Staining and fluorescent microscopy methods for pollen viability determination in sunflower and other plant species

Atlagić, Jovanka; Terzić, S.; Marjanović-Jeromela, Ana

doi:10.1016/j.indcrop.2011.06.012

Cited by 30 publications

(19 citation statements)

References 9 publications

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“…Prior to both preparation methods the flower heads were laid into ethanol 70 % (prepared from ethanol 96 % vol, VWR, Fontenay-sous-Bois, France) for about 24 h. Both methods were adapted following basically described procedures in Kearns and Inouye (1993). Very similar methods of monitoring pollen germination and pollen tube growth in sun flower by fluorescence microscopy were also described in Atlagic et al (2012).…”

Section: Methodsmentioning

confidence: 99%

Pollen tube growth and self incompatibility in Matricaria recutita

et al. 2015

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Understanding of fertilization procedures and crossing barriers is essential for various plant breeding methods. Growth or inhibition of pollen tubes, therefore, is a crucial issue in terms of propagation, in particular concerning the determination of self incompatible (SI-) plants. In German Chamomile, vegetatively propagated SI-plants would be a highly appreciated breeding tool for the formation of maternal lines for specific crossings. Following the idea to enhance knowledge of aspects of pollination and to develop a fast procedure of determination of SI-plants, two different methods of microscopic analysis of pollen tube growth in pistils were developed. Under light and under fluorescence microscopy diverse stages of pollen tube growth could be made visible. These included growth starts at the stigma, active pollen tubes with regular callose deposits on their way through the style and globular stage embryos in the ovary. Furthermore, SI-tests and selections within six accessions of German Chamomile and among 220 plants were carried out. In this case, self incompatibility (SI) was determined as staying seedless after multiple hand pollination under crispac bag isolation, observed at not less than three flower heads per plant. Diploid varieties showed a tendency to establish SI, rather than their tetraploid relatives, although a high variability between accessions was given. An overall accordance between SI, examined via seed set, and evidence of pollen tubes could not be assessed. Nevertheless, new and detailed insight in fertilization procedures in German chamomile could be gained.

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Section: Methodsmentioning

confidence: 99%

Pollen tube growth and self incompatibility in Matricaria recutita

et al. 2015

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“…Owing to the need to store pollen for crosses where parental lines did not flower simultaneously or interspecific crosses are made, staining methods for pollen viability determination have been developed (Alexander, 1969) and improved for use on sunflower (Atlagić et al, 2012).…”

Section: Pollen Production and Transfermentioning

confidence: 99%

Review of 40 years of research carried out in Serbia on sunflower pollination

Terzić

Miklič

Čanak

2017

OCL

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-Sunflower pollination has been a subject of research for more than 40 years at the Institute of Field and Vegetable Crops in Novi Sad, Serbia. Since the first experiments in 1970s, the importance of climatic conditions was acknowledged and analyzed as a key factor for the performance of plants and pollinators. Climatic conditions, disease and pest problems changed significantly during this period, but sunflower production technology and breeding also advanced making seed production more stable. Nonetheless, the quality of agronomic practices still varies which also reflects on the plant and pollinator performance. Sunflower breeding led to improved germplasm which still maintains high degree of variability for nectar content and pollinator attractivity. Pollinators are essential in crosses involving cytoplasmic male sterile lines, but are also beneficial in hybrid sunflower seed production. Pollinator attractivity is thus an important trait for both breeders and producers and worth to be considered as a quality trait in sunflower breeding programs.Keywords: sunflower / pollination / honey bee / Serbia Résumé -Retour sur 40 années de recherche menées en Serbie sur la pollinisation du tournesol. La pollinisation du tournesol est l'objet de travaux de recherche depuis plus de 40 ans à l'Institut des cultures de plein champ et des cultures légumières de Novi Sad, en Serbie. Depuis les premières expériences conduites dans les années 1970, l'importance des conditions climatiques a été reconnue et analysée comme un facteur clé de la performance des plantes et des insectes pollinisateurs. Les conditions climatiques, les problèmes de maladies et de ravageurs ont considérablement changé au cours de cette période, mais les techniques de production et la sélection du tournesol ont également progressé, ce qui a permis de stabiliser la production de semences. Néanmoins, la qualité des pratiques agronomiques reste encore fluctuante, influant également sur la performance des plantes et celle des insectes pollinisateurs. La sélection du tournesol a conduit à l'amélioration du matériel génétique, qui possède encore un haut degré de variabilité vis-à-vis de la production de nectar et l'attractivité des insectes pollinisateurs. Les insectes pollinisateurs s'avèrent essentiels dans les croisements impliquant des lignées à stérilité mâle cytoplasmique, mais également bénéfiques dans la production de semences hybrides de tournesol. L'attractivité des insectes pollinisateurs représente un caractère important à la fois pour les sélectionneurs et les producteurs et devrait être considérée comme un critère de sélection dans les programmes d'amélioration génétique.Mots clés : tournesol / pollinisation / abeille domestique / Serbie

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“…Results of pollen germination using growth medium are considered more accurate, but still fail to account for pollenpistil interactions. Using fluorescent microscopy to monitor pollen germination and growth of the pollen tube most closely simulates in vivo conditions (Kho and Baër, 1968;Atlagić et al, 2012). Further, visualization of pollen germination and pollen tube growth provides a measure of "cross" incompatibility among species or cultivars.…”

Section: Introductionmentioning

confidence: 99%

“…Jones et al (2007) reported from 0 to 20 seeds/ capsule while other researchers have reported as little as 0-5 seeds/ capsule resulting from crosses among H. macrophylla cultivars (Venturieri et al, 2017). Visualization of pollen germination and pollen tube growth allows for the differentiation between pre and post zygotic reproductive barriers which in turn influences the choice of plant breeding techniques (somatic hybridization or embryo culture) that may be used to mitigate incompatibility (Atlagić et al, 2012).…”

Section: Introductionmentioning

confidence: 99%

Ploidy Level Influences Pollen Tube Growth and Seed Viability in Interploidy Crosses of Hydrangea macrophylla

Alexander

2020

Front. Plant Sci.

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All Hydrangea macrophylla cultivars tested to date are diploid or triploid and triploid H. macrophylla have thicker stems, larger flowers, and larger stoma compared to related diploids. It is unknown whether interploidy crosses between diploid and triploid hydrangeas can be used to develop triploid varieties. The objective of this study was to compare pollen tube development, fruit formation, and seed viability among intra-and interploidy pollinations of H. macrophylla and evaluate the genome size and pollen viability of resultant progeny. By 24 h post-pollination, pollen tubes had reached the ovaries of diploid flowers in 48.7% of samples while pollen tubes reached the ovaries in only 8.7% of triploid flowers (c 2 = 30.6, p < 0.001). By 48 h post-pollination pollen tubes reached the ovaries of diploid and triploid flowers in 72.5% and 53.8% of samples, respectively (c 2 = 26.5, p = 0.001). There was no difference in percentage of flowers with pollen tubes reaching the ovaries in diploid and triploid flowers at 72 h after pollination (c 2 = 7.5, p = 0.60). Analysis of covariance showed that pollen tube length at 24 and 48 h postpollination was significantly influenced by ploidy and flower length of the female parent. Progeny of interploidy crosses was diploid and aneuploid; no triploid progeny were recovered from crosses using triploid parents. Mean genome sizes of offspring from each cross type ranged from 4.56 pg for 2x × 2x offspring to 5.17 pg for 3x × 3x offspring. Estimated ploidy of offspring ranged from 2x for 2x × 2x crosses to 2.4x for 3x × 3x crosses. Pollen stainability rates of flowering offspring using a modified Alexander's stain ranged from 69.6% to 76.4%.

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Staining and fluorescent microscopy methods for pollen viability determination in sunflower and other plant species

Cited by 30 publications

References 9 publications

Pollen tube growth and self incompatibility in Matricaria recutita

Pollen tube growth and self incompatibility in Matricaria recutita

Review of 40 years of research carried out in Serbia on sunflower pollination

Ploidy Level Influences Pollen Tube Growth and Seed Viability in Interploidy Crosses of Hydrangea macrophylla

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