Indian Journal of Biotechnology

 

http://www. niscair.res.in

Total visitors: 4,073  since 17-04-09

 

VOLUME8

CODEN: IJBNAR 8(2) (2009) 141-248

NUMBER2

APRIL2009

ISSN: 0972-5849

 

  CONTENTS

 

 

Reviews

 

Seed storage protein gene regulation—A jig-saw puzzle

147

        Rajesh Mehrotra, Sanjeev Kumar, Sandhya Mehrotra & B D Singh

 

 

 

Biofilm: Importance and applications

159

        C R Kokare, S Chakraborty, A N Khopade & K R Mahadik

 

 

 

Leptin: A biomolecule for enhancing livestock productivity

169

        R Agarwal, P K Rout & S K Singh

 

 

 

Towards modeling and design of vermicomposting systems: Mechanisms of composting/
vermicoposting and their implications

177

        Tasneem Abbasi, S Gajalakshmi & S A Abbasi

 

 

 

Papers

 

Modification of overlap extension PCR: A mutagenic approach

183

        Darshan H Patel, Seung Gon Wi & Hyeun Jong Bae

 

 

 

A simplified method for extraction of high quality genomic DNA from Jatropha curcas for genetic diversity and molecular marker studies

187

        D V N Sudheer Pamidimarri, Meenkashi, Ritam Sarkar, Girish Boricha & Muppala P Reddy

 

 

 

Prediction of T cell epitopes for the utility of vaccine development from structural proteins of dengue virus variants using in silico methods

193

        Pallavi Somvanshi & P K Seth

 

 

 

Development of Trichoderma harzianum endochitinase gene construct conferring antifungal activity in transgenic tobacco

199

        G V S Saiprasad, J B Mythili, Lalitha Anand, C Suneetha, H J Rashmi, C Naveena &
Girija Ganeshan

 

 

 

STS marker based tracking of slow rusting Lr34 gene in Indian wheat genotypes

207

        Priyamvada, Ratan Tiwari, M S Saharan, R Chatrath, Priyanka Siwach & B Mishra

 

 

 

Kappa-casein gene polymorphism in Indian goats

214

        A Kumar, P K Rout, A Mandal & R Roy

 

Acclimatization of neem microshoots adaptable to semi-sterile conditions

218

        M Lavanya, B Venkateshwarlu & B Poornasri Devi

 

 

 

Micropropagation of Baliospermum montanum (Willd.) Muell. Ag.—A threatened medicinal plant

223

        S Sasikumar, S Raveendar, A Premkumar, S Ignacimuthu & P Agastian

 

 

 

Efficacy of non-purine and purine cytokinins on shoots regeneration in vitro in sugarcane

227

        Vandana Vinayak, Ashok K Dhawan & V K Gupta

 

 

 

Rapid clonal multiplication through in vitro axillary shoot proliferation of Centella asiatica L.

232

        K Karthikeyan, C Chandran & S Kulothungan

 

 

 

Short Communications

 

In vitro propagation of Caralluma sarkariae Lavranos & Frandsen―An endemic and endangered medicinal plant

236

        V Raja Sreelatha, S Sandhya Rani, P V Krishna Reddy, M Naveen, A Ugraiah & T Pullaiah

 

 

 

Microflora (fungal and bacterial) of selected terrestrial and marshy species of rhizosphere in response to spent wash treatments

240

        P K Singh & K P Sharma

 

 

 

Instructions to Contributors

245

 

 

      

 

AUTHOR INDEX

 



 

 

 

 

Indian Journal of Biotechnology

Vol 8, April 2009, pp 147-158

 

 

Seed storage protein gene regulation—A jig-saw puzzle

 

Rajesh Mehrotra1, Sanjeev Kumar2, Sandhya Mehrotra3 and B D Singh*

 

School of Biotechnology, Faculty of Science, Banaras Hindu University, Varanasi 221 005, India

1Biological Science Group, Birla Institute of Technology, Pilani, 333 031, India

2Biotechnology Laboratory, Indian Institute of Vegetable Research, P O Jakhini-Shahanshahpur, Varanasi 221 305, India

3Laboratory of Plant Molecular Morphogenesis, Faculty of Biological Sciences, Nara Institute of Science and Technology,
8916-5, Takayama-cho, Ikoma-city, Nara Prefecture-630-0101, Japan

Received 23 April 2008; revised 22 September 2008; accepted 4 December 2008

Seed storage proteins are synthesized in high abundance during seed development and maturation, and are characterized as mid-embryogenesis and late-embryogenesis abundant proteins. The regulation of transcription of seed-protein gene families is not coordinated and transcription of each gene family is independently regulated. All seed protein gene families are regulated, in part, at the transcriptional level and a number of cis-acting elements and trans-acting factors are involved in this regulation. In many cases, a combinatorial interaction among transcription factors plays a critical role. In addition, CpG methylation has been found to be responsible for maintenance of the seed protein genes in an inactive state in tissues other than developing seeds. In some cases, a type of post-transcriptional control has been reported. In these cases, even when the transcription rates of seed protein and non-seed protein genes are similar, their mRNA levels vary upto 10000-fold. Formation of hairpin-loop structure in zein mRNA has been reported to serve as a kind of translational control. Proteins like Hsp70/Bip have been reported to play a critical role in the developing endosperm. This review highlights the regulation of seed storage protein genes at various levels with a view to understand the biological mechanisms regulating their synthesis.

Keywords: Seed storage protein, gene expression, cis-elements, promoter

 

 

Indian Journal of Biotechnology

Vol 8, April 2009, pp 159-168

 

 

Biofilm: Importance and applications

 

C R Kokare*, S Chakraborty, A N Khopade and K R Mahadik

Department of Pharmaceutical Biotechnology, Poona College of Pharmacy, Bharati Vidyapeeth University
Pune 411 038, India

Received 24 December 2007; revised 18 August 2008; accepted 2 November 2008

Biofilm is an assemblage of the microbial cells that is irreversibly associated with a surface and usually enclosed in a matrix of polysaccharide material. Biofilm is composed primarily of microbial cells and extracellular polymeric substance (EPS). Extracellular polymeric matrix plays various roles in structure and function of different biofilm communities. Adhesion to the surface provides considerable advantages such as protection against antimicrobial agents, acquisition of new genetic traits, and the nutrient availability and metabolic co-operability. Anthony van Leeuwenhoek, who discovered microbial attachment to his own tooth surface, is credited with the discovery of biofilm. The formation of biofilm takes place in three steps. Biofilm is responsible for chronic bacterial infection, infection on medical devices, deterioration of water quality and the contamination of food. This article provides an overview of the formation of biofilm, structure, role in microbial communities and its applications.

Keywords: Biofilm, polymeric substance, hydrodynamics, probes, pathogenesis

 

 

Indian Journal of Biotechnology

Vol 8, April 2009, pp 169-176

 

 

Leptin: A biomolecule for enhancing livestock productivity

R Agarwal, P K Rout* and S K Singh

Genetics and Breeding Division, Central Institute for Research on Goats, Makhdoom, Farah, Mathura 281 122, India

Received 9 January 2008; Revised 19 September 2008; Accepted 25 November 2008

Livestock play a significant role by providing food security as well as income resource for less poor people in our country. It is necessary to enhance and sustain the production efficiency of livestock species in the changing face of population scenario to feed a large growing population. Therefore, it is important to adopt different strategies to increase productive efficiency of livestock. Leptin is one of the most useful biomolecule to act as a marker for identifying high performing individuals leading to better adaptability and productivity. Leptin has a pleiotropic effect on regulating apetite, energy metabolism, growth, reproduction, body composition and immunity. It is mostly produced in the white adipose tissue and informs the central nervous system (CNS) about the total fat depot of the body. Leptin is also involved in regulation of nutritional status and reproductive function and also regulates fetal development in livestock. Plasma leptin level increases linearly with increases in body fat mass. Leptin reduces feed intake in rodents, ruminant and other livestock species and also plays a role in energy expenditure. The physiological properties support leptin as a strong candidate gene for evaluation of genetic polymorphisms, which has further been associated with growth, milk yield and other economic traits in different livestock. The present review deals with the leptin structure and its implications in regulation of growth, reproduction, immunity and also enhancing livestock productivity.

Keywords: Genetic polymorphism, leptin, livestock, production performance

 

 

Indian Journal of Biotechnology

Vol 8, April 2009, pp 177-182

 

 

Towards modeling and design of vermicomposting systems: Mechanisms of composting/vermicomposting and their implications

 

Tasneem Abbasi, S Gajalakshmi and S A Abbasi*

Center for Pollution Control and Energy Technology, Pondicherry University, Puducherry 605 014, India

Received 29 April 2008; revised 29 September 2008 ; accepted 2 December 2008

Several studies have been reported, and are continued to be done by scientists especially in Asia, in which earthworms are added to one or other substrate undergoing composting. The concerned authors call it ‘vermicomposting’. Other authors use the term ‘vermicomposting’ to denote processes in which earthworms are made to feed upon one or other substrate, to generate a useful product (vermicast). Present review has embarked on a series of efforts aimed at clearly defining the mechanism of vermicomposting process and to model it. This, in turn, is envisaged to be made the basis for developing rational criteria with which vermireactors are to be designed and operated in a manner that maximizes the process efficiency and minimizes the production cost. During the course of these efforts, authors have conducted a detailed analysis of the steps associated with composting and vermicomposting. Based on an analysis of the experiments done earlier by authors, as also on the work published by others, it is now reported that composting and vermicomposting are essentially different types of processes involving different bioagents, process conditions, reactor operation strategies, and process control parameters. Hence, to achieve optimal results, the two processes should be run in isolation, composting should always precede vermicomposting, and never in combination. It is also suggested that the term ‘vermicomposting’ should be used only to denote the process in which reactor systems are used to transform biodegradable substrates into vermicasts.

Keywords: Composting, vermicomposting, modeling, design

 

 

Indian Journal of Biotechnology

Vol 8, April 2009, pp 183-186

 

 

Modification of overlap extension PCR: A mutagenic approach

 

Darshan H Patel1, Seung Gon Wi1 and Hyeun Jong Bae1,2*

1Bio-energy Research Institute and 2Department of Forest Products and Technology, Chonnam National University,
Gwangju 500-757, South Korea

Received 1 April 2008; revised 29 September 2008; accepted 10 December 2008

In vitro site-directed repair or creation of a mutation is an invaluable technique in genetic and protein engineering. Several methods have appeared in literature but still require many modifications. We describe a rapid and efficient modified overlap extension PCR method for multiple uses in mutagenesis studies. The protocol is based on two rounds of PCR with the help of two sets of primers, two flanking and two internal mutagenic primers. Two fragments of DNA prepared in first round of PCR are then allowed themselves to anneal in the second stage of PCR using gradient annealing temperature without using flanking primers. This protocol has been used for correcting a mutation caused in exoglucanase (CBHII) gene of Trichoderma spp. We successfully synthesized the full length of gene from two fragments in the second round of PCR in lesser time.

Keywords: DNA polymerase, ex taq, overlap extension PCR, site directed mutagenesis, one step overlap extension-PCR

 

 

Indian Journal of Biotechnology

Vol 8, April 2009, pp 187-192

 

 

A simplified method for extraction of high quality genomic DNA from
Jatropha curcas for genetic diversity and molecular marker studies

D V N Sudheer Pamidimarri, Meenakshi, Ritam Sarkar, Girish Boricha and Muppala P Reddy*

Discipline of Wasteland Research, Central Salt and Marine Chemicals Research Institute, Bhavnagar, 364 002, India

Received 8 June 2007; revised 1 April 2008; accepted 20 August 2008

A simple and efficient protocol for the extraction of high quality genomic DNA from different tissues, including callus generated from leaves of Jatropha curcas has been developed. The important steps in this protocol include (a) use of 3.5 M NaCl in extraction buffer; (b) 2.0 M NaCl (final concentration) during precipitation; (c) Tris saturated phenol in place of phenol:chloroform:isoamyl alcohol at purification phase; (d) 80% ethanol for DNA precipitation, and (e) performing all the steps at RT. The DNA thus extracted from the leaves had 1.81±0.063, OD at A260/280 and the yield was 120 to 140 µg/g of material. The extracted DNA was found suitable for restriction digestion, ligation and PCR amplification. It was also used for DNA fingerprinting techniques, RAPD and AFLP, for development of molecular markers and studies on genetic diversity.

Keywords: CTAB, Jatropha curcas L, molecular markers, RAPD, AFLP

 

 

Indian Journal of Biotechnology

Vol 8, April 2009, pp 193-198

 

 

Prediction of T cell epitopes for the utility of vaccine development from structural proteins of dengue virus variants using in silico methods

 

Pallavi Somvanshi* and P K Seth

Bioinformatics Centre, Biotech Park, Sector-G, Jankipuram, Lucknow 226 021, India

Received 1 February 2008; revised 29 September 2008; accepted 5 December 2008

The spread of dengue virus, dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS) has increased significantly in the past two decades and thus has been a major concern of public health globally. Dengue virus infection can lead to a wide range of manifestations in the form of undifferentiated fever, classic dengue fever, DHF with plasma leakage, which leads to hypovelmic shock DSS. A new strategy for developing prophylactic and therapeutic application of pathogen-specific immunity was provided from epitope-based vaccines; it is a critical requirement for the identification and selection of T cell epitopes that act as vaccine target. Immunoinformatics serves as a valuable tool to screen and select antigenic peptide sequences as potential T cell epitopes for binding affinity with HLA alleles. We studied dengue variants conserved epitopes in three structural proteins, capsid, envelope and precursor membrane, which recognize some highest binding affinity HLA. A total of 45 promiscuous nanomer candidate epitopes for HTL are recognized against MHC Class II and 28 promiscuous epitopes are recognized against CTL for MHC class I. This computational prediction analysis will improve our understanding of T cell immune response and help in identifying the antigenic peptide for formulation of antigen based diagnostic kit and peptide based subunit vaccine design against dengue virus.

Keywords: Dengue, epitopes, HLA, vaccine, in silico, structural proteins

 

 

Indian Journal of Biotechnology

Vol 8, April 2009, pp 199-206

 

 

Development of Trichoderma harzianum endochitinase gene construct
conferring antifungal activity in transgenic tobacco

 

G V S Saiprasad1, J B Mythili1*, Lalitha Anand1, C Suneetha1, H J Rashmi1, C Naveena1 and Girija Ganeshan2

1Division of Biotechnology and 2Division of Plant Pathology, Indian Institute of Horticultural Research
Hessaraghatta, Bangalore 560 089, India

Received 26 November; revised 11 September 2008; accepted 20 November 2008

Trichoderma harzianum is a popular biocontrol agent used extensively against phytopathogenic fungi. The mode of action of this fungus is through secretion of cell wall degrading enzymes including chitinases. Thus, the chitinase genes isolated from T. harzianum have been successfully utilized in the production of transgenic plants with enhanced resistance to several fungi. The stringent rules of IPR necessitates that genes are cloned and constructs are developed from the local isolates of this fungus. In view of which full length chitinase gene was isolated and a construct was developed under the expression of constitutive promoter CaMV 35S for use in plant transformation. Expression of chitinase gene in transgenic tobacco plants was found to be higher as revealed by the endochitinase assay and relative quantitative RT-PCR. Its efficacy in inhibiting the fungal growth was also reported in vitro as well as in vivo in the detached leaves of tobacco transformants containing the gene. Thus, the development of T. harzianum chitinase gene construct (pIIHR-Th-Chit) would facilitate the exchange of the construct amongst researchers in the country for the development of fungus resistant transgenic plants.

Keywords: Chitinase, disease resistance, pathogen inhibition assay, transformation, Trichoderma harzianum

 

 

Indian Journal of Biotechnology

Vol 8, April 2009, pp 207-213

 

 

STS marker based tracking of slow rusting Lr34 gene in Indian wheat genotypes

Priyamvada1, Ratan Tiwari*1, M S Saharan1, R Chatrath1, Priyanka Siwach2 and B Mishra1

1Directorate of Wheat Research, Karnal 132 001, India

2Department of Bio and Nanotechnology, Guru Jambheshwar University, Hisar 125 004, India

Received 17 October 2007; revised 22 September 2008; accepted 28 November 2008

Bi-allelic STS marker was used to confirm the presence of adult plant durable rust resistance gene Lr34 in advance generation breeding lines. These lines were scored for leaf rust three times at an equal interval and the area under disease progress curve (AUDPC) was calculated. The lower AUDPC values of Lr34 positive lines confirmed their slow rusting nature. In the absence of direct selection method, the breeders are selecting Lr34 gene carrying lines unintentionally as they showed better resistance. Lines possessing Lr34, an ‘undefeated gene’, should be used in breeding programme in order to have a broad-spectrum durable leaf rust resistance.

Keywords: AUDPC, leaf rust, Lr34 gene, Puccinia triticina

 

 

Indian Journal of Biotechnology

Vol 8, April 2009, pp 214-217

 

 

Kappa-casein gene polymorphism in Indian goats

A Kumar, P K Rout*, A Mandal and R Roy

Division of Genetics and Breeding, Central Institute for Research on Goats (CIRG) Makhdoom, Farah (Mathura) 281 122, India

Received 9 January 2008; revised 19 September 2008; accepted 22 November 2008

The present work was carried out to analyse the k-casein variants in five Indian goat breeds by SDS-PAGE, PCR-RFLP and SSCP method. A total of 152 unrelated blood samples and 102 milk samples belonging to Barbari, Beetal, Marwari, Surti and Local MP (non-descript) goats were used for analysis. SDS-PAGE exhibited k-casein allele A in all the breeds. Moreover PCR+RFLP analysis also confirmed the presence of AA genotype in all the breeds. PCR-SSCP analysis of k-casein genetic variant showed the k-casein A and B allele in the analysed samples. The k-casein A allele was the dominant variant found in all analyzed breeds with frequencies ranging from 0.70 (Barbari) to 0.8 (Local MP). The variant B was most frequent in Barbari goats as compared to other breeds.

Keywords: Goat, κ-casein polymorphism, PCR-RFLP, SDS-PAGE, single strand conformation polymorphism

 

 

Indian Journal of Biotechnology

Vol 8, April 2009, pp 218-222

 

 

Acclimatization of neem microshoots adaptable to semi-sterile conditions

M Lavanya1, B Venkateshwarlu2 and B Poornasri Devi1*

1Department of Biosciences, Sri Sathya Sai University, Anantapur Campus, Anantapur 515 001, India

2Central Research Institute for Dryland Agriculture, Santoshnagar, Hyderabad 500 059, India

Received 25 March 2008; revised 2 September 2008; accepted 10 November 2008

The hardening of in vitro propagated microshoots of neem (Azadirachta indica A. Juss.) was carried out using 3 methods under semi-sterile conditions in low cost mini-polytunnels and a shade house. The percentage survival and rooting response was 16.25% in the first (2:1, v/v, sand and soil with 1′′ × 1′′ central cylindrical cocopeat plugs) and second method (1:1, v/v, cocopeat: bio-fertilizer), but was 100% in the third method (2:1, v/v, sand and soil with 1:1, v/v, cocopeat:biofertilizer and addition of Trichoderma viride. During the acclimatization process, the chlorophyll content in leaves gradually increased from 0.97 (stage I) to 1.35 (stage II), 1.56 (stage III) and 2.14 mg/g (stage IV), indicating a shift in the mode of nutrition from heterotrophic through myxotrophic to autotrophic. Similarly, the percentage water loss from the leaves of plantlets decreased from 90.38 (stage I) to 46.83% (stage IV), indicating stomatal development and progressive hardening. Ex vitro rooting and use of the bio-control agent could bring down the cost of production and make micropropagation of neem feasible and to be adopted as a rural enterprise.

Keywords: Bio-control agent, hardening, micropropagation, neem, Trichoderma

 

 

Indian Journal of Biotechnology

Vol 8, April 2009, pp 223-226

 

 

Micropropagation of Baliospermum montanum (Willd.) Muell. Arg.
A threatened medicinal plant

S Sasikumar, S Raveendar, A Premkumar, S Ignacimuthu* and P Agastian

Plant Biotechnology Unit, Entomology Research Institute, Loyola College, Chennai 600 034, India

Received 26 March 2008 ; revised 19 September 2008; accepted 25 November 2008

An efficient in vitro regeneration protocol was developed for Baliospermum montanum (Willd.) Muell. Arg., a wild threatened medicinal plant. The plants were regenerated from young nodal buds and shoot tips. The morphogenic frequency of shoot bud induction and shoot multiplication was significantly higher in nodal segments when compared to shoot tips. Maximum number of shoots (22.2 ± 0.84) with high frequency of shooting response (82%) was obtained in nodal explants cultured on MS medium fortified with 2.0 mg L-1 BAP. Maximum shoot height of 15.8 cm was achieved. Caulogenic effect of BAP was found to be significant compared to Kn. The excised shoots were cultured on MS medium with various concentrations and combinations of auxins for rooting. Maximum number of healthy rootlets (14.8 ± 2.07 cm) with 90% rooting response was observed due to a synergistic action of IBA (1.0 mg L-1) and IAA (0.5 mg L-1) on half strength MS basal medium. The regenerated plantlets were transferred to the natural habitat with 86.2% success.

Keywords: Baliospermum montanum, micropropagation, threatened plant

 

 

Indian Journal of Biotechnology

Vol 8, April 2009, pp 227-231

 

 

Efficacy of non-purine and purine cytokinins on shoot regeneration in vitro in sugarcane

Vandana Vinayak, Ashok K Dhawan* and V K Gupta**

CCS Haryana Agricultural University, Regional Research Station, Uchani, Karnal 131 001, India

Received 16 April 2008; revised 8 September 2008; accepted 15 November 2008

Effect of two non-purine cytokinins, TDZ [N-phenyl-N˘ - (1,2,3-thidiazol-5-yl) urea] and 4-CPPU [N- (2-chloro-4-pyridyl) N-phenylurea] and meta-topolin [6 (3-hydroxybenzylamino) purine] a biologically active aromatic compound, on shoot regeneration in sugarcane var. CoS 8436 and Co 1148 and S. officinarum clone ‘Gungera’ was observed at 15, 25 and 35 d. 4-CPPU significantly enhanced shoot regeneration in var. CoS 8436, but showed no effect or caused only a slight promotion in var. Co 1148 and clone ‘Gungera’. Meta-topolin was most effective in promoting shoot regeneration at 5 µM in var CoS 8436, at 1 µM in var Co 1148 and at 10 µM in in clone Gungera. The best response to TDZ was observed at 0.01 µM in vars CoS 8436 and Co 1148 and at 0.1 µM in clone Gungera. Thus, there was a clear varietal response of sugarcane to these chemicals. Clone ‘Gungera’ responded best to meta-topolin, CoS 8436 to 4-CPPU and Co 1148 to TDZ. Results obtained in this work clearly demonstrate TDZ induced shoot regeneration in sugarcane at unusually low concentration, such as 0.01 µM and 0.1 µM. Further, 4-CPPU, meta-topolin and TDZ simulate cytokinin activity and may thus provide a better substitute of cytokinins like BAP or Kn that are generally used in tissue culture.

Keywords: 4-CPPU, meta-toplin, TDZ, sugarcane, shoot regeneration, in vitro, growth regulators

 

 

Indian Journal of Biotechnology

Vol 8, April 2009, pp 232-235

 

 

Rapid clonal multiplication through in vitro axillary shoot proliferation of
Centella asiatica L.

K Karthikeyan*, C Chandran and S Kulothungan

Plant Tissue Culture Laboratory, P G and Research Department of Botany and Microbiology

A V V M Sri Pushpam College (Autonomous), Poondi 613 503, India

Received 13 February 2008; revised 22 September 2008; accepted 28 November 2008

Single nodal explants isolated from field grown plants of Centella asiatica, an important medicinal plant, when cultured for 4 wks on Murashige and Skoog’s (MS) medium containing different concentrations and combinations of BAP and Kn produced multiple shoots. A maximum of 15.24 shoots/node were produced after 30 d of culture in the presence of 2.0 mg/L BAP. Individual shoots (2-5 cm), when transferred onto full strength MS medium containing 1.5 mg/L IBA induced maximum number of roots (12.6). The rooted plants were successfully established in greenhouse condition after hardening.

Keywords: Centella asiatica, medicinal plant, multiple shoot, in vitro, clonal multiplication

 

 

Indian Journal of Biotechnology

Vol 8, April 2009, pp 236-239

 

 

In vitro propagation of Caralluma sarkariae Lavranos & Frandsen – An endemic and endangered medicinal plant

V Raja Sreelatha, S Sandhya Rani, P V Krishna Reddy,
M Naveen, A Ugraiah and T Pullaiah*

Department of Botany, Sri Krishnadevaraya University
Anantapur 515 003, India

Received 9 April 2008; revised 19 September 2008;
accepted 25 November 2008

An efficient protocol has been developed for in vitro propagation of Caralluma sarkariae Lavranos & Frandsen via mature internodal derived callus. Optimal callus was developed for regeneration from mature internodal explants on Murashige and Skoog (MS) basal medium supplemented with various concentrations of auxins. Maximum number of shoots were regenerated (65%) from the callus on MS medium supplemented with 6-benzyl aminopurine (BAP) 2.0 mg/L + kinetin (KN) 0.5 mg/L + naphthalene acetic acid (NAA) 0.5 mg/L. Individual elongated shoots were rooted on half strength MS medium containing NAA 0.1 mg/L. Regenerated plantlets with well developed shoots and roots were successfully transferred to soil.

Keywords: Callus cultures, Caralluma sarkariae, tissue culture

 

 

Indian Journal of Biotechnology

Vol 8, April 2009, pp 240-243

 

 

Microflora (fungal and bacterial) of selected terrestrial and marshy species of rhizosphere in response to spent wash treatments

 

P K Singh and K P Sharma*

Department of Botany, University of Rajasthan, Jaipur 302 004, India

Received 10 July 2007; revised 3 October 2008;
accepted 23 December 2008

In the present study, 16 fungal species were recorded in the rhizosphere of 4 terrestrial species [Acacia farnesiana (Linn.) Willd, A. leucophloea (Robx.) Willd, A. nilotica (Linn.) Del. and A. raddiana Savi] and 4 marshy species (Arundo donax Linn., Phragmites karka Steud, Typha angustata Bory & Chaub and Scirpus tuberosus Derf.) grown in tap water (control) and spent wash treatments (COD = 750-12,000 ppm). Their species richness was relatively higher (2-folds) in the spent wash treatments than their respective controls while their colony forming units were almost similar, exceeding in number to more than 300 units for a particular fungal species in the community. Aspergillus was the most dominant genus (4 species) followed by Fusarium (2 species) while the remaining 10 genera had one species each. The bacteria were Gram (+) bacilli and cocci arranged singly and in chains (also in bunches in cocci). In comparison to control, their CFU values were significantly higher (2-10-folds) in spent wash treatments, especially in the rhizosphere of marshy species.

Keywords: Fungi, bacteria, rhizosphere, terrestrial and marshy plant species, spent wash