High gellan gum concentration and secondary somatic embryogenesis: two key factors to improve somatic embryo development in Pseudotsuga menziesii [Mirb.]

Lelu‐Walter, Marie‐Anne; Gautier, Florian; Eliášová, Kateřina; Sanchez, Léopoldo; Teyssier, Caroline; Lomenech, Anne-Marie; Metté, Claire Le; Hargreaves, Cathy; Trontin, Jean-François; Reeves, Cathie

doi:10.1007/s11240-017-1318-0

Cited by 34 publications

(40 citation statements)

References 53 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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“…Studies showed that high gellan gum concentrations improved in vitro germination frequencies in Pinus strobus L. embryos [25] and somatic embryo development in Pseudotsuga menziesii (Mirb.) Franco [26]. In vivo application of native gellan gum and its derivative enhanced biomass growth, photosynthesis intensity, and mineral content in Eucomis bicolor L'Hér.…”

Section: Introductionmentioning

confidence: 99%

Effects of Depolymerized Gellan with Different Molecular Weights on the Growth of Four Bedding Plant Species

Salachna

2020

Agronomy

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New solutions allowing for the shortening of the growing cycle and improvements in plant quality are constantly sought in order to improve the efficiency of bedding plant production under covers. Biodegradable polysaccharides and their derivatives have become increasingly popular in horticulture as plant growth promoters. A greenhouse pot experiment was conducted to evaluate the effects of depolymerized gellan of different molecular weights (MW 56 kDa and 77 kDa) on the growth and physiological parameters of ornamental bedding plants Rudbeckia hirta L., Salvia splendens Sellow ex J.A. Schultes, Scabiosa atropurpurea L., and Tithonia rotundifolia (Mill.) S.F. Blake. The results showed that the application of depolymerized gellan accelerated flowering and stimulated the growth of all assessed species, regardless of MW. The plants treated with depolymerized gellan grew higher and had greater fresh weight of their above-ground parts, higher leaf relative chlorophyll content (SPAD; soil and plant analysis development), and higher stomatal conductance (gs). The use of 56 kDa gellan fraction resulted in the formation of inflorescences with the greatest fresh weight in S. atropurpurea. Leaves of R. hirta treated with this fraction showed the highest values of SPAD and gs. This study demonstrated that gellan derivatives of low MW may be used for the production of innovative plant biostimulants.

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“…Somatic embryogenesis techniques for propagating Douglas-fir ( Pseudotsuga menziesii (Mirb) Franco), a productive conifer species of the Pinaceae family with globally appreciated wood quality, have been under development for more than 30 years [ 4 ]. There is a wealth of patented methods, but some recent improvements for steps from initiation to efficient production of somatic seedlings have just been made publicly available ([ 5 , 6 ], and references therein).…”

Section: Introductionmentioning

confidence: 99%

“…SE clusters of some conifer species, such as Douglas-fir, develop into polyembryogenic center of various sizes, putatively through continuous but incomplete cleavage polyembryony (somatic polyembryogenesis) and /or de novo somatic embryogenesis from proliferating early SEs [ 6 ]. In addition, some Douglas-fir lines of proliferating EMs contain both immature SEs and clusters of non-embryogenic cells (NECs) [ 5 , 16 ].…”

Section: Introductionmentioning

confidence: 99%

“…Subsequent transition from early SEs to cotyledonary SEs is stimulated when EMs are exposed to specific maturation conditions. Usual requirements for this transition, inter alia for Douglas-fir cultures [ 6 ], are supplementation of the medium with abscisic acid (ABA) in conjunction with increases in osmotic pressure (using a solution with high carbohydrate concentration, such as 0.2 M sucrose), and/or reduction of the water potential using high molecular weight polymers (such as 4000 Da polyethylene glycol, PEG 4000) or physically reducing water availability for the cultured cells by increased the medium’s gel strength.…”

Section: Introductionmentioning

confidence: 99%

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Repetitive somatic embryogenesis induced cytological and proteomic changes in embryogenic lines of Pseudotsuga menziesii [Mirb.]

et al. 2018

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BackgroundTo explore poorly understood differences between primary and subsequent somatic embryogenic lines of plants, we induced secondary (2ry) and tertiary (3ry) lines from cotyledonary somatic embryos (SEs) of two Douglas-fir genotypes: SD4 and TD17. The 2ry lines exhibited significantly higher embryogenic potential (SE yields) than the 1ry lines initiated from zygotic embryos (SD4, 2155 vs 477; TD17, 240 vs 29 g− 1 f.w.). Moreover, we observed similar differences in yield between 2ry and 3ry lines of SD4 (2400 vs 3921 g− 1 f.w.). To elucidate reasons for differences in embryogenic potential induced by repetitive somatic embryogenesis we then compared 2ry vs 1ry and 2ry vs 3ry lines at histo-cytological (using LC-MS/MS) and proteomic levels.ResultsRepetitive somatic embryogenesis dramatically improved the proliferating lines’ cellular organization (genotype SD4’s most strongly). Frequencies of singulated, bipolar SEs and compact polyembryogenic centers with elongated suspensors and apparently cleavable embryonal heads increased in 2ry and (even more) 3ry lines. Among 2300–2500 identified proteins, 162 and 228 were classified significantly differentially expressed between 2ry vs 1ry and 3ry vs 2ry lines, respectively, with special emphasis on “Proteolysis” and “Catabolic process” Gene Ontology categories. Strikingly, most of the significant proteins (> 70%) were down-regulated in 2ry relative to 1ry lines, but up-regulated in 3ry relative to 2ry lines, revealing a down-up pattern of expression. GO category enrichment analyses highlighted the opposite adjustments of global protein patterns, particularly for processes involved in chitin catabolism, lignin and L-phenylalanine metabolism, phenylpropanoid biosynthesis, oxidation-reduction, and response to karrikin. Sub-Network Enrichment Analyses highlighted interactions between significant proteins and both plant growth regulators and secondary metabolites after first (especially jasmonic acid, flavonoids) and second (especially salicylic acid, abscisic acid, lignin) embryogenesis cycles. Protein networks established after each induction affected the same “Plant development” and “Defense response” biological processes, but most strongly after the third cycle, which could explain the top embryogenic performance of 3ry lines.ConclusionsThis first report of cellular and molecular changes after repetitive somatic embryogenesis in conifers shows that each cycle enhanced the structure and singularization of EMs through modulation of growth regulator pathways, thereby improving the lines’ embryogenic status.Electronic supplementary materialThe online version of this article (10.1186/s12870-018-1337-y) contains supplementary material, which is available to authorized users.

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High gellan gum concentration and secondary somatic embryogenesis: two key factors to improve somatic embryo development in Pseudotsuga menziesii [Mirb.]

Cited by 34 publications

References 53 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

Effects of Depolymerized Gellan with Different Molecular Weights on the Growth of Four Bedding Plant Species

Repetitive somatic embryogenesis induced cytological and proteomic changes in embryogenic lines of Pseudotsuga menziesii [Mirb.]

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