Cedrela odorata L. is a valuable tropical tree widely appreciated for its wood. This species confronts serious problems due to both overexploitation of its natural populations and its susceptibility to the Meliaceae borer Hypsipyla grandella, which destroys the apical meristems and produces structural deformations. The rapid introduction of new varieties through clonal forestry has been demonstrated to be the most effective way to improve the production of perennial plantation species. In this work, we report both a protocol for the rejuvenation of elite mature trees of C. odorata and the optimization of an in vitro culture system to scale up micropropagation. Several media formulations and the use of temporary immersion culture in bioreactors were evaluated. The addition of 20% coconut water to TY17 medium increased the number of adventitious shoots from hypocotyl segments to an average number of 4.68 shoots per explant. To replace coconut water and to define the culture medium, several cytokinins were tested at various concentrations; however, none of them produced the effect of coconut water. Rejuvenation of elite mature individuals was investigated by ex vitro grafting of mature tree twigs onto 3-mo-old juvenile trees.Although the grafting had a positive effect on the micropropagation of mature material, the multiplication rate of 1.5 new shoots per explant did not compare to the organogenic capacity of younger materials. Shoot and root elongation as well as acclimatization to ex vitro conditions were carried out in a temporary immersion culture of juvenile material using BioMINT® bioreactors. A 3.5-fold increase in shoot elongation and a 4-fold increase in root elongation were achieved compared to material cultured on semisolid media. Furthermore, this culture system allowed for 98% effectiveness in the soil adaptation of the in vitrogrown plants. The scaled-up multiplication capacity over a period of 6 mo calculated for the system is above 16,000 plants per mother plant with young materials but is only 125 with mature materials.
Agave tequilana Weber var. ‘Azul’ is grown for the production of tequila, inulin and syrup. Diverse bacteria inhabit plant tissues and play a crucial role for plant health and growth. In this study culturable endophytic bacteria were extracted from leaf bases of 100 healthy Agave tequilana plants. In plant tissue bacteria occurred at mean population densities of 3 million CFU/g of fresh plant tissue. Three hundred endophytic strains were isolated and 16s rDNA sequences grouped the bacteria into eight different taxa that shared high homology with other known sequences. Bacterial endophytes were identified as Acinectobacter sp., A. baumanii, A. bereziniae, Cronobacter sakazakii, Enterobacter hormaechei, Bacillus sp. Klebsiella oxytoca, Pseudomonas sp., Enterococcus casseliflavus, Leuconostoc mesenteroides subsp. mesenteroides and Gluconobacter oxydans. Isolates were confirmed to be plant growth promoting bacteria (PGPB) by their capacities for nitrogen fixation, auxin production, phosphate solubilization, or antagonism against Fusarium oxysporum AC132. E. casseliflavus JM47 and K. oxytoca JM26 secreted the highest concentrations of IAA. The endophyte Acinectobacter sp. JM58 exhibited the maximum values for nitrogen fixation and phosphate solubilization index (PSI). Inhibition of fungi was found in Pseudomonas sp. JM9p and K. oxytoca JM26. Bacterial endophytes show promise for use as bio-inoculants for agave cultivation. Use of endophytes to enhance cultivation of agave may be particularly important for plants produced by micropropagation techniques, where native endophytes may have been lost.
BackgroundMarine invertebrate-associated microbial communities are interesting examples of complex symbiotic systems and are a potential source of biotechnological products.ResultsIn this work, pyrosequencing-based assessment from bacterial community structures of sediments, two sponges, and one zoanthid collected in the Mexican Caribbean was performed. The results suggest that the bacterial diversity at the species level is higher in the sediments than in the animal samples. Analysis of bacterial communities’ structure showed that about two thirds of the bacterial diversity in all the samples belongs to the phyla Acidobacteria and Proteobacteria. The genus Acidobacterium appears to dominate the bacterial community in all the samples, reaching almost 80% in the sponge Hyrtios.ConclusionsOur evidence suggests that the sympatric location of these benthonic species may lead to common bacterial structure features among their bacterial communities. The results may serve as a first insight to formulate hypotheses that lead to more extensive studies of sessile marine organisms’ microbiomes from the Mexican Caribbean.Electronic supplementary materialThe online version of this article (doi:10.1186/0717-6287-47-67) contains supplementary material, which is available to authorized users.
Tabebuia donnell-smithii is a tropical tree species that is highly important as a forest crop due to both environmental and economic benefits. Wood from these trees is in high demand, achieving up to three times the price of wood from Pinus species. Commercial plantations of this species help to reduce the pressure on natural populations; however, plantations are few in number due to the lack of domesticated varieties and the relative unavailability of seeds. Therefore, there is an urgent need for a reliable source of plantlets suitable for commercial production. In the present work, we report the clonal propagation of T. donnell-smithii from twigs collected from elite trees. Emerging axillary shoots were used as a source of explants. Fungal contamination was a persistent problem but was partially overcome by brief exposure to low concentrations of chlorine. The stem fragments that were used as an explant source were cultured in woody plant medium containing 30 μM zeatin. This was the best condition for inducing adventitious shoot proliferation, producing 2.8 shoots per explant. Coconut water had an additional positive effect, increasing the number of shoots per explant by a factor of up to 3.6. The shoots were rooted in medium containing 20 μM indole-3-butyric acid. Ninety-four percent of the rooted plants survived the transfer to soil.
Spanish red cedar (Cedrela odorata L.) is a tropical timber tree native to the Americas from southern Mexico to northern Argentina. Commercial plantations are scarce and, consequently, natural populations are overexploited. Traditional propagation practices for the establishment of large-scale plantations have had limited success in this species due to the relative scarcity of seeds, its broad genetic diversity and the lack of domesticated varieties. In vitro clonal propagation provides an effective method to overcome this situation and increase the yield of commercial plantations through the rapid multiplication of elite materials. Somatic embryogenesis (SE) is one of the most promising strategies for tree propagation due to the possibility of producing artificial seeds, the ability to store and rapidly mobilize germplasm and the opportunity for genetic manipulation. We report here the induction of indirect SE in C. odorata from calli derived from immature zygotic embryos after 12 weeks of culture. Macroscopic, histological, and scanning electron microscopic analyses of the calli revealed the presence of embryogenic cell clusters that formed cotyledonary embryos with clear bipolar structures and no vascular connections with the mother tissue. Different media preparations containing combinations of diverse auxins and cytokinins are known to have different effects on the type and frequency of embryogenic structures. Embryo conversion was achieved using an MS-based medium [Murashige T, Skoog F (1962) Physiol Plant 15:473-497, 1962 supplemented with abscisic acid, and transfer to soil was successful at a rate of 75%. The method described here provides a basis for optimizing the clonal propagation and genetic manipulation of this valuable species.
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