LIFE HISTORY AND GROWTH OF LAURENCIA BRACHYCLADOS (RHODOPHYTA,CERAMIALES)1

Laurencia brachyclados Pilger from Hawai'i completed a “Polysiphonia-type” tri-phasic life history in 21 weeks in laboratory culture. Tetraspores developed into gametophytes in a nearly 1:1:1 ratio of females: males: non-reproductive. Carpospores were released as early as 21 days after mixing virgin female and male gametophytes. Cultured thalli showed a “guerilla type” growth form. Other Hawaiian Laurencia species in culture had longer maturation times or remained non-reproductive. Variation in life history schedules may influence Laurencia species coexistence and algal community structure.     

Plant regeneration from protoplasts of the monocotyledonous Haworthia magnifica v. Poelln.

Calli were initiated from flower buds, gynoecia and inflorescence segments of Haworthia magnifica v. Poelln. and subcultured on solid medium. Two liquid culture steps were necessary to prepare the calli for the isolation of protoplasts capable of sustained cell divisions. Plants were regenerated from protoplast-derived calli. The influence of both the osmolality of the culture media and exudates on the viability of protoplasts and protoplast-derived cell colonies is briefly discussed.     

In vitro reactions of sugarcane (Saccharum SP.) plantlets inoculated with 2 strains of Xanthomonas albilineans (Ashby) Dowson

The symptoms of the leaf scald disease can be reproduced in vitro through the inoculation of sugarcane tissue culture plantlets. The pathogen is detected in the inoculated plantlet and is maintained at the surface of the base of the plantlets grown in vitro. Two strains of X. albilineans belonging to different serovars and lysovars reacted like pathotypes. The importance of the plant incubation temperature is clearly demonstrated. Further, in vitro the disease goes through the same phase of latency as in the field.     

Somaclonal variation from Sorghum bicolor (L.) Moench cell culture

Plant Cell, Tissue and Organ Culture (PCTOC) - Sorghum bicolor (L.) Moench, plants were regenerated from 4 to 5 month old callus cultures originally derived from seedling explants. Somaclonal...     

In vitro propagation of Alstroemeria ‘Alsaan’

Plantlets were regenerated from Alstroemeria Alsaan rhizome tips cultured in vitro on solid and liquid media based on Murashige and Skoog salt formulation. The quality of the cultures was superior when intact rather than longitudinally sliced rhizome tips were used as explants and when a temperature of 8°C rather than 22°C was used at the initiation stage. More roots were produced on rhizome tips containing a rhizome apical meristem than on rhizome sections lacking such a meristem. Most (90%) of the rooted plantlets were successfully acclimatized and developed into true-to-type flowering plants.     

In vitro regeneration of apomictic bluestem grasses

Explants from immature inflorescences of four genotypes of Old World bluestem grasses, (Bothriochloa spp.), produced callus tissue on Linsmaier and Skoog (RM) and 1/2 Murashige and Skoog (1/2 MS) media containing high levels of growth regulators. Callus masses were composed of two distinct tissue types, one a compact, white, embryogenic portion (E calli), the other soft, translucent, gelatinous and nonembryogenic (NE calli). When transferred to medium with a reduced level of 2,4-D, and/or supplemented with zeatin, E callus underwent further organization culminating in shoot production. Light and scanning electron microscopy confirmed the embryogenic pathway of differentiation. Genotype significantly affected callus induction frequency and the number of plants regenerated. The RM medium induced more explants to initiate callus compared to the 1/2 MS medium. Age of the inflorescence explant, as indicated by size, was critical for callus induction. Inflorescences with racemes 8 mm in length were superior to older ones. Five-hundred-twenty-two plantlets were regenerated and grown to maturity.     

In vitro organogenesis and plant regeneration from leaves of Solanum candidum Lindl,S. quitoense Lam. (naranjilla) and S. sessiliflorum Dunal

Adventitious shoots and roots were regenerated from leaf segments of 3 Solanum species: S. candidum Lindl., S. quitoense Lam. and S. sessiliflorum Dunal. Leaf explants differentiated shoots on modified MS medium supplemented with 23–163 M kinetin and 0–5.7 µM indoleacetic acid (IAA). Excised shoots were induced to form roots by transfer to media with benzyladenine (BA) and naphthaleneacetic acid (NAA) at 0.09 and 0.11 µM respectively for S. quitoense and 0.01 µM NAA for S. candidum and S. sessiliflorum. Adventitious roots were produced directly from leaf explants with 0–140 µM kinetin and 0–5.7 µM IAA in combination. Rooted plants were successfully established in the greenhouse.     

Initiation and maintenance of long term somatic embryogenesis from anthers and ovaries of Vitis longii ‘Microsperma’

Anthers and ovaries of Vitis longii Microsperma produced embryogenic callus when cultured on solidified Murashige and Skoog medium with 5M 2,4-dichlorophenoxyacetic acid (2,4-D) and 1M benzyladenine (BA). The initial callus was short-lived. However, long-term embryogenesis from callus was maintained through serial transfers by careful selection of clustered embryos with subtending callus. Alternatively, long term culture maintenance was through secondary embryogenesis which occurred directly from previously formed embryos on medium lacking growth regulators. Somatic embryos were white, exhibited frequent pluricotyly and tended to be larger than zygotic embryos. Histology of embryogenic callus demonstrated the presence of lipid-like substances and abundant starch. Somatic embryos were attached to callus by narrow to wide suspensor-like structures and possessed typical epidermal, cortical, and vascular tissue. Embryo cells contained abundant lipid-like accumulations but no starch. Embryos germinated when placed on medium containing 1M BA and produced plants of normal appearance.     

In vitro proliferation of saffron (Crocus sativus L.) stigma

Excised young intact stigmas plus ovaries of Crocus sativus L. were cultured on Linsmaier-Skoog media supplemented with either a cytokinin or an auxin alone or in combinations. Benzyladenine and kinetin at concentrations of 0.1, 1, and 5 mgl^-1 supported growth, and crocin was biosynthesized in the stigmas in vitro. Auxins had little effect. Young excised single stigmas or half ovaries were also cultured so as to form stigma-like structures in order to explore a possible new approach to industrial production of the spice, saffron. On Linsmaier-Skoog and Nitsch media supplemented with kinetin at concentration of 1 or 5 mgl^-1 and alpha-naphthalene acetic acid or indole-butyric acid at concentration of 0.1 or 10 mgl^-1 in combinations, stigma-like structures appeared directly and indirectly (through meristematic tissue), grew and matured. The maximum number of structures were 75 per half ovary. Three kinds of yellow pigments including crocin were tentatively identified by TLC in the stigma-like structures as was the case for the in vivo grown natural stigma, although the contents were lower.Abbreviations BA benzyladenine - 2,4-D 2,4-dichlorophenoxy-acetic acid - IAA indole-acetic acid - IBA indole-butyric acid - NAA alpha-naphthalene-acetic acid - TLC thin layer chromatography