Cloning of Candidate Genes from Seashore Paspallum

Benjamin Brickle

Abstract

Drought and salt stresses limit the amount of agricultural production every year. In this study, five predicted genes (m11, m23, m26, m28, and m31) that responded to both stresses were chosen from RNAseq database of seashore paspallum (Paspallum vaginatum), which is a warm-seasoned turf grass that is native to coastal regions. This grass is tolerant to many environmental factors such as salt and drought. The genes were cloned, and their effectiveness was evaluated in the model species Arabidopis via Agrobacterium-mediated plant transformation. In the research, rapid amplification of cDNA ends, DNA digestion, DNA ligation, DNA transformation into E. coli and Agrobacterium, colony PCR to detect the gene harboring in the host cell, plasmid DNA isolation, and gel electrophoresis techniques were used to conduct the experiments. Five and three prime ends of one of the candidate genes, m11, was cloned and sequenced. With these results, full length cDNAs can be cloned based on this sequence. In the future, the gene will be delivered into Agrobacterium via electroporation and transformed into Arabidopis using the floral dip method. The transgenic plants will be tested to see if the candidate gene positively impacts the plant’s response to salt and drought stresses. This strategy can also be implemented to crop species, which would lead to more resistant crops. The result would be an increase in agricultural production every year.

 
Mar 30th, 9:45 AM

Cloning of Candidate Genes from Seashore Paspallum

Founders Hall 114 A

Drought and salt stresses limit the amount of agricultural production every year. In this study, five predicted genes (m11, m23, m26, m28, and m31) that responded to both stresses were chosen from RNAseq database of seashore paspallum (Paspallum vaginatum), which is a warm-seasoned turf grass that is native to coastal regions. This grass is tolerant to many environmental factors such as salt and drought. The genes were cloned, and their effectiveness was evaluated in the model species Arabidopis via Agrobacterium-mediated plant transformation. In the research, rapid amplification of cDNA ends, DNA digestion, DNA ligation, DNA transformation into E. coli and Agrobacterium, colony PCR to detect the gene harboring in the host cell, plasmid DNA isolation, and gel electrophoresis techniques were used to conduct the experiments. Five and three prime ends of one of the candidate genes, m11, was cloned and sequenced. With these results, full length cDNAs can be cloned based on this sequence. In the future, the gene will be delivered into Agrobacterium via electroporation and transformed into Arabidopis using the floral dip method. The transgenic plants will be tested to see if the candidate gene positively impacts the plant’s response to salt and drought stresses. This strategy can also be implemented to crop species, which would lead to more resistant crops. The result would be an increase in agricultural production every year.