Research Journal of Biotechnology

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Vol. 15(4) April 2020

Antagonistic activity of lactic acid bacteria isolated from Algerian traditional fermented milks against multi-drug resistant and β-lactamase-producing pathogenic bacteria

Abdelbasset Mechai, Manel Debabza and Souad Zouari

Page No: 1-8

Abstract: The rising problem of the multidrug-resistant strains of food spoilage bacteria and foodborne pathogenic bacteria in food and rapidly decreasing spectrum of antibiotics to control these pathogens has attracted an interest in requiring alternative treatment options. In the present study, a total of 42 isolates of LAB were screened for their inhibitory effect on both gram-positive and gram-negative bacteria involved in various diseases as well as against multidrug resistant bacteria and beta-lactamases producing enteropathogenic bacteria. 10 strains from these LAB were found to produce bacteriocin-like inhibitory substances (BLIS) which have a broad antimicrobial spectrum on tested organisms.

All the produced (BLIS) were fully inactivated by proteolytic enzyme but were unaffected by catalase which indicate their proteinaceous nature. The compounds were heat stable up to 100° C for 20 min and were active over wide range of pH (2 to 10). Due to their selective antibacterial activity, the antimicrobial compounds can effectively be used as biopreservative to inhibit the growth of spoilage bacteria and foodborne pathogens in a wide number of foods.

Evaluation of Antimicrobial activity of Bacterial Endophytes isolated from Tinospora cordifolia

Priti, Battan Bindu and Rani Sulekha

Page No: 9-18

Abstract:Microbial endophytes colonizing in medicinal plants are great reservoir of novel bioactive secondary metabolites that are potential source for pharmaceutical industry. The aim of present investigation was to analyze phytocompounds isolated from bacterial endophytes of Tinospora cordifolia and to further estimate their antimicrobial activity. Four different solvents (ethyl acetate, chloroform, diethyl ether and aqueous) were used to obtain extracts of bacterial endophytes for qualitative phytochemical analysis. The ethyl acetate solvent extract showed the existence of various phytochemicals such as alkaloids, flavonoids and terpenoids which were absent in other solvent extracts (diethyl ether, chloroform and water). Thus, only ethyl acetate extracts of bacterial endophytes were used for determining antibacterial and antifungal activities. Among all endophytic bacterial samples (EB 1-EB39), the samples EB3, EB16, EB20, EB32 and EB35 exhibited best antibacterial activity against E. coli, S. aureus, S. hominis and P. vulgaris when compared to other sample extracts. The samples were identified as B. subtilis strain DSM 10, B. tropicus, B. subtilis strain IAM12118, B. aerius and B. pumilus.

The antifungal activity has also been tested against Alternaria alternata, Helminthosporium nodulosum and Aspergillus niger. The sample extracts i.e. EB3, EB6, EB7, EB16, EB20, EB21 and EB22 revealed good antifungal activity 70.31%, 75.78%, 76.56%, 69.53%, 71.09%, 74.06% and 85.15% against H. nodulosum but endophytic sample EB22 showed maximum inhibition (85.15%) of H. nodulosum growth. Samples EB3, EB13, EB30, EB33, EB35 and EB36 presented maximum inhibition (50%, 49.10%, 52.67%, 67.78%, 46.42% and 67.72%) of A. alternata fungal strain and the endophytic sample EB33 gave best inhibition activity (67.78%) against A. alternata as compared to other sample extracts. The strain Aspergillus niger was not inhibited by any sample extracts. Extracts of sample i.e. EB3 are considered as good antimicrobial agents due to presence of both antibacterial and antifungal activities and are identified as B. subtilis strain DSM10. The results confirmed that Tinospora cordifolia is a rich reservoir of endophytic bacteria which are good source of biologically active compounds responsible for antimicrobial activity like their host plant.

In silico Pharmacokinetic and Molecular Docking analysis of Justicia adathoda compounds targeting Staphylococcus aureusMurB

Josephin Sheeba B. and Selva Mohan T.

Page No: 19-25

Abstract:The current study was focused on the inhibitory activity of alkaloids present in Justicia adhatoda against Staphylococcus aureusMurB protein. The peptidoglycan biosynthetic pathway is one of the best-known processes in these bacteria and the enzymes (MurB) responsible for this pathway are absent in humans. Thus, it was considered as a target for designing of antibiotics compounds. Till date, there is no work undertaken on in-silico analysis of these compounds targeting Staphylococcus aureus MurB protein.

A comparative molecular docking and pharmacokinetic analysis was performed with the standard drug cephalexin vsadathoda compound using different bioinformatics approaches. The results obtained in this study proved that the phytochemicals have better binding affinity than the commercial drug.

Study on biodegradability of PHB produced through different fermentation processes using dextrose as a substrate

Dash Ankita, Mohanty Simanta and Samantaray Devi Prasad

Page No: 26-31

Abstract: PHB is the most common biopolymer that can replace non-biodegradable polymer used in different sectors. Application of PHB in various sectors depends upon its biodegradability, biocompatibility, thermal and mechanical properties regulated by upstream and downstream processing used in fermentation. Here, we studied biodegradation of PHB produced by Bacillus sp. C1 (2013) (KF626477) through different bioprocess technologies. Under submerged fermentation 0.157g/l and 1.09g/l of PHB was produced through sonication di-solvent and sodium hypochlorite digestion multi-solvent extraction, whereas 2.141g/l and 1.967g/l of PHB were recovered through sonication di-solvent and sodium hypochlorite digestion multi-solvent extraction.

Comparatively, higher PHB production was obtained in solid state fermentation than submerged fermentation. However, PHB recovered by sodium hypochlorite digestion multi-solvent extraction under submerged fermentation was degraded faster than others due to its high porosity and rough surface confirmed from SEM imaging. The faster degraded PHB depicted the absorption bands at 1720.43cm-1 corresponding to carbonyl ester group (C=O) of PHB confirmed by FTIR analysis. The high degrees of biodegradability of PHB produced under submerged fermentation indicating various agricultural and biomedical applications.

Phylogenetic identification of Staphylococcus aureus isolated from patients in Ekiti State, Nigeria

Akharaiyi Fred Coolborn and Oyama Marshanty Obem

Page No: 32-39

Abstract:Staphylococcus aureus is an important agent in infection of human diseases and therefore requires adequate identification for information on its effective treatment. Also, most strains of S. aureus species are now found to be resisting the effect of many antibiotics and therefore require urgent attention for alternative drug to combat its disease infection in man. Swabs from human clinical samples (urine, ears, noses and wounds swabs) were streaked onto sterile Petri dishes containing mannitol salt agar, which were incubated at 37 °C for 48 h. Single colonies were purified by streaking method. Agarose gel electrophoresis was used for the extraction of genomic DNA from isolates. Genetic fingerprinting of S. aureus isolated from human clinical samples of patients receiving treatment from three hospitals within Ekiti State was compared using RAPD-PCR markers. Six primers showed good level of polymorphism and reproducibility among thirteen arbitrary primers tested. The amplification reaction of six oligonucleotide primers yielded 135 bands of which 121 were polymorphic with sizes ranging between 200 and 3,000 base pair.

The genetic fingerprinting of the isolated twenty S. aureus species was broadly separated into groups of similarities according to their reaction to antibiotics and plant extracts. The use of phenotypic and molecular methods in the study provided useful information on antibiotic resistance and genetic diversity of S. aureus isolates from patients. The information provided could help in monitoring the evolution of S. aureus strains in Ekiti State and even Nigeria at large over time.

Molecular cloning, characterization and semi-quantitative expression of endochitinase gene from the mycoparasitic isolate of Trichoderma harzianum

Chaudhary Sorabh, Sagar Sushma, Kumar Mukesh, Lal Mehi, Kumar Vinay and Tomar Akash

Page No: 40-56

Abstract:Filamentous fungi from the genus Trichoderma are well known for their biocontrol potential and have been used as antagonistic agents as well as plant growth promoters. Chitinases released by Trichoderma spp. have been capable of hydrolyzing chitin by splitting their β-1, 4-glucosidic bonds. The aim of the present study was to isolate and characterize an endochitinase gene from native Trichoderma harzianum isolate which is involved in mycoparasitism. In total, twelve Trichoderma isolates were screened for chitinolytic activity via dual plate method and greenhouse studies. T. harzianum isolate (SVPRT-THLi03) was selected as a target for isolation, cloning, characterization and expression profiling of an endochitinase gene due to its high chitinolytic activity recorded by the degradation of chitin substrates. The genomic DNA of Trichoderma isolates was amplified and cloned in pGEMT cloning vector.

The recombinant clones were confirmed through colony PCR and restriction analysis. The sequenced 1223 bp clone nucleotide sequence of putative endochitinase gene, ChitTh showed 99% homology to T. harzianum chit-HAR2 endochitinase (AB041752.1) with 0.0 E-value. The complete nucleotide sequence of ChitTh contained a single ORF of 379 amino acids with 40.7 kDa molecular weight and theoretical pI 8.3. The precursor protein contained 22 amino acids long signal peptide at N terminus. Phylogenetic analysis showed that ChitTh protein was clustered into group V with other Trichoderma spp. Semi-quantitative endochitinase gene expression was analysed for different isolates viz. T. harzianum (SVPRT-THLi03 and SVPRT-47) and T. nigricans (SVPPP-7). Among the three isolates, higher expression was observed in SVPRT-THLi03 and SVPRT-47 whereas SVPPP-7 showed lesser gene expression with respect to the other isolates.

Analysis of cis-element distribution and expression of Sucrose Phosphate Synthase, cytoplasmic Phosphogluco mutase and Galacturonosyl transferase genes in rice

Krishna Kumar M., Syamaladevi D.P., Balachandran S.M., Subrahmanyam D., Rajesh K. and Reddy R.R.

Page No: 57-64

Abstract:Sucrose Phosphate Synthase (SPS), cytoplasmic Phosphoglucomutase (cPGM) and Galacturonosyl transferase (GalAT) are among the key genes associated with sucrose and pectin biosynthesis in plants. Analyzing the regulatory cis-elements in the promoter regions of these genes would lead to better understanding on the regulation of gene expression during various developmental stages and environmental conditions. The current study reveals distribution of cis-elements in the promoter regions of rice SPS, cPGM and GalAT through in silico methods and correlates this with corresponding qPCR gene expression pattern. The frequency of vegetative stage related cis-elements was higher in SPS promoter in comparison with cPGM and GalAT.

The observed up- regulation of SPS transcript and down regulation of cPGM and GalAT transcripts during mid-vegetative stage to late-vegetative stage using qPCR supports this prediction. The promoters of these genes were also found to harbour many biotic and abiotic stress responsive cis-elements implicating their role under biotic and abiotic stress conditions.

Morphological and genetic diversity of Colletotrichum gloeosporioides causing mango anthracnose

Archana Sornakili, Raguchander Thiruvengadam and Prabakar Kuppusamy

Page No: 65-73

Abstract: Anthracnose disease caused by C. gloeosporioides is an important pre and post-harvest disease of mango in all mango producing areas of the world. Survey of mango anthracnose in ten different places of Tamil Nadu showed that the level of disease incidence was higher in Neelum variety and the mean per cent disease index was more in Madurai market. Distinctness of C. gloeosporioides in morphological characters among the twenty-six isolates resulted in seven morphological groups and that was correlated with the varieties from which they were obtained.

Among the seven groups derived from this study, the variation was mainly in the colony colour as blackish white, dull white, salmond orange, white, black and the conidia were hyaline with oil globules with the size ranging from 8.29 to 11.52 m x 2.60 to 6.30 m. Diversity of twenty six isolates through RAPD and ISSR markers revealed all the isolates were genetically diversified irrespective of geographical place from where they were collected, their morphological characters and virulent nature of C. gloeosporioides.

Purification and characterization of bacteriocins active against Mycobacterium tuberculosis from lactic acid bacteria

Sivaraj Anbarasu, Vanaja Kumar, Uma Rani, Revathy Kalyanasundram and Manikkam Radhakrishnan

Page No: 74-81

Abstract: The present study was conducted to isolate lactic acid bacteria (LAB) from marine and mangrove sediments of East coast of India, Tamil Nadu and to evaluate their bacteriocins for antimycobacterial properties. Seventeen LAB were isolated from total of 44 marine sediment samples. Eight isolates were found to be Enterococcus sp. in which two isolates viz. LIB04 and LIB06 showed antagonistic activity against M. smegmatis Mc2155.

The potent LAB isolate (LIB04) was identified as Enterococcus hirae by 16s rRNA gene sequencing analysis and named as Enterococcus hirae LIB04. The culture conditions for bacteriocins production were optimal when the medium was maintained at pH 6.5 and without NaCl for 18 hours of incubation. The activity of cell free supernatant (CFS) was completely inactivated after treatment with proteinase K. The enterocin BLIB04 were partially purified with 60% ammonium sulphate precipitation followed by methanol:chloroform extraction. The purified bacteriocin was screened for antimycobacterial activity against M. tuberculosis by luciferase reporter phage (LRP) assay. The results showed that BLIB04 effectively inhibited M. tuberculosis H37Rv, SHRE sensitive and HR resistant M tuberculosis.

Severe Clinical and Neurological manifestations in an Indian Glutaric Acidemia type I patient due to a novel mutation in the Glutaryl-CoA dehydrogenase gene

Muntaj Shaik, Kruthika Vinod T.P., Kamate Mahesh and Vedamurthy A.B.

Page No: 82-85

Abstract:Glutaric-Acidemia type-I(GA-I) is a neurometabolic disease caused by mutations in glutaryl-CoA-dehydrogenase (GCDH) gene. MRI was performed on the patient using the 1.5 T magnet. The patient’s organic acids were measured using dried blood spot (DBS) and a urine sample using MS/MS and GC/MS respectively. DNA was isolated from the patient’s DBS and PCR was performed to amplify all exons and the PCR products were bi-directionally sequenced.

The patient exhibited clinical features and MRI findings compatible with a diagnosis of GA-I. The abnormal elevation of organic acids in the urine supported the presence of glutaryl-CoA dehydrogenase deficiency. Gene sequencing revealed a novel homozygous missense mutation, S190T(g.9896G>C) in exon 6. The present study revealed a novel missense mutation accountable for a severe GA-I phenotype in an Indian patient. This novel mutation will ultimately contribute to a better understanding of the molecular pathology of GA-I disease and shed light on the intricacies of the genotype-phenotype correlation of this disorder.

Nutritional Composition, Antioxidants and Antimicrobial Activities in Muscle Tissues of Mud Crab, Scylla paramamosain

Wan Roslina Wan Yusof, Noorasmin Mokhtar Ahmad, Mohd Alhafiizh Zailani, Mardhiah Mohd Shahabuddin, Ngieng Ngui Sing and Awang Ahmad Sallehin Awang Husaini

Page No: 86-92

Abstract:Mud crab, Scylla paramamosain known as a green mud crab, has become a popular seafood due to its meat quality. In addition, this marine invertebrate has been found to possess peptides with different biological activities and potentials. The aim was, first, to determine the basic nutritional content and second, to screen for the antioxidants and antimicrobials activities in the tissue of mud crab, S. paramamosain. Percentages of carbohydrate, protein and fat in S. paramamosain were 2.32%, 12.53% and 0.23% respectively. The IC50 of methanol extract of tissue S. paramamosain in DPPH scavenging assay was found to be 1.79±0.0 mg/mL. Moreover, superoxide dismutase (SOD) activity was measured as percentage inhibition by SOD was found to be 88.7% ± 2.2, followed by catalase (CAT) activity which was 44.3 ± 2.9 µmol of hydrogen peroxide (H2O2) per minute respectively.

The total glutathione (GSH) in the tissues extract was 0.5 µg/mL. In antimicrobial assay, the methanolic extract of S. paramamosain revealed its potential as antimicrobial agents against three human clinical pathogens with minimal inhibitory concentration (MIC) values ranging between 7 to 62 µg/mL. Minimal bactericidal concentration (MBC) values were higher than 125 µg/mL. The study reported that muscle tissues of S. paramamosain are enriched with high protein content and possess antioxidant and antimicrobial activities. Therefore, further studies are needed to identify and isolate the bioactive compounds in the tissue extracts of S. paramamosain to add more values and potentials of mud crabs.

Efficacy of Chemical Thinning on Plant Growth Parameters in Japanese Plum (Prunus salicina Lindle.) cv. Kala Amritsari

Bhatt Siddharth Shankar, Dimri D.C., Pandey Yamuna and Rawat Devashish S.

Page No: 93-96

Abstract:The plum has been of recent interest for its nutritional, and antioxidant activity. Its consumers have increased considerably. Plum is known to contain considerable vitamins (A, C and E), anthocyanins and other phenolic compounds and carotenoids. Enhanced fruit size is of prime importance to avail such pertinent phytochemicals. Chemical thinning has in recent perspective enhanced fruit size by further enhancing plant growth parameters. Ammonium thiosulphate (ATS @ 1.5%) applied at post bloom stage was most effective in inducing maximum extension of shoot growth (32.45 cm).

The treatment of ATS @ 1.5% applied at post bloom stage showed maximum number of leaves/shoot (33.07) in comparison to other treatments. During the year 2018, size of leaves ranged from 8.68 cm2 (control) to 12.58 cm2 (ATS @ 1.5%). The ammonium thiosulphate treatment (ATS @ 2.0%) when applied at post bloom stage produced foliar toxicity and showed maximum number of damaged leaves (8.20) while under control, minimum phytotoxicity (0.14) T12 (Ethephon @ 200 ppm) was observed. The results showed that ATS 1.5 per cent was best suited to enhance plant growth in plum.

Histopathological evaluation of toxic effects of selenium treated garlic extract on vital organs of healthy Wistar rats

Priyanka, Majumdar R.S., Mishra J. and Deshmukh G.

Page No: 97-106

Abstract:The safety of selenium treated Allium sativum L. (Garlic) is a subject of dilemma. The present study was designed to investigate and compare effects of normally grown and selenium treated garlic leaf aqueous extract (GLAE) on histopathology of myocardium, pulmonary, renal and liver of healthy male Wistar albino rats. Animals were divided in 6 groups and were fed with carrier solution(saline) and GLAE from plants grown in soils(no. 1-5) spiked with increasing concentrations of selenium(Se) (0, 2, 4, 6 and 10 mg/l) as Na2SeO4 at a fixed oral dose of 500 mg/kg body weight (B.W.) for a period of 21 days. Significant toxic effects were observed starting from group 4 (Se concentration 4 mg/l).

There were very minute deflecting effects seen in group 2 and 3 animals from placebo group. No mortality was observed during the complete course of the study. Continual and uncontrolled consumption of garlic plants grown in soil containing > 2 mg/l of Se have gradual and serious health impacts.

Structure-based virtual screening and molecular docking for the identification of potential biofilm inhibitors

Kumar Praveen and Mohanan Archana G.

Page No: 107-115

Abstract:Pseudomonas aeruginosa is an opportunist pathogen that infects the human host when the normal immune defenses are disabled. A biofilm comprises of an aggregation of microorganisms in which cells stick to each other and often also to a surface. These adherent cells become embedded within a slimy extracellular matrix that is composed of extracellular polymeric substances (EPS). WspR, a diguanylate cyclase of P. aeruginosa contains a GGDEF domain known to catalyze the formation of an important biofilm signalling molecule, cyclic diguanylate (c-diGMP). In the present study, the GGDEF domain of WspR was subjected to structure-based virtual screening and molecular docking to identify novel biofilm inhibitors. For virtual screening, iDock, a Web Platform for Large-Scale Protein-Ligand Docking was used.

A set of 243630 compounds from zinc database were screened and 1000 compounds with high docking scores were obtained. The top 10 compounds showing high idock scores were docked again in autodock vina with chimera as an interphase to confirm the binding affinity. Out of 10 compounds, compound 1 (Zinc 84363294), compound 2 (Zinc 77633448) and compound 3 (Zinc 79261305) showed the highest binding affinity to GGDEF domain respectively. Therefore, it is hypothesized that these compounds can be developed as anti-biofilm drugs in the near future. The presented drug discovery based on computational study confirms that several compounds can be used as potential drug candidates against bacterial biofilm and warrants further experimental investigation.

Development of new multiplex nested PCR for rapid detection of Coxiella burnetii

Nemati Afshin Samimi, Zeinoddini Mehdi, Abadi Mohamad Javad Dehghan Esmat and Sheikhi Fatemeh

Page No: 116-120

Abstract:Q fever is a significant worldwide zoonosis caused by Coxiella burnetii, an obligate intracellular, gram-negative and pleomorphic rod. This bacterium caused one type of foodborne diseases and the prevalence of food poisons has increased recently, therefore an accurate diagnosis of this pathogen is necessary. In this study, three sets of specific primers were designed with the goal of multiplex nested PCR detection of C.burnetii. Com1 and IS1111 repetitive elements were chosen for multiplex nested PCR assay. After that, PCR products cloned into the TA vector in order to design a PCR positive control construct.

PCR reaction was performed and three specific bands at 139, 300 and 598 bp were observed in 2% agarose gel electrophoresis stained with KBC power load. The sensitivity of this method was determined about 2.1×10-2 pg for com1 and 12 fg for IS1111. Development of this new PCR reaction is useful for a simple, rapid and correct diagnosis of C.burnetii in food and dairy product. To conclude, this new multiplex nested PCR assay could be used for identification of this important pathogen in research laboratories without culture-based equipment.

Sub-acute Toxicity Assessment of Green Synthesized Hematite Nanoparticles (α-Fe2O3 NPs) using Wistar Rat

Kulkarni Smital, Swain Niharika and Thakur Mansee

Page No: 121-135

Abstract:Toxicological research of nanoparticles is a prime requisite for approval of their clinical use in biomedical field. Since Iron Oxide Nanoparticles (IONPs) such as magnetite nanoparticles (Fe3O4 NPs), hematite nanoparticles (α-Fe2O3 NPs) and maghemite nanoparticles (γ-Fe2O3 NPs) have gained much attention for their potential use in diagnostic and interventional approaches in various diseases, in this study we investigated the safety and biocompatibility of green synthesized hematite nanoparticles (α-Fe2O3 NPs) of size 10-50 nm with spherical shape in Wistar albino female rats. We have analyzed safety of orally administered α-Fe2O3 NPs in three different doses - 1mg/kg, 10mg/kg and 100mg/kg on exposure for 14 and 28 days in rats by assessing various physical, haemotological, biochemical, immunoassay studies and histopathological parameters.

No significant changes were observed in body weight, organ weight, hematological parameters (White blood cell (WBC), Red blood cell (RBC), Haemoglobin (HGB), Hematocrit (HCT), Mean corpuscular volume (MCV), Mean corpuscular hemoglobin (MCH) and Mean corpuscular hemoglobin concentration (MCHC) and Platelet count (PLT)) and immunoassay studies (Serum iron (SI), Total iron binding capacity (TIBC), Serum ferritin (SF) and Transferrin saturation (TS)) in all the groups exposed for 14 and 28 days. In addition to this, biochemical tests such as liver function tests (LFT), renal function tests (RFT), electrolytes, calcium and phosphorous did not show any significant differences in any of the groups exposed for 14 and 28 days. On histopathological examination of organs, majority of them showed signs of toxicity in groups with exposure of 10 mg/kg and 100 mg/kg except brain in which only cellular damage was observed on exposure to highest dose (100 mg/kg). Higher dose and longer exposure of α-Fe2O3 NPs were found to manifest significant signs of tissue toxicity. The present study advocates for need of detailed in vivo study on dose dependent cytotoxic effect of α-Fe2O3 NPs.

Bone morphogenetic protein 2 increasing epithelial to mesenchymal transition in breast cancer cells with pausing of its proliferation

Sharma Tanu, Bandyopadhayaya Shreetama, Dhakar Ramgopal and Mandal Chandi C.

Page No: 136-144

Abstract:Substantial studies pointed the dual role of BMP-2 in tumorigenesis similar to TGF β. Past studies documented that TGF β mostly behaves as a tumor suppressor in case of early stage of a tumor but it often promotes tumorigenic potential in the advanced stage of a tumor. But, the precise role of BMP-2 in tumorigenesis is still opaque. This present study demonstrated that recombinant BMP2 (rh-BMP-2) treatment significantly increased cell migration in breast cancer MDA-MB-231 and MCF-7 cells. It might also augment epithelial to mesenchymal transition (EMT) of breast cancer cells since rh-BMP-2 increased expressions of mesenchymal markers (e.g. vimentin, N-cadherin and Zeb1) with simultaneous decrease of epithelial marker E-cadherin expression. In addition, BMP-2 enhanced expression and activity of matrix metalloproteinases (MMPs).

However, BMP-2 significantly decreased cell proliferation in breast cancer cells with parallel inhibition of cell survival genes BCL2 and Bcl-xL transcripts, and cell cycle markers cyclin D1 and CDK-2 expressions. These findings proposed that BMP-2 might augment metastatic cascade by pausing cell proliferation in breast cancer.

Expression of plant α-amylase inhibitor (αAI) genes revealing bruchid resistance in legume crops

Mishra Sujit Kumar and Panigrahi Jogeswar

Page No: 145-153

Abstract:Legume seeds are the reservoir of nutrition for humans and farm animals. These seeds provide proteins, carbohydrates, lipids, vitamins and minerals in adequate amount. The net productivity of these crops is constantly being affected by an array of destructive pests; the most detrimental pests are the bruchids (Coleoptera: Chrysomelidae). These pests are largely managed by chemical pesticides which are most alarming in the present scenario. These pesticides contain highly toxic chemicals with harmful effects like the undesirable ecological impact on the environment and accumulation of toxic residues in the grain threatening food safety. This insisted for the search of an alternative approach to manage bruchid invasion in legume crops.

Identification of α-amylase inhibitor (αAI) genes in plants and their subsequent exploitation in inhibiting the activity of α-amylases emerged as a promising strategy to manage the bruchid pests in legume crops. With this understanding a number of transgenic legume crops transformed with plant αAI genes have been developed. In this context, this review discusses the expression of plant αAI genes and synchronized bruchid resistance in different legume crops. Further, it highlights the mechanism behind the activity of αAIs against insect α-amylases. The review also illustrates the pros and cons associated with the transgenic legumes containing αAI.

Breeding for herbicide tolerance in crops: a review

Prakash Nitish Ranjan, Chaudhary Jeet Ram, Tripathi Anurag, Joshi Nilesh, Padhan Birendra Kumar, Yadav Suresh, Kumar Santosh and Kumar Rahul

Page No: 154-162

Abstract:Weeds compete with crop plants for water, nutrients, sunlight, and space and also harbor insect and disease pests. With continuously increasing labor cost, manual weeding has become an expensive field operation for any crop and farmers are increasingly opting for cultivars tolerant to herbicides. Herbicide-tolerant cultivars offer opportunity of controlling weeds through need-based applications of herbicides. Herbicide tolerant cultivars have been developed in many crops by exploiting already available genetic variability in the germplasm or by creating mutations or by transgenic. A large genetic variation for tolerance to herbicide exists among various crops such as maize, wheat, rice, sunflower, soybean, chickpea, alfalfa etc. Herbicide tolerance in germplasm or in mutant lines may be due to altered binding site of target enzyme for herbicide, improved herbicide metabolism, sequestration of herbicide molecule and overexpression of target protein.

Various mutations have been reported which may include SNP or insertion/deletion mutation conferring the herbicide tolerance e.g. a point mutation in the AHAS1 gene at C675 to T675 resulting in an amino acid substitution from Ala205 to Val205 confers resistance to IMI in chickpea. Herbicide resistant crop via transgenic has also been developed by taking resistant genes from various sources such as bacteria or other plant and incorporating them into crop to make them herbicide resistant. Transgenic herbicide tolerant crops occupy 47% of total area under transgenic in world. The disadvantage accompanied with herbicide tolerant crops is super weeds. Super weeds are the plants which are crop related weeds species on which the resistance genes got transferred by natural outcrossing.