G. Hull et al., USE OF THE LACZ REPORTER GENE AS AN INTERNAL CONTROL FOR GUS ACTIVITYIN MICROPROJECTILE BOMBARDED PLANT-TISSUE, PLANT SCI, 120(2), 1996, pp. 153-160
A protocol is described for simultaneous histochemical detection of La
cZ and Gus activity in plant tissues after microprojectile bombardment
. The suitability of two different Gus substrates (Salmon-glcA, Magent
a-beta-D-glcA) is compared. This detection system is used to assay the
number of cells expressing either or both reporter gene. This techniq
ue is used as a qualitative assay to demonstrate the tissue specificit
y of a Hrgp promoter in maize embryos, and to measure ABA responsivene
ss of a Lea promoter in Arabidopsis. The promoter to be studied is lin
ked to the gus reporter gene and the lacZ reporter gene linked to the
CaMV 35S promoter is used as a constitutive internal control. The use
of an internal control drastically reduces the data variation inherent
to microprojectile bombardment.