Research Journal of Biotechnology

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Identification of Long noncoding RNA and mRNA in Yak coat color using RNAseq

Guangxin E., Tianwu An and Xiaolin Luo

In this study, we used high-throughput technology to provide the first transcriptome dataset for differentially expressed genes in mixed pools of dermis tissue from black and white coat yak to research on the possible molecular mechanisms of yak coat pigment. In this study, 92,636,002 and 95,917,842 clear reads were generated through Illumina paired-end sequencing. A total of 2,075 differentially expressed genes (DEGs) were identified including 835 upregulated and 1,240 downregulated genes in the mixed pools of white coat yak compared with the mixed pools of black coat yak. Regards to long noncoding RNAs (lncRNAs), 59 significantly differentially expressed lncRNA (DELs) were found including 26 up-regulated and 33 down-regulated. These DEGs were related to 60 GO categories and enriched in 195 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, including Melanogenesis and Wnt signaling pathway etc. The results of current study revealed that the Yak coat color involved the regulation of various physiological process. These results provided helpful data to understanding on the molecular mechanisms of yak coat pigment.

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Marker assisted introgression of opaque-2 gene into elite maize inbred line BML-7

- Krishna M.S.R. and Sokka Reddy S.

Development of QPM (Quality Protein Maize) with high lysine and tryptophan is of foremost importance in maize breeding programme. Marker assisted selection in back cross breeding is the easiest way of developing QPM hybrids in less time. The present investigation was to attempt for conversion of elite maize inbred line BML-7 into QPM line. CML-181 was identified as a donor which showed good polymorphism with BML-7 for opaque-2 gene specific marker umc1066. Non-QPM inbred line BML-7 was crossed with QPM donor CML-181 and produced F1 followed by BC1F1 and BC2F1 population was developed. Foreground selection was carried out with umc1066 in F1 and selected plants were used for BC1F1 and BC2F1 populations. Two hundred plants were screened in both BC1F1 and BC2F1 population with umc1066 for foreground selection. The selected plants were screened for foreground selection with amino acid modifiers. Foreground selected plants for both opaque-2 and amino acid modifiers were screened for background selection for BML-7 genome. Recurrent parent genome (RPG) was calculated for BC2F1 population plants. Two plants have shown with RPG 90 – 93 % in two generation back cross population. The BC2F2 population derived from marker identified BC2F1 individuals were subjected to foreground selection followed by tryptophan estimation. The tryptophan and lysine concentration improved in all the plants. BC2F2 lines developed from the hard endosperm kernels were selfed for BC2F2 lines and the best line was selected to represent the QPM version of BML-7, with tryptophan concentration of 0.96% and lysine 4.01% in protein. QPM version of BML-7 line can be used for the development of QPM version maize single cross hybrids.

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Study of genetic variation among diverse barley (Hordeum vulgare L.) genotypes using agro-morphological traits and RAPD, ISSR, AFLP markers

Cheghamirza Kianoosh, Zarei Leila, Zebarjadi Ali Reza and Jalali Honarmand Saeid

The evaluation of different genetic similarity indicators can assist to produce better designs for future breeding programs. In this study, RAPD, ISSR and AFLP markers and agro-morphological traits were used to investigate the genetic relationships in some barley genotypes. Phenotypic data were collected under water-limited (rainfed) and irrigation conditions during 2006-2007 and 2007-2008 crop seasons. Polymorphism was observed in reactions with 31 out of 34 tested RAPD primers, 20 out of 24 tested ISSR primers and 7 primer combinations in AFLP technique. ISSR and RAPD fingerprinting detected more polymorphic loci (44%) than the AFLP technique (18%). The average polymorphic information content (PIC), resolving power (Rp), marker index (MI) and effective multiplex ratio (EMR) values derived from the three different marker systems differed indicating that they vary in their discriminatory power (ISSR > AFLP > RAPD). RAPD primers OPC07, UBC51, E10 and UBC28, ISSR primers UBC841 and UBC864 and AFLP primer M-CAA + E-AAC were significantly more informative primers to distinguish barley genotypes. Out of three molecular marker systems, only RAPD-based genetic distance showed a poor correlation with the agro-morphological based distance under rainfed conditions. Either of three molecular markers systems were not significantly correlated with phenotypic trais under irrigation conditions.

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PCR-RFLP screening of cry gene profiles in Bacillus thuringiensis and characterization of a novel cry2Ab8 gene

Huang Tianpei and Guan Xiong

The cry genes of Bacillus thuringiensis (Bt) encode economically significant insecticidal endotoxins. In the present study, the cry-type genes of 31 Bt isolates (six standard strains and 25 Wuyi Mountain isolates) have been analyzed by PCR-RFLP-based technique. Bt isolates harboring cry1 represented 87.1% of the isolates, while 67.7% and 3.2% of the isolates possessed cry2 and cry11 genes respectively. None of the isolates harbored cry3 to cry10 genes. The cloning and sequencing of a cry2Ab-type gene, designated as cry2Ab8, from the Wuyi Mountain isolate BRC-HZP2 revealed an open reading frame (ORF) of 1902 bp, encoding a protein of 633 amino acid residues. Both nucleotide and amino acid sequences similarity analyses showed that cry2Ab8 was a novel cry2Ab-type gene, with some major difference from the other cry2-type genes. So, the gene may service as a useful gene source for genetic manipulation leading to strain improvement of biopesticides or constructing new transgenic plants with resistance to insect pests.

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Quality evaluation and storage studies of legume based wadis (traditional delicacies) formulated using Sechium edule – An underutilized vegetable crop of North East India

Mishra K. Lokesh, Das Puspita and Gangmei Angela

The present research was carried out to formulate wadis from black gram paste and Sikut (an underutilized vegetable of North East India). The products were analyzed to determine their physicochemical attributes, bioactive components, organoleptic and shelf life quality. The study used Sikut (blanched and mashed vegetable) mixed with black gram in varying ratios. Sensory studies indicated that wadis with Sikut (40%) and black gram (60%) were most acceptable. These were subjected to physico-chemical, phytochemical, organoleptic evaluation and shelf life studies. The wadis supplemented with blanched and mashed Sikut vegetable exhibited higher ascorbic acid and total polyphenol content. Studies indicated that wadis formulated by substituting vegetable had better physico-chemical attributes except protein content as compared to wadis formulated with black gram (control). Significant differences were observed in the sensory quality of the products prepared from blanched vegetable and the control wadis. Storage studies indicated that the Sikut wadis can be stored for 12 months without any significant loss in its nutrient components and has no spoilage due to bacteria or yeast/moulds.

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Development of a synthetic medium for the growth of Catenaria anguillulae Sorokin: a facultative endoparasite of nematodes

Vaish Shyam Saran

Catenaria anguillulae is a facultative endoparasite of nematodes. This fungus seems to have very complex nutritional requirement. In view of this attempts were made to refine a medium suitable for this fungus. Out of the 24 media initially prepared for the growth of C. anguillulae, only medium no. 18 initiated the growth of the fungus. This medium was modified by varying concentrations of minerals of which medium no. 18C supported some growth of the fungus. Further on modification of medium no. 18C with various combinations of amino acids along with choline, when tested for growth of C. anguillulae, the fungus showed better growth on medium no. 18C3. Different sources of carbon were tested using medium no. 18C3. The maximum growth of the fungus was obtained on sucrose followed by dextrose. However, colony of the fungus on dextrose was well developed with better fungal mat than that of sucrose. Again, the above medium was standardized with different levels of dextrose which indicated that dextrose at the rate of 5g-l was best for supporting growth of C. anguillulae. Certainly, the medium defined relatively simple and closure to a complete synthetic medium. Further, above studies on nutritional requirement provide a good base for the selection of substrates for its mass culture.

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Enhancement of Skin Anti-Wrinkling Effects of Stichopus japonicus by Lactic Acid Fermentation

Lee Hyeon Yong

In this work, it was proved that anti-skin wrinkling ability of Stichopus japonicus extract was improved through lactic acid fermentation process with Lactobacillus rhamnosus GG (FE), compared with those from conventional extraction processes such as hot water (WE) and 70% ethanol extract (EE). Specifically, the amount of canthaxanthin, one of major bioactive substances of S. japonicas in the FE was increased up to 297.2 mg 100g-1 compared with 39.5 mg 100g-1 in the WE and 232.2 mg 100g-1 in the EE respectively. The fermentation extract also showed the highest DPPH scavenging activity at 22.48% at a concentration of 1.0 mg ml-1. For skin, anti-wrinkling effects of the extracts, the fermentation extract showed elastase inhibition ratio of 24.09% and MMP-1 and collagen production of 1313.2 pg ml-1 and 51.6 ng ml-1 from skin fibroblast cells respectively whose values are at least 20-45 % higher than those from the WE and EE. These results strongly indicate that the extraction associated with the fermentation process at relatively low temperature could minimize the destruction of the bioactive component, thereby increasing the contents of bioactive component of sea cucumber. This enhancement could also be caused by synergistic effects with other biologically active components such as Lipoteichoic Acid (LTA) in lactic acid bacteria.

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Structural and Photoluminescence Characteristics of Ultra Small Size Gold Nanoparticles Synthesized on ZnO Nano Substrate

Azimi Mina, Seyed Sadjadi Mirabdullah, Farhadyar Nazanin and Karamipour Shahnaz

In this work, we report synthesis and characterization of ultra-small size gold nanoparticles by wet chemical precipitation method using ZnO as a support. Zn-O nanoparticles were firstly prepared and gold coated then by reducing gold chloride solution. Structural and elemental characterization of the samples were performed using X-ray powder diffraction (XRD) patterns, Fourier transform infrared spectroscopy (FT-IR), Transmission electron and field emission scanning electron microscopy (TEM and FESEM). Elemental and spectral analysis of the samples, carried out using energy-dispersive X-rays (EDX) analysis and photoluminescence emission spectra revealed formation of ultra-small gold nanoparticles on ZnO nano-substrate with strong metal–substrate interactions (SMSI) between the two nanocomponents. Such a nanocomposite structure with enhanced photoluminescence spectra under visible light excitation shows that this benign biocompatible nanostructure can easily provide effective contact between biological tissue – nanoparticles required in biological detection, photocatalysis and dye-sensitized solar cells.

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Recent Advances in Anthocyanin Biosynthesis in Colored Wheat

Diddugodage Chamila Jeewani and Wang Zhong Hua

Anthocyanins belong to the flavonoid group of polyphenolic compounds which are responsible for the various colors including red, purple and blue color of different plant parts. Many studies have proven the health-promoting attributes of anthocyanins. These compounds are therefore considered to be a functional food factor which may have important implications in the prevention of chronic diseases. Anthocyanin in colored wheat is a very interesting topic as it can easily fulfill the daily anthocyanin requirement for a healthy life. Majority of the regulatory and structural genes for anthocyanin biosynthesis in different colored wheat have been identified and mapped in to several chromosomes. The expression of purple color in wheat has been extensively studied in different parts of wheat plant and the molecular genetic mechanism and regulation of anthocyanin biosynthesis has been discovered. However, there is still a gap for blue wheat for identifying their structural and regulatory genes. Blue and purple wheat can be identified as the potential functional food which can easily reached to the whole world. The facts about the genetic basis of wheat anthocyanin biosynthesis will be providing a valuable source to create nutritionally superior wheat varieties and further crop improvement programs.

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An efficient screening and identification technology for transgenic rubber tree (Hevea brasiliensis Muell. Arg)

Huijun Zhang and Yangrui Han

The method of this study is to construct GFP fusion expression vector using Agrobacterium-mediated leaf discs transformation to transform rubbers so as to obtain Transgenic Plants; green fluorescent protein (GFP) is used as a reporter gene and transgenic callus is observed under fluorescence microscope to verify whether the foreign gene was successfully transformed into Hevea brasiliensis.

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Anti-bacterial, Anti-Fungal and Cytotoxic Activities of Hydrazones and their Metal Complexes: A Review

Suvarapu Lakshmi Narayana and Baek Sung-Ok

Hydrazones and/or their metal complexes are well known to the biologist due to their biological properties such as anti-bacterial, anti-fungal, cytotoxic and anti-oxidant activities. Most of the newly synthesized hydrazones were tested for their different biological activities. Sometimes, the individual hydrazones are proved as better anti-bacterial, anti-fungal and cytotoxic agents than their metal complexes. In this review, we described the anti-bacterial anti-fungal and cytotoxic activities of hydrazones and their metal chelates reported since 2013. From this review, it is clear that still there is a gap of knowledge between the anti-bacterial, anti-fungal and cytotoxic activities of hydrazones and to their metal complexes.

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Physiological function of rhamnogalacturonan lyase genes based in the analysis of cis-acting elements located in the promoter region

Berumen-Varela Guillermo, Rivera-Domínguez Marisela, Troncoso-Rojas Rosalba, Báez-Sañudo Reginaldo and Tiznado-Hernández Martín-Ernesto

Rhamnogalacturonan lyase (RGL) recognizes and cleaves the α-1,4 glycosidic bonds located between L-rhamnose and D-galacturonic acids in the main chain of rhamnogalacturonan-I, one of the polysaccharide of the plant cell wall pectin domain. Although the biochemical mechanism of RGL is well known, its physiological role is not well understood. In this work, the published experimental data was reviewed analyzing the function of several cis-acting elements present in the promoter sequences of 13 RGL from tomato (Solanum lycopersicum L.) and 8 RGL from Arabidopsis thaliana in order to draw conclusions about the functions of RGL enzymes in the plant physiology. The most frequently transcription factors binding sites present in A. thaliana and S. lycopersicum L. sequences were a group of homebox zip proteins (AG, ATHB-1, ATHB-5, ATHB-9) that are involved in cell expansion and the development of specific organs. Furthermore, the presence of other motifs suggested that rhamnogalacturonan lyase enzyme plays a role in several physiological functions in plants including growth, development, fruit postharvest ripening, sexual reproduction and abiotic stress response. The findings of the present work will help in the design of future studies with the objective to elucidate the RGL gene physiological function in plants.

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