1. Propagation, conservation, and improvement of forest species

Its objective is the development of protocols to propagate endangered forest species (in agreement with the Generalitat Valenciana) or those genotypes/families that show exceptional growth and/or production characteristics. These have been applied to Fraxinus ornus, Sorbus domestica, Robinia pseudoacacia, Cedrus atlantica, and Cedrus libani.

Since 2005, the group has been working on optimizing somatic embryogenesis protocols in Pinus pinaster (maritime pine) and Quercus ilex (holm oak). In Pinus pinaster, somatic embryogenesis has been induced from explants from three origins (Serra Calderona, Soria-Burgos, and Galicia-Costa), and plants have been obtained from some of the regenerated lines. The effect of liquid medium and temperature during the different phases of the process is being studied. In Quercus ilex, the effect of explant age, male and female flowers, on embryogenic capacity has been determined (MICINN). As a result of collaboration with the University of Málaga, University of Alcalá de Henares, and INIA, the group is participating in a complementary action to obtain haploid lines of maritime pine.

2.  Application of in vitro culture in forest genomics

• Validation of genes responsible for growth characteristics and drought stress tolerance.

Started in 2010 (European Plant-KBBE project), the objective is to use transgenic maritime pine plants to validate candidate genes involved in growth responses. From embryogenic lines, transgenic plants are being generated with varying levels of expression (overexpression or silencing via RNAi) of the genes responsible for these traits. This project involves laboratories from France, Spain, Germany, and Portugal.

• Obtaining haploid lines.

Currently (PROCOGEN Project, European part funded by the EU), haploid lines of maritime pine are being generated to be used as starting material for sequencing the genome of this species. This is a project involving European, North American (USA and Canada), and private company groups.

3. Genetic improvement of species of horticultural and medicinal interest

Our group developed a protocol for the propagation of Nerium oleander (oleander) varieties with ornamental interest. Based on this knowledge, in 2009 we signed a contract with the company SAT Even 46 CV to sanitize oleander varieties and produce bacteria-free mother plants to be used in their campaigns. This contract is renewed annually. Protocols have been developed for the genetic transformation of species with pharmaceutical interest (Digitalis minor) and cosmetic interest (Lavandula latifolia, spike lavender) (1996-2008, University of Valencia), which have been used to introduce genes of interest into Digitalis and serve as the basis for line 4.

4. Metabolic engineering of terpenes

The objective is to study the mechanisms that regulate the synthesis and accumulation of terpenes in the glandular trichomes of spike lavender (PROMETEU program). We have overexpressed the Arabidopsis genes HMG1 (the enzyme catalyzes the first reaction of the cytoplasmic MVA pathway), DXS (the enzyme catalyzes the first reaction of the MEP pathway, chloroplastic), and LS from Mentha spicata (encodes Limonene synthase, LS, which catalyzes the conversion of geranyl diphosphate to limonene), reaching the following conclusions:

1. Overexpression of both DXS and HMG1 significantly increases essential oil production.
2. Apparently, the MVA pathway contributes to the biosynthesis of the terpenoid constituents of lavender oil. We are currently studying whether this contribution results from cytoplasmic IPP being normally transferred to plastids in non-transformed plants.
3. Overexpression of the LS gene modifies the monoterpene profile of lavender oil, mainly in young leaves.