024548   VERMA S, TOMAR R S, RATHODE V, THAKKER J, SUBHAM, BHAGWAAT N, RAVAL S, ANTALA T, JOGIA Z, GOLAKIYA B A (Biotechnology Dep, SradarkrusinagarAgricultural Univ, S.K Nagar) : Genome sequencing analysis of Macrophomina phaseolina resistant and susceptible castor genotype. Biosci Biotech Res Asia 2018, 15(1), 195-215.

Castor (Ricinus Communis. L, 2n=20) is crop of tropical, sub tropical and warm temperate regions. Castor is most promising crop for the commercial and medicinal values. In recent years castor production is lessen due to Macrophomina phaseolina diseases outbreak which is a necrotrophic soil borne pathogen. Macrophomina phaseolina diseases known as root rot which cause severe diseases especially in dry area. In India it can damage 80-100 % crop. No effective controlling measures are mentioned till date for disease. The more practical practices to control the disease are to identify castor resistant genotype from the Macrophomina phaseolina fungus. There is very less information regarding the molecular aspect of castor JI 357 and 48-1 respectively resistant and susceptible genotype. Thus, resistant genotype of castor provides basis of utilization of molecular approach in the molecular study of resistant genes and mechanism of their gene expression to increase the resistant genotype of castor whereas susceptible genotype helps in the compare the expression and activity of gene. Genome sequencing of resistant and susceptible castor genotype was carried out in Ion Torrent (PGM), Next Generation Sequencer. The data obtained in which resistant sample posses total 2,651,111 reads with total 379,341,629 bases with the average read length of 143 bp with 8 lowest and 597 highest sequence length with 43 % of GC content in resistant genotype and in susceptible total 124,860 reads were generated with total 16,681,708 bases with the average read length of 134 bp with 8 lowest and 572 highest sequence length with 54 % of GC content in susceptible sample of castor. Overall, 401Mb data was generated for resistant and susceptible sample with total reads 2,853,224. Blast2GO tool used to annotate the function of genes according to homologous sequence in resistant castor genotype JI357 out of 100,000 sequences, 89000 sequences were used for GO annotation, 3900 was blasted without hit and 2500 was blast with hit. Among that Ricinus communis have highest hits in resistant genotype while in susceptible 48-1 genotype of castor root in total 18,500 sequence, 4250 sequence used in with GO slim annotation, 9500 sequence used in mapping, 100 sequence found with blast hits and 4500 sequence is with blast (without hits) blast Go analysis.

13 illus, 6 tables, 31 ref

024547   VERMA N M, SAH A K, MAURYA S K (Biotechnology Dep, Invertis Univ, Bareillyy- 243 123, Email: narendra_106@yahoo.com) : Evaluation of G2350A polymorphism of the Angiotensin-converting Enzyme (ACE) gene in chronic kidney disease. Biosci Biotech Res Asia 2018, 15(1), 151-5.

Chronic kidney disease (CKD) becomes a major problem for world health. Numerous studies have documented that the polymorphisms in angiotensin-converting enzyme (ACE) gene may contribute to an individual risk for the loss of kidney function. The present study was undertaken to evaluate the possible relationship between ACE G2350A gene polymorphism and the risk of CKD in Uttar Pradesh population. A total of 379 (159 CKD patients and 220 healthy controls) subjects were recruited for this study. All subjects were genotyped for G2350A polymorphism by PCR-RFLP method. The significant differences were reported between CKD patients and control groups in height, BMI, WC, WH ratio, SBP, DBP, FBS, serum creatinine, eGFR, triglyceride, total cholesterol, HDL and LDL (p < 0.05); while there was no difference in weight, WC, HC and VLDL. The frequency of AA genotype and A-allele were significantly higher in healthy controls than to patients. Conclusively, this study showed that the G2350A polymorphism may not contribute to CKD risk. Further investigations are warranted in larger sample size to confirm our results.

2 tables, 21 ref

024546   ALGHUTHAYMI M A (Biology Dep, Shaqra Univ, Saudi Arabia, Email: malghuthaymi@su.edu.sa) : Cultural, anastomosis and universally primed PCR typing of Rhizoctonia solani from potato. Biosci Biotech Res Asia 2018, 15(1), 119-30.

Forty two isolates of Rhizoctonia solani were recovered from potato in four agro-ecological regions of potato in Saudi Arabia were studied by using anastomosis typing, morphological characters, and molecular method. Almost of isolates sampled from potato cultures belonged to AG-1, AG-3, and AG-7. RP1, RP2, RP3, and RP4 are bridging isolates, they might be classified into another anastomosis groups (AGs) or perhaps might be new anastomosis groups. Universally primed PCR (UP-PCR) fingerprinting was used to evaluate genetic diversity of R. solani AGs infecting potato and other hosts. The majority of the isolates representing various AGs were grouped together into different sub-clusters using all three primers. Molecular groups of the isolates did not correspond to agro-ecological regions or states and crops of the origin. In the current study, the UP-PCR markers could not identify and clearly differentiate the isolates of R. solani. There was no relationship between the geographic origin of the isolates and clustering of isolates based on the genetic diversity. Based on cultural characterization, anastomosis group typing and UP-PCR, no clear-cut conclusions about non-identified anastomosis groups from potato.

8 illus, 2 tables, 30 ref

024545   RAHMAN Z A, ALI M S M, GHAZALLI M N, AWANG K, OTHMAN A N (Biotechnology & Nanotechnology Dep, Research Centre, MARDI HQ, MARDI-UPM- 434 00, Email: azuraida@mardi.gov.my) : Optimization of culture media formulations for micropropagation of Lepisanthes fruticosa. Biosci Biotech Res Asia 2018, 15(1), 51-8.

Tissue culture provides an avenue for the production of high quality clonal plants in large numbers within a short time. Here, we describe the development of protocols for reproducible in vitro micropropagation of Lepisanthes fruticosa via direct organogenesis. Shoots were initiated from two types of explants, nodes and young shoots, to establish in vitro cultures on Murashige and Skoog’s (MS) medium or Woody Plant Medium (WPM) supplemented with different concentrations of benzylaminopurine (BAP). Semi-solid WPM media containing 1 mg/L BAP was most effective in shoot initiation in both node and young shoot explants, giving 40 % and 20 % shoot induction, respectively. The highest rate of shoot proliferation from young shoot explants was obtained using BAP at 3.0 mg/L in combination with NAA at 1.0 mg/L in WPM culture medium. This combination of growth regulators in the medium was also suited to root initiation.

2 illus, 4 tables,17 ref

024544   NANDANPAWAR P C, RATHER M A, BADHE M R, SHARMA R (Fish Genetics and Biotechnology Div, Central Institute of Fisheries Education, Mumbai- 400 061, Email: mashraf38@gmail.com) : Assessment of DNA damage during gene delivery in freshwater prawn by Chitosan reduced gold nanoparticles. Biosci Biotech Res Asia 2018, 15(1), 39-49.

The increasing application of nanoparticles both in industries and in agricultural fields has led to its accumulation in the aquatic ecosystem through water run-off. Insights into the validity of safer nanoparticles such as gold and chitosan are fairly established. However, its effect on aquatic invertebrates has been less studied. The present study was aimed to study effects of chitosan reduced gold nanoparticles (CRGNPs) during green fluorescent protein (GFP) encoding plasmid delivery in giant freshwater prawn, macrobrachium rosenbergii. The mean particle size and zeta potential CRGNPs was 33.7 nm and 24.79 mV respectively. Prawn juveniles were exposed to nanoparticles concentrations (10 μg/L, 20 μg/L) of CRGNPs by immersion treatment for a period of 36 hours. GFP was ubiquitously expressed in muscle tissues of prawns. The comet assay indicated dose dependent genotoxicity of CRGNPs in gill, pleopod and muscle tissues which was in conformity with its bioaccumulation pattern in vivo. The highest bioaccumulation of CRGNPs was found in Gills, followed by pleopods and least in muscles. Hence, the toxicological potential of CRGNPs to the environment cannot be denied and demands more research on the particular aspect. The doses standardized in the present study would be helpful in safer nano-gene delivery in aquatic invertebrates and development of transgenics employing less cost.

7 illus, 61 ref

024543   MAHARANA A K, SINGH S M (National Centre for Antarctic & Ocean Research, Goa- 403 804, Email: drshivmohansinghncaor@gmail.com) : Cold active amylases producing psychrotolerants isolated from Nella Lake, Antarctica. Biosci Biotech Res Asia 2018, 15(1), 5 -16.

Cold active amylase was investigated by bacteria and yeast isolates from the sediment core samples of Nella Lake, Larsemann Hills region, East Antarctica. Between potential yeast and bacteria isolates screened for amylases, best isolates were identified as Rhodotorula sp. Y-37 and Arthrobacteralpinus N16 by molecular technique. Amylase production capabilities of both the isolate subjected for optimization processes by using submerged fermentation technique with soluble starch as substrate. The results indicate that a supplement of 1 % w/v glucose, 1 % w/v yeast extract and 0.1 % w/v KCl at pH 7.0 with 5 % v/v inoculum enhances the amylase production by 5.72-fold using Rhodotorula sp. Y-37. In other hands, the activators are 1 % w/v of galactose and peptone, 0.1 % w/v KCl and 2.5 % v/v inoculum at pH 7.0 enhances the amylase production by 3.74-fold using Arthrobacteralpinus N16. Cold-active amylasecan be used in detergent, textile, food and beverage industries. Bio-degradation of starchy materials by cold active amylases can contribute in cleaning of environment at cold regions without harming the climate.

7 illus, 43 ref

024542   SHARMA S, KUMAR A, DOBHAL S, RAWAT A (Genetics Dep, Forest Research Institute, Dehradun-248 195, Email: ashok@icfre.org) : Standardization of protocol for genomic DNA isolation in Melia composita Benth. (Syn. M. dubia Cav.). Indian J Biotechnol 2018, 17(1), 185-8.

A simple, rapid and efficient method for extraction of genomic DNA from young leaves of Melia composita Benth. (Syn. M dubia Cav.) was optimized for obtaining fine quality DNA. The yield of extracted DNA ranged from 118.40 to 3192.40 µg /500 mg of tissue with absorbance ratio (A₂₆₀/A₂₈₀) ranging from 1.41 to 1.96. The quality of extracted DNA was also evaluated by agarose gel electrophoresis which showed a single intact band. The isolated DNA was free of any contaminating proteins, polysaccharides and coloured pigments and successfully amplify inter simple sequence repeat (ISSR) and simple sequence repeat (SSR) markers.

3 illus, 12 ref

024541   PAI S R, UPADHYA V, HEDGE H V, JOSHI R K, KHOLKUTE S D (ICMR National Institute of Traditional Medicine, Belagavi 590 010, 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 sterilized using both sodium hypochloride (NaOCl) (4.0 % v/v for 2 min) and mercuric chloride (HgCl₂) (0.1 % w/v for 2 min) solutions. In vitro plantlet regeneration in A. aspera was achieved from axillary explants cultured on Linsmaier & Skoog (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

024540   WANG J, LI X, ZHANG L, ZHANG Y, PENG Y, GUO M, ZHANG R, HUANG J (Minzu Univ of China, Beijing 100 081, 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

024539   CHAUHAN R S, JHA S K (Forest Biology and Tree Improvement Dep, Navsari Agricultural Univ, Navsari-396 450, 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

024538   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

024537   GAYACHARAN, BISHT I S, PANDEY A, YADAV M C, SINGH A K, PANDRAVADA S R, RANA J C (ICAR-National Bureau of Plant Genetic Resources, New Delhi-110 012, 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

024536   SABARA P, VAKHARIA D (Biotechnology Dep, Junagadh Agricultural Univ, Junagadh-362 001, Email: priteshrhsabara@gamil.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

024535   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

024534   ATRI C, SHARMA S, BANGA S S (Plant Breeding & Genetics Dep, Punjab Agricultural Univ, Ludhiana-141 001, 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

024533   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

024532   MYTHILI J B, CHETHANA B S, RAJEEV P R, GANESHAN G (Biotechnology and Plant Pathology Div, Indian Institute of Horticultural Research, Bangalore- 560 089, 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(1), 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

024531   JOSE A A, ANUSREE G, 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

024530   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

024529   VIGNESH V, SATHIYANARAYANAN G, PARTHIBAN K, KUMAR K S, THIRUMURUGAN 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

024528   ARORA V, GHOSH M K, SINGH P, GANGOPADHYAY G (Plant Biology Div, Bose Institute (Main Campus), Kolkata-700 009, Email: gaurab@jcbose.ac.in) : Light regulation of nitrate reductase gene expression and enzyme activity in the leaves of mulberry. Indian J Biochem Biophys 2018, 55(1), 62-6.

Nitrate reductase (NR) is considered to be an essential enzyme in plants, as it catalyses the conversion of nitrate to nitrite. In mulberry (Morus spp.), a plant of sericultural importance, the NR enzyme activity is found to be regulated by light. Three mulberry genotypes used in the present study showed that the NR enzyme activity increased in response to light. To identify whether light also regulates NR gene at the transcriptional level the expression of NR mRNA has been studied. The analysis of NR gene expression showed that light plays a significant role in regulating the NR gene with basal or no expression in response to light during the day while it is found to increase in the dark during the night showing maximum expression at pre-dawn hours.

3 illus, 21 ref

024527   JAISWAL A, SRIVASTAVA J P (Plant Physiology Dep, Banaras Hindu Univ, Varanasi-221 005, Email: ajaiswal308@gmail.com) : Changes in reactive oxygen scavenging systems and protein profiles in maize roots in response to nitric oxide under waterlogging stress. Indian J Biochem Biophys 2018, 55(1), 26-33.

Twenty days old pot grown seedlings of two genotypes of maize viz. HUZM-265 (waterlogging resistant) and HUZM-55 (waterlogging susceptible) were subjected to root zone waterlogging supplemented with 50, 500 and 2000 μmol/L sodium nitroprusside (SNP) as a donor of NO. Levels of reactive oxygen species (ROS) scavenging enzymes and non-enzymes were quantified in terminal root portions at 9, 12, 15, 18 and 21 h after waterlogging. In waterlogged plants, there was a significant increase in the levels of catalase (CAT), peroxidase (POX), superoxide dismutase (SOD), ascorbate peroxidase (APX) and alcohol dehydrogenase (ADH) and the increment was more in the resistant genotype. Levels of non-enzymatic antioxidants viz. ascorbic acid and phenols decreased, but to a lesser magnitude in the resistant genotype. After waterlogging, there was a differential pattern in variation in the levels of studied enzymes and non-enzymes with time. It is observed that ADH, CAT, and APX play a significant role to scavenge ROS at an early stage, while POX and SOD at later stages of waterlogging stress. In normal and waterlogged plants of HUZM-265, a low molecular weight protein of 20 kDa was visible, and at 500 and 2000 μmol/L SNP treatments another protein of 43 kDa appeared in this genotype under the influence of SNP. No such proteins were evident in HUZM-55. NO influenced studied parameters, but response varied with respect to genotypes, dose, and duration of waterlogging.

1 illus, 3 tables, 36 ref

024526   MAWLONG I, ALI K, TYAGI A (Biochemistry Div, Indian Agricultural Research Institute, New Delhi-110 012, Email: arunatyagi19@yahoo.com) : Functional validation of a water deficit stress responsive AP2/ERF family transcription factor-encoding gene in Oryza sativa. Indian J Biochem Biophys 2018, 55(1), 17-25.

Cloning of drought responsive genes and validating their function are essential to crop improvement. In the present study, a drought responsive AP2/ERF family transcription factor was isolated from drought-tolerant Oryza sativa L. cv N-22 (AP2/ERF-‘N-22’). Embryogenic calli produced in vitro from dehusked mature seeds of rice were bombarded with a gene construct containing AP2/ERF-N22, driven under inducible promoter RD29A from Arabidopsis using the Biolistic method. The bombarded calli were selected on hygromycin-containing selection medium. Molecular analysis of regenerated plants confirmed the integration and enhanced expression of the gene under water deficit stress (WDS). Transgenics showed 1.4 fold more expression as compared to wild-type (WT) under control condition and up to 2 fold more expression of AP2/ERF- ‘N-22’ under water deficit stress as compared to WT. Molecular analysis of regenerated plants confirmed the integration and enhanced expression of the gene under water deficit stress. The transformation efficiency was found to be nil, 0.97 % and 3.11 % for overstored seeds ≥ a year, ≥a year and fresh seeds respectively. Transgenics showed 1.4 fold more expression under control condition and up to 2 fold more expression under WDS as compared to wild-type (WT). About 90 % of the plants reached maturity and showed no negative phenotypic effects or aberrations as observed earlier under a constitutive promoter from that of the WT. Physiobiochemical analysis of transgenics showed enhanced drought tolerance.

4 illus, 1 table, 49 ref

024525   ROH I-S, KIM H-J, KIM H-J, SUH T-Y, HAN J-H, KANG H-E, SOHN H-J (Animal and Plant Quarantine Agency, Republic of Korea, Email: shonhj@korea.kr) : Polymorphisms in the prion protein gene, associated with chronic wasting disease, in the korean water deer (Hydropotes inermis argyropus). J Vet Sci Technol 2018, 8 (6), .

Prion diseases are fatal neurodegenerative diseases that affect humans and non-human mammals. Different alleles of the prion protein gene (PRNP) of humans and sheep are associated with varying susceptibilities to transmissible spongiform encephalopathies. Chronic wasting disease (CWD) is a prion disease of cervids, and polymorphisms at specific codons in the PRNP gene are associated with this disease. To assess the susceptibility of free-ranging deer to CWD, polymorphisms in PRNP were examined in the Korean water deer (Hydropotes inermis argyropus), focusing on codons that are potentially associated with CWD susceptibility (95, 96, 116, 132, 225, and 226). CWD surveillance was conducted by an antigen ELISA of tissue samples from 545 Korean water deer collected in eight provinces of the Republic of Korea. No prion protein associated with CWD was detected in any of the samples. These results suggest that PRNP of the Korean water deer shows low variation and the species has not been infected with CWD.

3 tables, 21 ref

024524   HAMIVAND Z, HADDADI G, FARDID R (Radiology Dep, Shiraz Univ of Medical Sciences, Shiraz, Iran, Email: rfardid@sums.ac.ir) : Expression of bax and Bcl₂ genes in peripheral blood lymphocytes of patients with differentiated thyroid cancer. J Med Phys 2018, 43(1), 41-5.

Thyroid cancer is the most common endocrine malignancy worldwide. Iodine‑131 is used in the treatment of thyroid cancer with dosage of 100 mCi. In the medical applications of ionizing radiation besides the advantages such as diagnosis and treatment of diseases, the risks arising from exposure should be considered as well. The present study aimed to evaluate the changes in expression levels of apoptotic Bax and Bcl‑2 and the ratio of Bax/Bcl‑2, in the peripheral blood lymphocytes (PBLs) of patients with differentiated thyroid cancer (DTC). This study was conducted on fifty thyroid cancer patients who had undergone surgery and were under treatment with 100 and 150 mCi doses. Blood samples were taken from the patients, one before iodine treatment and another 48 h after therapy. Bax and Bcl‑2 gene expression levels were measured by using real‑time reverse transcriptase polymerase chain reaction. The data were analyzed using one‑way analysis of variance followed by samples t‑test and independent samples t‑test. Significant changes were observed in the percentage of apoptotic cells, in groups, after radioiodine therapy compared with before treatment. The ratio of Bax/Bcl‑2 in both groups showed a significant increase (P < 0.001). The relative expression level of Bax gene showed a significant increase in comparison with the control group. Iodine therapy reduced expression of Bcl‑2 and a significant expression of Bax and finally increased the ratio of Bax/Bcl‑2. Iodine therapy led to apoptosis in the PBLs of patients with DTC. Therefore, it can be suggested that this method can be useful for monitoring and detecting destructive effects of ionizing radiation in nuclear medicine patients.

5 illus, 4 tables, 24 ref

024523   KULKARNI S S, PATIL S D, CHAVAN D G (Rajarshi Shahu Coll, Latur- 413 512, Email: deepa_dgc11@rediffmail.com) : Extraction, purification and characterization of hyaluronic acid from Rooster comb. J Appl Nat Sci 2018, 10(1), 313-5.

Hyaluronic acid, is extract by different procedures from various sources like pig, rabbit, oxes and human are available, but these processes have certain imitations like low yield, and also it requires the killing of these animals which is against the experimental ethics. In the present study, we have carried out the extraction of hyaluronic acid from cock’s comb which was further analyzed with qualitative test, viscosity, UV absorption, endotoxin detection assay. Also, the protein contamination of extracted hyaluronic acid was determined by using SDS-PAGE of hyaluronic acid was studied for checking the protein contaminants and it was noted that there were no bands observed in the well loaded with extracted hyaluronic acid sample indicating that the final extract of hyaluronic acid is not contaminated with the protein. The extraction and purification of hyaluronic acid by using the method reported here give pure hyaluronic acid. The viscosity of extracted hyaluronic acid was found to be 2.55 poise which is economical and can be used for industrial production of hyaluronic acid having clinical applications.

2 illus, 20 ref

024522   MAMO A, BALASUBRAMANIAN N (Adama Science and Technology Univ, Ethiopia, Email: nedumaran_b@yahoo.co.in) : Calf rennet production and its performance optimization. J Appl Nat Sci 2018, 10(1), 247-52.

Milk clotting enzymes are one of the most significant cheese making raw materials impacting and regulating milk coagulation activity. Calf rennet is one of the enzymes which extracted from suckling calf abomasum. The rennet extracted was optimized for its clotting time t (s) by varying three levels of temperature (30, 35 and 40 ^oC), pH (4, 5 and 6) and CaCl₂ concentration (0.1, 0.2 and 0.3 gm/500 ml of milk) using response surface method (RSM) and also its milk clotting activity (MCA) was determined. Based on the optimized result, temperature 39.13 ^oC, pH 4 and CaCl₂ concentration of 0.21 gm/500 ml had resulted minimum clotting time of 91.27 s. Using this minimum clotting time, the strength was found to be 1: 13148, establishing the fact that the calf rennet is the best natural coagulant of milk.

3 illus, 3 tables, 31 ref

024521   PARMAR J P, TIWARI R, GAUTAM K K, YADAV L, UPADHYAY N (Fruit Science Dep, Horticulture Coll, Mandsaur- 458 001, Email: rt_jnkvv@yahoo.com) : Effect of Indole 3-butyric acid (IBA), rooting media and their interaction on different rooting and growth characteristic of air-layers in guava (Psidium guajava L. cv. L-49). J Appl Nat Sci 2018, 10(1), 241-6.

The present investigation was undertaken to study the effect of different concentrations of Indole 3-butyric acid (0, 2000, 4000, 8000 and 10000 ppm), different types of rooting media (i.e. sphagnum moss, coco peat, vermicompost, sphagnum moss + coco peat and sphagnum moss + coco peat + vermicompost) and the interaction of these in different combinations on rooting and growth characteristic of guava, cv. L-49. The results revealed that the exogenous application of Indole 3-butyric acid (IBA) 8000 ppm with media combination of sphagnum moss + coco peat + vermicompost (I₃M₅) significantly increased the root characters i.e. success percent of air-layers (83.3 %), callus formation (23.7), number of primary roots (24.21) and secondary roots (32.57); and growth characters i.e. number of leaves (48.17), branches (7.92) and sprouts (8.48) and survival percentages (83.18) over the other treatments and control. Among the rooting media, the combination of sphagnum moss + coco peat + vermicompost (M₅) produced the highest rooting percentage (71.27), root characters and growth characters during months of July and August.

2 tables, 27 ref

024520   MALINI N, ANANDAKUMAR C R, GNANAM R, RAMAKRISHNAN S H (Plant Breeding and Genetics Dep, Tamil Nadu Agricultural Univ, Kovilpatti - 624 501, Email: malinipbg200201@gmail.com) : Effect of harmones on callus induction in maize (Zea mays L.). J Appl Nat Sci 2018, 10(1), 202-9.

Callus induction from explants is a critical process in regeneration, micropropagation and transformation of maize (Zea mays L.) plants. Formation of callus from plant tissues on culture is affected by several factors. This study revealed to establish the effect of genotype, source of explants and auxin concentration on callus induction from five genotypes UMI 757 (G₁), UMI 615 (G₂), UMI 112 (G₃), UMI 285 (G₄) and CO 1 (G₅) and one hybrid CO H (M) 5 (G₆). Callus induction of the six maize varieties was investigated using immature embryos (E₁), leaf bits (E₂), root tips (E₃), hypocotyls (E₄) and seeds (E₅) as explants with different concentrations of hormones. In this study, immature embryo was taken from 10 to 12 days after pollination (DAP) to get maximum response. The highest percentage of callus induction was observed (99.10) in immature embryo culture and seed culture gave the highest percentage of rhizogenic callus formation when compare to immature embryo. Among the genotypes tested, CO H (M) 5 recorded the highest callus induction percentage on (2D2K2) medium composition.

3 tables, 25 ref

024519   KURIKESU I, ANUJA T J, GANGAPRASAD A, NAIR A J (Biotechnology Dep, Kerala Univ, Kerala-695 581, Email: jayakumarannair23@gmail.com) : Regulation of microRNAs during biotic and abiotic stress. Bull Pure Appl Sci- Sect B 2018, 37B(1), 49-55.

MicroRNAs (miRNAs) are short non-coding RNAs approximately 22 nucleotides play key roles in fundamental cellular processes, comprising how cells react to changes in environment or, stress. Stress is the state when cells deviate from the current situation due to sudden environmental changes or frequent fluctuations in environmental factors. Apart from the cellular concentration of miRNAs, the target gene repression level is also dependent on the concentration of mRNA target relative to the miRNA. From the previous studies it is seen that many aberrant expression of many miRNAs is induced by abiotic stressors so that miRNAs can be used as a new target for genetically improving plant tolerance to many stress. miRNAs respond to environmental stress mainly tissue and genotype-dependent manner. During abiotic stress, miRNAs function by regulating target genes within the miRNA-target gene network and by controlling signalling pathways and root development. In this review, we revisit the processing of microRNAs in animals and plants, history of miRNAs and condense recent findings in miRNA biogenesis and microRNAs involved in biotic and abiotic stress.

51 ref

024518   LAKHERA K, KUMAR A, RANI A, DIXIT R, RANA S (Biotechnology Dep, Swami Vivekananda Subharti Univ, Uttar Pradesh- 250 005, Email: lakhera.kanchan@gmail.com) : Plant tissue culture and its application. Bull Pure Appl Sci- Sect B 2018, 37B(2), 32-6.

Plant tissue culture has been successfully integrated into plant transformation strategies. Meristem culture has become immensely popular as micro propagation technique for the rapid production of commercially viable plants at industrial level. Meristem culture is also significant for obtaining virus free plant and additionally, meristem culture in combination with other techniques is applied in plant improvement as in production of transgenic plants, plant germplasm storage, regeneration of quality planting material or seeds in association with aeroponic or hydroponic systems.

17 ref

000622   DIAB E, BAZID A-H I, FAWZY M, EL-ASHMAWY W R , FAYED A A, EL-SAYED M M (Virology Dep, Suez Canal Univ, Ismailia 41522, Egypt, Email: mohamed_mohamed4@vet.suez.edu.eg) : Foot-and-mouth disease outbreaks in Egypt during 2013-2014: Molecular characterization of serotypes A, O, and SAT2. Vet World 2019, 12(2), 190-7.

Foot-and-mouth disease virus (FMDV) serotypes A, O and South African Territories (SAT2) are endemic in Egypt; each is presented by a number of partially related topotypes and lineages, depending on their geographical origin. Continuous mutations and the emergence of new topotypes that lead to occasional vaccination failures were frequently recorded, so this study aimed to genetically characterize the circulating FMD virus strains in Egypt during 2013 and 2014 outbreaks, focusing on amino acids variations in VP1 region. A total of 51 oral tissue samples were collected from cattle and buffaloes in 13 farms, and 38 individual cases showed clinical signs suspected to be FMD in six Egyptian Governorates (Cairo, Giza, Qaliubia, Fayoum, Sharquia, and Assiut). FMDV in collected samples was characterized by reverse transcription-polymerase chain reaction (RT-PCR) amplification of full VP1 region, sequencing, and phylogenetic analysis. Out of 51 samples, 44 (86.27 %) were positive by RT-PCR using universal primers. Serotype O was predominant and detected in 31 samples (70.45 %), serotypeA was detected in 9 samples (20.45 %), and then serotype SAT2 was identified in 4 samples (9.10 %). Sequencing and phylogenetic analysis of VP1 demonstrated clustering of serotype O, A, and SAT2 in EA-3 topotype, ASIA topotype, and topotype VII, respectively. Serotype O is closely related to O/SUD/8/2008 with 94.6 % identity but showed 14.6 % differences from vaccine strain (O/PanAsia-2) of ME-SA topotype. Furthermore, Serotype A and SAT2 were closely related to recent circulating Egyptian isolates and vaccine strains type A/EGY/1/2012 (Asia topotype, lineage Iran-05) with identity 96.4 % and vaccine strain of SAT2/EGY/A/2012 (topotype VII, lineage SAT2/VII/ALX-12) with identity 95.3 %, respectively. The present study recommended further studies of serotype O to determine the immunogenic relationship between the vaccine strain and the new strains to attain maximum protection against circulating viruses.

5 illus, 2 tables, 32 ref

023564   DEHINWAL A K, PAHUJA S K, JOSHI U N, KUMARI P, ARYA S (Genetics and Plant Breeding Dep, CCSHAU, Haryana- 125 004, Email: ashokdehinwalccshau@gmail.com) : Study of combining ability for quality component in Forage Sorghum [Sorghum Bicolor (L.) Moench]. Biosci Biotechnol Res Asia 2017, 14(4), 1533-42.

The study envisaged assessing the general combining ability of the parents and specific combining ability of the hybrids, using line x tester mating design. Twenty four hybrids along with their parents and checks ((SSG 59-3 and MFSH 4)) were evaluated at two locations with two date of sowing (Early and late sowing) during the kharif season of 2015-16. Data on five randomly taken plants from each genotype in each replication were recorded on different quantitative characters at first cut (55 days after sowing) and second cut (45 days after first cut). The ratio of ó2 GCA/ó2 SCA was less than unity for all the characters indicating preponderance of non-additive gene action (dominance and epistasis). Female parents 9A and 56A were also better combiners for HCN content, IVDMD and DDM in more than two different environments. HJ 513 and G 46 were found to be good general combiner male parents for protein content, protein yield, IVDMD and DDM in more than two different environments. The Cross combination of 465A × HJ 513 and 9A × IS 2389 were better for protein yield, IVDMD and DDM in more than two different environments. This suggests the usefulness of heterosis breeding or any breeding plan which makes use of specific combining ability effects for improvement in these traits.

6 tables, 13 ref

023563   TEHRANI M M, SOBHANI M (Technical and Engineering Dep, Islamic Azad Univ, Shahroud, Iran, Email: mahdipourtehrani96@gmail.com) : Synthesis of bioactive glass powder using Sol-Gel method and shaping using plasma spark sintering. Biosci Biotechnol Res Asia 2017, 14(4), 1523-31.

In the present study, the synthesis of bioactive glass powder is carried out using the sol-gel method and in an acidic environment. TEOS (Tetraethyl orthosilicate) is used as the raw material of supplying Silica and TEP (three ethyl phosphates) is used as the raw material for providing phosphate powder compounds. Synthesis is done without water and using pure alcohol solvent 99.99%. After preparation of powder from the gel and drying it, the heat treatment process was performed at 500, 550 and 600 C for 2 hours on the powder. Thus, the bioactive glass powder was prepared for the SPS stage. In the present study due to technological limitations (impossibility of access to the high pressures of the SPS device used), at the formation stage, the maximum force of 20 tons was selected using the SPS device. Experiments performed on the powder include XRD and DTA. The results of this research show that performing heat treatment at a maximum temperature of 550 ° C with the aim of increasing the maintenance of the amorphous structure of the product is the most ideal temperature for heat treatment. Also, the use of suitable molds in the SPS process with a tolerance of up to 300 MPa is a requirement of the process.

4 illus, 7 ref

023562   HAIDER M J, AHMED D S, MOHAMMAD M R, HAIDER A J (Applied Science Dep, Univ of Technology, Baghdad, Iraq, Email: duhasaadi2015@gmail.com) : Modification of functionalized multi walled carbon nanotubes by olive oil as economic method for bacterial capture and prevention. Biosci Biotechnol Res Asia 2017, 14(4), 1513-22.

In the present work, a new and economic method of functionalized Multi walled Carbon Nanotubes (MWCNTs) with olive oil has been improved as compared to the traditional methods, that composed of catalysts such as fatty acids (FFA), glycerol and triacylglycerols of oxidation power. This method is initiated by ultrasoniction the mixture which leads to functionalize the surface during the process of interaction with oil olive for a short time (30 min). Then, the results of this work were examined by using Scanning Electron Microscopy (SEM) and High Resolution Transmission Electron Microscopy (HRTEM) to study the morphology of the surface treated with olive oil, X-ray diffraction (XRD) was employed to confirm the functionalized method and using Fourier transformed infrared spectroscopy (FTIR) that shows the formation of functional group on to MWCNTs surface such as C=O and COOH. The final part includes a study antibacterial activity of Multi walled Carbon nanotubes (MWCNTs) against gram negative (E. coli and Salmonella typhi) and gram positive (S. aureus) bacteria using direct cell counting. Besides, evaluating the antibacterial assays of treated MWCNTs after incubated for various time to elucidate the interaction between functionalized MWCNTs and cell membrane to determine the study the adsorption of bacteria on the surface of (MWCNTs), which are used in environmental applications, such as sensing of contaminated water and using as a filter.

11 illus, 27 ref

023561   DAYAL S, KUMAR H (Biotechnology Dep, Monad Univ, Hapur, Email: harshal78617@gmail.com) : Assessment of genotoxic effects of Nux Vomica ashomeopathic drug on mitotic chromosomes of Vicia Faba. Biosci Biotechnol Res Asia 2017, 14(4), 1497-1502.

Extract of Nux vomica is used as homeopathic drug to cure various nervous disorders like mental emotion epilepsy, prolepses of the rectum, hydrophobia etc. The given dose of this drug is very small because of its poisonous effect in higher dose. In the present investigation we want to assess toxic effects of Nux vomica on somatic chromosomes of Vicia faba. Four concentrations 5%, 10%, 20% and 30%, were used for the treatment of root tips of test plant. There were number of abnormalities observed like stickiness at orientation of chromosomes, unorganization of chromosomes, fragmentation during separation of chromatids, multiple chromatin bridges formation etc. These types of aberrations were increased with increased concentration and duration of treatment of this drug.

1 table, 24 ref

023560   KURIEN N K, BHAT S G (Biotechnology Dep, Cochin Univ of Science and Technology, Cochin- 22, Email: saritagbhat@gmail.com) : Photoprotection and anti-inflammatory properties of non–cytotoxic melanin from marine isolate Providencia rettgeri strain BTKKS1 Providencia rettgeri strain BTKKS1. Biosci Biotechnol Res Asia 2017, 14(4), 1475-84.

Photoprotection and Anti-inflammatory properties of characterized melanin produced by marine proteobacterium Providencia rettgeri strain BTKKS1 was explored in the study. Characterization of melanin was carried out by chemical, FTIR, proton NMR and EPR analysis. The radical scavenging property was estimated using DPPH assay and Fe²⁺ chelating potential was also evaluated. Effect of melanin on the activities of Cyclooxygenase, Lipoxygenase, Myeloperoxidase and Cellular Nitrite is used to evaluate anti-inflammatory potential. Enhancement of Sun Protection Factor (SPF) is evaluated to study its effectiveness in photoprotection. Cytotoxicity of melanin was estimated using MTT assay.The chemical, FTIR, proton NMR and EPR characterization were typical of eumelanin. The pigment also showed profound radical scavenging activity (63.73%) and metal chelating potential (97.09%). Melanin significantly inhibited the activity of the inflammatory enzymes in a dose dependent manner and enhanced the SPF value of commercial sunscreens at an average of 2.64 factors. This melanin was also less cytotoxic with an IC50 value of 97.87µg/mL. The immense Anti-inflammatory property of the pigment can be utilized in therapeutic applications. The photoprotection potential of melanin can be utilized in cosmetic formulations, UV protection devices etc.

6 illus, 1 table, 53 ref

023559   TASLEEM M, BAUNTHIYAL M, GOHARTAJ (Biotechnology Dep, G. B. Pant Institute of Engineering & Technology, Garhwal, India., Email: mohdtasleem99@gmail.com) : Induction of MPK3, MPK6 and MPK4 mediated defense signaling in response to Alternaria Blight in transgenic Brassica juncea. Biosci Biotechnol Res Asia 2017, 14(4), 1469-74.

Alternaria brassicae causes a highly destructive disease in Brassica juncea (Rapeseed mustard) resulting in significant yield losses. Studies of MAPK machinery components in Arabidopsis thaliana have indicated that MPK3, MPK4, & MPK6 are involved in defense response and provide resistance against various bacterial and fungal pathogens. In this study, we analyzed the expression level of MPK3, MPK4 & MPK6 in overexpressed MPK3 transgenic (BjV5) Brassica juncea at different stages of Alternaria brassicae inoculation.Expression study revealed that MPK3/MPK6 was involved in early defense response and MPK4 in late defense response. These results suggested that BjMPK3 positively regulate SA mediated defense response, which might play an important role in resistance to Alternaria brassicae in Brassica juncea.

2 illus, 1 table, 25 ref

023558   NAIR D N, PADMAVATHY S (Bioinformatics Dep, Nirmala Coll for Women, Tamil Nadu- 641 018, Email: dhanya.dnn@outlook.com) : Molecular docking studies of phytocompounds from Aloe vera (L.) Burm.f. having anticancer property, against an antiapoptotic Bcl-2 protein. Biosci Biotechnol Res Asia 2017, 14(4), 1449-56.

B-cell lymphocyte-2 (Bcl-2) is an antiapoptotic protein, which is an important member of Bcl-2 family. The current study involves molecular docking of six antineoplastic phytocompounds from Aloe vera (L.) Burm.f. against the protein Bcl-2. Docetaxel, a known inhibitor of Bcl-2 was used as a control in this study. All the studied phytocompounds bound within the same binding pocket as that of Docetaxel and thus can be considered as potential inhibitors of Bcl-2 protein. Among the six phytocompounds studied, AVG4 showed the best docking result, with a minimum pharmacological energy, -198.9 kcal/mol, followed by AVG6 and AVG3 as the second and third best phytocompound while AVL3 has the maximum pharmacological energy -103.8 kcal/mol. AVL3 is involved in cation-pi interactions with the Tyr9 residue of the Bcl-2 protein which is not considered while calculating pharmacological energy scoring function. Calculation of energy due to cation-pi interactions may result in the increase in total binding energy of AVL3, which may significantly increase the pharmacological energy, E_Pharma by approximately -8 kcal/mol, resulting in another potential anticancer phytocompound.

5 illus, 3 tables, 25 ref

023557   LEELA K, DEVI C A (PG & Research Dep of Biotechnology, Women’s Christian Coll, Chennai- 600 006, Email: dr.anchanababu@gmail.com) : Isolation, purification and application of secondary metabolites from Lichen Parmelia Perlata. Biosci Biotechnol Res Asia 2017, 14(4), 1413-28.

Lichens are composite algae having a symbiotic association with a fungal partner. They produce numerous secondary metabolites, which play an important role in pharmaceutical and in other industrial applications. The Secondary metabolites produced by lichens are found to be 80% more when compared to that produced by other organisms. Not much work has been carried out on lichens due to the difficulty in their cultivation but still it emerges as a potential source in developing therapeutically important drugs which are widely beneficial in all fields of application. The present study was aimed to isolate, purify and determine the applications of secondary metabolites from Lichen Parmelia perlata. The presence of these compounds were detected and purified by thin layer chromatography and column chromatography using specific solvent systems. The purified fractions were then identified by Gas chromatography-Mass spectrometry (GC-MS). The compounds were then subjected to application oriented studies such as antimicrobial activity, antioxidant activity and antidiabetic activity. Not much work have been carried out on the isolation of a specific glycoside and alkaloid compound from Lichen Parmelia perlata, so this study was an attempt to explore the applications of these individual compounds which could prove beneficial to the mankind for different purposes.

12 illus, 4 tables, 35 ref

023556   TARIVARDI Z, NUGHABI K A, PIRI S (Biotechnoly and Molecular Genetics, Islamic Azad Univ, Zanjan, Iran, Email: zahratariverdi96@gmail.com) : Propagation of rootstocks of Gisela 5 based on tissue culture method. Biosci Biotechnol Res Asia 2017, 14(4), 1395-9.

Cherry is one of the most common fruits in the moderate conditions. Common rootstocks like Gisela5 have been used in the gardening field in recent years. The purpose of the current study is investigating the enhancement of Gisela5 rootstock method on the basis of tissue culture. Cutting grafting was used as explant material. These explants included the following materials in the MS environment: 1mg/l IBA (Indole-3-butyric acid), 0.75 mg/l BPA, 1mg/l BPA+ 1 mg/l BPA, 0.75 mg/l BPA + 2mg/l BPA and 1mg/l BPA +2mg/l BPA. The planting was kept in the planting room with 24 ± 2 ^°C temperature. The basic environment of MS completed with 0, 1,2,4,6 mg/l NAA during the rooting stage. The maximum number of branches which was 289 was achieved by means of the combination of the following values: 1 mg/l INA+ 0.75 mg/l BAP and 2mg/l IBA +1 mg/l BAP. The maximum length of the branches was 1.69 cm. Varieties of concentrations of NAA were added in the rooting stage. The amount of rooting was increased by adding almost 6 mg/l of NAA into the rooting stage.

2 tables, 19 ref

023555   SINGH Y, TIWARI A, KUMAR R, SAXENA M K (Veterinary Physiology & Biochemistry Dep, G.B. Pant Univ of Agriculture & Technology, Pantnagar- 263 145, Email: mumtesh@rediffmail.com) : Cloning, sequencing and phylogenetic analysis of stn gene of Salmonella Typhimurium. Biosci Biotechnol Res Asia 2017, 14(4), 1387-93.

Salmonella Typhimurium is an important facultative bacillus pathogen having broad host range and high physiological adoptability. Several genes contribute in virulence and determine pathogenicity of this isolate. Stn is an important virulent gene which code for Salmonella toxin, increases the level of c-AMP in the host, and ultimately results into diarrhoea and vomiting. In present study stn gene was cloned, sequenced and on basis of sequence information of stn gene, phylogenetic relation was deduced between different serovars of Salmonella Typhimurium. Genomic DNA was isolated from field isolate of Salmonella Typhimurium (isolate No-A201) and stn gene was amplified using gene specific primer and cloned in pJET vector by the positive selection system. Amplification of stn gene yielded a product of approximately 750 bp. Subsequently gene was sequenced and a complete ORF of 750 bp was obtained. The sequence was submitted to NCBI Genbank and allotted the Accession No KF032246 was allocated. Sequence was further used for bioinformatics analysis of Stn protein, which exhibited two major domains and one amino acid substitution at 609 residue. On phylogenetic analysis, S.Typhimurium exhibited 99% similarity with Salmonella enterica subsp. enterica serovar Newport. Our findings indicate that stn is an important toxin gene, which is conserved among many serovars of Salmonella.

6 illus, 26 ref

023554   RASHIDI S, SANI A H, RAZAVI S (Chemical Engineering Biotechnology, Islamic Azad Univ, Tehran, Iran) : The effect of environmental conditions on the removal of organic compounds from color sludge via bioslurry method. Biosci Biotechnol Res Asia 2017, 14(4), 1371-85.

Every industrial activities resultes in the pollution of environment. This pollution could be in the shape of voice, weather, and water or earth pollution. The amount of pollution in industries is different and it depends on the type of industry. So to purify and remove pollution, should know accuracy the amount and the type of pollutants. The sewage in the automotive industry includes 2 parts: Paint sludge produced in the automotive industry has the highest pollutant capability because of weighty elements. So, the removal of it in the wrong way will cause damage to the environment. In this research, a number of experiments were carried out to identify the main pollutants in the paint sludge. In this research, the method of research and data gathering was the use of a slurry bioreactor with the capability of the regulation of the amount of aeration and mixing speed to remove organic compounds of paint sludge. The experiment was designed according to the Taguchi method, which 9 experiments were designed to achieve the desired result. The results of the two experiments showed a significant reduction in the initial COD of 76 and 80% biologically pure and it became clear that aerobic bacteria purify the paint sludge more. Also, after analyzing all the results of Mini-Soft software, optimal purification conditions for effective environmental parameters were pH = 7, stirring speed = 14 rpm, dissolved oxygen concentration = 5.5 ppm. Finally, after the activation of the reactors to filter 1 kg of paint sludge for 30 days and at ambient temperature, the result of the two experiments showed a significant reduction in the initial COD of the biological treatment of 76 and 80%, and it was found that the aerobic bacteria do more the color sludge purification. Also, after analyzing all the results with mini-tab software, the optimum refining condition for effective environmental parameters was pH = 7, active sludge = 0.5 kg, stirring speed = 14 rpm, dissolved oxygen concentration = 5.5 ppm.

13 illus, 5 tables, 15 ref

023553   SUNDARAM B, MILTON M C J (P.G. and Research Dep of Advanced Zoology and Biotechnology, Loyola Coll, Tamil Nadu, Email: biotechres17@gmail.com) : Porous polycaprolactone scaffold engineered with naring in loaded Bovine Serum Albumin nanoparticles for bone tissue engineering. Biosci Biotechnol Res Asia 2017, 14(4), 1355-62.

A biodegradable three-dimensional scaffolds have gathered attention and are widely studied for bone tissue engineering applications. In the present study, porous polycaprolactone scaffold entrapped with naringin loaded bovine serum albumin nanoparticles (PS-N-BSANP) has been engineered. Further, the prepared nanoparticles and interconnected porous scaffolds were characterized by scanning electron microscopy, X-ray diffraction and fourier transform infrared spectroscopy analysis. X- ray diffraction showed amorphization of naringin in PS-N-BSANP. In addition, sustained naringin release profile was observed from PS-N-BSANP for 12 days which showed a cumulative release of 52.54 micromolar (µM). Furthermore, conditioned medium from PS-N-BSANP showed an increased calcium deposition and collagen matrix formation under osteogenic conditions with C3H10T1/2 cell line. These results suggest that PS-N-BSANP enhanced the osteogenic differentiation potential in bone tissue engineering applications due to the controlled release of naringin.

6 illus, 33 ref

023552   GURUMURTHY B R, NATARAJU S P, PAVITHRA S K (Crop Physiology Dep, Univ of Agricultural and Horticultural Sciences, Shivamogg, Email: shimogagurumurthy@gmail.com) : Variability in polyphenol content in Areca nut(Areca catechu) samples of Karnataka. Biosci Biotechnol Res Asia 2017, 14(4), 1343-7.

The phenolic compounds or polyphenols, secondary metabolites of plants, are phytochemicals that exhibit antioxidant activity and consequently possess a beneficial physiological effect (Bravo, 1998). Plant polyphenolic compounds have been recognised for their ability to prevent oxidation of susceptible substances by virtue of their electron donating property due to presence of large number of phenolic hydroxyl groups. The seed of Areca catechu contains higher proportions of polyphenolic compounds mainly tannins (Zhang et al., 2008). In India 7.06 lakh ton of arecanut is produced annually (2016-17), major portion of which (App, 90%) utilised for human consumption. Hence it is essential to study variability in polyphenol contents of areca samples grown under diverse conditions and its impact on consumers. In the present study 850 areca samples were collected from 6 important area growing districts of Karnataka and further hobliwise pooling was done before polyphenol content was determined. In Shimogga district high concentration of polyphenol was found in Bhadrvathi thaluk (3.02%) followed by Shimogga (2.68%) and less concentration was found in Sorab taluk (1.17%). Polyphenol content varies from 0.82 % to 2.14 % in Chikkamagalur. The highest concentration was observed in Tarikere (2.14%) and less in Kadur (1.55%) . In Davanagere district polyphenol content was highest in Channagiri (2.50%) followed by Jagalur (1.29%). The results of total polyphenol content in different district are very different. But the region wise variability was not noticed.

3 tables, 6 ref

023551   SANDINO J D, HURTADO D A, RAMOS O L (Virtual Applications Group, Univ Militar Nueva Granada, ) : Prediction of reproductive system affectation in Sprague Dawley rats by food intake exposed with Fenthion, using Naïve Bayes classifier and genetic algorithms. Biosci Biotechnol Res Asia 2017, 14(4), 1291-7.

Improper application of pesticides in agricultural crops and indirect effects caused by exposure to them through consumption of contaminated crops, nowadays represent a serious risk to public health harmony. It is vital then, to know the degree of toxicity of each of these chemicals in order to properly regulate its application and sensitize the population at risk. Therefore, this paper shows the results of an algorithm with the ability to predict the effects on the reproductive system in Sprague Dawley rats, caused by the intake of food exposed with Fenthion. The original data were processed using the Naïve Bayes classifier, then optimized using genetic algorithms. It is concluded that the prediction algorithm does the job properly, processing qualitative information with relatively low computational cost, which allows its easy portability to different development platforms.

3 illus, 2 tables, 25 ref

023550   RITAMBHARA, KUMAR A, SRIVASTAVA D S L, ,VIJAYARAGHAVALU S, KUMAR M (Biochemistry Dep, Univ of Allahabad, Allahabad- 211 002, Email: munishkp@gmail.com) : GSTM1/GSTT1 gene polymorphism in North Indian population and their association to hypertension. Biosci Biotechnol Res Asia 2017, 14(4), 1269-75.

Hypertension has multifactorial background and is a life style related disorder. Arterial hypertension is one of the most common types of hypertension and associated to oxidative stress known to be implicated in its pathogenesis. Glutathione S-transferases (GSTs) are phase II detoxification enzymes that play an important role in cellular defense against oxidative. GSTT1 and GSTM1 gene polymorphisms are reported to be associated with arterial hypertension in several populations and ethnic groups. GSTM1and GSTT1null genotype results in the impaired enzyme activity and consequently greater vulnerability towards various diseases such as cancer. The present study has been done to assess whether GSTT1 and GSTM1 null genotypes are associated with arterial hypertension among North Indian population. We have enrolled 99 arterial hypertension patients and 99 age and sex matched control individuals. Genotyping of GSTM1 and GSTT1 was done by multiplex PCR. We found that GSTM1 null genotype may have risk for hypertension (OR 1.75, CI 0.93-3.2), while for GSTT1 null genotypes shows protection toward hypertension (OR 0.254, CI-0.113-0.57). GSTM1 null genotypes serve as potential genetic factor and may be an independent risk factor for development of hypertension whereas GSTT1 null may have role in protection against hypertension in North Indian populations.

1 illus, 4 tables, 39 ref

023549   SULAIMAN O O (School of Ocean Engineering, Univ of Malaysia, Malaysia., Email: o.sulaiman@umt.edu.my) : Model test for determination of hydrodynamic ocean coeficient for design of aquaculture mooring system for oceanic macroalgae farming. Biosci Biotechnol Res Asia 2017, 14(4), 1227-34.

Ocean and Coastal farming has potential economic benefits as a source of sustainable. Prototype plantations have been deployed in coastal areas and therefore are competing for limited space with other shoreside activities such as recreation and industry. If seaweed can be cultivated in relatively deeper waters away from populated areas, production may be expanded using larger areas not presently exploited for other purposes. Prototype systems aquaculture plantations have so far relied on ad hoc, field developed mooring systems, rather than engineering design principles. Such systems may not be sufficiently reliable for deepwater deployments, where repairs are more costly and resources are scarce. A quickest way to obtain design loadings for seaweed would be to carry out model tests on some available seaweed samples.The main difficulty is that the flow kinematics (fluid velocities/accelerations) that can be produced in the available model basin, at the Marine lab of the Universiti Teknologi Malaysia in Johor Bahru, where the tests are to be carried out, are suitable only at smaller scale. This paper describes the model tests and analysis used to develop the loads needed so that the mooring system for the aquaculture system can be designed based on methods like those used for offshore structures.

10 illus, 3 tables, 8 ref

023548   SULAIMAN O O, GUERRIERO G, AKHIR F, NABILAH I (School of Ocean Engineering, Univ Malaysia Terengganu, Malaysia, Email: oolanrewaju8@gmail.com) : Nutrient and oceanography requirement for G. Edulis and U. Lactuca in Setiu wetland. Biosci Biotechnol Res Asia 2017, 14(4), 1221-6.

Nutrients and Oceanography parameters are the key environmental factors that can determine biomass yield and productivity of an aquaculture system. Oceanography parameters are primarily controlled by the choice of location and season of the aquaculture site. Nutrients are controlled on the site and nutrient manipulations can control biomass yield, productivity and the amount of certain products formed. Carbon, nitrogen and phosphorus are the main three nutrient elements that influence the growth of seaweed. The different concentration of nutrients at difference aquaculture site was determined which in return influence types of seaweed growth at certain location.

9 illus, 3 tables, 4 ref

023547   SHARMA S, SINGH G, SHAIK S P, SINGH G, BHAT S, SHARMA G (Natural Sciences Dep, Sant Baba Bhag Singh Univ, Jalandhar- 144 030, Email: g.sharma7346@gmail.com) : Gene stacking for fungal resistance in plant transformation vector. Biosci Biotechnol Res Asia 2017, 14(3), 1211-9.

Fungal diseases like early blight, late blight, fusarium wilt cause 30-40 per cent loss in fruit production. Form past decade many transgenic plants had been developed using genes encoding chitinases and glucanases with the objective of imparting fungal disease resistance. Since the genes encoding chitinase and glucanase act synergistically. The study was performed to construct plant transformation vector pRAGS carrying both ech42 and bgn under single T-DNA region. Initially, HindIII site at 5' end of earlier cloned bgn (T. harzianum) was removed using primers during reamplification of the gene. The amplicons were cloned into pTZ57R/T containing T overhangs at Eco321 site and transferred to E. coli DH5a and further to plant transformation vector pBI121 which was named as pRA121. In order to clone another gene (ech42) into pRA121, expression cassette from iHP vector was transferred to pRA121 and named as pRAG121. Further in order to gain XhoI site for cloning ech42 gene into pRAG121, ech42 (pSUM1) was cloned into pYES2/CT, named as pSAG1, ech42 from pSAG1 cloned with KpnI and XhoI in pRAG121 and named as pRAGS121. The vector constructed in the present study can be used to transform important crop plants to have enhanced resistance to fungal diseases.

10 illus, 36 ref