International Journal of Biochemistry and Biotechnology

International Journal of Biochemistry and Biotechnology ISSN: 2169-3048 Vol. 5 (2), pp. 660-664, February, 2016.  © International Scholars Journals 

Full Length Research Paper

Suitability of growing the wood ear mushroom A. polytricha in Tamil Nadu by low cost substrates and to assess its yield potential at different locations

Kalam Modi Sachin and Abdul V. Tendulkar

Centre for Plant Protection Studies, Department of Plant Pathology, Tamil Nadu Agricultural University, Coimbatore, Tamil Nadu, India.

E-mail: [email protected]

Accepted 15 January, 2016

Abstract

Surveys were conducted in the hills of Nilgiris, Shervoys and Lower Pulneys during rainy season. A wood ear mushroom was collected from coffee plantations of Horticultural Research Station, Yercaud. The fungus was identified as Auricularia polytricha (Mont.) Sacc. based on cultural and morphological characters. Molecular characterization studies on A. polytricha with P. platypus, P. florida and P. eous, using ITS primers exhibited similarity in ITS lengths among the mushrooms. On gel electrophoresis, all the mushrooms amplified a fragment between 600-700 bp. The studies conducted at Mushroom Research and Training Centre, TNAU, Coimbatore revealed that the paddy straw+wheat bran (3:1) ratio recorded minimum days for spawn run (21.3 days), pin head formation (31.3) and first harvest (35.6 days). The same combination also recorded the highest yield of 147.6 g/bed bioefficiency of 59.04%. The total cropping period was also the minimum in the same treatment. In the trials conducted at Vijaya Mushrooms, Coimbatore (North), paddy straw+wheat bran (3:1) ratio again recorded a significantly higher yield of 132.0 g/bed and bio efficiency of 58.20% with minimum cropping period of 47.3 days. The yield performance trials conducted at Maha Mushroom, Kovaipudur, and Coimbatore (South) also revealed the same trend as paddy straw+wheat bran (3:1) again recorded significantly higher yield of 130 g/bed and bioefficiency of 52.00%.

Key words: Auricularia polytricha, substrates, spawn production, ITS primers.

International Journal of Biochemistry and Biotechnology ISSN: 2169-3048 Vol. 5 (2), pp. 654-659, February, 2016. © International Scholars Journals 

Full Length Research Paper

PCR technique to identify the transgenes Cry1Ac and Cry2Ab in clonned recombinant cells

Abdul V. Tendulkar and Sachin Kalam Modi

Department of Biosciences, Veer Narmad South Gujarat University, Surat-395007, Gujarat, India.

E-mail: [email protected].

Accepted 22 January, 2016

Abstract

Cloning is the process of producing populations of genetically-identical individuals that occur in nature when organisms such as bacteria, insects or plants reproduce asexually. Modern cloning vectors include selectable markers (most frequently antibiotic resistance markers) that allow only cells in which the vector, but not necessarily the insert, has been transformed to grow. Additionally, the cloning vectors may contain color selection markers which provide blue/white screening on X-gal medium. Nevertheless, these selection steps do not absolutely guarantee that the DNA insert is present in the cells. Further investigation of the resulting colonies is required to confirm that cloning was successful. This may be accomplished by means of PCR. TOPOTA cloning provides one step cloning strategy for direct insertion of Taq polymerase amplified PCR products into a plasmid vector ligation (TOPO Cloning Reaction). In the presence of salt in TOPO Cloning reaction by mixing PCR product and TOPO vector, for transforming bacterial strain into competent cells, DH51α is supplied with the kit. Analysing transformants take 10 white colonies and culture them overnight in LB medium containing 50 µg/ml ampicillin or 50 µg/ml kanamycin. The plasmids were analyzed by PCR to confirm the presence of insert, after which PCR product of the Cry1A(c) and Cry2Ab transgene of Bt-cotton was produced to be cloned and ligated with vector.

Key words: TOPO-TA Cloning, PCR, BT-Cotton, Recombinant and Non-Recombinant colonies.

International Journal of Biochemistry and Biotechnology ISSN: 2169-3048 Vol. 5 (2), pp. 647-653, February, 2016. © International Scholars Journals 

Full Length Research Paper

Induction of polyploidization in A. malaccensis and investigation of phytochemical constituents using Headspace-Solid Phase Microextraction

Jaclyn Yuna Kuok^1,3*, Bernard Wei Ibrahim², Awal Lee Chong⁴, Ziana B. Ashaari¹, Hannah O. Krishnan³, James Amber Peter⁵, Yasmin S. E.¹, and Nicholas Tan¹

¹Forest Research Institute Malaysia (FRIM), 52109 Kepong, Selangor, Malaysia.

²Agro-Biotechnology Institute (ABI), Jalan Eksotika, Ibu Pejabat MARDI, 43400, Serdang, Selangor, Malaysia

³Faculty of Biotechnology and Biomolecular Science, Universiti Putra Malaysia, 43400, Serdang, Selangor, Malaysia.

⁴Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600, Bangi, Selangor, Malaysia.

⁵Malaysian Biotechnology Corporation Sdn. Bhd., Level 23, Menara Atlan, 161B Jalan Ampang, 50450, Kuala Lumpur, Malaysia.

E-mail: [email protected]

Accepted 5 January, 2016

Abstract

Aquilaria malaccensis is an endangered agarwood-producing plant found in most Asian countries and has a very long history of use in traditional medicine as incense, aromatic and therapeutic oil. Polyploidization is an established method to enhance the desired horticultural characteristics in many ornamental plants. In some medicinal plants, it proves to have positive effects on the production of secondary metabolites and/or bioactive compounds. Thus, the aims of this study are to induce polyploidization in A. malaccensis and investigate the phytochemical constituents using Headspace-Solid Phase Microextraction (HS-SPME), coupled with Gas Chromatography-Mass Spectrometry (GCMS) analyses. The present study showed that the highest percentage of polyploidization in A. malaccensis samples was at 120 h of exposure time using 1 mM colchicine and 0.1 mM trifluralin on shoot tip and nodal segment, respectively. Through HS-SPME/GCMS, the phytochemical constituents in polyploid doubled the amount than in diploid samples. The phytochemical constituents increased 4-folds in stem and twice in roots of polyploid samples as compared to diploid. The seedling however, had higher number of phytochemical constituents due to age differences against the in vitro samples. Enhancement of phytochemical production through polyploidization in A. malaccensis will provide new breeds for commercial plantation program and support the effort to reduce over-collection of the species in the wild.

Key words: Aquilaria malaccensis, diploid, polyploid, colchicine, trifluralin, Head Space-Solid Phase Microextraction (HS-SPME), Gas Chromatography-Mass Spectrometry (GC-MS).

International Journal of Biochemistry and Biotechnology ISSN: 2169-3048 Vol. 5 (2), pp. 642-646, February, 2016. © International Scholars Journals

Full Length Research Paper

Phytochemical screening of solvent extracts from Callilepis laureola plant

Polokuma E. Anderson*, Kojibola Winke and Ereapuaduoh B. V

Medical Research Council-Indigenous Knowledge System [Health] Lead Program Unit,P.O. Box 19070, Tygerberg 7505, Francie van Ziyl Drive,Parow Valley, Cape Town, South Africa.

E-mail: [email protected]

Accepted 2 January, 2016

Abstract

Phytochemical screening of solvent extracts from Callilepis laureola plant was studied. Methanol and water extracts, respectively, revealed that phytochemical constituents found in both extracts were phenolics and gums. Reducing sugars were only observed in water extract while phytosterols and flavanoids were only found in methanol extract. Other phytochemical constituents such as saponins, glycosides, anthraquinone, proteins and amino acids were not found in any of the extracts. These phytochemical constituents were detected by colour changes depending on the analysis. Total ash content of the powdered material was found to be 6.90% w/w, water soluble ash was 69.67% w/w and sulphate soluble ash was 16.80% w/w. Thin layer chromatography (TLC) of the plant extracts was also observed and it was concluded that in each extracts, suitable mobile phase must be determined.

Key words: Callilepis laureola, phytochemical screening, solvent extracts, ash value, TLC.

International Journal of Biochemistry and Biotechnology ISSN: 2169-3048 Vol. 5 (1), pp. 637-641, January, 2016. © International Scholars Journals

Full Length Research Paper

An ethnobotany survey of medicinal plants to determine its usefulness in industry and biotechnology

*José Luan, Augusto Xavier and Pereira Mário Sérgio

Departament of Botany, Museu Nacional/UFRJ, Laboratório de Etnobotânica e Taxonomia de Traqueófitos, Quinta da Boa Vista s/n, CEP 20940-040, São Cristóvão, Rio de Janeiro, RJ, Brazil.

E-mail: josé[email protected]

Accepted 17 October , 2015

Abstract

An ethnobotanical survey was done in Quissmã City, situated north of Rio de Janeiro State, during the period of 2001/2002.The study was carried out to collect information of species of plants used in traditional medicine by local population, and compared this information with those found in European Patent Office (EPO) and Japan Patent Information Organization (JAPIO) in October, 2002.A total of 94 species belonging to 49 botanical families were listed. The most representative families were Myrtaceae (8),Asteraceae (7), Lamiaceae (5) and Solanaceae (5). A total of 91 patent documents on 18 species were described. From these results, it is considered necessary to intensify efforts towards investments in programs designed to ensure proper search, inventory, conservation and management of biodiversity, as well as a system for managing the related patent documents. It is extremely important to form multidisciplinary teams for researches involving patents and plant biodiversity.

Key words: Ethnobotany, medicinal plants, patents, intellectual property.

International Journal of Biochemistry and Biotechnology ISSN: 2169-3048 Vol. 5 (1), pp. 626-636, January, 2016. © International Scholars Journals

Full Length Research Paper 

Analysis of differentially expressed proteins in the exfoliated cells of normal and squamous cell carcinoma of the uterine cervix to define candidate markers for cervical cancer

VN Saritha¹, Jestin K George¹, Abdul Jaleel⁵, Arun Surendran⁵, Meenakshi Saravana kumar⁵, MC Kalavathy ²,  Thara Somanathan³,  P Rema⁴,   K Sujathan¹.

Divisions of Cancer Research¹, Epidemiology², Pathology³, surgical oncology⁴,  Regional Cancer Centre, Medical College Campus and  Proteomics Facility, Rajiv Gandhi Centre for Biotechnology⁵,Trivandrum, Kerala, India.

Corresponding author. E-mail- [email protected]; [email protected]. Tel. 9104712522282; Fax.91-0471-2447454 

Accepted 29 January, 2016

Abstract

Cervical cancer continues to take extra ordinary toll on the lives of women in India, which has been attributed to the lack of systematically organized population screening programmes. Cost effective screening tools are yet to be identified for low resource countries. Proteomic studies constitute a reliable way to determine the biological alterations induced by HPV infection. Aim of the study was to identify the differentially expressed proteins in the exfoliated cells of Squamous Cell Carcinoma. Protein extracted from exfoliated squamous cells of normal smears and squamous carcinoma cells was analyzed by using Liquid Chromatography-Mass spectrometry method and the DNA was subjected for multiplex PCR for HPV status. The malignant samples were positive for oncogenic HPV. Proteome profiling revealed unique expression of 104 proteins in SCC while 20 proteins were found up-regulated and 64 proteins were down regulated. Up-regulated proteins include different isoforms of cytokeratin, ras related proteins, annexin, endoplasmin, vimentin, gelsolin, actin and heats hock proteins etc which can be used to define novel markers of cervical intra epithelial neoplasia.

Key words: Cervical cancer, proteomics, squamous cell carcinoma, human papilloma virus, liquid chromatography mass spectrometry.