025539   AL-GHUFAILI M K F, AL-TAMIMI A J T (Kufa Univ, Iraq, Email: atyaf.altameemi@uokufa.edu.iq) : Genetic relationship among ten wheat genotypes using seventeen RAPD markers. Plant Arch 2018, 18(1), 595-600.

This study was conducted for determination DNA fingerprint and estimation of genetic diversity among ten wheat (Triticum aestivum L.) genotypes using seventeen RAPD Markers. Primers OPB-06, OPC-05, OPH-01 and UBC-126 gave unique fingerprint for studied genotypes. Primer OPG-09 gave higher value for polymorphism . The higher efficiency and discriminatory value was produced by primer UBC-126. High genetic distance was 0.546 between Buhuth22 and Faris while low genetic distance was 0.142 between Rasheed and Iraq. Cluster analysis (Phylogenetic tree) by UPGMA based dendrogram revealed that studied genotypes grouped in two main clusters. Low polmorphisim (36.827) revealed by studied primers. Results show that RAPD markers could efficiently reveal genetic variation and fingerprint wheat genotypes .

5 illus, 4 tables, 38 ref

025538   JWAD R A, RUBAIEE H M A, KHALIL F A A, KHALEEL A I (Plant Protection Dep, Kufa Univ, Iraq, Email: rashamohammed2011@gmail.com) : Genetic diversity assessment of some stored insect species in Iraq based on RAPD molecular marker. Plant Arch 2018, 18(1), 546-50.

Genotyping of four insect species was carried out using RAPD marker. The genetic variability among the four insect species was estimated using RAPD primers. All primers generated reproducible and easily storable RAPD profiles with some amplified DNA fragments ranging from 9 to 13. The total number of amplicons detected was 56, including unique bands, reached seven, Number of polymorphism was 49 and this represents a level of polymorphism of 87.04 % and an average number of 6.8 polymorphic bands per primer. Maximum numbers of amplicons were amplified by primer U-17 reached 13 while the minimum number of fragments was amplified with primers OPE K-02 reached nine. The highest number of polymorphic bands reached 12 were obtained with primer U-17, while the highest number of monomorphic bands reached two was obtained with primer OPE K-02, and M-32 with percentage reached 22.22 % and 16.664, respectively. RAPD markers detected genetic similarity and distance, a maximum genetic distance value was observed between T. granarium(1) and C. maculatus(4) and S. oryzae(2) and C. maculatus(4) species reached 0.547 with less similarity value reached 46 %, a minimum genetic distance value was 0.435, which observed between S. oryzae (2) and T. castaneum (3) with high similarity value reached 57 %. The similarity matrices were employed in the cluster analysis to generate a dendrogram using the UPGMA method. The cluster tree analysis showed that the insect species were broadly divided into two main groups A and B with genetic similarity reached 30 %. A group including T. granarium (1) B group was divided into two sub-cluster B1, and B2 with genetic similarity reached 36 %. The first sub-cluster (B1) including only C. maculatus (4) while the second sub-cluster (B2) included two species S. oryzae (2) and T. castaneum (3) with the high genetic similarity between them reached 48 %.

3 illus, 2 tables, 17 ref

025537   ABFULHUSSEIN F R, MUTLG N H, SARHEED A F (Field Crops Dep, Al-Muthanna Univ, Iraq) : Genotypic characterization and tissue localization of the mutant lines expression of OsHKT1;3 gene in rice under salt stress. Plant Arch 2018, 18(1), 489-95.

In rice (Oryza sativa ‘Nipponbare’), the genes family HKT is composed of eight members. It encodes for Na⁺ transporters that play an important role in salt stress tolerance. Functional analysis, it has been determined that there is a wide diversity of these transporters in terms of their Na⁺/K⁺ selectivity and K⁺ affinity, in addition, plays an important role in reducing Na⁺ accumulation in shoots to cope with salt stress. This study focuses on one of these genes, yet to be characterized in literature: OsHKT1;3 (also known as HKT6). Using reverse genetics, the aim of this study is to determine the role of this gene during salt stress. In order to observe the expression of this gene in different organs of the plant, GUS transcriptional fusions were made, with GUS as the reporter gene. Then, using the CRISPRCAS9 method, previously, loss of function mutant lines were generated. Physiological test were made on effect of loss of function of OsHKT1;3 on Na⁺ and K⁺ accumulation in rice tissues during salt stress on different tissues (roots, sheath and leaf blade). The results obtained show that all mutants are homozygous, induce a frame shift and being present at the beginning of the gene (1^st exon). OsHKT1;3::GUS showed strong GUS activity, expressed mainly in vascular tissues and did not show a significant difference in the expression with the NaCland water-treated parts. In contrast, the GUS activity of the OsHKT1;3 promoters in NaCl-treated leaves was greater than that in water-treated leaves. The results of this study show that in wild plants, increasing the Na⁺ concentration in the culture medium has the effect of increasing the Na⁺ content of the tissues generally, the old leaves accumulating more Na⁺ . When the Na⁺ concentration increases, the K⁺ content decreases in roots and old leaves, but varies little in young leaves. Analysis of Na⁺ storage in tissues also shows that Na⁺ levels are higher in the lower parts of the leaf than in the upper parts. Analysis of Na⁺ storage in tissues also shows a Na⁺ in leaf blade more evenly distributed among the first leaves than in WT plants. Taken together, these results suggest that OsHKT1;3 gene plays a role in the accumulation of Na⁺ in old leaves.

6 illus, 16 ref

025519   HARIBALAGANESH R, ROSY J C, ROHITA S, AISHWARYA C, BRINTHA S, RAHUL S, SUNDAR K (Biotechnology Dep, Kalasalingam Univ, Krishnankoil- 626 126, Email: sundarkr@klu.ac.in) : Antiviral drugs against Ebola: A structure based virtual screening approach. Indian J Biotechnol 2018, 17(1), 176-84.

Repression of highly lethal Ebola virus outbreaks poses a serious public health challenge. Ebola haemorrhagic fever is a highly lethal disease for which there are no effective therapeutic or preventative treatments. Hence, there is a necessity to discover new candidate drugs that could have the capability to contain Ebola. Structure based virtual screening method is a method that could accelerate the drug discovery process. In this study, various known antiviral drugs were used as ligands to screen for their binding ability to Ebola proteins by using protein ligand docking. As the crystal structure of the Ebola proteins were not available, these structures were predicted using PS2 or RABTOR X and the predicted structures were validated using Ramachandran plot. Six of the existing drugs exhibited a better binding affinity to various Ebola proteins; they could be used as lead compounds in developing a better drug for controlling Ebola.

11 illus, 5 tables, 23 ref

025518   LASINA K V, PIRAMANAYAGAM S (Bharathiar Univ, Coimbatore- 641 046, Email: lisina777@gmail.com) : Pharmacophore modelling, 3D-QSAR and docking study of HIV inhibitor. Indian J Biotechnol 2018, 17(1), 160-75.

Human immunodeficiency virus (HIV), the causative virus for acquired immunodeficiency syndrome (AIDS) has become the world’s greatest dispute. Inhibition of nucleocapsid protein domain 7 NCp7 receptors have been sturdily pursued as a promising target for the treatment HIV AIDS. A set of 36 thioesters derivatives has been reported as HIV NCp7 inhibitor was analyzed by employing PHASE method to inspect the structural requirements for diverse analogues to inhibit NCp7 receptors and to obtain a highly predictive model used for designing of novel NCp7 receptors inhibitors. A united study of pharmacophore prediction, atom based 3D-quantitative structure–activity relationship (QSAR) and molecular docking approaches were carried out on pyridinioalkanoyl thiolesters derivatives to understand their structural requisites and binding mode of the best fitted ligand for NCp7 inhibitory activity. Five point pharmacophore hypothesis AADDR (two acceptors, two hydrogen donor, one aromatic ring) yielded a statistically significant 3D-QSAR model with partial least square (PLS) factors 5, regression coefficient value of (R²) = 0.9625, cross validation coefficient value of (Q²) = 0.7775, root mean square error (RMSE) = 0.1358. The core structure of nucleocapsid NCp7 domain is docked with the compounds obtained from ZINC and NCI. Docking study also revealed the binding orientation of active ligands at active residues of NCp7. Using pharmacophore based database search, 3D-QSAR and docking studies, we identified ZINC29569253 as a stable inhibitor. The geometry and type of this pharmacophore model give emphasis to important binding features which will be useful for the design of selective HIV NCp7 inhibitors.

10 illus, 5 tables, 16 ref

025517   PAI S R, UPADHYA V, HEDGE H V, JOSHI R K, KHOLKUTE S D (ICMR- National Institute of Traditional Medicine, Belagavi- 590 010, Karnataka, Email: drpaisr@gmail.com) : In vitro rapid multiplication and determination of triterpenoids in callus cultures of Achyranthes aspera Linn. Indian J Biotechnol 2018, 17(1), 151-9.

Achyranthes aspera Linn. was studied for its in vitro rapid multiplication, callus initiation, proliferation and triterpenoid determination. The plant was also evaluated for pretreatment, sterilization and culture initiation during the study. Best results for vigorous cultures were obtained for plants pretreated with 0.2 % (w/v) carbendazime + 3 drops of Tween 20 for 10 min, and double min) solutions. sterilized In using vitro both plantlet sodium regeneration hypochloride in A. (NaOCl) aspera was (4.0 % achieved v/v for from 2 min) axillary and mercuric explants chloride cultured (HgClon Linsmaier 2) (0.1 % w/v & Skoog for 2 (LS) fortified with 6-benzyl amino purine (BAP) alone and/or in combination with Zeatin (Zn), Kinetin (Kn), thidiazuron (TDZ), naphthalene acetic acid (NAA). The combination of BAP (3.00 mg/L) and TDZ (0.10 mg/L) was the best for multiple shoot induction and rapid proliferation. The developed shoots were successfully rooted on medium containing NAA (1.5 mg/L). Rooted plantlets were regenerated and were successfully established in soil with a survival rate of 90 %. The protocol developed for multiplication is rapid and efficient for in vitro propagation of A. aspera. Further, a combination of 2,4-D (3.00 mg/L) and BAP (0.50 mg/L) fortified in LS basal medium, showed significant response with enhanced growth and proliferation of callus. Callus obtained from the 10 combinations were extracted individually using an ultrasonicator, and the extracts were subjected for reverse phase ultra-fast liquid chromatography (RP-UFLC) analysis. Higher amount of betulinic acid (3-hydroxylup- 20(29)-en-(28)-oic acid) and oleanolic acid (3β-hydroxyolean-12-en-28-oic acid) were accumulated on the medium supplemented with 3.00 mg/L of 2, 4-D and 0.50 mg/L of BAP.

3 illus, 4 tables, 41 ref

025516   WANG J, LI X, ZHANG L, ZHANG Y, PENG Y, GUO M, ZHANG R, HUANG J (Minzu Univ of China, Beijing 100 081, People's Republic of China, Email: wangjunli1698@163.com) : Developing micropropagation protocol and analyzing peroxidase activity during morphogenesis in Arisaema decipiens Schott, a medicinal plant. Indian J Biotechnol 2018, 17(1), 145-50.

An efficient micropropagation system for Arisaema decipiens Schott, an ethnic medicinal plant, has been developed. Calluses were induced from petiole explants on Murashige and Skoog (MS) medium supplemented with 0.5 mg l^-1 N⁶-benzyladenine (BA) and 1.0 mg l^-1 2,4-dicholorophenoxyacetic acid (2,4-D). The effect of 1.0 mg l^-1 2,4-D in combination with 1.0 mg l^-1 BA was much more pronounced for the callus proliferation, and fresh weight of callus increased 11.72 times after 5 week of culture. Significantly more protocorm-like bodies (PLBs) could be obtained on calluses exposed to 0.5 mg l^-1 indole-3-acetic acid (IAA) in combination with 0.5 mg l^-1 BA. In this medium, the PLBs transformed into shoots, and frequency of shoot induction was about 82 %. Up to 100 % of the regenerated shoots formed complete plantlets on MS medium supplemented with 0.5 mg l^-1 IAA in combination with 0.5 mg l^-1 BA, with an average of 10.35 roots and 7.43 cm long per shoot. Peroxidase (POD) expressions were measured by polyacrylamide gel electrophoresis during shoot induction and differentiation phases. The results showed that POD activity was connected with the different states of shoot morphogenesis.

2 illus, 3 tables, 20 ref

025515   CHAUHAN R S, JHA S K (Forest Biology and Tree Improvement Dep, Navsari Agricultural Univ, Navsari- 396 450, Gujarat, Email: sumanfort@gmail.com) : Genetic stable mass propagation of Acacia mangium Willd. from mature plus tree. Indian J Biotechnol 2018, 17(1), 128-33.

The study describes an efficient, reproducible and stable in vitro propagation protocol for Acacia mangium from nine years old phenotypically superior plus tree. For effective control of contamination explants were dipped in absolute alcohol for 1 minute followed by 0.1 % mercuric chloride solution for 6 minutes. Maximum establishment (87.8 %) and shoot proliferation was achieved with an average value of 5.7 shoots per explants and shoot length of 2.8 cm in Murashige & Skoog (MS) media supplemented with 6.66 μM 6-benzylaminopurine (BAP) and 0.465 μM kinetin. The addition of 9.80 μM indolebutyric acid (IBA) in half MS media gave maximum rooting (88.3 %) with average 2.7 roots per microshoots with longest root of 18 cm. Rooted plants were hardened and successfully established in the soil. Random amplified polymorphic DNA (RAPD) profile of micropropagated plants shown no change in genetic fidelity up to four growth cycles.

4 illus, 4 tables, 26 ref

025514   SINGH N, MAHAR K S, VERMA S, MEENA B, ROY R K, TEWARI S K, GOEL A K, RANA T S (CSIR-National Botanical Research Institute, Lucknow- 226 001, Email: ranatikam@gmail.com) : Molecular analysis of genetic variability and relationship among Gladiolus cultivars. Indian J Biotechnol 2018, 17(1), 118-27.

In the present study, we analyzed genetic variability and relatedness in 62 Gladiolus cultivars using directed amplification of minisatellite DNA (DAMD) and inter simple sequence repeats (ISSR). The maximum inter-cultivar average genetic distance was 0.46 between Tiger Flame and Snow Flower cultivars, while the corresponding least genetic distance 0.14 was between Friendship Pink and Friendship White cultivars, respectively. The cumulative analysis carried out for the data generated with DAMD and ISSR markers showed 83.32 % polymorphism across all the Gladiolus cultivars. This level of polymorphism resulted in the present investigation revealed that the amount of genetic variability in the Gladiolus genome is relatively high. The Jaccard’s similarity coefficient and clustering of genotypes in the unweighted pair group method with arithmetic mean (UPGMA) dendrogram revealed that some of the cultivars are closely related whereas others were found quite distinct from each other. The UPGMA dendrogram resulted in the identification of five major clusters. The present study further demonstrates that DAMD and ISSR are useful markers to elucidate the genetic variability and relationships amongst Gladiolus cultivars, and is a prelude for further utilization of promising and genetically divergent materials in the breeding programmes.

3 illus, 6 tables, 39 ref

025513   GAYACHARAN, BISHT I S, PANDEY A, YADAV M C, SINGH A K, PANDRAVADA S R, RANA J C (ICAR-NBPGR Regional Station, Nainital- 263 132, Email: bishtis@rediffmail.com) : Population structure of some indigenous aromatic rice (Oryza sativa L.) landraces of India. Indian J Biotechnol 2018, 17(1), 110-7.

The Indian sub-continent is home to a large number of indigenous aromatic landraces, which may serve as valuable genetic resources for future quality rice improvement to meet the ever growing demand for quality rice. In the era of high yielding modern varieties there is an urgent need to conserve and mainstream in production systems the valuable aromatic landraces occurring in traditional subsistence farming areas across the country. Understanding the population structure and adaptive variations of landraces is considered useful in this regard. In order to explore the genetic structure and diversity, 256 individuals of 8 aromatic landrace populations from four eco-geographic regions were genotyped using 17 SSR markers in the present study. Sixty-nine alleles were recorded and none of them were found common across all landraces. Twenty- six alleles were found unique to an individual landrace population. Kala Joha from Assam was most diverse in terms of effective number of alleles, expected heterozygosity and Shannon’s information index. Analysis of molecular variance (AMOVA) revealed maximum variation of 39.36 % among populations within groups followed by among groups (30.52 %) and the rest (30.12 %) within populations.

3 illus, 4 tables, 31 ref

025512   SABARA P, VAKHARIA D (Biotechnology Dep, Junagadh Agricultural Univ, Junagadh- 362 001, Email: priteshrhsabara@gmail.com) : Genetic diversity study in papaya (Carica papaya L.) cultivars using RAPD and ISSR markers. Indian J Biotechnol 2018, 17(1), 101-9.

Thirteen papaya cultivars and lines were used to assess genetic diversity through dominant PCR based markers (16 RAPD and 12 ISSR primers). RAPD (random amplified polymorphic DNA) primers gave 58 polymorphic bands out of 126 with 47.19 % polymorphism and ISSR (Inter simple sequence repeats) primers produced 49.80 % polymorphism with 37 polymorphic bands out of 65 bands. Dendrogram showed clear grouping ancestrally related papaya cultivars. Principle coordinate analysis (PCoA) showed congruence with dendrogram pattern while bootstrap values at major nodes in both marker systems as well as in pooled data indicated robustness of cluster pattern in dendrogram construction. Analysis of molecular variance (AMOVA) indicated that highly significant genetic variability was obtained through both marker systems while cultivars showed noticeable variation in case of RAPD markers. Mantel’s test between similarity and cophenetic coefficient matrices in different combination showed poor to very good correlation between matrices. This study indicates relation between papaya cultivars based on their ancestral relationship which can be utilized for conservation of important cultivars and development of future strategies for crop improvement programme.

2 illus, 5 tables, 28 ref

025511   BORA L, SINGH A K, KUMAR A, METWAL M (Agriculture Coll, Nainital- 263 139, Email: lokeshbora36099@gmail.com) : Morphological and microsatellite marker based polymorphic assessment of genetic diversity and relationship of mango (Mangifera indica L.). Indian J Biotechnol 2018, 17(1), 91-100.

Genetic diversity of 19 genotypes of mango was characterized both by morphological and 20 simple sequence repeat (SSR) markers. Characterization of mango genotypes based on morphological and molecular basis is a better approach for designing breeding projects. On the basis of the present findings it was observed that “Sabri” and “Amrapali” showed dwarf stature, while “Swarna Jahangir” was found to be vigorous in its growth. The unweighted pair group method of arithematic- average (UPGMA) dendrogram based on genetic distance segregated the 19 mango genotypes into two main clusters. The polymorphic information content (PIC) values ranged from 0.38 to 0.81. Jaccard’s similarity coefficient values ranged from 0.15 to 0.79 with polymorphism of 77.5 per cent. Three unique fingerprints were identified in three genotypes which can help in varietal identification. A total of 49 loci (42 polymorphic and 7 monomorphic) were detected with amplified size range of 110 to 359 bp. The maximum numbers of loci (i.e. 5) were detected by the primer MiSHRS-48. Out of 20 SSR primers, 18 were polymorphic. Pusa Surya was found to be the most diverse genotype both morphologically and genetically. The similarity for Pusa Surya was 15 per cent with remaining Indian genotypes. The significant variation exists among the genotypes based on morphological and biochemical characters but with the use of SSR markers, assessment of the genetic diversity can help us to plan a future breeding programme using the diverse parent.

3 illus, 5 tables, 58 ref

025510   ATRI C, SHARMA S, BANGA S S (Plant Breeding and Genetics Dep, Punjab Agricultural Univ, Ludhiana- 141 004, Email: chhayaatri@pau.edu) : Genome specific microsatellites in wild crucifers: Cross species/genera transferability. Indian J Biotechnol 2018, 17(1), 80-90.

A high degree of colinearity is now known to exist between closely related species in Brassicaceae, which theoretically allows the exchange of markers between them. Objective of this study was to explore the transferability of simple sequence repeat (SSR) primer pairs identified in crop Brassicas to related wild crucifers. Here, we report and validate transferability of 92, 67 and 105 SSR primer pairs, identified in three diploid Brassica genomes, to Brassica fruticulosa, Erucastrum abyssinicum and Diplotaxis tennuisiliquae with respective amplification of 259, 141 and 291 alleles. One thousand six hundred and thirty three (394 in RL 1359, 451 in PBR 210, 402 in RLC 1 and 386 in UP) alleles were detected in cultivated species. In cultivated Brassica species, the average number of alleles amplified were 3.31, 2.19, 2.97 whereas in wild species the values recorded were 2.70, 1.67, 2.32 for A, B and C genome SSR markers, respectively, however respective average allele values per germplasm set were 2.82 (cultivated) and 2.23 (wild). The average polymorphism information content (PIC) values of A genome primer pairs in wild species was 0.77, ranging from 0.32 to 0.94. These transferable markers can now be exploited for further genetic and introgressive breeding studies. The transferability success generally decreased as the evolutionary distance between the source and target species increased.

3 illus, 2 tables, 27 ref

025509   SINGH A, SHARMA J K (Seed Science and Technology Dep, C S K Himachal Pradesh Krishi Vishwavidyalaya, Palampur- 176 062, Email: singhadr@yahoo.com) : Molecular characterization of parental lines and hybrids of maize cultivated in north-western Himalayan region using microsatellite markers. Indian J Biotechnol 2018, 17(1), 74-9.

Maize is an important cereal crop of north-western Himalayan region of India with wide ranging and diversified uses which is resulting in the release and recommendation of new hybrids of maize for the region. The use of parental lines with a narrow genetic base by various private and public sector seed agencies may cause a risk of genetic diversity loss. Molecular characterization of hybrids and parental lines by DNA fingerprinting provides knowledge of the genetic relationship among them thus preventing the risk of increasing uniformity. The technique is helpful for molecular identification and hybrid purity testing. The present study was undertaken to carry out microsatellite marker based DNA fingerprinting of ten maize hybrids and their parental lines for molecular identification and assessment of genetic diversity. Twenty three microsatellite markers were found to be polymorphic. Number of alleles per locus ranged from 2 to 6 with a mean of 3.3. Mean expected heterozygosity and polymorphic information content was 0.58 and 0.52, respectively. Average Jaccard’s similarity coefficient observed was 0.46 revealing an average genetic diversity of 54 % among various genotypes studied. Cluster analysis delineated the maize hybrids and parental lines into four main clusters, A, B, C and D at 0.32 similarity coefficient.

2 illus, 2 tables, 20 ref

025508   KUMAR S, SOHU V S, SINGH R P, GUPTA S K, SRIVASTAVA P, BAINS N S (Plant Breeding & Genetics Dep, Punjab Agricultural Univ, Ludhiana- 141 001, Email: kumarsatish227@gmail.com) : Investigating the role of high molecular weight glutenin subunits (HMW-GS) protein in end use quality of Indian flat breads. Indian J Biotechnol 2018, 17(1), 65-73.

Indian unleavened flat breads more commonly known as chapati are core to existence for two-third of its population. Glutenins and gliadins constitute gluten which gives extensibility and elasticity to the dough, traits which are of great importance in bread making. However, limited studies have been attempted to explore the relationship and functionality of the protein subunits with the quality of the unleavened flat breads. Chapati characterization and molecular analysis was carried on two set of genotypes firstly, different commercial wheat cultivars and genetic stocks for various quality parameters, secondly three back cross derived recombinant populations from the parents with different HMW-GS at Glu 1B locus to associate chapati quality with the glutenin subunits. Significant variation obtained in the genotypes revealed that tall varieties are distinct in quality followed by derivatives of C 306 and C 591 (7.8 for both DI 9 and DI 105). The SDS-PAGE analysis of the high molecular weight glutenin protein revealed that the subunit ‘20’ at Glu1B locus is unique to tall traditional wheats. However, such investigations in the recombinant populations indicated complex inheritance of this trait.

2 illus, 5 tables, 15 ref

025507   MYTHILI J B, CHETHANA B S, RAJEEV P R, GANESHAN G (Biotechnology and Plant pathology Dep, Indian Institute of Horticultural Research, Bangalore- 560089, Email: jbm@iihr.ernet.in) : Chitinase gene construct from Trichoderma harzianum proved effective against onion purple blotch caused by Alternaria porri. Indian J Biotechnol 2018, 17, 50-6.

Purple blotch caused by Alternaria porri (Ellis) Ciffis the most devastating disease of onion prevalent in different parts of the country. In the absence of a resistant variety in the gene pool, gene transfer technique becomes appropriate as an alternative tool for genetic improvement. Accordingly, chitinase gene especially from the biocontrol agent Trichoderma harzianum Rifai which is used against several fungal pathogens was selected as the candidate gene. Onion transformation is difficult and Allium species are recalcitrant to transformation. Hence, before using this gene for transforming onion for conferring resistance/tolerance to Alternaria porri, chitinase gene was validated against purple blotch pathogen in a model plant system, tobacco. Tobacco transformants with T. harzianum chitinase (Th-chit) gene under the control of a strong constitutive CaMV 35S promoter with NPT-II selection marker were generated. The presence and expression of the transgene was confirmed through PCR and RT-PCR analysis, respectively. Bioassay of primary transgenic plants against A. porri through in vitro detached leaf bioassay revealed significant reduction in lesion size ranging from 73 - 100 % as well as mycelial inhibition to the extent of 25 - 65 % over the control (untransformed) plants. The results suggest that chitinase gene from T. harzianum can be used as a candidate gene for conferring resistance to A. porri in onion.

8 illus, 26 ref

025506   JOSE A A, ANUSREE G, PANDEY A, BINOD P, PANDEY A, BINOD P (CSIR-National Institute for Interdisciplinary Science and Technology, Thiruvananthapuram- 695 019, Email: binodkannur@niist.res.in) : Production optimization of poly-γ-glutamic acid by Bacillus amyloliquefaciens under solid-state fermentation using soy hull as substrate. Indian J Biotechnol 2018, 17(1), 44-9.

Poly-γ-glutamic acid (PGA) is a biodegradable polymer with wide applications in the field of medicine, cosmetics, food and agriculture. The aim of the present work was to develop a microbial method to produce PGA under solid-state fermentation. Soy hull was used as the solid media in fermentation by Bacillus amyloliquefaciens. Central composite rotary designs (CCRD) was used to optimize the production of PGA and also studied the pattern of mutual interactions between the variables. The values of P < 0.05 indicated that the model terms were significant with the PGA yield of 111.2 mg/gds at initial moisture 75 %, incubation time 60 h and inoculum size 4.7 ml. Characterization of the purified PGA was done by gel permeation chromatography and fourier transformed infrared (FT-IR) spectroscopy. The choice of soy hull as substrate and the process parameters optimization by response surface methodology (RSM) were found to be significant in PGA production. Due to low-cost nature of the process, this study shows the possibility for establishing large scale production of PGA in cost-effective manner.

4 illus, 4 tables, 22 ref

025505   RATHINASAMY P, THAYUMANAVAN P (Public Health Dep, Aksum Univ, Ethiopia, Email: sami7bio@gmail.com) : Identification of efficient dye decolorizing laccase producing fungi from Kolli Hills. Indian J Biotechnol 2018, 17(1), 33-43.

Laccase is one of the most promising ligninolytic enzymes for the industrial application and ecofriendly bioremediation process. Twenty-five carpophores were collected from different places of Kolli Hills (Namakkal district Tamilnadu, India) and screened on the solid media containing guaiacol, which enabled the detection of laccase secretion. Three positive strains were isolated and the quantitative production of laccase was determined in submerged culture to select hypersecretory strain for further study. Among the three strains, “ST02” the best producer of laccase was selected and was analyzed for the dye decolorization potential using dyes like Poly R-478 and Remazol Brilliant Blue R (RBBR). Identification of the isolated organism was carried out by classical and molecular methods. Approximately 625 bp of the ST02 5.8S rDNA was amplified by polymerase chain reaction (PCR). The phylogenetic relationship of the isolated strain was studied by comparing the internal transcribed spacer (ITS) sequences of ST02 with similar related sequences deposited in the GenBank database. The present study showed that relatively simple plate test screening method and ITS analysis can be used for identification of laccase producing new strain. The isolated organism was designated as Pleurotus ostreatus IMI 395545.

7 illus, 1 table, 48 ref

025504   VIGNESH V, SATHIYANARAYANAN G, PARTHIBAN K, KUMAR K S, THIRUMUGAN R (Animal Science Dep, Bharathidasan Univ, Tiruchirappalli- 620 024, Email: ramthiru72@gmail.com) : Functional assessment of subtilosin A against Aeromonas spp. causing gastroenteritis and hemorrhagic septicaemia. Indian J Biotechnol 2018, 17(1), 27-32.

Anti-Aeromonas and cell membrane lytic bacteriocin substance, subtilosin A producing Bacillus subtilis VT03 was explored. Strain VT03 was isolated from freshwater fish (Tilapia) intestine and screened for its antimicrobial activity against four pathogenic strains of Aeromonas spp. causing gastroenteritis and hemorrhagic septicaemia. Isolate (VT03) was identified showing inhibition in agar spot assay. The strain VT03 was the one exhibiting strong inhibition and identified as Bacillus subtilis using 16S rRNA sequencing. Cell free supernatant (CFS) of the strain VT03 was active against pathogenic strains of Aeromonas spp, subsequently CFS was partially purified and designated as PPB-VT03 showing inhibition against A. hydrophila ATCC 49140. PPB-VT03 completely lost its activity upon treating with proteinase K revealing that the defense molecule could be proteinaceous in nature. Based on polymerase chain reaction (PCR), functional gene coding for subtilosin A (sboA) was found to be present whereas subtilin (spaS) was absent. The role of partially purified bacteriocin of isolate VT03 (PPB-VT03) through FTIR and SEM analysis revealed the activity of cell lysis. The study demonstrated the potential use of subtilosin A producing Bacillus subtilis as a potent source for antibacterial peptide.

3 illus, 34 ref

025503   BAKHSHI M, EBRAHIMI F, NAZARIAN S, ZARGAN J (Biology Dep, Imam Hossein Univ, Tehran, Iran, Email: kpnazari@ihu.ac.ir) : Computational analysis and gene cloning: Design and preparation of a multi subunit vaccine consisting of EspA, Stx2B and Intimin antigens against enterohaemrrhagic Escherichia coli. Indian J Biotechnol 2018, 17(1), 16-26.

Escherichia coli O157:H7 is an intestinal pathogen that made diarrhoea, haemolytic uremia syndrome (HUS) and hemorrhagic colitis (HC) in patients. Roles of EspA and Intimin at the beginning of bacteria colonization in intestine are critical. Destruction of protein synthesis route with shiga toxins of E. coli O157:H7 is mediated through B-subunit of toxins. In this study, in silico approaches were performed to design a suitable construct from EspA, Intimin and Stx2B and a recombinant chimeric antigen was produced. Bioinformatics analyses such as physicochemical data, mRNA folding, 3D structures of chimera and various immunoinformatic data, such as linear and conformational B-cell epitopes, T-cell epitopes were reported according to authentic data base. The chimeric gene was prepared as synthetic construct after designing and cloning. The validation result showed that 83.9 % residues lie in favoured or additional allowed region of the Ramachandran plot. Epitope prediction results proved very good distribution of conformational B-cell epitopes in the 3D structure of chimera. The identified T-cell epitopes are apt to bind MHC molecules. A good quantity of recombinant chimeric antigen was achieved in host cells. From in silico approach, an appropriate multi subunit vaccine candidate was designed and prepared for immunological examinations.

9 illus, 5 tables, 35 ref

025502   GANESHPURKAR A, SALUJA A (Gujarat Technological Univ, Ahmedabad- 382 424, Email: akspharmacy@yahoo.com) : In silico interaction of rutin with some immunomodulatory targets: A docking analysis. Indian J Biochem Biophys 2018, 55(2), 88-94.

The use of plant products as immunomodulator has a long history. Forms eternal era, plant, mineral and animal products are used as drugs for the treatment of various diseases. The present-day synthetic compounds find their leads in natural products. The process of immunomodulation retunes the immune system of an individual by restoring routine functions. Research on immunomodulators from natural sources has been extensively studied for modulation of immune system along with protection and prevention of diseases. Rutin, a flavonoid has been investigated for its potential immunomodulatory effects. Increased particle clearance, decreased inflammation, increased leucocyte and antibody production was observed with administration of rutin in rats. The present study is focused on exploring in silico interaction of rutin with some chemokines and inflammatory targets. In this study, rutin was docked with TNF-α, IL-1β, IL-6, and NOs. Docking studies revealed the excellent interaction of rutin with these targets. The result of present study provides insight for the discovery of novel molecules for immunomodulation and treatment of inflammatory disorders. Findings from the present study show that rutin may interact with several chemokines and inflammatory mediators. Further studies on rutin and associated flavonoids are necessary to develop and establish QSAR and QSPR studies which may serve a stepping stone for the development of novel and safe immunomodulator. Rutin therefore, can be considered for development of an immunomodulatory agent.

4 illus, 35 ref

025532   WAGH S S, SAHOO N R, RAWAT C, VERMA A D (ICAR- Indian Veterinary Research Institute, UP- 243 122, Email: vet.nihar@gmail.com) : Jejunal expression of Mucin4 gene in pigs differentially adhesive to diarrhoeagenic E. coli. Environ Ecol 2018, 36(2), 443-5.

The E.coli mediated piglet diarrhoea is the major cause of the piggery industry which causes severe economic hardship to piggery sector. E. coli binds to the brush border of the epithelial cells of the intestine on this basis of receptors and fimbrae adhesion pattern. Among the putative candidate genes associated with adhesion pattern, mucin4 gene was localized on targeted region of SSC13 and considered as a putative candidate gene due to its biochemical property as well as physical location. The presentinvestigation was conducted to study mucin4 gene expression profile in different adhesion phenotypes. A total of 4 types of adhesion pattern were observed with different frequencies. RT-PCR analysis revealed that mucin4 mRNA expression level was different across the differentially adhesive phenotypes.

1 illus, 1 table, 6 ref

025530   SHARMA M R, KUMARI S, MISHRA M (ICAR- Central Institute of Subtropical Horticulture, Lucknow- 226 101, Email: manasiibsindia@gmail.com) : Micropropagation of wilt resistant, inter-specific (Psidium molle x Psidium guajava) rootstock of guava. Plant Arch 2018, 18(1), 1170-4.

The present study describes a protocol for in vitro micropropagation of wilt resistant, hybrid rootstock of guava, developed at ICAR- Central Institute for Subtropical Horticulture, Lucknow April to June was found to be the most suitable season for collection of explants. A combination of 0.1 % Carbendazim and 0.05 % Metalaxyl for pre-washing followed by surface sterilization with 0.1 % Mercuric chloride for 7 minutes was found effective in controlling contamination. In order to control oxidative browning, applying wax to the cut ends of the explants yielded better results than the use of antioxidants. Out of all the different media preparations used, MS + 0.1 mg/L IAA+ 4 mg/L BAP+ 30g sucrose gave maximum proliferability (2.6 shoots/explant) and maximum leaf number/shoot (5.6) in the shortest duration viz. 7 days. The regenerated micro-shoots were transferred on rooting media containing MS + 2 mg/L IBA. The rooted plants were acclimatized on sterilized coco-peat supplemented with MS salt solution.

3 illus, 2 tables, 40 ref

025529   AL_AMIRY A H A (Horticulture Dep, Basrah Univ, Iraq, Email: aqeelhadi6@gmail.com) : Comparison between three different methods for isolation genomic DNA from dry date palm leaves without liquid nitrogen. Plant Arch 2018, 18(1), 1011-4.

Three different methods for Isolation genomic DNA from Date palm leaves is described. The leaves was dried and ground before DNA extraction by CTAB development method [100 mM Tris- HCl (pH 8.0), 20 mM EDTA , 4M NaCl, 6 % CTAB, 2 % Polyvinylpyrrolidone, 0.2 % B-mercaptoethanol and 15 µl of proteinase K] and compared with two other methods, the results shows the developing method gives highly quality of DNA (A_260/280 = 1.6 to 1.9) in all three Date palm cultivars under tests, this study found the development method is very suitable to DNA extraction from dried leaves without liquid nitrogen.

3 illus, 16 ref

025528   RATHI M H, MOLAN A, ISMAIL M H (Biotechnology Dep, Diyala Univ, Diyala, Iraq) : The impact of the extracting solvent and the cultivar on the determination of total phenolic contents and anti-radical activities of extracts from roasted date seeds of two date cultivars cultivated in Iraq. Plant Arch 2018, 18(1), 830-4.

The objectives of the current study were to determine the total phenolic content (TPC) and the anti-radical activities of extracts prepared from roasted date-seeds of two date cultivars (Zahdi and Barhi) using different solvents. The TPC was measured using Folin–Ciocalteau method; while the antioxidant activity was measured using DPPH-radical scavenging assay. The results showed that the extracting had a significant role in the determination of the total amount of the TPC and the free radical scavenging activity of roasted date-seeds. Acidification of the water with hydrochloric acid (1% and 5%) resulted in a significant increase (P< 0.001) in both the TPC and anti-radical activity in comparison with the distilled water alone. Extracts prepared from the powdered roasted seeds of Zahdi cultivar had significantly higher (P< 0.5) TPC and freeradical scavenging activity than their counterparts from Barhi cultivar and in all solvents used. In the extracts from the powdered roasted seeds of both cultivars, positive correlation was found between the TPC and the anti-radical activity, indicating that the phenolic compounds are the main ingredients contributing to the free-radical scavenging activity of the two Iraqi date cultivars. In conclusion, the solvent used for the extraction and the cultivar of the date play a significant role in determining the TPC and free radical scavenging activity and that the roasted date-seeds powder could be a good potential source of bioactive compounds that have anti-radical capacities.

2 illus, 1 table, 26 ref

025527   SATAR J A, AL-THWANI A N, TOTHLI K J (Institute of Genetic Engineering and Biotechnology, Iraq, Email: jinan_2014@yahoo.com) : Detection of Clostridium bolteae in samples of Iraqi autistic children using traditional and molecular methods. Plant Arch 2018, 18(1), 781-4.

Clostridia are widely distributed in natural environments and are inhabitants of both human and animal gastrointestinal tract. These organisms are important pathogens that may cause pseudomembranous colitis, necrotizing enteritis, food poisoning and the other intestinal disorders, such as diarrhea. The objective of this study was to detect C. bolteae from samples of Iraqi autistic children. Stool samples were collected from children under ten years. The samples were subjected to culturing and DNA extraction. Three isolates were detected by traditional method and showed positive results, while in conventional PCR furthermore detection of target bacteria using RT-PCR that is more sensitive, more reliable and fast.

4 illus, 3 tables, 18 ref

025526   SAHAY S, GUPTA M (Biotechnology Dep, Jamia Millia Islamia, New Delhi- 25, Email: meetu_gpt@yahoo.com) : Effect of nitric oxide and abscisic acid on growth determinants in Brassica juncea cultivars subjected to PEG-induced water stress. Plant Arch 2018, 18(1), 737-44.

The aim of the present study was to evaluate the effect of different concentrations of sodium nitoprusside [SNP (10, 50, 100, 150 and 200 μΜ)], and abscisic acid [ABA (0.1, 1, 10, 50 and 100 μM) in absence (-) and presence (+) of polyethylene glycol 6000 [PEG 6000 (10 %)]-induced water stress on growth determinants, in 7 days old hydroponically grown two Brassica juncea cultivars (Pusa Jagannath and Varuna), subjected to 96 hrs exposure. Results revealed that the growth determinants viz., shoot-root length, shoot-root fresh and dry weight and other root development related parameters decreased under PEG, as compared to control. The effect of SNP and ABA with or without PEG was concentration dependent. In particular, SNP 100 µM and ABA 10 µM treatments were optimum, and had efficient effect on promoting the maximum growth determinants of both Brassica cultivars in absence (-PEG) as well as in the presence of (+PEG) of water stress. However, the application of SNP 150 μM, SNP 200 μM, ABA 50 μM and ABA 100 μM had no obvious alleviating effect on PEG toxicity, as well as could not enhance growth in both cultivars when applied even alone.

6 illus, 1 table, 20 ref

025525   AL-FADHAL F A, AL-ABEDY A N, AL-JANABI M M (Plant Protection Dep, Kufa Univ, Iran, Email: fadhl.alfadhl@uokufa.edu.iq) : Molecular identification of novel isolates of Rhizoctonia solani k?hn and Fusarium spp. (matsushima) isolated from petunia plants (petunia hybrida l.). Plant Arch 2018, 18(1), 703-11.

This study was conducted in the Plant Virology Laboratory, College of Agriculture, Karbala University with the aim of isolating and identifying some fungi, isolated from petunia plant (Petunia hybrida L.) roots collected from some private nurseries in Najaf province. The polymerase chain reaction (PCR) technique was used to determine the nucleotide sequence of PCR-amplified products. The results from the PCR amplification and analysis of the nucleotide sequences of PCR amplified products by using BLAST program (Basic Local Alignment Search Tool) showed that the isolated fungi are belonged to Rhizoctonia solani, Fusarium verticilliodes, Fusarium proliferatum, Fusarium solani and Trichoderma harzianum. By comparing the nucleotide sequences generated from PCR products amplified from the ITS1-ITS4 region of the above-mentioned fungal isolates with the nucleotide sequences, belonged to the same fungi and available in the National Center for Biotechnology Information (NCBI), it was found that the identified fungal isolates of R. solani, F. verticilliodes, F. proliferatum were not previously recorded in NCBl, therefore; the identified sequences of R. solani, F. verticilliodes, F. proliferatum have been deposited and recorded in GenBank database (NCBI) under the accession numbers : KX828175, KX828174 and KX828173.

6 illus, 5 tables, 20 ref

025524   PRASAD S C, KISKU A V, SARIN N B (School of Life Sciences, Jawaharlal Nehru Univ, New Delhi - 110 067, Email: neerasarin@rediffmail.com) : Understanding the gamma-vacuolar processing enzyme gene regulation by promoter-gus fusion approach. Plant Arch 2018, 18(1), 679-89.

Vacuolar processing enzymes (VPEs) are vacuole localizing cysteine proteinase, responsible for the maturation of seed storage proteins in plants and are also known to be involved in programmed cell death. In the present study, an attempt has made to understand the regulatory mechanism of AtγVPE gene expression by studying promoter region of AtγVPE. We isolated approximately 1.2 kb fragment upstream of the initiation codon of γVPE gene sequence and deletion mutations were made from 5’ end of promoter region, fused with GUS reporter gene to analyze the transient GUS activities in Nicotiana benthamiana leaves. The full-length promoter (1282 bp) was found to show lower GUS activity in comparison to the deletion fragments. However, P6 (-349 bp) promoter had the maximum activity and on further deletion the activity reduced, with little or no activity in P8 (-176 bp), signifying P6 having the maximal promoter activity. In-silico analysis revealed the presence of several putative cis-regulatory elements in the promoter region, including hormone-responsive elements for ABA, GA, SA and ethylene, elements involved in abiotic stress, development, and several repressor elements. Functional validation of putative γVPE promoter in tobacco leaves also proved the promoter activity was induced by abiotic stress and phytohormones.

3 illus, 1 table, 32 ref

025523   NAIDU M A, PRASAD Y R (Biotechnology Dep, Acharya Nagarjuna Univ, Nagarjunanagar- 522 510, Email: manaidupharmacy@gmail.com) : Synthesis of novel diarylsulfonylureachalcone hybrid molecules with potential in vitro antimicrobial activity. Asian J Pharm 2018, 12 (2), 88-93.

To study the antimicrobial effect of novel diarylsulfonylurea-chalcone hybrids with significant associated morbidity and mortality which is mainly due to the development of microbial resistance to the existing antimicrobial agents. The antimicrobial effect of novel diarylsulfonylurea-chalcone hybrid molecules was evaluated by agar well diffusion method against various strains of bacteria and fungi. Most of the compounds showed promising antibacterial and antifungal activity. The results in the present study suggest that novel diarylsulfonylurea-chalcone hybrids can be used in treating diseases caused by the tested organisms.

3 tables, 11 ref

025522   SAHNOUN M, SAIBI W, BRINI F, BEJAR S (Centre of Biotechnology of Sfax, Sfax Univ, Sfax, Tunisia, Email: mouna.sahnoun@yahoo.fr) : Apigenin isolated from A. americana encodes Human and Aspergillus oryzae S2 α-amylase inhibitions: credible approach for antifungal and antidiabetic therapies. J Food Sci Technol 2018, 55(4), 1489- 98.

Agave americana extract was analyzed by reverse phase HPLC for characterization. Among phenolic compounds identified, apigenin was observed to be present. The finding showed an inhibitory effect of apigenin towards Human and Aspergillus oryzae S2 α-amylases. Apigenin inhibition towards Human and A. oryzae α-amylase activities was observed to be competitive. IC₅₀ and % inhibition of apigenin for A. oryzae α-amylase were 3.98 and 1.65 fold higher than for Human α-amylase. The inhibition of the described biocatalyst activity was significantly lowered when apigenin was pre-incubated with starch. In addition to the catalytic residues, 44 amino acid residues were involved on A. oryzae α-amylase-apigenin interactions while only 11 amino acid residues were exposed for Human α-amylase-apigenin complex. The binding site of apigenin showed 76 polar contacts for A. oryzae S2 α-amylase against 44 interactions for Human α-amylase. The docking studies confirmed the mode of action of apigenin and strongly suggested a higher inhibitory activity towards fungal amylase which was experimentally exhibited. These findings provided a rational reason to establish apigenin capability as a therapeutic target for postprandial hyperglycaemia modulation and antifungal therapy.

4 illus, 2 tables, 29 ref

025521   ALUGOJU P, PERIYASEMY L, DYAVAIAH M (Biochemistry and Molecular Biology Dep, Pondicherry Univ, Pondicherry- 605 014, Email: madhud14@yahoo.co.in) : Quercetin enhances stress resistance in Saccharomyces cerevisiae tel1 mutant cells to different stressors. J Food Sci Technol 2018, 55(4), 1455- 66.

The Saccharomyces cerevisiae TEL1 gene is an ortholog of the human ATM (Ataxia telangiectasia mutated) gene. S. cerevisiae tel1 mutant (tel1∆) lacking Tel1p, share some of the cellular defects with ATM mutation that includes prevention of oxidative damage repair, premature aging and apoptosis. In the present study, we investigated the protective effects of quercetin on the sensitivity of yeast S. cerevisiae tel1∆ cells exposed to oxidative, apoptotic and DNA damaging stress and viability of tel1∆ cells during chronological aging. Quercetin improved the stress resistance of tel1∆ cells when challenged with oxidants such as hydrogen peroxide (H₂O₂), menadine bisulphite (MBS) and tertiary butyl hydroperoxide (t-BHP) by scavenging reactive oxygen species (ROS). Quercetin protected the tel1∆ cells from acetic acid-induced apoptotic cell death and sensitivity against hydroxyurea. We found that quercetin attenuated ROS accumulation and apoptotic markers in tel1∆ cells and therefore an increase in cell viability during chronological aging. Our results from the S. cerevisiae model, suggest that use of quercetin as a food supplement might alleviate oxidative stress mediated DNA damage, apoptosis and age related damaging effects in AT patients and also improve health beneficial effects in humans.

4 illus, 35 ref

025520   ABBASILIASI S, TAN J S, IBRAHIM T A T, RAMANAN R N, KADKHODAEI S, MUSTAFA S, ARIFF A B (Bioprocess Technology Dep, Univ Putra Malaysia, Selangor, Malaysia, Email: arbarif@upm.edu.my) : Kinetic modeling of bacteriocin-like inhibitory substance secretion by Pediococcus acidilactici Kp10 and its stability in food manufacturing conditions. J Food Sci Technol 2018, 55(4), 1270- 84.

This paper deliberates the modelling and validation of bacteriocin-like inhibitory substance (BLIS) secretion by Pediococcus acidilactici Kp10 at different agitation speeds in a stirred tank bioreactor. A range of models namely the re-parameterised logistic, Luedeking-Piret and maintenance energy were assessed to predict the culture performance of the said bacterium. Growth of P. acidilactici Kp10 was enhanced with increased agitation speed up to 600 rpm while BLIS secretion was maximumat 400 rpm but decreased at higher agitation speed. Growth of P. acidilactici aptly subscribed to the re-parameterised logistic model while BLIS secretion and lactose consumption fitted well with the Luedeking-Piret model. The models revealed a relationship between growth of the bacterium and BLIS secretion. Bacterial growth and BLIS secretion were largely affected by the agitation speed of the stirred tank bioreactor which regulated the oxygen transfer to the culture. BLIS secretion by P. acidilactici Kp10 was however enhanced in oxygen-limited culture. The study also assessed BLIS from the perspective of its stability when subjected to factors such as temperature, pH and detergents. Results showed that BLIS produced by this strain was not affected by heat (at 25–100 °C for 20 min and at 121 °C for 15 min), surfactant (Tween 40, 60 and 80 and urea), detergents (up to 1 % SDS), organic solvents (50 % each of acetone, methanol and ethanol) and stable ina wide range of pH (2–10). The above information are pertinent with reference to commercial applications of this bacterial product in food manufacturing which invariably involve various sterilization processes and subjected to a wide pH range.

2 illus, 4 tables, 64 ref

025501   AKHTER M Z, REJESWARI M R (Biochemistry Dep, All India Institute of Medical Sciences, New Delhi- 110 029, Email: rajeswari3011@hotmail.com) : Anticancer activity of HMGA1 promoter targeting triplex forming oligonucleotide in HeLa cell line. Indian J Exp Biol 2018, 56(3), 219- 29.

High mobility group protein A1 (HMGA1) acts as an architectural transcription factor and regulates transcription of various genes. Upregulation of HMGA1 has been described in a large number of human malignancies and serves as a ‘tumor marker’. Due to its role in neoplastic transformation and tumor progression, hmga1 is considered as a promising therapeutic target. In the present study, we investigated the interaction of triplex forming oligonucleotide (TFO) of 18 bps targeted to hmga1 promoter (-1917 to -1940) and its influence on the expression of HMGA1 in HeLa cells. Stability of DNA triplex was characterized using various biophysical and thermodynamic methods and was confirmed by gel retardation assay using γ-³²P [ATP]. Treatment of HeLa cells with hmga1 specific TFO significantly downregulated HMGA1 expression at mRNA, protein levels (~48 %) and inhibited cell proliferation as investigated by RT-PCR, Western blot and Flow cytometry. The findings of the study suggest that TFO-mediated inhibition of hmga1 expression can be a promising strategy for modulation of gene expression and for inhibition of cancer cell proliferation. Moreover, DNA triplex-based therapeutic approaches hold promise in combating cancers associated with HMGA1 overexpression.

9 illus, 45 ref

025500   SINDHUJAA V, GNANARAJ M, VIJI M, KARUPPANAPANDIAN T, MANOHARAN K (Plant Morphology and Algology Dep, Madurai Kamaraj Univ, Madurai- 625 021, Email: manoharank1956@gmail.com) : Induction of high frequency somatic embryogenesis and analysis of developmental stagewise expression of SERK1 gene during somatic embryogenesis in cultures of Vigna radiata (L.) R.Wilczek. Indian J Exp Biol 2018, 56(3), 180- 93.

Vigna radiata (L.) R.Wilczek (Fabaceae), commonly called Green gram or Mung bean, is an important legume with potential nutritional, medicinal and health benefits cultivated widespread throughout the rain-fed areas of arid and semi-arid tropics and subtropics. Being an affordable source of carbohydrate, vitamins, minerals and phytonutrients besides protein, Green gram finds demand for its nutrient digestibility, food processing properties and bioavailability. Though India ranks top in world mung bean production (>50 %), it is unable to meet the local demand. Biotic and abiotic stresses restrict mung bean yield considerably and researchers have been working on resistant varieties to overcome these challenges. In this study, towards improving yield, an effective protocol for attaining high frequency somatic embryogenesis (SE) in green gram has been proposed. Type of explants and age of source seedlings for obtaining explants were found to influence the formation of embryogenic calli. Various combinations and concentrations of 2,4-dichlorophenoxyacetic acid and indole-3-acetic acid with kinetin were optimized for developing embryogenic calli. Embryogenic calli when exposed to osmotic stress created by D-mannitol and sorbitol and dehydration stress imposed by polyethylene glycol were found to produce somatic embryos. Calli incubated for 6 h in specified hormone free nutrient medium supplemented with 4 % polyethylene glycol was optimal for induction of high frequency SE. Subsequent to stress incubation, the cultures formed only early stage somatic embryos. Supplementation of proline was found essential for the maturation of somatic embryos. Cotyledonary stage somatic embryos were converted into plantlets and subsequently established in garden soil. Semi-quantitative Reverse Transcription-PCR based transcript level analysis of SERK1 gene expression was carried out during different developmental stages of somatic embryogenesis. Expression of SERK1 was specifically associated with the embryogenic calli and calli enriched with globular stage somatic embryos.

2 illus, 4 tables, 39 ref

025499   NAMASIVAYAM S K R, ROBIN A T G (Biotechnology Dep, Sathyabama Univ, Chennai, Tamil Nadu, Email: biologiask@gmail.com) : Preparation of nano albumin-flutamide (Nab-flu) conjugate and evaluation of its in vitro drug control release, anticancer activity and genotoxicity. Indian J Exp Biol 2018, 56(3), 171- 9.

Cancer is one of the most common noncommunicable diseases of mankind which causes considerable deaths worldwide. Though there have been consistant efforts on prevention and control, cancer still ranks second in global mortality, causing one out of every six deaths. Nanotechnology has radically changed the way cancer is diagnosed, imaged and treated. Researchers have designed novel nanodevices capable of detecting cancer at its earliest stages, pinpointing its location within the body and delivering drugs specifically to malignant cells. Albumin, with active tumor targeting capacity, is a versatile drug carrier in anticancer drug delivery system. In this study, preparation of BSA nanoparticles has been optimized with various parameters such as pH, ethanol to BSA ratio and crosslinking time in order to improve drug delivery. The optimal pH was found to be 8.0, the ethanol to albumin ratio was found to be 4:1 and cross linking time of 8 h which gives the higher yield of BSA nanoparticles. Nanodrug conjugate was prepared using the optimized conditions and the nanospheres formed were characterized using SEM which showed a particle size of nanosphere in the range of 160-230 nm. Fourier transform infrared spectroscopy (FTIR) analysis showed the possible functional groups of nano drug conjugate. The drug loading efficiency and entrapment efficiency were found to be 81 and 83 %, respectively. The in vitro drug release profile was studied by continuous dialysis method. Cumulative release reached almost 81 % after 24 h and showed an almost released ability of the nanoparticle formulation. Sustained and controlled release profile of flutamide facilitates application of nanoparticles for delivery of anticancer drugs. The Nab-Flu, Nab was found to have very low toxicity on Vero cell line and a higher toxicity in hep2 cell lines. Further, genotoxicity study revealed the Nab and Nab-Flu showed a zero genotoxicity.

6 illus, 2 tables, 30 ref

024562   AL-JBORY Z , ANDERSON K M, HARRIS M O, MITTAPALLI O, WHITWORTH R J, CHEN M-S (Entomology Dep, Kansas State Univ, Kansas- 66506, United States, Email: mchen@ksu.edu) : Transcriptomic analyses of secreted proteins from the salivary glands of wheat midge larvae. J Insect Sci 2018, 18(1), 17.

Both the wheat midge (Sitodiplosis mosellana) (Géhin) (Diptera: Cecidomyiidae) and the Hessian fly (Mayetiola destructor) (Say) (Diptera: Cecidomyiidae) belong to a group of insects called gall midges (Diptera: Cecidomyiidae), and both are destructive pests of wheat. From Hessian fly larvae, a large number of genes have been identified to encode secreted salivary gland proteins (SSGPs), which are presumably critical for the insect to feed on and manipulate host plants. For comparison, we conducted an analysis on transcripts encoding SSGPs from the first instar larvae of the wheat midge. In total, 3,500 cDNA clones were sequenced, from which 1,301 high-quality sequences were obtained. Approximately 25 % of the cDNAs with high-quality sequences encoded SSGPs. The SSGPs were grouped into 97 groups based on sequence homology. Among the SSGP-encoding transcripts, 206 encoded unique proteins with no sequence similarity to any known protein and 29 encoded proteins similar to known proteins including proteases, serpines, thioesterases, ankyrins, and ferritins. Most (~80 %) SSGP-encoding genes appear under strong selection for mutations that generate amino acid changes within the coding region. Identification and characterization of SSGPs in wheat midge larvae provide a foundation for future work to reveal molecular mechanisms behind wheat midge–wheat interactions and the role of these putative effector proteins in insect virulence. Availability of the SSGP transcripts will also facilitate comparative analyses of insect effectors from related species.

1 illus, 1 table, 25 ref

024561   WEN R, WANG B, WANG B, MA L (Northeast Forestry Univ, Harbin- 150 040, P R China, Email: maling63@163.com) : Characterization and expression profiles of juvenile hormone epoxide hydrolase from Lymantria dispar (Lepidoptera: Lymantridae) and RNA interference by ingestion. J Insect Sci 2018, 18(1), 13.

Juvenile hormone epoxide hydrolase (JHEH) is an important enzyme in the degradation pathways of juvenile hormone (JH) in insects. It converts JH to JH diol and hydrolyses JH acid to JH acid diol. JHEH titers regulate the entire process of insect development. In this study, full length ldjheh cDNA (2101 bp) was cloned from the Asian gypsy moth Lymantria dispar (L.; Lepidoptera: Lymantridae), and provisionally designated ldjheh1. LdJHEH1 was characterized by predicted molecular weight of 52.64 kDa, theoretical isoelectric points of 6.87 and contains a transmembrane domain at the N-terminus. The transcriptional profiles of ldjheh1 were detected by qRT-PCR. The ldjheh1 was found to be expressed throughout all developmental stages with maximum expression levels occurring in fourth instar larvae. The ldjheh1 mRNA was detected in the heads, thoraces, and abdomens of gypsy moth larvae on day 2 of the third instar. The ldjheh1 was also detected in bodies of third instar larvae stage, with the highest peaks occurring at 24 h after ecdysis. The ldjheh1 gene was successfully knocked down by oral delivery dsRNA in the third instar larvae of L. dispar. The dsRNA targeting ldjheh1 was produced in vitro. Ingesting dsRNA for ldjheh1 only slightly delayed larval development.

7 illus, 1 table, 46 ref

024560   XU K, NIU Q, ZHAO H, DU Y, GUO L, JIANG Y (Animal Genetics and Breeding & Reproduction Dep, Shanxi Agricultural Univ, Shanxi- 030 801, China, Email: jiangys-001@163.com) : Sequencing and expression characterization of antifreeze protein maxi-like in Apis cerana cerana. J Insect Sci 2018, 18(1), 11.

Antifreeze proteins (AFPs) are biological cryoprotectants with unique properties that play a crucial role in regulating the molecular mechanisms governing cold resistance in insects. To identify and characterize Apis cerana cerana AFP (AcerAFP), we cloned the full-length cDNA of AcerAFP and examined its expression patterns in A. cerana cerana. A nucleotide alignment analysis showed that the entire coding region of AcerAFP is 1095 bp and encodes a polypeptide of 365 amino acids. The amino acid sequence of this protein exhibits 63–96 % homology with AFP homologs from other hymenopterans. α-helices form the main secondary and tertiary structures of AcerAFP, which is similar to the molecular structure of fish AFP type-I. The expression profiles of AcerAFP revealed that expression was tissue, sex, and developmentally specific. In response to cold stress, the mRNA and protein expression of AcerAFP were both induced by low temperatures, and were also related to the concentrations of several cryoprotectants, including glucose, glycerin, glutamic acid, cysteine, histidine, alanine, and methionine. In addition, we found that the knockdown of AcerAFP by RNA interference remarkably increased the total freezing temperature of hemolymph in A. cerana cerana, where levels of AcerAFP mRNA were correlated with the expression of most antifreeze-related proteins. Taken together, these results suggest that AcerAFP plays an essential role as a biological cryoprotectant in honeybees, and is in turn regulated by small cryoprotectants and antifreeze-related proteins.

7 illus, 1 table, 48 ref

024559   SHAO E, LIN L, LIU S, ZHANG J, CHEN X, SHA L, HUANG Z, HUANG B, GUAN X (Fujian Agriculture and Forestry Univ, Fujian, PR China, Email: guanxfafu@126.com) : Analysis of homologs of Cry-toxin receptor-related proteins in the midgut of a non-Bt target, Nilaparvata lugens (St?l) (Hemiptera: Delphacidae). J Insect Sci 2018, 18(1), 10.

The brown planthopper (BPH) Nilaparvata lugens is one of the most destructive insect pests in the rice fields of Asia. Like other hemipteran insects, BPH is not susceptible to Cry toxins of Bacillus thuringiensis (Bt) or transgenic rice carrying Bt cry genes. Lack of Cry receptors in the midgut is one of the main reasons that BPH is not susceptible to the Cry toxins. The main Cry-binding proteins (CBPs) of the susceptible insects are cadherin, aminopeptidase N (APN), and alkaline phosphatase (ALP). In this study, we analyzed and validated de novo assembled transcripts from transcriptome sequencing data of BPH to identify and characterize homologs of cadherin, APN, and ALP. We then compared the cadherin-, APN-, and ALP-like proteins of BPH to previously reported CBPs to identify their homologs in BPH. The sequence analysis revealed that at least one cadherin, one APN, and two ALPs of BPH contained homologous functional domains identified from the Cry-binding cadherin, APN, and ALP, respectively. Quantitative real-time polymerase chain reaction used to verify the expression level of each putative Cry receptor homolog in the BPH midgut indicated that the CBPs homologous APN and ALP were expressed at high or medium-high levels while the cadherin was expressed at a low level. These results suggest that homologs of CBPs exist in the midgut of BPH. However, differences in key motifs of CBPs, which are functional in interacting with Cry toxins, may be responsible for insusceptibility of BPH to Cry toxins.

6 illus, 2 tables, 57 ref

024558   JIN T, GAO Y, HE K, GE F (Chinese Academy of Sciences, Beijing, China, Email: gef@ioz.ac.cn) : Expression profiles of the trehalose-6-phosphate synthase gene associated with thermal stress in Ostrinia furnacalis (Lepidoptera: Crambidae). J Insect Sci 2018, 18(1), 7.

Trehalose is the major blood sugar in insects. Physiological significance of this compound has been extensively reported. Trehalose-6-phosphate synthase (TPS) is an important enzyme in the trehalose biosynthesis pathway. Full-length cDNAs of TPS (of TPS) and its alternative splicing isoform (Oftps_isoformI) were cloned from the Asian corn borer (ACB), Ostrinia furnacalis (Guenée; Lepidoptera: Crambidae) larvae. The Oftps and Oftps_isoformI transcripts were 2913 and 1689 bp long, contained 2529 and 1293 bp open reading frames encoding proteins of 842 and 430 amino acids with a molecular mass of 94.4 and 48.6 kDa, respectively. Transcriptional profiling and response to thermal stress of Oftps gene were determined by quantitative real-time PCR showing that the Oftps was predominantly expressed in the larval fat body, significantly enhanced during molting and transformation; and thermal stress also induced Oftps expression. Gene structure analysis is indicating that one TPS domain and one trehalose-6-phosphate phosphatase (TPP) domain were located at the N- and C-termini of OfTPS, respectively, while only the TPS domain was detected in OfTPS_isoformI. Three-dimensional modeling and heterologous expression were developed to predict the putative functions of Of TPS and OfTPS_isoformI. We infer that the expression of Oftps gene is thermally induced and might be crucial for larvae survival.

6 illus, 1 table, 41 ref

024557   RAMAN K V, AGGARWAL D, RAO S R, SREEVASTHA R, SINGH A K, ABDIN M Z, MOHAPTRA T, PATTANAYAK D (ICAR-National Research Centre on Plant Biotechnology (NRCPB), New Delhi- 110 012, Email: debasispattanayak@yahoo.co.in) : Rapid and efficient agrobacterium mediated transformation of early scutellum derived calli of indica rice. Indian J Expl Biol 2018, 56(1), 20- 8.

Rice is a staple food for humans and its demand in 2035 has been put at 852 million tons. Knowledge on genes and genome architecture helps in better understanding of growth and development mechanisms for crop improvement. Transgenic crops may offer a solution by means of higher yield and resistance to biotic and abiotic stresses. In this context, modification of Agrobacterium mediated transformation protocol for indica rice cultivar is imperative to increase transformation efficiency and reduce duration of transgenic development. Here, we developed an efficient Agrobacterium mediated transformation protocol using early scutellum derived calli of the indica rice cultivar Pusa Sugandh 2. Competency of 3, 4, 5 and 6 day old primary calli was compared with 21-day old secondary calli for Agrobacterium mediated transformation using a modified pCAMBIA 1304 harbouring GFP-GUS fusion gene driven by maize ubiquitin 1 promoter. The highest competency with stable transformation efficiency of 51 % was observed for 5-6 day old primary calli. Molecular analysis confirmed stable integration of the transgene. Transgenic lines of Pusa Sugandh 2 were developed within a short period of two months using 5-6 day old primary calli.

6 illus, 3 tables, 45 ref

024556   PACHAURI P, MORE S, ARANGANATHAN V, SULLIA S B, DESHMUKH S (Biotechnology Dep, Jain Univ, Bangalore – 560 011, Email: sudhadeshmukh@yahoo.com) : Kinetic study and characterization of cellulase enzyme from isolated Aspergillus niger subsp. awamori for cellulosic biofuels. J Sci Ind Res 2018, 77(1), 55-60.

In the present study, a cellulase producing Aspergillus species was isolated from wood chips and was identified as Aspergillus niger subsp. awamori (also called A. awamori) using 18s rRNA sequencing (Genbank Accession number KP164968). The enzyme was purified to 42.5 fold by gel filtration chromatography and the molecular mass of the enzyme was determined as 66±1 kDa using SDS-PAGE. The purified enzyme showed an optimum temperature of 50°C and pH 5. Metal ions such as Ca²⁺, Mg²⁺, Zn²⁺, Pb²⁺, Fe²⁺ and Fe³⁺ had positive influence on the enzyme activity. The K_M and V_max for the enzyme was found to be 0.011 g and 0.1098 U/ml respectively. The addition of this cellulase to lignocellulosic biomass showed good degradation of cellulose and therefore this fungus could be a future source for production of bioethanol from Lignocellulosic Biomass (LB).

3 illus, 3 tables, 25 ref

024555   KANDIYIL S, MALEK R A, AZIZ R, ENSHASY H A E (Technology Malaysia Univ, Johor, Email: henshasy@ibd.utm.my) : Development of an industrially feasible medium for enhanced production of extremely thermophilic recombinant endo-1,4-β-xylanase by Escherichia coli. J Sci Ind Res 2018, 77(1), 41-9.

Escherichia coli BL21 (DE3) with a plasmid vector pET-22b (+) carrying xylanase coding gene isolated from an extremely thermophilic bacterium, Thermotoga neapolitana was used in current study for the production of recombinant endo-1,4-β-xylanase (EC 3.2.1.8) which is also called xylanase in short. The study was focused on development of an industrially feasible production media for enhanced production of xylanase. Plackett–Burman based model was initially applied to identify the significant media components effecting the xylanase production, followed by a linear full factorial experimental design (General Linear Model), to estimate the optimum concentration ranges of those significant components and the interactions between them. Finally, Box-Behnken based design was applied to fine tune the concentration rages of significant media components as well as the inducer. Using response maximizing tool, up to 878.72 IU mL^-1 of xylnase activity was predicted in media composition (in g L^-1): Maltose, 22.8; (NH₄)H₂ PO₄,10.5; K₂HPO₄, 16.5; KH₂PO₄, 3.6; MgSO₄.7H₂O, 2; and trace element solution (TES), 3.8 mL L^-1 when induced with 8 g.L^-1 of lactose at mid log phase of cell growth. During the predicted model validation, the intracellular xylanase production was found enhanced up to of 800 IU mL^-1 which was around 3 folds increase in productivity when compared to IPTG induced production in unoptimized medium. As a part of scaling up the process, the optimized media composition and cultivation conditions were then tested in pilot scale bioreactor and up to 5600 IU mL^-1 of xylnase activity was achieved at the end of batch cultivation.

3 illus, 3 tables, 24 ref

024554   KUMAR S S, JITHIN V, JIJEESH V, GAYATHRI V, SHIBURAJ S, HARIDAS M, SABU A (Biotechnology and Microbiology Dep, Kannur Univ, Kerala - 670 661, Email: drsabu@gmail.com) : Production and purification of alkaline protease from Exiguobacterium indicum TBG-PICH-001 isolated from soil samples of Pichavaram estuary (Tamil Nadu). Indian J Geo-Mar Sci 2018, 47(03), 580-6.

A proteolytic bacterial strain, TBG-PICH-001, was isolated from soil of Pichavaram estuary, Tamil Nadu, and was identified as Exiguobacterium indicum based on 16S rDNA sequence similarity. Parameters such as incubation time, temperature, initial pH, rate of agitation and inoculum required for peak production of the protease by the newly isolated bacterium under submerged fermentation were optimized using one factor at a time method. Maximum protease production was observed after 48 hrs of incubation at 35 ˚C with constant stirring at 100 rpm, at an initial pH of 10. Reduced incubation time for enzyme production shows the industrial viability of the strain. Purification of enzyme was achieved by ammonium sulphate fractionation followed by ion-exchange chromatography with DEAE-Cellulose, to give total yield of 27 %. The molecular weight determined by SDS- PAGE was approximately 60 kDa.

9 illus, 1 table, 31 ref

024553   KUMAR S S, GOVARDHAN T L, HEMALATHA K P J (Microbiology and Research Centre PG Dep, Dr. Lankapalli Bullayya Post-graduate Coll, Visakhapatnam-530 013, Email: ksuribabu_sda@yahoo.com) : Thermal stability of partially purified α-amylase produced by Brevibacillus borstelensis R1 from the coastal waters of Bay of Bengal, Visakhapatnam. Indian J Geo-Mar Sci 2018, 47(02), 437-42.

Highest α-amylase activity of Brevibacillus borstelensis R1 was observed at 37 ⁰C (6133 ± 58 U/ml). The α-amylase retained its activity after 10 hrs exposure to 4 ⁰C with negligible variations in its activity. Alpha-amylase have several applications in starch processing, desizing of textiles, paper sizing, detergent additive, bread improvement, ethanol production, sewage treatment, effluent treatment and other fermentation processes. Highest α-amylase activity was observed at 37 ˚C (6133 ± 58 U/ml), while the lowest at 70 ˚C (2360 ± 40 U/ml). Only 38.47 % activity remained at 70 ˚C. Optimum temperature of partially purified enzyme was similar to the crude enzyme. Partially purified enzyme was tested for its stability (1-10 hours) at temperatures ranging from 4 ˚C to 70 ˚C. At 20 ˚C it retained 34.2 % activity, at 25˚C 44.1 %, at 37 ˚C 34.7 %, at 50 ˚C 56.2 %, at 60 ˚C 38.2 % and at 70 ˚C it retained 30.8 % activity upon exposure for about 10 hrs.

8 illus, 38 ref

024552   CHEIRSILP B, SUKSAWANG S, YEESANG J, BOONSAWANG P (Industrial Biotechnology Dep, Prince of Songkla Univ, Songkhla- 901 12, Email: benjamas.che@psu.ac.th) : Co-production of functional exopolysaccharides and lactic acid by Lactobacillus kefiranofaciens originated from fermented milk, Kefir. J Food Sci Technol 2018, 55(1), 331-40.

Kefiran is a functional exopolysaccharide produced by Lactobacillus kefiranofaciens originated from kefir, traditional fermented milk in the Caucasian mountains, Russia. Kefiran is attractive as thickeners, stabilizers, emulsifiers, gelling agents and also has antimicrobial and antitumor activity. However, the production costs of kefiran are still high mainly due to high cost of carbon and nitrogen sources. This study aimed to produce kefiran and its co-product, lactic acid, from low-cost industrial byproducts. Among the sources tested, whey lactose (at 2 % sugar concentration) and spent yeast cells hydrolysate (at 6 g-nitrogen/L) gave the highest kefiran of 480 ± 21 mg/L along with lactic acid of 20.1 ± 0.2 g/L. The combination of these two sources and initial pH were optimized through Response Surface Methodology. With the optimized medium, L. kefiranofaciens produced more kefiran and lactic acid up to 635 ± 7 mg/L and 32.9 ± 0.7 g/L, respectively. When the pH was controlled to alleviate the inhibition from acidic pH, L. kefiranofaciens could consume all sugars and produced kefiran and lactic acid up to 1693 ± 29 mg/L and 87.49 ± 0.23 g/L, respectively. Moreover, the fed-batch fermentation with intermittent adding of whey lactose improved kefiran and lactic acid productions up to 2514 ± 93 mg/L and 135 ± 1.75 g/L, respectively. These results indicate the promising approach to economically produce kefiran and lactic acid from lowcost nutrient sources.

6 illus, 21 ref

024551   TAGAD C K, SABHARWAL S G (Chemistry Dep, Savitribai Phule Pune Univ, Pune- 411007, Email: ssab@chem.unipune.ac.in) : Purification and characterization of acid phosphatase from Macrotyloma uiflorum seeds. J Food Sci Technol 2018, 55(1), 313-20.

Acid phosphatases play a crucial role in food processing industries to reduce phosphate content of food. Here in acid phosphatase from the seeds of Macrotyloma uniflorum has been purified to homogeneity using UNOsphere- S cation exchange chromatography followed by gel filtration with 81.85 fold purification. Molecular weight of purified enzyme was 55,000 (± 1040) Daltons under physiological conditions. It was a heterodimer of subunits having molecular weights 27,093 and 28,241 Daltons as determined by MALDI-TOF analysis. The optimum pH and temperature for the purified enzyme was 5.0 and 50 ⁰C respectively. The enzyme was stable in the pH range 3.5–5.5 and showed temperature stability up to 60 ⁰C. Substrate specificity of enzyme was checked with different substrates namely, p-nitrophenyl phosphate (p-NPP), ATP, ADP, glucose 6-phosphate, glucose-1-phosphate, fructose 6-phosphate, phenyl phosphate, a-naphthyl-phosphate, pyridoxyl phosphate and b-glycerophosphate. Enzyme showed high substrate specificity towards p-NPP, phenyl phosphate, ATP and a-naphthyl phosphate. Km and Vmax of enzyme were found to be 0.934 mM and 1.333 mM/min respectively with respect to p-NPP as a substrate. Chemical modification studies showed that tryptophan was present at the active site of the enzyme.

4 illus, 24 ref

024550   VERMA N, KAUR N, KRISHNAN P (Biotechnology Dep, Punjabi Univ, Patiala - 142 001, Email: verma.neelam2@gmail.com) : Application of molecular beacon based biosensor against rs699 SNP in hypertensive and non-hypertensive Punjabi population. Int J Pharmac 2018, 5(1), 37-50.

Molecular beacons have shown their topnotch potential in a variety of basic research, biomedical detection and clinical diagnosis. Their excellent selectivity, sensitivity, detection without separation have made them widely accepted tool for nucleic acid analysis. MBs present high-throughput screening of SNPs. The use of SNPs in the detection of genetic disorders is facilitated by the recent discovery of more than 4 million SNPs in the human genome. The objective of this study was to explore the association of rs699 SNP with essential hypertension in Punjabi population. MB based biosensor developed by Verma et al., in 2016 was chosen for analysis. These hypertensive subjects tested positive against hypertensive biosensor (against allele C) were associated with essential hypertension. C4072T or rs699 polymorphism was genotyped in 50 hypertensive and 50 normotensive subjects. The p-value for the C allele in hypertensive patients was 0.71, which concludes susceptibility to hypertension in this population when the CC genotype is present. The implementation was cross-validated by applying the same samples to Normotensive Biosensor (against T allele). The presence of fluorescence with hypertensive biosensor confirms the presence of a mutation in hypertensive patients or patients that are clinically naive but can develop hypertension in the forthcoming due to hypertensive allele presence. Hence it can help in the future pharmacogenetic based treatment of patients.

5 illus, 11 tables, 48 ref

024549   PATIL M Y, SAWANT G B, JADHAV S M (Agric Botany Dep, Dr. Balasaheb Sawant Konkan Krishi Vidyapeeth, Dapoli, Email: 007mansinghpatil@gmail.com) : Combining ability and gene action studies in chilli (Capsicum annuum L.). Environ Ecol 2018, 36(1), 52-6.

A study was undertaken to estimate combining ability in nine diverse genotypes of chilli by following diallel cross analysis (excluding reciprocals). Thirty six hybrids along with nine genotypes were evaluated during 2011–2012. All the characters studied showed significant variability among the genotypes. For the combining ability analysis, the variances due to g.c.a were significant for all the characters except number of primary branches per plant showed non-significant s.c.a variance. Additive gene action was observed for all the characters studied except for days to first and days to 50 % flowering where non additive gene action were found. Among the parents Pant C-3, BC-28, CO-4 and RHRC- 16-5 were found good general combiner for fruit yield and yield contributing characters. Hybrids BC-28 × Pant C-3, RHRC-16-5 × BC-28, RHRC-16-5 × CO-4 and RHRC-16-5 × Pant C-3, RHRC-16-5 × Konkan Kirti and BC-28 × Konkan Kirti were found to be having good s.c.a effects for fruit yield and contributing characters and useful in further breeding program.

3 tables, 7 ref