Establishment of a micro-particle bombardment transformation system for Dunaliella salina.
In this study, we chronicle the establishment of a novel transformation system for the unicellular marine green alga, Dunaliella salina. We introduced the CaMV35S promoter-GUS construct into D. salina with a PDS1000/He micro-particle bombardment system. Forty eight h after transformation, via histochemical staining, we observed the transient expression of GUS in D. salina cells which had been bombarded under rupture-disc pressures of 450 psi and 900 psi. We observed no GUS activity in either the negative or the blank controls. Our findings indicated that the micro-particle bombardment method constituted a feasible approach to the genetic transformation of D. salina. We also conducted tests of the cells' sensitivity to seven antibiotics and one herbicide, and our results suggested that 20 microg/ml of Basta could inhibit cell growth completely. The bar gene, which encodes for phosphinothricin acetyltransferase and confers herbicide tolerance, was introduced into the cells via the above established method. The results of PCR and PCR-Southern blot analyses indicated that the gene was successfully integrated into the genome of the transformants.
Tan
Congping
C
Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, P.R.China.
Qin
Song
S
Zhang
Qun
Q
Jiang
Peng
P
Zhao
Fangqing
F
D000613:Aminobutyrates / Q000494:pharmacology; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D019470:Biolistics; D019149:Bioreactors; D015139:Blotting, Southern; D000460:Chlorophyta / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development* / Q000378:metabolism; D018744:DNA, Plant / Q000737:chemistry / Q000235:genetics; D005966:Glucuronidase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008826:Microbial Sensitivity Tests; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D014170:Transformation, Genetic / Q000502:physiology*
D000613:Aminobutyrates; D000900:Anti-Bacterial Agents; D018744:DNA, Plant; C003121:phosphinothricin; D005966:Glucuronidase