THE ROLE OF CELLULASE IN HORMONAL-REGULATION OF SHOOT MORPHOGENESIS IN TOBACCO CALLUS

A polyclonal antibody raised against cellulase (EC 3.2.1.4.) from callus of Nicotiana tabacum L. cv. Petit Havana SR1 reduced cellulase activity and induced shoot formation in tobacco callus in the presence of callus maintaining concentrations of auxin and cytokinin. Shoot induction as well as reduction of the cellulase activity was also obtained by withdrawing auxin from the callus medium. The effect of the two hormones on cellulase activity in the tobacco tissue was examined by varying the concentration of one of the hormones alpha-naphthylacetic acid (NAA) or benzylaminopurine (BAP) at a time while the other was kept at a level sufficient for either callus growth or shoot induction. While NAA stimulated the enzyme activity increasingly with concentration in the range 5 x 10(-7) M to 5 x 10(-5) M at both levels of BAP, BAP only stimulated the cellulase activity at an optimum concentration of 5 x 10(-6) M when NAA was present at a level sufficient to induce callus growth. The results point to a pivotal role of the downward regulation of cellulase in the initiation of shoot induction. A series of events leading to oriented cell divisions as a result of the lowered cellulase level during the initial phase of the morphogenetic process is discussed.