006808   LUTHFI M, INDRAWATI R, OKI A S, RIFA’I M (Oral Biology Dep, Airlangga Univ, Surabaya, Indonesia, Email: m.luthfi@fkg.unair.ac.id) : Complement receptor 1 (CR1)/ CD35⁺ expression analysis of salivary neutrophils on Streptococcus mutans phagocytosis. Contemp Clin Dent 2019, 10(4), 627-30.

Severe early childhood caries (S-ECC) is a form of dental caries which is very destructive in early childhood since involving several teeth, include the maxillary anterior teeth. Streptococcus mutans (S. mutans) play an etiological integral role of ECC so that S. mutans are considered as the predictor of dental caries. The neutrophil is a key component of the rst line of defense against microbial invasion. The essential function of neutrophil is to kill pathogenic microbes through a phagocytosis process which is mediated by Complement Receptor 1 (CR1)/ (CD35⁺ ). To analyze the phagocytosis process of the salivary neutrophil which is mediated by innate immunity component, i.e., Complement Receptor 1/CR1 (CD35) on S-ECC. his study was an observational analysis with cross-sectional approach using t-test analysis. This study employed the isolation steps of neutrophils saliva of cariesfree children and the S-ECC and then conducted phagocytosis of salivary neutrophils test on S. mutans mediated by CD35 using ow cytometry. Results: Phagocytosis of salivary neutrophils on S. mutans mediated by CD35 on caries-free (2.35 ± 0.56) is higher than that on the S-ECC (1.54 ± 0.35). It is concluded that there is a decrease of phagocytic on S. mutans mediated by Complement Receptor 1/CR1 (CD35+) on S-ECC.

2 illus, 1 table, 17 ref

006797   EWIDA A Y I, MOHAMED W S E-D (Microbiology Dep, National Water Research Center, Cairo, Egypt, Email: aymanyi@yahoo.com) : Isolation and characterization of biosurfactant producing bacteria from oil-contaminated water. Biosci Biotech Res Asia 2019, 16(4), 833-41.

Biosurfactants are chemical compounds produced by some microorganisms to initiate oil biodegradation. They have been applied generously in many industries. The present work aimed to isolate and identify a new bacterial strain, of water habitat, capable of producing biosurfactant. So, water samples were collected from three different water environments including river Nile at Alkanater city, Qalyubia governorate; was representing clear raw water. River Nile at ship settlement station, Imbaba city, Giza governorate; was representing oilcontaminated water, where there were some oil spills from ship fixation. Rahawy drain, Giza governorate; wasrepresentinghighly pollutedwastewater. The bacterial community of each water environment was isolated and inventoried, then screened for biosurfactant production by blood hemolysis, oil spreading technique, drop collapse assay, foaming activity and emulsification activity. Bacterial strains isolated from the oil-contaminated environment showed high potential for biosurfactant production, and the best biosurfactant producing isolate was identified by 16S rRNA technique as Pseudomonas protegens, and the produced biosurfactant was belong to rhamnolipid group.

4 illus, 2 tables, 35 ref

006793   BEYAHMEDOV I A (Guba Regional Agrarian-Science and Innovation Center, Azerbaijan, Europe) : The effects of grafting-variety combinations to every year productivity indicators and effective fruit fallling on pear plant. Biosci Biotech Res Asia 2019, 16(4), 827-31.

In the article is assigned information about useful fruit falling and productivity every year in the combinations different grafting-varieties of the pear. The research was carried out in the Guba and Gusar regions of the north-eastern of country. The researches were carried out; in the Dakhli village of Guba region, on the grafting wild forest pear, 6x4 m food area, with Khure which was planted in 2004; The sorts as beauty of Talgar, white Williams, beauty of forest in the New life village of Qusar region on the graftin BA-29, 4x1.25m food area, planted in 2008-2009; The varieties as conference, white Williams, red Williams, Forelle and Quyot which were cultivated in the trellis and has been installed drip irrigation system The research was carried out by generally accepted methods for gardening. At this time, every two years productivity index was calculated by a Sinq formula. According to the index varieties and years are considered: 20 % - up indicator - highly stable harvestable; 21 ... 40 % - to compared stable product each year; 41 ... 60 % - Average degree of every two years harvestable; 61 ... 80 % strong harvestable every two years; 81 ... 100% more powerful harvestable every two years.

1 illus, 1 table, 16 ref

006813   RAHMAWATI E, ABINAWANTO, HELIANTI I (Biology Dep, Indonesia Univ, Jawa Barat, Indonesia, Email: is.helianti@bppt.go.id) : Subcloning and expression of a protease gene from Bacillus halodurans CM1 in Bacillus subtilis DB104. Biosci Biotech Res Asia 2019, 16(4), 817-26.

Proteases are potential enzymes that utilized in various industrial fields, and the demand of these enzymes is increasing. Bacillus halodurans CM1 is Indonesia indigenous bacterium which is detected to be able to produce alkalotermophilic protease enzyme. In this study, we subcloned the protease gene consist of Open Reading Frame of protease gene and its promoter from Bacillus halodurans CM1 in Bacillus subtilis DB104 via conjugation, and analyzed the expression of the recombinant protease. The protease gene is 1 417 bp length including the open reading frame and the promoter, and obtained by PCR and cloned into pGEM T easy. After confirmed by sequencing, the gene was subcloned into vector pBBRE194, then the recombinant plasmid was transformed into E. coli S17^-1. This E.coli was then conjugated to Bacillus subtilis DB104. The target recombinant B. subtilis DB104 has been obtained confirmed by plasmid verification and erythromycin resistance. The recombinant protease produced showed the highest enzyme activity at 50  ^oC and pH 9 (with pH range 5-9) which with protease activity 13.66 U/mL.

5 illus, 31 ref

006824   SUHAIL M, PARVEEN A, HUSAIN A, REHAN M (The Glocal Univ, Uttar Pradesh- 247 121, Email: mrehan786@gmail.com) : Exploring inhibitory mechanisms of green tea catechins as inhibitors of a cancer therapeutic target, nuclear factor-κB (NF-κB). Biosci Biotech Res Asia 2019, 16(4), 715-23.

Nuclear factor kappa B (NF-êB), a transcription factor is a well-established cancer therapeutic target. NF-êB’s linkage with cancer is known through the constitutive activation of NF-êB in several cancer types. The most important role of NF-êB as a transcription factor is its ability to promote cell survival through the induction of transcription of target pro-survival genes and thus inhibition of programmed cell death (PCD) by resulting proteins in both malignant and normal cells. Current findings have unveiled that green tea catechins exert anticancer effect by inhibiting the activity of various receptors including NF-êB. The current study is designed to gain the structural insights for inhibitory mechanism of catechin derivatives against NF-êB. The major green tea catechins include (-)-epicatechin (EC), (-)-epigallocatechin (EGC), (-)-epicatechin gallate (ECG), and (-)-epigallocatechin gallate (EGCG) and are included in the current study. The study explored the binding pose, interacting residues, molecular interactions, and predicted binding energy and dissociation constant for the catechin derivatives. Our results showed that the catechin derivatives bound well in the DNA binding site with adequate binding strength scores. The study suggested that the four catechin derivatives may act as potential inhibitors of NF-êB and thus, may inhibit the progression of various cancer types.

5 illus, 6 tables, 39 ref

006829   WANIG H, YU Z, GUI Z, LIU W, YE S, CHU W (Pharmacy Dep, Wenzhou Medical Univ, Lishui- 323 000) : Multidrug resistance, extracellular enzymatic activity and biofilm formation of Staphylococcus aureus isolates from various animal foods in East China. Biomed Res 2019, 30(6), 1-6.

Staphylococcus aureus is the most common pathogenic bacteria that causes staphylococcal food poisoning, a form of gastroenteritis with rapid onset of symptoms. The development of multiple drug resistance to this organism is posing serious threat to human health; these bacteria also have plenty of virulence factors which contribute to infection. Ampicillin, penicillin, and amoxicillin were the least effective. Norfloxacin, cephalexin, ciprofloxacin, rifampicin, clindamycin and vancomycin were the most effective antibiotics by in vitro sensitivity testing. S. aureus isolates from foodstuff were producers of a variety of extracellular hydrolytic drug resistance.

3 tables, 32 ref

006798   GUPTA R, KUMAR T, MITTAL A (Biotechnology Dep, Kurukshetra Univ, Kurukshetra- 136 119, Email: amitakuk@gmail.com) : Bioremediation of cadmium contaminated effluent by Sporosarcina luteola: A bacterium isolated from soil near Wazirpur Industrial Area, New Delhi, India. Asian J Chem 2019, 31(11), 2642-6.

Heavy metals pollution is emerging as a threat to ecological systems causing various problems to mankind, plants and animals. Aim of the present study was to isolate and identify cadmium tolerant bacteria from the soil of Wazirpur industrial area of New Delhi (India). The study involved physico-chemical characterization of the polluted soil which was found to contain high concentration of iron, manganese and cadmium at 352, 15.3 g/kg soil and 3.16 ppm, respectively. One bacterial strain was identified as Sporosarcina luteola on the basis of morphological, biochemical and phylogeny analysis. Strain Sporosarcina luteola was highly resistant to Cd up to 5mM (mM= millimolar) when cultured in solidified nutrient agar plates and 7.2 mM in nutrient broth. Sporosarcina luteola has also showed substantial growth in presence of Co, Pb, Fe and Mn upto 2.0, 2.0, 3.5 and 4.0 mM, respectively in liquid medium. Optimum growth of identified bacteria was shown at 37 ºC, 7.0 pH and it tolerated up to 3 % sodium chloride (w/v). This is reported for the first time that Sporosarcina luteola (metal-tolerant bacteria) has potential of removal of cadmium from industrially contaminated soil.

2 illus, 2 tables, 30 ref

006832   ZAIED B K, SIDDIQUE M N I, ZULARISAM A W, AHMAD M F, SALIH Y M (Malaysia Pahang Univ, Pekan- 26600, Email: m.nurul@umt.edu.my) : Biomethane production from anaerobic codigestion of palm oil mill effluent with cattle manure: A review. Asian J Chem 2019, 31(11), 2413-24.

Biogas is conventionally utilized in a gas engine to convert the chemical energy into electricity and into thermal energy for heating purposes and is also pumped into the natural gas grid line after impurities are removed. Biogas production from palm oil mill effluent is one of the best options for economic expansion and this would reduce environmental impacts in developing countries like Malaysia. This study aims to perform an analysis of the economic and environmental prospects of biogas production from the anaerobic codigestion method from palm oil mill effluent mixed with cow manure. Anaerobic codigestion technology is considered a practical process by which to overcome the complications of substance properties and system optimization in particular substance digestion operations. Comparisons among various treatment technologies of palm oil mill effluent and their applications have been studied thoroughly. The factors that affect biogas production, along with strategies for their improvement, were studied in present report. The economic feasibility of biogas plant where palm oil mill effluent and cattle manure are main substrates and anaerobic codigestion is the method, has been successfully predicted. Results revealed this might be more feasible if this technology is used on a large scale. Construction of the proposed biomethanization plant is economically feasible because it is projected that about a four-year return-on-investment will be achieved. In conclusion, the present work demonstrates a comprehensive feasibility framework by which to integrate the different features needed to enhance biomethane generation.

1 illus, 6 tables, 103 ref

006816   RANI A, AZMI W (Biotechnology Dep, Himachal Pradesh Univ, Shimla- 171 005, Himachal Pradesh, Email: wamikazmi@rediffmail.com) : An overview on biosynthesis and applications of extracellular pyocyanin pigment and its role in Pseudomonas aeruginosa pathogenesis. Ann Phytomed 2019, 8(2), 28-42.

Microbial pigments are chemical moieties capable of absorbing light in the visible range. The demand for natural colorants is increasing in every day as the present trend throughout the world is shifting towards the use of ecofriendly and biodegradable commodities. Among the natural sources, the pigment producing microorganisms hold a promising potential to meet present day challenges. Pigments are compounds that come in a wide variety and extensively used in industries. Industrial production of natural pigments by microbial fermentation has several advantages over extraction of pigments of plant origin, such as easy availability of inexpensive raw materials for their production, independence from crop seasons and their seasonal variations, etc. The Pseudomonas aeruginosa has ability to produce a number of redox-active phenazine compounds including pyocyanin. The pyocyanin pigment possesses antimicrobial, anticancerous, antioxidant, anti-inflammatory and immunosupperessive properties. This review covers the challenges and new insights into pyocyanin from P. aeruginosa with emphasis on the role of pyocyanin in P. aeruginosa infection with special attention to applications of pyocyanin pigment.

6 illus, 4 tables, 187 ref

006814   RAJATHY P L L, VARSHNEY K C, NAIR M G, LAKKAWAR A W, KUMAR B R (Veterinary Surgery and Radiology Dep, Rajiv Gandhi Institute of Veterinary Education and Research, Kurumbapet- 605 009, Pondicherry, Email: leenavet@gmail.com) : A study on immunohistochemical expression of progesterone receptor and cell proliferation by AgNOR in Canine Mammary Tumors (CMT). Agric Sci Dig 2019, 39(4), 335-40.

Mammary gland tumors are the most commonly diagnosed tumors in domestic dogs. Although immunohistochemical methods provide valuable information such as the location and semi-quantitative data of the interested antigens in particular tumors, conventional methods like histopathological diagnosis remain useful and necessary for identification and classification of tumors. In the present study, we combined histopathology with immunohistochemical staining for progesterone receptor (PR) and special staining argyrophilic nucleolar organizer regions (AgNOR) for cell proliferation in canine mammary gland tumors. Twenty-nine dogs with primary mammary gland neoplasms (11 benign and 18 malignant) were included in this study. All 29 tumors were stained with AgNOR but only 15 cases expressed positive PR signals in the nuclei of neoplastic alveolar and ductal epithelial cells. A weak positive correlation (r = 0.2) was observed between PR and AgNOR index of benign tumors and a very weak negative correlation (r = -0.1833) between PR and AgNOR index of malignant tumors suggesting that higher cellular proliferation is associated with low expression of progesterone receptors. 

28 ref

006802   KAVIPRIYA C, YUVARAJA A, SENTHIL K, MENAKA C (Plant Breeding and Genetics Dep, Agricultural Coll and Research Institute, Madurai- 625 104, Email: yugenetics@yahoo.com) : Genetic transformation methods for crop improvement- A brief review. Agric Rev 2019, 40(4), 281-8.

Decades of documented successful pieces of evidence in agritech have shown us a clear picture of the importance of biotechnology in crop improvement. The production in agriculture should have steady growth and to achieve these objective conventional methods should go on parallel with biotechnological approaches. Genetic engineering which has revolutionized the path of crop improvement involves the identification and transfer of novel genes into the existing elite cultivars. Different methods of transferring the gene into plant cells have been developed and continuous efforts have been made to increase its efficiency. Both direct and indirect method of gene transfer has its own merits and demerits. Efforts have been made continuously to eliminate drawbacks and to develop an easy, elite and eco-friendly method to transfer genes. The transformation method which is a base of genetic engineering is vital and Agrobacterium-mediated gene transfer and gene gun have shown to be doing well in recent years. As a whole the methodology involved, merits and demerits of different methods have been briefly discussed.

7 illus, 2 tables, 53 ref

005528   PANDA B B, MEHER A S, HAZRA R K (Biotechnology Dep, Ravenshaw Univ, Cuttack, Odisha, Email: rupenkh@yahoo.co.in) : Comparison between different methods of DNA isolation from dried blood spots for determination of malaria to determine specificity and cost effectiveness. J Parasit Dis 2019, 43(3), 337–42.

DNA extraction from filter paper by using different methods was compiled through a thorough review of many research articles published in various journals. When performing malaria epidemiological surveys in remote area, it is difficult to collect blood samples and transport it. In field particularly in remote area where facilities for storing and processing of samples does not exist, there surveillance and diagnosis of malaria is very difficult. In this review we are focused upon four simple methods of DNA isolation from the field collected blood and mosquito abdomen blood meal spotted on Whatman No. 1 or No. 3 filter paper. The main DNA isolation methods are Chelex100, Tris-EDTA (TE) buffer; Methanol based DNA extraction and Phosphate buffer saline (PBS) using Lysis buffer and Phenol–Chloroform method. Efforts have been taken to identify the methods which are cost-effective and take less time to extract DNA from dried blood spots (DBS) and whole mosquitoes. The purpose of this paper is to update the knowledge and find a method to extract DNA from DBS which will be specific, rapid, cost-effective, less time consuming and feasible for epidemiological survey in remote area.

1 table, 41 ref

005553   VU T, RAZANSKY D, YAO J (Biomedical Engineering Dep, Duke Univ, NC, United States of America, Email: daniel.razansky@uzh.ch) : Listening to tissues with new light: Recent technological advances in photoacoustic imaging. J Opt 2019, 21(10), 103001.

Photoacoustic tomography (PAT), or optoacoustic tomography, has achieved remarkable progress in the past decade, benefiting from the joint developments in optics, acoustics, chemistry, computing and mathematics. Unlike pure optical or ultrasound imaging, PAT can provide unique optical absorption contrast as well as widely scalable spatial resolution, penetration depth and imaging speed. Moreover, PAT has inherent sensitivity to tissue’s functional, molecular, and metabolic state. With these merits, PAT has been applied in a wide range of life science disciplines, and has enabled biomedical research unattainable by other imaging methods. This review article aims at introducing state-of-the-art PAT technologies and their representative applications. The focus is on recent technological breakthroughs in structural, functional, molecular PAT, including super-resolution imaging, real-time small animal whole-body imaging, and high-sensitivity functional/molecular imaging. We also discuss the remaining challenges in PAT and envisioned opportunities.

5 illus, 1 table, 15 ref

005531   PATEL H A, GAJERA R R (Food Processing Technology and Bio-Energy Dep, AAU, Anand, Gujarat) : Effect of steam blanching on quality of bottle gourd shreds. J Med Plants Stud 2019, 7(5)-A, 1-3.

Bottle gourd (Lagenaria siceraria) is an important medicinal herb and vegetable crop of tropical and subtropical regions of the world belonging to the family Cucurbitaceae. Blanching inactivates the prevailing enzymes in the food which are responsible for the product quality loss. Hot water and steam are the most commonly used heating methods for blanching in industry. This study was undertaken to investigate the effect of steam blanching on bottle gourd shreds after inactivation of peroxidase enzymes. During blanching, the enzyme inactivation time was found 6.00, 7.50 and 8.30 min, in 3, 5 and 7 mm bottle gourd shreds, respectively. The quality parameters of bottle gourd shreds at various thickness and time with their interactions were studied. The total soluble solids was increased with blanching time increases and decreased with shreds thickness increases, however the combined effect was found non-significant. The ascorbic acid content was decreased significantly as shreds thickness and blanching time increases and their interaction were found highly significant (P<0.05). Heat sensitive ascorbic acid reduction was found more in 7 mm shreds after steam blanching. The juice yield was found increased significantly as shred thickness increases and decreased as blanching time increases indicating significant influence of independent parameter.

2 tables, 11 ref

005544   SARUTA F, YAMADA N, YAMAMOTO K (Bioscience and Biotechnology Dep, Kyushu Univ, Fukuoka 819-0395, Japan, Email: yamamok@agr.kyushu-u.ac.jp) : Functional analysis of an epsilon-class glutathione S-Transferase from Nilaparvata lugens (Hemiptera: Delphacidae). J Insect Sci 2019, 19(5), 96.

Glutathione conjugation is a crucial step in xenobiotic detoxification. In the current study, we have functionally characterized an epsilon-class glutathione S-transferase (GST) from a brown planthopper Nilaparvata lugens (nlGSTE). The amino acid sequence of nlGSTE revealed approximately 36–44 % identity with epsilon-class GSTs of other species. The recombinant nlGSTE was prepared in soluble form by bacterial expression and was purified to homogeneity. Mutation experiments revealed that the putative substrate-binding sites, including Phe107, Arg112, Phe118, and Phe119, were important for glutathione transferase activity. Furthermore, inhibition study displayed that nlGSTE activity was affected by insecticides, proposing that, in brown planthopper, nlGSTE could recognize insecticides as substrates.

4 illus, 3 tables, 33 ref

005520   KUMBICAK Z (Molecular Biology and Genetics Dep, Nevsehir Haci Bektas Veli Univ, Nevsehi- 50300, Turkey, Email: zkumbicak@nevsehir.edu.tr) : First cytogenetic analysis of Eratigena agrestris (Araneae: Agelenidae) from Turkey. J Insect Sci 2019, 19(5), 86.

In this study, chromosomal characteristics of Eratigena agrestris (Agelenidae) were investigated for the first time. Karyotype features including diploid chromosome number, sex chromosome system, chromosome morphology, and meiotic behavior were obtained from specimens collected in two localities of Mediterranean region. A spreading method including dissection, hypotonization, fixation, and staining was used to prepare the chromosome slides. In a total, 10 adult males were used due to having high numbers of dividing cells. Cytogenetical results showed that the diploid chromosome number and sex chromosome system was 2n♂ = 42 (X₁ X₂ 0). The sex chromosomes were identified tentatively. All chromosomes were telocentric. Relative chromosome lengths of autosomal pairs ranged between 5.65 and 3.32 %, and relative chromosome lengths of X₁ and X₂ were 5.33 and 4.19 %, respectively. In the first meiotic division stages, bivalents usually had one chiasma, but some had two chiasmata. At the end of the meiosis, two kinds of nuclei, with or without sex chromosomes, have occurred. These results contribute to a better characterization of the Agelenidae cytogenetic.

2 illus, 1 table, 13 ref

005550   TRIPATHI A, MELO J S (Nuclear Agriculture and Biotechnology Div, Bhabha Atomic Research Centre, Mumbai- 400 085, Email: anujtri@barc.gov.in) : Cryostructurization of polymeric systems for developing macroporous cryogel as a foundational framework in bioengineering applications. J Chem Sci 2019, 131(9), 92.

Among various fabrication techniques to produce a porous scaffold, thermally-induced phase separation at controlled cryogenic condition leads to the formation of a porous polymeric cryostructure alias cryogel or cryomatrix. Cryostructurization is one of the simple and versatile methods of synthesizing a highly porous and interconnected architecture. The process of cryostructurization is present in comparable fabrication approach of fabricating advanced porous biomaterials with precise control over multiple compositions of precursor units, spatial distributions for accomplishing effective recapitulation of mechanical properties and architectural accuracy at micron-scale with bioactive functionality. The cryogenically-structured polymeric scaffolds are of noteworthy fundamental and applied interest in multi-disciplinary areas of science and showed promising matrices in various biotechnological and biomedical areas. The large interconnected pores in cryogels open up a range of applications like the three-dimensional substrate for cell growth, bioreactor for continuous production of scarce molecules, bio-processing and protein purification, an adsorbent for environmental remediation, and biosensor fabrication. Over the past two decades, significant attention to these materials with rigorous expansion in their new designs, and remarkable growth in the number of the publications and patents on cryostructured porous materials has been noticed. This article is covering the progress in cryostructurization technology in the last few decades with principles underlying the mechanism of cryogelation, process optimization, and the recent trend in cryogel for biomedical and bioengineering applications.

7 illus, 1 table, 57 ref

005504   HUSSAIN A, VERMA C K (Bioinformatics Dep, Maulana Azad National Institute of Technology, Bhopal, Madhya Pradesh, Email: ahussain591@gmail.com) : Ligand- and structure-based pharmacophore modeling, docking study reveals 2-[[4-[6-(isopropylamino) pyrimidin-4-yl]-1H-pyrrolo[2,3-b] pyridin-6-yl] amino] ethanol as a potential anticancer agent of CDK9/cyclin T1 kinase. J Can Res Ther 2019, 15(5), 1131-40.

CDK9/Cyclin T1 kinase is a protein kinase, indirectly involved in the cell cycle progression in the form of transcription elongation, CDK9 specific inhibitors may be a potential alternative treatment not only for cancer but also other life‑threatening diseases. Ligand‑based and structure‑based pharmacophore model was developed for discovering of the new anticancer agents. These models used as three‑dimensional query for virtual screening against the chemical structure databases such as Maybridge HitFinder, MDPI, and ZINC. Subsequently, the potential hit compound was filtered by the ADMET and docking score. After applying all filtration, 11 hits were found as potential hits based on good docking scores as well as good ADMET properties. Compound 2‑[[4‑[6‑(isopropylamino) pyrimidin‑4‑yl]‑1H‑pyrrolo[2,3‑b] pyridin‑6‑yl] amino] ethanol was found to be most potent among all the potential hits. These hits could be used as an anticancer agent in near future. So many advances in the treatment of death leading diseases have been made over the past few decades, However, looking for the development in this research ligand‑based and structure‑based pharmacophore modeling was done, hit1 2‑[[4‑[6‑(isopropylamino) pyrimidin‑4‑yl]‑1H‑pyrrolo[2,3 b] pyridin‑6 yl] amino] ethanol was found to be more potent and selective. It is understandable that these hits could be as selective and potent anticancer agents of cyclin‑dependent kinase complex.

9 illus, 5 tables, 44 ref

005556   YADAV B S, SINGH S, SRIVASTAVA S, MANI A (Biotechnology Dep, Motilal Nehru National Institute of Technology, Allahabad- 211 004, Email: amani@mnnit.ac.in) : Analysis of chickpea gene co-expression networks and pathways during heavy metal stress. J Biosci 2019, 44(4), 99.

Crop productivity and yield are adversely affected by abiotic and biotic stresses. Therefore, finding out the genes responsible for stress tolerance is a significant stride towards crop improvement. A gene co-expression network is a powerful tool to detect the most connected genes during heavy metal (HM) stress in plants. The most connected genes may be responsible for HM tolerance by altering the different metabolic pathways during the biotic and abiotic stress. In the same line we have performed the GSE86807 microarray analysis of chickpea during exposure to chromium, cadmium and arsenic and analyzed the data. Common differentially expressed genes (DEGs) during exposure to chromium, cadmium and arsenic were identified and a co-expression network study was carried out. Hub and bottleneck genes were explored on the basis of degree and betweenness centrality, respectively. A gene set enrichment analysis study revealed that genes like haloacid dehydrogenase, cinnamoyl CoA reductase, F-box protein, GDSL esterase lipase, cellulose synthase, b-glucosidase 13 and isoflavone hydroxylase are significantly enriched and regulate the different pathways like riboflavin metabolism, phenyl propanoid biosynthesis, amino acid biosynthesis, isoflavonoid biosynthesis and indole alkaloid biosynthesis.

3 illus, 8 tables, 57 ref

005526   NAWKARKAR P, SINGH A K, ABDIN M Z, KUMAR S (International Centre for Genetic Engineering and Biotechnology, New Delhi-110 067, Email: skrhode@icgeb.res.in) : Life cycle assessment of Chlorella species producing biodiesel and remediating wastewater. J Biosci 2019, 44(4), 89.

Constantly rising energy demands, finite fossil fuel reserves and deteriorating environmental conditions have invoked worldwide interest to explore the sustainable sources of renewable biofuels. Locally adapted photosynthetic oleaginous microalgae with rapid growth on variable temperatures could be an ideal way for bioremediating the wastewater (WW) while producing the feedstock for biodiesel. To test this notion, an unknown strain was isolated from a sewage fed lake (Neela-Hauz). It was discerned as Chlorella sorokiniana-I using the 16S rDNA and 18S rDNA barcodes. The culture conditions such as pH, illumination, different temperature ranges and growth medium were cohesively optimized prior to the assessment of C. sorokiniana-I’s efficacy to remediate the WW and biodiesel production. The strain has thrived well up to 40 ⁰C when continuously grown for 15 days. The highest lipid accumulation and biomass productivity were recorded in 100 % WW. Fatty acid methyl ester (FAME) content was observed to be more than twice in WW (47 %), compared to control synthetic media, TAP (20 %) and BG11 (10 %), which indicate the importance of this new isolate for producing economically viable biodiesel. Moreover, it is highly efficient in removing the total nitrogen (77 %), total phosphorous (81 %), iron (67 %) and calcium (42 %) from the WW. The quality of WW was considerably improved by reducing the overall chemical oxygen demand (48 %), biological oxygen demand (47 %) and alkalinity (15 %). Thus, C. sorokiniana-I could be an ideal alga for the tropical countries in the remediation of WW while producing feedstock for biodiesel in a cost-effective manner.

5 illus, 5 tables, 68 ref

005484   BALAKRISHNAN B, DAVID E (Biotechnology Dep, Thiruvalluvar Univ, Vellore- 632 115, Email: ernestdavid2002@yahoo.com) : Biopolymers augment viral vectors based gene delivery. J Biosci 2019, 44(4), 84.

The success of viral vectors mediated gene therapy is still hampered by immunogenicity and insufficient transgene expression. Alternatively, non-viral vectors mediated gene delivery has the advantage of low immunogenicity despite showing low transgene expression. By carefully considering the advantages of each approach, hybrid vectors are currently being developed by modifying the viral vectors using non-viral biopolymers. This review provides an overview of the hybrid vectors currently being developed.

4 illus, 55 ref

005552   VERMA H, RANI A, GAHLOT M, GOEL A, SHARMA A K (Clothing and Textiles Dep, Govind Ballabh Pant Univ of Agriculture and Technology, Pantnagar, Udham Singh Nagar, Email: verma.himani017@gmail.com) : Sustainable dyeing of wool fabric with Talaromyces purpurogenus. J Appl Nat Sci 2019, 11(4), 796-801.

Natural dyes had been used by human being since ancient times for colouring of various materials including textiles. These were replaced completely by synthetic dyes, continuous use of which created the problem of pollution and environmental degradation. With the development of green technology and increased awareness of sustainability, the use of non-allergic, non-toxic and eco-friendly natural dyes on textiles has become a matter of significant importance. Among the natural sources of colourants, microorganisms such as bacteria and fungi have gained interest in the field of textile coloration.  In the present study,  the  pigment  solution  yielded  from  growth  of Talaromyces purpurogenus sp.  on organic waste was used for dyeing of wool fabric. Under the dyeing conditions i.e. 1:30 M:L ratio, pH 5, 80º C temperature and 45 minutes dyeing time, the percent absorption could reach 57 %, 32 %, 36 % and 49% respectively. The wool fabric samples dyed with the fungal dye exhibited very good to excellent washing and rubbing fastness properties. The colour pigment obtained from fungal thus offered an opportunity to reduce impact on plant resources for dye source exploration. So it can be concluded from the study that a natural  fungal  dye  could  help  to  sustain  the  environment  and  to  minimize  the  over exploitation of natural resources.

8 tables, 35 ref

005492   CHOLASSERY S, KRISHNA V, SETHURAJ S V, REHINA S S, RANGANATHAN V, DILEEP L C, SABU K R, RAJESH B R, CHANDRAN R P (Biotechnology and Research Dep, K. V. M. Coll of Science and Technology, Kerala- 688 527, Email: drpratapchandran@yahoo.co.in) : Analysis of physicochemical and sensory parameters of wine produced from Carica papaya. J Appl Biol Biotechnol 2019, 7(5), 74-8.

Wine is one of the most ancient beverages and it consists of a complex mixture of alcohol, sugars, aldehydes, tannins, pectins, vitamins, minerals, and organic acids formed by the action of Saccharomyces cerevisiae on fruit juices. Papaya (Carica papaya) is a common plant originally from South America and grown mainly in Asian and African countries. In the present study, C. papaya fruit pulp was used for the production of wine under controlled fermentation using S. cerevisiae. The physicochemical parameters such as pH, yeast cell count, specific gravity, carotenoid content, and alcohol content were observed during the study. The analysis was done once every 5 days. Fourier Transform Infrared Spectroscopy analyses of wine and unfermented fruit juice samples also were carried out. During fermentation, the pH was changed from 5.1 to 4.4. The initial value of yeast cell count during the fermentation process was 8.5×10⁶ cells/ml, later it attained the maximum value of 9.53×10⁶ cells per ml on the 5th day. It then gradually decreased to a final value of 2.92×10⁶ cells/ml on the 20th day. The specific gravity was initially 1.1573 and it decreased to 1.1483. Carotenoid content was 0.034 mg/ml on the first day and it increased to 0.279 mg/ml. Alcohol content of papaya was found to be increased during the course of fermentation. It was estimated as 3.01% on the 5th day and it increased to 10.11% on the 20th day. The overall point for the sensory analysis was 4.8.

4 illus, 1 table, 18 ref

005532   PATEL R, DASGUPTA D, VERNEKAR M (D. Y. Patil Univ, Navi Mumbai, Maharashtra, Email: madhavi.vernekar@dypatil.edu) : High-density culture of vero cells in novel perfusion bioreactor system. J Appl Biol Biotechnol 2019, 7(5), 50-5.

Improvements in upstream bioprocessing single-use systems have allowed implementation of high-density cultures in standard workflows. This study shows the capability of the novel designed single-use bioreactors to accommodates very high surface area per unit volume and enable the cell culture scientists to achieve and maintain a stable, high-density perfusion culture of Vero cells, without compromising mixing characteristics and nutritional homogeneity. Conventionally, adherent cell lines have been cultivated in non-controlled, labor-intensive, and space-demanding 2D surface technologies, i.e., roller bottles (RB) and multi-layer flasks. Fixed-bed and Packed-bed bioreactors are popular alternatives to culture high-density adherent cell lines for vaccines and other biologics production. However, the limitations that restricts the use of fixed-beds are nonhomogeneous cell seeding, huge mass transfer gradients, inability of maintaining nutritional homogeneity, limitation of bed size expansion and difficulties during scale-up of etc. In this study, we have shown the design and development of novel disc-based bioreactors in single-use format for high-density Vero cell culture. During the process run of 12 days, we have observed 4.97 population doublings and final cell density achieved was 0.438 million cells/sq.cm from the seeding density of 0.014 million cells/sq.cm. Vero cell growth observed in novel bioreactor was significantly higher than the traditional culture devices, such as RB and multilayer flasks.

7 illus, 2 tables, 14 ref

005522   LOONG S-K, LEE H-Y, KHOO J-J, LIM F-S, AHMAD-NASRAH S-N, AZMAN A-S, SUNTHARALINGAM C, PANCHADCHARAM C, ABUBAKAR S (Malaya Univ, Kuala Lumpur- 50603, Email: sazaly@um.edu.my) : Microbiological analysis of raw milk unveiled the presence of a dairy contaminant, Corynebacterium lipophiloflavum. J Appl Biol Biotechnol 2019, 7(5), 41-4.

Dairy farming occupied a distinct position in agriculture since milk can be harvested every day, providing a regular source of income to the farmers. Development of the Malaysian dairy farming industry was marred by poor farm hygiene practices, leading to the proliferation of dairy-spoilage bacteria, affecting milk quality. In this study, we report the isolation and characterization of a rare Corynebacterium species from raw milk after the implementation of improved farm hygiene practices. All milking equipment, farm worker’s hands and the cow’s udders and teats were washed with detergent and wiped dry with clean towels before milk sample collection. Collected foremilk samples from mastitis-free cows were inoculated onto Petrifilm™ and cultured colonies were plated onto nutrient agar. Biochemical and molecular tests were performed for the identification of peculiar bacterial isolates. A unique yellow-pigmented bacteria isolate was recovered from the milk of a healthy cow after the adoption of improved farm hygiene practices. Phenotypic and genotypic characterization confirmed the milk isolate as Corynebacterium lipophiloflavum. This is the first description of C. lipophiloflavum in cow’s milk and could possibly imply the influence of bovine flora in dairy contamination. The findings highlight the increasing spectrum of Corynebacterium species with potential adverse impact to the dairy industry. It is recommended to screen for C. lipophiloflavum in all milk processing facility to ensure that milk is safe for consumption and its products prepared to the highest quality and safety standards.

2 illus, 1 table, 19 ref

005517   KOUR D, RANA K L, YADAV N , YADAV A N (Biotechnology Dep, Eternal Univ, Baru Sahib, Sirmour, Email: ajar@eternaluniversity.edu.in) : Bioprospecting of phosphorus solubilizing bacteria from Renuka lake ecosystems, lesser himalayas. J Appl Biol Biotechnol 2019, 7(5), 1-6.

In the present study, diversity and plant growth promoting traits of bacteria isolated from Renuka Lake, Himachal Pradesh was investigated. A total of three samples, including sediment (pH-7.3), sub-surface water (pH-8.1), and surface water (pH-8.2), were collected. A total of 65 bacterial isolates were obtained on different growth media. Among 65 isolates 18 isolates were found to be solubilizers of phosphorus. The maximum amount of phosphorus was solubilized by EU-RL 54 (7,976 ± 0.01 µg/l) followed by EU-RL 53 (6,322 ± 0.01 µg/l). Phosphorus solubilizers also possessed other plant growth promoting traits, such as the production of ammonia, hydrogen cyanide, zinc solubilization, production of hydrolytic enzymes. The isolates were identified belonging to different genera Acinetobacter, Bacillus, Enterobacter, Klebsiella, Proteus, Pseudomonas, and Staphylococcus. This is the first report for these plant growth promoting bacteria to solubilize a considerable amount of phosphorus isolated from Ecosystems, Lesser Himalayas.

3 illus, 1 table, 30 ref

005490   BORAL R, RATHORE R S, MISHRA A K, KUMAR A (Veterinary Public Health Dep, Indian Veterinary Research Institute, Bareilly- 243 122, Email: boralrupa@gmail.com) : Sensitivity and specificity of multiplex PCR assay for detection of Yersinia enterocolitica in pigs and foods of porcine origin. Vet Pract 2019, 20(2), 153-8.

Yersinia enterocolitica, an important food and water-borne gastrointestinal agent is regarded an emerging pathogen world wide. Animals that recover frequently become asymptomatic carriers of the disease. Most human illness is caused by Y. enterocolitica which causes a variety of symptoms depending on the age of the person infected often in young children. In the present study, a polymerase chain reaction (PCR) based assay was used for rapid detection of Y. enterocolitica targeting 16S rRNA, ail and yst genes in pig faeces, tongue, tonsils and ground pork samples. The primers were found to be highly specific for Y. enterocolitica and did not yielded any amplification with other Gram positive and Gram negative bacteria. The minimum detection level was found up to 103 cells/ml for all the genes targeted in present assay. Prevalence rates were 5.83 % in pig faeces; 6.67 % in pig tongues; 18.3 % in pig tonsils; 6.67 % in pork, respectively. Overall prevalence rate was reported 8.34 % with highest prevalence found in pig tonsils and lowest in pig faeces. PCR technique was found to be more efficient with 97.5 % sensitivity than the cultural methods with only 90 % sensitivity. Comparison studies revealed that the PCR assay was more rapid, specific, sensitive and efficient over conventional cultural and biochemical detection methods. To conclude, we could say that the PCR based detection is not only a sensitive but also a rapid and reliable method for detection of Y. enterocolitica in different sample matrices as clearly demonstrated by the results in present study.

3 illus, 4 tables, 42 ref

005495   DINESH R M, PATEL A K, VIBHA J B, SHEKHAWAT S, SHEKHAWAT N S (Botany Dep, Jai Narain Vyas Univ, Rajasthan- 342 001) : Cloning of mature pomegranate (Punica granatum) cv. Jalore seedless via in vitro shoot production and ex vitro rooting. Vegetos 2019, 32(2), 181–9.

A novel approach of in vitro shoot amplification and ex vitro rooting for cloning of mature plants of Punica granatum cv. Jalore seedless/soft-seeded has been defined. Surface-sterilized nodal shoots were cultured for axillary meristem activation, bud breaking and shoot amplification. Multiple shoots differentiated by bud breaking from 82.8% of the explants on Murashige and Skoog (Physiol Plant 15:473–497, 1962) MS medium with 13.32 µM 6-benzylaminopurine (BAP). These were amplified by subculturing of fresh shoots and by repeated transfer of mother explants on MS medium + BAP or Kinetin (Kin)/Furfurylaminopurine (FAP) in combination with Indole acetic acid (IAA) or α-naphthalene acetic acid (NAA). MS medium with BAP 2.22 µM + IAA 0.57 µM was found to be most suitable for shoot (14.2 ± 1.03 shoots per culture vessel) multiplication. On half-strength MS medium with 9.84 µM of Indole butyric acid (IBA) and 16.65 µM of activated charcoal (AC), 72.9 % of the micro-shoots rooted. Alternatively, base(s) of the micro-shoots treated with 1476 µM of IBA for 300 s. and transplanted on autoclaved soilrite (moistened with one-fourth strength of MS macro-salts) in glass bottles (420 ml; 70 mm diameter × 130 mm height). More than 85 % of the IBA-treated shoots rooted ex vitro in a greenhouse. Ex vitro rooting of cloned shoots is a new approach for propagation of pomegranate. The process described is different and superior to all the described tissue culture methods for cloning of pomegranate. This is faster, cost-effective and saves resources enabling acclimatization/hardening with ease while minimizing microbial contamination, thus ensuring quick field transfer of hardened plantlets. This can be applied for mass and clonal propagation of selected genotypes and also for long-term conservation of germplasm of P. granatum.

70 ref

005521   LAHA S, KUMAR D, SENGUPTA D N, GANGOPADHYAY G (Botany Dep, Bethune Coll, Kolkata- 700 006) : In silico characterization of SAMdC from Pokkali rice and its overexpression in transgenic tobacco. Vegetos 2019, 32(2), 158-66.

Polyamines help to mitigate salt stress in plants. SAMdC or S-adenosyl methionine decarboxylase is a crucial enzyme in the polyamine biosynthetic pathway. Overexpressing the samdc gene of Pokkali (a salt-tolerant Indica rice landrace) in the model plant tobacco through Agrobacterium-mediated transformation resulted in tobacco plants capable of withstanding high salinity (200 mM) stress. The transgenic tobacco plants maintained a steady level of the higher polyamines and resisted ionic imbalance under salt stress. Since appropriate binding of the substrate with the enzyme is essential for an enzymatic reaction, we performed molecular docking experiment of SAMdC enzyme of Pokkali rice and tobacco to get an idea about its commonality in the two plants in the backdrop of Japonica rice and Arabidopsis. In silico characterization of SAMdC revealed that the enzyme used the same substrate in Pokkali rice and tobacco, from where the gene is introgressed and where it is overexpressed.

20 ref

005508   JANGRA M R, VERMA C, CHARAYA G, MANGAL M, SAINI K, JANGRA S, NEHRA K S (Biotechnology Dep, Government Coll, Hisar-125 004) : Development and characterization of Kinnow wine with and without pulp using activated yeast. Vegetos 2019, 32(1), 110-5.

We report here the attempts made to prepare and characterize wine from Kinnow. Kinnow is gaining more commercial significance and attractiveness due to its success under varied agro climatic conditions. Its unique characterstics like high fruit yield, fruits quality and wider adaptability make it a very good choice for post harvest processing. Must prepared from Kinnow show very good fermentation when inoculated with sacchromyces cerevisae. Kinnow wines were prepared and evaluated for various physio–chemical characteristics like antioxidant activity, color, ethanol, pH, free and total sulfur dioxide, titrable acidity and enological properties. So, the present investigation was carried to find out the optimal conditions for the efficient conversion of orange juice with or without pulp into wine using Saccharomyces cerevisiae. The optimum process conditions for this fermentation process were 21°, 22° and 24° Brix (Total Soluble Solids), 25 °C temperature, 10% inoculum, and pH in between 3 and 5. Corresponding to these optimum conditions, the predicted value of ethanol production was found to be 11–12%, which was experimentally verified. Four samples A, B, 3 and 4 have shown more antioxidant activity as compared to sample C, D, 1 and 2. On the basis of sensory and nonsensory evaulaution, it was found that wines were found to dry, still, orange in color, acidic and acceptable for drinking purposes.

24 ref

005489   BHATT P P, THAKER V S (Biosciences Dep, Saurashtra Univ, Rajkot- 360 005) : Extremely diverse structural organization in the complete mitochondrial genome of seedless Phoenix dactylifera L. Vegetos 2019, 32(1), 92-7.

Mitochondrial genome of seedless (mutant) variety of Phoenix dactylifera is analyzed to understand the probable reason for seedless condition. The size of the genome is 585,493 bp long which is 129,508 bp shorter compared to the normal genome and demonstrate significantly higher rate of nucleotide loss. The loss is evident in coding and non-coding regions. Missing of all rRNA genes and other three genes, i.e., rps14, rps13, rps19 is observed. Total 2726 SNPs are recorded, showed 80.9 % identity with the normal genome. The ratio of nonsynonymous (dN) synonymous (dS) substitution (dN/dS) is more than one for nad7, nad1, ccMb, ccmC, cob, matR, matB suggest adaptive selection while less than one for cox2 suggest strong selective pressure on this gene and reduction in size may be start of its transfer to nucleus. Such variations may play an important role in seedless condition is discussed.

38 ref

005530   PARWANI L, BATNAGAR M, BHATNAGAR A, SHARMA V, SHARMA V (Bioscience and Biotechnology Dep, Banasthali Vidyapith, Newai- 304 022) : Gum acacia-PVA hydrogel blends for wound healing. Vegetos 2019, 32(1), 78-91.

Hydroactive wound dressings ensure a physiologically moist wound milieu which enhances healing and patient comfort. Polymers like polyurethane, salts of alginic acid, collagen, cellulose and other gelable polysaccharides are commonly used for preparation of hydrogels but have their own limitations. In the present study acacia gum, a natural polysaccharide having known antioxidant and wound healing properties has been blended with polyvinyl alcohol and prepared hydrogel matrix further explored for its wound healing potential. Structural characterization of blended films by FTIR and XRD method showed that polysaccharide gum associated with PVA molecules via acetal bridges and they were amorphous in nature. The gel exhibited free swell capacity of 64 g 0.100 cm⁻², which is comparable to the commercially available films recommended for heavily exuding wounds. The GA/PVA blends showed 34 % moisture retention (Rh) ability after 24 h and their fluid absorbing (26 %) and fluid donation (16 %) ability make them suitable for moist and fibrinous wounds. They also exhibited blood compatibility, oxygen permeability, bacterial impermeability, antioxidant activity, iron chelation ability and biodegradability. In vivo healing potential has been evaluated on Swiss albino mice where hydrogel coated wounds showed faster and scar-less wound regeneration. In vitro Povidone-Iodine loading and release studies showed that iodine release followed Fickian diffusion process. Prepared hydrogels are breathable and hydrophilic in nature and are able to maintain appropriate moisture level at wound surface, suitable for acceleration of wound healing process. Based upon the studied properties the designed hydrogels are recommended for dry, necrotic and low exuding wounds.

69 ref

005557   YADAV N, GAUTAM B, YADAV P K (Computational Biology and Bioinformatics Dep, Sam Higginbottom Univ of Agriculture, Prayagraj- 211 007) : Computational analysis of microarray-based gene expression profiling and unveiling the functional traits in the developmental phases of Glycine max seed. Vegetos 2019, 32(1), 64-77.

The Glycine max seed is an ultimate source of protein worldwide and plays the crucial role in crop development. This made us curious to insight the developmental stages of G. max seed at the genomic level with computational based microarray data analysis. The data analysis explored 10453 and 6085 significant DEGs from 0 to 4 mm vs. 12 to 16 mm and 4–8 mm vs. 8–12 mm stage, respectively and based on log2 FC, 407 & 2342 DEGs were regulating high expression and 340 & 1414 DEGs were low in expression at the 0–4 mm vs.12–16 mm and 4–8 mm vs. 8–12 mm stage, respectively. Gene symbols for DEGs has recognized and identified that, 7 DEGs from 0–4 mm vs. 12–16 mm stage were coding for miRNAs. Gene classification searched 2 & 4 highly enriched groups of genes from up-down regulated DEGs of 0–4 mm vs. 12–16 mm stage, respectively and 19 & 6 groups of genes from up-down regulated DEGs of 4–8 mm vs. 8–12 mm stage that were showing functional similarities. Analysis for functional characterization has found that 38 % & 36 % DEGs were annotating for BP, 42 % & 67 % DEGs were for CC and 53 % & 46 % DEGs were for MF in up-down regulated DEGs of 0–4 mm vs. 12–16 mm stage, respectively. For up-down regulated DEGs of 4–8 mm vs. 8–12 mm stage has drawn and 8 % &10 % DEGs were indicating BP, 4 % & 3 % for CC and 15 % & 23 % DEGs were annotating for MF, respectively. Top ten highly regulated genes with the ID 11998843, 11855572, 12189716, etc. from stage 0–4 mm vs. 12–16 mm were annotating 10 BP, 6 MF and 6 CC. While, up-regulated DEGs with the ID Gma.10969.1.S1_at, GmaAffx.92715.1.S1_s_at, etc. were annotating 19 MF, 7 BP and one term from CC for stage 4–8 mm vs. 8–12 mm. These analyses would be helpful in the further characterization of interesting candidate genes that involve in the seed development of G. max.

27 ref

005554   WAHLANG D R, SUCHIANG W, GOEL S, RAO S (Biotechnology and Bioinformatics Dep, North-Eastern Hill Univ, Shillong- 793 022) : Analysis of genetic variation using ISSR and the development of SCAR marker in synthetic autotetraploids of Vigna mungo (L.) Hepper. Vegetos 2019, 32(1), 48-57.

Autopolyploidy is one of the foremost mechanisms by which additional numbers of genome copies are acquired resulting in genetic and evolutionary novelty to the organisms. As such, it is considered to be a pathway for adaptation and has played a significant role in plant speciation. Autopolyploids are characterized by increased genome flexibility, allowing them to adapt and persist across heterogeneous landscapes in the long run. Genomic redundancy and polysomic inheritance are the hallmarks of doubled genome(s). In view of the significance of autopolyploidy in plant evolution, plant breeding and crop improvement, we sought to investigate the genetic variation/s associated with genome multiplication by exploiting artificial somatic autopolyploids in Vigna mungo. ISSR was employed to detect and identify any genomic DNA variation in colchitetraploids of V. mungo of three different generations, viz. C₀, C₁ and C₂, in comparison to their putative diploids plants. Our data suggests a random, ‘short-term’ rapid change of the genome that may have occurred immediately after polyploidization. The population of autopolyploids demonstrated some degree of genetic heterozygosity in comparison to their diploid counterparts. Further, a polymorphic ISSR DNA loci was identified among the colchitetraploids which was translated into SCAR marker for identification of colchitetraploids versus the putative diploids of V. mungo.

36 ref

005543   RUSTAGI A, SHEKHAR S, KUMAR D, LAWRENCE K, BHAT V, SARIN N B (Jawaharlal Nehru Univ, New Delhi- 110 067) : High speed regeneration via somatic embryogenesis in elite Indian banana cv. Somrani monthan (ABB). Vegetos 2019, 32(1), 39-47.

An efficient and rapid regeneration system is indispensable for germplasm conservation and valuable trait incorporation through genetic engineering. The present study was aimed to investigate regeneration potential of Musa sp. cv. Somrani monthan (ABB) via somatic embryogenesis. Meristematic shoot tips were used as explant and cultured on MS medium supplemented with 6-BAP (50 and 100 µM) and IAA (1 µM) for scalp (cauliflower like compact buds) induction. The highest percentage (~ 60 %) of homogenous differentiation of shoot tip explants into scalp was observed in MS medium supplemented with 100 µM of 6-BAP and 1 µM of IAA after 6 months of culture. The scalps were sub-cultured on MS medium supplemented with different compositions of 2, 4-D (1, 2, 3, 4, 5 and 10 µM) and zeatin (1 and 2 µM) for embryogenic calli induction. The highest frequency (97 %) of embryogenic calli induction was recorded in MS medium supplemented with 5 µM 2, 4-D and 1 µM zeatin after 6 weeks of culture. The histological and morphological studies suggest that, callus possessed high competence for embryogenesis and could be induced to form somatic embryos. Embryogenic cell suspension (ECS) was successfully established by culturing these embryonic calli in liquid medium. About 70 % of the mature somatic embryos germinated and converted to plantlets with a regeneration capacity of 8.5 × 10³. After the gradual acclimatization and hardening, the plants were transferred to nursery with 82% of survival rate. Here we report a successful regeneration system for an elite Indian Musa cv. Somrani monthan via somatic embryogenesis. The embryogenic callus and ECS are ideal explants for mass clonal propagation, germplasm conservation and genetic transformation for future research.

42 ref

005503   HRIDEEK T K, GEETHU P D, Jijeesh C M, RAGHU A V, MURALEEKRISHNAN K (Kerala Forest Research Institute, Kerala- 680 653, Email: drhrideek@gmail.com) : Standardization of adventitious root induction in stem cuttings of Cynometra travancorica Bedd willd., an endangered tree species of Western Ghats. Vegetos 2019, 32(1), 11-8.

Cynometra travancorica Bedd. belonging to family Fabaceae is an endangered tree species endemic to Western Ghats with restricted distribution. There is an urgent need for conservation of this endangered species and the vegetative propagation techniques can play major role in planting stock production. Present study focuses on the standardisation of vegetative propagation protocol of C. travancorica using stem cuttings with different concentrations of three plant growth regulating substances in different season. The anatomical, physiological and biochemical changes of the sprouted cuttings belonging to different growth regulator treatments were also conducted. Softwood, semi hardwood and hardwood cuttings were collected in three seasons (January–April, May–August and September–December) and treated with indole 3 butyric acid, indole 3 acetic acid and naphthalene acetic acid in different concentrations. Rooting response was measured after 45 days in the mist chamber. The adventitious rhizogenesis of C. travancorica stem cuttings was influenced by type of cutting, season of collection, type of growth regulator and its concentration. Semi hardwood cuttings collected during season III treated with 8000 mg l⁻¹ IBA recorded about 84% rooting. Anatomical studies on the sprouted cuttings treated with different growth regulator did not show any significant differences, whereas physiological and biochemical parameters of sprouted cuttings varied. Hence, we recommend this treatment combination for the large scale planting stock production.

5 illus, 4 tables, 23 ref

005539   RAMAKRISHNAIAH T N, ARUNKUMAR S (Biotechnology and Genetics Dep, Ramaiah Coll of Arts, Science and Commerce, Bangalore- 560 054, Email: ramakrishnaiahtn@gmail.com) : RAPD profiling of Oxyopidae spiders. Indian J Entomol 2019, 81(3), 541-5.

Random amplified polymorphic DNA (RAPD) Polymerase Chain Reaction (PCR) was used to characterize six spiders belonging to the family Oxyopidae. The aim was to achieve genetic marker to determine the genetic variations of these species. The species studied includes: Oxyopes lineatipes (C L Koch, 1847), O. javanus (Thorell, 1887), O. shweta (Tikader, 1970), O. sunandae (Tikader, 1970), Hamadruas sp. (Thorell, 1887) and Peucetia viridana (Stolickza, 1869) collected from Karnataka. RAPD-PCR was carried out by using three random primers which generated polymorphic fragments. Genetic polymorphism among these samples was recorded by Neighbour Joining (NJ) method. The results revealed that their genetic distance ranged from 0.6962- 0.8523. Cluster analysis based on this revealed that the tree is divided into two clades. The results concluded that the RAPD-PCR is an effective, fast and simple alternative method for identifying these spiders.

3 illus, 2 tables, 24 ref

005514   KHAN T A, PATHAK D, SHARMA P, ANSARI M, AGNIHOTRI R K (Botany Dep, Dr. B.R. Ambedkar Univ, Agra, Uttar Pradesh, Email: rk_agnihotri@rediffmail.com) : In vitro regeneration, callus induction and rhizogenesis in Ficus krishnae: A rare endangered plant. Indian J Biotechnol 2019, 18(4), 346-51.

Ficus krishnae which belongs to the family Moraceae is an endangered plant species with medicinal potential. Due to over-exploitative commercial use, it has become endangered hence demands immediate conservation. Application of in vitro strategies in conservation of plants is an efficient technique to produce true-to-type plantlets. In the present study, nodal segment explants were used as induction of shoots on Murashige & Skoog (MS) medium fortified with various concentrations of benzylaminopurine (BAP) in combination with 0.5 mg/l naphthalene acetic acid (NAA) to determine optimum levels of BAP and NAA for multiple shoot formation. Cytokinins, BAP and kinetin (Kn) were tested for shoot regeneration from nodal and axillary shoot tip explants. Among all the treatments BAP and Kn tested BAP at 4.0 mg/l and at 2.5 mg/l responds optimum in shoot regeneration from nodal explants and axillary shoot tip, respectively. Average number of sprouted shoots per explant and length of these shoots were determined after three weeks of explant inoculation. Axillaryshoot tip explants show best results of callus induction at 2 mg/l 2,4 diphenoxy acetic acid (2, 4-D) with a response percentage of 83.33%. Moreover, it was also noted that among the different concentrations of indole butyric acid (IBA) tested, IBA at the concentration of 1.5 mg/l resulted best in inducing the maximum number (12.66 ± 0.33) of in vitro adventitious roots from the callus. In this study combination of BAP + NAA (4.0 + 0.5) mg/l and 2 mg/l (2,4-D) resulted better in shoot induction and callus formation respectively in F. Krishnae.

1 illus, 3 tables, 25 ref

005527   NEGI Y, SHARMA S, SUTRADHAR N, ADHIKARI S (Biotechnology Dep, St. Edmund’s Coll, Shillong- 793 003, Email: samratadhikari@rediffmail.com) : A study on the differentiation of filamentous cyanobacterial isolates using DNA fingerprinting approach. Indian J Biotechnol 2019, 18(4), 337-45.

The present study investigates the implication of DNA fingerprinting in discrimination of class isolates collected from a spread of habitats in south west Khasi Hills district of Meghalaya, India. Five totally different primers i.e., M13, enterbacterial repetitive intergenic consensus (ERIC), short tandem repetitive repeats (STRRs) 1A & B and long tandem repetitive repeats (LTRRs), derived from repetitive sequences gift within the ordering of true bacteria, were utilized. The generated knowledge was then evaluated for fingerprinting exploitation procedure and unweighted pairgroup method of arithmetic average (UPGMA) based dendrograms. Clear and variable stripe patterns were obtained for every of the targeted genomic regions used, LTRR being the sole exception. The stripe patterns delineate the existence of 2 basic distinct populations of true bacteria among the 19 isolates. The UPGMA cluster analyses in addition, disclosed the presence of numerous lineages among the isolates. On examination the resolution capability of the four genomic regions studied, it had been seen that, with the exception of LTRR, all the opposite regions hold smart resolution and therefore the ability to differentiate between numerous class populations. The results of the current study indicate the incidence of genetic variants among the 19 morphologically identical isolates of filiform cyanobacteria. Additionally, the study additionally explicates the quality of DNA fingerprinting supplemented with UPGMA cluster analysis as a speedy, economical and value effective tool for differentiating among morphologically identical populations.

6 illus, 3 tables, 37 ref

005518   KUMAR D, BHAKTA S, ADHIKARY S P (Botanical Survey of India (WRC), Pune- 411001, Email: skmrbhakta@gmail.com) : Molecular phylogeny and spectral characteristics of selected soil crust cyanobacteria from tea gardens of Darjeeling (India). Indian J Biotechnol 2019, 18(4), 330-6.

Soil crust cyanobacteria from three tea gardens of Mirik, Kurseong and Darjeeling located at different altitudes of Darjeeling district (West Bengal, India) in Himalayas, were studied. A total of 16 cyanobacteria were isolated in pure culture and identified. Major taxa in all soil crusts from three locations were under the genus Calothrix. 16S rRNA sequencing of three dominant species of Calothrix were performed and the sequences were submitted to National Center for Biotechnology Information (NCBI) GenBank and the accession numbers were KF499103, KF499104 and KF499105. Phylogenetic analysis with other similar organisms isolated across the globe showed their relatedness with Calothrix species. Absorbance spectra of 90 % (v/v) methanolic extracts of the crusts and their major organisms in culture showed the presence of ultra-violet (UV) sunscreen pigment scytonemin and mycosporine like amino acids (MAAs).

5 illus, 1 table, 58 ref

005535   PRABHATH G P W A, SHUKLA S P, KUMAR K, NUWANSI K K T (Aquatic Environment and Health Management Div, ICAR- Central Institute of Fisheries Education, Andheri West, Mumbai, Email: spshukla@cife.edu.in) : Salinity mediated enhancement in protein and pigment content in spirulina (Arthrospira) platensis. Indian J Biotechnol 2019, 18(4), 323-9.

Spirulina (Arthrospira) platensis a filamentous multi cellular cyanobacterium utilized as animal feed and human food supplement. This cyanobacterium is also a rich source of value-added pigments with diverse applications in the food industry, pharmaceuticals and molecular biology. This report describes a procedure for enhancement of the yield of value-added pigment; c-phycocyanin (CPC) through the manipulation of salinity of the medium. The increase in salinity (sodium chloride-NaCl) from 0 to 3 g l^-1 of the prescribed growth medium resulted in a considerable increase (11.83%) in CPC content. Growth of the organism and the purity of the CPC were not affected at elevated salt concentration. However, a further increase in the salinity (NaCl) from 3 to 7 g l^-1 resulted in a  decline in the content (26.67%) and purity (31.13%) of CPC. Considering the enhanced growth and CPC content through the manipulation of the concentration of the salt NaCl in the medium, it is concluded that a moderate amount of the salt (3 g l^-1 NaCl) in the medium is a feasible and cost-effective approach for obtaining elevated protein biomass and enhanced the yield of the value-added pigment CCPS.

3 illus, 1 table, 22 ref

005538   RAJBONGSHI G, BARMAN N N, DAS S K, KHATOON E (Microbiology Dep, Assam Agricultural Univ, Guwahati- 781 022, Email: drgitika07@gmail.com) : Quantification of classical swine fever virus load by one-step TaqMan real-time RT-PCR assay. Indian J Biotechnol 2019, 18(4), 315-22.

A fluorogenic-probe hydrolysis (TaqMan) real-time reverse transcriptase polymerase chain reaction (RT-PCR) assay based on amplification of a 93 bp fragment from 5’un-translated region (5’UTR) of classical swine fever virus was used for detection and absolute quantification of the virus in clinical and tissue samples. For determining analytical sensitivity, an in vitro transcript RNA containing 5’UTR of classical swine fever virus (CSFV) strain Alfort187 from plasmid pCRXLV324-6 was used as a positive control and a standard for quantification of CSFV genomic RNA copies. The realtime quantitative RT-PCR (qRT-PCR) assay was used to assess the CSFV shedding from naturally infected pigs in whole blood, nasal swab and also in tissue samples. Used qRT-PCR was specific and sensitive as it could detect as low as 16.3 copies of CSFV genomic RNA. The assay was also reproducible as shown by satisfactory low intra-assay (0.80 % to 1.87 %) and inter-assay (1.00 % to 3.80 %) coefficient of variation with an efficiency of 102.3 % and R² of 0.993. Thus, the real-time qRT-PCR assay described here allows rapid, specific and sensitive laboratory detection and quantification of CSFV genomic RNA copies.

4 illus, 2 tables, 22 ref

005516   KOHLI P, MEHTA A, SINGH A, GUPTA R (Biotechnology Dep, Himachal Pradesh Univ, Shimla-171 005, Email: reenagupta_2001@yahoo.com) : Physicochemical properties of free and calcium alginate immobilized alkaline pectin lyase from Bacillus cereus. Indian J Biotechnol 2019, 18(4), 305-14.

Purified pectin lyase from Bacillus cereus was successfully immobilized in alginate beads with a high binding efficiency of 84.55 %. The optimal immobilization was achieved using 2.5 % (w/v) alginate concentration. Both free and immobilized enzyme showed optimum pH of 10.0 and temperatures of 40 and 45 °C respectively. Pectin lyase gave maximum activity at a substrate concentration of 0.5 % w/v for free and 0.75 % w/v for the immobilized enzyme and relatively similar Vmax values were obtained for both free (3.3 µmol/min) and immobilized pectin lyase (3.6 µmol/min). The Km for the immobilized pectin lyase (0.19 mg/ml) was slightly higher than that of the free (0.16 mg/ml) enzyme. The maximum inhibition of 50.2 % was observed in the presence of Hg²⁺ ion for free pectin lyase and immobilized enzyme showed maximum inhibition of 67.32 % in the presence of Na⁺ ion with statistically significant p-value (p < 0.05). Thermal stability was not significantly altered by immobilization. Thermal stability of free and immobilized pectin lyase was relatively similar at different temperatures. The immobilized pectin lyase retained almost 53 % of its original activity up to 7th cycle. Furthermore, during storage at 4 °C, immobilized pectin lyase retained relative activity of 79.77 % and free enzyme retained 63.63 % relative activity upto 35 days of storage, this indicated that the immobilization improved stability of the enzyme.

8 illus, 5 tables, 50 ref

005485   BANO N, SIDDIQUI S, AMIR M, ROOHI (Bioengineering Dep, Integral Univ, Lucknow, Uttar Pradesh, Email: roohi0607@gmail.com) : House of industrially important bioactive metabolites: A review on actinobacteria. Indian J Biotechnol 2019, 18(4), 293-304.

The actinomycetes are Gram-positive bacteria present in natural habitats, most profoundly in soil, freshwater, sea water and composts. Actinomycetes have high GC (guanine-cytosine) content and characterized by a complex life cycle. The morphology of actinomycetes looks like fungi and the reproduction takes place either by hyphae fragmentation or by spore production in the mycelium. Actinomycetes serves as a large pool for secondary metabolites production with potent biological activities and therapeutic properties like antifungal, antibiotics, enzymatic, plant growth promoting rhizobacteria (PGPR), in xenobiotics and bioremediation. These bioactive metabolites production can be enhanced using high-throughput fermentation and by combinatorial biosynthesis during optimization. Present research should be focused on the potential of less explored actinomycetes from extreme environments that can be utilized as sustainable herald for green biotechnology. In this review, habitat, life cycle and production optimization of actinomycetes in selective media are explained but article is emphasized on biotechnological potential of actinomycetes that are relevant for industries, agriculture and allied sectors.

3 illus, 4 tables, 119 ref

005546   SINGH A, BAJAR S (Environmental Sciences Dep, Central Univ of Jammu, Jammu- 181 143, Email: anitasaharan@gmail.com) : Optimization of cellulolytic enzyme production by thermophilic fungus Thermoascus aurantiacus using response surface methodology. Indian J Biochem Biophys 2019, 56(5), 399-403.

In the present study, thermophilic fungus Thermoascus aurantiacus, a local isolate, was used for production of cellulolytic enzymes using rice straw (RS) as substrate under submerged fermentation using Box-Behnken Design (BBD) of Response Surface Methodology (RSM). Themophilic fungus T. aurantiacus grew very well at 50-55°C temperature. BBD was used to study the influence of process parameters, pH (4-6), temperature (40-60°C) and substrate concentration (2-8 g/L) and their interactive effect on cellulase enzyme production. The R2 value 0.94 for Filter paper activity and 0.95 for carboxymethyl cellulase activity indicate that the model is appropriate and replicated to predict the effect of pH, temperature and substrate concentration on both enzyme activities. Under optimized conditions, 6.1 (U/gds) FPase activity and 28.2 (U/gds) CMCase activity was observed.

1 illus, 3 tables, 14 ref

005507   JAMWAL V L, GULFAM S, MANHAS R S, QAYUM A, KAPOOR N, CHOUHAN R, SINGH S K, CHAUBEY A, GANDHI S G (Plant Biotechnology Div, CSIR- Indian Institute of Integrative Medicine, Jammu-180 001, Email: sumit@iiim.ac.in) : Isolation, identification and bioactive potential of bacterial endophytes from Coleus. Indian J Biochem Biophys 2019, 56(5), 392-8.

Coleus (Lamiaceae) is a large and widespread genus comprising of species with diverse ethnobotanical uses. In the present study, bacterial endophytes were isolated from Coleus forskohlii and Coleus aromaticus. Endophytes are the microorganisms which reside within the plants without showing any harmful effect on its host. Diverse types of endophytes live symbiotically within almost all plants and in turn help the plant in a number of ways such as imparting resistance against biotic and abiotic stresses, producing compounds involved in attraction of pollinators, inducing the plant defense mechanisms, etc. The bacterial endophytes isolated in this study, were characterized by microscopic examination (using gram staining) and molecularly identified by sequencing the 16S rRNA. Extracts were prepared from endophytic biomass using solvents of different polarities (methanol, ethyl acetate and butanol) and were screened for their bioactive potential (in vitro cytotoxicity anti-microbial, and anti-oxidant activity). Scale-up of endophytes showing promising results is under process, which will help in isolation of pure compounds.

5 tables, 44 ref

005529   PARHI P, MULIK A, JADHAV V, YADAV A, SHOUCHE Y, BHADEKAR R (Microbial Biotechnology Dep, Bharati Vidyapeeth Deemed Univ, Pune- 411 046, Email: neeta.bhadekar@gmail.com) : Production and characterization of biosurfactant from Halomonas sp. BRI3. Indian J Biochem Biophys 2019, 56(5), 384-91.

In this report, the halotolerant bacterium Halomonas sp. BRI3 was studied for its biosurfactant (BS) producing ability. Effect of carbon source, nitrogen source, temperature, inoculum concentration and incubation on the production of BS was studied and maximum production (14 g/L) was obtained in modified medium containing 1 % glucose, yeast extract and ammonium chloride (0.25 % each) with 5 % inoculum concentration at 30 °C after 48 h, which was 2.8 fold higher as compared to original medium (5 g/L). Highest emulsification index (72 %) of crude BS was obtained with kerosene followed by n-hexane > crude oil > n-heptane > soybean oil > hexadecane > mustard oil > olive oil > sesame oil. Our hydrocarbon degradation experiments using crude oil and soybean oil revealed 40 % decrease in crude oil and 60 % decrease in soybean oil concentration after 50 days in presence of glucose, whereas, it was 20 % and 50 % in the absence of glucose, respectively. Based on TLC and FTIR analysis, the BS is chemically a glycolipo protein, demonstrated an significant antimicrobial and antiadhesive activity. We observed significant stability of BS over wide range of temperature (40 to 120 °C) and pH (5.0 to 11.0), suggesting its potential for application in food, pharmaceutical and cosmetics industries.

3 illus, 1 table, 37 ref

005500   GARCHA S, KANSAL R, GOSAL S K (Microbiology Dep, Punjab Agricultural Univ, Ludhiana- 141 004, Email: sgarcha@pau.edu) : Molasses growth medium for production of Rhizobium sp. based biofertilizer. Indian J Biochem Biophys 2019, 56(5), 378-83.

Rhizobium forms symbiotic relationship with leguminous crops and is recommended for use in various legumes. Rhizobium sp. fix atmospheric nitrogen and make it available to legumes through formation of root nodules. Rhizobium biofertilizer production is carried out mostly by using semi-synthetic microbiological medium which forms major expense of this activity. Successful commercial production of biofertilizer can be enhanced by use of natural substrates, as molasses, cheese whey, corn steep liquor, for bacterial biomass production. The present work centers around the use of sugarcane molasses as a source of fermentable sugars. It was supplemented with various organic/inorganic nitrogen sources, chemical compounds to increase biomass yield and to increase the shelf life of the product thus prepared. Compliance to Fertilizer Control Order specifications was demonstrated in wet lab analysis.

1 illus, 2 tables, 35 ref

005488   BHAT B, HABIB B, BHAGAT N, BAJAJ B K (Jammu Univ, Jammu- 180 006, Email: bajajbijenderk@gmail.com) : Cholesterol lowering and antioxidant potential of probiotic bacteria isolated from locally fermented milk product kalarei. Indian J Biochem Biophys 2019, 56(5), 363-72.

Probiotics due to their multifaceted health promoting attributes have gained immense research impetus in recent years. The current study reports the hypocholesterolemic potential of lactic acid bacteria (LAB) isolated from indigenous sources. LAB may use several mechanisms for lowering serum cholesterol level viz. cholesterol assimilation, bile salt deconjugation, and cholesterol adsorption on cell surface of live, resting and dead probiotic cells. Cholesterol lowering is generally a strain dependent phenomenon, and different LAB isolates exhibited varying level of hypocholesterolemic effects. Among the LAB isolates, K2 i.e. Enterococcus faecalis K2 showed the highest in vitro cholesterol lowering ability (82.32%), and strong bile salt deconjugation potential, and released about 128.43 µM/mL of cholic acid upon bile salt deconjugation. Furthermore, cholesterol removal by live, resting and dead E. faecalis K2 probiotic cells was shown to the extent of 72.46, 44.93, and 45.88%, respectively. Scanning electron microscopy displayed appreciable adherence of cholesterol on to the cellular surfaces of E. faecalis K2 cells. The antioxidant potential of the cell free cultural fluid of LAB isolates was quite variable. LAB isolate E. faecalis K2 showed appreciable DPPH radical scavenging activity (37.36%), hydroxyl radical scavenging ability (26.35%), and superoxide radical scavenging ability (42.67%). Most of the LAB probiotic isolates were susceptible to conventionally used antibiotics, and lacked biogenic amine producing ability and haemolytic activity. The probiotic isolate E. faecalis K2 may have potential for application for management of hypercholesterolemia related coronary heart diseases, however, after thorough in vivo investigation.

3 illus, 3 tables, 34 ref

005502   GUPTA P, NIPUNTA, DUTT K, SARAN S, SAXENA R K (Fermentation Technology Div, CSIR-Indian Institute of Integrative Medicine, Jammu- 180 001, Email: rksmicro@yahoo.co.in) : Binary immobilization: A newer approach for immobilizing lipase from a thermophilic sp of Thermomyces lanuginosus. Indian J Biochem Biophys 2019, 56(5), 358-62.

We report binary immobilization of Thermomyces lanuginosus lipase enzyme using chitosan as the support. This method of enzyme immobilization is better than cross-linked enzyme aggregate (CLEA) in terms of better enzyme recovery and separation. This method of immobilization resulted in an increase in the thermostability of the binary immobilized lipase as against the crude free enzyme. This preparation could be used for nearly 15 consecutive cycles with 80-100 % efficiency. Reusability of the immobilized enzyme makes it an economical alternative to the traditional way. Immobilized lipases in particular are a modern catalytic tool for various industrially significant reactions and applications.

2 illus, 2 tables, 21 ref