DREGER A, ADAMCZAK A ,?O?YKOWSKA K S , WIELGUS K
036616 DREGER A, ADAMCZAK A ,?O?YKOWSKA K S , WIELGUS K (Biotechnology Dep, Institute of Natural Fibres and Medicinal Plants, Poland, Email: mariola.dreger@iwnirz.pl) : Pharmacological properties of fireweed (Epilobium angustifolium L.) and bioavailability of ellagitannins.. Herba Polo 2020, 66(1), 1-13.
Fireweed (Epilobium angustifolium L.) is a well-known medicinal plant traditionally used in the treatment of urogenital diseases, stomach and liver disorders, skin problems, etc. E. angustifolium extracts show antiandrogenic, antiproliferative, cytotoxic, antioxidant, anti-inflammatory, immunomodulatory, and antimicrobial activities. The unique combination of biological properties demonstrated by the results of some studies indicates that fireweed has a positive effect in benign prostatic hyperplasia (BPH) and potentially in the prostate cancer chemoprevention. However, the efficacy of E. angustifolium phytotherapy is still poorly tested in clinical trials, while numerous beneficial effects of extracts have been documented in the in vitro and in vivo tests. Fireweed is rich in polyphenolic compounds, particularly ellagitannins. Currently, polyphenols are considered to be modulators of beneficial gut microbiota. The literature data support the use of ellagitannins in the prostate cancer chemoprevention, but caution is advised due to the highly variable production of urolithins by the individual microbiota. A better understanding of the microbiota’s role and the mechanisms of its action are crucial for an optimal therapeutic effect. This paper aims to summarize and discuss experimental data concerning pharmacological properties of E. angustifolium and bioavailability of ellagitannins – important bioactive compounds of this plant.
1 illus, 1 tables, 74 ref
KAPILDEV G, CHINNATHAMBI A, SIVANANDHAM G, RAJESH M, JEYARAJ M, SELVARAJ N, ALHARBI S A, GANAPATHI A
036535 KAPILDEV G, CHINNATHAMBI A, SIVANANDHAM G, RAJESH M, JEYARAJ M, SELVARAJ N, ALHARBI S A, GANAPATHI A (Microbial Biotechnology Dep, Bharathiar Univ, Coimbatore-641 046, Email: aganapathi2003@rediffmail.com) : Meta-Topolin and ?-cyclodextrin enhance multiple shoot and root production in black gram Vigna mungo (L.) hepper. Indian J Expl Biol 2020, 58(5), 314-22.
The recalcitrant nature of black gram is the major constraint of in vitro regeneration and agrobacterium- mediated genetic transformation, to overcome this, a productive shoot regeneration protocol has been achieved in black gram cultivar T9 using 7-day old cotyledonary node explants excised from in vitro- raised black gram seedlings using meta-topolin. An aromatic cytokinin, meta-topolin along with BA (1.5+0.5 mg/L) in 0.50 strength MS medium with 1.5 % (w/v) sucrose exhibited a maximum number of multiple shoots (32.0±0.37 shoots/explant) at the end of 6 weeks of culture. The shoots were elongated in (6.40±0.50 cm/shoot) in MS medium supplemented with GA3 (2.0 mg/L). A maximum number of roots (9.60±0.50/shoot) and root length (11.20±0.73 cm/shoot) were obtained in combination with β-cyclodextrin (a cyclic oligosaccharide; 1.5 mg/L) and IBA (1.5 mg/L). The rooted plantlets were hardened and acclimatized with least mortality rate of 2 % in pot mixture consisting red soil:sand:farm yard manure (FYM) (2:1:1) and grown in green house with 85 % relative humidity. Ploidy levels were analyzed using flow cytometry which confessed the chromosomal stability in invitro raised plants similar to parent plants. This protocol may be useful for producing transgenic black gram with desirable agro-traits in Indian cultivars.
4 illus, 5 tables, 26 ref
TIPPABATHANI J, NELLORE J, KATHIRKANNAN P, NACHIYAR C V
036523 TIPPABATHANI J, NELLORE J, KATHIRKANNAN P, NACHIYAR C V (Biotechnology Dep, Sathyabama Institute of Science and Technology, Chennai, Tamil Nadu, Email: sree_nellore@yahoo.com) : Developmental effects of three textile chemicals on locomotor activity, antioxidant markers and acetylcholine esterase activity in zebrafish. Indian J Expl Biol 2020, 58(3), 212-8.
Textile chemicals discharged into the water bodies cause huge impact on human health and environment. However, the adverse effects of textile chemicals during critical period of brain development are not explored. This study uses zebrafish to assess the developmental toxicity of three textile chemicals. Zebrafish embryos were exposed to different concentrations (1, 5, 10, 20 and 40 PPM) of Naphthalene sulfonic acid (NSA), Metanilic acid (MA) and Acid blue 113 (AB113) from 18 h post-fertilization (HPF) to 96 HPF, respectively. Several endpoints, such as mortality, morphological abnormalities and locomotor activity of embryos and larvae were studied. Biochemical detection of oxidative stress, glutathione and acetylcholine esterase was subsequently tested. The survival rate was decreased (LC (50): 1PPM) by NSA, MSA or AB113 and at > 5PPM a90 % mortality was observed respectively. Exposure to 1 PPM of NSA, MSA or AB113 significantly reduced the locomotoractivity in an age dependent manner. However, no neurodegenerative phenotypes were noted. The glutathione and acetylcholineesterase activity (P <0.05) was decreased while malondialdehyde content was accumulated by NSA, MSA or AB113 treatment. The overall findings suggest that the selected textile dyes exposed during critical window development is able to produce oxidative stress and exert noticeable effects on locomotor activity in zebrafish embryos by altering acetylcholine esterase activity.
6 illus, 36 ref
SIROHI P, YADAV B S, AFZAL S, MANI A, SINGH N K
035984 SIROHI P, YADAV B S, AFZAL S, MANI A, SINGH N K (Biotechnology Dep, Motilal Nehru National Institute of Technology Allahabad, Prayagraj - 211 004, Email: singhnand@gmail.com) : Identification of drought stress-responsive genes in rice (Oryza sativa) by meta-analysis of microarray data. J Genet 2020, 99, 35.
Meta-analysis provides a systematic access to the previously studied microarray datasets that can recognize several common signatures of stresses. Three different datasets of abiotic stresses on rice were used for meta-analysis. These microarray datasets were normalized to regulate data for technical variation, as opposed to biological differences between the samples. A t-test was performed to recognize the differentially-expressed genes (DEGs) between stressed and normal samples. Gene ontology enrichment analysis revealed the functional distribution of DEGs in different stressed conditions. Further analysis was carried out using software RICE NET DB and divided into three different categories: biological process (homoiothermy and protein amino acid phosphorylation), cellular component (nucleus and membrane), and molecular function (zinc ion binding ad DNA binding). The study revealed that 5686 genes were constantly expressed differentially in Oryza sativa (2089 upregulated and 3597 downregulated). The lowest P value (P = 0.003756) among upregulated DEGs was observed for naringenin, 2-oxoglutrate 3-dioxygenase protein. The lowest P value (P = 0.002866816) among the downregulated DEGs was also recorded for retrotransposon protein. The network constructed from 48 genes revealed 10 hub genes that are connected with topological genes. These hub genes are stress responsive genes that may also be regarded as the marker genes for drought stress response. Our study reported a new set of hub genes (reference genes) that have potentially significant role in development of stress tolerant rice.
7 illus, 5 table, 33 ref
SIDDIQUI M Z, AHMAD G, AMIN K M Y, AKHTAR S, REHMAN A
035840 SIDDIQUI M Z, AHMAD G, AMIN K M Y, AKHTAR S, REHMAN A (Biotechnology Dep, Faculty of Natural Sciences, New Delhi- 110 025, Email: drzakiramu10@gmail.com) : HPLC profiling conclusively distinguished two important Unani drugs, namely, Suranjan Shirin (Colchicum autumnale) and Suranjan Talkh (Colchicum luteum). Indian J Tradl Knowledge 2020, 19, 170-3.
Suranjan (colchicum) is one of the prime drugs used for arthritis in Unani System of Medicine. Two varieties of the drug are available in the market under the name of Suranjan; one is Suranjan Shirin (Colchicum autumnale) and the other is Suranjan Talkh (Colchicum luteum). The two varieties are often confused with each other due to morphological resemblance. So there is a need to set a distinction between these two varieties of Suranjan. For this purpose the marker compound (Total Alkaloid Content) were estimated quantitatively and the High Performance Liquid Chromatography (HPLC) was conducted on both the drugs. 3 g of the powdered drug was extracted in petroleum ether and dissolved in 6 mL of 75 % ethanol to yield test sample. Methanol at a flow rate of 1 mL/min was used as a standard. The peaks eluted were detected at 254 nm and compared with the authentic standard at 3.2 min of retention time. The colchicine concentration was found to be higher in Suranjan Talkh (0.21 %) as compared to the Suranjan Shirin (0.15 %). Therefore the present study offers a phytochemical concentration criterion, namely, colchicine content to distinguish between Suranjan Shirin (Colchicum autumnale) and Suranjan Talkh (Colchicum luteum).
1 illus, 2 tables, 16 ref
AI D, CHENG S, CHANG H, YANG T, WANG G, YU C
025536 AI D, CHENG S, CHANG H, YANG T, WANG G, YU C (China Univ of Mining and Technology, Beijing-100 083, China, Email: caihongyu2013@126.com) : Gene cloning, prokaryotic expression, and biochemical characterization of a soluble Trehalase in Helicoverpa armigera H?bner?(Lepidoptera: Noctuidae). J Insect Sci 2018, 18(3), 1-8.
Trehalase is an indispensable component of insect hemolymph that plays important role in energy metabolism and stress resistance. In this study, we cloned and expressed the gene encoding soluble trehalase (HaTreh-1) of Helicoverpa armigera (cotton bollworm) and characterized the enzyme. HaTreh-1 had a full-length open reading frame encoding a protein of 571 amino acids. Sequence comparison indicated that HaTreh-1 was similar to some known insect trehalases. Two essential active sites (D321 and E519) and three essential residues (R168, R221, and R286) were conserved in HaTreh-1. The recombinant trehalase was expressed in Escherichia coli and purified by nickel exchange chromatography. Molecular weight of the recombinant protein was about 71 kDa, and the optimum HaTreh-1 enzyme activity is at 55 °C with pH 6.0. Enzymatic assays showed a Km value of 72.8 mmol/liter and a Vmax value of 0.608 mmol/(liter·min). Inhibition assays in vitro indicated that castanospermine, a polyhydroxylated alkaloid, was an effective competitive inhibitor of trehalase with a Ki value of 6.7 μmol/liter. The inhibitor action of castanospermine was linked to its modification effect on trehalase structure. The circular dichroism spectrum showed that the percentage of α-helix increased under the presence of castanospermine. Results of our study will aid in developing effective trehalase inhibitors for controlling H. armigera in the future.
7 illus, 2 tables, 64 ref
SUGANTHI M, ARVINTH S, CHANDRASHEKARA K N, RAJKUMAR R, SENTHILKUMAR P
025535 SUGANTHI M, ARVINTH S, CHANDRASHEKARA K N, RAJKUMAR R, SENTHILKUMAR P (Genetic Engineering Dep, SRM Univ, Kattankulathur-603 203, Email: sugenthi88@gmail.com) : Molecular characterization of bacterial biocontrol agents and their chitinase genes from tea soil. Indian J Exp Biol 2018, 56(6), 395-401.
In tea, bacterial biocontrol agents viz. Bacillus and Pseudomonas and an enzyme like chitinase from these bacterial strains are used to control tea pests and pathogens. However, literature on molecular identification of the same is quite scarce. In this study, Bacillus and Pseudomonas strains isolated from tea soil samples, were systematically identified by 16S rRNA sequencing. Molecular characterization of bacteria was carried out to identify the species of different level chitinase producing bacteria and diversity among them. Further, chitinase gene was characterized from these bacteria to understand the gene diversity among different bacterial chitinase that has potential application in controlling the plant pests and pathogens. Sequence analysis of 16S rRNA and chitinase gene sequences was made among thirteen Bacillus and five Pseudomonas species submitted in NCBI Genbank
5 illus, 1 table, 31 ref
VERMA B, THAKUR Y, TRIPATHI M, PARDHI M, KHILARI R, PANDE R
025534 VERMA B, THAKUR Y, TRIPATHI M, PARDHI M, KHILARI R, PANDE R (Pt. Ravishankar Shukla Univ, Raipur- 492 010, Email: rama.pande11@gmail.com) : N-arylhydroxamic acids as a drug like molecule: A motif of binding mode with calf thymus DNA. Indian J Biochem Biophys 2018, 55(3), 215-21.
A drug-like molecule, which has a propensity of binding with DNA play a vital role in drug designing mechanism. In this paper, we tried to find out the DNA binding affinity of two derivatives of N-arylhydroxamic acids: (i) N-p-Chlorophenyl-2- methoxybenzohydroxamic Acid (Cl-2-MBHA) and (ii) N-p-Chlorophenyl-3-methoxybenzohydroxamic Acid (Cl-3-MBHA) with calf thymus DNA (ct-DNA) by applying techniques such as UV-visible spectroscopy, Fluorescence spectroscopy, and Viscometry measurements. The findings concluded with experimental techniques were verified with theoretical calculation using computer-based method, Molecular Docking. Absorption spectra revealed that both the hydroxamic acids derivatives bind to ct-DNA, among two, Cl-2-MBHA exhibits the higher value of binding affinity Kb (9.52 × 103±0.08 M-1). Fluorescence spectra showed that ct-DNA successfully quenches the emission spectra of N-arylhydroxamic acid. Ethidium bromide displacement method was used as a standard for analyzing the mode of binding. Both the hydroxamic acids were found to be groove binders. The Stern–Volmer Constant was found to be 2.05 × 10-2 ± 0.001 M-1 and 3.35 × 10-2 ± 0.002 M-1 for Cl-2-MBHA and Cl-3-MBHA respectively. Theoretical analysis molecular docking was done using Hex software for validating the experimental findings. Hence, it was observed that both experimental and computational method complimented the results and deduces groove binding as the mode of interaction.
9 illus, 1 table, 43 ref
SANKARGANESH D, RAMCHANDRAN R, ASHOK R, SARVANAKUMAR V R, SUKRITA R, ARCHUNAN G, ACHIRAMAN S
025533 SANKARGANESH D, RAMCHANDRAN R, ASHOK R, SARVANAKUMAR V R, SUKRITA R, ARCHUNAN G, ACHIRAMAN S (Animal Science Dep, Bharathidasan Univ, Tiruchirappalli- 620 024, Email: achiramans@gmail.com) : Buck odor production in the cornual gland of the male goat, Capra hircus? validation with histoarchitecture, volatile and proteomic analysis. Indian J Biochem Biophys 2018, 55(3), 183-90.
In many animals, glandular secretions or pheromones that possess biological moieties contain messages encoded by the intrinsic smell. In male goats, the cornual gland (a sebaceous gland), may synthesize and excrete relevant chemical components that are responsible for the ‘buck effect’. To test this, cornual glands from freshly-slaughtered male goats (N=6) were subjected to histoarchitecture analysis, to infer about the structural alignment, to the GC–MS analysis for volatile compounds and to SDS–PAGE for protein profiling followed by MALDI-TOF to characterize specific protein bands. The gland possesses sebum, vacuoles and hair follicles inferring its capability to synthesize and extrude the scent. We found 14 volatiles in GC–MS analysis, in which 1-octadecanol might be a putative pheromone of buck odor. We identified seven different proteins in SDS-PAGE. Two proteins, 28 and 33 kDa, were highly matched with DNA mismatch repair protein and Abietadiene synthase, respectively, as inferred from MALDI-TOF. Conclusively, the volatiles identified in the cornual gland suggest that the structural microelements of the gland may synthesize (sebum and vacuoles) and release the key volatiles through the hair follicles. The volatile(s) thus produced in male goats either solely or synergistically may confer the buck odor.
5 illus, 1 table, 33 ref
TOMY M J, SHARANYA C S, MAHAPATRA D K, SUREH K J, SABU A, HARIDAS M
025531 TOMY M J, SHARANYA C S, MAHAPATRA D K, SUREH K J, SABU A, HARIDAS M (Biotechnology and Microbiology Dep, Kannur Univ, Palayad– 670 661, Email: mharidasm@rediffmail.com) : In vitro assessment of selected benzoic acid derivatives as anti-inflammatory compounds. J Sci Ind Res 2018, 77(6), 330-6.
Inflammation is a major reason of pathophysiology of several diseases, causing a number of disarrays. The present research involved exploring inhibition of soybean 5-lipoxygenase (5-LOX) by various benzoic acid derivatives (eudesmic acid, veratric acid, cumic acid, and syringic acid) and their kinetics studies, comparing with vanillin as a standard reference. Isothermal titration calorimetry (ITC) was performed to determine kinetics of the biomolecular interactions. Further, molecular docking studies were performed using GLIDE module of Schrodinger software to determine the inter-molecular interactions between the ligand and the target. Result underlined that four of the investigated derivatives exhibited potent inhibition of 5-LOX in a competitive manner with IC50 values <60 µM. ITC study demonstrated that all derivatives bind effectively with a single domain of the target. Molecular docking studies revealed that all the four candidates displayed good interaction with the target. As found from Glide scores, veratric acid interacted stronger than other derivatives (GLIDE Score of -10.72 kcal/mol), forming two hydrogen bonds with residues GLN514 and GLN716, quite similar to the standard reference. ITC and GLIDE score analyses showed difference in the binding strength of the derivatives. This study would help in developing novel inhibitors, anti-inflammation as well as understanding mechanism(s) of ligand-enzyme interaction.
3 illus, 3 tables, 10 ref
WAKEKAR R S, JADHAV P V, KALE P B, MOHARIL M P, NANDANWAR R S, MANE S S, DESHMUKH A G, MANJAYA J, DANI R G
025554 WAKEKAR R S, JADHAV P V, KALE P B, MOHARIL M P, NANDANWAR R S, MANE S S, DESHMUKH A G, MANJAYA J, DANI R G (Agricultural Biotechnology Dep, Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Maharashtra - 444 104, Email: jpraveen26@yahoo.co.in) : Pollen dysfunction causes 'Floral Bud Distortion' in Indian soybean (Glycine max). Agric Res 2018, 7(1), 10-24.
Floral bud distortion (FBD) is a peculiar disorder limiting yield of soybean in central India. Investigations were conducted to understand tissue-specific cytological, biochemical and molecular alterations associated with FBD. The conspicuous morphological symptom seen was extended vegetative phase, which remained green even after R7 (beginning maturity) stage of growth and failure to produce pods leading to complete yield loss in affected plants. Reduced numbers of pollen grains as well as pollens with high percent of sterility were recorded in symptomatic plants. Although the stigma was found to be receptive in the symptomatic plants, pollens showed irregular shape and had a thicker exine wall. Carbohydrate (2.21 times), protein (2.38 times) and chlorophyll (2.17 times) contents were found to be significantly higher in symptomatic plants. Tissue-specific differentially expressed TDFs (transcript-derived fragments) were generated through cDNA-RAPD using 40 primers. Seven primers were found polymorphic and size ranged from 160 to 1100 bp. Re-amplified TDFs were sequenced, and they showed partial homology with characterized (ARF, XM_003529058) and uncharacterized (LOC 100815325) transcription factors. This indicates investigating differentially expressed genes are needed to understand the insights of molecular alterations associated with floral bud distortion.
7 illus, 5 tables, 61 ref
KUMARI N, BATRA N G, SHARMA V
025553 KUMARI N, BATRA N G, SHARMA V (Bioscience and Biotechnology Dep, Banasthali Univ, Rajasthan- 304 022, Email: vinaysharma30@yahoo.co.uk) : Photosynthetic performance and drought-induced changes in activity of antioxidative enzymes in different varieties of Vigna radiata. Agric Res 2018, 7(1), 1-9.
In the present study, we analysed the photosynthetic performance in five varieties of Vigna radiata, viz. vars RMG 268, K-851, RMG 492, RMG 975 and Anand using chlorophyll fluorescence parameters. We observed that var. RMG 268 tended to reach highest effective quantum yield of PSII [F/Fm], maximum apparent electron transport rate [ETRmax] and saturating photosynthetically active photon flux density [PPFDsat], followed by var. K-851. Thus as judged by its photosynthetic performance, ecophysiologically var. RMG 268 seems to be better adapted to the semi-arid environment of the state of Rajasthan, India. On the contrary, var. Anand was least adapted to its environment as indicated by lowest ETRmax, PPFDsat and F/Fm values. The activities of certain antioxidant enzymes of Vigna radiata in response to drought were also examined in var. RMG 268 and var. Anand. The increased activities of antioxidant enzymes, presumed to limit cellular damage, were observed in var. RMG 268. Cellular malondialdehyde content signal indicators of lipid peroxidation were much higher in var. Anand compared to var. RMG 268. These data revealed that var. RMG 268 had high resistance to environmental and drought conditions and thus substantiated our results obtained on the basis of plant performance.
5 illus, 2 tables, 36 ref
TSIARA C G, NIKOLOPOULOS G K, DIMOU N L, PANTAVOU K G, BAGOS P G, MENSAH B, TALIAS M, BRALIOU G G, PARASKEVA D, BONOVAS S, ET AL.
025552 TSIARA C G, NIKOLOPOULOS G K, DIMOU N L, PANTAVOU K G, BAGOS P G, MENSAH B, TALIAS M, BRALIOU G G, PARASKEVA D, BONOVAS S, ET AL. (Cyprus Univ, Cyprus, Email: gknikolopoulos@gmail.com) : Interleukin gene polymorphisms and susceptibility to HIV-1 infection: A meta-analysis. J Genet 2018, 97(1), 235-51.
Some subjects are repeatedly exposed to human immunodeficiency virus (HIV), yet they remain uninfected. This suggests the existence of host-resistance mechanisms. The current study synthesizes the evidence regarding the association between interleukin (IL) gene polymorphisms and HIV susceptibility. Medline, Scopus and the Web of Science databases were systematically searched, and a meta-analysis of case–control studies was conducted. Univariate and bivariate methods were used. The literature search identified 42 eligible studies involving 15,727 subjects. Evidence was obtained on eight single-nucleotide polymorphisms (SNPs): IL1A −889 C>T (rs1800587), IL1B +3953/4 C>T (rs1143634), IL4 −589/90 C>T (rs2243250), IL6 −174 G>C (rs1800795), IL10 −592 C>A (rs1800872), IL10−1082 A>G (rs1800896), IL12B −1188 A>C (rs3212227) and IL28B C>T (rs12979860). The IL1B +3953/4 C>T variant appears to increase the risk of HIV acquisition, under the assumption of a recessive genetic model (odds ratio (OR): 4.47, 95 % CI: 2.35–8.52). The AA homozygotes of the IL10 -592 C>A SNP had an increased, marginally nonsignificant, risk (OR: 1.39, 95 % CI: 0.97–2.01). It reached, however, significance in subanalyses (OR: 1.49, 95 % CI: 1.04–2.12). Finally, the well-studied hepatitis C virus (HCV) infection IL28B (rs12979860) CT/TT genotypes were associated with a 27 % decrease in HIV infection risk, especially in populations infected with HCV (OR: 0.73, 95 % CI: 0.57–0.95). Interleukin signalling is perhaps important in HIV infection and some interleukin genetic variants may affect the risk of HIV acquisition. Approaches targeting specific genes and genomewide association studies should be conducted to decipher the effect of these polymorphisms.
4 illus, 3 tables, 124 ref
CALVELLO R, CIANCIULLI A, PANARO M A
025551 CALVELLO R, CIANCIULLI A, PANARO M A (Biosciences, Biotechnologies and Biopharmaceutics Dep, Bari Univ, Bari, Italy, Email: mariaantonietta.panaro@uniba.it) : Unusual structure and splicing pattern of the vertebrate mitochondrial solute carrier SLC25A3 gene. J Genet 2018, 97(1), 225-33.
The DNA sequence corresponding to the second exon of the SLC25A3 gene is duplicated in vertebrates. The second exon codes for the first transmembrane segment and parts of the immediately adjoining intermembrane and mitochondrial matrix segments. The two genomic exon 2 sequences are 84 % similar in zebrafish (slc25a3b gene), 70 % in chicken, 66 % in mouse and 67 % in human. The amino acid identity is 86 % in zebrafish, 77 % in chicken and 70 % in mouse and human. The two copies of exon 2 are separated by an intronic interval. Translation of both exon 2 sequences would alter the reading frame of the downstream sequence, generating a modified aa sequence which would soon be truncated by a stop codon. As a matter of fact the splicing machinery is tuned in such a way that in some species only one of the two copies is expressed and the other is spliced out, while in other species both copies are expressed but only one at a time, generating two alternative protein products.
5 illus, 1 table, 20 ref
ZHANG J, CHEN J, MA T, GUO H, YANG B
025550 ZHANG J, CHEN J, MA T, GUO H, YANG B (Gastroenterology Dep, Jiangsu Taizhou People’s Hospital, Taizhou, People’s Republic of China, Email: yangbintaizhou@126.com) : Genetic variants of FOXP1 and FOXF1 are associated with the susceptibility of Oesophageal adenocarcinoma in Chinese population. J Genet 2018, 97(1), 213-8.
This study aimed to investigate whether the genetic variants of CRTC1, BARX1, FOXP1 and FOXF1 are associated with the development of oesophageal adenocarcinoma (OA) in Chinese population. A total of 744 OA patients and 1138 controls were included in this study. Here we genotyped four SNPs, rs10419226 of CRTC1, rs11789015 of BARX1, rs2687201 of FOXP1 and rs3111601 of FOXF1. The chi-square test was used to compare the genotype and allele frequencies between the patients and controls. The student’s t-test was used to compare FOXP1 expression in the tumour and the adjacent normal tissues. The relationship between genotypes of rs2687201 and FOXP1 expression was investigated by one-way analysis of variance test. Patients were found to have significantly higher frequency of allele A of rs2687201 and allele C of rs3111601 when compared with the controls (49.2 vs 43.4 %, P = 0.0008 for rs2687201; 29.1 vs 24.0 %, P = 0.0003 for rs3111601). There was a significantly higher expression level of FOXP1 in the tumour than in the adjacent normal tissue (0.0052 ± 0.0021 vs 0.0027 ± 0.0018, P < 0.001). Patients with genotype AA were found to have remarkably higher FOXP1 expression in the tumour than those with genotype CC (P = 0.01). To conclude, the varients of FOXP1 and FOXF1 genes are functionally associated with OA in Chinese population. With the identification of more susceptible loci, the combined effect of these markers may be helpful for the surveillance of OA.
1 illus, 3 tables, 28 ref
ALI S G, DARVISHZADEH R, EBRAHIMI A, BIHAMTA M R
025549 ALI S G, DARVISHZADEH R, EBRAHIMI A, BIHAMTA M R (Plant Breeding and Biotechnology Dep, Urmia Univ, Iran, Email: r.darvishzadeh@urmia.ac.ir) : Identification of SSR and retrotransposon-based molecular markers linked to morphological characters in oily sunflower (Helianthus annuus L.) under natural and water-limited states. J Genet 2018, 97(1), 189-203.
Sunflower is an important source of edible oil. Drought is known as an important factor limiting the growth and productivity of field crops in most parts of the world. Agricultural biotechnology mainly aims at developing crops with higher tolerance to the challenging environmental conditions, such as drought. This study examined a number of morphological characters, along with relative water content (RWC) in 100 inbred sunflower lines. A 10 × 10 simple lattice design with two replications was employed to measure the mentioned parameters under natural and water-limited states during two successive years. In molecular trial, 30 simple sequence repeat (SSR) primer pairs, as well as 14 inter-retrotransposon amplified polymorphism (IRAP) and 14 retrotransposon-microsatellite amplified polymorphism (REMAP) primer combinations were used for DNA fingerprinting of the lines. Most of the examined characters had lower average values under water-limited than natural states. Maximum and minimum reductions were observed in the cases of yield and oil percentage, respectively. The broad-sense heritabilities for all the examined characters were 0.20–0.73 and 0.10–0.34 under natural and water-limited states, respectively. In the studied samples, 8.97% of the 435 possible locus pairs of the SSRs represented significant linkage disequilibrium (LD) levels. In the association analysis using SSR markers, 22 and 21 markers were identified (P ≤ 0.05) for the studied characters under natural and water-limited states, respectively. The corresponding values were 50 and 37 using retrotransposon-based molecular markers. Some detected markers were communal between the characters under water-limited and natural states. This was in line with the phenotypic correlations detected between the characters. Communal markers facilitate the simultaneous selection of several characters and can thus improve the efficacy of selection based on markers in the plant-breeding activities.
5 illus, 4 tables, 53 ref
GOPALAKRISHNAN A, VINEESH N, ISMAIL S, MENON M, AKHILESH K V, JEENA N S, PAULTON M P, VIJAYAGOPAL P
025548 GOPALAKRISHNAN A, VINEESH N, ISMAIL S, MENON M, AKHILESH K V, JEENA N S, PAULTON M P, VIJAYAGOPAL P (ICAR-Central Marine Fisheries Research Institute, Ernakulam - 682 018, Email: vineeshnedumpally@gmail.com) : Mitochondrial signatures revealed panmixia in Lutjanus argentimaculatus (Forsskal 1775). J Genet 2018, 97(1), 179-87.
Mangrove red snapper, Lutjanus argentimaculatus is a commercially important fish. The genetic stock structure of L. argentimaculatus from Indian waters was identified using mitochondrial ATPase 6 and ATPase 8, and cytochrome b (Cytb) genes. A 842 bp region of ATPase 6/8 genes and 1105 bp region of Cytb gene were amplified in 120 samples from six different locations along the Indian coast and obtained 58 and 66 haplotypes, respectively. The high haplotype and low nucleotide diversity values along with mismatch distribution, Tajima’s D and Fu’s Fs analysis suggested a genetic bottleneck events or founder effect, with subsequent population expansion in L. argentimaculatus. Coefficient of genetic differentiation (FST) values was low and nonsignificant for both ATPase 6/8 gene and Cytb genes indicating low genetic differentiation in L. argentimaculatus which can be managed as a unit stock in Indian waters.
5 illus, 1 table, 48 ref
LIANG T, JIA Y, ZHANG R, DU Q, CHANG Z
025547 LIANG T, JIA Y, ZHANG R, DU Q, CHANG Z (Henan Normal Univ, China, Email: 041019@htu.edu.cn) : Identification, molecular characterization and analysis of the expression pattern of soxf subgroup genes the yellow river carp, Cyprinus carpio. J Genet 2018, 97(1), 157-72.
Sox7, Sox17 and Sox18 are the members of the Sry-related high-mobility group box family (SoxF) of transcription factors. SoxF factors regulate endothelial cell fate as well as development and differentiation of blood cells and lymphatic vessels. There is very less information about the functions of these genes in fish. We obtained the full-length cDNA sequence of SoxF genes including Sox7, Sox17 and Sox18 in Cyprinus carpio, where Sox7 and Sox18 had two copies. The construction of a phylogenetic tree showed that these genes were homologous to the genes in other species. Chromosome synteny analysis indicated that the gene order of Sox7 and Sox18 was highly conserved in fish. However, immense change in genomic sequences around Sox17 had taken place. Numerous putative transcription factor binding sites were identified in the 5 flanking regions of SoxF genes which may be involved in the regulation of the nervous system, vascular epidermal differentiation and embryonic development. The expression levels of SoxF genes were highest in gastrula, and was abundantly expressed in the adult brain. We investigated the expression levels of SoxF genes in five specific parts of the brain. The expression levels of Sox7 and Sox18 were highest in the mesencephalon, while the expression level of Sox17 was highest in the epencephalon. In carp, the expression patterns of SoxF genes indicated a potential function of these genes in neurogenesis and in vascular development. These results provide new information for further studies on the potential functions of SoxF genes in carp.
8 illus, 3 tables, 52 ref
ZHENG H, LI H, TAN W, XU C, JIA L, WANG D, LI Z, SUN G, KANG X, YAN F, LIU X
025546 ZHENG H, LI H, TAN W, XU C, JIA L, WANG D, LI Z, SUN G, KANG X, YAN F, LIU X (Henan Agricultural Univ, Zhengzhou 450 002, People’s Republic of China, Email: xjliu2008@hotmail.com) : Oestrogen regulates the expression of cathepsin E-A-like gene through ERβ in liver of chicken (Gallus gallus). J Genet 2018, 97(1), 145-55.
The cathepsin E-A-like, also known as ‘similar to nothepsin’, is a new member of the aspartic protease family, which may take part in processing of egg yolk macromolecules, due to it was identified in the chicken egg-yolk. Previously, studies have suggested that the expression of cathepsin E-A-like increased gradually during sexual maturation of pullets, but the exact regulation mechanism is poorly understood. In this study, to gain insight into the function and regulation mechanism of the gene in egg-laying hen, we cloned the cathepsin E-A-like gene and evaluated its evolutionary origin by using both phylogenetic and syntenic methods. The mode of the gene expression regulation was analysed through stimulating juvenile hens with 17 β-estradiol and chicken embryo hepatocytes with 17 β-estradiol combined with oestrogen receptor antagonists including MPP, ICI 182,780 and tamoxifen. Our results showed that cathepsin E-A-like was an orthologoues gene with nothepsin, which is present in birds but not in mammals. The expression of cathepsin E-A-like significantly increased in a dose-dependent manner after the juvenile hens were treated with 17 β-estradiol (P < 0.05). Compared with the 17 β-estradiol treatment group, the expression of cathepsin E-A-like was not significantly changed when the hepatocytes were treated with 17 β-estradiol combined with MPP (P < 0.05). In contrast, compared with the 17 β-estradiol combined with MPP treatment group, the expression of cathepsin E-A-like was significantly downregulated when the hepatocytes were treated with 17 β-estradiol combined with tamoxifen or ICI 182,780 (P < 0.05). These results demonstrated that cathepsin E-A-like shared the same evolutionary origin with nothepsin. The expression of cathepsin E-A-like was regulated by oestrogen, and the regulative effect was predominantly mediated through ER-β in liver of chicken.
5 illus, 5 tables, 45 ref
QIAN Y, ZHANG Y, WEI B, ZHANG M, YANG J, LENG C, GE Z, XU X, SUN M
025545 QIAN Y, ZHANG Y, WEI B, ZHANG M, YANG J, LENG C, GE Z, XU X, SUN M (Neurology Dep, The Second Affiliated Hospital of Soochow Univ, Suzhou- 215 004, People’s Republic of China, Email: xingshunxu@suda.edu.cn) : A novel Alu-mediated microdeletion in the RUNX2 gene in a Chinese patient with Cleidocranial dysplasia. J Genet 2018, 97(1), 137-43.
Cleidocranial dysplasia (CCD; OMIM: 119600) is a rare autosomal dominant skeletal dysplasia caused by RUNX2 gene mutations. The present study described a sporadic case with CCD. The clinical data of the proband with CCD was reported and genetic analysis was performed. The proband presented with typical CCD features including supernumerary impacted teeth, bilateral clavicle dysplasia, delayed closure of cranial sutures, and short stature; while his hands were normal. Sequencing analysis of the entire coding region of the RUNX2 gene revealed no pathogenic changes; however, copy-number analysis with the Affymetrix HD array found 500 kb genomic microdeletion. Real-time quantitative PCR validated this microdeletion in the 1–4 exons of the RUNX2 gene. The junction point of the breaking DNA was located in the directly oriented AluSz6 and AluSx repetitive elements, indicating that this microdeletion might be generated through an Alu–Alu mediated mechanism. In addition, this microdeletion existed in 21.8 % of the asymptomatic mother’s peripheral blood cells, demonstrating that the mosaicism was not associated with CCD phenotypes. In summary, a pathogenic microdeletion in the RUNX2 gene located on chromosome 6 was responsible for CCD.
3 illus, 23 ref
HE Y, WANG X, WU X, ZHU Y, YANG D
025544 HE Y, WANG X, WU X, ZHU Y, YANG D (Chinese Academy of Fishery Sciences, Hubei- 430 223, People’s Republic of China, Email: yangdg@yfi.ac.cn.) : Expression profiles of amh and foxl2 in Schizothorax kozlovi, and their response to temperature during the early developmental stage. J Genet 2018, 97(1), 127-36.
To elucidate the role of amh and foxl2 in sex differentiation of the teleost fish Schizothorax kozlovi, the full-length cDNAs were cloned from the mature testis and ovary by rapid amplification of cDNA ends (RACE), and their relative mRNA expression levels were determined by quantitative real-time polymerase chain reaction among tissues and temperature groups. The complete amh and foxl2 cDNAs of S. kozlovi were 2060 bp and 1750 bp, which encoded 568 and 306 amino acids, respectively. The amh were expressed only in gonads, while foxl2 was expressed in the gills, brain and gonads, both exhibiting relatively high tissue specificity. The amh exhibited sex-specific expression pattern in the gonads. No sex differences in the foxl2 expression were observed in the brain and gonads, but significant sex differences were found in the gills. No significant differences were found in the foxl2 expression, from the larval to the juvenile stage, and also between different temperature groups. However, significant differences were found in the expression levels of amh from the larval (12–63 days posthatching (dph)) to the juvenile stage (190 dph), and also among the 18C and 10C groups at 31 dph. This result suggested that amh plays an important role in male sex differentiation of S. kozlovi during the early developmental stage, but no similar effect was observed in foxl2.
6 illus, 1 table, 43 ref
MEMON S, WANG L, LI G, LIU X, DENG W, XI D
025543 MEMON S, WANG L, LI G, LIU X, DENG W, XI D (Yunnan Agricultural Univ, China, Email: 2522935343@qq.com) : Isolation and characterization of the major histocompatibility complex DQA1 and DQA2 genes in Gayal (Bos frontalis). J Genet 2018, 97(1), 121-6.
The species origin of Yunnan gayal has been controversial since many years. However, few recent genetic studies have suggested that it has perhaps originated from the hybridization between male Bos frontalis and female B. taurus or B. indicus. Being an important semi-wild bovid species, this has also been listed under the red list of International Union of Conservation of Nature and Natural Resources. However, there is limited information available about the immunogenicity of this precarious species of Bos. Major histocompatibility complex (MHC) plays a pivotal role in immune response to infectious diseases in vertebrates. In the present study, we have investigated the structural and functional characteristics and possible duplication of the MHC-DQA genes in gayal (B. frontalis). Two full-length cDNA clones of the MHC-DQA genes were amplified and designated as Bofr-DQA1 (DQA*0101) and Bofr-DQA2 (DQA*2001) with GenBank accession numbers KT318732 and KT318733, respectively. A comparison between BofrDQA1, Bofr-DQA2 and to other MHC-DQA molecules from different animal species showed that nucleotide and encoded amino acid sequences of these two identified MHC-DQA genes have more similarity to alleles of specific DQA1 and DQA2 molecules from other Ruminantia species than to each other. The phylogenic investigation also demonstrated a large genetic distance between these two genes than to homologous from the other species. The large genetic distance between Bofr-DQA1 and Bofr-DQA2, and the presence of different bovine DQA putative motifs clarify that these sequences are nonallelic type. These results could suggest that duplication of the DQA genes has also occurred in gayal. The findings of the present study have strengthened our understanding to MHC diversity in rare ruminants and mutation of immunological functions, selective and evolutionary forces that affect MHC variation within and between species.
2 illus, 1 table, 36 ref
DUBEY N K, MISHRA D K, IDRIS A, NIGAM D, SINGH P K, SAWANT S V
025542 DUBEY N K, MISHRA D K, IDRIS A, NIGAM D, SINGH P K, SAWANT S V (CSIR-National Botanical Research Institute, Lucknow - 226 001, Email: samirsawant@nbri.res.in) : Whitefly and aphid inducible promoters of Arabidopsis thaliana L.. J Genet 2018, 97(1), 109-19.
Lack of regulated expression and tissue specificity are the major drawbacks of plant and virus-derived constitutive promoters. A precise tissue or site-specific expression, facilitate regulated expression of proteins at the targeted time and site. Publically available microarray data on whitefly and aphid infested Arabidopsis thaliana L. was used to identify whitefly and aphid-inducible genes. The qRT-PCR further validated the inducible behaviour of these genes under artificial infestation. Promoter sequences of genes were retrieved from the Arabidopsis Information Resources database with their corresponding 5UTR and cloned from the A. thaliana genome. Promoter reporter transcriptional fusions were developed with the beta-glucuronidase (GUS) gusA gene in a binary expression vector to validate the inducible behaviour of these promoters in eight independent transgenic Nicotiana tabaccum lines. Histochemical analysis of the reporter gene in T2 transgenic tobacco lines confirmed promoter driven expression at the sites of aphid and whitefly infestation. The qRT-PCR and GUS expression analysis of transgenic lines revealed that abscisic acid largely influenced the expression of both aphid and whitefly inducible promoters. Further, whitefly-specific promoter respond to salicylic acid and jasmonic acid (JA), whereas aphid-specific promoters to JA and 1-aminocyclopropane carboxylic acid. The response of promoters to phytohormones correlated to the presence of corresponding conserved cis-regulatory elements.
6 illus, 60 ref
PAUL P, MALAKAR A K, CHAKRABORTY S
025541 PAUL P, MALAKAR A K, CHAKRABORTY S (Biotechnology Dep, Assam Univ, Silchar - 788 011, Email: supriyoch2008@gmail.com) : Codon usage vis-a-vis start and stop codon context analysis of three dicot species. J Genet 2018, 97(1), 97-107.
To understand the variation in genomic composition and its effect on codon usage, we performed the comparative analysis of codon usage and nucleotide usage in the genes of three dicots, Glycine max, Arabidopsis thaliana and Medicago truncatula. The dicot genes were found to be A/T rich and have predominantly A-ending and/or T-ending codons. GC3s directly mimic the usage pattern of global GC content. Relative synonymous codon usage analysis suggests that the high usage frequency of A/T over G/C mononucleotide containing codons in AT-rich dicot genome is due to compositional constraint as a factor of codon usage bias. Odds ratio analysis identified the dinucleotides TpG, TpC, GpA, CpA and CpT as over-represented, where, CpG and TpA as under-represented dinucleotides. The results of (NcExp−NcObs)/NcExp plot suggests that selection pressure other than mutation played a significant role in influencing the pattern of codon usage in these dicots. PR2 analysis revealed the significant role of selection pressure on codon usage. Analysis of varience on codon usage at start and stop site showed variation in codon selection in these sites. This study provides evidence that the dicot genes were subjected to compositional selection pressure.
5 illus, 3 tables, 55 ref
LI J, ZHU J-L, LOU S-D, WANG P, ZHANG Y-S, WANG L, YIN R-C, ZHANG P-P
025540 LI J, ZHU J-L, LOU S-D, WANG P, ZHANG Y-S, WANG L, YIN R-C, ZHANG P-P (Anhui Univ, Anhui, China, Email: eveyin@163.com) : The complete mitochondrial genome of Coptotermes ?suzhouensis? (syn. Coptotermes formosanus) (Isoptera: Rhinotermitidae) and molecular phylogeny analysis. J Insect Sci 2018, 18(2), 26.
Coptotermes suzhouensis (Isoptera: Rhinotermitidae) is a significant subterranean termite pest of wooden structures and is widely distributed in southeastern China. The complete mitochondrial DNA sequence of C. suzhouensis was analyzed in this study. The mitogenome was a circular molecule of 15,764 bp in length, which contained 13 protein-coding genes (PCGs), 22 transfer RNA genes, two ribosomal RNA genes, and an A+T-rich region with a gene arrangement typical of Isoptera mitogenomes. All PCGs were initiated by ATN codons and terminated by complete termination codons (TAA), except COX2, ND5, and Cytb, which ended with an incomplete termination codon T. All tRNAs displayed a typical clover-leaf structure, except for tRNASer(AGN), which did not contain the stemloop structure in the DHU arm. The A+T content (69.23 %) of the A+T-rich region (949 bp) was higher than that of the entire mitogenome (65.60 %), and two different sets of repeat units (A+B) were distributed in this region. Comparison of complete mitogenome sequences with those of Coptotermes formosanus indicated that the two taxa have very high genetic similarity. Forty-one representative termite species were used to construct phylogenetic trees by maximum likelihood, maximum parsimony, and Bayesian inference methods. The phylogenetic analyses also strongly supported (BPP, MLBP, and MPBP = 100 %) that all C. suzhouensis and C. formosanus samples gathered into one clade with genetic distances between 0.000 and 0.002. This study provides molecular evidence for a more robust phylogenetic position of C. suzhouensis and inferrs that C. suzhouensis was the synonymy of C. formosanus.
3 illus, 6 tables, 68 ref
AL-GHUFAILI M K F, AL-TAMIMI A J T
025539 AL-GHUFAILI M K F, AL-TAMIMI A J T (Kufa Univ, Iraq, Email: atyaf.altameemi@uokufa.edu.iq) : Genetic relationship among ten wheat genotypes using seventeen RAPD markers. Plant Arch 2018, 18(1), 595-600.
This study was conducted for determination DNA fingerprint and estimation of genetic diversity among ten wheat (Triticum aestivum L.) genotypes using seventeen RAPD Markers. Primers OPB-06, OPC-05, OPH-01 and UBC-126 gave unique fingerprint for studied genotypes. Primer OPG-09 gave higher value for polymorphism . The higher efficiency and discriminatory value was produced by primer UBC-126. High genetic distance was 0.546 between Buhuth22 and Faris while low genetic distance was 0.142 between Rasheed and Iraq. Cluster analysis (Phylogenetic tree) by UPGMA based dendrogram revealed that studied genotypes grouped in two main clusters. Low polmorphisim (36.827) revealed by studied primers. Results show that RAPD markers could efficiently reveal genetic variation and fingerprint wheat genotypes .
5 illus, 4 tables, 38 ref
JWAD R A, RUBAIEE H M A, KHALIL F A A, KHALEEL A I
025538 JWAD R A, RUBAIEE H M A, KHALIL F A A, KHALEEL A I (Plant Protection Dep, Kufa Univ, Iraq, Email: rashamohammed2011@gmail.com) : Genetic diversity assessment of some stored insect species in Iraq based on RAPD molecular marker. Plant Arch 2018, 18(1), 546-50.
Genotyping of four insect species was carried out using RAPD marker. The genetic variability among the four insect species was estimated using RAPD primers. All primers generated reproducible and easily storable RAPD profiles with some amplified DNA fragments ranging from 9 to 13. The total number of amplicons detected was 56, including unique bands, reached seven, Number of polymorphism was 49 and this represents a level of polymorphism of 87.04 % and an average number of 6.8 polymorphic bands per primer. Maximum numbers of amplicons were amplified by primer U-17 reached 13 while the minimum number of fragments was amplified with primers OPE K-02 reached nine. The highest number of polymorphic bands reached 12 were obtained with primer U-17, while the highest number of monomorphic bands reached two was obtained with primer OPE K-02, and M-32 with percentage reached 22.22 % and 16.664, respectively. RAPD markers detected genetic similarity and distance, a maximum genetic distance value was observed between T. granarium(1) and C. maculatus(4) and S. oryzae(2) and C. maculatus(4) species reached 0.547 with less similarity value reached 46 %, a minimum genetic distance value was 0.435, which observed between S. oryzae (2) and T. castaneum (3) with high similarity value reached 57 %. The similarity matrices were employed in the cluster analysis to generate a dendrogram using the UPGMA method. The cluster tree analysis showed that the insect species were broadly divided into two main groups A and B with genetic similarity reached 30 %. A group including T. granarium (1) B group was divided into two sub-cluster B1, and B2 with genetic similarity reached 36 %. The first sub-cluster (B1) including only C. maculatus (4) while the second sub-cluster (B2) included two species S. oryzae (2) and T. castaneum (3) with the high genetic similarity between them reached 48 %.
3 illus, 2 tables, 17 ref
ABFULHUSSEIN F R, MUTLG N H, SARHEED A F
025537 ABFULHUSSEIN F R, MUTLG N H, SARHEED A F (Field Crops Dep, Al-Muthanna Univ, Iraq) : Genotypic characterization and tissue localization of the mutant lines expression of OsHKT1;3 gene in rice under salt stress. Plant Arch 2018, 18(1), 489-95.
In rice (Oryza sativa ‘Nipponbare’), the genes family HKT is composed of eight members. It encodes for Na+ transporters that play an important role in salt stress tolerance. Functional analysis, it has been determined that there is a wide diversity of these transporters in terms of their Na+/K+ selectivity and K+ affinity, in addition, plays an important role in reducing Na+ accumulation in shoots to cope with salt stress. This study focuses on one of these genes, yet to be characterized in literature: OsHKT1;3 (also known as HKT6). Using reverse genetics, the aim of this study is to determine the role of this gene during salt stress. In order to observe the expression of this gene in different organs of the plant, GUS transcriptional fusions were made, with GUS as the reporter gene. Then, using the CRISPRCAS9 method, previously, loss of function mutant lines were generated. Physiological test were made on effect of loss of function of OsHKT1;3 on Na+ and K+ accumulation in rice tissues during salt stress on different tissues (roots, sheath and leaf blade). The results obtained show that all mutants are homozygous, induce a frame shift and being present at the beginning of the gene (1st exon). OsHKT1;3::GUS showed strong GUS activity, expressed mainly in vascular tissues and did not show a significant difference in the expression with the NaCland water-treated parts. In contrast, the GUS activity of the OsHKT1;3 promoters in NaCl-treated leaves was greater than that in water-treated leaves. The results of this study show that in wild plants, increasing the Na+ concentration in the culture medium has the effect of increasing the Na+ content of the tissues generally, the old leaves accumulating more Na+ . When the Na+ concentration increases, the K+ content decreases in roots and old leaves, but varies little in young leaves. Analysis of Na+ storage in tissues also shows that Na+ levels are higher in the lower parts of the leaf than in the upper parts. Analysis of Na+ storage in tissues also shows a Na+ in leaf blade more evenly distributed among the first leaves than in WT plants. Taken together, these results suggest that OsHKT1;3 gene plays a role in the accumulation of Na+ in old leaves.
6 illus, 16 ref
HARIBALAGANESH R, ROSY J C, ROHITA S, AISHWARYA C, BRINTHA S, RAHUL S, SUNDAR K
025519 HARIBALAGANESH R, ROSY J C, ROHITA S, AISHWARYA C, BRINTHA S, RAHUL S, SUNDAR K (Biotechnology Dep, Kalasalingam Univ, Krishnankoil- 626 126, Email: sundarkr@klu.ac.in) : Antiviral drugs against Ebola: A structure based virtual screening approach. Indian J Biotechnol 2018, 17(1), 176-84.
Repression of highly lethal Ebola virus outbreaks poses a serious public health challenge. Ebola haemorrhagic fever is a highly lethal disease for which there are no effective therapeutic or preventative treatments. Hence, there is a necessity to discover new candidate drugs that could have the capability to contain Ebola. Structure based virtual screening method is a method that could accelerate the drug discovery process. In this study, various known antiviral drugs were used as ligands to screen for their binding ability to Ebola proteins by using protein ligand docking. As the crystal structure of the Ebola proteins were not available, these structures were predicted using PS2 or RABTOR X and the predicted structures were validated using Ramachandran plot. Six of the existing drugs exhibited a better binding affinity to various Ebola proteins; they could be used as lead compounds in developing a better drug for controlling Ebola.
11 illus, 5 tables, 23 ref
LASINA K V, PIRAMANAYAGAM S
025518 LASINA K V, PIRAMANAYAGAM S (Bharathiar Univ, Coimbatore- 641 046, Email: lisina777@gmail.com) : Pharmacophore modelling, 3D-QSAR and docking study of HIV inhibitor. Indian J Biotechnol 2018, 17(1), 160-75.
Human immunodeficiency virus (HIV), the causative virus for acquired immunodeficiency syndrome (AIDS) has become the world’s greatest dispute. Inhibition of nucleocapsid protein domain 7 NCp7 receptors have been sturdily pursued as a promising target for the treatment HIV AIDS. A set of 36 thioesters derivatives has been reported as HIV NCp7 inhibitor was analyzed by employing PHASE method to inspect the structural requirements for diverse analogues to inhibit NCp7 receptors and to obtain a highly predictive model used for designing of novel NCp7 receptors inhibitors. A united study of pharmacophore prediction, atom based 3D-quantitative structure–activity relationship (QSAR) and molecular docking approaches were carried out on pyridinioalkanoyl thiolesters derivatives to understand their structural requisites and binding mode of the best fitted ligand for NCp7 inhibitory activity. Five point pharmacophore hypothesis AADDR (two acceptors, two hydrogen donor, one aromatic ring) yielded a statistically significant 3D-QSAR model with partial least square (PLS) factors 5, regression coefficient value of (R2) = 0.9625, cross validation coefficient value of (Q2) = 0.7775, root mean square error (RMSE) = 0.1358. The core structure of nucleocapsid NCp7 domain is docked with the compounds obtained from ZINC and NCI. Docking study also revealed the binding orientation of active ligands at active residues of NCp7. Using pharmacophore based database search, 3D-QSAR and docking studies, we identified ZINC29569253 as a stable inhibitor. The geometry and type of this pharmacophore model give emphasis to important binding features which will be useful for the design of selective HIV NCp7 inhibitors.
10 illus, 5 tables, 16 ref
PAI S R, UPADHYA V, HEDGE H V, JOSHI R K, KHOLKUTE S D
025517 PAI S R, UPADHYA V, HEDGE H V, JOSHI R K, KHOLKUTE S D (ICMR- National Institute of Traditional Medicine, Belagavi- 590 010, Karnataka, Email: drpaisr@gmail.com) : In vitro rapid multiplication and determination of triterpenoids in callus cultures of Achyranthes aspera Linn. Indian J Biotechnol 2018, 17(1), 151-9.
Achyranthes aspera Linn. was studied for its in vitro rapid multiplication, callus initiation, proliferation and triterpenoid determination. The plant was also evaluated for pretreatment, sterilization and culture initiation during the study. Best results for vigorous cultures were obtained for plants pretreated with 0.2 % (w/v) carbendazime + 3 drops of Tween 20 for 10 min, and double min) solutions. sterilized In using vitro both plantlet sodium regeneration hypochloride in A. (NaOCl) aspera was (4.0 % achieved v/v for from 2 min) axillary and mercuric explants chloride cultured (HgClon Linsmaier 2) (0.1 % w/v & Skoog for 2 (LS) fortified with 6-benzyl amino purine (BAP) alone and/or in combination with Zeatin (Zn), Kinetin (Kn), thidiazuron (TDZ), naphthalene acetic acid (NAA). The combination of BAP (3.00 mg/L) and TDZ (0.10 mg/L) was the best for multiple shoot induction and rapid proliferation. The developed shoots were successfully rooted on medium containing NAA (1.5 mg/L). Rooted plantlets were regenerated and were successfully established in soil with a survival rate of 90 %. The protocol developed for multiplication is rapid and efficient for in vitro propagation of A. aspera. Further, a combination of 2,4-D (3.00 mg/L) and BAP (0.50 mg/L) fortified in LS basal medium, showed significant response with enhanced growth and proliferation of callus. Callus obtained from the 10 combinations were extracted individually using an ultrasonicator, and the extracts were subjected for reverse phase ultra-fast liquid chromatography (RP-UFLC) analysis. Higher amount of betulinic acid (3-hydroxylup- 20(29)-en-(28)-oic acid) and oleanolic acid (3β-hydroxyolean-12-en-28-oic acid) were accumulated on the medium supplemented with 3.00 mg/L of 2, 4-D and 0.50 mg/L of BAP.
3 illus, 4 tables, 41 ref
WANG J, LI X, ZHANG L, ZHANG Y, PENG Y, GUO M, ZHANG R, HUANG J
025516 WANG J, LI X, ZHANG L, ZHANG Y, PENG Y, GUO M, ZHANG R, HUANG J (Minzu Univ of China, Beijing 100 081, People's Republic of China, Email: wangjunli1698@163.com) : Developing micropropagation protocol and analyzing peroxidase activity during morphogenesis in Arisaema decipiens Schott, a medicinal plant. Indian J Biotechnol 2018, 17(1), 145-50.
An efficient micropropagation system for Arisaema decipiens Schott, an ethnic medicinal plant, has been developed. Calluses were induced from petiole explants on Murashige and Skoog (MS) medium supplemented with 0.5 mg l-1 N6-benzyladenine (BA) and 1.0 mg l-1 2,4-dicholorophenoxyacetic acid (2,4-D). The effect of 1.0 mg l-1 2,4-D in combination with 1.0 mg l-1 BA was much more pronounced for the callus proliferation, and fresh weight of callus increased 11.72 times after 5 week of culture. Significantly more protocorm-like bodies (PLBs) could be obtained on calluses exposed to 0.5 mg l-1 indole-3-acetic acid (IAA) in combination with 0.5 mg l-1 BA. In this medium, the PLBs transformed into shoots, and frequency of shoot induction was about 82 %. Up to 100 % of the regenerated shoots formed complete plantlets on MS medium supplemented with 0.5 mg l-1 IAA in combination with 0.5 mg l-1 BA, with an average of 10.35 roots and 7.43 cm long per shoot. Peroxidase (POD) expressions were measured by polyacrylamide gel electrophoresis during shoot induction and differentiation phases. The results showed that POD activity was connected with the different states of shoot morphogenesis.
2 illus, 3 tables, 20 ref
CHAUHAN R S, JHA S K
025515 CHAUHAN R S, JHA S K (Forest Biology and Tree Improvement Dep, Navsari Agricultural Univ, Navsari- 396 450, Gujarat, Email: sumanfort@gmail.com) : Genetic stable mass propagation of Acacia mangium Willd. from mature plus tree. Indian J Biotechnol 2018, 17(1), 128-33.
The study describes an efficient, reproducible and stable in vitro propagation protocol for Acacia mangium from nine years old phenotypically superior plus tree. For effective control of contamination explants were dipped in absolute alcohol for 1 minute followed by 0.1 % mercuric chloride solution for 6 minutes. Maximum establishment (87.8 %) and shoot proliferation was achieved with an average value of 5.7 shoots per explants and shoot length of 2.8 cm in Murashige & Skoog (MS) media supplemented with 6.66 μM 6-benzylaminopurine (BAP) and 0.465 μM kinetin. The addition of 9.80 μM indolebutyric acid (IBA) in half MS media gave maximum rooting (88.3 %) with average 2.7 roots per microshoots with longest root of 18 cm. Rooted plants were hardened and successfully established in the soil. Random amplified polymorphic DNA (RAPD) profile of micropropagated plants shown no change in genetic fidelity up to four growth cycles.
4 illus, 4 tables, 26 ref
SINGH N, MAHAR K S, VERMA S, MEENA B, ROY R K, TEWARI S K, GOEL A K, RANA T S
025514 SINGH N, MAHAR K S, VERMA S, MEENA B, ROY R K, TEWARI S K, GOEL A K, RANA T S (CSIR-National Botanical Research Institute, Lucknow- 226 001, Email: ranatikam@gmail.com) : Molecular analysis of genetic variability and relationship among Gladiolus cultivars. Indian J Biotechnol 2018, 17(1), 118-27.
In the present study, we analyzed genetic variability and relatedness in 62 Gladiolus cultivars using directed amplification of minisatellite DNA (DAMD) and inter simple sequence repeats (ISSR). The maximum inter-cultivar average genetic distance was 0.46 between Tiger Flame and Snow Flower cultivars, while the corresponding least genetic distance 0.14 was between Friendship Pink and Friendship White cultivars, respectively. The cumulative analysis carried out for the data generated with DAMD and ISSR markers showed 83.32 % polymorphism across all the Gladiolus cultivars. This level of polymorphism resulted in the present investigation revealed that the amount of genetic variability in the Gladiolus genome is relatively high. The Jaccard’s similarity coefficient and clustering of genotypes in the unweighted pair group method with arithmetic mean (UPGMA) dendrogram revealed that some of the cultivars are closely related whereas others were found quite distinct from each other. The UPGMA dendrogram resulted in the identification of five major clusters. The present study further demonstrates that DAMD and ISSR are useful markers to elucidate the genetic variability and relationships amongst Gladiolus cultivars, and is a prelude for further utilization of promising and genetically divergent materials in the breeding programmes.
3 illus, 6 tables, 39 ref
GAYACHARAN, BISHT I S, PANDEY A, YADAV M C, SINGH A K, PANDRAVADA S R, RANA J C
025513 GAYACHARAN, BISHT I S, PANDEY A, YADAV M C, SINGH A K, PANDRAVADA S R, RANA J C (ICAR-NBPGR Regional Station, Nainital- 263 132, Email: bishtis@rediffmail.com) : Population structure of some indigenous aromatic rice (Oryza sativa L.) landraces of India. Indian J Biotechnol 2018, 17(1), 110-7.
The Indian sub-continent is home to a large number of indigenous aromatic landraces, which may serve as valuable genetic resources for future quality rice improvement to meet the ever growing demand for quality rice. In the era of high yielding modern varieties there is an urgent need to conserve and mainstream in production systems the valuable aromatic landraces occurring in traditional subsistence farming areas across the country. Understanding the population structure and adaptive variations of landraces is considered useful in this regard. In order to explore the genetic structure and diversity, 256 individuals of 8 aromatic landrace populations from four eco-geographic regions were genotyped using 17 SSR markers in the present study. Sixty-nine alleles were recorded and none of them were found common across all landraces. Twenty- six alleles were found unique to an individual landrace population. Kala Joha from Assam was most diverse in terms of effective number of alleles, expected heterozygosity and Shannon’s information index. Analysis of molecular variance (AMOVA) revealed maximum variation of 39.36 % among populations within groups followed by among groups (30.52 %) and the rest (30.12 %) within populations.
3 illus, 4 tables, 31 ref
SABARA P, VAKHARIA D
025512 SABARA P, VAKHARIA D (Biotechnology Dep, Junagadh Agricultural Univ, Junagadh- 362 001, Email: priteshrhsabara@gmail.com) : Genetic diversity study in papaya (Carica papaya L.) cultivars using RAPD and ISSR markers. Indian J Biotechnol 2018, 17(1), 101-9.
Thirteen papaya cultivars and lines were used to assess genetic diversity through dominant PCR based markers (16 RAPD and 12 ISSR primers). RAPD (random amplified polymorphic DNA) primers gave 58 polymorphic bands out of 126 with 47.19 % polymorphism and ISSR (Inter simple sequence repeats) primers produced 49.80 % polymorphism with 37 polymorphic bands out of 65 bands. Dendrogram showed clear grouping ancestrally related papaya cultivars. Principle coordinate analysis (PCoA) showed congruence with dendrogram pattern while bootstrap values at major nodes in both marker systems as well as in pooled data indicated robustness of cluster pattern in dendrogram construction. Analysis of molecular variance (AMOVA) indicated that highly significant genetic variability was obtained through both marker systems while cultivars showed noticeable variation in case of RAPD markers. Mantel’s test between similarity and cophenetic coefficient matrices in different combination showed poor to very good correlation between matrices. This study indicates relation between papaya cultivars based on their ancestral relationship which can be utilized for conservation of important cultivars and development of future strategies for crop improvement programme.
2 illus, 5 tables, 28 ref
BORA L, SINGH A K, KUMAR A, METWAL M
025511 BORA L, SINGH A K, KUMAR A, METWAL M (Agriculture Coll, Nainital- 263 139, Email: lokeshbora36099@gmail.com) : Morphological and microsatellite marker based polymorphic assessment of genetic diversity and relationship of mango (Mangifera indica L.). Indian J Biotechnol 2018, 17(1), 91-100.
Genetic diversity of 19 genotypes of mango was characterized both by morphological and 20 simple sequence repeat (SSR) markers. Characterization of mango genotypes based on morphological and molecular basis is a better approach for designing breeding projects. On the basis of the present findings it was observed that “Sabri” and “Amrapali” showed dwarf stature, while “Swarna Jahangir” was found to be vigorous in its growth. The unweighted pair group method of arithematic- average (UPGMA) dendrogram based on genetic distance segregated the 19 mango genotypes into two main clusters. The polymorphic information content (PIC) values ranged from 0.38 to 0.81. Jaccard’s similarity coefficient values ranged from 0.15 to 0.79 with polymorphism of 77.5 per cent. Three unique fingerprints were identified in three genotypes which can help in varietal identification. A total of 49 loci (42 polymorphic and 7 monomorphic) were detected with amplified size range of 110 to 359 bp. The maximum numbers of loci (i.e. 5) were detected by the primer MiSHRS-48. Out of 20 SSR primers, 18 were polymorphic. Pusa Surya was found to be the most diverse genotype both morphologically and genetically. The similarity for Pusa Surya was 15 per cent with remaining Indian genotypes. The significant variation exists among the genotypes based on morphological and biochemical characters but with the use of SSR markers, assessment of the genetic diversity can help us to plan a future breeding programme using the diverse parent.
3 illus, 5 tables, 58 ref
ATRI C, SHARMA S, BANGA S S
025510 ATRI C, SHARMA S, BANGA S S (Plant Breeding and Genetics Dep, Punjab Agricultural Univ, Ludhiana- 141 004, Email: chhayaatri@pau.edu) : Genome specific microsatellites in wild crucifers: Cross species/genera transferability. Indian J Biotechnol 2018, 17(1), 80-90.
A high degree of colinearity is now known to exist between closely related species in Brassicaceae, which theoretically allows the exchange of markers between them. Objective of this study was to explore the transferability of simple sequence repeat (SSR) primer pairs identified in crop Brassicas to related wild crucifers. Here, we report and validate transferability of 92, 67 and 105 SSR primer pairs, identified in three diploid Brassica genomes, to Brassica fruticulosa, Erucastrum abyssinicum and Diplotaxis tennuisiliquae with respective amplification of 259, 141 and 291 alleles. One thousand six hundred and thirty three (394 in RL 1359, 451 in PBR 210, 402 in RLC 1 and 386 in UP) alleles were detected in cultivated species. In cultivated Brassica species, the average number of alleles amplified were 3.31, 2.19, 2.97 whereas in wild species the values recorded were 2.70, 1.67, 2.32 for A, B and C genome SSR markers, respectively, however respective average allele values per germplasm set were 2.82 (cultivated) and 2.23 (wild). The average polymorphism information content (PIC) values of A genome primer pairs in wild species was 0.77, ranging from 0.32 to 0.94. These transferable markers can now be exploited for further genetic and introgressive breeding studies. The transferability success generally decreased as the evolutionary distance between the source and target species increased.
3 illus, 2 tables, 27 ref
SINGH A, SHARMA J K
025509 SINGH A, SHARMA J K (Seed Science and Technology Dep, C S K Himachal Pradesh Krishi Vishwavidyalaya, Palampur- 176 062, Email: singhadr@yahoo.com) : Molecular characterization of parental lines and hybrids of maize cultivated in north-western Himalayan region using microsatellite markers. Indian J Biotechnol 2018, 17(1), 74-9.
Maize is an important cereal crop of north-western Himalayan region of India with wide ranging and diversified uses which is resulting in the release and recommendation of new hybrids of maize for the region. The use of parental lines with a narrow genetic base by various private and public sector seed agencies may cause a risk of genetic diversity loss. Molecular characterization of hybrids and parental lines by DNA fingerprinting provides knowledge of the genetic relationship among them thus preventing the risk of increasing uniformity. The technique is helpful for molecular identification and hybrid purity testing. The present study was undertaken to carry out microsatellite marker based DNA fingerprinting of ten maize hybrids and their parental lines for molecular identification and assessment of genetic diversity. Twenty three microsatellite markers were found to be polymorphic. Number of alleles per locus ranged from 2 to 6 with a mean of 3.3. Mean expected heterozygosity and polymorphic information content was 0.58 and 0.52, respectively. Average Jaccard’s similarity coefficient observed was 0.46 revealing an average genetic diversity of 54 % among various genotypes studied. Cluster analysis delineated the maize hybrids and parental lines into four main clusters, A, B, C and D at 0.32 similarity coefficient.
2 illus, 2 tables, 20 ref
KUMAR S, SOHU V S, SINGH R P, GUPTA S K, SRIVASTAVA P, BAINS N S
025508 KUMAR S, SOHU V S, SINGH R P, GUPTA S K, SRIVASTAVA P, BAINS N S (Plant Breeding & Genetics Dep, Punjab Agricultural Univ, Ludhiana- 141 001, Email: kumarsatish227@gmail.com) : Investigating the role of high molecular weight glutenin subunits (HMW-GS) protein in end use quality of Indian flat breads. Indian J Biotechnol 2018, 17(1), 65-73.
Indian unleavened flat breads more commonly known as chapati are core to existence for two-third of its population. Glutenins and gliadins constitute gluten which gives extensibility and elasticity to the dough, traits which are of great importance in bread making. However, limited studies have been attempted to explore the relationship and functionality of the protein subunits with the quality of the unleavened flat breads. Chapati characterization and molecular analysis was carried on two set of genotypes firstly, different commercial wheat cultivars and genetic stocks for various quality parameters, secondly three back cross derived recombinant populations from the parents with different HMW-GS at Glu 1B locus to associate chapati quality with the glutenin subunits. Significant variation obtained in the genotypes revealed that tall varieties are distinct in quality followed by derivatives of C 306 and C 591 (7.8 for both DI 9 and DI 105). The SDS-PAGE analysis of the high molecular weight glutenin protein revealed that the subunit ‘20’ at Glu1B locus is unique to tall traditional wheats. However, such investigations in the recombinant populations indicated complex inheritance of this trait.
2 illus, 5 tables, 15 ref
MYTHILI J B, CHETHANA B S, RAJEEV P R, GANESHAN G
025507 MYTHILI J B, CHETHANA B S, RAJEEV P R, GANESHAN G (Biotechnology and Plant pathology Dep, Indian Institute of Horticultural Research, Bangalore- 560089, Email: jbm@iihr.ernet.in) : Chitinase gene construct from Trichoderma harzianum proved effective against onion purple blotch caused by Alternaria porri. Indian J Biotechnol 2018, 17, 50-6.
Purple blotch caused by Alternaria porri (Ellis) Ciffis the most devastating disease of onion prevalent in different parts of the country. In the absence of a resistant variety in the gene pool, gene transfer technique becomes appropriate as an alternative tool for genetic improvement. Accordingly, chitinase gene especially from the biocontrol agent Trichoderma harzianum Rifai which is used against several fungal pathogens was selected as the candidate gene. Onion transformation is difficult and Allium species are recalcitrant to transformation. Hence, before using this gene for transforming onion for conferring resistance/tolerance to Alternaria porri, chitinase gene was validated against purple blotch pathogen in a model plant system, tobacco. Tobacco transformants with T. harzianum chitinase (Th-chit) gene under the control of a strong constitutive CaMV 35S promoter with NPT-II selection marker were generated. The presence and expression of the transgene was confirmed through PCR and RT-PCR analysis, respectively. Bioassay of primary transgenic plants against A. porri through in vitro detached leaf bioassay revealed significant reduction in lesion size ranging from 73 - 100 % as well as mycelial inhibition to the extent of 25 - 65 % over the control (untransformed) plants. The results suggest that chitinase gene from T. harzianum can be used as a candidate gene for conferring resistance to A. porri in onion.
8 illus, 26 ref
JOSE A A, ANUSREE G, PANDEY A, BINOD P, PANDEY A, BINOD P
025506 JOSE A A, ANUSREE G, PANDEY A, BINOD P, PANDEY A, BINOD P (CSIR-National Institute for Interdisciplinary Science and Technology, Thiruvananthapuram- 695 019, Email: binodkannur@niist.res.in) : Production optimization of poly-γ-glutamic acid by Bacillus amyloliquefaciens under solid-state fermentation using soy hull as substrate. Indian J Biotechnol 2018, 17(1), 44-9.
Poly-γ-glutamic acid (PGA) is a biodegradable polymer with wide applications in the field of medicine, cosmetics, food and agriculture. The aim of the present work was to develop a microbial method to produce PGA under solid-state fermentation. Soy hull was used as the solid media in fermentation by Bacillus amyloliquefaciens. Central composite rotary designs (CCRD) was used to optimize the production of PGA and also studied the pattern of mutual interactions between the variables. The values of P < 0.05 indicated that the model terms were significant with the PGA yield of 111.2 mg/gds at initial moisture 75 %, incubation time 60 h and inoculum size 4.7 ml. Characterization of the purified PGA was done by gel permeation chromatography and fourier transformed infrared (FT-IR) spectroscopy. The choice of soy hull as substrate and the process parameters optimization by response surface methodology (RSM) were found to be significant in PGA production. Due to low-cost nature of the process, this study shows the possibility for establishing large scale production of PGA in cost-effective manner.
4 illus, 4 tables, 22 ref
RATHINASAMY P, THAYUMANAVAN P
025505 RATHINASAMY P, THAYUMANAVAN P (Public Health Dep, Aksum Univ, Ethiopia, Email: sami7bio@gmail.com) : Identification of efficient dye decolorizing laccase producing fungi from Kolli Hills. Indian J Biotechnol 2018, 17(1), 33-43.
Laccase is one of the most promising ligninolytic enzymes for the industrial application and ecofriendly bioremediation process. Twenty-five carpophores were collected from different places of Kolli Hills (Namakkal district Tamilnadu, India) and screened on the solid media containing guaiacol, which enabled the detection of laccase secretion. Three positive strains were isolated and the quantitative production of laccase was determined in submerged culture to select hypersecretory strain for further study. Among the three strains, “ST02” the best producer of laccase was selected and was analyzed for the dye decolorization potential using dyes like Poly R-478 and Remazol Brilliant Blue R (RBBR). Identification of the isolated organism was carried out by classical and molecular methods. Approximately 625 bp of the ST02 5.8S rDNA was amplified by polymerase chain reaction (PCR). The phylogenetic relationship of the isolated strain was studied by comparing the internal transcribed spacer (ITS) sequences of ST02 with similar related sequences deposited in the GenBank database. The present study showed that relatively simple plate test screening method and ITS analysis can be used for identification of laccase producing new strain. The isolated organism was designated as Pleurotus ostreatus IMI 395545.
7 illus, 1 table, 48 ref
VIGNESH V, SATHIYANARAYANAN G, PARTHIBAN K, KUMAR K S, THIRUMUGAN R
025504 VIGNESH V, SATHIYANARAYANAN G, PARTHIBAN K, KUMAR K S, THIRUMUGAN R (Animal Science Dep, Bharathidasan Univ, Tiruchirappalli- 620 024, Email: ramthiru72@gmail.com) : Functional assessment of subtilosin A against Aeromonas spp. causing gastroenteritis and hemorrhagic septicaemia. Indian J Biotechnol 2018, 17(1), 27-32.
Anti-Aeromonas and cell membrane lytic bacteriocin substance, subtilosin A producing Bacillus subtilis VT03 was explored. Strain VT03 was isolated from freshwater fish (Tilapia) intestine and screened for its antimicrobial activity against four pathogenic strains of Aeromonas spp. causing gastroenteritis and hemorrhagic septicaemia. Isolate (VT03) was identified showing inhibition in agar spot assay. The strain VT03 was the one exhibiting strong inhibition and identified as Bacillus subtilis using 16S rRNA sequencing. Cell free supernatant (CFS) of the strain VT03 was active against pathogenic strains of Aeromonas spp, subsequently CFS was partially purified and designated as PPB-VT03 showing inhibition against A. hydrophila ATCC 49140. PPB-VT03 completely lost its activity upon treating with proteinase K revealing that the defense molecule could be proteinaceous in nature. Based on polymerase chain reaction (PCR), functional gene coding for subtilosin A (sboA) was found to be present whereas subtilin (spaS) was absent. The role of partially purified bacteriocin of isolate VT03 (PPB-VT03) through FTIR and SEM analysis revealed the activity of cell lysis. The study demonstrated the potential use of subtilosin A producing Bacillus subtilis as a potent source for antibacterial peptide.
3 illus, 34 ref
BAKHSHI M, EBRAHIMI F, NAZARIAN S, ZARGAN J
025503 BAKHSHI M, EBRAHIMI F, NAZARIAN S, ZARGAN J (Biology Dep, Imam Hossein Univ, Tehran, Iran, Email: kpnazari@ihu.ac.ir) : Computational analysis and gene cloning: Design and preparation of a multi subunit vaccine consisting of EspA, Stx2B and Intimin antigens against enterohaemrrhagic Escherichia coli. Indian J Biotechnol 2018, 17(1), 16-26.
Escherichia coli O157:H7 is an intestinal pathogen that made diarrhoea, haemolytic uremia syndrome (HUS) and hemorrhagic colitis (HC) in patients. Roles of EspA and Intimin at the beginning of bacteria colonization in intestine are critical. Destruction of protein synthesis route with shiga toxins of E. coli O157:H7 is mediated through B-subunit of toxins. In this study, in silico approaches were performed to design a suitable construct from EspA, Intimin and Stx2B and a recombinant chimeric antigen was produced. Bioinformatics analyses such as physicochemical data, mRNA folding, 3D structures of chimera and various immunoinformatic data, such as linear and conformational B-cell epitopes, T-cell epitopes were reported according to authentic data base. The chimeric gene was prepared as synthetic construct after designing and cloning. The validation result showed that 83.9 % residues lie in favoured or additional allowed region of the Ramachandran plot. Epitope prediction results proved very good distribution of conformational B-cell epitopes in the 3D structure of chimera. The identified T-cell epitopes are apt to bind MHC molecules. A good quantity of recombinant chimeric antigen was achieved in host cells. From in silico approach, an appropriate multi subunit vaccine candidate was designed and prepared for immunological examinations.
9 illus, 5 tables, 35 ref
GANESHPURKAR A, SALUJA A
025502 GANESHPURKAR A, SALUJA A (Gujarat Technological Univ, Ahmedabad- 382 424, Email: akspharmacy@yahoo.com) : In silico interaction of rutin with some immunomodulatory targets: A docking analysis. Indian J Biochem Biophys 2018, 55(2), 88-94.
The use of plant products as immunomodulator has a long history. Forms eternal era, plant, mineral and animal products are used as drugs for the treatment of various diseases. The present-day synthetic compounds find their leads in natural products. The process of immunomodulation retunes the immune system of an individual by restoring routine functions. Research on immunomodulators from natural sources has been extensively studied for modulation of immune system along with protection and prevention of diseases. Rutin, a flavonoid has been investigated for its potential immunomodulatory effects. Increased particle clearance, decreased inflammation, increased leucocyte and antibody production was observed with administration of rutin in rats. The present study is focused on exploring in silico interaction of rutin with some chemokines and inflammatory targets. In this study, rutin was docked with TNF-α, IL-1β, IL-6, and NOs. Docking studies revealed the excellent interaction of rutin with these targets. The result of present study provides insight for the discovery of novel molecules for immunomodulation and treatment of inflammatory disorders. Findings from the present study show that rutin may interact with several chemokines and inflammatory mediators. Further studies on rutin and associated flavonoids are necessary to develop and establish QSAR and QSPR studies which may serve a stepping stone for the development of novel and safe immunomodulator. Rutin therefore, can be considered for development of an immunomodulatory agent.
4 illus, 35 ref
WAGH S S, SAHOO N R, RAWAT C, VERMA A D
025532 WAGH S S, SAHOO N R, RAWAT C, VERMA A D (ICAR- Indian Veterinary Research Institute, UP- 243 122, Email: vet.nihar@gmail.com) : Jejunal expression of Mucin4 gene in pigs differentially adhesive to diarrhoeagenic E. coli. Environ Ecol 2018, 36(2), 443-5.
The E.coli mediated piglet diarrhoea is the major cause of the piggery industry which causes severe economic hardship to piggery sector. E. coli binds to the brush border of the epithelial cells of the intestine on this basis of receptors and fimbrae adhesion pattern. Among the putative candidate genes associated with adhesion pattern, mucin4 gene was localized on targeted region of SSC13 and considered as a putative candidate gene due to its biochemical property as well as physical location. The presentinvestigation was conducted to study mucin4 gene expression profile in different adhesion phenotypes. A total of 4 types of adhesion pattern were observed with different frequencies. RT-PCR analysis revealed that mucin4 mRNA expression level was different across the differentially adhesive phenotypes.
1 illus, 1 table, 6 ref
SHARMA M R, KUMARI S, MISHRA M
025530 SHARMA M R, KUMARI S, MISHRA M (ICAR- Central Institute of Subtropical Horticulture, Lucknow- 226 101, Email: manasiibsindia@gmail.com) : Micropropagation of wilt resistant, inter-specific (Psidium molle x Psidium guajava) rootstock of guava. Plant Arch 2018, 18(1), 1170-4.
The present study describes a protocol for in vitro micropropagation of wilt resistant, hybrid rootstock of guava, developed at ICAR- Central Institute for Subtropical Horticulture, Lucknow April to June was found to be the most suitable season for collection of explants. A combination of 0.1 % Carbendazim and 0.05 % Metalaxyl for pre-washing followed by surface sterilization with 0.1 % Mercuric chloride for 7 minutes was found effective in controlling contamination. In order to control oxidative browning, applying wax to the cut ends of the explants yielded better results than the use of antioxidants. Out of all the different media preparations used, MS + 0.1 mg/L IAA+ 4 mg/L BAP+ 30g sucrose gave maximum proliferability (2.6 shoots/explant) and maximum leaf number/shoot (5.6) in the shortest duration viz. 7 days. The regenerated micro-shoots were transferred on rooting media containing MS + 2 mg/L IBA. The rooted plants were acclimatized on sterilized coco-peat supplemented with MS salt solution.
3 illus, 2 tables, 40 ref
AL_AMIRY A H A
025529 AL_AMIRY A H A (Horticulture Dep, Basrah Univ, Iraq, Email: aqeelhadi6@gmail.com) : Comparison between three different methods for isolation genomic DNA from dry date palm leaves without liquid nitrogen. Plant Arch 2018, 18(1), 1011-4.
Three different methods for Isolation genomic DNA from Date palm leaves is described. The leaves was dried and ground before DNA extraction by CTAB development method [100 mM Tris- HCl (pH 8.0), 20 mM EDTA , 4M NaCl, 6 % CTAB, 2 % Polyvinylpyrrolidone, 0.2 % B-mercaptoethanol and 15 µl of proteinase K] and compared with two other methods, the results shows the developing method gives highly quality of DNA (A260/280 = 1.6 to 1.9) in all three Date palm cultivars under tests, this study found the development method is very suitable to DNA extraction from dried leaves without liquid nitrogen.
3 illus, 16 ref
RATHI M H, MOLAN A, ISMAIL M H
025528 RATHI M H, MOLAN A, ISMAIL M H (Biotechnology Dep, Diyala Univ, Diyala, Iraq) : The impact of the extracting solvent and the cultivar on the determination of total phenolic contents and anti-radical activities of extracts from roasted date seeds of two date cultivars cultivated in Iraq. Plant Arch 2018, 18(1), 830-4.
The objectives of the current study were to determine the total phenolic content (TPC) and the anti-radical activities of extracts prepared from roasted date-seeds of two date cultivars (Zahdi and Barhi) using different solvents. The TPC was measured using Folin–Ciocalteau method; while the antioxidant activity was measured using DPPH-radical scavenging assay. The results showed that the extracting had a significant role in the determination of the total amount of the TPC and the free radical scavenging activity of roasted date-seeds. Acidification of the water with hydrochloric acid (1% and 5%) resulted in a significant increase (P< 0.001) in both the TPC and anti-radical activity in comparison with the distilled water alone. Extracts prepared from the powdered roasted seeds of Zahdi cultivar had significantly higher (P< 0.5) TPC and freeradical scavenging activity than their counterparts from Barhi cultivar and in all solvents used. In the extracts from the powdered roasted seeds of both cultivars, positive correlation was found between the TPC and the anti-radical activity, indicating that the phenolic compounds are the main ingredients contributing to the free-radical scavenging activity of the two Iraqi date cultivars. In conclusion, the solvent used for the extraction and the cultivar of the date play a significant role in determining the TPC and free radical scavenging activity and that the roasted date-seeds powder could be a good potential source of bioactive compounds that have anti-radical capacities.
2 illus, 1 table, 26 ref