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Background: Sugarcane is a tropical crop of major importance primarily for its high sucrose content. It is difficult to conserve it in the field or in vitro because of biotic and abiotic stresses. Cryopreservation of sugarcane germplasm... more
Background: Sugarcane is a tropical crop of major importance primarily for its high sucrose content. It is difficult to conserve it in the field or in vitro because of biotic and abiotic stresses. Cryopreservation of sugarcane germplasm is an appropriate approach for conserving its genetic diversity. Objective: This study was carried out to develop an efficient and practical cryopreservation protocol for sugarcane with high post-cryopreservation recovery. Materials and methods: Factors affecting regrowth after cryopreservation using the V cryo-plate method including preculture medium, size of shoot tips, sucrose concentration in loading solution, exposure time to PVS2, light conditions after liquid nitrogen exposure, presence and absence of alginate gel and recovery medium composition were studied. Results: Shoot tips with a length of 1.5 to 2.0 mm, precultured on semi-solid 1/2 MS medium for 1 day and semi-solid MS medium with 0.5 M sucrose for 1 day, treated with LS containing 2.0 M glycerol + 1.6 M sucrose for 30 min and exposed to PVS2 for 30 min showed maximum (100%) recovery after cryopreservation. It was also observed that removing the alginate gel and keeping the cultures in the dark for 7 days after cryopreservation significantly improved recovery. After optimizing the cryopreservation conditions using sugarcane variety Ni-1, 10 additional varieties were cryopreserved using the optimized protocol, with regrowth ranging from 56.7% to 100%. Conclusion: This study showed that V cryo-plate is an efficient and practical method for cryopreservation of sugarcane shoot tips in genebanks.
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A vitrification procedure using aluminium cryo-plates (V-Cryo-plate procedure) was successfully developed and adjusted for in vitro-grown mint (Mentha spp.) shoot tips. Shoots were cultured at 25°C on MS medium containing 0.088 M sucrose... more
A vitrification procedure using aluminium cryo-plates (V-Cryo-plate procedure) was successfully developed and adjusted for in vitro-grown mint (Mentha spp.) shoot tips. Shoots were cultured at 25°C on MS medium containing 0.088 M sucrose for 7 to 14 days after the last subculture. Shoot tips with a basal part (1-1.5 mm × 1 mm) were dissected from the shoots and precultured at 25°C for 1 day on the same medium. Precultured shoot tips were placed on aluminium cryo-plates with 10 wells and embedded in alginate gel. Osmoprotection was performed by immersing the cryo-plates for 30 min at 25 degree C in 25 ml pipetting reservoirs filled with loading solution (2 M glycerol + 0.8 M sucrose). For dehydration, the cryo-plates were transferred and immersed in 25 ml pipetting reservoirs filled with PVS2 for 20 min at 25 degree C. Then the cryo-plates were transferred in uncapped 2 ml cryotubes and directly plunged into liquid nitrogen. For rewarming, shoot tips attached to the cryo-plates were immersed in cryotubes containing 2 ml 1 M sucrose solution at room temperature. Using this procedure, regrowth of cryopreserved shoot tips of line 'Fukuyamajisei' reached over 90 percent. This protocol was successfully applied to 16 additional Mentha lines, with regrowth ranging from 73 percent to 100 percent. This V-Cryo-plate method will facilitate the cryostorage of mint germplasm in our genebank.
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A cryopreservation procedure using an aluminium cryo-plate was successfully developed using in vitro-grown Dalmatian chrysanthemum (Tanacetum cinerariifolium) shoot tips. Shoot cultures were cold-hardened at 5 degree C on MS medium... more
A cryopreservation procedure using an aluminium cryo-plate was successfully developed using in vitro-grown Dalmatian chrysanthemum (Tanacetum cinerariifolium) shoot tips. Shoot cultures were cold-hardened at 5 degree C on MS medium containing 0.5 M sucrose over a period of 20 to 40 days. Shoot tips with basal plate (1.0-1.5 x 1.0 mm) were dissected from shoot cultures and precultured at 5 degree C for 2 days on MS medium containing 0.5 M sucrose. Precultured shoot tips were placed on aluminium cryo-plates (7 mm x 37 mm x 0.5 mm) with 10 wells (diameter 1.5 mm, depth 0.75 mm) and embedded in alginate gel. Osmoprotection was performed by immersing the cryo-plates for 30 or 60 min in 25 ml pipetting reservoirs filled with loading solution (2 M glycerol + 1.4 M sucrose). For dehydration, the loading solution was replaced with PVS 7M vitrification solution (30 percent glycerol, 19.5 percent ethylene glycol and 0.6 M sucrose in liquid MS basal medium), which was applied for 40 min. After rapid immersion in liquid nitrogen, shoot tips attached to the cryo-plates were rewarmed by immersion in cryotubes containing 2 ml 1 M sucrose solution. Using this procedure, regrowth of cryopreserved shoot tips of line 28v-75 reached 77 degree. This protocol was successfully applied to six additional lines, with high regrowth percentages ranging from 65 to 90 percent. By contrast, the modified vitrification protocol tested as a reference produced only moderate regrowth percentages. This new method displays many advantages and will facilitate large scale cryostorage in genebank.
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Research Interests: Botany, Horticulture, Biology, Cryopreservation, Sucrose, and 5 moreGenebank, Liquid Nitrogen, Diospyros kaki, Shoot, and Kaki
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Wheat is one of the most important food crops, and its yield losses range from 20 to 100% due to leaf, stripe and stem rusts caused by Puccinia triticina, P. striiformis and P. graminis, respectively. New sources of resistance are... more
Wheat is one of the most important food crops, and its yield losses range from 20 to 100% due to leaf, stripe and stem rusts caused by Puccinia triticina, P. striiformis and P. graminis, respectively. New sources of resistance are strategically essential to limit the devastating effect of ever-evolving rust pathogens. In this study, we characterized 200 synthetic hexaploid wheats (SHWs) against all three rusts using rust pathotypes at seedling and adult plant stages in Queensland, Australia. We identified 57 accessions resistant to leaf rust, 77 to stripe rust and 69 accessions resistant to stem rust at the seedling stage. Ten SHWs were resistant to all three rusts, while 32 SHWs had dual resistance against leaf and stem rusts, and 28 SHWs had dual resistance to stripe and stem rust. We identified 24 SHWs carrying adult plant resistance (APR) for leaf, stripe and stem rust. The coefficient of correlation between yellow rust scores at seedling and adult plant stages were highest (r= ...
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Page 1. INTERNATIONAL JOURNAL OF AGRICULTURE & BIOLOGY 15608530/2004/065790791 http://www.ijab.org Heritability and Genetic Advance Estimates from Maize Genotypes in Shishi Lusht a Valley of Krakurm ...
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This research was planned to study the effect of various concentrations of BAP and sucrose on in vitro potato (Solanum tuberosum L.) microtuber induction. Explants from in vitro grown plants were cultured on Murashige and Skoog (1962)... more
This research was planned to study the effect of various concentrations of BAP and sucrose on in vitro potato (Solanum tuberosum L.) microtuber induction. Explants from in vitro grown plants were cultured on Murashige and Skoog (1962) medium supplemented with iron (150 ...
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Bitter gourd is one of the important cucurbits and highly liked among both farmers and consumers due to its high net return and nutritional value. However, being monoecious, it exhibits substantial variation in flower bearing pattern.... more
Bitter gourd is one of the important cucurbits and highly liked among both farmers and consumers due to its high net return and nutritional value. However, being monoecious, it exhibits substantial variation in flower bearing pattern. Plant growth regulators (PGRs) are known to influence crop phenology while gibberellic acid (GA3) is one of the most prominent PGRs that influence cucurbits phenology. Therefore, a field trial was conducted at University of Agriculture Faisalabad to evaluate the impact of a commercial product of gibberellic acid (GA3) on growth, yield and quality attributes of two bitter gourd (Momordica charantiaL.) cultivars. We used five different concentrations (0.4 g, 0.6 g, 0.8 g, 1.0 g, and 1.2 g per litre) of commercial GA3 product (Gibberex, 10% Gibberellic acid). Results showed that a higher concentration of gibberex (1.0 and 1.20 g L−1 water) enhanced the petiole length, intermodal length, and yield of bitter gourd cultivars over control in Golu hybrid and F...
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The aim of the present investigation was to determine the effect of salt stress on the physiology of twelve genotypes of tomato (Solanum lycopersicum L.), to study the genetic diversity among these genotypes and to transfer salt tolerant... more
The aim of the present investigation was to determine the effect of salt stress on the physiology of twelve genotypes of tomato (Solanum lycopersicum L.), to study the genetic diversity among these genotypes and to transfer salt tolerant gene in the selected genotype through Agrobacterium mediated gene transformation method.The seeds of different tomato genotypes were obtained from Ayub Agricultural Research Institute (AARI), Faisalabad and from Horticulture Research institute (HRI), NARC, Islamabad. First experiment comprises study of plant growth, ionic content, ABA content of shoot and root and proline content at four different NaCl salinity level.The genotypes Avinash, NTH242 and Gol performed better at higher salt concentration as indicated by less inhibition of plant height, shoot and root fresh weight and shoot and root dry weight.There was less translocation of Na+ to shoot but greater accumulation of K+ and Ca2+ concomitant with higher production of osmoregulant (proline) a...
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Sugarcane is a tropical crop of major importance primarily for its high sucrose content. It is difficult to conserve it in the field or in vitro because of biotic and abiotic stresses. Cryopreservation of sugarcane germplasm is an... more
Sugarcane is a tropical crop of major importance primarily for its high sucrose content. It is difficult to conserve it in the field or in vitro because of biotic and abiotic stresses. Cryopreservation of sugarcane germplasm is an appropriate approach for conserving its genetic diversity. This study was carried out to develop an efficient and practical cryopreservation protocol for sugarcane with high post-cryopreservation recovery. Factors affecting regrowth after cryopreservation using the V cryo-plate method including preculture medium, size of shoot tips, sucrose concentration in loading solution, exposure time to PVS2, light conditions after liquid nitrogen exposure, presence and absence of alginate gel and recovery medium composition were studied. Shoot tips with a length of 1.5 to 2.0 mm, precultured on semi-solid 1/2 MS medium for 1 day and semi-solid MS medium with 0.5 M sucrose for 1 day, treated with LS containing 2.0 M glycerol + 1.6 M sucrose for 30 min and exposed to P...
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Cryopreservation protocols using aluminium plates were successfully developed by our team. The size of an aluminum cryo-plate used is 7 mm × 37 mm × 0.5 mm with 10 to 16 wells fitting in 2 ml cryotube. The cryo-plates were custom-made by... more
Cryopreservation protocols using aluminium plates were successfully developed by our team. The size of an aluminum cryo-plate used is 7 mm × 37 mm × 0.5 mm with 10 to 16 wells fitting in 2 ml cryotube. The cryo-plates were custom-made by Taiyo Nippon Sanso Corp. Tokyo, Japan. The cryopreservation protocols developed using this cryo-plate are V cryo-plate method based on PVS2 dehydration of explants on the plates, and D cryo-plate method based on air dehydration. These procedures include the following steps: preparation of material to be cryopreserved, preconditioning, excision, preculture, mounting the shoot tips on cryo-plate, osmoprotection, dehydration, storage and regeneration. Both methods are efficient, ensures rapid cooling and warming rates, and reduce damage to explants before cryopreservation by attached shoot tips on cryo-plate. Also, they were developed with the aim of limiting the problems linked to operator skills and simplifying the procedure. The V cryo-plate method ...
Broad-sense heritabilty, coefficients of variability and genetic advance values were computed for days taken to tasseling, number of days taken to silking, plant height, ear length, number of kernel rows ear-1, number of kernels row-1,... more
Broad-sense heritabilty, coefficients of variability and genetic advance values were computed for days taken to tasseling, number of days taken to silking, plant height, ear length, number of kernel rows ear-1, number of kernels row-1, 100-grain weight and grain yield plant -1 . Low, medium and high estimates of broad sense heritability were found in different plant characters under study. Highest heritability estimates were found in grain yield plant-1 (0.993) and by plant height (0.990). Values of genetic advance ranged between 43.80 for grain yield plant -1 to 1.33 for number of kernel rows ear -1 . Greater magnitude of broad sense heritability coupled with higher genetic advance in characters under study provided the evidence that these plant parameters were under the control of additive genetic effects. Indicating that selection should lead to a fast genetic improvement of the material
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A vitrification procedure using aluminium cryo-plates (V-Cryo-plate procedure) was successfully developed and adjusted for in vitro-grown mint (Mentha spp.) shoot tips. Shoots were cultured at 25°C on MS medium containing 0.088 M sucrose... more
A vitrification procedure using aluminium cryo-plates (V-Cryo-plate procedure) was successfully developed and adjusted for in vitro-grown mint (Mentha spp.) shoot tips. Shoots were cultured at 25°C on MS medium containing 0.088 M sucrose for 7 to 14 days after the last subculture. Shoot tips with a basal part (1-1.5 mm × 1 mm) were dissected from the shoots and precultured at 25°C for 1 day on the same medium. Precultured shoot tips were placed on aluminium cryo-plates with 10 wells and embedded in alginate gel. Osmoprotection was performed by immersing the cryo-plates for 30 min at 25 degree C in 25 ml pipetting reservoirs filled with loading solution (2 M glycerol + 0.8 M sucrose). For dehydration, the cryo-plates were transferred and immersed in 25 ml pipetting reservoirs filled with PVS2 for 20 min at 25 degree C. Then the cryo-plates were transferred in uncapped 2 ml cryotubes and directly plunged into liquid nitrogen. For rewarming, shoot tips attached to the cryo-plates were ...
Research Interests:
A cryopreservation procedure using an aluminium cryo-plate was successfully developed using in vitro-grown Dalmatian chrysanthemum (Tanacetum cinerariifolium) shoot tips. Shoot cultures were cold-hardened at 5 degree C on MS medium... more
A cryopreservation procedure using an aluminium cryo-plate was successfully developed using in vitro-grown Dalmatian chrysanthemum (Tanacetum cinerariifolium) shoot tips. Shoot cultures were cold-hardened at 5 degree C on MS medium containing 0.5 M sucrose over a period of 20 to 40 days. Shoot tips with basal plate (1.0-1.5 x 1.0 mm) were dissected from shoot cultures and precultured at 5 degree C for 2 days on MS medium containing 0.5 M sucrose. Precultured shoot tips were placed on aluminium cryo-plates (7 mm x 37 mm x 0.5 mm) with 10 wells (diameter 1.5 mm, depth 0.75 mm) and embedded in alginate gel. Osmoprotection was performed by immersing the cryo-plates for 30 or 60 min in 25 ml pipetting reservoirs filled with loading solution (2 M glycerol + 1.4 M sucrose). For dehydration, the loading solution was replaced with PVS 7M vitrification solution (30 percent glycerol, 19.5 percent ethylene glycol and 0.6 M sucrose in liquid MS basal medium), which was applied for 40 min. After ...