In Vitro Culture Of Five Moss Species

Brachymenium nepalense Hook., Macromitrium cavaleriei Card.& Thér., Macrocoma sullivantii(Müll. Hal.) Grout., Bryum pallescens Schleich.ex Schwaegr. var. pallescensare and Leucobryum glaucum(Hedw.) A?ngstr. Are fivefour ornamental moss species as micro-landscape and vertical greening materials. In the present paper, we conducted their culture in vitro to study the influences of nutrient medium types, element concentration, sugar types, sucrose concentration, p H, hormone at different concentrations on the protonemal and gameophytic development and growth.The main results are as follows:(1) The optimal media for Brachymenium nepalense to grow in vitro are modified knop’s medium with NAA(0.5mg.L-1), 6-BA(0.05 mg.L-1), sucrose(10g.L-1), agar(6.0 g.L-1) and p H 6. For Brachymenium nepalense to grow and reprocue, their explants were incubated at a constant temperature of 23 ± 2℃ and a light intensity of 2000 lx with a 14-h light / 10-h dark regime. The modified Knop ’s medium is more suitable than MS medium for Brachymenium nepalenseto grow. The growth and differentiation of the protonemae of Brachymenium nepalense were not only related with medium types, but also with the medium concentration. Modified knop’s medium with KH2PO4 at 125 mg.L-1 is benefit for Brachymenium nepalense to grow, and reducing phosphorus content will inhibit the growth of the protonemae and gametophytes of B. nepalense. Adding Calcium,Glucose and sucrose into modified knop’s medium at a suitable concentration could promote the growth of B. nepalense, while fructose would inhibit the growth of B. nepalens. The medium at weak acidic condition is more suitable for the growth of B. nepalense gametophytes, and acidic medium is suitable for the growth of its protonemae. Among the factors including medium types, medium concentration, sucrose concentration, p H, nitrogen, calcium, phosphorus, different sugars and hormones, hormones have relatively distinct effect on the growth of B. nepalensemost. Adding NAA and 6-BA at suitable concentration could promote the growth of the moss species.(2) In vitro, Macromitrium cavaleriei was able to grow well in 1/3 MS withsucrose(6g.L-1) and agar(6g.L-1)) completely at a constant23 ± 2℃ and a light intensity of 2000lx;The gametophytes of Macromitrium cavaleriei grow better in MS medium than in modified knop’s medium. The MS medium with macro-element compounds at relatively low concentration is good for its gametophytes to grow. Adding NAA at low concentration into MS medium could promote the growth of moss gametophyte, while cytokinin 6-BA and NAA at high concentration would severely inhibite the growth of M. cavalerfiei gametophytes.(3)The gametophytes of Macrocoma sullivantii grew well in vitro on the media with KNO3(125mg.L-1), sucrose(10g.L-1), agar(6.0 g.L-1) and p H6, incubated at constant temperature of 23 ± 2℃ and a light intensity of 2000 lx with a 14-h light / 10-h dark regime. Modified knop ’s medium is better than MS medium for Macrocoma sullivantiito to grow, which is consistent with the result of Brachymenium nepalense. If the concentrations of medium, macro-element compounds and sucrose are too high or too low, the gametophytes of Macrocoma sullivantiig were not able to row well, they only grew well on the media at an appropriate concentration. The tested hormones in the present paper showed their inhibiting effects on the growth of the moss gametophyte. On the modified knop’s media with appropriate nitrogen content, the gametophytes differentiated and grew well.(4) For the gametophytes of Bryum pallescens, their optimal cultural conditions included a modified knop ’s medium with agar(6g.L-1), sucrose(10g.L-1), and reducing KNO3(125mg.L-1), at p H6. The explants were suggested to be cultured at a constant temperature of 23 ± 2℃ and a light intensity of 2000 lxwith a 14-h light / 10-h dark regime. The modified knop ’s medium is better than MS medium for Bryum pallescens gametophyte’s growth. The concentration of the media had not district effects on the growth of the gametophytes of B. pallescens. The other factors, such as the contents of macro-element and compounds, Ca2+ and P also didn’t have distinct influences on the growth of the gametophytes of Bryum pallescens. The modified knop’s, if with macroelements and compounds at low concentration, or with sucrose at low concentration, or even without sucrose at all, or were diluted by 20 times, the gametophytes of Bryum pallescens would grow well. Adding NAA or 6-BA into the modified knop ’s medium would inhibit the growth of the moss gametophytes, while adding NAA would promote the formation of the rhizoids of B. pallescens. The gametophytes of B. pallescens on the modified knop’s medium with nitrogen at half concentration of that of the normal one grew well.(5)The gametophytes of Leucobryum glaucum produced by in vitro tissue culture were inoculated on a matrix with peat, vermiculite and perlite at different proportions, and adding knop’s nutrition liquid at different concentrations. After a 60-day cultivation, the chlorophyll content, length, total fresh weight and branch number of the gametophytes of L. glaucum were measured. The results showed that L. glaucum could grow well on the medium with a proportion of peat, vermiculite and perlite at 2:2:1, together with 1/5 Knop’s nutrition liquid.Based on the above work, we proposed two invention patents in relation to methods to fast producing gametophytic of Brachymenium nepalenseand Brachymenium nepalense in vitro.How to quantatively report gameophytic growth indices of moss in vitro has notbe soloved yet. In the present paper, we proposed to apply a Win Rhizo root systemanalyzier to determine the gameophytic morphological indices by using theirscanned photos, which could be consulted in the similar work in the future.