To develop a system for Agrobacterium-mediated transformation of maize
(Zea mays L.), we have investigated histochemically the transient exp
ression of beta-glucuronidase (GUS) activity in maize seedling tissue
segments using binary vectors that allow minimal (pKIWI105 and pCNL1)
or undetectable (p35S-GUS-INT and pCNL56) levels of GUS activity in A.
tumefaciens. Tissue segments from three- to five-day-old sterile seed
lings of maize genotype A 188 were inoculated with A. tumefaciens. Fou
r days after inoculation, transient expression of GUS activity was fou
nd in mesocotyl segments originating from the intercalary meristem reg
ion. This GUS activity was specific to the vascular cylinder and was n
ot found in the internal cortical or epidermal layers, nor was it foun
d in mature mesocotyl tissue (segments 5 mm below the coleoptilar node
). Transient GUS activity was also detected in leaf and coleoptile tis
sues of shoot segments, but not in the shoot apex per se or in leaves
younger than the first leaf. Maize tissues inoculated with A. tumefaci
ens strains that harbour gusA-containing binary vectors but no Ti-plas
mid did not show GUS activity, supporting evidence from previous work
that vir gene activity was essential for the observed GUS activity. A.
tumefaciens strains containing different types of Ti-plasmids were al
so tested. A strain harbouring an agropine-type Ti-plasmid was the mos
t effective for expressing GUS activity in mesocotyl segments, whereas
a strain harboring a nopaline-type Ti-plasmid was most effective for
expression of GUS activity in the apical meristem-containing segment.
These results indicate that different interactions occurred between th
e different A. tumefaciens strains and the susceptible plant tissues.
Maize genotype specificity for GUS activity in mesocotyl tissues was o
bserved; variations in the cocultivation medium had a profound effect
on the frequency of expression of GUS activity.