Impact of two arbuscular mycorrhizal fungi on Arundo donax L. response to salt stress.

MAIN CONCLUSION: AM symbiosis did not strongly affect Arundo donax performances under salt stress, although differences in the plants inoculated with two different fungi were recorded. The mechanisms at the basis of the improved tolerance to abiotic stresses by arbuscular mycorrhizal (AM) fungi have been investigated mainly focusing on food crops. In this work, the potential impact of AM symbiosis on the performance of a bioenergy crop, Arundo donax, under saline conditions was considered. Specifically, we tried to understand whether AM symbiosis helps this fast-growing plant, often widespread in marginal soils, withstand salt. A combined approach, involving eco-physiological, morphometric and biochemical measurements, was used and the effects of two different AM fungal species (Funneliformis mosseae and Rhizophagus irregularis) were compared. Results indicate that potted A. donax plants do not suffer permanent damage induced by salt stress, but photosynthesis and growth are considerably reduced. Since A. donax is a high-yield biomass crop, reduction of biomass might be a serious agronomical problem in saline conditions. At least under the presently experienced growth conditions, and plant-AM combinations, the negative effect of salt on plant performance was not rescued by AM fungal colonization. However, some changes in plant metabolisms were observed following AM-inoculation, including a significant increase in proline accumulation and a trend toward higher isoprene emission and higher H2O2, especially in plants colonized by R. irregularis. This suggests that AM fungal symbiosis influences plant metabolism, and plant-AM fungus combination is an important factor for improving plant performance and productivity, in presence or absence of stress conditions.

Cryotechniques for the Long-Term Conservation of Embryogenic Cultures from Woody Plants.

Since its development in the 1960s, plant cryopreservation is considered an extraordinary method of safe long-term conservation of biological material, as it does not induce genetic alterations and preserve the regeneration potential of the stored material. It is based on the storage of explants at cryogenic temperatures, such as the one of liquid nitrogen (-196 °C), where the metabolism within the cells is suspended; thus, the time for these cells is theoretically "stopped". Cryopreservation is particularly important for embryogenic cultures, as they require periodic subculturing for their maintenance, and this, in turn, increases the risk of losing the material, as well as its embryogenic potential. Periodic re-initiation of embryogenic cultures is possible; however, it is labor intensive, expensive, and particularly difficult when working with species for which embryogenic explants are available only during a limited period of the year. Among various methods of cryopreservation available for embryogenic cultures, slow cooling is still the most common approach, especially in callus cultures from softwood species. This chapter briefly reviews the cryopreservation of embryogenic cultures in conifers and broadleaf trees, and describes as well a complete protocol of embryogenic callus cryopreservation from common ash tree (Fraxinus excelsior L.) by slow cooling.

Potential applications of cryogenic technologies to plant genetic improvement and pathogen eradication.

Rapid increases in human populations provide a great challenge to ensure that adequate quantities of food are available. Sustainable development of agricultural production by breeding more productive cultivars and by increasing the productive potential of existing cultivars can help meet this demand. The present paper provides information on the potential uses of cryogenic techniques in ensuring food security, including: (1) long-term conservation of a diverse germplasm and successful establishment of cryo-banks; (2) maintenance of the regenerative ability of embryogenic tissues that are frequently the target for genetic transformation; (3) enhancement of genetic transformation and plant regeneration of transformed cells, and safe, long-term conservation for transgenic materials; (4) production and maintenance of viable protoplasts for transformation and somatic hybridization; and (5) efficient production of pathogen-free plants. These roles demonstrate that cryogenic technologies offer opportunities to ensure food security.

In vitro propagation of olive (Olea europaea L.) by nodal segmentation of elongated shoots.

Olive (Olea europaea L.), long-living, ever-green fruit tree of the Old World, has been part of a traditional landscape in the Mediterranean area for centuries. Both the fruits consumed after processing and the oil extracted from the fruits are among the main components of the Mediterranean diet, widely used for salads and cooking, as well as for preserving other food. Documentations show that the ancient use of this beautiful tree also includes lamp fuel production, wool treatment, soap production, medicine, and cosmetics. However, unlike the majority of the fruit species, olive propagation is still a laborious practice. As regards traditional propagation, rooting of cuttings and grafting stem segments onto rootstocks are possible, former being achieved only when the cuttings are collected in specific periods (spring or beginning of autumn), and latter only when skilled grafters are available. In both the cases, performance of the cultivars varies considerably. The regeneration of whole plants from ovules, on the other hand, is used only occasionally. Micropropagation of olive is not easy mainly due to explant oxidation, difficulties in explant disinfection, and labor-oriented establishment of in vitro shoot cultures. However today, the progress in micropropagation technology has made available the complete protocols for several Mediterranean cultivars. This chapter describes a micropropagation protocol based on the segmentation of nodal segments obtained from elongated shoots.

In vitro propagation of peanut (Arachis hypogaea L.) by shoot tip culture.

Peanut (Arachis hypogaea L.), also known as groundnut, is the most important species of Arachis genus, originating from Brazil and Peru. Peanut seeds contain high seed oil, proteins, amino acids, and vitamin E, and are consumed worldwide as edible nut, peanut butter, or candy, and peanut oil extracted from the seeds. The meal remaining after oil extraction is also used for animal feed. However, its narrow germplasm base, together with susceptibility to diseases, pathogens, and weeds, decreases yield and seed quality and causes great economic losses annually. Hence, the optimization of efficient in vitro propagation procedures would be highly effective for peanut propagation, as it would raise yield and improve seed quality and flavor. Earlier reports on traditional micropropagation methods, based on axillary bud proliferation which guarantees the multiplication of true-to-type plants, are still limited. This chapter describes a micropropagation protocol to improve multiple shoot formation from shoot-tip explants by using AgNO(3) in combination with plant growth regulators.

Cryogenic technologies for the long-term storage of Citrus germplasm.

With its beautiful trees, Citrus species have long been valued by humanity. The tasteful fruits, extensively used for nutrition, are also good for health due to the high content in vitamins, minerals, and dietary fibers. Like majority of the woody fruit plants, Citrus germplasm is conserved mainly as field collections in clonal orchards. However, such a traditional approach presents several difficulties, among which are the high cost, manual labor, and extensive land required to maintain the collections, as well as the necessity of a careful protection of plants from diseases and extreme environmental conditions. As many species in the genus have seeds recalcitrant to desiccation, conservation in seed banks is also inadequate. On the other hand, cryopreservation, i.e., the storage of specimens at ultra-low temperatures (usually in liquid nitrogen, at -196°C) where reactions within the cells are minimized, presents a unique alternative for the safe storage of such germplasm. The present contribution outlines the cryopreservation techniques applied to seeds, zygotic and somatic embryos, embryogenic callus cultures of Citrus spp. and provides sample protocols to be used for Citrus conservation.

In vitro conservation and cryopreservation of ornamental plants.

Today, the conservation of ornamental germplasm can take advantage of innovative techniques which allow preservation in vitro (slow growth storage) or in liquid nitrogen (cryopreservation) of plant material. Slow growth storage refers to the techniques enabling the in vitro conservation of shoot cultures in aseptic conditions by reducing markedly the frequency of periodic subculturing, without affecting the viability and regrowth of shoot cultures. Cryopreservation refers to the storage of explants from tissue culture at ultra-low temperature (-196 degrees C). At such temperature, all the biological reactions within the cells are hampered, hence the technique makes available the storage of plant material for theoretically unlimited periods of time. An exhaustive review of papers dealing with the slow growth storage and the cryopreservation of ornamental species is reported here. Step-by-step protocols for the slow growth storage of rose germplasm, the production of synthetic seeds for the in vitro conservation of ornamentals, and the cryopreservation of Chrysanthemum morifolium are included.