In order to understand the chromosome morphology and structural variation, somatic chromosomes of... more In order to understand the chromosome morphology and structural variation, somatic chromosomes of nine accessions of grass pea (Lathyrus sativus L.) viz., ‘Pusa 24’, ‘Ratan’, ‘Pratik’, ‘IC120455’, ‘IC120500’, ‘IC120505’, ‘IC345392’, ‘IC345401’, and ‘IC345403’ were studied using young root tips. The study revealed variation not only in physical dimension and morphology of the chromosomes but also in the position of the primary and secondary constrictions. Although all accessions were diploid (2n=14), variations in karyotype formula, chromosome length, total haploid complement, arm ratio and centromeric index were observed. The secondary constriction was observed in at least one of the metacentric chromosomes of ‘Pusa 24’, ‘IC120500’, ‘IC120505’, ‘IC345392’ and ‘IC345403’. Telocentric chromosomes were absent in all the accessions while variable numbers of sub-metacentric and acrocentric chromosomes were observed in these accessions. These chromosomes differed in their lengthwise positions in the karyotype. Total length of the largest chromosomes did not differ significantly among accessions. However, chromosomes with satellites were longer. Total length of smallest chromosome was variable among the accessions. The total haploid chromosome length (TCL) varied from 5.11 µm in ‘IC345403’ to 7.30 µm in ‘IC345401’. The arm ratio and centromeric index value ranged from 1.21 in ‘IC120455’ to 1.44 in ‘IC120505’ and 40.94 in ‘IC120505’ to 45.19 in ‘IC120455’, respectively. These results clearly showed structural variation in karyotype at genotypic level in grass pea.
Abstract Agrobacterium- mediated genetic transformation approach allows for introducing novel gen... more Abstract Agrobacterium- mediated genetic transformation approach allows for introducing novel genes in cotton ( Gossypium hirsutum L.). Development of efficient regeneration and transformation protocol is very important for recalcitrant plants like cotton. In the present study, five-day-old germinated mature embryo parts especially embryonic axis, hypocotyl and plumule of cotton ‘Lashata’ were wounded and inoculated with A. tumefaciens strain GV2260 harbouring plasmid p35S-GUS-INT. The binary plasmid p35S-GUS-INT contains neomycin phosphotransferase II ( NPTII ) gene driven by nopaline synthase ( NOS ) promoter and β-glucuronidase ( GUS ) gene controlled by cauliflower mosaic virus (CaMV) 35S promoter. After inoculation, explants were co-cultivated in liquid and agar-solidified MS medium for 48 h in dark condition. Agar-solidified co-cultivation medium increased transformation frequency as compared to liquid medium. The putative primary transformants were confirmed with histochemical GUS assay, PCR and RT-PCR. However, comparing the three culture explant, the embryonic axis explants had significant difference with embryonic hypocotyl and plumule explants in terms of number and percentage (%) of GUS, PCR and RT-PCR positive plant. The total transformation efficiency was recorded as 3% with varying GUS expression levels.
The rich genetic diversity existing within exotic, indigenous, and diverse germplasm lays the fou... more The rich genetic diversity existing within exotic, indigenous, and diverse germplasm lays the foundation for the continuous improvement of crop cultivars. The composite collection has been suggested as a gateway to identifying superior germplasm for use in crop improvement programs. Here, a chickpea global composite collection was evaluated at five locations in India over two years for five agronomic traits to identify agronomically superior accessions. The desi, kabuli, and intermediate types of chickpea accessions differed significantly for plant height (PLHT) and 100-seed weight (100 SW). In contrast, the intermediate type differed substantially from kabuli for days to maturity (DM). Several highly significant trait correlations were detected across different locations. The most stable and promising accessions from each of the five locations were prioritised based on their superior performance over the best-performing check cultivar. Accordingly, the selected germplasm accessions...
WOS: 000436522600048Potato (Solanum tuberosum L.) is highly demandable world food crop and has be... more WOS: 000436522600048Potato (Solanum tuberosum L.) is highly demandable world food crop and has become a staple food in many parts of the world including Turkey, India and China. Potato under in vitro culture condition is recalcitrant and genotype dependent. Therefore, we tested several factors to enhance regeneration and genetic transformation efficiency of Turkish potato genotypes. The influence of physical factors and conditions in Agrobacterium mediated transformation of the potato leaves was tested. Using the best combination (10 mg L-1 ZR, 10 mg L-1 NAA) with Agrobacterium-mediated transformation, we regenerated independent putative transformants 61.6% of the total explants cultured. GUS and nptII positive plants were confirmed by molecular analysis (PCR) that showed transformation rate of 24.16%. The protocol described here is simple, efficient, and produces transgenic plant in 7-8 weeks after inoculation with Agrobacterium bacterium.Scientific and Technological Research Council of Turkey (TUBITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK)The present work is partially supported by the Scientific and Technological Research Council of Turkey (TUBITAK). The authors would like to thank Prof. Dr. Gungor Yilmaz (University of Gaziosmanpasa, Department of Field Crops) and Dr. Huseyin Onaran (Nigde Potato Research Institute) for providing the plant material. The authors would like to thank Dr Jitendra Kumar (Department of Plant Pathology, University of Minnesota, USA) for valuable suggestion and improvement of the present manuscript
Grass pea (Lathyrus sativus L.) is highly recalcitrant and difficult to regenerate under in vitro... more Grass pea (Lathyrus sativus L.) is highly recalcitrant and difficult to regenerate under in vitro conditions. An efficient and reliable protocol was developed for induction of effective in vitro regeneration of grass pea using embryos with two cotyledons. The maximum shoot regeneration frequency of 89.80% and number of 15.83 shoots per explant was induced on MS medium containing 2 mg/l BAP and 1 mg/l NAA. The mean shoot length decreased proportionately with each increase in TDZ concentration irrespective of the IBA. The regenerated shoots produced roots on MS medium containing 0.5, 1.0 and 2.0 mg/l IBA. However, rooted plantlets flowered in the rooting media.
In Vitro Cellular & Developmental Biology - Plant, 2008
Leaf piece explants of five Brassica juncea (L.) Czern. cultivars were transformed with an Agroba... more Leaf piece explants of five Brassica juncea (L.) Czern. cultivars were transformed with an Agrobacterium tumefaciens strain EHA105 harboring the plasmid pCAMBIA1301, which contains the β-glucuronidase (uidA) and hygromycin phosphotransferase (hpt) genes under the control of cauliflower mosaic virus 35S (CaMV35S) promoter. Transgenic plants were regenerated on Murashige and Skoog (MS) medium fortified with 8.87 µM 6-benzylaminopurine, 0.22 µM 2,4-dichlorophenoxyacetic acid, and
ODAP (N-oxalyl-L-2,3-diaminopropionic acid) is present in the seeds of grass pea. In this study, ... more ODAP (N-oxalyl-L-2,3-diaminopropionic acid) is present in the seeds of grass pea. In this study, variation of total ODAP accumulation in leaves throughout the crop growth starting from 40 days after sowing to maturity, and the distribution pattern of ODAP in different plant parts including the seeds at the mature stage was analyzed. Five grass pea accessions were evaluated for two subsequent growing seasons in one location of ICARDA, Aleppo (Syria). The results found that the rate of accumulation of total ODAP varied during plant development. Increased rates of synthesis were noticed in young leaves of grass pea. The highest total ODAP content in leaves was noted in the early growth stage (40–50 days after sowing). Mean total ODAP content in leaves ranged from 0.17 to 0.96 percent during 2010–2011 and from 0.19 to 1.28 percent during 2011–2012. During maturity, the total ODAP content was lowest in the seeds than in leaves, stems, pod cover, seed coat, and cotyledons. The ranges of t...
WOS:000626629800014In the present study, effect of different regeneration media (R) and combinati... more WOS:000626629800014In the present study, effect of different regeneration media (R) and combination (R1 = MS medium, R2 = MS Salts + B5 vitamins, R3 = MS salts + SH vitamins, R4 = SH medium, R5 = SH salts + MS vitamins, R6 = SH salts + B5 vitamins) and various carbon sources (glucose, sucrose and fructose) was investigated on efficient in vitro shoot multiplication of Riccia fluitans. The media composition significantly affected (P <= 0.05) length, width, area, fresh and dry weight of clumps on multiplied plants. The R5 medium produced highest length (2.58 cm), width (2.16 cm), area (5.57 cm(2)), fresh weight (402.6 g) and dry weight (37.6 g) on regenerated clumps that showed healthier and green plant on culture medium. However, the regenerated clumps diameter was also significantly influenced (P <= 0.05) by the type and concentration of sugar (carbon sources) used. The highest clump diameter (7.27 cm) was obtained on R5 medium supplemented with 1.5% sucrose. Whereas, the high...
In order to understand the chromosome morphology and structural variation, somatic chromosomes of... more In order to understand the chromosome morphology and structural variation, somatic chromosomes of nine accessions of grass pea (Lathyrus sativus L.) viz., ‘Pusa 24’, ‘Ratan’, ‘Pratik’, ‘IC120455’, ‘IC120500’, ‘IC120505’, ‘IC345392’, ‘IC345401’, and ‘IC345403’ were studied using young root tips. The study revealed variation not only in physical dimension and morphology of the chromosomes but also in the position of the primary and secondary constrictions. Although all accessions were diploid (2n=14), variations in karyotype formula, chromosome length, total haploid complement, arm ratio and centromeric index were observed. The secondary constriction was observed in at least one of the metacentric chromosomes of ‘Pusa 24’, ‘IC120500’, ‘IC120505’, ‘IC345392’ and ‘IC345403’. Telocentric chromosomes were absent in all the accessions while variable numbers of sub-metacentric and acrocentric chromosomes were observed in these accessions. These chromosomes differed in their lengthwise positions in the karyotype. Total length of the largest chromosomes did not differ significantly among accessions. However, chromosomes with satellites were longer. Total length of smallest chromosome was variable among the accessions. The total haploid chromosome length (TCL) varied from 5.11 µm in ‘IC345403’ to 7.30 µm in ‘IC345401’. The arm ratio and centromeric index value ranged from 1.21 in ‘IC120455’ to 1.44 in ‘IC120505’ and 40.94 in ‘IC120505’ to 45.19 in ‘IC120455’, respectively. These results clearly showed structural variation in karyotype at genotypic level in grass pea.
Abstract Agrobacterium- mediated genetic transformation approach allows for introducing novel gen... more Abstract Agrobacterium- mediated genetic transformation approach allows for introducing novel genes in cotton ( Gossypium hirsutum L.). Development of efficient regeneration and transformation protocol is very important for recalcitrant plants like cotton. In the present study, five-day-old germinated mature embryo parts especially embryonic axis, hypocotyl and plumule of cotton ‘Lashata’ were wounded and inoculated with A. tumefaciens strain GV2260 harbouring plasmid p35S-GUS-INT. The binary plasmid p35S-GUS-INT contains neomycin phosphotransferase II ( NPTII ) gene driven by nopaline synthase ( NOS ) promoter and β-glucuronidase ( GUS ) gene controlled by cauliflower mosaic virus (CaMV) 35S promoter. After inoculation, explants were co-cultivated in liquid and agar-solidified MS medium for 48 h in dark condition. Agar-solidified co-cultivation medium increased transformation frequency as compared to liquid medium. The putative primary transformants were confirmed with histochemical GUS assay, PCR and RT-PCR. However, comparing the three culture explant, the embryonic axis explants had significant difference with embryonic hypocotyl and plumule explants in terms of number and percentage (%) of GUS, PCR and RT-PCR positive plant. The total transformation efficiency was recorded as 3% with varying GUS expression levels.
The rich genetic diversity existing within exotic, indigenous, and diverse germplasm lays the fou... more The rich genetic diversity existing within exotic, indigenous, and diverse germplasm lays the foundation for the continuous improvement of crop cultivars. The composite collection has been suggested as a gateway to identifying superior germplasm for use in crop improvement programs. Here, a chickpea global composite collection was evaluated at five locations in India over two years for five agronomic traits to identify agronomically superior accessions. The desi, kabuli, and intermediate types of chickpea accessions differed significantly for plant height (PLHT) and 100-seed weight (100 SW). In contrast, the intermediate type differed substantially from kabuli for days to maturity (DM). Several highly significant trait correlations were detected across different locations. The most stable and promising accessions from each of the five locations were prioritised based on their superior performance over the best-performing check cultivar. Accordingly, the selected germplasm accessions...
WOS: 000436522600048Potato (Solanum tuberosum L.) is highly demandable world food crop and has be... more WOS: 000436522600048Potato (Solanum tuberosum L.) is highly demandable world food crop and has become a staple food in many parts of the world including Turkey, India and China. Potato under in vitro culture condition is recalcitrant and genotype dependent. Therefore, we tested several factors to enhance regeneration and genetic transformation efficiency of Turkish potato genotypes. The influence of physical factors and conditions in Agrobacterium mediated transformation of the potato leaves was tested. Using the best combination (10 mg L-1 ZR, 10 mg L-1 NAA) with Agrobacterium-mediated transformation, we regenerated independent putative transformants 61.6% of the total explants cultured. GUS and nptII positive plants were confirmed by molecular analysis (PCR) that showed transformation rate of 24.16%. The protocol described here is simple, efficient, and produces transgenic plant in 7-8 weeks after inoculation with Agrobacterium bacterium.Scientific and Technological Research Council of Turkey (TUBITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK)The present work is partially supported by the Scientific and Technological Research Council of Turkey (TUBITAK). The authors would like to thank Prof. Dr. Gungor Yilmaz (University of Gaziosmanpasa, Department of Field Crops) and Dr. Huseyin Onaran (Nigde Potato Research Institute) for providing the plant material. The authors would like to thank Dr Jitendra Kumar (Department of Plant Pathology, University of Minnesota, USA) for valuable suggestion and improvement of the present manuscript
Grass pea (Lathyrus sativus L.) is highly recalcitrant and difficult to regenerate under in vitro... more Grass pea (Lathyrus sativus L.) is highly recalcitrant and difficult to regenerate under in vitro conditions. An efficient and reliable protocol was developed for induction of effective in vitro regeneration of grass pea using embryos with two cotyledons. The maximum shoot regeneration frequency of 89.80% and number of 15.83 shoots per explant was induced on MS medium containing 2 mg/l BAP and 1 mg/l NAA. The mean shoot length decreased proportionately with each increase in TDZ concentration irrespective of the IBA. The regenerated shoots produced roots on MS medium containing 0.5, 1.0 and 2.0 mg/l IBA. However, rooted plantlets flowered in the rooting media.
In Vitro Cellular & Developmental Biology - Plant, 2008
Leaf piece explants of five Brassica juncea (L.) Czern. cultivars were transformed with an Agroba... more Leaf piece explants of five Brassica juncea (L.) Czern. cultivars were transformed with an Agrobacterium tumefaciens strain EHA105 harboring the plasmid pCAMBIA1301, which contains the β-glucuronidase (uidA) and hygromycin phosphotransferase (hpt) genes under the control of cauliflower mosaic virus 35S (CaMV35S) promoter. Transgenic plants were regenerated on Murashige and Skoog (MS) medium fortified with 8.87 µM 6-benzylaminopurine, 0.22 µM 2,4-dichlorophenoxyacetic acid, and
ODAP (N-oxalyl-L-2,3-diaminopropionic acid) is present in the seeds of grass pea. In this study, ... more ODAP (N-oxalyl-L-2,3-diaminopropionic acid) is present in the seeds of grass pea. In this study, variation of total ODAP accumulation in leaves throughout the crop growth starting from 40 days after sowing to maturity, and the distribution pattern of ODAP in different plant parts including the seeds at the mature stage was analyzed. Five grass pea accessions were evaluated for two subsequent growing seasons in one location of ICARDA, Aleppo (Syria). The results found that the rate of accumulation of total ODAP varied during plant development. Increased rates of synthesis were noticed in young leaves of grass pea. The highest total ODAP content in leaves was noted in the early growth stage (40–50 days after sowing). Mean total ODAP content in leaves ranged from 0.17 to 0.96 percent during 2010–2011 and from 0.19 to 1.28 percent during 2011–2012. During maturity, the total ODAP content was lowest in the seeds than in leaves, stems, pod cover, seed coat, and cotyledons. The ranges of t...
WOS:000626629800014In the present study, effect of different regeneration media (R) and combinati... more WOS:000626629800014In the present study, effect of different regeneration media (R) and combination (R1 = MS medium, R2 = MS Salts + B5 vitamins, R3 = MS salts + SH vitamins, R4 = SH medium, R5 = SH salts + MS vitamins, R6 = SH salts + B5 vitamins) and various carbon sources (glucose, sucrose and fructose) was investigated on efficient in vitro shoot multiplication of Riccia fluitans. The media composition significantly affected (P <= 0.05) length, width, area, fresh and dry weight of clumps on multiplied plants. The R5 medium produced highest length (2.58 cm), width (2.16 cm), area (5.57 cm(2)), fresh weight (402.6 g) and dry weight (37.6 g) on regenerated clumps that showed healthier and green plant on culture medium. However, the regenerated clumps diameter was also significantly influenced (P <= 0.05) by the type and concentration of sugar (carbon sources) used. The highest clump diameter (7.27 cm) was obtained on R5 medium supplemented with 1.5% sucrose. Whereas, the high...
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