Research Journal of Biotechnology

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Vol. 15(7) July 2020

Tagging of novel thermosensitive genic male sterility gene in rice (Oryza sativa L.)

Sai Rekha K., Saraswathi R., Kumar M., Raveendran M. and Robin S.

Page No: 1-10

Abstract: The discovery of thermosensitive genic male sterility (TGMS) system has wide potential for creating new path to hybrid seed production technology in rice. It is imperative to study the mechanism of rice TGMS, a core component of two line hybrid development. In this study, TNAU 60S with male sterility was crossed with IET 21009, male fertile line and F2 population was developed. Six hundred and forty SSR makers covering twelve chromosomes of rice were screened between two bulk populations obtained from F2 representing fertile and sterile plants. Out of 640 markers, 181 markers were polymorphic between parents, out of which five markers viz. RM71, RM12665, RM6374, RM6942 and RM5015 located on chromosome 2 showed polymorphism between bulks.

In order to identify the precise location of thermosensitive genic male sterile (tms) gene, 86 markers were screened further and reduced to eleven. The tms gene was fine mapped on short arm of chromosome 2 at 24.3 cM between flanking markers RM12713 and RM12722. The identified tms locus can facilitate the screening of lines possessing tms gene. The markers linked to/flanking tms gene can be harnessed through marker assisted selection to accelerate two line hybrid rice breeding program.

RAPD and ISSR banding pattern of Globba marantina populations from Eastern India

Parida Reena and Nayak Sanghamitra

Page No: 11-14

Abstract:India is well known throughout the world as the land of aromatic plants, spices and traditional perfumes because of its favourable climatic conditions suitable for the development of these plants. The rhizomes of ginger plants are eaten raw or cooked as vegetables, used for flavouring food and in treatment of diseases. Medicinal plants are valued for curing various diseases which are depleting day by day in nature which require conservation to help in various ways to the pharmaceutical, cosmetic and other industries. Thereby the genetic study is very much essential to explore their molecular structure for producing high yielding varieties.

In the present work molecular characterization of Globba marantina from four different populations of Odisha using two molecular markers as Random Amplified Polymorphic DNA and Inter Simple Sequence Repeats has been done. A dendrogram was constructed through sequential agglomerative hierarchical and nested clustering, unweighted pair group method with arithmetic mean analysis and Jaccard’s similarity coefficient of combined markers which segregated these genotypes into two main clusters moreover showing the genetic similarity between the germplasm. Hence, the molecular analysis could be further used for identification of important novel gene present in Globba marantina which could be utilized for future crop improvement.

A common SELL gene haplotype associated with Cutaneous Leishmaniasis

Thanaa Ismael Jawad, Thuraya Aamer Habeeb, Fatima Abood Chaloob and Qasim Sharhan Al-Mayah

Page No: 15-20

Abstract:Allele, genotype and haplotype frequency of three single nucleotide polymorphisms in SELL gene (rs2205849 (-642C>T), rs2229569 (676C>T) and rs1131498 (616C>T)) were examined in a group of patients with cutaneous leishmaniasis (68 patients) in comparison with 70 apparently healthy subjects. Binary logistic regression and Chi square were used for calculation of odds ratio. None of these polymorphisms were associated with the incidence of cutaneous leishmaniasis.

However, haplotype analysis revealed that the haplotype block CCT from the three polymorphisms respectively was significantly associated with increase of the susceptibility to the disease (OR= 4.25, 95%CI=1.85-9.81). There was a strong linkage disequilibrium between these polymorphisms.

Simple and efficient DNA isolation protocol for small cardamom [Elettaria cardamomum (L.) Maton]

Ashokkumar K., Dhanya M.K., Murugan M., Haritha K.V., Nimisha M. and Surya R.

Page No: 21-25

Abstract: Small cardamom [Elettaria cardamomum (L.) Maton] is one of the most economically valuable spice crops since ancient period. The spice has been used in culinary, traditional as well as modern pharmaceutical applications. A remarkable variation is existing within the species which increases the chance for misidentiﬁcation and inadvertent duplication of varieties in many cardamom collections. Selection of parents without knowing its pedigree often leads to negative results. Therefore, information about germplasm diversity and genetic relationships among accessions or cultivars is of fundamental importance for breeding and the management of germplasm. Morphological characterization is influenced by various environmental factors. Hence, molecular characterization of cardamom accesions is essential identification of genotypes at molecular level. Currently, there is no efficient and cost effective DNA isolation protocol available due to presence of an diverse array of seondary metabolites, polyphenols and polysaccharides present in cardamom leaves. Therefore, this study aims to develop a DNA isolation protocol from the leaves of vazhukka cardamom accesions.

Emerging leaves of ten vazhukka type cardamom accesions were selected and used for genomic DNA isolation. A highly efficient DNA extraction protocol from freshly collected emerging leaves of cardamom was developed with modified cetyl trimethyl ammonium bromide (CTAB) method which is a collective stragegy of 2-mercaptoethanol, PVP (poly vinyl pyrrolidone), 1% sodium metabisulfide and 5M NaCl were used for removing polyphenols and polysaccharides. This protocol was pertinent to isolate genomic DNA from emerging leaf tissues of cardamom. Genomic DNA isolated by this method is highly suitable for characterization of cardamom accessions using molecular markers.

Chitosan Microparticles for Drug Delivery against Dental Caries

Khandelwal Amit Kumar, Bisht Shweta, Arya Rahul Kumar, Kharbanda Sumit and Singh Sujeet Pratap

Page No: 26-36

Abstract:Biofilm is developed by the bacteria with the production of extra polysaccharides components to become more resistant to the antibiotics. Dental caries is the breakdown of the tooth by the production of biofilm made by the bacteria. Particulate drug delivery systems have become important in experimental pharmaceutics and clinical medicine to transfer drug. Chitosan being highly hemocompatible, good swelling capacity and biopolymer is the efficient material that can be used as a material in drug delivery as it can cross link with the drug to be transformed. In the present work chitosan microparticles are prepared via sol-gel process to encapsulate rutin trihydrate and its action against biofilm is seen. The morphology of the particles prepared was analyzed using SEM resulting in nearly monodispersed spherical particles. The FTIR analysis of the particles has been done.

In the present work drug is post loaded into chitosan microparticles though impregnation method. The loading efficiency calculated was 32%. The in vitro drug release done in PBS buffer shows slow and sustained release of the drug from microparticles, the drug release profile showed that most of the drug was released within 7 hrs and then it starts decreasing after that. The hemocompatibility test was performed for chitosan microparticles and microparticles show it to be hemocomptible. The chitosan microparticles showed good swelling capacity. The synthesized microparticles are tested as a tooth filling material. The results showed that chitosan microparticles could be used as effective drug delivery carrier for drug against biofilm.

Bicarbonate supplementation improves biomass productivity in Scenedesmus obliquus cultivation

Matsudo M.C., Fukui-de-Sousa T. and Carvalho J.C.M.

Page No: 37-42

Abstract:Several studies have focused on the use of CO2 from alcoholic fermentation as carbon source in the cultivation of microalgae. However, considering the quantity of bioethanol produced in Brazil, for example, only part of this gas could be effectively used. In this sense, this inorganic carbon could be stored in the form of sodium bicarbonate and carbonate which can be used for supplementing microalgae growth medium.

In the present study, the green microalga Scenedesmus obliquus was cultivated in Erlenmeyer flasks and in bench-scale tubular photobioreactor with the addition of sodium bicarbonate (0.2 ~ 6.4 g.L-1). The supplementation of this inorganic carbon source allowed increasing biomass concentration in Erlenmeyer flasks. In tubular photobioreactor with better conditions for microalgal growth, this supplementation did not allow the increase in maximum biomass concentration, but permitted the growth in shorter time, increasing biomass productivity. Lipid content was not influenced ranging from 26.3 to 30.2 % and the fatty acids showed a satisfactory polyunsaturated fatty acids/saturated fatty acids ratio (PUFA/SFA= 1.0~1.1)

Morphological Analysis of T1 and T2 Transgenic Rice (Japonica Taipei 309) Progeny Plants harboring CCA1 Gene under the Green House

Devi Anita, Chaudhury Ashok and Sheoran Nayan Tara

Page No: 43-51

Abstract:Agrobacterium-mediated genetically transformed Japonica rice variety Taipei 309, T1 and T2 progeny plants for up-regulation and down-regulation of CCA1 (Circadian Clock Associated) gene. The experiment was conducted to study the expression of CCA1 Gene in Agrobacterium-mediated Genetically Transformed Japonica Rice. The experiment comprised mainly morphological characterizations. T1 transgenic progeny plants derived from constructs A, namely, A-17 and A-45 exhibited reduced number of tillers/panicles (6-7), reduced leaf length and leaf width (57cm to1cm) and decreased dry root-shoot mass A-17 to A-45 (19.3-22.1g) , WT (22.8g), panicle length (7.5cm), reduced starch content (by anthrone method) 5.21mg/ml compared to wild type plants. T1 and T2 progeny plants improved morphological characteristics as compared to wild type plants.

Present morphologically study demonstrates that endogenous repression of CCA1 gene resulted in improved morphological traits, increased number of tillers/panicle, plant height, leaf length and leaf width, panicle length, thousand seed weight, seed size, dry root-shoot mass, better starch content whereas over-expression leads to dwindling in morphological traits, decreased number of tillers/panicle, height, leaf length/breadth, starch content as compared to the wild type in T1 and T2 progeny plants. Analysis of transgenic plants and regulation of CCA1 gene is under progress and further helps in better understanding the role of CCA1 gene in plant growth, biomass and metabolism in future.

Unravelling of Osmotic genes involved in Drought tolerance in Backcross inbred lines of rice (Oryzasativa L.) cultivars

Baghyalakshmi K., Ramchander S., Raveendran M. and Jeyaprakash P.

Page No: 52-60

Abstract: Drought stress triggers a wide variety of plant responses, ranging from altered gene expression, cellular metabolism which changes the growth rates and grain yields. The primary physiological response of plant to stress begins with closure of stomata. This increases the accumulation of salts in the leaf cells and there is a variation in osmotic potential of the plants cell. The tolerant plants adjust the osmotic potential by synthesis of proteins like LEA, aquaporin, dehydrins and maintain the turgidity of the cell. In this study an attempt was made to screen the Backcross Inbred Lines of rice obtained from the cross between IR64 X Apo which carried three mega QTL viz. CB 229 (qDTY2.2, qDTY3.1 and qDTY8.1), CB 193-1 (qDTY3.1 and qDTY8.1), CB 193-2 (qDTY2.2 and qDTY8.1) and CB 193-3 (qDTY2.2 and qDTY8.1) along with the parents to know the profound expression of QTLs under severe drought condition.

The BIL CB229 with high osmotic adjustment during drought stress along with the parents were grown under controlled condition and RNA was extracted to study the genes involved in the osmotic homeostasis and accumulation of compatible solutes. About 15 genes were unregulated in tolerant parent Apo, 13 genes were commonly upregulated in Apo and CB-229 and 31 genes in CB-229 alone. These genes can be engineered further to develop lines which show enhanced tolerance to drought environment.

Bam, Sex lethal, Ago1 and Brat multiple protein complex mediates negative regulation of dMyc mRNA in S2 cells of Drosophila melanogaster

Malik Sumira

Page No: 61-69

Abstract: The differentiation factor Bag of Marbles (Bam) is required for differentiation of Germ line stem cell in the Drosophila melanogaster ovary showing its high expression in differentiating cystoblasts and dividing cysts. Diminutive (dMyc), the human c-myc oncogene and the homolog of an important regulator of cell growth are highly expressed in self-renewing stem cells but its expression is downregulated in differentiating cystoblasts and 16-cell cysts. The molecular mechanisms responsible for such downregulation of dMyc mRNA in the cystoblasts and 16-cell cysts stage needed to be addressed.

The current finding signifies that Bam forms multiple complex with Ago 1, a microRNA pathway component, Brat a TRIM–NHL domain containing factor and a RNA-binding protein sex lethal which is specific to polyuridines stretches. The multiple complex of Bam, Sex lethal, Ago1 and Brat represses dMyc mRNA through 401-704 bp region of dMyc 3’UTR consisting of the six microRNA sites containing differentiating daughter cells and cystoblasts.

Complete genome sequence of free living Pseudomonas aeruginosa 6A (bc4): Profiling of genes involved in cellular metabolism

Marathe R.J., Phatake Y.B. and Sonawane A.M.

Page No: 70-77

Abstract:The aim of the present study was to analyze whole genome sequence of P. aeruginosa strain 6A (bc4) isolated from rhizospheric soil sample for studding various genes involved in cellular metabolism and in bio remediation activity. Pseudomonas aeruginosa 6A (bc4) strain previously isolated and characterized from soil sample was used in this study. Genomic DNA of the selected organism was isolated by CTAB method and characterized by Qubit 2.0 Fluorometer. The paired-end sequencing library was prepared using Illumina Tru Seq Nano DNA HT Library preparation kit. The next generation sequencing for WGS sample was performed using paired end (PE) 2x150 bp library on Illumina platform. Genomic data revealed presence of total 5649 genes and 149 contigs. The GO (Gene Ontology) annotation classified 3593 genes under different molecular functions, 3804 genes into biological process and 1654 were classified into cellular component category. KEGG Automatic Annotation Server (KAAS) was used to analyze the genes belonging to metabolic pathways. 2516 genes were found to be involved in various metabolic pathways and high percentages of genes were predicted as regulatory genes.

The complexity of the 6A (bc4) genome suggests that the bacterium is capable of adaptation into diverse environmental conditions and is able to degrade different pesticides and complex molecules. Analysis of the complete genome sequence of P. aeruginosa reveals many clues regarding the versatility of this organism. It also has broad capabilities to transport, metabolize and grow on organic substances and numerous iron-siderophore uptake systems. Extensions of the work described here have the potential to produce a detailed model for genetic variation in P. aeruginosa with more emphasis on the genes function in bioremediation and pesticide degradation.

Prediction and design of zinc finger target sites for an important human regulatory region (locus control region)

Sharma B. Sharan and Verma Ramtej J.

Page No: 78-82

Abstract:In order to understand the regulation of a gene, it is important to understand the mechanism of DNA-protein interaction at the molecular level. Recognition of zinc finger domains is one approach to discern the target sites of DNA-protein interactions. Utilizing this information, zinc finger proteins (ZFPs), most common DNA binding proteins in mammals, can be synthetically designed and engineered for the modulation of endogenous genes expression and for targeted genome engineering. Here, we discovered zinc finger target sites in an important regulatory DNA sequence tyrosinase 5' upstream regulatory sequence (TYR 5'URS) using online bioinformatics tool ‘Zinc Finger Tools’ (ZF Tools). TYR 5'URS acts as a transcriptional cis regulatory/locus control region (LCR) of a multifactorial tyrosinase (TYR) gene. A total of 79 top scoring zinc finger target sites could be detected in TYR 5'URS. Top score of the identified target sites ranged from 40.87 to 61.01.

Further, we generated protein coding sequences for the top scoring target sites which contained canonical amino acid linker between the zinc finger modules. Amino acid sequences of designed ZFPs, when subjected to identity analysis, did not correspond significantly to any known human protein indicating the specificity of the zinc fingers for the target sites. Information and data generated in this work will be further helpful to design synthetic ZFPs and to assemble the zinc finger-DNA binding domain coding sequence to target TYR 5'URS. Designing ZFPs specific to TYR gene regulatory elements will allow us to explore and understand the mutation spectrum of this gene in a population. This will further pave the way to therapeutic interventions.

Physiological and Biochemical characterization of Sclerotium rolfsii with respect to its infection on ground nut

Tatmiya Ritisha N., Ambalam Padma S. and Tomar Rukam S.

Page No: 83-88

Abstract:The physiological and biochemical changes in groundnut (Arachis hypogaea L.) during the Sclerotium rolfsii infection were carried out on selected groundnut genotypes viz. CS-319, CS-19, JAL-42, GG-20 and TG-37. The present study involved collection of leaf tissues before and after 3 days of inoculation of S. rolfsii for physiological and biochemical analysis. Enzyme activities were also ascertained due to stem rot infection in groundnut.

The physiological parameters such as chlorophyll a, chlorophyll b and total chlorophyll reduction were observed in the infected plant as compared with the control plant. The biochemical parameters like total phenol, free amino acid and protein content increased while total sugar content decreased in infected plant. The catalase and peroxidase activity was noted higher with significant correlation in infected plant.

Assessment of molecular diversity in sesame (Sesamum indicum L.)

Pavani K., Lal Ahamed M., Ramana J.V. and Sirisha A.B.M.

Page No: 89-97

Abstract:The molecular characterization of 30 sesame genotypes with 50 SSR primers revealed the use of 45 primers for characterization as they produced clear banding pattern. Only seven primer pairs showed polymorphism and total number of bands produced were 191. The number of alleles per locus was varied from 2 to 6 with an average of 3.57 alleles per locus. The size of the scoring bands ranged from 120 to 300 bp. PIC values of primer pairs ranged from 0.28 (SM-10-118) to 0.80 (SSR-217) with an average value of 0.47. The number of effective alleles (Ne) ranged from 1.38 to 4.86 with an average of 2.49. Average expected heterozygosity (He) values ranged from 0.27 to 0.79 with an average of 0.53.

Average Shannon’s diversity index (Ic) was 0.95 with the range of 0.40 to 1.65. These allelic variation parameters indicated the use of their exploitation in diversity studies of sesame. Dendrogram analysis grouped 30 genotypes into 4 clusters and one clade exhibiting genetic similarity coefficient values of 0 to 1.0. The genotypes of the clusters II (YLM-153, YLM-154 and YLM-155) and IV (MLTS-1, MLTS-2, MLTS-3, MLTS-4 and MLTS-5) had high values of dissimilarity of 1 indicating the presence of wide genetic base in the material and their use in breeding programmes.

Comparative analysis of enzymes production by Aspergillus niger using Pretreated Albizzia lebbeck fruit pods

Pingili Mamatha, Marla Shailaja Raj and Raparla Ramakrishna

Page No: 98-104

Abstract:In the present research production of endoglucanase, Fpase and xylanase enzymes was studied on different pretreated Albizzia lebbeck fruit pods as inducer to illustrate the relationship between enzyme production and various pretreatment methods. Physicochemical modifications of untreated and pretreated biomass were carried out by FTIR and SEM analysis along with monitoring the corresponding compositions. Solid state fermentation of acid and alkali treated fruit pods were carried out by Aspergillus niger. Aspergillus niger was inoculated separately on to steam exploded, acid and alkali pretreated fruit powder supplemented with basal media. Endoglucanase, Fpase and xylanase enzyme activities were measured for all the systems (steam exploded, acid and alkali pretreated fermentation systems).

Of all, alkali pretreatment showed best results with 3.682U/ml of Endoglucanase activity, 2.604 FPU/ml of Fpase activity and 8.029 U/ml of Xylanase activity. Xylanase activity was found to be greater for acid pretreated fruit pods (9.291 U/ml) on 2nd day of fermentation. Pretreatment of Albizzia lebbeck fruit pods with NaOH can be considered for enhanced production of cellulases and Xylanase enzymes by Aspergillus niger.

Cytotoxicity of Salacia oblonga extracts against cancer and normal cells and isolation of bioactive compounds

Anjaneyulu Musini, Bhargavi Pokala, Zahoorullah S.M.D. and Archana Giri

Page No: 105-109

Abstract:Salacia oblonga (Wall.) is a woody plant found in the forest of India and Sri Lanka. It possesses different pharmacological properties. The roots and stems of Salacia oblonga have been used widely in ayurveda and traditional Indian medicine for the treatment for diabetes. In the present study, phytochemicals were extracted in methanol solvents. Cytotoxicity of S. oblonga aerial and root extracts was evaluated against different cancer cell lines like human metastatic ovarian cancer cells (SKOV3), human cervical cancer cells (HeLa), breast cancer cell lines (MDA MB-231) and normal Chinese hamster ovary cells and lymphocytes.

Cancer cell lines were treated with various concentrations of the extracts ranging from 6-600 µg/ml for 24 h in order to evaluate the cytotoxic effect by using the MTT assay. Extracts showed low IC50 values against different cancer cell lines and aerial extracts did not display any cytotoxicity towards normal cell lines. Separate and identify the compounds by using different chromatographic techniques. Bioactive compounds were identified from the LC-MS analysis. Our study highlighted the medicinal importance of Salacia oblonga Wall.

Biotechnological interventions for sustainable plant secondary metabolite (amarogentin) production under the harsh environmental conditions of Himachal Pradesh: an ecofriendly approach

Kumari Garima and Kanwar Kamlesh

Page No: 110-119

Abstract:Swertia chirayita Buch.-Hams. ex Wall plants were micropropagated with hundred per cent survival on field transfer. Quality of micropropagated plants checked through use of modern validated methods revealed the presence of amarogentin with no significant difference among both seedling- stages and micropropagation stages authenticate the material of study. Highest percent amarogentin content was noticed at flowering stage (0.4593%) in seedling raised plants. Micropropagated plants were observed with maximum per cent amarogentin content at five months old stage (0.3647%). Percent amarogentin content was more at three, five and seven months old stages for micropropagated plants in comparison to seedling grown plants. Presence of amarogentin may increase pharmaceutical value of micropropagated plants. Further, use of two types of molecular markers RAPD and ISSRs for assessment of genetic stability of micropropagated plants was raised through indirect organogenesis resulting in 85% and 90% of similarity respectively between mother plant and micropropagated plants.

Dendrograms were derived on UPGMA clustering analysis using similarity coefficient of RAPD and ISSR markers separated micropropagated plants and mother plants into two separate clusters at maximum similarity value showing greater homogenity among them. The similarity coefficient value ranged from 0.80 to 0.95 in RAPDs study and 0.88 to 0.97 using ISSRs. Maximum per cent similarity was observed among micropropagated plants. Further, S4 and S5 were found highest, 97% similar in both RAPDs and ISSRs.

Virulence gene profile and biofilm formation ability of Staphylococcus isolated from clinical and subclinical bovine mastitis in Bihar

Jha Vikash Kumar, Kumar Rohit, Singh Rahul, Singh Ravi Shankar, Roy Prabhat Kumar and Thakur Dharamsheela

Page No: 120-126

Abstract:Mastitis is the one of the most prevalent and costly disease of dairy industry. Mastitis is multi etiological and complex in nature. Among bacterial mastitis, Staphylococcal mastitis is the predominant type. Pathogenic potential of Staphylococcus depends on the numbers and combination of virulence genes and also on the geographical distribution. This study was conducted in order to screen and identify the major virulence factors of coagulase positive and coagulase negative Staphylococcus, isolated from clinical and subclinical bovine mastitis milk samples collected from Bihar state of India so that they could act as immunogenic target for vaccine development.

Genomic DNA was isolated from the staphylococcal isolates and was screened by PCR for the presence of genes associated with biofilm formation (bap, icaA and icaD); genes encoding microbial surface components recognizing adhesive matrix molecules (MSCRAMMs; cna, ebpS, eno, fib, fnbA, fnbB, clfA and clfB) and tsst-1 encoding superantigenic toxin. All the isolates were found positive for eno (100%) followed by clfA and fib in 84%; fnbA and tsst-1 in 60%, clfB (52%), cna (44%), fnbB (40%), ebps (16%), bap (16%), icaA(10%) and icaD (8%) of the isolates. Biofilm formation ability of the isolates was also tested by microtiter plate method. 66% of the isolates were found to be biofilm producers with different production level. Out of the biofilm positive isolates, 10% were strong, 32% moderate and 24% were weak biofilm formers whereas 34% of isolates did not form biofilm. The most prevalent virulence factors were thus identified as eno, clfA, fib and tsst-1 which could be used as potential immunogenic target for vaccine development against staphylococcal mastitis.

Study of Identification of Hemoglobin Variants and their Clinical Implications in Tribal Population of Sirohi District

Parmar Sonika, Toteja G.S. and Chauhan Ritu

Page No: 127-130

Abstract:Hemoglobin (Hb) is a heterotetramer composed of α-like and β-like globin subunits each bound to a heme prosthetic group. Sickle cell disease affects the hemoglobin of the person as the shape of RBC becomes cresent shape, therefore this may lead to various acute and chronic complications, several of which are potentially lethal. The study was conducted with the aim to identify different haemoglobin variants in the region and also to find association between clinical manifestation and haemoglobin variants. A cross- sectional study was carried out in tribal population of Sirohi district of Rajasthan to identify the different Hb variants in the region and their clinical implications. Cluster randomized sampling was used to screen the population.

Total 38 (25 SCT and 13 SCD) subjects were assessed for clinical symptoms. Fever were common symptoms in both SCD and SCT cases, 84.61% of subjects reported fever in SCD patients whereas 20% subjects of SCT reported fever followed by weakness (69.23%) in SCD subjects and 12% in SCT subjects. Pain crisis was also common among both group, 69.23% reported pain crisis in SCD subjects while 12% subjects reported pain crisis in SCT. Fever and Anaemia are most common problems found in both Sickle Cell Disease subjects as well as in Sickle Cell Trait subject. Fever, Weakness, Pain crisis, Anaemia Jaundice, Cardiac Complication Renal Complication Leg Ulcer are very commonly found in the subject having SCD and SCT.

Medicinal Value of Secondary Metabolites of Pines grown in Himalayan Region of India

Bhardwaj Kanchan, Bhardwaj Prerna and Kaur Simran

Page No: 131-140

Abstract:This review focuses on the geographical distribution of Pinus species of Indian Himalayan Region (IHR) with special reference to the pure and mixed stands; also throws light on the medicinal importance of the secondary metabolites. The Indian Himalayan Region has high rich varieties of plants which have their medicinal properties to cure different ailments. Since prehistoric time, people rely on these plants as herbal medicines against the treatment of various diseases. Increasing demands of these medicinal drugs and lower side effect, nowadays the pharmaceutical industries mostly depend on the plant products.

Among different Himalayan species, coniferous forest plays a vital role in meeting the timber needs as well as commercial purposes. Among conifers, pine tree forests are of great economic and medicinal importance. Five pine species which are indigenously found in the Himalayas in India are Pinus roxburghii, Pinus wallichiana, Pinus gerardiana, Pinus kesia and Pinus merkusii. Different types of secondary metabolites are produced by them such as polyphenols, alkaloids, flavonoids, tannins, terpenes and ascorbic acids and are effective against diseases such as cancers, diabetes, asthma, liver and kidney disorders, cardiovascular-related problems and many other bacterial and fungal diseases. This review also recommended the role of different States and Central government along with many research institutes for conservation and development of Pinus species.

Lipases with preferred thermo-tolerance in Food Industry

Govada Vayunandana Rao and Kota Sobha

Page No: 141-150

Abstract:Lipases are defined as ‘esterases’ of carboxylic acid because of their explicitness for carboxylic acid ester bonds. In general, these enzymes can be of bacterial, fungal and plant (seeds) origin with catalytic property variations. Extensive use of lipases in industries is due to the exhibition of stereo, regio and substrate specificity and thermostability. This review provides a gist of lipase applications and from the synthesized gist it was evident that lipases are extensively used in dairy and flavour industries. Emphasis was laid on applications of thermostable lipases especially in food industry which provided novel strategies as a substitute for chemical flavours in food industry. Synthetic chemical flavours usually improve the food taste but they show certain devastating effects if they are not within acceptable daily intakes.

In order to beat the chemical flavour toxicity, an attempt is made here to tackle these issues with thermostable lipases produced flavours with evidence of some research in this direction. Finally, this review aims at providing the need to research more natural lipases and standardize their usage in food industry.

Endophytic microorganisms: colonization, plant-microbe interaction, diversity and their Bioprospecting

Barman Dina and Dkhar Mamtaj S.

Page No: 151-179

Abstract:Endophytic microorganism, belonging to bacteria and fungi are found in the intercellular and intracellular spaces of plant parts forming a wide range of relationship. Almost all the plant species of the earth mainly belonging to crop and medicinal plants are harboured by numerous numbers of endophytic bacteria and fungi. They are mainly originated from rhizosphere, phyllosphere and spermosphere. They represent the unexplored reservoir of an impressive array of secondary metabolites including antibiotics, antitumor agents, plant growth promoters and enzymes for exploitation in pharmaceutical, agriculture and industry. However, the methods of surface sterilization, isolating media and characterization are differing among the bacterial and fungal endophytes.

Additionally, the investigation of the indigenous population and monitoring the colonization of endophyte in the host plant are also important criterion. Keeping in mind, this review focuses on different isolation methods, surface sterilization and diversity of endophytic bacteria and fungi with their potential biotechnological application. We also highlighted the mechanism behind the interaction of endophytes with host plant species.