Plant Tissue Culture Laboratory

Karachi, Pakistan

Plant Tissue Culture Laboratory

Karachi, Pakistan
SEARCH FILTERS
Time filter
Source Type

Das J.,Plant Tissue Culture Laboratory | Das J.,Gauhati University | Mao A.A.,Plant Tissue Culture Laboratory | Handique P.J.,Gauhati University
Acta Physiologiae Plantarum | Year: 2013

A reproducible and efficient callus-mediated shoot regeneration system was developed for the large-scale production of Valeriana jatamansi Jones., a highly medicinal plant species of global pharmaceutical importance. Effect of Murashige and Skoog (MS) medium supplemented with different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D), α-naphthaleneacetic acid (NAA) and indole-3-butyric acid (IBA) on callus induction and production of valepotriates accumulation was studied by using different explants. In V. jatamansi, the degree of callus induction varied significantly depending on explants type and the growth regulators used. Among different explants used, rhizomes have the highest callus induction potential followed by leaf. The callus induction frequency was found to be optimum in rhizome explants on media supplemented with 0. 5 mg/l 2,4-D. The regenerative ability of proliferated compact calli was studied by the application of cytokinins alone and in combination with auxin. MS medium fortified with 0. 75 mg/l thidiazuron in combination with 0. 5 mg/l NAA showed the highest regeneration frequency (88. 6 %) and produced the maximum number of shoot buds (15. 20 ± 0. 20) capable of growing into single plants. Vigorous callus obtained from MS medium supplemented with different concentrations of 2,4-D, NAA and IBA were used for industrially important valepotriates [acevaltrate (ACE), valtrate (VAL) and didrovaltrate (DID)] analysis. High performance liquid chromatography analysis of callus revealed that medium with 2,4-D (1 mg/l) was found responsible for increasing ACE and DID yield, whereas VAL production was higher in case of medium supplemented with NAA (1 mg/l). However, the accumulation of valepotriates in callus decreased in logarithmic phase after 8 weeks. IBA was not beneficial for the valepotriate synthesis, as it helped to accumulate significantly lower concentration of ACE, VAL and DID. Micropropagated plantlets with well-developed root system were successfully acclimatized in greenhouse condition, in root trainers containing garden soil with a survival frequency of 100 %. As Indian valerian is a highly traded medicinal plant due to extensive use of its industrially important secondary metabolites, the present system can be utilized to obtain mass multiplication of the species as well as for the stable biomass and continuous valepotriate production for the pharmaceutical industries. © 2012 Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków.


De C. Silva R.,Federal University of Amazonas | Camillo J.,University of Brasilia | Scherwinski-Pereira J.E.,Plant Tissue Culture Laboratory
Revista de Biologia Tropical | Year: 2012

Actually, the germplasm of Jatropha spp. is conserved as whole plants in field collections. Under this storage method, the genetic resources are exposed to disease, pest and natural hazards such as human error, drought and weather damage. Besides, field genebanks are costly to maintain and with important requirements of trained personnel. Thus, the development of efficient techniques to ensure its safe conservation and regeneration is therefore of paramount importance. In this work we describe a method for Jatropha curcas seeds cryoexposure and seedling recovery after thawed. In a first experiment, an efficient protocol for in vitro plant recovery was carried out using zygotic embryo or seeds with or without coat. In a second experiment, desiccated seeds with or without coat were exposed to liquid nitrogen and evaluated after cryoexposure. Germination percentages were variable among treatments, and seeds demonstrated tolerance to liquid nitrogen exposure under certain conditions. Seeds of J. curcas presented up to 99.6% germination after seed coat removal. Seeds with coat cultured in vitro did not germinate, and were 60% contaminated. The germination of the zygotic embryos was significantly higher in the 1/2 MS medium (93.1%) than in WPM medium (76.2%), but from zygotic embryo, abnormal seedlings reached up to 99%. Seeds with coat exposed to liquid nitrogen showed 60% germination in culture after coat removal with good plant growth, and seeds cryopreserved without coat presented 82% germination, but seedlings showed a reduced vigor and a significant increase in abnormal plants. Seeds cultured in vitro with coat did not germinate, independently of cryoexposure or not. This study reports the first successful in vitro seedling recovery methodology for Jatropha curcas seeds, after a cryopreservation treatment, and is recommended as an efficient procedure for in vitro plant recovery, when seeds are conserved in germplasm banks by low or cryotemperatures.


Ikram-Ul-Haq,University of Sindh | Dahri A.M.,Plant Tissue Culture Laboratory | Dahot M.U.,University of Sindh | Parveen N.,University of Sindh | And 2 more authors.
African Journal of Biotechnology | Year: 2010

Negative impact of salinity on plant germination is significant because of abundance of Na+ in culture medium, which causes growth inhibition. Effect of salinity (NaCl) in the presence of proline was assessed in rice (Oryza sativa L.) variety Khushbo-95 at seedling stage. Seeds were cultured on MS0 (MS basal medium), MS1 (MS0 + 100 mM NaCl) and MS2 (MS1 + 5 mM proline) for 20 days. Seedlings and its biomass decreased in saline culture. Similarly, total protein and sugar contents also decreased, while reducing sugars and proline contents increased. These parameters were observed to be slightly adverse in cultures supplemented with proline (MS2) and NaCl (MS2). Among cultures, leaf demography (cell size) was affected significantly; this may be the reflection of accumulation of proline, Na+ and Clsup- and exclusion of K+ in developed rice seedlings. © 2010 Academic Journals.


Hamwieh A.,International Center for Agricultural Research in the Dry Areas | Farah J.,University of Aleppo | Moussally S.,University of Aleppo | Al-Sham'Aa K.,International Center for Agricultural Research in the Dry Areas | And 7 more authors.
Acta Horticulturae | Year: 2010

Date Palm is a major environmental and economic factor in arid climates in many countries around the world. Microsatellite markers have been proven to be very powerful in plant genome analysis because they are locus-specific, codominant, highly polymorphic and highly reproducible. In date palm only few microsatellite markers have been developed so far. Recently, the Cornell Medical College in Qatar issued a draft assembly of the date palm genome ('Khalas') generated by whole genome shotgun next generation DNA sequencing. In this paper, we analyzed the microsatellite motifs across the date palm genome. The results indicated that the most abundant type of microsatellite repeats are dinucleotide repeats (52442 motifs) followed by trinucleotide (28503 motifs) and pentanucleotide repeats (12873 motifs). The frequencies of tetra-nucleotide and hexa-nucleotide repeats were less across the genome (5555 and 5810 motifs, respectively). The most common type of dinucleotide repeat was GA (48.7%) followed by AT (37%). Out of 28645 trinucleotide repeats, TAA and GAA repeats were the most abundant repeats (28.1 and 27.1%) respectively. More than 1090 new microsatellite markers could be designed. The primary test for 50 primer pairs revealed that 28 (56%) were functional and 19 (38%) yielded polymorphic PCR products. We wish that the results of our study will be a starting point for researchers making use of the markers for genetic mapping and diversity analysis of date palm.


Waman A.A.,Plant Tissue Culture Laboratory | Sathyanarayana B.N.,Plant Tissue Culture Laboratory | Umesha K.,Plant Tissue Culture Laboratory | Gowda B.,University of Agricultural Sciences, Bangalore | And 3 more authors.
Journal of Applied Horticulture | Year: 2011

Withania somnifera (L.) Dunal., one of the 32 prioritized medicinal plants of India, is well known for its importance in the Ayurveda system of medicine. Attempt was made to establish an efficient plant regeneration protocol for a commercial cultivar 'Poshita' and to acclimatize the plantlets ex vitro, so as to reduce the cost. Results revealed that shoot induction was possible only after an intervening callus phase, irrespective of the concentration of growth regulators present in the nutrient pool. Nodal explant cultured on MS media supplemented with 1 mg L-1 BAP + 0.5 mg L-1 NAA showed superiority in callus induction capacity over epicotyls and leaves. Nodal segments when cultured on a media containing BAP alone could induce shoots in cent per cent explants. Highest number of shoots (5.8) was obtained in media containing 2 mg L-1 BAP. Number of adventitious buds was found to be maximum (13) with epicotyls explant and 1 mg L-1 BAP combination. Nodal explants cultured on high concentration of BAP (4 mg L-1) showed highest incidence of malformed shoots (4.3). A total of 66.7 % plantlets could root and establish ex vitro even without auxin treatment and survival rate increased (87.5%) with increase in IBA concentration to 500 mg L-1. The present protocol can be exploited on a commercial scale to obtain maximum benefits from the improved cultivar. Furthermore, ex vitro hardening can help to reduce the cost of production and thereby make the tissue culture industry more profitable.


Naqvi B.,Plant Tissue Culture Laboratory | Tariq Y.,Plant Tissue Culture Laboratory
Pakistan Journal of Scientific and Industrial Research Series B: Biological Sciences | Year: 2011

This study describes an effective and reproducible protocol for the mass multiplication of marigold (Tagetes erecta L.) for commercial purpose. Twenty five different combinations of BAP, IAA, GA3 and AgNO3 were added to the basal MS medium to culture marigold explants. The highest mean number (4.83±0.49) and length (5.28 cm ±1.06) of healthy shoots per explant was observed in media supplemented with 2 mg/L BAP along with 2 mg/L IAA. When these shoots were sub-cultured for root development, the maximum number (17.08±2.44) and length (13.67 cm ±0.98) of roots were produced in media supplemented with 4mg/L BAP and 2 mg/L IAA. Addition of gibberellic acid and AgNO3 did not have any significant effect on shoot proliferation and root development of marigold.


Giri D.,Plant Tissue Culture Laboratory | Tamta S.,Plant Tissue Culture Laboratory | Pandey A.,Plant Tissue Culture Laboratory
Journal of Medicinal Plants Research | Year: 2010

The objective of this study was to compile a review note on distribution, medicinal properties, trade value and problems associated with an important medicinal plant, Hedychium spicatum Buch-Ham ex Sm (Zingiberaceae). The underground part (rhizome) is useful in preparation of indigenous medicine like the crude extract of the rhizome has been used in preparation of an anti cancerous drug. Essential oil (commercially known as 'Kapur Kachri Oil') from rhizomes showed antimicrobial activity against both Gram-positive and Gram-negative bacteria. On the basis of comprehensive study of literature it has been found that this species is commercially exploited from its natural habitat. Hence, prioritization needs to be done for propagation and conservation of H. spicatum. © 2010 Academic Journals.


Carmona-Martin E.,Plant Tissue Culture Laboratory | Regalado J.J.,Plant Tissue Culture Laboratory | Raghavan L.,Sugarcane Breeding Institute | Encina C.L.,Plant Tissue Culture Laboratory
Plant Cell, Tissue and Organ Culture | Year: 2015

Octoploid genotypes of Asparagus officinalis L. cv. ‘Morado de Huetor’, a tetraploid Spanish landrace, were successfully induced by treating in vitro rhizome buds explants with colchicine. Pulses during 24 h with different concentrations of colchicine are able to induce polyploid plants, colchicine applied at 0.1 % induce a 7 % of octoploid genotypes with a stable ploidy level and a 3.5 % of mixoploid genotypes. The maximum rate of mixoploid genotypes (26 %) was obtained with 0.3 % colchicine. The plant survival and rooting rates of explants decrease for increasing doses of colchicine. All octoploid genotypes were micropropagated, rooted, transplanted and successfully acclimatized (100 %) in a glasshouse. The four octoploid plants recovered show significantly better agronomical parameters such as spear diameter, canopy area and shoot length than the original tetraploid plants and histological studies confirm the size increase of octoploid cells respect the original tetraploid cells. © 2014, Springer Science+Business Media Dordrecht.

Loading Plant Tissue Culture Laboratory collaborators
Loading Plant Tissue Culture Laboratory collaborators