Maritime pine ( Pinus pinaster Aiton) is a coniferous native of the Mediterranean basin. Because of its adaptability to a wide range of environmental conditions, the species have become a model for studies in coniferous forest management and functional genomics. Somatic embryogenesis (SE) has been so far, the preferred biotechnological strategy for maritime pine breeding programs initiated at the middle-end of the 20th century. To overcome the limitations of the induction and maturation phases in maritime pine SE, we analyzed the possible maternal influence on the embryogenic capability of megagametophytes from controlled crosses, as well as the effect of the temperature and water availability during SE process on the production of plants. A strong maternal effect on the embryogenic potential of maritime pine megagametophytes was observed in our experiments using half-sib and full-sib progenies, while paternal effect was almost undetectable. Besides, it seems possible to improve somatic embryo production of maritime pine megagametophytes by adjusting optimal temperature throughout the process: 28°C during induction and proliferation, and 23°C during the maturation phase. Using induction and proliferation media with reduced water availability (6 g/L Gelrite) can also increase embryo production. Since other limitation of maritime pine SE is culture decline of embryogenic masses (EMs), that reduces embryo yield and germination, we assessed the profile of ABA and IAA and the expression of two embryogenesis-related genes ( LEC1 and WOX2 ) during maturation of EMs of two morphotypes that differed in their maturation capability. Spiky morphotype (SK), with high maturation capability, had a steady increase in both hormones along the 12 weeks of the maturation, whereas ABA content in smooth morphotype picked at the 4th week and dropped. EMs with this morphotype also had a higher IAA content at the beginning of the maturation. A decrease of LEC1 and WOX2 gene expression over the course of embryo development was found to be characteristic of the SK with high maturation capability.

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Rooting space is considered as a resource in plants, but comparative studies on the biomass allocation plasticity in response to rooting volume (RV) are rare. We compared responses in growth, biomass allocation and ontogenetic heteroblasty in nine hard pine species of contrasted ecology. Seedlings were cultivated in containers of 0.2, 0.5, 1.0, 2.8 and 7 L for two growing seasons (425 days). Reduction in RV caused a reduction in plant absolute and relative growth rate and biomass allocated to stems but it increased biomass allocated to roots. RV affected to a lesser extent and in a less consistent direction allocation to leaves. Species that grew faster (higher relative growth rate) had a steeper decrease in growth with the reduction in RV. Ontogenetic heteroblasty, evaluated as the proportion of secondary needles in the needle biomass, showed highly different plasticity patterns in response to RV. Decrease in RV caused negligible or no change either in the most ontogenetically delayed Mediterranean pines or in the most ontogenetically advanced pines, the mesic Pinus sylvestris and P. uncinata. By contrast, ontogenetically intermediate species showed steep reaction norms in response to reduction in RV. While P. pinaster and P. brutia showed marked rejuvenation, P. nigra accelerated the development of adult foliage.

Compared to natural selection, domestication implies a dramatic change in traits linked to fitness. A number of traits conferring fitness in the wild might be detrimental under domestication, and domesticated species typically differ from their ancestors in a set of traits known as the domestication syndrome. Specifically, trade-offs between growth and reproduction are well established across the tree of life. According to allocation theory, selection for growth rate is expected to indirectly alter life-history reproductive traits, diverting resources from reproduction to growth. Here we tested this hypothesis by examining the genetic change and correlated responses of reproductive traits as a result of selection for timber yield in the tree Pinus pinaster. Phenotypic selection was carried out in a natural population, and progenies from selected trees were compared with those of control trees in a common garden experiment. According to expectations, we detected a genetic change in important life-history traits due to selection. Specifically, threshold sizes for reproduction were much higher and reproductive investment relative to size significantly lower in the selected progenies just after a single artificial selection event. Our study helps to define the domestication syndrome in exploited forest trees and shows that changes affecting developmental pathways are relevant in domestication processes of long-lived plants.

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