Comparison of Two Staining Methods for Pollen Viability Studies in Sugarcane

Melloni, Maria Letícia Guindalini; Scarpari, Maximiliano Salles; Mendonça, Jeremias R. de; Pérecin, Dilermando; Landell, Marcos Guimarães de Andrade; Pinto, Luciana Rossini

doi:10.1007/s12355-012-0185-6

Cited by 20 publications

(9 citation statements)

References 12 publications

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“…The iodine-staining method relies on pollen reserves and does not guarantee effective germination; therefore, the iodine-staining method might overestimate pollen viability (Rodriguez-Riano and Dafni, 2000). In addition, the correlation between pollen viability assessed through staining methods and germination on culture medium was low when it was evaluated in different sugarcane cultivars (Melloni et al, 2013). Therefore, the pollen viability scores that were assigned here to the inflorescences that were involved in the polycross may not be correct, especially the scores of 3 and 4 that were assigned to cultivars IAC91-1099 and RB86-7515, respectively, both of which were considered to be a pollen donor (male).…”

Section: Discussionmentioning

confidence: 99%

“…The knowledge of the selfing rate, whether from biparental or polycross families, is critical in breeding programs to correctly evaluate the fam-ily's performance and thus determine the correct breeding value of the parents involved in the respective cross (McIntyre and Jackson, 2001). All of these studies were conducted with favorable environmental conditions of sugarcane crossing, i.e., high air humidity (above 85%) and temperature ranging from 18 to 31°C with an optimal temperature of 27°C (Melloni et al, 2013), which naturally occurs at the northeast region of Brazil, where the sugarcane breeding cross stations are located.…”

Section: Introductionmentioning

confidence: 99%

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Selfing rate estimation in sugarcane under unfavorable natural conditions of crossing by using microsatellite markers

Melloni¹,

Scarpari²,

Pinto³

et al. 2014

Genet. Mol. Res.

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ABSTRACT. The self-fertilization or selfing rate estimation using microsatellite markers and its impact on survival and selection rate were evaluated in families derived from polycrosses that involved parents that were widely used in sugarcane breeding in Brazil. These factors were evaluated under unfavorable natural conditions of flowering and crossing. After the germination test, the viable progeny were taken to the field for survival rate evaluation (4, 6, and 10 months) and phenotypic selection at plant cane. The selfing rate estimate based on microsatellite markers present in the progeny and absent in their female parent was 98.5 and 0% for the polycross families derived from IACSP95-5000 and SP89-1115, respectively. The survival and selection rates in the last 2 evaluations were higher for the SP89-1115 outcrossed family than the IACSP95-5000 selfed family. The IACSP95-5000 cultivar excelled either as pollen donor with fertilization capability or viable seed production even under unfavorable natural conditions of crossing. The Selfing rate in sugarcane environment influence (temperature and humidity) had an important role during the polycross.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Selfing rate estimation in sugarcane under unfavorable natural conditions of crossing by using microsatellite markers

Melloni¹,

Scarpari²,

Pinto³

et al. 2014

Genet. Mol. Res.

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“…Different techniques are cited in the literature to investigate the occurrence of viable pollens in several species and these are grouped in direct methods, such as in vitro germination [7,8,9] and in vivo [9,10,11] or indirect based in cytological parameters such as staining [9,11,12,13].…”

Section: Introductionmentioning

confidence: 99%

Pollen Viability in Eggplant Using Colorimetric and In vitro Techniques

Valadares

Lima

Nóbrega

et al. 2019

JEAI

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The causes of fruit abortion, in eggplants, may be related to the absence of viable pollen under high temperatures, common in the Northeast region of Brazil, resulting in a reduction in the number of fruits per plant and consequently in productivity. The objective of this study was to evaluate the techniques of identification of PV – pollen viability in eggplant, as well as to correlate these results with the FFI – Fruit Fixation Index, NFP – Number of Fruits per Plant and PP – Production per Plant obtained in cultivation under high temperatures. The experiment was conducted at the Federal Rural University of Pernambuco, Recife, Brazil, between September and December 2017. The experiment design was a randomized block design with four replications, in the 7 x 4 factorial scheme (7 genotypes x 4 evaluation of PV) containing four plants per experimental plot. The results showed that the different techniques used detected viable pollens in the genotypes evaluated in smaller and in larger proportions. The highest percentages were observed in the treatments with CA – Carmine Acetic and AS – Alexander Solution and these overestimated the PV (>90%). In the test with TTC – 2,3,5-triphenyltetrazolium chloride (25%), the viability estimation was equivalent to the in vitro germination test, however the genetic correlations were null and/or very low between these techniques and the FFI, NFP and PP traits. The genetic correlations between the results obtained with CA and AS with FFI, NFP and PP under conditions of high temperatures were high and positive and suggest that the selection based on PV only, except with the use of TTC solution, may be efficient for indirect selection of genotypes with high FFI, NFP and PP.

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“…In Manihot, the most commonly employed method for establishing pollen viability is a colorimetric assay using different types of stains, such as acetic carmine, Lugol's solution, Alexander's solution and acetic orcein (Silva et al, 2001;. However, no correspondence has been observed between the colorimetric tests and the in vivo viability tests, reinforcing theories suggested by some authors regarding the inefficiency of these stains as proof of pollen viability (Rodriguez-Riano and Dafni 2000; Melloni et al, 2013). Aniline blue is a stain specific for callose located on the inner layer of the pollen tube, which fluoresces under ultraviolet light.…”

Section: Introductionmentioning

confidence: 95%

Use of aniline blue stain to observing pollen tubes development in different Manihot Mill. species

Vieira¹,

Santana²,

Alves³

et al. 2015

Afr. J. Agric. Res.

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The lack of a reliable test to evaluate pollen viability in species of the genus Manihot is one of the primary limitations for cassava genetic improvement. Thus, the objective of the present study was to apply aniline blue stain to observing pollen tubes development in different Manihot Mill. species, in order to estimate the crossability, pollen germination time, pollen tube size and pollen longevity. Pollen grains from 2 accessions of the cultivated species M. esculenta spp. esculenta and 4 accessions of 3 wild species (M. esculenta ssp. flabellifolia (Pohl) Cif, M. anomala Pohl, and M. irwinii D. J. Roger and Appan) were used. To visualize the pollen tubes, pollinated stigmas was deposited on a glass slide with 3 drops of aniline blue stain and transferred to an optical fluorescence microscope, where they were visualized and photographed using the 10x ocular lens. The pollen crossability rate was estimated by the percentage of germinated pollen, with well-developed pollen tubes in relation to the total number of pollen grains counted per stigma. The experimental design was entirely random with 3 repetitions, each repetition consisting of 100 pollen grains. In addition to the estimation of crossability, the germination time after pollination, pollen tube size, and pollen longevity were also determined to the accession FLA 029V-01. Significant difference was observed between the crossings. Pollen tubes developed after 45 min of contact between the pollen and the stigma. The average pollen tube size 24 h after pollination was 2.9 mm. The studies examining pollen longevity showed that germination did not occur when the pollen grains were evaluated 24 h after anthesis. Thus, the use of aniline blue stain can be considered an effective method for observing pollen tubes development and studying the pollen morphology, longevity, viability and Manihot crossability.

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Comparison of Two Staining Methods for Pollen Viability Studies in Sugarcane

Cited by 20 publications

References 12 publications

Selfing rate estimation in sugarcane under unfavorable natural conditions of crossing by using microsatellite markers

Selfing rate estimation in sugarcane under unfavorable natural conditions of crossing by using microsatellite markers

Pollen Viability in Eggplant Using Colorimetric and In vitro Techniques

Use of aniline blue stain to observing pollen tubes development in different Manihot Mill. species

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