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Effect of Ph on in Vitro Pollen Germination of Fourteen Cultivated and Wild Species of Cucurbit

M Rashed Zaman
Department of Genetic Engineering & Biotechnology, University of Rajshahi, Rajshahi-6205, Bangladesh

Abstract/ Executive Summary:

Context: The pH of the in vitro germination medium is the important factor controlling pollen germination and pollen tube development in different plant species. Objectives: The aim of the present study was to assess the effect of pH levels of germination medium on the germinability 14 species of cucurbit. Materials and Methods: In vitro pollen germination of 14 cucurbit species was examined at five different levels of pH using Brewbaker and Kwack’s medium. The pH of the medium was adjusted to 7.0, 7.5, 8.0, 8.5 and 9.0 by using a digital pH meter with the help of 0.1 N HCl and 0.1 N NaOH. In vitro pollen germination was tested using sitting drop culture technique. Five sitting drop cultures of pollen grains of five different pH for each species were raised on five different microslides. Then the slides were placed in a humidity chamber and incubated at 22 ± 2°C for 1 to 2 hrs. All the cultures were fixed by adding a drop of a 1-2% acetocarmine before start scoring and the germinated and non-germinated pollen grains were scored using a microscope. Results: The pollen germination rate was higher in increased pH level in most of the cucurbit species studied and at 9.0 pH the maximum germination was observed in pumpkin (98%), snake gourd (98%), ridge gourd (96%), pointed gourd (89%), cucumber (98%), bitter gourd (96%), sweet gourd (98%) and sponge gourd (93%). The highest percentage of germination was observed in bottle gourd (98%) cucumber short (96%), watermelon (98%), muskmelon (97%) and ivy gourd (87%) at 8.5 pH level. The only cucurbit species wax gourd showed highest germination (97%) at the lower pH level of 7.5. Analysis of variance for the 14 cucurbits at five different pH levels showed that the differences between the germination rates of pollen grains of 14 cucurbit species at different pH level were highly significant (F=12.49, P<0.001, at 13/52 df). Conclusion: In most of the investigated cucurbit species pH level 8.5 to 9.0 was found optimum for the highest number of in vitro pollen germination, while wax gourd responded maximum pollen germination at pH 7.5.

Keywords: Cucurbit, Pollen, Germination, pH.

Location:

Start Date:

End Date:

Program Area: Crop-Soil-Water Management

Commodity: Cucurbit

Objectives:

The aim of the present study was to assess the effect of pH levels of germination medium on the germinability 14 species of cucurbit.

Methodology:

The 14 cucurbit species were collected from natural population, commercial and homestead gardens used for the present work and they are pumpkin (Cucurbita maxima Duch ex Poir.), wax gourd (Benincasa hispida (Thunb.) Cogn.), bottle gourd (Lagenaria siceraria (Mol) Standl.), snake gourd (Trichosanthes cucumerina L.), ridge gourd (Luffa acutangula (Roxb.) L.), pointed gourd (Trichosanthes dioica Roxb.), cucumber (Cucumis sativus L. and Cucumis anguina L.), bitter gourd (Momordica charantia L.), sweet gourd (Momordica cochinchinensis Spreng.), watermelon (Citrullus lanatus Thunb.), muskmelon (Cucumis melo L.), sponge gourd (Luffa cylindrica Roem.) and ivy gourd (Coccinia cordifolia (Voigt) L.). Brewbaker and Kwack’s medium (Brewbaker and Kwack 1963) was used for the testing of effects of pH on in vitro pollen germination. The pH of the medium was adjusted to 7.0, 7.5, 8.0, 8.5 and 9.0 by using a digital pH meter with the help of 0.1 N HCl and 0.1 N NaOH. In vitro pollen germination was tested using sitting drop culture technique. Five sitting drop cultures of pollen grains of five different pH for each species were raised on five different microslides. To prevent the contamination of different medium with different pH, the needle was rinsed in distilled water each time after use. Then the slides were placed in a humidity chamber. A pair of petri plates with a moist filter paper or tissue paper lining the lower plate was served as an improvised humidity chamber. Two glass rods placed parallel at about 4 cm apart on the moist filter paper facilitated the humidity of the pollen germination. Then the humidity chambers containing cultures were incubated at 22 ± 2°C for 1 to 2 hrs. As the scoring of pollen germination takes considerable time, all the cultures were fixed by adding a drop of a 1-2% acetocarmine (i.e. fixative) before start scoring and the germinated and non-germinated pollen grains were scored using a microscope to calculate the percentage of pollen germination of the five different pH media.

Source of Information: J. bio-sci. 17: 129-133, 2009, ISSN 1023-8654

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Total Budget (Tk.) :

Achievement/Findings:

In most of the investigated cucurbit species pH level 8.5 to 9.0 was found optimum for the highest number of in vitro pollen germination, while wax gourd responded maximum pollen germination at pH 7.5.

Knowledge Management: Journal