Styrian oil pumpkin pollen germinability at higher irradiation doses: optimization of the in vitro germination protocol and irradiation procedure

Protocols for haploid induction in cucurbits are based on pollination with irradiated pollen, but the induction frequency is low and the majority of obtained embryos are zygotic. The longevity of Cucurbita pepo L. pollen is short even under natural conditions; following irradiation, germinability is decreased even further. This study was initiated to develop an optimal in vitro germination medium for styrian oil pumpkin (Cucurbita pepo L. subsp. pepo var. styriaca Greb.) pollen, which would enable accurate germination testing. Different pH values and the addition of sucrose, mannitol, and polyethylene glycol to the Brewbaker and Kwack germination medium were tested. The optimum medium condition was pH 9 and 12.5% (w/v) sucrose, while other tested components were not efficient. Using the optimized medium, X-ray-irradiated (100-700 Gy) pollen germinability was assessed under 2 air humidity conditions. Germinability of pollen irradiated at room humidity (RH) was generally lower than that of pollen irradiated at high humidity (HH). A major variability in pollen size (diameters ranged from 79.2 to 196.5 µm) and 2 subgroups were found in the pollen population. Following irradiation, HH conditions allowed germination of larger pollen grains than those of the nonirradiated control and RH.

Styrian oil pumpkin pollen germinability at higher irradiation doses: optimization of the in vitro germination protocol and irradiation procedure

Protocols for haploid induction in cucurbits are based on pollination with irradiated pollen, but the induction frequency is low and the majority of obtained embryos are zygotic. The longevity of Cucurbita pepo L. pollen is short even under natural conditions; following irradiation, germinability is decreased even further. This study was initiated to develop an optimal in vitro germination medium for styrian oil pumpkin (Cucurbita pepo L. subsp. pepo var. styriaca Greb.) pollen, which would enable accurate germination testing. Different pH values and the addition of sucrose, mannitol, and polyethylene glycol to the Brewbaker and Kwack germination medium were tested. The optimum medium condition was pH 9 and 12.5% (w/v) sucrose, while other tested components were not efficient. Using the optimized medium, X-ray-irradiated (100-700 Gy) pollen germinability was assessed under 2 air humidity conditions. Germinability of pollen irradiated at room humidity (RH) was generally lower than that of pollen irradiated at high humidity (HH). A major variability in pollen size (diameters ranged from 79.2 to 196.5 µm) and 2 subgroups were found in the pollen population. Following irradiation, HH conditions allowed germination of larger pollen grains than those of the nonirradiated control and RH.

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  • Agbagwa IO, Ndukwu BC, Mensah SI (2007). Floral biology, breeding system, and pollination ecology of Cucurbita moschata (Duch. ex Lam) Duch. ex Poir. varieties (Cucurbitaceae) from parts of the Niger Delta, Nigeria. Turk J Bot 31: 451–458.
  • Akbudak N, Seniz V (2009). In vitro and in vivo behavior of gamma- irradiated tomato (Lycopersicon esculentum) pollen. New Zeal J Crop Hort 37: 361–367.
  • Brewbaker JL, Emery GC (1962). Pollen radiobotany. Radiat Bot 1: 101–154.
  • Brewbaker JL, Kwack BH (1963). The essential role of calcium ion in pollen germination and pollen tube growth. Am J Bot 50: 859– 865.
  • Chalak L, Legave JM (1997). Effects of pollination by irradiated pollen in Hayward kiwifruit and spontaneous doubling of induced parthenogenetic trihaploids. Sci Hortic-Amsterdam 68: 83–93.
  • Conner PJ (2011). Optimization of in vitro pecan pollen germination. Hort Science 46: 571–576.
  • Cuny F, Grotte M, Dumas De Vaulx R, Rieu A (1993). Effects of gamma irradiation of pollen on parthenogenetic haploid production in muskmelon (Cucumis melo L.). Environ Exp Bot 33: 301–312.
  • Dafni A, Firmage D (2000). Pollen viability and longevity: practical, ecological and evolutionary implications. Plant Syst Evol 222: 113–132.
  • Denissen CJM, Den Nijs APM (1987). Effects of gamma irradiation on in vitro pollen germination of different Cucumis species. Euphytica 36: 651–658.
  • Franchi GG, Nepi M, Dafni A, Pacini E (2002). Partially hydrated pollen: taxonomic distribution, ecological and evolutionary significance. Plant Syst Evol 234: 211–227.
  • Gay G, Kerhoas C, Dumas C (1987). Quality of a stress-sensitive Cucurbita pepo L. pollen. Planta 171: 82–87.
  • Grouh MSH, Vahdati K, Lotfi M, Hassani D, Biranvand NP (2011). Production of haploids in Persian walnut through parthenogenesis induced by gamma-irradiated pollen. J Am Soc Hortic Sci 136: 198–204.
  • Hertwig O (1911). Die Radiumkrankheit tierischer Keimzellen. Arch Mikrosk Anat 77: 1–97 (article in German).
  • Košmrlj K, Murovec J, Bohanec B (2013). Haploid induction in hull- less seed pumpkin through parthenogenesis induced by X-ray- irradiated pollen. J Am Soc Hortic Sci 138: 310–316.
  • Kundu M, Dubey A, Srivastav M, Malik S, Singh B (2014). Effect of gamma ray irradiation and cryopreservation on pollen stainability, in vitro germination, and fruit set in Citrus. Turk J Biol 38: 1–9.
  • Kurtar ES (2009). Influence of gamma irradiation on pollen viability, germination ability, and fruit and seed set of pumpkin and winter squash. Afr J Biotechnol 8: 6918–6926.
  • Kurtar ES, Balkaya A (2010). Production of in vitro haploid plants from in situ induced haploid embryos in winter squash (Cucurbita maxima Duchesne ex Lam.) via irradiated pollen. Plant Cell Tiss Org 102: 267–277.
  • Kurtar ES, Balkaya A, Ozbakir M, Ofluoglu T (2009). Induction of haploid embryo and plant regeneration via irradiated pollen technique in pumpkin (Cucurbita moschata Duchesne ex. Poir). Afr J Biotechnol 8: 5944–5951.
  • Kurtar ES, Sari N, Abak K (2002). Obtention of haploid embryos and plants through irradiated pollen technique in squash (Cucurbita pepo L.). Euphytica 127: 335–344.
  • Murovec J, Bohanec B (2011). Haploids and doubled haploids in plant breeding. In: Abdurakhmonov IY, editor. Plant Breeding. Rijeka, Croatia: InTech, pp. 87–106.
  • Nepi M, Cresti L, Guarnieri M, Pacini E (2010). Effect of relative humidity on water content, viability and carbohydrate profile of Petunia hybrida and Cucurbita pepo pollen. Plant Syst Evol 284: 57–64.
  • Nepi M, Franchi GG, Pacini E (2001). Pollen hydration status at dispersal: cytophysiological features and strategies. Protoplasma 216: 171–180.
  • Pandey KK, Phung M (1982). ‘Hertwig effect’ in plants: induced parthenogenesis through the use of irradiated pollen. Theor Appl Genet 62: 295–300.
  • Rihova L, Hrabetova E, Tupy J (1996). Optimization of conditions for in vitro pollen germination and tube growth in potato. Int J Plant Sci 157: 561–566.
  • Sauton A, Dumas De Vaulx R (1987). Induction of gynogenetic haploid plants in muskmelon (Cucumis melo L.) by use of irradiated pollen. Agronomie 7: 141–148 (article in French with an abstract in English).
  • Speranza A, Calzoni GL, Pacini E (1997). Occurrence of mono- or disaccharides and polysaccharide reserves in mature pollen grains. Sex Plant Reprod 10: 110–115.
  • Sugiyama K, Morishita M (2000). Production of seedless watermelon using soft-X-irradiated pollen. Sci Hortic-Amsterdam 84: 255– 264.
  • Tejaswini (2002). Variability of pollen grain features: a plant strategy to maximize reproductive fitness in two species of Dianthus? Sex Plant Reprod 14: 347–353.
  • Vasil IK (1960). Studies on pollen germination of certain Cucurbitaceae. Am J Bot 47: 239–247.
  • Vižintin L, Bohanec B (2004). In vitro manipulation of cucumber (Cucumis sativus L.) pollen and microspores: isolation procedures, viability tests, germination, maturation. Acta Biol Cracov Bot 46: 177–183.
  • Yahata M, Yasuda K, Nagasawa K, Harusaki S, Komatsu H, Kunitake H (2010). Production of haploid plant of ‘Banpeiyu’ pummelo [Citrus maxima (Burm.) Merr.] by pollination with soft x-ray irradiated pollen. J Jpn Soc Hortic Sci 79: 239–245.
  • Zaman MR (2009). Effect of pH on in vitro pollen germination of fourteen cultivated and wild species of cucurbit. J Bio-Science 17: 129–133.
Turkish Journal of Biology-Cover
  • ISSN: 1300-0152
  • Yayın Aralığı: Yılda 6 Sayı
  • Yayıncı: TÜBİTAK
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Styrian oil pumpkin pollen germinability at higher irradiation doses: optimization of the in vitro germination protocol and irradiation procedure

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