbed proteins in wheat-rye translocation lines, the gel-based proteomics was employed with 'Coker797' (non-2RL), 'Hamlet' (2RL) and 'near-isogenic line' (stabilized 2RL). The leaf proteome was resolved on 2D-gel, resulting in 216 spots in a final selection. A total of 90 proteins were identified with the identification success rate of 42%. The identified proteins were classified by functional annotation: metabolism (64%), cellular process (5%), translation (2%), regulatory function (1%) and hypothetical (28%). The proteins belonged to metabolism were subdivided into carbohydrate metabolism (36%), energy metabolism (35%), metabolism of lipid, amino acid, other amino acid and biosynthesis of secondary metabolites (each 6%) and others (5%). A total of 53 proteins were differentially expressed, in which β-glucosidase, in particular, originated from the chromosome 2RL of rye, was exclusively appeared in NIL. In addition, small Ras-related GTP binding-protein assigned to wheat was predominantly found in 2RL rye chromatin-possessing NIL. These results suggest that the acquired genetic traits obtained from rye 2RL enhance the resistance to biotic and abiotic stress in wheat-rye translocation lines by altered the proteome expression. In leaf metabolome analysis, 11 predominant metabolites containing trans-aconitate, glutamate, and betaine were identified by 1H-NMR-based metabolite fingerprinting. The overall metabolites pattern of NIH appears to be closer to Coker797 rather than Hamlet. Thus, the metabolic phenotype of NIL was not so much lineated from Hamlet contrast to proteomic phenotyping.
Rye (Secale cereale L.) chromatins have been used to introduce agronomically important traits into wheat (Triticum aestivum L.). Wheat-rye translocations in the form of 1RS.1AL, 1RS.1BL, 2BS.2RL have been developed for an important genetic source of disease and pest resistance. The long arm of rye chromosome 2 (2RL) has valuable genes that confer resistance to pests such as biotype L of Hessian fly, powdery mildew, leaf and stem rust. Here, we report the generation and analysis of expressed sequence tags from Hessian fly infested wheat-rye translocation. RNAs were isolated from young seedlings infested by Hessian fly. cDNA library was constructed using Clontech cDNA library construction kit. Random sequencing of candidate clones were performed. The EST clones might be useful to clone target gene sequences and would provide clues on molecular interaction between wheat and Hessian fly.
The wheat-rye translocation lines have been agriculturally developed for the resistance to the biotypes of Hessian fly as a major insect pest of wheat. In order to compare the proteomic profiles between ‘Coker797’ (non-2RL), ‘Hamlet’ (2RL), and near-isogenic line (NIL) carrying 2RL, we evaluated the protein extraction and preparation methods for two-dimensional gel electrophoresis approach. The tissues such as leaves, stems, and roots from three wheat-rye lines were extracted by following trichloroacetic acid (TCA)/acetone precipitation. In a preliminary proteome analysis, a commonly expressed protein in Hamlet and NIL strain was identified as methionine synthase annotated in Hordeum vulgare subsp. The present study will provide the experimental guideline for the proteomic study of other useful crop plant tissues.