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. 2022 Apr 1;11(7):965.
doi: 10.3390/plants11070965.

Effect of Sucrose on Growth and Stress Status of Castanea sativa x C. crenata Shoots Cultured in Liquid Medium

Diego Gago  1   2 , María Ángeles Bernal  2 , Conchi Sánchez  1 , Anxela Aldrey  1 , Beatriz Cuenca  3 , Colin Bruce Christie  4 , Nieves Vidal  1
Affiliations

Effect of Sucrose on Growth and Stress Status of Castanea sativa x C. crenata Shoots Cultured in Liquid Medium

Diego Gago et al. Plants (Basel). .

Abstract

Current breeding programs aim to increase the number of ink-tolerant chestnut trees using vegetative propagation of selected genotypes. However, the commercial vegetative propagation of chestnut species is still a bottleneck for the forest industry, mainly due to problems in the rooting and acclimation of propagules. This study aimed to explore the potential benefits of decreasing sucrose supplementation during chestnut micropropagation. Explants were cultured with high light intensity and CO2-enriched air in temporary or continuous immersion bioreactors and with different sucrose supplementation to evaluate the impact of these treatments on growth, rooting and physiological status (monosaccharide content, soluble phenolics and antioxidant activity). The proliferation and rooting performance of shoots cultured by continuous immersion decreased sharply with sucrose concentrations lower than 1%, whereas shoots cultured by temporary immersion grew and rooted successfully with 0.5% sucrose. These results suggest this system is appropriate to culture chestnut with low sucrose concentration and to explore photoautotrophic propagation of this species.

Keywords: bioreactors; chestnut; continuous immersion; photoautotrophy; photosynthesis; temporary immersion.

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Conflict of interest statement

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

Figures

Figure 1
Figure 1
Chestnut shoots of clone CO53 after 8 weeks of culture by continuous immersion with 0.5% sucrose (a), 1% sucrose (b) and 3% sucrose (c).
Figure 2
Figure 2
Chestnut shoots of clone PO42 after 8 weeks of culture by temporary immersion with 0.5% sucrose (a), 1% sucrose (b) and 3% sucrose (c).
Figure 3
Figure 3
Effect of sucrose supplementation (0.5, 1 and 3%) on proliferation rates of apical sections of clone CO53 shoots cultured by continuous immersion. (a) Number of shoots (NS). (b) Length of the longest shoot (SL) and length (LL) and width (LW) of the largest leaf per explant. Values are the mean ± standard error from two replicate trials each with 2 bioreactors (64 explants/treatment). For each variable, different letters indicate significant differences at p < 0.05.
Figure 4
Figure 4
Effect of sucrose supplementation (0.5, 1 and 3%) on proliferation rates of apical sections of clone PO42 shoots cultured by temporary immersion. (a) Number of shoots (NS). (b) Length of the longest shoot (SL) and length (LL) and width (LW) of the largest leaf per explant. Values are the mean ± standard error from three replicate trials each with 2 bioreactors (72 explants/treatment). For each variable, different letters indicate significant differences at p < 0.05.
Figure 5
Figure 5
Chestnut shoots from clones CO53 and PO42 multiplied and rooted with different sucrose concentrations. (a) Shoots of CO53 cultured in sucrose 3% and rooted in sucrose 0.5, 1 and 3%. (b) Shoots of CO53 cultured in sucrose 0.5% and rooted in sucrose 0.5, 1 and 3%. (c) Shoots of PO42 cultured in sucrose 3% and rooted in sucrose 0.5, 1 and 3%. (d) Shoots of PO42 cultured in sucrose 0.5% and rooted in sucrose 0.5, 1 and 3%. Photos taken 5 weeks after root induction. S0.5, S1 and S3: Sucrose 0.5, 1 and 3%.
Figure 6
Figure 6
Effect of sucrose supplementation on monosaccharide content of shoots of clones PO42 and CO53 cultured by temporary and continuous immersion, respectively. Values are the mean ± standard error from 24 shoots analyzed independently. Different capital letters indicate significant differences in relation to the genotype, and different lowercase letters indicate significant differences in relation to the sucrose supplementation (p < 0.05). GE, Glucose equivalents.
Figure 7
Figure 7
Effect of sucrose supplementation on antioxidant activity of shoots of clones PO42 and CO53 cultured by temporary and continuous immersion, respectively. Values are the mean ± standard error from 24 shoots analyzed independently. Different capital letters indicate significant differences in relation to the genotype, and different lowercase letters indicate significant differences in relation to the sucrose supplementation (p < 0.05). TE: Trolox equivalents.
Figure 8
Figure 8
Effect of sucrose supplementation on soluble phenolic compounds of shoots of clones PO42 and CO53 cultured by temporary and continuous immersion, respectively. Values are the mean ± standard error from 24 shoots analyzed independently. Different capital letters indicate significant differences in relation to the genotype, and different lowercase letters indicate significant differences in relation to the sucrose supplementation (p < 0.05). GAE: Gallic acid equivalents.
Figure 9
Figure 9
Rooting of chestnut shoots from clones CO53 and PO42. (a) Shoots of CO53 cultured in S3% selected for rooting. (b,c) Shoots of CO53 cultured in S1% during root induction (b) and inserted in rockwool cubes. (c,d) Shoots of PO42 cultured in S1% during root induction. (e,f) Shoots of CO53 cultured in S1% and transferred to medium with S0.5, 1 and 3% 24 h after root induction.
Figure 9
Figure 9
Rooting of chestnut shoots from clones CO53 and PO42. (a) Shoots of CO53 cultured in S3% selected for rooting. (b,c) Shoots of CO53 cultured in S1% during root induction (b) and inserted in rockwool cubes. (c,d) Shoots of PO42 cultured in S1% during root induction. (e,f) Shoots of CO53 cultured in S1% and transferred to medium with S0.5, 1 and 3% 24 h after root induction.
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