Morphologic and anatomic response, catalase gene expression in drought varieties of tomato and bioinformatics analyses of microarray studies of the catalase gene

2021 ◽  
Author(s):  
Hanieh Mohajjel Shoja ◽  
Taha Khezriani ◽  
Maryam Kolahi ◽  
Elham Elham Mohajel Kazemi ◽  
Milad Yazdi
Keyword(s):  
Gene Expression ◽  
Drought Stress ◽  
Real Time ◽  
Morphological Changes ◽  
Field Capacity ◽  
Bioinformatics Analyses ◽  
Catalase Gene ◽  
Significant Difference ◽  
Drought Conditions ◽  
Microarray Studies

Abstract Crops in arid and semi-arid regions are exposed to adverse environmental factors such as drought. Experiments were conducted to determine the morphologic and anatomic response of drought-susceptible and tolerant varieties of tomato (Solanum lycopersicum L.) under drought conditions (100%, 75%, 50%, 25% of field capacity). To investigate the role of antioxidant enzyme, catalase gene expression was examined by real-time RT-qPCR and microarray studies of the catalase gene in tomatoes under stress examined utilizing bioinformatics. The results showed significant morphological changes under drought conditions. Anatomical studies revealed that CaljN3 is more resistant than SuperstrainB varieties under drought stress. Relative expression of the CAT1 gene did not show any significant difference in both Caljn3 and SuperstrainB varieties based on quantitative Real-Time PCR, under drought stress. The bioinformatics results from microarray analysis revealed that this gene did not show a significant difference in expression in any of the cultivars and under any of the stresses. This gene is in the conserve cluster, a cluster with 118 members and a z score of 14.26148. This showed that this cluster is fully protected between two susceptible and tolerant varieties. The enrichment gene of this cluster did not show any significant intracellular pathways. It appears that in response to stress, an activating mechanism other than catalase is necessary. The fight against oxidative stress may begin one step before that of the enzymes and seeks to combat the stressor by activating proteins, especially channels, pumps and some cellular messengers.

Respon Morfologis dan Ekspresi Gen Aquaporin pada Padi IR 64 yang Mengalami Cekaman Kekeringan pada Fase Reproduktif (Morphological Responses and Aquaporin Gene Expression in Rice IR64 under Drought Stress at the Reproductive Stage)

2018 ◽  
Vol 7 (2) ◽  
Author(s):  
Made Pharmawati ◽  
Ni Nyoman Wirasiti ◽  
Luh Putu Wrasiati
Keyword(s):  
Gene Expression ◽  
Drought Stress ◽  
Real Time ◽  
Real Time Pcr ◽  
Control Plant ◽  
Reproductive Stage ◽  
Limiting Factors ◽  
Rice Grain ◽  
Aquaporin Gene ◽  
Peg Treatment

Abstrak Cekaman kekeringan merupakan faktor pembatas penting bagi pertumbuhan dan produktivitas tanaman termasuk padi.      Penelitian ini bertujuan menganalisis respon padi IR64 terhadap cekaman kekeringan dengan pemberian polietilen glikol (PEG) pada fase reproduktif.  Penelitian juga bertujuan menganalisis ekspresi gen aquaporin akibat cekaman kekeringan.  Bibit padi ditanam dalam pot dan perlakuan PEG dengan konsentrasi 108g/L (-0.25MPa) dan 178g/L (-0.52 MPa) diberikan saat munculnya panikula. Perlakuan diberikan selama 2 minggu, kemudian tanaman disiram kembali.  Ekspresi gen diamati pada akhir perlakuan dengan semi kuantitatif real time PCR.  Ekstraksi RNA menggunakan RNeasy plant mini kit, sedangkan sintesis cDNA menggunakan Transcriptor First Strand cDNA Kit.  Hasil penelitian menunjukkan bahwa jumlah malai dan berat total malai berkurang akibat cekaman kekeringan.  Persentase gabah kosong mencapai 84,6% pada perlakuan PEG-0,52 MPa, sedangkan pada perlakuan PEG -0,25 MPa persentase gabah kosong sebesar 67,8%.  Pada kontrol persentase gabah kosong adalah 10,3%.  Ekspresi gen OsPIP2;7 sedikit menurun pada perlakuan PEG -0,52 MPa.Kata kunci: ekspresi gen, IR64, kekeringan, padi, PEG  Abstract Drought stress is one of the limiting factors of plant growth and productivity including rice.  The aim of this study was to analyze responses of IR64 rice to polyethylene glycol (PEG)-induced-drought stress at the reproductive stage.  This study also aimed to analyze the expression of aquaporin under drought stress.  Rice seedlings were grown in pot system and PEG treatment at concentration of -0.25MPa (108g/L) and -0.52 MPa (178g/L) were given when the panicles arose.  Treatments were conducted for 2 weeks, after that the plants were rewatered.  Gene expression was evaluated at the end of PEG treatment using semi quantitative real time PCR. RNA was extracted using RNeasy plant mini kit, while cDNA synthesis was done using Transcriptor First Strand cDNA Kit.  The results showed that the number and weight of rice ear were less in plant treated with PEG than in control.  The percentage of empty rice grain reached 84.6% at PEG -0.52 MPa, while at PEG -0.25 MPa the percentage of empty grain was 67.8%.  In control plant, the percentage of empty grain was 10.3%.  Drought stress did not alter the expression of OsPIP2;7.  Keywords: drought, gene expression, IR64, PEG, rice

scholarly journals Differential Activity of Antioxidant Enzymes and Physiological Changes in Wheat (Triticum aestivum L.) Under Drought Stress

2019 ◽  
Vol 11 (2) ◽  
pp. 266-276
Author(s):  
Kamal MIRI-HESAR ◽  
Ali DADKHODAIE ◽  
Saideh DOROSTKAR ◽  
Bahram HEIDARI
Keyword(s):  
Water Stress ◽  
Drought Stress ◽  
Field Capacity ◽  
Triticum Aestivum L ◽  
Plant Production ◽  
Breeding Programs ◽  
Drought Tolerant ◽  
Severe Water Stress ◽  
Significant Difference ◽  
Relative Water

Drought stress is one of the most significant environmental factors restricting plant production all over the world. In arid and semi-arid regions where drought often causes serious problems, wheat is usually grown as a major crop and faces water stress. In order to study drought tolerance of wheat, an experiment with 34 genotypes including 11 local and commercial cultivars, 17 landraces, and six genotypes from International Maize and Wheat Improvement Center (CIMMYT) was conducted at the experimental station, School of Agriculture, Shiraz University, Iran in 2010-2011 growing season. Three different irrigation regimes (100%, 75% and 50% Field Capacity) were applied and physiological and biochemical traits were measured for which a significant difference was observed in genotypes. Under severe water stress, proline content and enzymes’ activities increased while the relative water content (RWC) and chlorophyll index decreased significantly in all genotypes. Of these indices, superoxide dismutase (SOD) and RWC were able to distinguish tolerant genotypes from sensitives. Moreover, yield index (YI) was useful in detecting tolerant genotypes. The drought susceptibility index (DSI) varied from 0.40 to 1.71 in genotypes. These results indicated that drought-tolerant genotypes could be selected based on high YI, RWC and SOD and low DSI. On the whole, the genotypes 31 (30ESWYT200), 29 (30ESWYT173) and 25 (Akbari) were identified to be tolerant and could be further used in downstream breeding programs for the improvement of wheat tolerance under water limited conditions.

Real-Time PCR Gene Expression Profiling of Microarray Gene Signatures in Acute Leukemia.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4401-4401
Author(s):  
Ebrahim Sakhinia ◽  
Mahboubeh Farahangpour ◽  
John A. Liu Yin ◽  
Gerard Brady ◽  
Judith A. Hoyland ◽  
...  
Keyword(s):  
Gene Expression ◽  
Real Time ◽  
Real Time Pcr ◽  
Cyclin D3 ◽  
Expression Level ◽  
Tumor Type ◽  
Gene Signatures ◽  
Significant Difference ◽  
The Mean ◽  
Microarray Gene

Abstract Cancer subtype discovery and classification using microarray gene signatures has the potential to transform pathological diagnosis but measurement of indicator genes in routine practice remains difficult. We tested use of real-time PCR measurement of indicator genes for AML and ALL (Golub et al, Science, 1999) as a method for validation and application of microarray gene signatures. Mononuclear cells (MC) were isolated from whole bone marrow (BM) aspirates by density gradient centrifugation and sorted into unselected (total), CD34+ve and CD34-ve fractions. The mRNA in each fraction was globally amplified using a PolyA PCR method. We measured the expression profile of the 17 top ranked genes (cystatin C, leptin receptor, fumarylacetoacetate, CD33, HoxA9, adipsin, proteoglycan 1, LTC4 synthase, LYN, C-myb, MB-1, cyclin D3, SNF2, RbAp48, proteasome iota, HkrT-1 and E2A) from Golub et al (1999) by real-time PCR. All values were calibrated against control standards and normalized to the mean of three housekeeping genes (IF2-beta, GAPDH and human ribosomal protein S9). Data for all 17 genes were obtained for 4 (ALL), 26 (AML), 12 (AML remission) and 9 (morphologically normal) BM samples, each fractionated into three fractions (total MC, CD34+ve MC & CD34−ve MC). There was no significant difference in the mean of three housekeeping gene expression levels between the diagnostic groups. Comparison of the expression level of the other genes confirmed ability to separate AML and ALL, whilst the direction of expression change (increased or decreased) for each gene between AML and ALL was the same as found by Golub et al. In particular, c-myb showed largest significant increase in ALL vs AML in the total BM fraction, whilst cystain c was increased in AML in the CD34−ve fraction. hSNF2b was significantly increased in the ALL total B.M fraction and Hox-A9 was significantly increased in the AML CD34+ve B.M fraction. Furthermore expression level of LYN and CD33 was significantly increased in AML compared to remission AML, indicating ability of the method to determine activity status of disease. In addition, several of the genes provided better separation between AML and ALL when measured in the CD34+ve and −ve fractions indicating more prominent expression in cells of different maturity and that prior fractionation is diagnostically more informative. The results demonstrate ability of the method to validate gene expression signatures by an independent method, which is simple, sensitive and robust, allowing translation to routine clinical use. Whilst the present study used AML and ALL, in principle the method could be extended to any other tumor type for which gene signatures exist.

scholarly journals Comparison of Glutathione Reductase Gene Expression in Drought-sensitive and Tolerant Soybean Lines using Real-time PCR

2021 ◽  
Author(s):  
Mohammad Hassanvand ◽  
Shahab Khaghani ◽  
Mahdi Changizi ◽  
Masoud Gomarian ◽  
Ezatollah Sedaghatfar
Keyword(s):  
Gene Expression ◽  
Abiotic Stress ◽  
Drought Stress ◽  
Plant Growth ◽  
Glutathione Reductase ◽  
Real Time ◽  
Real Time Pcr ◽  
High Rate ◽  
Drought Tolerant ◽  
Reductase Gene

Abstract Background: Global climate change and associated adverse abiotic stress conditions, such as drought, salinity, heavy metals, waterlogging, extreme temperatures, oxygen deprivation, etc., greatly influence plant growth and development, ultimately affecting crop yield and quality, as well as agricultural sustainability in general. This study provides new insights into the analysis of the function of soybean genes in abiotic stress. Drought is one of the significant constraints that limit agricultural productivity. Some factors, including climate changes and acreage expansion, indicate the need for developing drought-tolerant Genotypes.Materials and methods: The study of the expression Glutathione Reductase (GR) gene in soybean drought-tolerant and sensitive cultivars using real-time PCR. Seeds from (drought-sensitive) and (drought-tolerant) lines were planted under specific temperature conditions drought stress treatment, in the research greenhouse of Islamic Azad University of Arak, Iran. Changes in gene expression compared to reference genes were recorded using the formula 2-ΔΔCT. Three technical replications were given for each cDNA sample related to each sampling and used to analyze test data from MINITAB16 software.Results: The results showed that the threshold expression of gene expression (Glutathione) in the Pyramid line had the highest expression of drought resistance and the lowest expression of the Glutathione Reductase gene belonging to the Will line. The theory is also true that chaperone proteins produced during the plant growth cycle are not destroyed to express the Glutathione Reductase gene. The expression cycle of the Glutathione Reductase gene shows that the proteins produced by this gene have a high rate of expression and increase in cell drought stress. This gene expression continues until the pressure ends. The results showed that lines and cultivars with a weak expression against drought stress could have a high expression at the beginning of drought stress but a decrease in gene expression rate during stress. Drought stress-sensitive lines have a decreasing expression in the middle and end of stress during the stress period.

scholarly journals THE EFFECTS OF POTASSIUM APPLICATIONS ON DROUGHT STRESS IN SUGAR BEET: PART II. PLANT NUTRITION CONTENT

2020 ◽  
Vol 4 (3) ◽  
pp. 203-216
Author(s):  
Gizem AKSU ◽  
Hamit ALTAY
Keyword(s):  
Drought Stress ◽  
Sugar Beet ◽  
Plant Nutrition ◽  
Critical Role ◽  
Calcium Content ◽  
Field Capacity ◽  
Natural Phenomenon ◽  
Water Conditions ◽  
Drought Conditions ◽  
Potassium Application

This is the second in a series of papers describing the effects of potassium applications on drought stress in sugar beet. Drought is a natural phenomenon that can affect water resources and agriculture. In this research, the effect of potassium applications under drought stress on some plant nutrition of sugar beet, which is a strategic plant, was investigated. In the experiment, irrigation levels were kept at 33%, 66% and 100% of field capacity. Different doses (10-20-40-80 mg kg-1) of potassium were applied to the plants. The plants were grown in the growth chamber under controlled conditions (day/night 16/8 hours, 25/15 0C, 60-70% humidity). According to the results, the effect of irrigation x potassium interaction on the shoot and root sodium (Na) potassium (K) calcium (Ca) and phosphorus (P) content Na/K and Na/K ratio was found to be statistically significant. Shoot and root sodium content decreased with potassium applications under drought conditions (33%). Shoot and root potassium, phosphorus content increased with potassium applications in both drought and sufficient water conditions. Shoot calcium content change irregular with potassium application while root calcium decreased with potassium application under drought conditions (33%). Shoot and root sodium/potassium ratio decreased with potassium applications in both drought and sufficient water conditions. Shoot and root sodium/calcium ratio change irregular with potassium applications. Therefore, it can be said that potassium may play a critical role in reducing the negative effect of drought stress and uptake plant nutrition in sugar beet.

scholarly journals Determination of expression profile of p53 gene in different grades of breast cancer tissues by real time PCR

2020 ◽  
Vol 20 (3) ◽  
pp. 1273-1282
Author(s):  
Haleema Sadia ◽  
Munir Ahmad Bhinder ◽  
Asma Irshad ◽  
Beenish Zahid ◽  
Rais Ahmed ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Gene Expression ◽  
Tumor Suppressor ◽  
Real Time ◽  
Real Time Pcr ◽  
P53 Gene ◽  
Abnormal Behavior ◽  
Internal Stresses ◽  
Down Regulation ◽  
Significant Difference

Background: Pakistan has a high incidence of breast cancer in Asia, where annually 16,232 deaths are reported. There are many exogenous and endogenous risk factors that affect the tumor suppressor genes and oncogenes. The p53 gene is a tumor suppressor gene and it has a role to protect the whole genome from external and internal stresses, which causes damages to the genome. Objective: The aim of the current study was to investigate the p53 gene expression by using the real-time PCR technique in different grades of breast cancer as compared to the normal tissue. Methods: Fresh Modified Radical Mastectomy (MRM) samples (grade1-grade3) were collected from different hospitals of the Lahore. The project was approved by an ethical review committee of Jinnah Hospital, Lahore. And before sampling an informed consent was obtained from patients and clinicians. RNA from fresh biopsies was extracted by Qiagen extraction kit and cDNA was formed. Real time PCR performed by using SYBR green master mix (ABI) and the data was evaluated by using Livak method. Statistical analysis was done by using Microsoft Excel. Results: There was an abnormal gene expression of p53 in all grades of the breast tumors. Non-significant (p>0.05) differ- ence of down and up regulation of p53 in different grades of breast tumor was found. However, as a whole up-regulation was more than down-regulation with significant difference (p<0.0011). Conclusion: The abnormal expression of p53 shows that there are some genetic and epigenetic factors which are the primal cause of an abnormal gene expression. It is recommended that perform next generation sequencing (NGS) of the gene to find out the mutations causing the abnormal behavior of p53 gene. Keywords: Breast cancer; up-regulation; down-regulation; real-time PCR; Punjab; Pakistan; p53 gene.

scholarly journals In Vitro Assessment of the Cell Metabolic Activity, Cytotoxicity, Cell Attachment, and Inflammatory Reaction of Human Oral Fibroblasts on Polyetheretherketone (PEEK) Implant–Abutment

Polymers ◽  
2021 ◽  
Vol 13 (17) ◽  
pp. 2995
Author(s):  
Tzu-Yu Peng ◽  
Yin-Hwa Shih ◽  
Shih-Min Hsia ◽  
Tong-Hong Wang ◽  
Po-Jung Li ◽  
...  
Keyword(s):  
Gene Expression ◽  
Metabolic Activity ◽  
Morphological Changes ◽  
Cell Activity ◽  
Tetragonal Zirconia ◽  
Polymeric Materials ◽  
Cytokine Gene Expression ◽  
Cell Metabolic Activity ◽  
Significant Difference ◽  
Implant Abutment

The purpose of this research is to compare the cytotoxicity of polyetheretherketone (PEEK) and polyetherketoneketone (PEKK) with conventional dental implant–abutment materials, namely titanium alloy (Ti-6Al-4V) and yttria-stabilized tetragonal zirconia polycrystal (Y-TZP), to evaluate the cell metabolic activity, cytotoxicity, and inflammation potential of human oral fibroblasts (HOF) on these materials. Disk-shaped specimens were designed and prepared via a dental computer-aided manufacturing technology system. Surface topography, roughness, and free energy were investigated by atomic force microscope and contact angle analyzer; cell metabolic activity and cytotoxicity by MTT assay; and morphological changes by scanning electron microscopy (SEM). The effect of pro-inflammatory gene expression was evaluated by RT-qPCR. The obtained data were analyzed with one-way analysis of variance and post-hoc Tukey’s honest significant difference tests. PEEK and PEKK exhibited higher submicron surface roughness (0.04 μm) and hydrophobicity (>80°) than the control. Although the cell activity of PEEK was lower than that of Ti-6Al-4V and Y-TZP for the first 24 h (p < 0.05), after 48 h there was no difference (p > 0.05). According to the cell cytotoxicity and the pro-inflammatory cytokine gene expression assays, there was no difference between the materials (p > 0.05). SEM observations indicated that HOF adhered poorly to PEKK but properly to Ti-6Al-4V, Y-TZP, and PEEK. PEEK and PEKK show comparable epithelial biological responses to Ti-6Al-4V and Y-TZP as implant–abutment materials. Between the two polymeric materials, the PEEK surface, where the HOF showed better cell metabolic activity and cytotoxicity, was a more promising implant–abutment material.

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