Abstract
Introduction
In pine stands affected by annosum root rot (Heterobasidion annosum s.l.), there are Scots pine (Pinus sylvestris L.) trees that are able to maintain viability for a long time without showing external signs of the disease (“conditionally resistant”). Establishing forest stands with seeds from such trees can significantly increase the effectiveness of artificial reforestation. Scientists have noted that the growth and development of pine are to some extent determined by the cotyledons number in the germinants. Nowadays it is an urgent issue to develop methods for early diagnosis of hereditary properties in order to predict the characteristics of an adult plant.
The study aimed to analyse the possibility of using signs of cell mitotic activity of root apical meristems and a number of cotyledons in Scots pine germinating seedlings to predict the tree resistance to annosum root rot.
Materials and Methods
The study site is located in the northeastern part of Ukraine in the State Enterprise “Kharkiv Forest Research Station” (Kharkiv region). In the stand affected by root fungus, trees with different health conditions were selected, namely without external signs of decline (“conditionally resistant” – 13 trees), with signs of disease (“affected”– 7 trees) and control (5 trees). Cones were collected from the selected trees and seeds were removed. Seed samples of 100 seeds from each tree were taken. The sowing qualities of seeds (germination) and their weight were evaluated according to the methods generally accepted in forest seed production. We evaluated the mitotic activity of root cells and counted a number of cotyledons in germinating seedlings. Then, we analysed in variants the distribution of frequency of seedlings with different numbers of cotyledons.
Results
The seeds of “conditionally resistant” trees exceed those of “affected” and control trees by mass by 18.5% and 5%, respectively. There is no significant difference in the energy of seed germination between the seeds of trees with different resistance (65% and 64%, respectively). The proliferative activity of apical meristem cells of “conditionally resistant” trees is significantly higher than that of “affected” ones (tCt = 8.5; t0.01 = 4.0) and control (tCt = 5.1; t0.01 = 4.0). The biggest differences between germinating seedlings from the seeds of “conditionally resistant" and “affected” trees were found in the percentage of prophases (36.7% and 42.0%, respectively) and metaphases (32.7 and 25.7%, respectively). The number of cotyledons in germinants varied from three to nine. Germinating seedlings with six cotyledons were most often found in the samples. The total share of seeds whose seedlings had more than six cotyledons made 84.6% in “conditionally resistant” trees, 75.9% in “affected” trees and 80.71% in the control.
Conclusions
The seeds of “conditionally resistant” trees compared to “affected” ones are characterized by a larger average weight of 1,000 seeds and are not inferior in terms of their germination energy. The stability of the level of mitotic activity of cells in the roots of germinating seedlings of “conditionally resistant” trees as well as its greater intensity compared to “affected” ones were confirmed (MI = 10.7% and MI = 5.6%, respectively, where MI is mitotic index). Most often, the samples contain germinants with six cotyledons. The maximum total share of germinating seedlings in which a number of cotyledons were 6 or more was recorded for “conditionally resistant” trees. Based on the findings, we preliminarily recommend the use of the set above features for the diagnosis of resistance to annosum root rot among the young trees that have entered reproduction. Further research is necessary to definitely confirm or refute our hypothesis. The use of indicators of the mitotic activity of apical meristem cells and a number of cotyledons of germinating seedlings for the early assessment of Scots pine trees for their resistance to annosum root rot can increase the effectiveness of selection by these features. The application of cytological methods can be especially relevant in progeny tests, which should be established for the study of the inherited resistance traits.
3 Fig., 2 Tables, 27 Refs.
References
Аdomas, A., Heller, G., Li, G., Olson, ?., Chu, T. M., Osborne, J., Dean, R. A. 2007. Transcript profiling of a conifer pathosystem: response of Pinus sylvestris root tissues to pathogen (Heterobasidion annosum) invasion. Tree Physiology, 27 (10): 1441–1458.
Asiegbu, F. O., Choi, W.,Li, G., Nahalkova, J., Dean, R. A. 2003. Isolation of a novel antimicrobial peptide gene (Sp-AMP) homologue from Pinus sylvestris (Scots pine) following infection with the root rot fungus Heterobasidion annosum. FEMS microbiology letters, 228 (1): 27–31.
Castoldi, E., and Molina, J. A. 2014. Effect of seed mass and number of cotyledons on seed germination after heat treatment in Pinus sylvestris L. var. iberica Svob. Forest Systems, 23(3): 483–489. https://doi.org/10.5424/fs/2014233-05480
Chemeris, O. V. and Boyko, M. I. 2008. Content of phenolic compounds in seedlings of Pinus sylvestris L. and Pinus pallasiana D. Don infected by Heterobasidion annosum (Fr.) Bref. Problems of ecology and nature protection of technogenic region, 8: 267–272 (in Ukrainian).
Chernykh, A. G. 1965. Anatomical features of wood of pine individuals that survived in the foci of the annosum root rot]. Forestry and Forest Melioration, 7: 121–125 (in Russian).
Dyshko, V. A. and Torosova, L. O. 2016. Osoblyvosti rostovykh protsesiv sosny zvychaynoyi u nasadzheni, urazhenomu korenevoyu hubkoyu [Features of growth processes of Scots pine in plantation affected by annosum root rot]. Lisivnytstvo i ahrolisomelioratsiya [Forestry and Forest Melioration], 128: 134–142 (in Ukrainian).
Dyshko, V. A., Ustskyy, I. M., Mykhaylichenko O. A. 2015. Morphological and biochemical differences of trees with different resistance to annosum root rot. Forestry and Forest Melioration, 126: 218–224 (in Ukrainian).
Dyshko V. A., Ustsky I. M., Mykhaylichenko O. A. 2019. Features of morphological and biometric characteristics of reproductive organs of Scots pine trees with different resistance in stands affected by Heterobasidion annosum. Forestry and Forest Melioration, 135: 58–67(in Ukrainian). https://doi.org/10.33220/1026-3365.135.2019.58
Mar?iulynas, A., Sirgedait?-???ien?, V., ?emaitis, P., Baliuckas, V. 2019. The resistance of Scots pine (Pinus sylvestris L.) half-sib families to Heterobasidion annosum. Forests, 10(3): 287.
Nagy, N. E., Krokene, P., Solheim, H. 2006. Anatomical-based defense responses of Scots pine (Pinus sylvestris) stems to two fungal pathogens. Tree Physiology, 26(2): 159–167.
Negrutsky, S. F. 1986. Annosum root rot. Moscow, Agropromizdat, 196 p. (in Russian).
Orlenko, Ye. G. and Podzharova, Z. S. 1980. Early diagnostics of growth energy of Scots pine seedlings of different geographical origin. Lesovedeniye i Lesnoye khozyaystvo, 15: 39–43 (in Russian).
Poplavskaya, L. F. and Rebko, S. V. 2013. Breeding characteristics of Scots pine trees with different resistance to annosum root rot. In: Current state and prospects of conservation and protection of forests in the system of sustainable development. Proceedings of International Scientific and Practice Conf. Gomel, p. 310–314 (in Russian).
Popov, V. Ya., Tuchin, P. V., Fayzulin, D. Kh. 2002. Establishment of Scots pine plantations of seed origin on a breeding basis. Ecological problems of the North, 5: 72–85 (in Russian).
Popov, V. Ya. and Zharikov, V. M. 1973. The methods of selection and early diagnosis of hereditary properties of pine and spruce plus trees. Guidelines. Arkhangelsk, AIL i LKH, 40 p. (in Russian).
Popov, V. Ya. and Zharikov, V. M. 1977. The recommendations for the creation of permanent pine seed plantations on a tree improvement basis (for research and production testing). Arkhangelsk, AIL i LKH, 12 p. (in Russian).
Romanovskii, M. G. and Morozov, G. P. 2019. Cotyledons of seedlings and embryos of conifers. Russian Journal of Forest Science (Lesovedenie), 6: 573–579 (in Russian).
Sanitary Forests Regulations in Ukraine. 2016. [Electronic resource]. Resolution of the Cabinet of Ministers of Ukraine No 756 dated 26 October 2016. Available at: http://zakon2.rada.gov.ua/laws/show/555-95-п (accessed 15.09.2022) (in Ukrainian).
Seeds of trees and shrubs. Methods for seed testing (germination, viability, seed quality). DSTU 8558:2015 [State Standard of Ukraine]. 2017. Valid from 1 January 2017. Kyiv, DP UkrNDNTS, 91 p. (in Ukrainian).
Shoferistova, Ye. G. 1973. On the method of staining chromosomes and pollen. Botanical journal, 58 (7): 1011–1012 (in Russian).
Skipars, V. 2011. Genetic aspects of resistance of Scots pine (Pinus sylvestris L.) against root rot caused by Heterobasidion annosum (Fr.) Bref. Summary of the doctoral thesis for the scientific degree Dr. silv. in Forest Ecology and Silviculture.
Tereshchenko, L. I., Samoday, V. P., Los, S. A. 2011. The results of the study of the first in Ukraine progeny tests of Scots pine. Forestry and Forest Melioration, 118: 128–136 (in Ukrainian).
Torosova, L. O. 2008. Dynamics of mitotic activity of merystem cells of western larch (Larix occidentalis) needles. Forestry and Forest Melioration, 113: 206–209 (in Ukrainian).
Torosova, L. O. 2012. The mitotic activity of meristems of vegetative bud cells of the English oak (Quercus robur L.). Forestry and Forest Melioration, 120: 70–74 (in Ukrainian).
Ulusan, M. D. and Bilir, N. 2008. Broad-sense heritability for seedling characters and its importance for breeding in Scots pine. [Electronic resource]. Sd? fen edebiyat fak?ltesi fen dergisi (e-dergi), 3(2): 133–138. Available at: https://dergipark.org.tr/tr/download/article-file/116270 (accessed 15.09.2022).
Ustskiy, I. M. 2017. Features of root system structure in the foci of root rot caused by Heterobasidion annosum (Fr.) Bref. Forestry and Forest Melioration, 131: 187–193 (in Ukrainian).
Ustskiy, I. M., Dyshko, V. A., Mykhaylichenko, O. A. 2019. Peculiarities of seed germination and seedling growth of Scots pine trees with different root rot resistance. Forestry and Forest Melioration, 134: 154–161 (in Ukrainian). https://doi.org/10.33220/1026-3365.134.2019.154
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