Geographically distant Scots pine (Pinus sylvestris L.) populations are adapted to specific photoperiods and temperature gradients, and markedly vary in the timing of growth patterns and adaptive traits. To understand the variability of adaptive capacity within species, molecular mechanisms that govern the physiological aspects of phenotypic plasticity should be addressed. Protein expression analysis is capable of depicting molecular events closely linked to phenotype formation. Therefore, in this study, we used comparative proteomics analysis to differentiate Scots pine genotypes originating from geographically distant populations in Europe, which show distinct growth and cold adaptation phenotypes. Needles were collected from 3-month-old seedlings originating from populations in Spain, Lithuania and Finland. Under active growth-promoting conditions and upon acclimation treatment, 65 and 53 differentially expressed proteins were identified, respectively. Constitutive protein expression differences detected during active growth were associated with cell metabolism and stress response, and conveyed a population-specific adaptation to the distinct climatic conditions. Acclimation-induced protein expression patterns suggested the presence of a similar cold adaptation mechanism among the populations. Variation of adaptive capacity among the genotypes was potentially represented by a constitutive low level of expression of the Ser/Thr-protein phosphatase, the negative regulator of the adaptive response. Also, overall less pronounced acclimation-induced response in seedlings from the Spanish population was observed. Thus, our study demonstrates that comparative proteomic analysis of young conifer seedlings is capable of providing insights into adaptation processes at the cellular level, which could help to infer variability of adaptive capacity within the plant species.
Scots pine is economically and ecologically important forest tree species in Lithuania. According to the Regulation of Forest Planting and Restoration of Lithuania ( 2018) and the Regulations on Forest Reproductive Material (FRM) (2017) forests must be planted and replanted with the target tree species, using high quality seeds and seedlings from a given provenance. EU Council Directive (1999/105/EC) indicates that EU Member States are responsible for the regulation, requirements and implementation of FRM quality and traceability system. The existing FRM control system in Lithuania is based on the control actions of the supporting documents regarding the origin of FRM. However, this FRM traceability system is limited and, in some cases, cannot ensure that FRM used for reforestation/afforestation originates from the declared place of origin or belongs to a certain seed or plant lot. In this study we aimed to assess and compare genetic diversity of scots pine seed orchard clones with seeds, and to test DNA based parentage analysis methods with Jonava scots pine seed orchard clones and seeds. Up to now DNA markers based FRM traceability system in Lithuania has not been tested and implemented. Thus, our first results based on 12 nuclear microsatellite markers indicated that genetic diversity among Jonava seed orchard clones and collected seeds were moderately high and comparable with other studies on scots pine in Europe. The test of the maternity analysis software CERVUS has showed very positive results in maternity assignments. Therefore, selected microsatellite markers in combination with accurate sampling design and specific DNA analysis methods should be improved and can be used for FRM traceability system based on DNA "fingerprints". This work is supported by the Project. No. 09.3.3-LMT-K-712-19-0007.
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