scholarly journals Technique for In Vitro Pollen Germination and Short-term Pollen Storage in Caladium

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).

1990 ◽  
Vol 115 (3) ◽  
pp. 359-363 ◽  
Author(s):  
I.E. Yates ◽  
Darrell Sparks

Stored pollen from pecan [Carya illinoensis (Wangenh.) C. Koch] was analyzed for in vitro germination, fertilization efficiency, final fruit set, and characteristics of mature fruits. We demonstrate pecan pollen can be stored for several years and set fruit. Pollen stored for 1, 2, and 3 years at -80C and 1 year at -196C retained the capacity for fertilization. Pollen stored at -196C was more viable than pollen stored at -80C, with no significant correlation between length of storage at -80C, as judged by fruit abortions during the second drop. Final fruit set was not affected by pollen storage conditions, except for pollen collected in a season of drought. Fruit set is a more reliable indicator of pollen viability than in vitro germination. With two minor exceptions, fruits produced with stored pollen were similar to those developing after pollination with fresh 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.


HortScience ◽  
2000 ◽  
Vol 35 (6) ◽  
pp. 1151-1152 ◽  
Author(s):  
P. Martínez-Gómez ◽  
T.M. Gradziel ◽  
E. Ortega ◽  
F. Dicenta

Almond [Prunus dulcis (Mill.) D.A. Webb] breeding programs require successful techniques for pollen storage. We studied the pollen viability of two almond cultivars, `Ramillete' and `Desmayo Largueta', during 8 weeks of storage, in conditions that simulated standard situations including storage at 4, 22, and 4 °C alternating with 22 °C (4 °C/22 °C). Viability remained at 60% or more for 2 weeks under all three conditions. After the second week, germination capacity decreased rapidly at 22 °C, but remained above 50% for as long as 8 weeks at 4 °C or 4 °C/22 °C.


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.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Andrej Kormuťák ◽  
Peter Bolecek ◽  
Martin Galgóci ◽  
Dušan Gömöry

AbstractPollen storage belongs among the most important activities associated with pollen handling. It overcomes the differences in pollen shedding and ovule receptivity during controlled pollination experiments. It is especially important for species like common juniper (Juniperus communis L.) with an extremely low quality of seeds due to pollination failure. Additionally, it is a substantial part of germplasm preservation programmes in pollen banks. In the present paper, the effect of short-term storage of pollen was studied using pollen samples from five shrubs in an in vitro germination test. Two temperature regimes were tested. The pollen viability of freshly collected pollen varied considerably between individual shrubs, exhibiting 67.3–88.6% germination rate and 248.0–367.3 µm of pollen tubes. Storage at + 4 °C for four months was accompanied by a profound decline in pollen viability. The germination percentage was reduced to 49.2–75.2% and the pollen tube length to 32.5–69.0%, depending on individual shrubs. The corresponding decline in pollen viability characteristics during storage at − 20 °C was only negligible in two of the tested shrubs. In the remaining three shrub samples, an increase in germination percentage was observed. Pollen tube growth responded more sensitively to freezing, but, on average, the decrease in length was lower than that at + 4 °C. The rate of reduction in pollen tube length varied between 11.5 and 45.4%. Cytological events accompanying in vitro germination of freezer-stored pollen exhibited some delay in releasing the exine from pollen grains during the early stages of germination as compared with freshly collected pollen. In conclusion, short-term storage of the common juniper pollen in a freezer is better for the preservation of its viability than storage at + 4 °C.


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.


HortScience ◽  
2006 ◽  
Vol 41 (4) ◽  
pp. 1020C-1020
Author(s):  
Ann M. Chanon ◽  
Pablo S. Jourdan ◽  
Joseph C. Scheerens

The genus Aesculus (buckeyes and/or horsechestnuts) is composed of 13 species and a number of interspecific hybrids. Pollen from 11 genotypes from five Aesculus species and the hybrid Aesculus ×carnea were used to develop an in-vitro germination test to evaluate pollen viability under various storage treatments. This test was optimized using samples of both fresh pollen and pollen that had been stored up to 1 year. The most effective medium contained 20% sucrose, 100 mg·L-1 H2BO3, 150 mg·L-1 Ca(NO3)2, and 1% agar. The highest germination percentage was observed at 15 °C across all storage treatments. Fresh pollen germinated in excess of 80% over a wide range of germination temperatures. Based on this, all specimens studied would be good pollen parents. The differences in pollen germination between storage at -20 and -80 °C were nonsignificant, but the duration of the storage period was highly significant. At 3 months, viability remained above 60% for four of the six species/hybrid tested. However, at 12 months, all pollen tested dropped below the threshold for good fruit set based on in-vitro pollen germination. Based on these observations, short-term pollen storage may permit crosses between parents with temporally separate flowering phenologies. However, conventional storage procedures are inadequate to maintain pollen collected from a male parent for crosses in subsequent growing seasons.


2000 ◽  
Vol 125 (2) ◽  
pp. 265-270 ◽  
Author(s):  
A.M.S. Nyomora ◽  
P.H. Brown ◽  
K. Pinney ◽  
V.S. Polito

The effect of boron (B) on in vivo and in vitro development of almond [Prunus dulcis (Mill.) D.A. Webb (syn. P. amygdalus Batsch)] pollen and pollen tubes and the resultant effect on fruit set was studied in mature trees. The cultivars Mono (pistil donor) and Butte (pollinizer) in an orchard with low soil B in Fresno, California were sprayed with B at 0, 0.8, 1.7, or 2.5 kg·ha-1 during Fall 1993. Pollen viability as indicated by the fluorescein diacetate method (FDA) was >85% and was not affected by field-applied B, however, in vivo pollen germination and tube growth were enhanced by foliar-applied B. More effect of applied B on in vivo growth appeared as pollen tubes progressed toward the ovary. For in vitro germination, foliar-applied B reduced bursting of tubes, and addition of B to the culture media significantly increased pollen germination and pollen tube growth.


2012 ◽  
Vol 39 (12) ◽  
pp. 1009 ◽  
Author(s):  
Viola Devasirvatham ◽  
Pooran M. Gaur ◽  
Nalini Mallikarjuna ◽  
Raju N. Tokachichu ◽  
Richard M. Trethowan ◽  
...  

High temperature during the reproductive stage in chickpea (Cicer arietinum L.) is a major cause of yield loss. The objective of this research was to determine whether that variation can be explained by differences in anther and pollen development under heat stress: the effect of high temperature during the pre- and post-anthesis periods on pollen viability, pollen germination in a medium, pollen germination on the stigma, pollen tube growth and pod set in a heat-tolerant (ICCV 92944) and a heat-sensitive (ICC 5912) genotype was studied. The plants were evaluated under heat stress and non-heat stress conditions in controlled environments. High temperature stress (29/16°C to 40/25°C) was gradually applied at flowering to study pollen viability and stigma receptivity including flower production, pod set and seed number. This was compared with a non-stress treatment (27/16°C). The high temperatures reduced pod set by reducing pollen viability and pollen production per flower. The ICCV 92944 pollen was viable at 35/20°C (41% fertile) and at 40/25°C (13% fertile), whereas ICC 5912 pollen was completely sterile at 35/20°C with no in vitro germination and no germination on the stigma. However, the stigma of ICC 5912 remained receptive at 35/20°C and non-stressed pollen (27/16°C) germinated on it during reciprocal crossing. These data indicate that pollen grains were more sensitive to high temperature than the stigma in chickpea. High temperature also reduced pollen production per flower, % pollen germination, pod set and seed number.


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