Effect of partial drying and desiccation on somatic seedling quality in Norway and Serbian spruce

Hazubska-Przybył, Teresa; Wawrzyniak, Mikołaj Krzysztof; Obarska, A.; Bojarczuk, K.

doi:10.1007/s11738-014-1735-1

Cited by 13 publications

(11 citation statements)

References 32 publications

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“…Many studies have investigated how the culture conditions during the maturation phase influence a somatic embryo's capability to germinate, including factors such as, osmoticum (Bozhkov and von Arnold 1998; Hazubska-Przybył and Wawrzyniak 2017), abscisic acid (Bozhkov and von Arnold 1998;Montalbán et al 2010;Alvarez et al 2013), redox agents (Stasolla et al 2004;Pullman et al 2015), gelling agents (Klimaszewska et al 2000;Morel et al 2014a;Lelu-Walter et al 2018) and desiccation treatments (Hazubska-Przybył et al 2015;Carneros et al 2017). In terms of N metabolism, studies have suggested that a reduced accumulation of storage protein during maturation can have a negative impact on the quality of the mature somatic embryos, as compared with zygotic embryos (Klimaszewska et al 2004(Klimaszewska et al , 2016Morel et al 2014b).…”

Section: Electronic Supplementary Materialsmentioning

confidence: 99%

Nitrogen utilization during germination of somatic embryos of Norway spruce: revealing the importance of supplied glutamine for nitrogen metabolism

Carlsson

Egertsdotter

Ganeteg

et al. 2018

Trees

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Key message This paper shows that germinating Norway spruce somatic embryos are dependent on the carbon and nitrogen supplied in the medium, and that supplied glutamine accounts for 50 % of assimilated nitrogen during germination. Abstract The female megagametophyte, which provides the zygotic embryo with nitrogen (N), carbon (C) and energy during germination, is not present in Norway spruce (Picea abies) mature somatic embryos. Therefore, somatic embryos presumably rely on nutrients supplied in the germination medium in addition to their storage compounds accumulated during maturation. However, to what extent stored versus supplied compounds contribute to a somatic embryo germination is unclear. In this 24-day study, we addressed the above question by monitoring the biomass changes and the N and C budget during somatic embryo germination, under low-intensity red light. We found that the C and N storage reserves, accumulated during the maturation phase, were not sufficient to support the growth of the germinating somatic embryos, rather they were dependent on the medium components. In addition, in a previous study it has been found that glutamine (Gln) supplied in the medium was crucial for maintaining the primary amino acid (AA) metabolism and growth of the proliferating embryogenic cultures of Norway spruce (Carlsson et al., PLoS One 12(8):e0181785, 2017). Therefore, we hypothesised that Gln would be required as a significant source of N also during somatic embryo germination. By tracing the uptake of isotopically labelled N-sources from the medium and further into primary N assimilation, we found that Gln was the preferred source of N for the germinating somatic embryos, accounting for 50% of assimilated N. As the amounts of both arginine (Arg) and Gln were increased in the germinating somatic embryos, it also suggested that germination in low-intensity red light promoted N storage, similar to what has been observed in the zygotic embryo maturation in conifers (King, Gifford, Plant Physiol 113:1125-1135, 1997).

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Section: Electronic Supplementary Materialsmentioning

confidence: 99%

Nitrogen utilization during germination of somatic embryos of Norway spruce: revealing the importance of supplied glutamine for nitrogen metabolism

Carlsson

Egertsdotter

Ganeteg

et al. 2018

Trees

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“…In the case of Norway spruce, high initiation rates and recovery of embryogenic tissues from cryopreservation have been observed in previous studies (see, e.g., Nørgaard et al 1993;Klimaszewska et al 2001b;Varis et al 2017). The efficiency of somatic embryogenesis has been greatly improved, e.g., by initiating embryogenic tissues from immature zygotic embryos, manipulating the concentration of plant growth regulators and chemical composition of the culture medium (Hakman et al 1985;Klimaszewska and Smith 1997;Bozhkov and von Arnold 1998;Stasolla and Yeung 2003;Klimaszewska et al 2007;Hazubska-Przybył et al 2015;Varis et al 2017). Various methods concerning the germination, acclimation and selection of emblings have been introduced to improve propagation efficiency (Högberg et al 2001(Högberg et al , 2003Varis et al 2014;Llebrés et al 2018a, b).…”

Section: Introductionmentioning

confidence: 76%

“…In addition, the higher survival of shorter cotyledonary embryos in Paper I may also be an indication of too long a maturation period, which has been reported to decrease embling survival in later phases of SE (Morel et al 2014). Shorter maturation treatments have already been applied in different SE protocols in Norway spruce (see, e.g., Bozhkov and von Arnold 1998;Högberg et al 2001;Hazubska-Przybył et al 2015).…”

Section: Yield Survival and Height Growth After Improvements (Papersmentioning

confidence: 99%

Improved propagation efficiency in a laboratory–nursery interface for somatic embryogenesis in Norway spruce

Tikkinen¹

2018

Diss. For.

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The aim of this work was to improve the protocol of somatic embryogenesis (SE) and propagation efficiency in Norway spruce (Picea abies (L.) Karst.), which would enable the integration of SE into Finnish breeding programme and the nursery practices applied to seedlings. The studies specifically investigated the following three areas: i) how maturation, cold storage, germination and growing conditions (laboratory-nursery interface) affect the survival and height growth of emblings (Papers I and II); ii) how to improve the efficiency of embling production from genotypes from wide genetic backgrounds (Papers I and II); and iii) how to increase propagation efficiency by rooting cuttings from emblings, and produce field testing material by combining SE and the rooting of cuttings (Papers II and III). To evaluate the possibility of improving the efficiency of SE in the laboratory-nursery interface, a series of experiments were conducted. The cost structure of SE, and the effects of improvements on costs, was estimated. As a result, the protocol improvements doubled the yield of cotyledonary embryos, nearly doubled embling survival, and increased the height growth of emblings in the nursery by so much that sufficient planting height was reached one year less than before. Emblings were also obtained from 356 genotypes (50% thawed), and embling cuttings rooted well in conditions similar to those used for seedling cuttings. The protocol improvements also reduced embling production costs by 75%. Based on this work, emblings may be grown in nurseries after one week of in vitro germination, without any measures that differ from seedlings after transplanting. Propagation efficiency may be further increased by rooting embling cuttings. Furthermore, large-scale clone testing can be initiated with 5-12 emblings acting as cutting donors.

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“…These changes are associated with the changes in water content, which could be of interest for further studies because the fact that partial drying of somatic embryos at high relative humidity or desiccation in the presence of supersaturated solutions of salt is used to improve the efficiency of germination. Hazubska-Przybyl et al found that ESEs of Norway Spruce used in this study react on the heat stress and their behaviour differed from other ESEs [ 29 ]. During the first four weeks of cultivation, the ESE cluster seemed to be compactly aggregated; thereafter, the differences between the 1 H nuclei concentration in the inner and outer clusters were more evident.…”

Section: Resultsmentioning

confidence: 99%

Are Early Somatic Embryos of the Norway Spruce (Picea abies (L.) Karst.) Organised?

et al. 2015

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BackgroundSomatic embryogenesis in conifer species has great potential for the forestry industry. Hence, a number of methods have been developed for their efficient and rapid propagation through somatic embryogenesis. Although information is available regarding the previous process-mediated generation of embryogenic cells to form somatic embryos, there is a dearth of information in the literature on the detailed structure of these clusters.Methodology/Principal FindingsThe main aim of this study was to provide a more detailed structure of the embryogenic tissue clusters obtained through the in vitro propagation of the Norway spruce (Picea abies (L.) Karst.). We primarily focused on the growth of early somatic embryos (ESEs). The data on ESE growth suggested that there may be clear distinctions between their inner and outer regions. Therefore, we selected ESEs collected on the 56th day after sub-cultivation to dissect the homogeneity of the ESE clusters. Two colourimetric assays (acetocarmine and fluorescein diacetate/propidium iodide staining) and one metabolic assay based on the use of 2,3,5-triphenyltetrazolium chloride uncovered large differences in the metabolic activity inside the cluster. Next, we performed nuclear magnetic resonance measurements. The ESE cluster seemed to be compactly aggregated during the first four weeks of cultivation; thereafter, the difference between the 1H nuclei concentration in the inner and outer clusters was more evident. There were clear differences in the visual appearance of embryos from the outer and inner regions. Finally, a cluster was divided into six parts (three each from the inner and the outer regions of the embryo) to determine their growth and viability. The innermost embryos (centripetally towards the cluster centre) could grow after sub-cultivation but exhibited the slowest rate and required the longest time to reach the common growth rate. To confirm our hypothesis on the organisation of the ESE cluster, we investigated the effect of cluster orientation on the cultivation medium and the influence of the change of the cluster’s three-dimensional orientation on its development. Maintaining the same position when transferring ESEs into new cultivation medium seemed to be necessary because changes in the orientation significantly affected ESE growth.Conclusions and SignificanceThis work illustrated the possible inner organisation of ESEs. The outer layer of ESEs is formed by individual somatic embryos with high metabolic activity (and with high demands for nutrients, oxygen and water), while an embryonal group is directed outside of the ESE cluster. Somatic embryos with depressed metabolic activity were localised in the inner regions, where these embryonic tissues probably have a very important transport function.

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Effect of partial drying and desiccation on somatic seedling quality in Norway and Serbian spruce

Cited by 13 publications

References 32 publications

Nitrogen utilization during germination of somatic embryos of Norway spruce: revealing the importance of supplied glutamine for nitrogen metabolism

Nitrogen utilization during germination of somatic embryos of Norway spruce: revealing the importance of supplied glutamine for nitrogen metabolism

Improved propagation efficiency in a laboratory–nursery interface for somatic embryogenesis in Norway spruce

Are Early Somatic Embryos of the Norway Spruce (Picea abies (L.) Karst.) Organised?

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