• Title/Summary/Keyword: stress gene

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Transgenic Tobacco Plants Expressing the Bacterial Levansucrase Gene Show Enhanced Tolerance to Osmotic Stress

  • Park, Jeong-Mee;Kwon, Suk-Yoon;Song, Ki-Bang;Kwak, Ju-Won;Lee, Suk-Bae;Nam, Young-Woo;Shin, Jeong-Sheop;Park, Young-In;Rhee, Sang-Ki;Paek, Kyung-Hee
    • Journal of Microbiology and Biotechnology
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    • v.9 no.2
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    • pp.213-218
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    • 1999
  • Fructans are polyfructose molecules that function as nonstructural storage carbohydrates in several plants. In addition, it has been suggested that, due to their solubility, they can play an important role in helping plants survive periods of osmotic stress. In order to study the effect of levan synthesis on plant growth, the coding region of the levansucrase gene, which was isolated from Zymomonas mobilis, was introduced into tobacco plants using Agrobacterium tumefaciens-mediated transformation. The presence of the levansucrase gene in transgenic plants was verified by genomic DNA gel blot analysis. RNA gel blot and immunoblot analyses showed an accumulation of the corresponding transcript and protein product of the bacterial levansucrase gene in transgenic plants. Furthermore, a thin layer chromatography analysis revealed that fructans were synthesized and deposited in transgenic tobacco plants. When $T_1$ seeds were germinated and grown under polyethylene glycol-mediated drought stress or cold stress, the transgenic seedlings displayed a substantially higher level of growth than that of untransformed plants. These results suggest that fructans may playa significant role in the tolerance of plants under osmotic stress.

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Cloning and Characterization of the HSP70 Gene, and Its Expression in Response to Diapauses and Thermal Stress in the Onion Maggot, Delia antiqua

  • Chen, Bin;Kayukawa, Takumi;Monteiro, Antonia;Ishikawa, Yukio
    • BMB Reports
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    • v.39 no.6
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    • pp.749-758
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    • 2006
  • The cytosolic members of the HSP70 family of proteins play key roles in the molecular chaperone machinery of the cell. In the study we cloned and sequenced the full-length cDNA of Delia antiqua HSP70 gene, which is 2461 bp long and encodes 643 a.a. with a calculated molecular mass of 70,787 Da. We investigated gene copies of cytosolic HSP70 members of 4 insect species with complete genome available, and found that they are quite variable with species. In order to characterize this protein we carried out an alignment and a phylogenetic analysis with 41 complete protein sequences from insects. The analysis divided the cytosolic members of the family into two classes, HSP70 and HSC70, distinguishable on the basis of 15 residues. HSP70 class members were slightly shorter in length and smaller in molecular mass relative to the HSC70 class members, and the conservative and functional regions in these sequences were documented. Mainly, we investigated the expression of Delia antiqua HSP70 gene, in response to diapauses and thermal stresses. Both summer and winter diapauses elevated HSP70 transcript levels. Cold-stress led to increased HSP70 expression levels in summer- and winter-diapausing pupae, but heat-stress elevated the levels only in the winter-diapausing pupae. In all cases, the expression levels, after being elevated, gradually decreased with time. HSP70 expression was low in non-diapausing pupae but was up-regulated following cold- and heat-stresses. Heat-stress gradually increased the mRNA level with time whereas cold-stress gradually decreased levels after an initial increase.

Overexpression of Heat Shock Factor Gene HsfA3 Increases Galactinol Levels and Oxidative Stress Tolerance in Arabidopsis

  • Song, Chieun;Chung, Woo Sik;Lim, Chae Oh
    • Molecules and Cells
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    • v.39 no.6
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    • pp.477-483
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    • 2016
  • Heat shock factors (Hsfs) are central regulators of abiotic stress responses, especially heat stress responses, in plants. In the current study, we characterized the activity of the Hsf gene HsfA3 in Arabidopsis under oxidative stress conditions. HsfA3 transcription in seedlings was induced by reactive oxygen species (ROS), exogenous hydrogen peroxide ($H_2O_2$), and an endogenous $H_2O_2$ propagator, 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone (DBMIB). HsfA3-overexpressing transgenic plants exhibited increased oxidative stress tolerance compared to untransformed wild-type plants (WT), as revealed by changes in fresh weight, chlorophyll fluorescence, and ion leakage under light conditions. The expression of several genes encoding galactinol synthase (GolS), a key enzyme in the biosynthesis of raffinose family oligosaccharides (RFOs), which function as antioxidants in plant cells, was induced in HsfA3 overexpressors. In addition, galactinol levels were higher in HsfA3 overexpressors than in WT under unstressed conditions. In transient transactivation assays using Arabidopsis leaf protoplasts, HsfA3 activated the transcription of a reporter gene driven by the GolS1 or GolS2 promoter. Electrophoretic mobility shift assays showed that GolS1 and GolS2 are directly regulated by HsfA3. Taken together, these findings provide evidence that GolS1 and GolS2 are directly regulated by HsfA3 and that GolS enzymes play an important role in improving oxidative stress tolerance by increasing galactinol biosynthesis in Arabidopsis.

Salt-induced Differential Gene Expression in Italian Ryegrass (Lolium multiflorum Lam.) Revealed by Annealing Control Primer Based GeneFishing approach

  • Lee, Ki-Won;Lee, Sang-Hoon;Choi, Gi Jun;Ji, Hee Jung;Hwang, Tae Young;Kim, Won Ho;Rahman, Md. Atikur
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.37 no.3
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    • pp.231-236
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    • 2017
  • Salt stress is one of the most limiting factors that reduce plant growth, development and yield. However, identification of salt-inducible genes is an initial step for understanding the adaptive response of plants to salt stress. In this study, we used an annealing control primer (ACP) based GeneFishing technique to identify differentially expressed genes (DEGs) in Italian ryegrass seedlings under salt stress. Ten-day-old seedlings were exposed to 100 mM NaCl for 6 h. Using 60 ACPs, a total 8 up-regulated genes were identified and sequenced. We identified several promising genes encoding alpha-glactosidase b, light harvesting chlorophyll a/b binding protein, metallothionein-like protein 3B-like, translation factor SUI, translation initiation factor eIF1, glyceraldehyde-3-phosphate dehydrogenase 2 and elongation factor 1-alpha. These genes were mostly involved in plant development, signaling, ROS detoxification and salt acclimation. However, this study provides new molecular information of several genes to understand the salt stress response. These genes would be useful for the enhancement of salt stress tolerance in plants.

Validation of Reference Genes for Quantifying Changes in Physiological Gene Expression in Apple Tree under Cold Stress and Virus Infection (저온과 바이러스 감염에 노출된 사과나무의 생리적 유전자 정량 측정용 유전자들의 발현 분석 및 검증)

  • Yoon, Ju-Yeon;Jeong, Jae-Hoon;Choi, Seung-Kook
    • Research in Plant Disease
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    • v.26 no.3
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    • pp.144-158
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    • 2020
  • Quantitative reverse transcription PCR is used for gene expression analysis as the accurate and sensitive method. To analyze quantification of gene expression changes in apple plants, 10 housekeeping genes (ACT, CKL, EF-1α, GAPDH, MDH, PDI, THFs, UBC, UBC10, and WD40) were evaluated for their stability of expression during infection by Apple stem grooving virus (ASGV) or in cold-stress apple plant buds. Five reference-gene validation programs were used to establish the order of the most stable genes for ASGV as CKL>THFs>GAPDH>ACT, and the least stable genes WD40CKL>UBC10, and the least stable genes were ACT

Induction of a Sweetpotato Anion Peroxidase swpa2 Gene Expression by Stress-related Chemicals and Pectobacterium chrysanthemi (스트레스 관련 화합물 처리 및 병원균 감염에 의한 고구마 산성 퍼옥시다제 swpa2 발현 유도)

  • Kim, Yun-Hee;Ryu, Sun-Hwa;Kim, Kee-Yeun;Kwon, Suk-Yoon;Bang, Jae-Wook;Kwak, Sang-Soo
    • Journal of Plant Biotechnology
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    • v.31 no.1
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    • pp.83-88
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    • 2004
  • Expression of an anionic peroxidase swpa2 gene isolated from cultured cells of sweetpotato (Ipomoea batatas) was investigated under various stress conditions by RT-PCR. The swpa2 gene was not expressed in any tissues of intact sweetpotato plant grown at the normal condition. The expression of this gene was strongly induced in leaf tissue by treatment of $H_2O$$_2$ (440mM). Treatment of NaCl (100mM), ABA (0.1mM) and methyl jasmonate(MeJA, 0.1mM) also induced the expression of swpa2 gene. Interestingly, salicylic acid (SA, 0.1 mM) did not induce the expression of swpa2 gene, indicating that anionic swpa2 POD is differently involved in SA and MeJA signaling pathways. In addition, swpa2 gene was strongly induced in sweetpoato leaf tissues infected with Pectobacterium chrysanthemi, indicating that swpa2 is involved in defense related to the pathogenesis of P. chrysanthemi in sweetpotato plants. These results strongly suggest that swpa2 gene is involved in overcoming oxidative stresses caused by both abiotic and biotic stress.

Comparative transcriptome analysis of heat stress responsiveness between two contrasting ginseng cultivars

  • Jayakodi, Murukarthick;Lee, Sang-Choon;Yang, Tae-Jin
    • Journal of Ginseng Research
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    • v.43 no.4
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    • pp.572-579
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    • 2019
  • Background: Panax ginseng has been used in traditional medicine to strengthen the body and mental well-being of humans for thousands of years. Many elite ginseng cultivars have been developed, and ginseng cultivation has become well established during the last century. However, heat stress poses an important threat to the growth and sustainable production of ginseng. Efforts have been made to study the effects of high temperature on ginseng physiology, but knowledge of the molecular responses to heat stress is still limited. Methods: We sequenced the transcriptomes (RNA-Seq) of two ginseng cultivars, Chunpoong (CP) and Yunpoong (YP), which are sensitive and resistant to heat stress, respectively, after 1- and 3-week heat treatments. Differential gene expression and gene ontology enrichment along with profiled chlorophyll contents were performed. Results: CP is more sensitive to heat stress than YP and exhibited a lower chlorophyll content than YP. Moreover, heat stress reduced the chlorophyll content more rapidly in CP than in YP. A total of 329 heat-responsive genes were identified. Intriguingly, genes encoding chlorophyll a/b-binding proteins, WRKY transcription factors, and fatty acid desaturase were predominantly responsive during heat stress and appeared to regulate photosynthesis. In addition, a genome-wide scan of photosynthetic and sugar metabolic genes revealed reduced transcription levels for ribulose 1,5-bisphosphate carboxylase/oxygenase under heat stress, especially in CP, possibly attributable to elevated levels of soluble sugars. Conclusion: Our comprehensive genomic analysis reveals candidate loci/gene targets for breeding and functional studies related to developing high temperature-tolerant ginseng varieties.

Cloning, Characterization, and Functional Analysis of Maize DEHYDRIN2

  • Paek, Nam-Chon;Jung, Hun-Ki
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.47 no.2
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    • pp.116-122
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    • 2002
  • Dehydrins (LEA Dll proteins) are one of the typical families of plant proteins that accumulate in response to dehydration, cold stress, abscisic acid, or during seed maturation. A 1.3-kb cDNA was cloned from a cDNA expression library of 5-day-old germinating maize scutellums under drought stress. The deduced protein sequence indicated a dehydrin gene encoding SK$_3$ LEA protein typically expressed during cold acclimation, but not by drought stress in barley and wheat. Thus, it was named maize DEHYDRIN2 (ZmDhn2). It accumulates rapidly and highly in drought-stressed scutellum and leaf tissues at any stage, but not under cold stress. ZmDhn2 gene was transformed into Arabidopsis thaliana for functional analysis under drought condition. From electrolyte leakage test, no significant difference showed between wild type and transformants under normal growth condition, but the leakage level of electrolyte in wild type plants was about 3 times as high as that in the transformed plants under drought stress. It suggests that ZmDHN2 playa role in increasing drought tolerance.

Transcriptome analysis of internal and external stress mechanisms in Aster spathulifolius Maxim.

  • Sivagami, Jean Claude;Park, SeonJoo
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.04a
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    • pp.35-35
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    • 2019
  • Aster spathulifolius Maxim. is belongs to the Asteraceae family which is distributed only in Korea and Japan. It is recognize as a traditionally medicinal plants and economically valuable in ornamental field. However, among the Asteraceae family, the Aster genus, which is lacks in genomic resources and information of molecular function. Therefore, we used high throughput RNA-sequencing transcriptome data of the A. spathulifolius to know molecular level function. DeNovo assembly produced 98,660 unigene with N50 value 1126 bp. Unigenes was performed to analyses the functional annotation against NCBI database like plant database of nucleotide (Nt) and non-redundant protein (Nr), Pfam, Uniprot, KEGG and Transcriptional factor (TF). In addition, Distribution of SSR markers also analyzed for future perfectives. Further, Comparing with other two Asteraceae family species like, Karelinia caspica and Chrysanthemum morifolium to the A. spathulifolius shows the number of gene that regulated in internal and external stress respectively salt-tolerant and heat and drought stress to understand the molecular basis related to the different environments stress.

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Lycopersicon Eculentum C2H2-type Zinc Finger Protein Induced by Oxidative Stress Especially

  • Seong, Eun-Soo;Lee, Ji-Yeon;Yu, Chang-Yeon;Yang, Deok-Chun;Eom, Seok-Hyun;Cho, Dong-Ha
    • Journal of Plant Biotechnology
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    • v.34 no.3
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    • pp.167-172
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    • 2007
  • A tomato zinc-finger protein gene, LeZFP1, encoding the Cys2/His2-type zinc-finger transcription factor was searched from cDNA microarray analysis of gene expression following induction of the overexpressed tomato transgenic plants showing resistance for pathogen and abiotic stresses. The full-length cDNA of LeZFP1 encoded a protein of 261 amino acid residues. Analysis of the deduced amino acid sequence of LeZFP1 revealed that it shares high sequence identity with pepper CAZFP1 (81% identity). We found that single copy of LeZFP1 gene is present in the tomato genome through southern blot analysis. The LeZFP1 transcripts were constitutively expressed in the tomato mature and young leaves, but were detectable weakly in the flower, stem and root. The LeZFP1 transcripts were significantly reduced in treated leaf tissues with NaCl and mannitol. The LeZFP1 gene was induced by oxidative stress especially. Our results indicated that LeZFP1 may play a role function involved in oxidative stress signaling pathways.