Specific Protein Spots Research Articles

Development of Thermotolerant Lactobacilli Cultures with Improved Probiotic Properties Using Adaptive Laboratory Evolution Method.

Probiotics play a significant role in functional foods. Heat stress and dehydration are the two principal mechanisms leading to inactivation and loss of probiotics viability in its production. There is a need to develop an industrial organism to withstand higher temperatures during its processing and storage. This current study aims to develop thermotolerant strains ofLacticaseibacillus casei N (N) andLactobacillus helveticusNRRL B-4526 (H) by acclimatizing the wild-type strains to the higher temperature of 45°C by adaptive laboratory evolution. A two-fold increase in biomass was observed in both acclimatized strains up to the 200th generation, which subsequently remained stable after 500 generations. The morphological change of these acclimatized strains was observed under scanning electron microscopy. Also, there was an increase in probiotic attributes of these acclimatized strains compared to their wild-types. Among two acclimatized strains, L. casei N-45 had shown higher tolerance in the acidic pH 3.0 (89.31%), the bile of 0.3% (84.45%), simulated gastric juice (79.12%), and simulated intestinal juice (73.86%). There was also an increase in salt tolerance (NaCl), radical scavenging activity, autoaggregation, coaggregation, and hydrophobicity of these adapted strains. The total protein profiling using 2D gel electrophoresis reveals the differences in protein expressions between wild-type and acclimatized strains. Specific protein spots from acclimatized strains of H-45 and N-45 were further subjected to MALDI-TOF MS/MS. Some of the identified proteins were recognized to play a role in RNA chaperones and protein synthesis during stress conditions.

Phosphorylation/dephosphorylation response to light stimuli of Symbiodinium proteins: specific light-induced dephosphorylation of an HSP-like 75 kDa protein from S. microadriaticum.

BackgroundSome genera of the family Symbiodiniaceae establish mutualistic endosymbioses with various marine invertebrates, with coral being the most important ecologically. Little is known about the biochemical communication of this association and the perception and translation of signals from the environment in the symbiont. However, specific phosphorylation/dephosphorylation processes are fundamental for the transmission of external signals to activate physiological responses. In this work, we searched phosphorylatable proteins in amino acids of Ser, Thr and Tyr from three species of the family Symbiodiniaceae, Symbiodinium kawagutii, Symbiodinium sp. Mf11 and Symbiodinium microadriaticum.MethodsWe used specific antibodies to the phosphorylated aminoacids pSer, pThr and pTyr to identify proteins harboring them in total extracts from three species of Symbiodinium in culture. Extractions were carried out on logarithmic phase growing cultures under a 12 h light/dark photoperiod. Various light/dark, nutritional and other stimuli were applied to the cultures prior to the extractions, and proteins were subjected to SDS-PAGE and western immunoblotting. Partial peptide sequencing was carried out by MALDI-TOF on specific protein spots separated by 2D electrophoresis.ResultsAt 4 h of the light cycle, several Thr-phosphorylated proteins were consistently detected in the three species suggesting a genus-dependent expression; however, most Ser- and Tyr-phosphorylated proteins were species-specific. Analysis of protein extracts of S. microadriaticum cultures demonstrated that the level of phosphorylation of two Thr-phosphorylated proteins with molecular weights of 43 and 75 kDa, responded inversely to a light stimulus. The 43 kDa protein, originally weakly Thr-phosphorylated when the cells were previously adapted to their 12 h dark cycle, underwent an increase in Thr phosphorylation when stimulated for 30 min with light. On the other hand, the 75 kDa protein, which was significantly Thr-phosphorylated in the dark, underwent dephosphorylation in Thr after 30 min of the light stimulus. The phosphorylation response of the 43 kDa protein only occurred in S. microadriaticum, whereas the dephosphorylation of the 75 kDa protein occurred in the three species studied suggesting a general response. The 75 kDa protein was separated on 2D gels as two isoforms and the sequenced spots corresponded to a BiP-like protein of the HSP70 protein family. The presence of differential phosphorylations on these proteins after a light stimulus imply important light-regulated physiological processes in these organisms.

Proteomic analysis reveals the differential proteins of endosperm between soft and hard wheat varieties

Abstract In this study, we revealed the differential proteins from the wheat endosperms using proteomic analysis and investigated their surface properties. The pattern of the polypeptides obtained from the Yangmai-15 and Yangmai-16 wheat varieties were compared using two-dimensional polyacrylamide gels. In addition, we compared the characteristics of the grain such as grain hardness, protein content, wet gluten, dough development time, dough stability, gliadin and glutenin contents between Yangmai-15 and Yangmai-16, and the results were significantly different. Notably, 216 and 197 protein spots were separated from Yangmai-15 and Yangmai-16, respectively. The isoelectric points of the identified proteins ranged from 4 to 10 and the molecular weights of proteins varied from 10 to 100 kDa. Further, 21 and 8 specific differential protein spots were identified from the flour of Yangmai-15 and Yangmai-16, respectively. The surface properties of identified peptides consisted of hydrophobic or hydrophilic residues, as well as randomly scattered residues. The proteomic analysis of the wheat endosperms provides a novel insight into the biochemical basis for the differences in physicochemical properties between the soft and hard wheat varieties.

Two-Dimensional Gel Electrophoresis Seed Proteome Map of Pigmented and Non Pigmented Sorghum Genotypes (Sorghum bicolor L. Moench)

Two-dimensional gel electrophoresis has been extensively used for separation and characterization of proteins. A protocol was established for two-dimensional gel electrophoresis (2-DE) seed proteome map of six sorghum genotypes including non-pigmented lines IS 3477, IS 33095, IS 7005 and pigmented lines are 2898, IS 7155 and IS 1202 in the pH range of 3-10. Proteins extracted from flour in a TCA/Acetone buffer were focused after cup-loading onto IPG strips. Successful separation in the second dimension was achieved using gradient gels in a horizontal SDS-PAGE system. The 2-D gels of six genotypes stained with Coomassie Brilliant Blue (CBB) which were resolved a total of 116, 140, 109, 98, 80 and 102 protein spots in IS 3477, IS 33095, IS 7005, IS 2898, IS 7155 and IS 1202 respectively. Furthermore, we have identified different protein spots by using PD-Quest software among the genotypes which were included Up-regulation protein spots (162), Downregulation protein spots (483), Qualitative protein spots (91), Quantitative protein spots (109), Common protein spots (542) and Specific protein spots (93). The presented 2- D patterns and identifications will be used to describe proteome differences between cultivars.

2D-DIGE Analysis of Eye Lens Proteins as a Measure of Cataract Formation.

This chapter describes the basics of two-dimensional difference gel electrophoresis (2D-DIGE) for multiplex analysis of two distinct proteomes. The example given describes the analysis of male and female rat lens soluble proteins labeled with fluorescent Cy3 and Cy5 dyes in comparison to a pooled standard labeled with Cy2. After labeling the proteomes are mixed together and electrophoresed on the same 2D gels. Scanning the gels at wavelengths specific for each dye allows direct overlay the two different proteomes. Differences in abundance of specific protein spots can be determined through comparison to the pooled standard.

A Two-Dimensional Difference Gel Electrophoresis (2D-DIGE) Protocol for Studies of Neural Precursor Cells.

This chapter describes the basics of two-dimensional difference gel electrophoresis (2D-DIGE) for multiplex analysis of up to distinct proteomes. The example given describes the analysis of undifferentiated and differentiated neural precursor cells labelled with fluorescent Cy3 and Cy5 dyes in comparison to a pooled standard labelled with Cy2. After labelling, the proteomes are mixed together and electrophoresed on the same 2D gels. Scanning the gels at wavelengths specific for each dye allows direct overlay of the two different proteomes and the differences in abundance of specific protein spots can be determined through comparison to the pooled standard.

Simultaneous Two-Dimensional Difference Gel Electrophoresis (2D-DIGE) Analysis of Two Distinct Proteomes.

This chapter describes the basics, applications, and limitations of two-dimensional gel electrophoresis (2DE) and two-dimensional difference gel electrophoresis (2D-DIGE) for multiplex analysis of distinct proteomes. We also propose a basic protocol for 2D-DIGE, technique that allows the analysis of paired protein extracts, which are labeled with fluorescent Cy3 and Cy5 dyes and electrophoresed with a Cy2-labeled standard extract on the same 2DE gels. Scanning the gels at wavelengths specific for each dye allows direct overlay the two different proteomes and the differences in abundance of specific protein spots can be determined.

Selection for higher fertility reflects in the seminal fluid proteome of modern domestic chicken

The high egg-laying capacity of the modern domestic chicken (i.e. White Leghorn, WL) has arisen from the low egg-laying ancestor Red Junglefowl (RJF) via continuous trait selection and breeding. To investigate whether this long-term selection impacted the seminal fluid (SF)-proteome, 2DE electrophoresis-based proteomic analyses and immunoassays were conducted to map SF-proteins/cytokines in RJF, WL and a 9th generation Advanced Intercross Line (AIL) of RJF/WL-L13, including individual SF (n=4, from each RJF, WL and AIL groups) and pools of the SF from 15 males of each group, analyzed by 2DE to determine their degree of intra-group (AIL, WL, and RJF) variability using Principal Component Analysis (PCA); respectively an inter-breed comparative analysis of intergroup fold change of specific SF protein spots intensity between breeds. The PCA clearly highlighted a clear intra-group similarity among individual roosters as well as a clear inter-group variability (e.g. between RJF, WL and AIL) validating the use of pools to minimize confounding individual variation. Protein expression varied considerably for processes related to sperm motility, nutrition, transport and survival in the female, including signaling towards immunomodulation. The major conserved SF-proteins were serum albumin and ovotransferrin. Aspartate aminotransferase, annexin A5, arginosuccinate synthase, glutathione S-transferase 2 and l-lactate dehydrogenase-A were RJF-specific. Glyceraldehyde-3-phosphate dehydrogenase appeared specific to the WL-SF while angiotensin-converting enzyme, γ-enolase, coagulation factor IX, fibrinogen α-chain, hemoglobin subunit α-D, lysozyme C, phosphoglycerate kinase, Src-substrate protein p85, tubulins and thioredoxin were AIL-specific. The RJF-SF contained fewer immune system process proteins and lower amounts of the anti-inflammatory/immunomodulatory TGF-β2 compared to WL and AIL, which had low levels- or lacked pro-inflammatory CXCL10 compared to RJF. The seminal fluid proteome differs between ancestor and modern chicken, with a clear enrichment of proteins and peptides related to immune-modulation for sperm survival in the female and fertility.

Proteomic analysis reveals the fungal resistance of soft wheat during storage

The prevention of fungal growth is of great importance for the safe storage of wheat. Previous studies have shown that soft wheat is more resistant to the xerophilic fungus Aspergillus glaucus than hard wheat during storage under the same conditions. We conducted a comparative proteomic analysis of embryos of soft wheat cv. Yangmai 15 and of hard wheat cv. Yangmai 16 to identify specific differentially expressed proteins. Dot-blot analysis of embryo lysates revealed 65 and 15 specific differential protein spots from Yangmai 15 and Yangmai 16, respectively. Four specific differential protein spots were revealed from Yangmai matched antifungal proteins, including a peroxidase, endogenous alpha-amylase/subtilisin inhibitor, xylanase inhibitor protein 1, and puroindoline B, while spots from Yangmai 16 did not match any antifungal proteins. This proteomic study provided novel insights to investigate the discrepancy of storage tolerances among different wheat varieties.

Analysis of Differential Proteins in Two Wing-Type Females of Sogatella furcifera (Hemiptera: Delphacidae).

Sogatella furcifera (Horvath) is an important rice pest with the wing dimorphism, including macropterous and brachypterous morphs. The protein expression profiles in two wing-type adults and two wing-type disc fifth-instar nymphs were analyzed using two-dimensional gel protein electrophoresis and mass spectrometry. In adults and fifth-instar nymphs, 127 and 162 protein spots were detected, respectively. Fifty-five differentially expressed protein spots were identified between the long-winged adults and the short-winged adults, and 62 differentially expressed protein spots were found between the long-winged disc fifth-instar nymphs and short-winged disc fifth-instar nymphs. In long-winged and short-winged adults, six and seven specific protein spots were identified, respectively, with five and seven protein spots having more than threefold increased level, respectively. In long-winged and short-winged disc morph nymphs, 8 and 12 specific protein spots were identified, respectively, with 11 and 17 spots containing more than threefold increased level, respectively. Among the 16 identified proteins, five proteins are associated with muscle function, suggesting that muscle is a main tissue where the genes were differentially expressed between the two wing types. In addition, the content of a peptidase with an insulinase domain was higher (by 3.02 ± 0.59 fold) in the short-winged fifth-instar nymphs than in the long-winged fifth-instar nymphs, which suggests that this peptidase may be involved in wing differentiation by regulating insulin receptors. The results of this study provide some genetic clues for the wing differential development in S. furcifera and provide more references for future studies.

Proteomics Study of Wheat (Triticum durumL.) Genotypes in Response to Drought Stress

Under present investigation two-dimensional electrophoresis (2-DE) technique was used for finding out the drought responsiveness of differentially expressed protein spots (DEPs) in the three drought susceptible (HI-8498, GW-1139 and MPO-1215) and three drought tolerant wheat genotypes (GW-1, AR-07-30 and AR-07-33). Artificial drought was induced by 20% (−0.52 bar) polyethylene glycol (PEG-6000) treatment. Total 630 protein spots were observed amongst all the genotypes which include 272 droughts responsive protein spots (DRPs), 37 differentially expressed protein spots (DEPs) and 76 cultivar specific protein spots. Susceptible genotypes, namely, HI-8498, GW-1139 and MPO-1215 showed 59.18, 51.37 and 48.71% of DRPs respectively whereas the tolerant genotypes viz. GW-1, AR-07-30 and AR-07-33 expressed 50.89, 25.38 and 40.83% DRPs respectively. Among all the genotypes examined, the susceptible genotypes (HI-8498 and GW-1139) and tolerant genotype (GW-1) showed better performance under the stress conditions. Using this technique the drought responsive proteins were identified in both susceptible and tolerant wheat genotypes which can prove to be an important tool for understanding the biochemical mechanism in the plants under the stress conditions.

Abundant storage protein depletion from tuber proteins using ethanol precipitation method: Suitability to proteomics study.

High-abundance proteins (HAPs) hamper in-depth proteome study necessitating development of a HAPs depletion method. Here, we report a novel ethanol precipitation method (EPM) for HAPs depletion from total tuber proteins. Ethanol showed a dose-dependent effect on depletion of sporamin from sweet potato and patatin from potato tubers, respectively. The 50% ethanol was an optimal concentration. 2DE analysis of EPM-prepared sweet potato proteins also revealed enrichment of storage proteins (SPs) in ethanol supernatant (ES) resulting in detection of new low-abundance proteins in ethanol pellet (EP), compared to total fraction. The ES fraction showed even higher trypsin inhibitor activity than total proteins, further showing the efficacy of EPM in enrichment of sporamin in ES fraction. Application of this method was demonstrated for comparative proteomics of two sweet potato cultivars (Hwang-geum and Ho-bac) and purification of SP (sporamin) in its native form, as examples. Comparative proteomics identified many cultivar specific protein spots and selected spots were confidently assigned for their protein identity using MALDI-TOF-TOF analysis. Overall, the EPM is simple, reproducible, and economical for depletion of SPs and is suitable for downstream proteomics study. This study opens a door for its potential application to other tuber crops or fruits rich in carbohydrates.

NLRs Challenge Impacts Tight Junction Claudins In Sertoli Cells.

The present study aims to investigate the NALP3 system and its effect on claudins in Sertoli cells using a mouse adult Sertoli cell line as a model. We focus on the Sertoli cell biology looking for the possible implications for male reproductive functions. Adult Sertoli cells were transfected with NAPL3 siRNA and treated with NOD1 (ie-DAP) and NOD2 (MDP) receptor ligands. Two dimensional gel electrophoresis was performed on lysates of non-challenged and MDP-treated Sertoli cells. There were positive claudin-5 and claudin-11 expression levels on transcript (RT-qPCR) levels. Specific protein spots in 2D gels were detected after bioinformatics analysis. This study demonstrates direct induction of tight-junction proteins probably favouring junction stability. The innate immunity and tight-junction pathway integration probably have a protective role for both blood-testis immune barrier and spermatogenesis compartmentalisation maintained by the very same barrier. This integration also points the way for mechanistic research of the disturbances inflicted during an inflammatory response in the testis niche.

Differentiation between fresh and frozen–thawed sea bass (Dicentrarchus labrax) fillets using two-dimensional gel electrophoresis

This study aimed to identify a protein marker that can differentiate between fresh skinless and frozen–thawed sea bass (Dicentrarchus labrax) fillets using the two-dimensional polyacrylamide gel electrophoresis (2-DE) technique. Distinct gel patterns, due to proteins with low molecular weight and low isoelectric points, distinguished fresh fillets from frozen–thawed ones. Frozen–thawed fillets showed two specific protein spots as early as the first day of the study. However, these spots were not observed in fresh fillets until at least 13days of storage between 0 and 4°C, fillets were judged, beyond this period, fish were unfit for human consumption as revealed by complementary studies on fish spoilage indicators namely total volatile basic nitrogen and biogenic amines. Mass spectrometry identified the specific proteins as parvalbumin isoforms. Parvalbumins may thus be useful markers of differentiation between fresh and frozen–thawed sea bass fillets.

Interleukin-18 alters protein expressions of neurodegenerative diseases-linked proteins in human SH-SY5Y neuron-like cells.

Chronic inflammation and oxidative stress (OS) are present in Alzheimer's disease (AD) brains in addition to neuronal loss, Amyloid-β (Aβ) plaques and hyperphosphorylated tau-protein neurofibrillary tangles (NFTs). Previously we showed that levels of the pro-inflammatory cytokine, interleukin-18 (IL-18), are elevated in post-mortem AD brains. IL-18 can modulate the tau kinases, Cdk5 and GSK3β, as well as Aβ-production. IL-18 levels are also increased in AD risk diseases, including type-2 diabetes and obesity. Here, we explored other IL-18 regulated proteins in neuron-like SH-SY5Y cells. Differentiated SH-SY5Y cells, incubated with IL-18 for 24, 48, or 72 h, were analyzed by two-dimensional gel electrophoresis (2D-DIGE). Specific altered protein spots were chosen and identified with mass spectrometry (MS) and verified by western immunoblotting (WIB). IL-18 had time-dependent effects on the SH-SY5Y proteome, modulating numerous protein levels/modifications. We concentrated on those related to OS (DDAH2, peroxiredoxins 2, 3, and 6, DJ-1, BLVRA), Aβ-degradation (MMP14, TIMP2), Aβ-aggregation (Septin-2), and modifications of axon growth and guidance associated, collapsin response mediator protein 2 (CRMP2). IL-18 significantly increased antioxidative enzymes, indicative of OS, and altered levels of glycolytic α- and γ-enolase and multifunctional 14-3-3γ and -ε, commonly affected in neurodegenerative diseases. MMP14, TIMP2, α-enolase and 14-3-3ε, indirectly involved in Aβ metabolism, as well as Septin-2 showed changes that increase Aβ levels. Increased 14-3-3γ may contribute to GSK3β driven tau hyperphosphorylation and CRMP2 Thr514 and Ser522 phosphorylation with the Thr555-site, a target for Rho kinase, showing time-dependent changes. IL-18 also increased caspase-1 levels and vacuolization of the cells. Although our SH-SY5Y cells were not aged, as neurons in AD, our work suggests that heightened or prolonged IL-18 levels can drive protein changes of known relevance to AD pathogenesis.

Serum differential protein identification of Xinjiang Kazakh esophageal cancer patients based on the two-dimensional liquid-phase chromatography and LTQ MS

The aim of this study was to investigate the impact of chemo-radiotherapy on serum protein expression of the esophageal cancer patients and discover potential biomarkers by detecting serum proteins mass spectrometry of the healthy Kazakh people in Xinjiang as well as the patients before and after their chemo-radiotherapy. In order to separate and compare the three serum samples (the healthy group's, the patients' before and after chemo-radiotherapy) with two-dimensional protein liquid chromatography system (Proteome LabTM PF-2D), then detect the differential protein spots with linear trap quadruple mass spectrometer (LTQ MS/MS). (1) The Kazakh esophageal cancer patients got 21 expressed protein spots peaks with significant difference after chemo-radiotherapy compared with before; before the treatment there were 10 different expressed protein spots compared with the healthy group, and after it there were four peaks in the expression of protein spots compared with the healthy group. (2) After LTQ mass spectrometric detection, 22 proteins were up-regulated in serum samples of the healthy group, 22 were up-regulated of the patients before medical treatment and 5 were up-regulated after chemo-radiotherapy. (3) 8 proteins including APOA1 can be served as serum markers in Kazakh esophageal cancer diagnosis, and proteins like CLU can be served as serum markers in judging the resistance and sensitivity towards chemo-radiotherapy. (4) The abnormal expressions of APOC2, APOC3, Antithrombin-III in esophageal cancer were discovered for the first time. Specific protein spots related to Xinjiang Kazakh esophageal cancer diagnosis and chemo-radiotherapy can be identified in the serum, which will probably become a maker in Kazakh esophageal cancer diagnosis and therapeutic evaluation.

Proteomic Analysis of Testicular Ischemia-Reperfusion Injury in Rats

ABSTRACTTesticular torsion is a urological emergency that leads to serious testicular damage and male infertility. We performed this study to identify specific proteins that are differentially expressed in response to testicular torsion and detorsion-induced ischemia-reperfusion (I-R) injury. Adult male rats were divided into two groups: a sham-operated group and a testicular I-R group. Testicular torsion was induced by rotating the left testis 720° in a clockwise direction for 1 hr, and then, detorsion was performed for 24 hr. After this testicular tissues were collected, protein analysis was performed using two-dimensional gel electrophoresis and Western blot analyses. Testicular I-R injury resulted in serious histopathologic damage to the germinal cells in the seminiferous tubules and increased the number of TUNEL-positive cells in testicular tissue. Specific protein spots with a greater than 2.5-fold change in intensity between the sham-operated and testicular I-R groups were identified by mass spectrometry. Among these proteins, levels of peroxiredoxin 6, thioredoxin, heterogeneous nuclear ribonucleoproteins, ubiquitin carboxyl terminal hydrolase isozyme L5 and zinc finger AN1-type domain 3 were decreased in the testicular I-R group compared to the sham-operated group. Moreover, Western blot analysis clearly showed the decrease of these proteins in the testicular I-R group. These proteins have spermatogenesis and anti-oxidative functions. These findings suggest that testicular I-R results in cell death due to altered expression of several proteins with spermatogenesis and anti-oxidation functions.

Differential Proteomic Analysis of Carbon Ion Radiation in Sheep Sperm

This study is first to investigate proteomic changes in sheep sperm induced by carbon ion radiation using two-dimensional electrophoresis (2-DE) analysis in the project of breeding a new variety of sheep. Differential expression proteins were detected using the PDQuest 8.0 software after staining with Coomassie blue. Valid spots were then analyzed through liquid chromatography tandem mass spectrometry (LC-MS/MS). Among the 480 total protein spots displayed in 2-D gels, 6 specific protein spots were observed in sperm gels. A search against protein sequences in the National Center for Biotechnology Information databases (NCBI) indicated that differentially expressed proteins correspond to two proteins, identified to be enolase and transcription factor AP-2-alpha (TFAP-2α). The two proteins were up-regulated in the irradiated sperm. To the best of our knowledge, this study is the first to identify proteomic changes induced by carbon ion radiation in sheep sperm. The analysis of differential expression protein may be useful in identifying new breeding markers in sheep reproduction and in clarifying the mechanisms involved in irradiation or space breeding.

Changes in inflorescence protein during advanced stages of floret development in Buchloe dactyloides (Poaceae)

Buffalograss, Buchloe dactyloides, is a dioecious species native to the Great Plains of North America. The florets at the early stages of development possess both gynoecium and androecium organ primordia but later become unisexual. Very little is known about the proteomic changes that occur when the florets change from hermaphroditism to unisexuality. We compared the protein composition of florets at the hermaphroditic stage with that at the unisexual stage. The development stage of the floret was determined by stereomicroscopic observation. Two-dimensional gel electrophoresis was used to separate the proteins extracted from female and male inflorescences. Stage- specific protein maps, with an average of about 400 spots per map, were analyzed with the protein analysis software. Eighteen spots were found to be differentially expressed between the hermaphrodite and unisexual stages. Of these, 12 were present at both stages but with a different expression value. Four specific spots appeared at the hermaphrodite stage and disappeared at the unisexual stage. Two specific protein spots were associated with female and male floret differentiation. One appears to be associated with contabescence in the female floret and the final protein appears to lead to the abortion of gynoecium in the male floret. The MALDI TOF/TOF technique was used for peptide mass fingerprinting of the differentially expressed proteins and the MASCOT software was used to search the protein database. However, only two protein spots were identified from the database. These were aldolase1 and Os05g0574400 (similar to malate dehydrogenase). This type of proteomic study can help to identify novel protein products and determine the mechanisms involved in the floral sex differentiation process in buffalo grass.

스트레스 음파 처리에 따른 아메리카잎굴파리(Liriomyza trifolii)의 생리 변화와 프로테오믹 분석

본 연구는 음파 처리에 따른 아메리카잎굴파리(Liriomyza trifolii)의 생리 변화를 발육 시기별로 분석하였다. 유충의 섭식량은 섭식을 통해 형성된 잎 내부 갱도의 길이로 분석했다. 이때 5,000 Hz (95 dB)의 음파 처리는 섭식 행동을 크게 둔화시켰다. 이러한 스트레스 음파는 또한 용발육에 영향을 주어 315 - 5,000 Hz 음파 범위에서 발육 지연 효과를 나타냈고, 1,000 - 5,000 Hz 음파 범위에서 우화를 억제하였다. 스트레스 음파는 암컷의 산란 행동을 억제시켰으나 성충의 주광성 행동에는 영향을 주지 않았다. 스트레스 음파(5,000 Hz, 95 dB)를 받은 용은 이차원 전기영동 분석 결과 단백질 발현 양상에서 무처리구와 뚜렷한 차이를 보였다. 특정 단백질에 대한 MALDI-TOF 분석 결과는 trafficking protein particle complex I, triosephosphate isomerase, hypothetical protein TcasGA2_TC013388, polycystin-2, paraneoplastic neuronal antigen MA1 및 tropomyosin I (isoform M)의 단백질들이 무처리구에서 발견되고, 음파 처리구에서는 소멸되었다. 반면에 DOCK9, cytoskeletal keratin II 및 <TEX>$F_0F_1$</TEX>-ATP synthase beta subunit은 스트레스 음파 처리에 따라 새롭게 나타나는 단백질로 판명되었다. 이러한 스트레스 음파는 아메리카잎굴파리의 살충제에 대한 감수성을 크게 증가시켰다. 이상의 결과는 아메리카잎굴파리의 생리 과정을 교란하는 스트레스 음파가 작용한다는 것을 제시하고 있다. 또한 본 연구는 이러한 스트레스 음파를 이용하여 아메리카잎굴파리에 대한 새로운 물리적 해충 방제 기술 개발 가능성을 제시하고 있다. This study investigated the adverse effects of sound treatment on physiological processes of the American leafminer, Liriomyza trifolii, during several developmental stages. Larval feeding activity was analyzed by measuring feeding tunnel length. It was significantly suppressed by sound treatment (5,000 Hz, 95 dB). Sound treatment delayed the pupal period at 315 - 5,000 Hz and prevented adult emergence at 1,000 - 5,000 Hz. Female oviposition was also inhibited by the stress sound treatments. However, phototactic adult movement was not affected by sound treatment. Pupae treated with 5,000 Hz showed marked changes in protein patterns analyzed by two dimensional electrophoresis. MALDI-TOF analysis of specific protein spots indicated that trafficking protein particle complex I, triosephosphate isomerase, hypothetical protein TcasGA2_TC013388, polycystin-2, paraneoplastic neuronal antigen MA1, and tropomyosin I (isoform M) were predicted in the control insects and disappeared in the insects treated with sound. By contrast, DOCK9, cytoskeletal keratin II, and F0F1-ATP synthase beta subunit were predicted only in the sound-treated insects. Furthermore, stress sound significantly increased the susceptibility of L. trifolii to insecticides. These results suggest that physiological processes of L. trifolii are altered by sound stress, which may be exploited to develop a novel physical control tactic against L. trifolii.