Research Journal of Biotechnology

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Genetic diversity of Toona sinensis in Korea and the phylogenetic relationship of this species based on chloroplast DNA and ribosomal DNA sequences

Sun Yan-Lin, Park Wan-Geun, Lee Hak-Bong, Kim Nam-Young, Choi Min-Hee and Hong Soon-Kwan

Meliaceae is a widely distributed subtropical and tropical angiosperm family. This family included approximately 50 genera encompassing 700 species, occurring in a variety of habitats, from rain forests and mangrove swamps to semi-deserts. As human actions increase and the climate changes, the growing environment of plants is changing and habitat destruction is threatening to erode the genetic resources of these species. Thus, the conservation of the genetic biodiversity has been widely recognized to be necessary. Cedrela and Toona are closely related genera. Although they have been indisputably considered independent genera, their genetic diversity and phylogenetics are still confused. In this study, the genetic diversity and phylogenetics of 17 Toona sinensis populations were determined based on the DNA sequences of regions of the chloroplast DNA (cpDNA) genes matK and rbcL and regions of the ribosomal DNA (rDNA) 5S gene and internal transcribed spacer (ITS) regions. The rbcL region exhibited little genetic variation. The 5S region showed the highest level of genetic variation, however, phylogenetic trees based on this region bore no obvious relationship to the geographic origins. The monophyly of Cedrela and Toona was strongly supported by analyses of the matK regions whereas the ITS results only supported the monophyly of Cedrela. This conclusion was supported by representative variable sites within the ITS regions, which were identical in T. sinensis and Cedrela but differed from the other Toona species. This study contributed better molecular discrimination between Cedrela and Toona and among Toona species.

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Does cytogenetics have answers to genomic instability in cancers with Leukemia as the most studied example?

Jamil Kaiser, Reddy H., Natukula K. and Jayaraman A.

Cancer cells usually exhibit abnormal karyotypes, leukemia is generally diagnosed based on karyotyping results and thus a number of questions which can arise are: can these abnormal chromosomes cause cancer or are they a consequence of tumorigenesis? Can a single gene mutation drive the neoplastic transformation? These genomic changes present researchers with a chicken-and-egg dilemma: the question is- does this genomic instability contribute to cancer? or is it simply a byproduct of cellular processes operating out of control? One assumption is clear and proven in the very beginning of tumorigenesis that cytogenetic alterations are directly linked to tumor formation, progression and/or metastases. In this study we present our data on leukemia diagnosis as an example to discuss in length the importance of cytogenetics in prognosis as this has been a neglected field in early cancer detection. It is seen that genes affected by chromosomal rearrangements like transversions, mutations, trisomies, aneploidy or fusion genes like Philadelphia chromosomes are involved in critical stages in cell division, may lead to cancer development. Thousands of chromosomal aberrations have been discovered in different types of cancer. But how do these various changes all hijack normal cellular processes to promote cancer?. Aneuploidy, or an incorrect number of chromosomes, is often seen in cancer cells, but its cause and effects have been unclear. A few predictions include that a mutated "caretaker" gene, which usually stabilizes cellular DNA, plays a key role in the genetic instability that is the hallmark of many forms of cancer.

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Carbon and nitrogen in leaves, branch and stem of 19 different Mediterranean maquis species at the same site, Isparta-Turkey

Karatepe Yasin

The maquis is widely distributed across areas in Turkey where the Mediterranean climate predominates as well as the Mediterranean basin and Europe. Maquis vegetation, which harbors many different species of plants, spreads horizontally and vertically over large areas mainly in the Mediterranean Region of Turkey and forms important ecosystem within Turkey. This study investigated within the same habitat (Isparta-Turkey) shows the carbon (C), nitrogen (N) and C/N ratios in the leaves, branches and stems of 19 different plant species (1-Laurus nobilis L., 2-Tamarix parviflora DC., 3-Cercis siliquastrum L., 4-Olea europaea L., 5-Rhus coriaria L., 6-Paliurus spina-christi Mill., 7-Vitex agnus-castus L., 8-Phillyrea latifolia L., 9-Pistacia terebinthus L., 10-Quercus coccifera L., 11-Fontanesia phillyreoides L., 12-Cistus creticus L., 13-Nerium oleander L., 14-Myrtus communis L., 15-Arbutus andrachne L., 16-Crataegus monogyna Jacq., 17-Styrax officinalis L., 18-Cotinus coggygria Scop. and 19-Juniperus oxycedrus L.) with significant distribution in the maquis shrublands of the Mediterranean Region of Turkey. In general, the carbon and nitrogen ratios of different plant species demonstrated significant differences.

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A dose dependent cytogenetical disturbances induced by 2, 4-dichlorophenoxy acetic acid and isoproturon in root tip cells of Triticum aestivum L.

Kumar Sanjay

Concentration range (0-250 ppm) of two herbicides 2, 4-Dichlorophenoxy acetic acid and Isoproturon was applied on the wheat (Triticum aestivum L.) seedlings for 72h at normal laboratory light/dark condition and temperature (25±20C) during winter (October-December, 2010). The root tips length (1-2 cm in size) of treated and control were collected and fixed in 3:1 ethanol: acetic acid for 24h followed by preservation in 70% alcohol for further observations. The slides were prepared in 2% acetocarmine to observe the abnormalities induced by the tested herbicides. It has been observed that stickiness and bridges were the most common among the chromosomal abnormalities at all the concentrations used. Besides the stickiness and bridges formation in the chromosomes, other abnormalities like chromosomal fragments, star-shaped structure of chromosomes, multipolarity in chromosomes, dysjunctions of chromosomes, overcontractions, splits among chromosomes, unequal distribution and un-oriented chromosomes were observed. It is obvious from the above abnormalities that both the herbicides used in agricultural practice are toxic to mitotic activity for the cause that of the induced cytological disturbances in root tip cells of wheat (Triticum aestivum L.).

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Genetic diversity of mulberry (Morus spp.) genotypes using RAPD markers

Wani Shabir A., Bhat M. Ashraf, Kamili Afifa. S., Wani Nawsheeba, Akhter Sabeena and Majid Naina

India is recognized as one of the twelve mega biodiversity centers of the world and harbors 11.9% of the world flora. The plant genetic diversity is being recognized by the International Conservation Community as a basic key component for the sustainable development of agriculture, horticulture and sericulture. Rich source of wild relatives of mulberry flora are reported to occur in Himalayan belts. Germplasm evaluation is essential in any crop improvement programme and genetic characterization at morphological and molecular level is very vital for breeding programs to be successful. Thirty RAPD markers generated a total of 179 alleles across 17 genotypes and the number of alleles per locus varied from 4 (OPA 13) to 9 (OPBB 13) with an average of 5.96 per locus. The similarity coefficient ranged from 0.74 to 0.93 with an average of 0.84. The highest similarity value 0.93 was recorded between SKM- 33 and KNG and SKM -33 and Goshoerami, lowest similarity coefficient (0.74) was found between local mulberry and Kokuso- 20. Cluster analysis by Unweighted Pair Group Method using Arithmetic average (UPGMA) grouped 17 mulberry genotypes/selections into 8 different clusters. Cluster-2 contain highest (6) genotypes.

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Genetic diversity analysis of maize verities based on SSR markers

Changsheng Li, Muhammad Irfan, Chunyu Zhang and Feng Lin

In the present study sixty seven maize verities were used to asses the genetic relationship among them using eleven PCR based simple sequence repeats (SSR) marker. The observed number of alleles and effective number of alleles was found to be average of 3.0 and 2.45 respectively. The expected homozygosity, heterozygosity and Nei’s heterzygosity values were average of 0.435, 0.5685 and 0.5643 revealing heterotic population respectively. Shannon’s diversity index and PIC value produced by all primers were 0.9391 and 0.488. A dendrogram was constructed based on UPGMA method which clustered into three main groups based on their genetic diversity which was further confirmed by the principal coordinate analysis. These results demonstrated that SSR markers successfully detected the genetic diversity in maize which could be further very helpful in improving the maize yield.

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Structural elucidation of a chemical compound from mollusk, Drupa margariticola against fish pathogens

Chellaram C.

The mollusk, Drupamargariticola were collected from the Tuticorin coastal waters of Southeastern India.The crude extract showed promising antibacterial activity against some selected fish pathogens. The crude extract of D. margariticola was purified using chromatography (HPTLC and HPLC) and characterized by spectroscopy (NMR, Mass and IR) techniques. The pure compound, diterpenoidacylglycerol, [(CH3)5, (CH2)7, (CH)4, (CH-OH), (C=CH), (CHO), (CH2-O) with ester or ether functionalities] possessed potential antibacterial activity against Serratiamarcescens, Proteusmirabilis, Aeromonashydrophila and Vibrio harveyipathogens. The encouraging biological activities obtained in this study suggest that the tropical marine environment is a potential source for new bioactive compounds.

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Molecular Cloning and Sequences Analysis of SS Gene from Panax japonicus

Lai Zhang and Min Sun

The squalene synthase (SS) from Panax japonicus is an important regulatory enzyme in the biosynthetic pathway of triterpenoid saponins (TS). Regulating the SS activity directly affects the synthesis of squalene (SQ), as well as the synthesis of TS for which SQ acts as the precursor. By using the homologous cloning program RACE, the SS coding for the key enzyme gene in the synthetic TS pathway of P. japonicus was successfully cloned. It was named PjSS. Its fragment was 1353 bp, encoding 415 amino acids. The PjSS protein was an unstable and hydrophobic protein, with molecular mass of 109559.7 kD. P. japonicus SS was located in the chloroplast transit peptide (cTP), mitochondria transit peptide (mTP) and secretory pathway (SP). Its first transmembrane domain was from A281 to N303 and the second was from S385 to S407. Eight structural motif sites were present. The secondary structure of PjSS was composed of α-helix, random coil, extended strand and β-angle of 67.47%, 22.41%, 7.23% and 2.89% respectively. The protein exhibited folding properties to form the typical three-dimensional SS structure.

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In vivo evaluation of Acute and Chronic Anti-Inflammatory activity of Ethanol leaf Extract of Vitex Negundo Linn.

Murugesan Deepa, Ponnusamy Renuka Devi and Gopalan Dhanalakshmi Karur

The ethanol leaf extract of Vitex negundo Linn was investigated for anti-inflammatory activity at the doses (p.o.) of 200 mg/kg and 400 mg/kg body weight. For evaluation of inflammation carrageenan induced paw edema served as acute model and cotton pallet induced granuloma served as a chronic model in Albino Wister rats. The higher doses of VLE (200 and 400 mg/kg, p.o.) were inhibiting carrageenan induced paw edema successfully. In cotton pallet granuloma inflammation was induced by implanting the sterilized pre weighed cotton pallet (10mg) subcutaneous in the ventral region of the groin. Treatment continued up to 8 days. The decrease in weight of the cotton pellet as compared to the control was considered as anti-inflammatory effect of Vitex negundo ethanol leaf extract. In both the inflammatory models ethanol leaf extracts showed more potency in terms of percentage of inhibition of inflammation. Both inflammatory model results revealed that the ethanol leaf extract of Vitex negundo had anti-inflammatory activity. From acute oral toxicity studies of Organization for Economic Co-operation and Department (OECD-423 guidelines), no mortality was observed even at highest dose of Vitex negundo ethanol leaf extract (VLE) 2000 mg/kg, p.o. GCMS was performed for the ethanol leaf extract of Vitex negundo. Various phenolic phytocompounds were identified through GCMS.

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Production of L-malate from glucose with Rhizopus oryzae

Ya Liu, Ying Yang, Ting Sun and Xingjiang Li

In order to improve the yield of L- malic acid, the fermentation process and glucose metabolism pathways are studied in this paper by optimizing the culture medium and fermentation conditions by using single factor experiment and response surface method (Box-Behnken).The results showed that the optimal medium compositions for malate production were as follows: glucose, (NH4)2SO4, MgSO4, FeSO4•7H2O, KH2PO4, ZnSO4, CaCO3 were 100 g/L, 4.0 g/L, 0.3 g/L, 0.025 g/L, 0.5 g/L, 0.1 g/L and 80 g/L respectively. The fermentation conditions by means of single factor experiments were found to be as follows: fermentation equipment for stirred tank, a temperature of 32 oC, ventilation volume of 3.0 L/m and rotation rate of 500 rpm. Spore suspension was incubated for 48 hours. The fermentation period lasted as long as 144 hours leading to the L-malic acid yield of 57.71 g/L. Thus, by fermentation processing optimization, the yield of malate was raised effectively.

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Effect of solution pH on partial properties of photosystem II from spinach

Liu Jianguo

Our previous work has demonstrated the isolation of photosystem I (PSI) from spinach using ultrafiltration with a final purity of 84%. In that process, the major impurity (photosystem II, PSII) was preserved in the permeate after ultrafiltration. In order to develop a practical bioseparation process, the mechanism of the new membrane-based process should be extensively studied. In this work, the effect of solution pH on partial properties of PSII was examined. Following the isolation of PSII from spinach using n-dodecyl-beta-D-maltoside as detergent, the low-temperature fluorescence, UV-VIS absorption, activity, atomic force microscopy imaging and thermal stability were determined. Solution pH was found to have a significant effect on the activity and thermal stability of PSII. The results also demonstrated that the “molecular sieve effect” may play a central role in PSI isolation using ultrafiltration.

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Non-invasive Breast Cancer detected by Patch Antenna Design

Sakthisudhan K., Tamilselvan G. M. and Thangaraj P.

This paper proposed breast cancer detection through microwave tomography systems and it introduced novel design of antenna. The breast cancer detection examined on the scattering losses in between healthy and cancerous breast tissues with help of antenna design. It is an easy way to detect the breast cancer and it is faster than anechoic chamber‘s near filed in the data collection method. The breast cancer detects the microwave spectrum of 1 to 10 GHz frequency range. Finally, detection criteria concludes the between scattering losses which are making over healthy and cancer breast tissues. The design consists of ΨU shaped antenna mounted on the strips with FR4 dielectric constant of 4.4. It covers ΨU shaped notch microstrip patch with wide band of 6.8-7.6 GHz frequency of operation. This novel design is suitable for breast cancer detection. It consists of an asymmetric structure with the total geometric dimension of 50×40×1 mm3. The design provides reflection loss about -22 dB at 7 GHz. The dielectric phantom breast model is used for simulation and provides reflection loss about -22dB at 3.6 GHz and -16dB at 6.5GHz. These simulation results are analyzed with the results obtained from fabricated models. The fabricated patch antenna provides reflection loss about -35.86 dB at 6.4 GHz. The results obtained from fabricated models are compared with simulation results.

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Impact of Salt on Morphological Characters and Hormonal Regulation in three Capsicum Varieties

Maurya Vaibhav Kumar and Gothandam K. M.

Present experiment is an attempt to assess the effect of salt stress on morphological characters and endogenous hormones on three capsicum varieties. The capsicum varieties CO1(salt sensitive), K2(moderate salt tolerant) and G4(salt tolerant) were subjected to different concentration of NaCl (25mM, 50mM, 100mM, 150mM and 200mM) for 30 days in controlled green house condition. Result clearly shows the significant decrease in leaves size, fruit weight and plant height and chlorophyll content of all varieties. Decrease in zeatin, auxin and gibberellic acid was highly correlated to tissue damage in individual varieties but ABA expressed increases with lower salt concentration while decreases with high salt treatment for each variety. 200mM salt treatment was found most detrimental for CO1 variety leading to death on 35th day of treatment.

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Propamocarb effects on antioxidant parameters and osmolyte levels of cucumber fruit differing in propamocarb residual capacity

Peng Wu

The aim of this study was to investigate the effect of fungicide propamocarb (PM) on cucumber fruit differing in propamocarb residual capacity. In respect of antioxidant parameters, effect of PM application on peroxidase (POD) and glutathione S-tramsferase (GST) activities of low PM residual cultivar (D0351) was higher than control at 5, 15 and 25 day after treatment (DAT). With osmolyte levels, only soluble sugar content of D0351 following PM application significantly increased at 5, 15 and 25 DAT. However, high PM residual cultivar (D9320) was no significant change with control at 5, 15 and 25 DAT. The physiological and biochemical parameters measured indicated that PM application affected the activities of POD and GST and the content of soluble sugar only in low PM residual cultivar (D0351). This may imply that the D0351 plant had the capacity to elicit differential responses to PM application in order to reduce the systemic magnitude of stress.

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