Research Journal of Biotechnology

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Research Journal of Biotechnology

Expression Analysis of DREB2A Transcriptional Factor under drought stresses in rice (Oryza sativa L.)

Yadav Ashwani, Sharma Anubhuti, Kumar Ashwani, Yadav Renu, Misra J.P., Kumar P., Singh D. and Kumar Rajendra*

Rice (Oryza sativa L.) is highly sensitive to drought and the effect of drought may vary with different genotypes and development stages. Drought and other abiotic stresses negatively impact plant growth and productivity of crops. The DREB2A transcription factors have important roles in abiotic stress-responsive signalling. Transcription factors (TFs) are master regulators of gene expression. The transcriptomes from leaf at different stress stages were comparatively analyzed and combined with bioinformatics for exploring drought stress related cis-elements. We investigated the gene expression response of four selected genotypes of rice Nagina-22 and Annada (drought tolerant genotypes) & Pusa Basmati-1 and Basmati-370 (drought susceptible genotypes) up to 7 days of drought stress. In order to study DREB2s, the authors analyzed the expression patterns of these genes under drought stress conditions and obtained its full-length cDNA sequence by bioinformatics and laboratory bench-work.

Based on the sequence, cDNAs of rice were amplified with RT-PCR and BLASTN analysis based on deduced amino acid sequences was done that revealed significant sequence similarity to DREB2A proteins belonging to diverse families of plant species. Results showed that the expression pattern in Annada and Nagina-22 increased up to 5th day of dehydration and showed slight decreased expression on 7th day. However in Basmati-370 and Pusa Basmati-1, DREB 2A level decreased on 5th day and slightly increased on 7th day of dehydration stress. Our analysis highlights changes in the expression configuration of DREB 2A gene and depicts a limited but highly specific drought responsive stage at 5th day of degradation.

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Species discrimination of the genus Citrus and the hybridization analysis

Sun Yan-Lin*, Jin Cheng-Wu and Hong Soon-Kwan*

The genus Citrus (Aurantioideae, Rutaceae) is the most important fruit crops all around the world. As many cultivated and weedy species/varieties exist in this genus, botanists hold different opinions on the taxonomy and phylogeny of the genus Citrus all the time. To evaluate the taxonomy and phylogenetic relationship of Citrus species, we collected 22 Citrus species and evaluated the genetic diversity and phylogeny based on the genetic variation of the ribosomal RNA (rRNA) 5S gene sequences. The 5S DNA alignment showed high nucleotide variations and sequence length difference among investigated species. In the rRNA 5S gene, four nucleotide sites were found to be easily nucleotide-substituted and hybridization-reflective.

Some Citrus species, including C. kinokuni, C. unshiu, C. sinensis, C. leiocarpa, C. tagerina andC. hybrid are found to be hybridization-susceptible species. The analysis result of genetic distance suggested that C. clemntina, C. hybrid, C. leiocarpa, C. nippokoreana, C. kinokuni and C. tagerina showed low values with each other and divided into one group in the rRNA 5S phylogenetic tree. Others were grouped together in the 77% similarity criterion. According to six-tribe discrimination method, the clading seen from the 5S phylogenetic tree suggested that the species belonging to Sinocitrus tribe showed higher sequence variation than other tribes. This work provides more sequence evidences for Citrus species identification and helps us further understand the phylogenetic relationships and species identification of the genus Citrus. The results would help citrus breeding and germplasm conservation programs.

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An in silico approach to resolve synovial inflammation by inhibiting Wnt signalling via post-translational modifications: phosphorylation and S-nitrosylation

Verma Mahendra Kumar and Kota Sobha*

Rheumatoid arthritis (RA) is a chronic progressive autoimmune disorder with complex etiology which is not explored completely. RA affects primarily long bone joints, but recent findings have suggested it is spread to other joints too. The immune system centrally involves in the initiation and progression of the disease. The immune activity results in the production of various inflammatory mediators that start acute inflammation of synovial joints, a defensive mechanism to encounter the causes. The problem arises only when acute inflammation fails to resolve and leads to chronic. At this stage, inflammatory mediators stimulate other metabolic pathways resulting in tissue damage. The drugs available for clinical applications in RA offer only a symptomatic relief and fail to resolve inflammation. It may be because of a complex RA etiology and involvement of several metabolic players in the initiation and progression of the disease.

Fibroblast-like synoviocytes (FLS) were reported an important player in the progression of RA and are under control of Wnt signalling pathway. Here, we propose inhibition of Wnt signalling pathway and blocking the activity of Fibroblast-like synoviocytes (FLS) to resolve synovial inflammation. A control over Fibroblast-like synoviocytes (FLS) also provides substantial relief in the production of other inflammatory mediators in the synovial cavity in the course of RA. We aimed an in silico approach to block Wnt signaling using two post translational modifications - phosphorylation and S-nitrosylation of Frizzled proteins (Fz), a family of G protein-coupled receptor proteins which in turn will take control over Fibroblast-like Synoviocytes (FLS) mediated inflammation of synovial joints.

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Genetic diversity and relationships among rice accessions (Oryza sativa L.) of cultivated and weedy types using CACTA-TD and AFLP markers

Ramekar Rahul Vasudeo, Farooqi Muhammad Qudrat Ullah, Sa Kyu Jin, Park Kyong-Cheul and Lee Ju Kyong*

For understanding the genetic diversity and genetic relationship between cultivated and weedy types, we evaluated genetic variation of 80 accessions of rice (O.sativa). This included 42 cultivated accessions and 38 weedy accessions with the help of AFLP and CACTA-TD. A total of 542 loci were analyzed (255 for AFLP and 287 for CACTA-TD) of which AFLP markers exhibited 75% of polymorphism and transposon based CACTA-TD markers exhibited 93% of polymorphism. The average genetic diversity value for all 80 accessions using AFLP markers was 0.226(Cultivated – 0.210; Weedy 0.241) and based on CACTA-TD markers was 0.281 (Cultivated – 0.294; Weedy 0.269).

A UPGMA phylogenetic tree revealed three major groups for both the marker system. The average polymorphic content value obtained with AFLP and CACTA-TD markers were 0.21 and 0.232, Effective multiplex ratio (AFLP – 47.50; CACTA-TD– 66.75), Marker Index (AFLP – 9.94; CACTA-TD– 21.13) and Resolving power (AFLP – 19.53; CACTA-TD– 34.62) indicated that the CACTA-TD markers were relatively efficient than AFLP markers.

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Effects of Exogenous SA (Salicylic Acid) on Chilling Resistance of Tibetan Medicine Lamiophlomis rotata 1

Ziping Cai, Yuan Chen*, Hongxia Wang, Guoxiang Wang and Ashraf Muhammad Aqeel

SA with different contents is sprayed on two pairs of leaf-age Lamiophlomis rotata seedlings which are then quickly placed in the environment of 40C for 48h low-temperature handling. With the control group at 250C, the changes of seedling leaves’ electrolytic leakage, cytomembrane’s stability, autioxidant enzyme activities, peroxidization of cytomembrane and other indicators are measured after the handling, aiming to reveal the effects of exogenous SA on improving the chilling resistance of seedling Lamiophlomis rotata so as to provide theoretical references for the artificial culture of Lamiophlomis rotata .

Results indicate that Lamiophlomis rotata seedling can adapt to the low temperature of ℃ to a certain degree. The spraying of 5.0 mmoL.L-1SA can significantly induce and enhance the activity of SOD and POD in Lamiophlomis rotata seedling leaves, reduce peroxidization of cytomembrane, strengthen the stability of cell membrane system, effectively improve the chilling resistance of Lamiophlomis rotata seedlings, relieve the harms of rapid freezing on seedlings and lay a sound foundation for the recovery and growth.

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Biosynthesis of silver nanoparticles using fungus Penicillium brevicompactum and evaluation of their anti-bacterial activity against some human pathogens

Solanki B. D.*, Ramani H. R., Garaniya N. H. and Parmar D. V.

Nanoparticles synthesis with biological material is extremely appealing for a variety of commercial, scientific and health applications. The current study reports biosynthesis of silver nanoparticles (Ag-NPs) with Penicillium brevicompactum. Ag-NPs were rapidly synthesized and their characterizations were done by different analytical techniques like UV- Vis spectra, TEM, SEM-EDAX, FTIR and XRD. UV–visible spectra of Ag-NPs showed an absorption peak at 420 nm for silver corresponding to the surface plasmon resonance of silver.

The TEM and SEM-EDAX studies revealed the formation of mono-dispersed spherical metallic silver nanoparticles with size ranging from 6.28 – 15.12 ± 0.8 nm while FTIR spectroscopy confirmed the presence of extracellular proteins on surface, their XRD spectrum confirmed the formation of metallic silver nanoparticles. We also examined antibacterial assay of silver nanoparticles against clinically isolated human pathogens: Staphylococcus aureus, Pseudomonas aeruginosa and Escherichia coli and the results obtained showed potential of Ag-NPs as antibacterial agents in the field of medical sciences.

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Genetic Crossover Key generation mechanism for cryptographic applications

Ruhan Bevi* and Malarvizhi S.

This paper discusses the key generation methods for cryptographic applications using genetic reproduction. The methodology of seed selection, crossover points and mutations is formulated using the Linear Congruential Generator (LCG) recurrence relations. A chromosome is represented with 8 binary encoded genes and a set of 10 chromosomes forms a generation. Spin ring crossover operation (SRC) is induced in the reproductive stage of the parental chromosomes. The recurrence relation is iterated for 100 generations G1 to G100 and the deviation between the generations G1- G100 is computed. The keys for the cryptosystem derived from the generated population are tested for randomness and deployed for cryptographic key generation methods.

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Potential characterization and antimicrobial applications of newly bio-synthesized silver and copper nanoparticles using the novel marine-derived fungus Alternaria tenuissima KM651985

Abd El Aty Abeer A.* and Ammar Hala A.

Nanotechnology needs to develop alternative method to chemical synthesis. One such method is biological synthesis employing microorganisms, that is reliable, non-toxic, clean and ecofriendly. In this paper the novel marine-derived fungus Alternaria tenuissima KM651985 which is able to produce lignocellulytic enzymes, has been subjected to extracellular biosynthesis of silver and copper nanoparticles (AgNps and CuNps). The biosynthesized nanoparticles were studied and characterized by UV-visible spectroscopy, Fourier transform infrared spectroscopy (FTIR) and transmission electron microscopy (TEM).

Our measurements have revealed the formation of spherical, well-dispersed silver nanoparticles with size ranging between 2 to 15 nm and maximum absorbance peak of 430 nm at optimum conditions of 4 mM AgNO3 solution incubated with fungal extract for 72h. Mixing fungal extract with 2 mM CuSO4. 5H2O for 72h was preferred for the biosynthesis of CuNps (particle size in range from 16-51 nm) with characteristic peak at 460 nm. The antimicrobial studies of biosynthesized nanoparticles display that AgNps and CuNps have attention opposing both classes of bacteria, fungi and yeasts but alongside disparate degree. From this study we have concluded that the marine-derived fungus Alternaria tenuissima KM651985 is a novel good candidate for biosynthesis of AgNps and CuNps. AgNPs showed strong antimicrobial activity as compared with CuNps and received considerable attention as antimicrobial additives that can be requested in health-related and manufacturing products.

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Afterglow corona discharge air plasma (ACDAP) for microbial decontamination of shell eggs: Effect on physicochemical properties

Puligundla Pradeep, Choi Soee and Mok Chulkyoon*

Non-chemical disinfection methods are gaining popularity in egg sanitation. In this study, afterglow corona discharge air plasma (ACDAP) was used for microbial decontamination of shell eggs. ACDAP was generated at an output voltage level of 20 kV DC and at a frequency of 58 kHz. Microbial contaminants including aerobic bacteria molds and yeasts and other pathogenic bacteria, namely Staphylococcus aureus were detected on shell surface. Upon the plasma treatment for 12 h, 1.4 log (96%)-1.7 log (98%) reductions of the detected microorganisms were observed. The microbial inactivation pattern fitted well to Singh-Heldman model or pseudo-first-order kinetics.

No statistically significant (p>0.05) changes in physicochemical (weight, Yolk index, Haugh units, pH) and sensory characteristics (appearance, aroma, flavor, texture) of eggs were observed due to ACDAP treatment. In addition, improved (p<0.05) storage quality of eggs was observed upon the plasma treatment. The results of this study demonstrate that ACDAP can reduce the overall microbial load including pathogenic bacteria on eggshell surface. The plasma treatment can also help maintain storage quality of eggs for 3 weeks at 25˚C without compromising their physicochemical and sensory properties.

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Cosmeceutical Activities of Lactobacillus acidophilus HK-9 in the Human Skin

Lee Hyeon Yong

A new microorganism, Lactobacillus acidophilus was isolated from human skin, especially in the hands of women by tree analysis of the amplified 16S rRNA gene. Its morphology was observed as a rod-typelactobacillus through scanning electron microscopy and named Lactobacillus acidophilus HK-9 (KCCM11789P) by the Korea Culture Center of Microorganisms.

Because the isolated bacterium was derived from human skin, its cosmeceutical activities were observed revealing a relatively high antioxidant effect as 14.95% of DPPH free radical scavenging ability and also 18.26% of inhibiting tyrosinase activity as well 55.10% of inhibiting melanin synthesis. It was also confirmed that these cosmeceutical activities were similar to or even better than those from other natural herbs which can strongly indicate that a newly isolated Lactobacillus acidophilus HK-9 could be used as an alternative cosmeceutical whitening agent.

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