A rapid and reproducible method for isolating genomic DNA from a few crop plants suitable for polymerase chain reaction-based genotyping
Journal Title: Journal of Applied and Natural Science - Year 2017, Vol 9, Issue 2
Abstract
As most of the molecular markers in crop molecular breeding programmes are successful based on polymerase chain reaction (PCR), the isolated genomic DNA must be suitable for the same. Though PCR is a robust method and in most of the cases requires only a minute amount of genomic DNA as template, removal of potential PCR-inhibitory factors is quite important. The present work reports the optimization of a rapid genomic DNA isolation method, suitable for PCR-based genotyping of plants. As very minute amount of the genomic DNA isolated in this rapid method was found to be sufficient for PCR, a researcher is capable to go for several hundred independent PCR from single isolation. The method was validated in 4 different crops (wheat, tomato, brinjal and cauliflower) using different PCR-based molecular markers. In case of wheat, genomic DNA isolated in this method was found to be suitable PCR using the specific marker for the detection of the Lr34 gene. For tomato, genomic DNA isolated in this method was successfully used with the molecular markers for the detection of resistance alleles for yellow leaf curl disease and root knot disease. In case of brinjal, the isolated genomic DNA was found to be suitable for simple sequence repeat (SSR) marker assay. In a similar way, genomic DNA isolated in this method from cauliflower leaves was observed to be suitable for amplifying a gene of ~1.5 kb length. Thus, this method will be quite helpful to expedite marker assisted selection of plants in plant molecular breeding programmes.
Authors and Affiliations
Vikash Kumar, Pawan Kumar, Tirthartha Chattopadhyay
A study on geospatial technology for detecting and mapping of Solenopsis mealybug (Hemiptera: Pseudococcidae) in cotton crop
Detection of crop stress is one of the major applications of remote sensing in agriculture. Many researchers have confirmed the ability of remote sensing techniques for detection of pest/disease on cotton. The objective...
Quality characteristics of white cabbage juice stored using thermal processing and addition of chemical additives
Cabbage (Brassica oleracea L. var. capitata) is one of the most important vegetables grown worldwide. Cabbage is valued for medicinal purposes in treating headaches, gout, diarrhea and peptic ulcers. The beneficial phyto...
Assessment of nutrient ratios in the leaf of sugarcane grown in Theni district of Tamil Nadu, India
To assess the impact of nutrient ratios N/P, N/K and N/S on sugarcane productivity, a survey was conducted during its grand growth stage (October, November and December) influenced by various yield levels based on soil q...
Seed size and its influence on germination, seedling growth and biomass in Saraca asoca (Roxb). De Wilde, critically endangered tree species of Western ghats, India
Saraca asoca (Roxb). De Wilde is one of the high traded medicinal plant species of India. There is a huge demand for bark of this species both in domestic and international markets. There is a demand for quality planting...
Biosorption technology for removal of metallic pollutants-An overview
The main sources of metallic pollutants to the environment are the diffuse sources such as forests and agricultural soils as well as industrial and municipal wastes, which are either discharged directly or transported in...