Optimising storage and in vitro germination of Eucalyptus pollen

2007 ◽  
Vol 55 (1) ◽  
pp. 83 ◽  
Author(s):  
Tasmien N. Horsley ◽  
Steven D. Johnson ◽  
Terrence K. Stanger

The best sucrose solution for maximum in vitro germination of Eucalyptus pollen was investigated in order to evaluate pollen germination rate as an indicator of pollen viability. In vitro germination of both freshly collected and 1-year-old pollen (stored at 4°C) of Eucalyptus grandis, E. smithii, E. nitens, E. dunnii and E. macarthurii was carried out in 0, 10, 20, 30, 40 and 50% (w/v) sucrose solutions, either with (0.15 mg L–1) or without boric acid. Similar trends were obtained for both fresh and 1-year-old pollen, with all species responding most favourably to 30% (w/v) sucrose and 0.15 mg L–1 boric acid. When an optimal in vitro germination medium had been established, the viabilities (%germination) of E. smithii, E. nitens and E. grandis pollen, stored at room (25°C), fridge (4°C), freezer (–10°C) and liquid nitrogen (–196°C) temperatures, were compared. For all tested species, germination declined as storage temperature increased, and by 8 months, the highest survival was obtained with cryostored pollen.

2007 ◽  
Vol 21 (1) ◽  
pp. 23-29 ◽  
Author(s):  
Ian C. Burke ◽  
John W. Wilcut ◽  
Nina S. Allen

A high proportion of viable pollen grains must germinate to study the physiology of pollen growth to reduce the confounding effects of environmental influences on pollen germination. The objectives of this study were to evaluate the nuclear state and develop a suitable medium and culture method for in vitro germination of johnsongrass pollen. Johnsongrass pollen was trinucleate, and in vitro tests for pollen viability using Alexander's stain and a fluorochromatic reaction method (FCR) indicated johnsongrass pollen was viable (92.6 to 98.4%). A factorial treatment arrangement of four concentrations of sucrose, two concentrations of boric acid, and two concentrations of calcium nitrate were used to determine the optimum pollen-germination medium composition in suspension culture, agar culture, and cellophane membrane culture. Germination was highest in a suspension culture with a medium containing 0.3 M sucrose, 2.4 mM boric acid, and 3 mM calcium nitrate. Pollen germination using this medium was 78.9% when anthers were harvested just before anthesis.


Forests ◽  
2019 ◽  
Vol 10 (2) ◽  
pp. 102 ◽  
Author(s):  
Seif Fragallah ◽  
Sizu Lin ◽  
Nuo Li ◽  
Elly Ligate ◽  
Yu Chen

In vitro pollen germination provides a novel approach and strategy to accelerate genetic improvement of tree breeding. Studies about pollen germination and tube growth of Chinese fir are limited. Therefore, this study aimed to investigate the effects of sucrose, boric acid, pH, and time of incubation on pollen germination and tube growth. Pollen from 9 clones were selected. In vitro germination was performed in basic media as control, and in different concentrations of sucrose (0, 10 and 15%), boric acid (0.01, 0.1 and 0.2%), and pH levels (4.5, 5 and 7). Pollen germination rates and tube growth were recorded periodically at 1, 12, 24, and 48 h. The results showed that sucrose imposes significant effects on pollen germination and tube growth. The effects are most obvious at concentration of 15%. Boric acid significantly promoted germination and tube growth. The promotion was most notable in lower concentration of 0.01%. The media adjusted to pH 7.0 boosted the germination and pollen tube growth. The optimum time of incubation was 24 and 48 h for pollen germination and tube growth, respectively. Sucrose, pH, and time of incubation were positively correlated, whereas boric acid negatively correlated with pollen germination and tube growth. This study provided experimental evidences for selecting viable pollens for Chinese fir breeding.


1992 ◽  
Vol 47 (1-2) ◽  
pp. 102-108 ◽  
Author(s):  
J. Polster ◽  
M. Schwenk ◽  
E. Bengsch

Abstract It is possible to obtain pollen germination and pollen tube growth in vitro if boric acid is present. In this work the effect was studied using as a semiquantitative parameter the mean length (l̄) of Lilium longiflorum pollen tubes. Pollen tube growth was examinated in dependence on boric acid, ortho-silicic acid, nucleic bases, Ca2+ and Zn2+ in 10% sucrose solution. The maximum of l̄ is obtained for concentrations between 2-20 ppm boron. The simultaneous supply of silicon added as water glass leads to a synergistic stimulation effect on pollen tube growth and facilitates branching. The silicon action is preceded of a pollen tube growth inhibition period during 3 h. Adenine and guanosine are able to substitute partially boron as pollen germination and pollen tube growth stimulator. Concentrations of 100 ppm adenine leads to half the boron effect. The same stimulation effect is obtained by guanosine. Ca2+ can partially substitute boron as well. The stimulation action of boron is significantly attenuated by Zn2+ and by the herbicide Dicuran. These and preceding results from physiological studies indicate that boron and silicon should be essential trace elements for the regulation of molecular biological processes.


HortScience ◽  
2004 ◽  
Vol 39 (2) ◽  
pp. 365-367 ◽  
Author(s):  
Zhanao Deng ◽  
Brent K. Harbaugh

The sporadic nature of inflorescence production and flower protogyny in caladium (Caladium ×hortulanum Birdsey) makes it desirable to store pollen and to rapidly assess its viability for cross-pollinations in breeding programs. This study was conducted to develop a procedure to determine caladium pollen viability and to use that procedure to evaluate the effect of short-term storage conditions on pollen viability. The sucrose level in the culture medium was found to have a significant impact on the in vitro germination of caladium pollen; a concentration of 6.8% was determined to be optimal for pollen germination. Caladium pollen lost viability within 1 day under room (24 °C) or freezing (-20 °C) temperatures, but could be stored at 4 °C for 2 to 4 days. Pollen stored at 4 °C produced successful pollinations. Data obtained from large-scale greenhouse pollinations supported use of this in vitro germination assay as a convenient way to evaluate caladium pollen viability (and fertility).


2017 ◽  
Vol 32 (1) ◽  
pp. 61-65 ◽  
Author(s):  
Natalia Georgieva ◽  
Ivelina Nikolova ◽  
Valentin Kosev ◽  
Yordanka Naydenova

The objective of this study was to evaluate the influence of two organic nanofertilizers, Lithovit and Nagro, on in vitro germination, pollen tube elongation and pollen grain viability of Pisum sativum L cv. Pleven 4. The effect of their application was high and exceeded data for the untreated control (44.2 and 47.23 % regarding pollen germination and pollen tube elongation, respectively), as well as the effect of the control organic algal fertilizer Biofa (17.5 and 27.9 %, respectively). Pollen grains were inoculated in four culture media. A medium containing 15% sucrose and 1% agar had the most stimulating impact on pea pollen grains. Pollen viability, evaluated by staining with 1% carmine, was within limits of 74.72-87.97%. The highest viability of pollen grains was demonstrated after the application of Nagro organic nano-fertlizer.


2013 ◽  
Vol 35 (4) ◽  
pp. 1116-1126 ◽  
Author(s):  
Taliane Leila Soares ◽  
Onildo Nunes de Jesus ◽  
Janay Almeida dos Santos-Serejo ◽  
Eder Jorge de Oliveira

The use of Passiflora species for ornamental purposes has been recently developed, but little is known about pollen viability and the potential for crossing different species. The objective of this study was to evaluate the pollen viability of six Passiflora species collected from different physiological stages of development through in vitro germination and histochemical analysis using dyes. The pollen was collected in three stages (pre-anthesis, anthesis and post-anthesis). Three compositions of culture medium were used to evaluate the in vitro germination, and two dyes (2,3,5-triphenyltetrazolium chloride, or TTC, and Lugol's solution) were used for the histochemical analysis. The culture medium containing 0.03% Ca(NO3) 4H2O, 0.02% of Mg(SO4 ).7H2O, 0.01% of KNO3, 0,01% of H3BO3, 15% sucrose, and 0.8% agar, pH 7.0, showed a higher percentage of pollen grains germinated. Anthesis is the best time to collect pollen because it promotes high viability and germination. The Lugol's solution and TTC dye overestimated the viability of pollen, as all accessions showed high viability indices when compared with the results obtained in vitro.


1998 ◽  
Vol 123 (1) ◽  
pp. 41-46 ◽  
Author(s):  
Raphael A. Stern ◽  
Shmuel Gazit

The lychee (Litchi chinensis Sonn.) has two types of pollen-releasing flowers—M1 and M2. We compared the morphology and viability of these two pollen types, mainly for the two commercial cultivars in Israel: `Mauritius' and `Floridian'. Observation by scanning electron microscope did not reveal any consistent morphological differences between the two pollen types. However, M2 pollen was found to have a consistent and significant advantage over M1 pollen in in vitro germination tests. M2 pollen from `Mauritius', `Floridian', `No Mai Chee', `Wai Chee', and `Early Large Red' had a much higher germination rate at 15, 20, 25, 30, and 35 °C than M1 pollen from those same cultivars. The optimal incubation temperature for in vitro pollen germination was 30 °C for M2 pollen of all five cultivars studied; adequate germination rates were also found at 35 and 25 °C. The optimal temperature for M1 pollen germination was also 30 °C for `Mauritius' and `No Mai Chee', but was not well defined for the other three cultivars. No pronounced advantage of M2 pollen-tube growth could be discerned 48 h after hand pollination. However, final fruit set was consistently and significantly higher after hand pollination with M2 pollen, relative to M1 pollen. Hot (32/27 °C) and warm (27/22 °C) regimes during flower development had a pronounced detrimental effect on pollen viability compared to a cool (22/17 °C) regime. `Floridian' was much more susceptible than `Mauritius' in this respect.


HortScience ◽  
1997 ◽  
Vol 32 (5) ◽  
pp. 916-917 ◽  
Author(s):  
Tina L. Maguire ◽  
Margaret Sedgley

Storage of Banksia menziesii pollen was assessed at 20, 4, –20, –80, and –196 °C using a semisolid medium of 1% agar, 15% sucrose, 0.01% boric acid, 0.03% calcium nitrate, 0.02% magnesium sulfate, 0.01% potassium nitrate, and an incubation temperature of 25 °C. The germination percentage remained constant at ≈70% in all treatments except for pollen stored at 20 °C, which exhibited only 25% germination after 6 months. Pollen viability was assessed using fluorescein diacetate (FDA), but the results did not reflect the loss of germinability at 20 °C and correlation with in vitro results was variable. Floret position on the inflorescence did not affect germination, but pollen viability varied over the flowering period with maximum germination at midseason.


2021 ◽  
Vol 25 (04) ◽  
pp. 838-844
Author(s):  
Wenqing Jia

Knowledge about pollen ultra-morphology, storage characteristics and germination rate are essential for directional plant breeding and plant improvement. The objective of this study was to determine a suitable medium for pollen germination in vitro of Viola dissecta and to evaluate the effect of different storage temperatures on its pollen longevity. The pollen ultra-morphology of V. dissecta was observed using scanning electron microscopy (SEM), the suitable medium for pollen germination in vitro was determined by orthogonal test. Dried pollen of V. dissecta was stored at different temperatures (room temperature, 4, -20 and -80°C) and different storage times (24, 40, 72, 120, 184, 264 and 365 d), the germination rate of the stored pollen and the activities of SOD, POD and CAT were investigated. Pollen grains of V. dissecta were medium-sized with three germination ditches. The surface ornamentation was smooth with small grains set on the surface, which was different from Viola spp. pollen. The most suitable medium for V. dissecta was composed of 285 g•L-1 sucrose, 6 g•L-1 agar, 50 mg•L-1 GA3, 250 mg•L-1 boric acid, and 200 mg•L-1 Ca(NO3)2, The best storage temperature of pollen was -80oC, after 365 d of storage, the germination rate was still 57.86%. During storage, the pollen germination rate decreased significantly after the peak of the activities of the three antioxidant enzymes. Correlation analysis showed that SOD was major factor affecting the germination rate of V. dissecta pollen, and it has a significant positive correlation with pollen germination rate, followed by CAT and POD. SOD was a sensitive antioxidant enzyme at room temperature, 4 and -80°C, whereas at -20°C, both SOD and CAT were sensitive antioxidant enzymes. © 2021 Friends Science Publishers© 2021 Friends Science Publishers© 2021 Friends Science Publishers© 2021 Friends Science Publishers© 2021 Friends Science Publishers© 2021 Friends Science Publishers© 2021 Friends Science Publishers© 2021 Friends Science Publishers© 2021 Friends Science Publishers© 2021 Friends Science Publishers© 2021 Friends Science Publishers© 2021 Friends Science Publishers© 2021 Friends Science Publishers© 2021 Friends Science Publishers© 2021 Friends Science Publishers© 2021 Friends Science Publishers© 2021 Friends Science Publishers© 2021 Friends Science Publishers© 2021 Friends Science Publishers© 2021 Friends Science Publishers© 2021 Friends Science Publishers© 2021 Friends Science Publishers© 2021 Friends Science Publishers© 2021 Friends Science Publishers © 2021 Friends Science Publishers© 2021 Friends Science Publishers© 2021 Friends Science Publishers© 2021 Friends Science Publishers© 2021 Friends Science Publishers© 2021 Friends Science Publishers© 2021 Friends Science Publishers©


2014 ◽  
Vol 3 (3) ◽  
pp. 146-153
Author(s):  
Reshmi Chatterjee ◽  
Satadip Sarkar ◽  
GM Narasimha Rao

Pollen germination forms one of the most important stage post pollination prior to fertilization. This is essential for proper seed setting and seed development. In vitro pollen germination test is the most reliable way of assessing the pollen viability. In the present study pollen grains of seven genera under Apocynaceae family namely, Allamanda, Alstonia, Catharanthus, Nerium, Plumeria, Thevetia and Tabernaemontana were tested in some basic cultural media, such as Brewbaker’s media, 6% Glucose solution, 4% Calcium Nitrate solution and 3% Boron solution. Alstonia pollen grains exhibited highest percentage of germination rate in all the cultural media. Glucose and Brewbaker’s media is found to be highly suitable for efficient pollen germination in all the genera. Boron solution is effective for germination of pollen grains of tree species. In vitro pollen germination can be easily carried out in laboratories. These results can be utilised in plant breeding programmes to improve cultivar and varieties. DOI: http://dx.doi.org/10.3126/ije.v3i3.11074 International Journal of Environment Vol.3(3) 2014: 146-153


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