Under in vitro conditions gametangial formation in bryophytes can be regulated by a variety of physical and chemical factors. Liverworts are relatively more sensitive to photoperiod than are mosses. With the exception ofRiccia crystallina all species of liverworts thus far investigated are either long-day or short-day plants. Among mosses, all butSphagnum plumulosum appear to be independent of photoperiod for the onset of the reproductive phase. The photoperiodic response in liverworts increases with increasing light intensity. Mosses, on the other hand, exhibit differential behavior.Leptobryum pyriforme andBartramidula bartramioides exhibit a linear relationship with light intensity for this response, whereasBryum argenteum, B. coronatum andBarbula gregaria have a specific light intensity requirement for optimal response. The photoperiodic effect in bryophytes is operative within certain temperature limits. Most bryophytes do not require a low temperature pretreatment for gametangial induction. Some become fertile in a broad temperature range, whereas others require a specific temperature. Among chemical factors, carbohydrates in general enhance gametangial formation.Riccia crystallina andBartramidula bartramioides develop gametangia in the presence of a carbohydrate, butBryum spp. (B. argenteum andB. coronatum) andBarbula gregaria respond even in its absence. High carbohydrate-nitrogen ratio seems more favorable for the onset of reproductive phase. Bryophytes respond differentially to various forms of nitrogen. Depletion of inorganic nitrogen (nitrate or ammonium) significantly favors gametangial induction. On the other hand, organic nitrogen (such as amino acids, peptone and urea) has differential effects on the enhancement of antheridial and archegonial formation in liverworts. Growth hormones have variable effects on gametangial induction. Indole-3-acetic acid increases archegonial formation inRiccia crystallina, but it is more favorable for antheridial production in mosses likeBryum coronatum, B. argenteum andBarbula gregaria. Gibberellins enhance antheridial formation in all the investigated bryophytes. Cytokinins stimulate induction of archegonia and inhibit antheridial formation inRiccia crystallina andBryum argenteum. Auxins, gibberellins and cytokinins also interact in eliciting the response. Iron and copper chelating agents such as EDTA and EDDHA favor vegetative growth and gametangial formation. InRiccia these chelates enhance archegonial production more than antheridial formation, but inBryum argenteum their effect is just the reverse. Salicylic acid, known to chelate iron and copper in certain animal systems, inhibits gametangial formation in most bryophytes, except inBartramidula in which it enhances vegetative growth as well as gametangial formation. InBryum argenteum the effect of these chelates is accompanied with marked changes in the endogenous levels of iron and copper. Iron appears to favor the onset of reproductive phase, whereas copper is inhibitory. Cyclic AMP, a well known mediator of hormone action in animal systems, enhances gametangial formation inBryum argenteum. The response is specific and is mimicked by phosphodiesterase inhibitors. Cyclic AMP also increases antheridial production inBryum coronatum andBarbula gregaria. InBryum argenteum it overcomes the inhibitory effect of ammonium ions and high concentrations of sucrose on gametangial formation. In addition to the above factors, pH and nutritional status of the medium also affect the onset of reproductive phase. Bryophytes exhibit maximum response in a definite pH range. Moreover, a specific change in pH of the medium is observed during gametangial initiation. Nutritional status, as affected by the concentration of nutrients in the medium, has varied effects on gametangial formation. In most instances dilution of the medium is more favorable for the response. Certain other factors, such as yeast extract and animal sex hormones, also enhance the formation of antheridia and archegonia. The onset of the reproductive phase involves metabolic changes in the differentiating tissues. Archegonial initiation is accomplished by intense metabolic activities, and in certain liverworts there is an increase in the content of carbohydrates, auxins, RNA and proteins, whereas the level of total nitrogen drops. Besides this a number of enzymes and phenolic compounds also exhibit marked qualitative and quantitative changes. Once gametangia are induced under in vitro conditions, fertilization can be accomplished by flooding the cultures bearing mature gametangia with sterile, distilled water. Development of sporophytes is markedly affected by temperature and nutritional status of the medium.