شماره ركورد كنفرانس :
5342
عنوان مقاله :
Effect of DTT and organic reagents on DNA quality and quantity of protein-rich seeds of chickpea (Cicer arietinum L.)
عنوان به زبان ديگر :
Effect of DTT and organic reagents on DNA quality and quantity of protein-rich seeds of chickpea (Cicer arietinum L.)
پديدآورندگان :
Ghaffari Akram a.ghaffari@areeo.ac.ir Registration and identification of plant varieties department, Seed and plant certification and registration institute (SPCRI), Agricultural Research, Education, and Extension Organization (AREEO), 31535-1516 Karaj, Iran , Jazayeri Mohammad Reza Registration and identification of plant varieties department, Seed and plant certification and registration institute (SPCRI), Agricultural Research, Education, and Extension Organization (AREEO), 31535-1516 Karaj, Iran , Oskouei Bita Seeds registration and certification department, Seed and plant certification and registration institute (SPCRI), Agricultural Research, Education, and Extension Organization (AREEO), 31535- 1516 Karaj, Iran
كليدواژه :
Chickpea , protein , DNA extraction , DTT , Organic reagent.
عنوان كنفرانس :
اولين همايش بين المللي و پنجمين همايش ملي علوم و تكنولوژي بذر ايران
چكيده فارسي :
Chickpea is a legume with richness in protein. Therefore, DNA isolation of seeds with proper purity is a great challenge for any molecular studies of this crop due to its abundance in protein and carbohydrates. In this research, a chickpea seeds collection from various seed classes of candidate varieties was tested for genetic purity. Three experiments were planned to find an appropriate protocol for DNA extraction of chickpea seed. SDS based seed extraction buffer used along with, two levels of DTT (Dithiothreitol) and PVP (Polyvinylpyrrolidone) including, 2% PVP with 0.4%, 0.8% DTT and applying organic reagents including phenol: chloroform (25:24, v/v) and chloroform: isoamyl alcohol (24:1, v/v). The data pointed out that applying DTT increase DNA yield that the highest DNA concentration achieved in 0.8% DTT 0, while increasing DTT to 0.8% had an adverse effect on DNA quality. Organic reagents along with 0.4% DTT presented the best result in terms of DNA quality. Both 260/280 and 260/230 ratios were above 2 and the DNA yield was acceptable for molecular studies. As a result, using DTT can increase DNA yield in chickpea seeds but not great impact on decreasing protein residuals. Exploiting organic reagents with 0.4% DTT was efficient to remove protein residual from chickpea seeds.
