CLASS-I BETA-1,3-GLUCANASES IN THE ENDOSPERM OF TOBACCO DURING GERMINATION

Rupture of the seed coat and rupture of the endosperm are separate events in the germination of Nicotiana tabacum L. cv Havana 425 seeds. Treatment with 10(-5) M abscisic acid (ABA) did not appreciably affect seed-coat rupture but greatly delayed subsequent endosperm rupture by more than 100 h and resulted in the formation of a novel structure consisting of the enlarging radicle with a sheath of greatly elongated endosperm tissue. Therefore, ABA appears to act primarily by delaying endosperm rupture and radicle emergence. Measurements of beta-1,3-glucanase activity, antigen content, and mRNA accumulation together with reporter gene experiments showed that induction of class I beta-1,3-glucanase genes begins just prior to the onset of endosperm rupture but after the completion of seed-coat rupture. This induction was localized exclusively in the micropylar region of the endosperm, where the radicle will penetrate. ABA treatment markedly inhibited the rate of beta-1,3-glucanase accumulation but did not delay the onset of induction. Independent of the ABA concentration used, onset of endosperm rupture was correlated with the same beta-1,3-glucanase content/seed. These results suggest that ABA-sensitive class I beta-1,3-glucanases promote radicle penetration of the endosperm, which is a key limiting step in tobacco seed germination.