Determining appropriate methods for estimating pollen viability and germination rates in lisianthus

Tuğba Kılıç

Department of Horticulture, Faculty of Agriculture, Yozgat Bozok University, Yozgat, Turkey
https://orcid.org/0000-0002-0528-7552

Ebru Sinanoğlu

Department of Horticulture, Faculty of Agriculture, Ankara University, Ankara, Turkey
https://orcid.org/0009-0008-4239-1333

Emine Kırbay

Atatürk Health Services Vocational School, Afyonkarahisar Health Sciences University, Afyonkarahisar, Turkey
https://orcid.org/0000-0002-0343-0829

Soner Kazaz

Department of Horticulture, Faculty of Agriculture, Ankara University, Ankara, Turkey
https://orcid.org/0000-0002-6644-9690

Sezai Ercişli

Department of Horticulture, Faculty of Agriculture, Atatürk University, Erzurum, Turkey
https://orcid.org/0000-0001-5006-5687


Abstract

Crossbreeding is a multi-stage process with inherent challenges and risks in developing new varieties. Success hinges on selecting highly fertile parents. In species like lisianthus, uncertainty persists regarding the optimal methods for assessing pollen quality, which is crucial for evaluating pollen parent fertility. This study seeks to identify the most reliable techniques for this purpose. Fresh and dead pollen from four lisianthus (Eustoma grandiflorum) varieties was used. The dead pollen was obtained by thermal inactivation. Five chemical staining methods (iodine-potassium iodide, 2,3,5-triphenyltetrazolium chloride – TTC, lactophenol cotton blue, safranin, acetocarmine) were employed to assess pollen viability, and two biological methods (Petri dishes, hanging drops) were used to determine the germination rate. Four solid medium cultures were employed in Petri dishes, while the hanging drop utilised four liquid medium cultures. Thirteen tests were conducted for each variety, evaluating fresh and dead pollen. The study found significant variations in pollen quality among lisianthus varieties and methods. Fresh pollen showed viability rates ranging from 56.87% to 99.41% and germination rates from 0.20% to 45.11%. TTC exhibited the lowest viability rate across all varieties, while the highest germination rate was observed in the liquid culture medium with only boric acid and PEG1500. Notably, TTC was the sole viability method that did not stain dead pollen, and no germination occurred in any method for dead pollen. TTC is the most reliable staining method, and a liquid culture medium with boric acid and PEG1500 effectively determines lisianthus pollen quality. Varying boric acid and PEG1500 concentrations are advisable.

Keywords:

antioxidant, antimicrobial, chemical composition, Diospyros lotus, macro and micronutrient

Abdelgadir, H.A., Johonson, S.D., Van Staden, J. (2012). Pollen viability, pollen germination and pollen tube growth in the biofuel seed crop Jatropha curcas (Euphorbiaceae). S. Afr. J. Bot., 79, 132–139. https://doi.org/10.1016/j.sajb.2011.10.005 DOI: https://doi.org/10.1016/j.sajb.2011.10.005

Bolat, İ., Güleryüz, B. (1994). Bazı kayısı çeşitlerinde çiçek tozu canlılık ve çimlenme düzeyleri ile bunlar arasındaki ilişkinin belirlenmesi üzerine bir araştırma [A study on the determination of pollen viability and germination and their interaction of some apricot cultivars]. Atatürk Üniversitesi Ziraat Fakültesi Dergisi, 25(3), 344–353.

Bolat, İ., Pırlak, L. (1999). An investigation on pollen viability, germination and tube growth in some stone fruits. Turk. J. Agric. Forest., 23, 383–388.

Erbaş, S, Alagöz, M, Baydar, H. (2015). Research on flower morphology and pollen viability of oil-bearing rose (Rosa damascena Mill.). J. Fac. Agric. 10(2), 40–50.

Eti, S. (1990). Çiçek tozu miktarını belirlemede kullanılan pratik bir yöntem [A practical method for the determination of pollen production]. Çukurova Üniv. Ziraat Fakültesi Der. 5(4), 49–58.

Eti, S. (1991). Determination of pollen viability and germination capability of some fruit species and cultivars by different in vitro tests. Çukurova Univ. J. Agric. Fac., 6, 69–80.

Farooq, A., Khan, M.A., Riaz, A., Younis, A., Butt, S.J., Nadeem, M. (2013). Compatibility evaluation of various scented rosa species through cross pollination. Int. J. Basic Appl. Sci. 13(2), 21–27.

Farooq, A., Lei, S., Nadeem, M., Asif, A., Akhtar, G., Butt, S.Ş. (2016). Cross compatibility in various scented Rosa species breeding. Pakistan J. Agric. Sci., 53(4), 863–869. DOI: https://doi.org/10.21162/PAKJAS/16.1817

Fragallah, S.A.D.A., Lin, S., Li, N., Ligat, E.J., Chen, Y. (2019). Effects of sucrose, boric acid, pH, and incubation time on in vitro germination of pollen and tube growth of chinese fir (Cunnighamial lanceolata L.). Forests, 10(2), 102. https://doi.org/10.3390/f10020102 DOI: https://doi.org/10.3390/f10020102

Giovannini, A., Macovei, A., Donà, M., Valassi, A., Caser, M., Mansuino, A., Balestrazzi, A. (2013). Pollen grain preservation at low temperatures in valuable commercial rose cultivars. In: VI International Symposium on Rose Research and Cultivation, 1064, 63–66. DOI: https://doi.org/10.17660/ActaHortic.2015.1064.7

Hong-Qi, Z., Croes, A.F. (1982). A new medium for pollen germination in vitro. Acta Bot. Neerl., 31(1/2), 113–119. DOI: https://doi.org/10.1111/j.1438-8677.1982.tb01597.x

Huang, J., Zhao, X., Cheng, K., Jiang, Y., Ouyang, Y., Xu, C., Li, X., Xiao, J., Zhang, Q. (2013). OsAP65, a rice aspartic protease, is essential for male fertility and plays a role in pollen germination and pollen tube growth. J. Exp. Bot., 64(11), 3351–3360. https://doi.org/10.1093/jxb/ert173 DOI: https://doi.org/10.1093/jxb/ert173

İmrak, B. (2010). Bazı kiraz çeşitlerinin subtropik iklim koşullarındaki performansları ve çoklu dişi organ oluşumu sorununun çözümüne ilişkin araştırmalar [Performances of some sweet cherry cultivars (Prunus avium L.) under the subtropical climatic conditions and researches to the solution of multiple pistil]. Çukurova Üniversitesi, Fen Bilimleri Enstitüsü, Doktora Tezi. 193.

Jayaprakash, P. (2018). Pollen germination in vitro. In: Pollination in plants, Mokwala P.W. (ed.). Intech Open, 81, 81–96.

Jiang, Y., Bueckert, R.A., Warkentin, T.D., Davis, A.R. (2017). High temperature effects on in vitro pollen germination and seed set in field pea. Canad. J. Plant Sci., 98, 71–80. https://doi.org/10.1139/cjps-2017-0073 DOI: https://doi.org/10.1139/CJPS-2017-0073

Kalyoncu, İ.H., Ersoy, N., Yılmaz, M. (2013). Determination of pollen viability germination and production of selected K-3 cornellian cherry (Cornus mas L.) type. Manas J. Agric. Life Sci., 3(1), 39–45.

Li, M., Jiang, F., Huang, L., Wang, H., Song, W., Zhang, X., Niu, L. (2023). Optimization of in vitro germination, viability tests and storage of paeonia ostii pollen. Plants, 12(13), 2460. https://doi.org/10.3390/plants12132460 DOI: https://doi.org/10.3390/plants12132460

Lin, Y., Wang, Y., Iqbal, A., Shi, P., Li, J., Yang, Y., Lei, X. (2017). Optimization of culture medium and temperature for the in vitro germination of oil palm pollen. Sci. Hortic., 220, 134–138. DOI: https://doi.org/10.1016/j.scienta.2017.03.040

Macovei, A., Caser, M., Mattia, D.O.N.À., Valassi, A., Giovannini, A., Carbonera, D., Balestrazzi, A. (2016). Prolonged cold storage affects pollen viability and germination along with hydrogen peroxide and nitric oxide content in Rosa hybrida. Not. Bot. Horti Agrobot. Cluj-Napoca, 44(1), 6–10. DOI: https://doi.org/10.15835/nbha44110357

Martins, E.S., Davide, L.M.C., Miranda, G.J., Barizon, J.D.O., Souza, F.D.A., Carvalho, R.P.D., Gonçalves, M.C. (2016). In vitro pollen viability of maize cultivars at different times of collection. Ciência Rural, 47(2), e20151077. DOI: https://doi.org/10.1590/0103-8478cr20151077

Melloni, M.L.G., Scarpari, M.S., Mendonça, J.R., Perecin, D., Landell, M.G.A., Pinto, L.R. (2013). Comparison of two staining methods for pollen viability studies in Sugarcane. Sugar Tech., 15(1), 103–107. https://doi.org/10.1007/s12355-012-0185-6 DOI: https://doi.org/10.1007/s12355-012-0185-6

Nadeem, M., Akond, M., Riaz, A., Qasim, M., Younis, A., Farooq, A. (2013). Pollen morphology and viability relates to seed production in hybrid roses. Plant Breed. Seed Sci., 68, 25–38. DOI: https://doi.org/10.2478/v10129-011-0078-y

Pacini, E., Dolferus, R. (2019). Pollen developmental arrest: maintaining pollen fertility in a world with a changing climate. Front. Plant Sci., 10, 679. https://doi.org/10.3389/fpls.2019.00679 DOI: https://doi.org/10.3389/fpls.2019.00679

Parfitt, D.E., Ganeshan, S. (1989). Comparison of procedures for estimating viabilitiy of Prunus polen. HortScience, 24 (2), 354–356. DOI: https://doi.org/10.21273/HORTSCI.24.2.354

Paydaş, S., Eti, S., Kaşka, N., Sayılıkan, G. (1996). Pozantı ekolojik koşullarında yetiştirilen bazı kiraz çeşitlerinde çiçek tozu canlılık ve çimlenme yetenekleri ile üretim miktarlarının belirlenmesi [Determination of flower powder viability, germination ability and production amount of some cherry cultivars grown in Pozantı ecological conditions.]. Çukurova Üniversitesi Ziraat Fakültesi Dergisi, 11(3), 149–160.

Rodriguez-Enriquez, M.J., Mehdi, S., Dickinson, H.G., Grant-Downton, R.T. (2013). A novel method for efficient in vitro germination and tube growth of Arabidopsis thaliana pollen. New Phyt., 197, 668–679. DOI: https://doi.org/10.1111/nph.12037

Royal Flora Holland, (2023). Annual reports 2022. https://www.royalfloraholland.com/en/press-releases/annual-report-2022 [date of access: 21.05.2023].

Saarela, J.M. (2012). Taxonomic synopsis of invasive and native Spartina (Poaceae, Chloridoideae) in the Pacific Northwest (British Columbia, Washington and Oregon), including the first report of Spartina × townsendii for British Columbia, Canada. PhytoKeys, 10(10), 25–82. DOI: https://doi.org/10.3897/phytokeys.10.2734

Sanşili, Ç. (2014). Bazı elma ve kiraz çeşitlerinde çiçek tozu performanslarının belirlenmesi [Determination of pollen performances of some apple and cherry cultivars]. Yüksek Lisans Tezi, Namık Kemal Üniversitesi, Fen Bilimleri Enstitüsü, Bahçe Bitkileri Anabilim Dalı, 70, Tekirdağ.

Şensoy, A.S., Ercan, N., Ayar, F., Temirkaynak, M. (2003). Cucurbitaceae familyasındaki bazı sebze türlerinde çiçek tozlarının bazı morfolojik özellikleri ile canlılıklarının belirlenmesi [Determination of pollen performances of some apple and cherry cultivars]. Akdeniz Univ. J. Fac. Agric., 16(1), 1–6.

Shivanna, K.R., Sawhney, V.K. (1995). Polyethylene glycol improves the in vitro growth of Brassica pollen tubes without loss in germination. J. Experim. Bot., 46(11),1771–1774. DOI: https://doi.org/10.1093/jxb/46.11.1771

Soares, T.L., Jesus, O.N.D., Santos-Serejo, J.A.D., Oliveira, E.J.D. (2013). In vitro pollen germination and pollen viability in passion fruit (Passiflora spp.). Rev. Bras. Frutic., 35, 1116–1126. DOI: https://doi.org/10.1590/S0100-29452013000400023

Stanley, R.G., Linskens H.F. (1974). Pollen: biology, biochemistry and management. NewYork: Berlin-Heidelberg. DOI: https://doi.org/10.1007/978-3-642-65905-8

Tingting, X. (2009). Measurement and marker trait association analysis of pollen viability in the K5 tetraploid rose population. Thesis, Department of Plant Breeding Wageningen University.

Tushabe, D., Rosbakh, S. (2021). A compendium of in vitro germination media for pollen research. Front. Plant Sci., 12, 709945. DOI: https://doi.org/10.3389/fpls.2021.709945

Velasco-Ramírez, A., Villegas-Lozano, M., Velasco-Ramírez, A.P., Hernández-Herrera, R.M., Hernández-Pérez, A.R., García-Sahagún, M.L., Torres-Morán, M.I. (2022). Effect of daminozide on the growth and flowering of Eustoma grandiflorum propagated in potted. Acta Sci. Pol. Hortorum Cultus, 21(3), 75–82. http://dx.doi.org/10.24326/asphc.2022.3.7 DOI: https://doi.org/10.24326/asphc.2022.3.7

Wang, Z-Y, Ge, Y., Scott, M., Spangenberg, G. (2004). Viability and longevity of pollen from transgenic and nontransgenic tall fescue (festuca arundinacea) (poaceae) plants. Am. J. Bot., 91(4), 523–530. DOI: https://doi.org/10.3732/ajb.91.4.523

Wang, Q., Zhang, Y., Kawabata, S., Li, Y. (2011). Double fertilization and embryogenesis of Eustoma grandiflorum. J. Japan. Soc. Hortic. Sci., 80(3), 351–357. DOI: https://doi.org/10.2503/jjshs1.80.351

Wani, T.A., Pandith, S.A., Rana, S., Bhat, W.W., Dhar, N., Razdan, S., Chandra, S., Kitchlu, S., Sharma, N., Lattoo, S.K. (2015). Promiscuous breeding behaviour in relation to reproductive successin Grewia asiatica L. (Malvaceae). Flora, 62–71. DOI: https://doi.org/10.1016/j.flora.2014.11.002

Zhao, H., Chen, F., Fang, W. (2005). Pollen germination in vitro of chrysanthemum cultivars with small inflorescences and several species of Dendranthema. J. Nanjing Agric. Univ., 28(2), 22–27.

Zhao, H., Chen, F., Wang, Y., Chen, S., Fang, W., Guo, W. (2008). Study on pollen viability, longevity, and pistil receptivity of self-compatible Chrysanthemum with small inflorescences. XXVII International Horticultural Congress-IHC2006: International Symposium on Ornamentals, ISHS Acta Horticulturae 766, Korea (Seoul), March 31, 402–415.

Zlesak, D.C. (2007). Rose. Rosa × hybrida. In: Flower breeding and genetics. Issues, challenges and opportunities for the 21st century. Dordrecht, Springer Netherlands, 695–740. DOI: https://doi.org/10.1007/978-1-4020-4428-1_26

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Published
2024-06-28



Tuğba Kılıç 
Department of Horticulture, Faculty of Agriculture, Yozgat Bozok University, Yozgat, Turkey https://orcid.org/0000-0002-0528-7552
Ebru Sinanoğlu 
Department of Horticulture, Faculty of Agriculture, Ankara University, Ankara, Turkey https://orcid.org/0009-0008-4239-1333
Emine Kırbay 
Atatürk Health Services Vocational School, Afyonkarahisar Health Sciences University, Afyonkarahisar, Turkey https://orcid.org/0000-0002-0343-0829
Soner Kazaz 
Department of Horticulture, Faculty of Agriculture, Ankara University, Ankara, Turkey https://orcid.org/0000-0002-6644-9690
Sezai Ercişli 
Department of Horticulture, Faculty of Agriculture, Atatürk University, Erzurum, Turkey https://orcid.org/0000-0001-5006-5687



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