Developmental and hormonal regulation of β-1,3-glucanase in tobacco

A highly sensitive and specific "rocket" immunoassay was used to measure the content of an endo-type β-1,3-glucanase (EC 3.2.1.39) in tissues of Nicotiana tabacum L. cv. Havana 425. We show that the accumulation of β-1,3-glucanase in cultured pith-parenchyma tissue is blocked by combinations of the auxin, α-naphthaleneacetic acid (NAA), and the cytokinin, kinetin. When tissues pre-incubated for 7 d on complete medium containing 2.0 mg·l(-1) NAA and 0.3 mg·l(-1) kinetin are transferred onto medium without hormones or with either hormone added separately, the β-1,3-glucanase content expressed per mg soluble protein increases approx. ten fold over a 7-d period. Under these inductive conditions, up to approx. 5% of the soluble protein is β-1,3-glucanase. The induction is inhibited by >90% when tissues are cultured over the same period on medium containing both hormones. This β-1,3-glucanase is developmentally regulated in the intact plant. It is a major component of the soluble protien in the lower leaves and roots but is not detectable in leaves near the top of the plant.