In vitro study of the effect of NaCl on interspecific Populus L. hybrids

In conditions of climate aridisation and desertification of territories leading to secondary soil salinisation, more and more lands are subjected to premature degradation and their further withdrawal from agricultural use. In vitro modelling of stress factors for selection of tolerant lines of salt-tolerant genotypes of tree species helps to accelerate the breeding process. In the study, the effect of NaCl on morphogenesis of regenerant plants of poplar F1 interspecific hybrids (Populus L.) was investigated, and in vitro selection of potentially co-tolerant genotypes was carried out. After controlled crossing of selected parental lines with economically useful traits (Populus deltoides × P. alba; P. nigra f. piramidalis × P. alba; P. deltoides × P. bolleana f. piramidalis) by the method of isolated embryos, aseptic explants of interspecific hybrids were obtained and micropropagated. Sixteen genotypes of 10 clones each were tested on selective media with NaCl at concentrations of 0.2; 0.5;1.0 % in three replicates. The ratio of surviving and regenerating explants, the frequency of rhizogenesis and the percentage of chlorosis among the tested accessions, as well as shoot length and number, multiplication factor and number of nodes were determined.  The inhibitory effect of salt stress was manifested in the inhibition of morphogenesis, including the absence of rhizogenesis processes, chlorosis, partial deformation and necrosis of leaf plates. A dose of 0.5 % NaCl was determined to be sublethal and 1 % was lethal. Among the sixteen genotypes tested, 4 genotypes had high % of surviving explants (96.2–96.7 %), two of which maintained high % regeneration (82.6–96.6 %) at sublethal concentration of selective agent. These results suggest that in vitro testing for NaCl tolerance is a promising tool for accelerating the breeding process and selecting potentially tolerant genotypes to the abiotic stress factor for further field trials.

1. Mashkina O. S., Tabatskaya T. M., Korchagin O. M. In vitro selection of birch for tolerance to salinity stress. IOP Conference Series: Earth and Environmental Science. 2021;875(1):012082. DOI: https://doi.org/10.1088/1755-1315/875/1/012082

2. Kulczyk-Skrzeszewska M., Kieliszewska-Rokicka B. Influence of drought and salt stress on the growth of young Populus nigra ‘Italica’plants and associated mycorrhizal fungi and non-mycorrhizal fungal endophytes. New Forests. 2022;53(4):679–694. DOI: https://doi.org/10.1007/s11056-021-09879-6

3. Chen Y., Yuan B., Wei Z., Chen X., Chen Y., Qiu N. The ion homeostasis and ROS scavenging responses in ‘NL895’poplar plantlet organs under in vitro salinity stress. In Vitro Cellular & Developmental Biology-Plant. 2018;54:318–331. DOI: https://doi.org/10.1007/s11627-018-9896-z

4. Galović V., Kebert M., Popovic B., Kovacevic B., Vasić V., Joseph M., Orlović S., Szabados L. Biochemical and gene expression analyses in different poplar clones: The selection tools for afforestation of halomorphic environments. Forests. 2021;12(5):636. DOI: https://doi.org/10.3390/f12050636

5. Liu Y., Su M., Han Z. Effects of NaCl stress on the growth, physiological characteristics and anatomical structures of Populus talassica × Populus euphratica seedlings. Plants. 2022;11(22):3025. DOI: https://doi.org/10.3390/plants11223025

6. Singh D., Kumar A. In vitro screening and characterization of selected elite clones of Eucalyptus tereticornis Sm. for salt stress. Journal of Plant Growth Regulation. 2021;40:694–706. DOI: https://doi.org/10.1007/s00344-020-10138-9

7. Guleria I., Kumari A., Lacaille-Dubois M-A., Nishant, Kumar V., Saini A. K., Dhatwalia J., Lal S. A review on the genus Populus: A potential source of biologically active compounds. Phytochemistry Review. 2022;21:987–1046. DOI: https://doi.org/10.1007/s11101-021-09772-2

8. Erst A. A., Shishkin S. V., Voronkova M. S. The generation of interspecific hybrids in (Populus alba × P. bolleana) × P. × сanescens by in vitro culture. Sibirskiy lesnoy zhurnal = Siberian Journal of Forest Science. 2019;(2):45–52. (In Russ.). DOI: https://doi.org/10.15372/SJFS20190204

9. Erst A. A., Shishkin S. V., Voronkova M. S. Evaluation of salinity and osmotic stress resistance of the genus Populus species and hybrids in culture in vitro. Plant cell biotechnology and molecular biology. 2019;20(11-12):451–458. URL: https://www.researchgate.net/publication/334811183_EVALUATION_OF_SALINITY_AND_OS-MOTIC_STRESS_RESISTANCE_OF_THE_GENUS_Populus_SPECIES_AND_HYBRIDS_IN_CULTURE_In_vitro

10. Spivak V. V., Burmenko Yu. V., Kapitova I. A. Embryo rescue in stone fruit breeding: specifics and application success. Sadovodstvo i vinogradarstvo = Horticulture and viticulture. 2023;(5):27–34. (In Russ.). DOI: https://doi.org/10.31676/0235-2591-2023-5-27-34

11. Tabatskaya T. M., Mashkina O. S., Korchagin O. M. In vitro modelling of salinity stress for the selection of stress-tolerant birch lines. E3S Web of Conferences. 2020;224:04013. DOI: https://doi.org/10.1051/e3sconf/202022404013

12. Zholobova O. O., Mogilevskaya I. V., Melnik S. V. Screening Smoke Tree (Cotinus coggygria Scop.) on Osmotic Stress using Polyethylene Glycol 6000 in vitro. Indian Journal of Agricultural Research. 2024;58(1):36–42. DOI: https://doi.org/10.18805/IJARe.AF-781

13. Markova M. G., Somova E. N. Optimization of clonal micropropagation of stone fruit crops. Agrarnaya nauka Evro-Severo-Vostoka = Agricultural Science Euro-North-East. 2024;25(2):189–197. (In Russ.). DOI: https://doi.org/10.30766/2072-9081.2024.25.2.189-197

14. Vuksanovic V., Kovacevic B., Orlović S., Kebert M., Kovač M. The influence of drought on growth and development of white poplar shoots in vitro. Topola. Poplar. 2019;203:13–18. URL: https://ilfe.org/sites/default/files/04Vuksanovic_et_al.pdf

15. Vuksanović V., Kovacevic B., Kebert M., Katanic M., Kesic L., Karaklic V., Orlović S. In vitro modulation of antioxidant and physiological properties of white poplar induced by salinity. Glasnik Sumarskog fakulteta. 2019;120:179–196. DOI: https://doi.org/10.2298/GSF1920179V

16. Kryuchkov S. N., Solomentseva A. S., Romanenko A. K. Intraspecific crosses of selected biotypes of the Ulmus and Populus genera to obtain heterotic breeding forms. Agrarnyy vestnik Urala = Agrarian Bulletin of the Urals. 2023;23(6):23–34. (In Russ.). DOI: https://doi.org/10.32417/1997-4868-2023-235-06-23-34

17. Phillips G. C., Garda M. Plant tissue culture media and practices: an overview. In Vitro Cellular & Developmental Biology-Plant. 2019;55:242–257. DOI: https://doi.org/10.1007/s11627-019-09983-5

18. Jouve L., Hoffmann L., Hausman J. F. Polymine, carbohydrate, and proline content changesduring salt stress exposure of aspen (Populus tremula L.): involvement of oxidation and osmoregulation metabolism. Plant Biology. 2004;6(1):74–80. DOI: https://doi.org/10.1055/s-2003-44687

19. Fung L. E., Wang S. S., Altman A., Hütterman A. Effect of NaCl on growth, photosynthesis, ion and water relations of four poplar genotypes. Forest Ecology and management. 1998;107(1-3):135–146. DOI: https://doi.org/10.1016/S0378-1127(97)00328-9