Phenocritical times in the process of in vitro shoot organogenesis.

Shoot organogenesis occurs when leaf explants of Convolvulus arvensis are cultured on Murashige and Skoog salts, sucrose, vitamins, and 0.05 mg/liter IAA with 7.0 mg/liter 2-isopentenyl adenine. Under the influence of this shoot inducing medium (SIM), the explants become competent for the organogenic effects of SIM and eventually become determined for shoot formation. The induction process includes five separate transient sensitivities to inhibitors. Such stage-specific inhibitions reflect phenocritical times in development rather than general metabolic toxicities. The phenocopying agents are tri-iodobenzoic acid (TIBA), sorbitol, ribose, ammonium ion, and acetylsalicylic acid. The process of in vitro shoot organogenesis from leaf explants is now seen to include a series of discrete steps which precede morphological differentiation. An initial dedifferentiation process results in the formation of competent callus tissue along the cut edges of the explant. Under the influence of the phytohormone balance in SIM, shoot organogenic induction proceeds. This process involves a time which is sensitive to inhibition by salicylates followed by a time sensitive to TIBA which is followed in turn by a time sensitive to sorbitol and culminates in cells or groups of cells determined for shoot formation. This process also includes a time sensitive to inhibition by ribose, although its place in the order of events is not yet firmly assigned. There is also a sensitivity to ammonium ion (or lack of nitrate) at or near the time the explant becomes determined for shoot production.

A morphogenetically competent soybean suspension culture.

A morphogenetically competent suspension culture was derived from embryonic axes of Glycine max cv. Mitchell. The cultural history included visual selection for nonfriable, embryo-like structures, recurrent selection in a regime of 2,4-dichlorophenoxyacetic acid exposure and withdrawal, and the replacement of the nitrogen in a Murashige and Skoog salts-based medium with 20 millimolar ammonium citrate. The embryoids produced by this suspension are capable of completing plantlet development. The suspension can be maintained by serial subculture.

Competence and determination in the process of in vitro shoot organogenesis.

Leaf explants of Convolvulus arvensis produce shoots when cultured on Murashige and Skoog salts, sucrose, vitamins and 0.05 mg/liter IAA plus 7.0 mg/liter 2-isopentenyl adenine. Shoot-inducing, root-inducing, or callus-inducing medium (SIM, RIM, or CIM) will cause small amounts of callus to form at the cut edges of the explant. This first-formed callus is developmentally interchangeable: SIM induces shoots in callus formed on CIM or SIM with equal effect and efficiency. Once induction begins in competent callus, the callus is no longer interchangeable. Under the continued influence of SIM, cells, or groups of cells become determined for shoot formation. This determination is strongly canalized for shoot formation: subsequent transfer to root-inducing medium does not affect the formation of shoots by the explant. The control of organogenesis by the auxin/cytokinin balance must occur between the time the tissue becomes competent and the time it is determined for shoot (or root) development. It is not known whether this control is a single or multiple phenomenon.