scholarly journals 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 ◽  
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.

1989 ◽  
Vol 37 (5) ◽  
pp. 429 ◽  
Author(s):  
BM Potts ◽  
JB Marsden-Smedley

The effect of boric acid (0-450 ppm) and sucrose (0-40%) on pollen germination and pollen tube growth in Eucalyptus globulus, E. morrisbyi, E. ovata and E. tirnigera was examined in vitro. Over the con- centrations tested, sucrose had by far the largest effect upon both pollen germination and tube lengths. The optimum sucrose concentration for pollen germination (30%) and pollen tube growth (20%) differed markedly with very little (<lo%) germination occurring in the absence of sucrose. The interaction of sucrose and boric acid was significant. However, in general both pollen germination and pollen tube growth were increased by the addition of up to 100 ppm boric acid, but above this level the response plateauxed. The four species differed significantly in their pattern of response to both boric acid and sucrose and the predicted optima derived from analysis of response surfaces differed between species. The predicted sucrose concentration for optimal germination and growth of E. urnigera pollen was consistently less than the other species and in terms of the optimal level of boric acid for pollen tube growth species can be ranked in the order E. globulus > E. ovata > E. morrisbyi = E. urnigera. Pollen germination and tube growth of all four species on a medium comprising 20% sucrose and 200 ppm boric acid would not differ significantly from the observed maximum response of each species and this could suffice as a generalised medium. However, if only percentage germination is to be assessed 30% sucrose would be preferable. It is argued that subtle interspecific differences in optimal in vitro con- ditions for pollen germination and pollen tube growth are likely to reflect differences in pollen physiology which in vivo may have important implications for the success of hybridisation where pollen competition occurs.


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.


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.


Molecules ◽  
2018 ◽  
Vol 23 (12) ◽  
pp. 3162 ◽  
Author(s):  
Seif Aldin Dawina Abdallah Fragallah ◽  
Pei Wang ◽  
Nuo Li ◽  
Yu Chen ◽  
Sizu Lin

Pollen grains produce certain metabolites, which can improve or inhibit germination and tube growth. Metabolomic analysis of germinating and growing Chinese fir pollen has not been reported. Therefore, this study aimed to analyse metabolites changes, content and expression in the germinating pollen of Chinese fir. To understand the metabolic differences, two clones from Chinese fir were selected. Metabolomics analyses were performed on three stages (1-, 24- and 48-h) during in vitro pollen germination. The metabolites profiles at different time points were analyzed by using liquid chromatography-mass spectrometry. The results showed that 171 peaks were screened; the corresponding differential metabolites of 121 peaks were classified into nine types of substances. The expression of metabolites showed significant differences across and between clones, and the variation was evident at all germination stages. The expression was obvious at the early stage of germination, which differed clearly from that of the late stage after pollen tube growth. Moreover, the metabolites were mainly enriched in 14 metabolic pathways. Pollen germination and tube growth and metabolites expressions changed per incubation time. Since this work is preliminary, we suggest further investigations to understand the relationship between the differential metabolites and pollen development, and factors affecting pollen germination process.


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.


2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Biying Dong ◽  
Qing Yang ◽  
Zhihua Song ◽  
Lili Niu ◽  
Hongyan Cao ◽  
...  

AbstractMature pollen germinates rapidly on the stigma, extending its pollen tube to deliver sperm cells to the ovule for fertilization. The success of this process is an important factor that limits output. The flavonoid content increased significantly during pollen germination and pollen tube growth, which suggests it may play an important role in these processes. However, the specific mechanism of this involvement has been little researched. Our previous research found that hyperoside can prolong the flowering period of Abelmoschus esculentus (okra), but its specific mechanism is still unclear. Therefore, in this study, we focused on the effect of hyperoside in regulating the actin-depolymerizing factor (ADF), which further affects the germination and growth of pollen. We found that hyperoside can prolong the effective pollination period of okra by 2–3-fold and promote the growth of pollen tubes in the style. Then, we used Nicotiana benthamiana cells as a research system and found that hyperoside accelerates the depolymerization of intercellular microfilaments. Hyperoside can promote pollen germination and pollen tube elongation in vitro. Moreover, AeADF1 was identified out of all AeADF genes as being highly expressed in pollen tubes in response to hyperoside. In addition, hyperoside promoted AeADF1-mediated microfilament dissipation according to microfilament severing experiments in vitro. In the pollen tube, the gene expression of AeADF1 was reduced to 1/5 by oligonucleotide transfection. The decrease in the expression level of AeADF1 partially reduced the promoting effect of hyperoside on pollen germination and pollen tube growth. This research provides new research directions for flavonoids in reproductive development.


2021 ◽  
Vol 74 ◽  
Author(s):  
Thomas Sawidis ◽  
Gülriz Baycu ◽  
Elżbieta Weryszko-Chmielewska ◽  
Aneta Sulborska

Abstract In vitro culture of Lilium longiflorum pollen grains was carried out to determine the role of manganese in pollen germination and pollen tube growth. Pollen germination was adversely affected by the presence of manganese (>10 −8 M), whereas low concentrations (10 −12 –10 −10 M) stimulated the process. Manganese caused morphological anomalies during tube growth, characterized by irregular pollen tube thickening and swollen tips. The main effect was the anomalous cell wall formation at the tip, in which the presence of several organelles reduced the number of secretory vesicles. A loose network of fibrillar material and spherical aggregates, mostly in the tip region, was detected, and this material was progressively loosened into the surrounding medium. As a response to potential toxicity, the excess manganese was isolated in vacuoles, which formed an internal barrier against penetration of manganese to the tip area. Elevated manganese concentrations might affect plant reproduction, resulting in anomalies in gamete development. Consequently, the loss in genetic diversity and decreased fruit set ultimately lower yield.


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