Effects of auxin and light treatments of donor plants on shoot production fromindica-type rice (Oryza sativa L.)

An efficient clonal propagation procedure for a Brazilianindica rice subspecies was developed with shoot apex explants. Shoot apices were excised from 4-d-old seedlings and cultured on MS medium supplemented with 8.9 μM 6-benzyladenine. The efficiency of shoot production was influenced by growth regulators and light treatments to the donor plant. Explants derived from seedlings growth in the presence of 10.7 μM naphthaleneacetic acid and in the absence of light showed significantly increased regeneration capacity as compared to control explants. Anatomical analysis of the new shoot meristems revealed that they originated from preexisting apical and axillary meristem as well as from the mesocotyl parenchyma.     

Proliferation of meristematic clusters in disposable presterilized plastic bioreactors for the large-scale micropropagation of plants

Summary Proliferation of meristematic clusters of several plants in an inexpensive airlift bioreactor system, consisting of a disposable presterilized light transmittable plastic film vessel is described. The optimal shape, size, and structural function of the disposable plastic bioreactor are based on the bubble column and airlift glass bioreactors. The disposable bioreactors are designed in a conical configuration with a single inoculation and harvest port and multiple use dispensing and mixing accessories. Shearing damage and foaming problems known to exist in bioreactors due to the plant's rigid cell wall and size were greatly reduced in the disposable plastic bioreactors. The disposable bioreactors were used for propagule proliferation and growth, using meristem and bud clusters of potato, fern, banana, and gladiolus. The clusters' biomass increased five-to eightfold over a period of 26–30 d, depending on the species. The clusters were separated mechanically by a chopper made of a grid of knives. The chopped propagules were inoculated to agar medium for further growth and developed into transplantable plants. In the case of gladiolus and potato, corms and tubers developed in a sucrose-elevated storage organ induction medium, respectively, after the initial formation of small shoots. The plantlets and storage organs were transplanted to an acclimation greenhouse and continued to grow with a 95–100% survival, depending on the species. Plant development was followed for a period of 16 wk in fern and 12–14 wk in potato, banana, and gladiolus and normal shoot and leaf growth was observed. The feasibility of large-scale liquid cultures for plant micropropagation is discussed.     

Micropropagation of licorice (Glycyrrhiza glabra L.) through shoot tip and nodal cultures

Summary Apical and axillary buds ofGlycyrrhiza glabra commonly known as licorice, a plant of repute in the Indian system of medicine, were used for induction of adventitious shoots. For induction of multiple shoots, Murashige and Skoog’s (MS) medium with N⁶-benzyladenine (BA, 0.88–8.87 μM) was used. Reduction in major salts of MS medium enhanced the multiplication ratio up to 1∶10. Plants transferred to the greenhouse showed 90% survival. The present work describes a stepwise protocol for production ofGlycyrrhiza glabra plants on simple minimal media, where very high multiplication rates with healthy root systems were obtained. Roots being the organ of commercial importance, the protocol has tremendous potential.     

Somatic embryogenesis and plant regeneration from immature zygotic embryos of Japanese larch (Larix leptolepis)

Embryogenic tissue was initiated using LM, LP and MS media from open-pollinated immature embryos of Larix leptolepis. The initiation frequency varied with collection dates. The highest frequencies of embryogenic tissue initiation (60, 67 and 59% on LM, LP and MS media, respectively) were observed from cones collected on July 30. At this time, all the excised embryos were at the cotyledonary stage. ABA over a wide concentration and length of exposure range did not promote maturation, but was beneficial in reducing precocious germination. Of over 400 plants regenerated, 72 were transplanted into soil mixtures and to date, 69 of these (95%) have survived. This revised version was published online in June 2006 with corrections to the Cover Date.     

Enhancement of in vitro growth of papaya multishoots by aeration

Efficient micropropagation of papaya (Carica papaya L.) has become crucial for multiplication of specific sex types of papaya or transgenic lines resistant to virus infection. In this study, aeration at different intervals with a 0.02 μm filter disc in the closure of culture flasks ensured exchange of gas components. The effect of aeration on development of multibuds to multishoots was investigated. Multibuds grown in culture flasks after one-week without aeration followed by a two-week aeration treatment caused a 41% increase in the number of shoots ≥0.5 cm, 42% increase in leaf expansion, and 17% increase in leaf numbers in comparison with unaerated materials. Ethylene and oxygen concentrations in the culture flasks were measured by gas chromatography and oxygen electrode at weekly intervals during the culture period. Oxygen concentrations were slightly different between aerated and unaerated culture flasks. Ethylene in the unaerated flask reached the highest level (0.11 ppm) 2 weeks after the treatment, while accumulation of ethylene in the aerated flasks was not detected. The multishoots grown for 3 weeks without aeration showed growth retardation on leaves and epinasty on petioles. This revised version was published online in June 2006 with corrections to the Cover Date.     

In vitro regeneration from leaf explants of Neoregelia cruenta (R. Graham) L.B. Smith,an endemic bromeliad from Eastern Brazil

An efficient plant regeneration system was developed for the induction of direct shoot formation from leaves derived from seedlings of Neoregelia cruenta, an endemic Bromeliaceae of Eastern Brazil. Shoot differentiation occurred directly from the leaf bases. In vitro responses were influenced by seedling age and growth regulator combinations. Highest regeneration rates were obtained from explants excised from 7-week-old seedlings cultured in the presence of 22 μM BA and 2.5 μM NAA. Shoot conversion to whole plants was most effective in shoots formed in response to 4.4 or 8.8 μM BA combined with 2.5 μM NAA. This revised version was published online in June 2006 with corrections to the Cover Date.     

In vitro shoot culture and microtuber induction in the steroid yam Dioscorea composita Hemsl

The individual effects of sucrose, plant growth regulators and basal salt media formulations were investigated on microtuber induction and development in shoot cultures of the steroid yam Dioscorea composita. Sucrose at 8% (w/v) was the single most significant medium constituent for microtuber induction. Of the four cytokinins tested, 6-benzyladenine at 1.25 and 2.5 μM showed strong inhibitory effects on microtuber induction. By contrast, the auxins α-naphthaleneacetic acid and indole-3-butyric acid at 5.0 μM showed striking promotive effects on microtuber induction and growth. In the presence of either one of these auxins at 5.0 μM shoot cultures produced microtubers weighing 300–400 mg fresh weight whilst kinetin, 6-(γ,γ-dimethylallylamino)-purine, 6-benzyladenine and abscisic acid failed to promote microtuber growth (microtubers weighed generally <200 mg). Media formulations Lloyd and MacCown and White supported the lowest frequencies of microtuber induction when kinetin was present at 2.5 μM. Anderson Rhododendron was as effective as Murashige and Skoog overall in promoting both microtuber induction and growth. When removed from cultures and planted in sterilized moist sand, microtubers sprouted readily (60–87% within 2 weeks) and produced vigorous shoot growth and after 5–7 months minitubers of sizes (30–80 g) suitable for direct field planting. This revised version was published online in June 2006 with corrections to the Cover Date.     

Miscanthus×giganteus plant regeneration: effect of callus types,ages and culture methods on regeneration competence

The perennial rhizomatous grass, Miscanthus×giganteus is an ideal biomass crop due to its rapid vegetative growth and high biomass yield potential. As a naturally occurring sterile hybrid, M. ×giganteus must be propagated vegetatively by mechanically divided rhizomes or from micropropagated plantlets. Plant regeneration through somatic embryogenesis is a viable approach to achieve large‐scale production of plantlets in tissue culture. Effect of the callus types, ages and culture methods on the regeneration competence was studied to improve regeneration efficiency and shorten the period of tissue culture in M. ×giganteus. Shoot‐forming calli having a yellow or white compact callus with light‐green shoot‐like structures showed the highest regeneration frequency. Percentage of shoot‐forming callus induction from immature inflorescence explants was 41% on callus induction medium containing 13.6 μM 2,4‐d and 0.44 μM benzyladenine (BA). The use of a regeneration medium containing 1.3 μM NAA and 22 μM BA was effective at shortening the incubation period required for plantlet regeneration, with 69% of total regenerated plantlets obtained within 1 month of incubation on regeneration medium. Embryogenic‐like callus morphotype could maintain regeneration competency for up to 1 year as suspension cultures. Field grown regenerated plants showed normal phenotypic development with DNA content and plant heights comparable to rhizome propagated plants. Winter survival rates of the regenerated plants planted in 2006 and 2007 at the University of Illinois South Farm, Urbana‐Champaign, Illinois, were 78% and 56%, respectively.     

Somatic embryogenesis from mature <Emphasis Type="Italic">Bambusa balcooa</Emphasis> Roxburgh as basis for mass production of elite forestry bamboos

A reliable protocol for mass propagation via somatic embryogenesis in mature bamboos has been established using pseudospikelets of Bambusa balcooa. Fourty percent of the explants gave rise to multiple regenerants within 4 months. This conversion rate is sufficiently high to use the process in commercial mass production. Further, shoot apical meristems can also be used as primary explants without lost of efficiency. Regenerated plants were uniform and identical to the mother plant and to plants obtained by axillary branching with respect to growth characteristics and morphology. Furthermore, epigenetic changes could not be detected by Methylation Sensitive AFLP (MSAP). During the complete process no changes in ploidy level could be observed. The process allows for a cost reduction for this tropical bamboo for forestry of up to 57% compared to micropropagation via axillary branching. For the first time, a reliable process based on somatic embryogenesis has been developed that is well suited for commercial micropropagation of elite mature bamboos.