Primula veris plants derived from in vitro cultures and from seeds: genetic stability, morphology, and seed characteristics
Flow cytometry measurement confirmed the genetic stability of Primula veris L. (cowslip) plants obtained during the micropropagation process. The mean 2C DNA content was 0.97 pg/2C and P. veris can be classified in the group of plants of very small genomes. Comparative morphological analysis of cowslip plants regenerated in vitro and derived from seeds was carried out in three subsequent years under common garden conditions, which limited the effects of environmental factors. The origin of specimens significantly influenced most of the morphological traits examined. Plants that originated from seeds were characterized by more intensive development rate and higher total seed production compared to micropropagated plants. Significant differences between the two groups of specimens were observed for the number of flower stalks, flowers, fruits, and seeds per plant. No interseasonal variation within the reproductive traits such as number of seeds per flower stalk and per plant was found for plants derived from in vitro cultures. That allows prediction of seed production from in vitro regenerated plants after their successful acclimatization to field conditions. Importantly, plants derived from in vitro cultures set significantly bigger seeds. Our results are promising for broad uses of the regenerated in vitro cowslip plants in ornamental horticulture and the pharmaceutical industry.
___
- Ambika S, Manonmani V, Somasundaram G (2014). Review on
effect of seed size on seedling vigour and seed yield. Research
Journal of Seed Science 7: 31-38.
- Antrobus S, Lack AJ (1993). Genetics of colonizing and established
populations of
Primula veris
. Heredity 71: 252-258.
- Bajaj YPS, Furmanova M, Olszowska O (1988). Biotechnology of
the micropropagation of medicinal and aromatic plants. In:
Bajaj YPS, editor. Biotechnology in Agriculture and Forestry,
Medicinal and Aromatic Plants, Vol. 4. 1st ed. Berlin, Germany:
Springer-Verlag, pp. 60-103.
- Baker K, Richards AJ, Tremayne M (1994). Fitness constraints on
flower number, seed number and seed size in the dimorphic
species
Primula farinosa
L. and
Armeria maritima
(Miller)
Wild. New Phytol 128: 563-570.
- Baskin CC, Baskin JM (2014). Seeds: Ecology, Biogeography, and
Evolution of Dormancy and Germination. 2nd ed. San Diego,
CA, USA: Academic Press.
- Berisha N, Millaku F, Gashi B, Krasniqi E, Novak J (2015). Initial
determination of DNA polymorphism of some
Primula veris
L. populations from Kosovo and Austria. Physiol Mol Biol Pla
21: 117-122.
- Bhatia P, Ashwath N (2004). Comparative performance of
micropropagated and seed-grown tomato plants. Biol
Plantarum 48: 625-628.
- Boyd M, Silvertown J, Tucker C (1990). Population ecology of
heterostyled and homostyle
Primula vulgaris
: growth, survival
and reproduction in field populations. J Ecol 78: 799-813.
- Brys R, Jacquemyn H (2009). Biological flora of the British Isles:
Primula veris
L. J Ecol 97: 581-600.
- Budzianowski J, Morozowska M, Wesołowska M (2005). Lipophilic
flavones of
Primula veris
L. from field cultivation and
in vitro
cultures. Phytochemistry 66: 1033-1039.
- Cassells AG, Curry RF (2001). Oxidative stress and physiological
epigenetic and genetic variability in plant tissue culture:
implications for micropropagators and genetic engineers. Plant
Cell Tiss Org 64: 145-157.
- Clapham AR, Tutin TG, Moore DM (1987). Flora of the British Isles.
3rd ed. Cambridge, UK: Cambridge University Press.
- Doležel J, Bartos J (2005). Plant DNA flow cytometry and estimation
of nuclear genome size. Ann Bot – London 95: 99-110.
- Ducar E, Rewers M, Jedrzejczyk I, Martonfi P, Sliwinska E (2018).
Comparison of the genome size, endoreduplication and ISSR
markers polymorphism in eight
Lotus
(Fabaceae) species. Turk
J Bot 42: 1-14.
- Gajdošová A, Ostrolucká MG, Libiaková G, Ondrušková E, Šima D
(2006). Microclonal propagation of
Vaccinium
sp. and
Rubus
sp. and detection of genetic variability in culture in vitro.
Journal of Fruit and Ornamental Plant Research 14: 103-119.
- Galbraith DW, Harkins KR, Maddox JM, Ayres NM, Sharma DP,
Firoozabady E (1983). Rapid flow cytometric analysis of the
cell cycle in intact plant tissues. Science 220: 1049-1051.
- Gantait S, Mandal N, Bhattacharyya S, Das PK (2010). Sustainable
in vitro
propagation and clonal fidelity in strawberry.
International Journal of Plant Developmental
Biology 4: 19-25.
- Gantait S, Sinniah UR (2011). Morphology, flow cytometry and
molecular assessment of ex-vitro grown micropropagated
Anthurium
in comparison with seed germinated plants. Afr J
Biotechnol 10: 13991-13998.
- Hegnauer R (1990). Chemotaxonomie Der Pflanzen Band 9. Basel,
Switzerland: Birkhaüser, pp. 288-293.
- Huck CW, Huber CG, Ongania KH, Bonn GK (2000). Isolation
and characterization of methoxylated flavones in the
flowers of
Primula veris
by liquid chromatography and mass
spectrometry. J Chromatogr A 870: 453–462.
- ISTA (2015). International Rules for Seed Testing.
Vol. 2015, pp.
1-276.
- Kéry M, Matthies D, Spillmann HH (2000). Reduced fecundity and
offspring performance in small populations of the declining
grassland plants
Primula veris
and
Gentiana lutea
. J Ecol 88:
17-30.
- Lee JE, Safe S (2000). 3’,4’-Dimethoxyflavone as an aryl hydrocarbon
receptor antagonist in human breast cancer cells. Toxicol Sci
58: 235-242.
- Lehtilä K, Ehrlén J (2005). Seed size as an indicator of seed quality: a
case study of
Primula veris
. Acta Oecol 28: 207-212.
- Lehtilä K, Syrjänen K, Leimu R, Garcia MB, Ehrlén J (2005). Habitat
change and demography of
Primula veris
: identification and
management targets. Conserv Biol 29: 833-843.
- Marie D, Brown C (1993). A cytometric exercise in plant histograms,
with 2C values for 70 species. Biol Cell 78: 41-51.
- Matsumura Ch, Washitani I (2000). Effects of population size
and pollinator limitation on seed-set of
Primula sieboldii
populations in a fragmented landscape. Ecol Res 15: 307-322.
- Morozowska M (2002). Some factors affecting viability, dormancy
and germination of
Primula veris
(L.) seeds. Folia Horticulturae
14: 243-253.
- Morozowska M (2003). Flowering, fruiting and seed set of
Primula
veris
L. plants in field cultivation. Folia Horticulturae 2003/2:
80-82 (article in Polish with an abstract in English).
- Morozowska M (2004a). Studies on some seed traits of
Primula veris
L. plants from cultivated populations. Herba Pol 50: 137-144.
- Morozowska M (2004b). Vegetative development, flowering, fruiting
and saponin content in cultivated cowslip (
Primula veris
L.)
plants. Herba Pol 50: 28-35.
- Morozowska M, Czarna A, Kujawa M, Jagodziński AM (2011). Seed
morphology and endosperm structure of selected species of
Primulaceae, Myrsinaceae, and Theophrastaceae and their
systematic importance. Plant Syst Evol 291: 159-172.
- Morozowka M, Krzakowa M (2003). Genetic variation of
Primula
veris
in natural and cultivated populations. Acta Biol Cracov
Bot 45: 177-182.
- Morozowska M, Urbański Ł (2000). Characteristics of flowering
and fructifying and the evaluation of the seed production of
cowslip plants (
Primula veris
L.) from populations located on
the Wielkopolska Lowland. Roczniki Akademii Rolniczej w
Poznaniu Seria Botanika 3: 175-184 (article in Polish with an
abstract in English).
- Morozowska M, Wesołowska M (2004).
In vitro
clonal propagation
of
Primula veris
L. and preliminary phytochemical analysis.
Acta Biol Cracov Bot 46: 169-175.
- Price HJ, Hodnett G, Johnston JS (2000). Sunflower (
Helianthus
annuus
) leaves contain compounds that reduce nuclear
propidium iodide fluorescence. Ann Bot 86: 929-934.
- Rewers M, Jedrzejczyk I (2016). Genetic characterization of
Ocimum
genus using flow cytometry and inter-simple sequence repeat
markers. Ind Crop Prod 91: 142-151.
- Richards AJ (2002). Primula. 2nd ed. London, UK: BT Batsford.
- Shimono A, Washitani I (2007). Factors affecting variation in seed
production in the heterostylous herb
Primula modesta
. Plant
Spec Biol 22: 65-76.
- Siljak-Yakovlev S, Pustahija F, Solic EM, Bogunic F, Muratovic E,
Basic N, Catrice O, Brown SC (2010). Towards a genome size
and chromosome number database of Balkan flora: C-values
in 343 taxa with novel values for 242. Adv Sci Lett 3: 190-213.
- Sliwinska E, Thiem B (2007). Genome size stability in six medicinal
plant species propagated in vitro. Biol Plantarum 51: 556-558.
- Sliwinska E, Zielińska E, Jędrzejczyk I (2005). Are seeds suitable for
flow cytometric estimation of plant genome size? Cytom Part
A 64A: 72-79.
- Soltis DE, Soltis PS, Bennett MD, Leitch IJ (2003). Evolution of
genome size in the angiosperms. Am J Bot 90: 1596-1603.
- Stecher G, Huck CW, Stöggl WM, Bonn GK (2003). Phytoanalysis:
a challenge in phytomics. Trends in Analytical Chemistry 22:
1-14.
- Tamm CO (1972). Survival and flowering of perennial herbs. III. The
behavior of
Primula veris
on permanent plots. Oikos 23: 159-
166.
- Thiem B, Budzianowski J, Wesołowska M, Ratajczak L, Morozowska
M, Skrzypczak L (2008). Secondary metabolites of in vitro
cultures of selected Polish rare and endangered plants. Herba
Pol 54: 158-167.
- Thiem B, Sliwinska E (2003). Flow cytometric analysis of nuclear
DNA content in cloudberry (
Rubus chamaemorus
L.) in vitro
cultures. Plant Sci 164: 129-134.
- Tokalov SV, Kind B, Wollenweber E, Gutzeit HO (2004). Biological
effects of epicuticular flavonoids from
Primula denticulate
on
human leukaemia cells. J Agri Food Chem 52: 239-245.
- Tremayne A, Richards AJ (2000). Seed weight and seed number
affect subsequent fitness in outcrossing and selfing
Primula
species. New Phytol 148: 127-142.
- Wang KJ, Li XH, Yan MF (2014). Genetic differentiation in relation
to seed weights in wild soybean species (
Glycine soja
Sieb. &
Zucc.). Plant Syst Evol 300: 1729-1739.