ABSTRACT
Abstract: It has learned that soybean is being affected by a floral disorder known as floral malady where plants fail to develop pod and do notattendfull maturity. For this floral disorder, we present a new methylation-sensitive amplified polymorphism (MSAP) approach for the evaluation of relative quantitative characteristics of non-methylation, hyper-methylation, hemi-methylation, and full methylation status of CCGG sequences, which are recognized by the isoschizomers HpaII and MspI. We applied a technique to analyze alterations in the cytosine methylation a popular Indian soybean (Glycine max L.) genotype, JS-335.The result revealed that in the symptomatic plant, out of 392 MSAP sites, 281 (71.68%), 33 (8.41%),38 (9.69%), and 40(10.20%) found to beun-methylated, hemi-methylated, fully methylated and hyper-methylated, respectively. Whereas, the MSAP profile of asymptomatic plants revealedout of 402MSAP sites, 330 (81.28%) was un-methylated, 22(5.41%) hemi-methylated,29(7.14%) fully methylatedand 25 (6.15%) hypermethylated. In comparison with asymptomatic(18.71%) plant, approximately 10% increased methylation was noted in symptomatic(28.31%) plantprofiles. The increased levels of methylation was recorded in the symptomatic plants about 28.31%and18.71% in asymptomatic. The study showed a higher epigenetic influence on JS-335 genotype of floral malady symptomatic than same genotype of asymptomatic plant. No pod formation in symptomatic plant induce genome wide changes either in promoter or coding region of gene(s) and DNA fragments showing polymorphism related to differences in pattern and extent of methylation associated with floral malady.
ABSTRACT
Shoot regeneration in safflower (Carthamus tinctorius 'AKS 207' and 'PKV Pink') genetically transformed using Agrobacterium was used for assessing various constraints to the efficiency of transformation including infection period, virulence induction medium, co-cultivation period, bacterial titre, selection regime, and the natural phenolic compound acetosyringone. Transformation frequency was promising with 8-10-day-old cotyledonary leaf explants. Therefore, explants of that age cultured on Agrobacterium minimal medium (AB) containing 100 µM acetosyringone were infected with Agrobacterium (cell titre 0.5 OD600nm) for 15 min followed by 48 h of co-cultivation on kanamycin-enriched medium (50 mg/L). Transformation of the shoots was confirmed using ß-glucuronidase (GUS) histochemical assay and polymerase chain reaction (PCR). With the transformation protocol thus optimized, the transformation frequency as determined using GUS assays was 54.0 % for AKS 207 and 47.6 % for PKV Pink. The corresponding figures using PCR were 27.0 and 33.3 %. The transformed shoots required 10-14 weeks of culture initiation but produced very few roots.
