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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

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. Source

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

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. Source

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

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. Source

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

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. Source

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

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. Source

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