Pierce's Disease
Research Updates

piercesdisease.cdfa.ca.gov

What is Pierce's Disease?

Pierce's Disease is a bacterial infection, which is spread by bugs that feed on grapevines, particularly the "glassy winged sharpshooter." Grapevines that become infected with PD can quickly become sick and die.

glassy-winged sharpshooter

High-efficiency biolistic co-transformation and regeneration of Chardonnay (Vitis vinifera L.) containing npt-II and antimicrobial peptide genes


  • Author(s): Kikkert, Julie; Reisch, Bruce; Vidal, J.; Wallace, P.;
  • Abstract: A reliable and efficient system for transformation and regeneration of Chardonnay (Vitis vinifera L.) plants via microprojectile bombardment was developed. Improvements over the previous biolistic transformation system included: (1) the use of gold particles for bombardment; (2) step-wise selection at 10 then 15 mg/l kanamycin; and (3) embryo induction at 27degreesC. Embryogenic cell cultures were either bombarded with pBI426, which contains the reporter gene gus (uidA) coding for beta-glucuronidase (GUS), or were co-bombarded with pSAN237 carrying the npt-II (neomycin phosphotransferase II) selectable marker gene, and a second plasmid with an antimicrobial peptide gene. A large number of transient (7,883+/-1,928) and stable (46+/-32) blue spots per plate at 2 and 95 days after bombardment, respectively, were obtained according to GUS expression analyses. A total of 447 putative transgenic embryos was harvested from 84 bombarded plates. From these embryos, 242 (54%) were regenerated into plants within the first year of the experiment. Southern blot analyses confirmed integration of the transgenes into the grape genome. Co-transformation was tested with four separate antimicrobial constructs. The co-transformation frequency of unlinked genes was 48% as measured by polymerase chain reaction (PCR), and 56% as estimated by dot blot hybridization. Expression of the gus gene, and PCR and Southern blot analyses of npt-II and antimicrobial genes from regenerated plants document stable transformation of Chardonnay and establish the parameters for highly-efficient biolistic transformation in V. vinifera.
  • Publication Date: Nov 2003
  • Journal: Plant Cell Reports