A novel source of cytoplasmic male sterility in Calibrachoa pubescens

Colombo, N.; Coviella, A.; Hagiwara, Juan Carlos

doi:10.14295/oh.v23i3.1061

Cited by 4 publications

(3 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Fluorescein diacetate accumulates inside the cytoplasm of viable pollen grains with intact plasma membranes, upon hydrolysis to fluorescein by intracellular esterases through the fluorochromatic reaction (FCR) (Heslop-Harrison and Heslop-Harrison 1970 ), whereas propidium iodide labels pollen wall following its higher affinity for pectins (Rounds et al 2011 ). The same technique was also used for the assessment of pollen sterility levels (Colombo et al 2017 ). In our study, different approaches for the quantitative evaluation of pollen quality using the FDA/PI labelling were tested.…”

Section: Introductionmentioning

confidence: 99%

Quantitative methods in microscopy to assess pollen viability in different plant taxa

et al. 2020

View full text Add to dashboard Cite

High-quality pollen is a prerequisite for plant reproductive success. Pollen viability and sterility can be routinely assessed using common stains and manual microscope examination, but with low overall statistical power. Current automated methods are primarily directed towards the analysis of pollen sterility, and high throughput solutions for both pollen viability and sterility evaluation are needed that will be consistent with emerging biotechnological strategies for crop improvement. Our goal is to refine established labelling procedures for pollen, based on the combination of fluorescein (FDA) and propidium iodide (PI), and to develop automated solutions for accurately assessing pollen grain images and classifying them for quality. We used open-source software programs (CellProfiler, CellProfiler Analyst, Fiji and R) for analysis of images collected from 10 pollen taxa labelled using FDA/PI. After correcting for image background noise, pollen grain images were examined for quality employing thresholding and segmentation. Supervised and unsupervised classification of per-object features was employed for the identification of viable, dead and sterile pollen. The combination of FDA and PI dyes was able to differentiate between viable, dead and sterile pollen in all the analysed taxa. Automated image analysis and classification significantly increased the statistical power of the pollen viability assay, identifying more than 75,000 pollen grains with high accuracy (R2 = 0.99) when compared to classical manual counting. Overall, we provide a comprehensive set of methodologies as baseline for the automated assessment of pollen viability using fluorescence microscopy, which can be combined with manual and mechanized imaging systems in fundamental and applied research on plant biology. We also supply the complete set of pollen images (the FDA/PI pollen dataset) to the scientific community for future research.

show abstract

Section: Introductionmentioning

confidence: 99%

Quantitative methods in microscopy to assess pollen viability in different plant taxa

et al. 2020

View full text Add to dashboard Cite

show abstract

“…Calibrachoa es un género de plantas perteneciente a la familia Solanaceae originario de América del Sur, específicamente de Brasil, Argentina y Uruguay (Tsukamoto et al, 2002), que tiene importancia económica en el mercado mundial de plantas ornamentales por su frecuente uso en canteros y macetas (Colombo et al, 2017) debido a la variedad de colores y a la atractiva forma de sus flores (Milicia et al, 2016).…”

Section: Introductionunclassified

PROTOCOLO DE REGENERACIÓN DE Calibrachoa var. PAMPA SALMÓN INTA A PARTIR DE MERISTEMAS APICALES

Tombion¹,

Kato²,

Soto³

2020

Chil. j. agric. anim. sci.

View full text Add to dashboard Cite

Calibrachoa, perteneciente a la familia Solanaceae, es un género nativo de América del Sur. Las variedades comerciales de esta planta tienen un alto impacto en el mercado de plantas ornamentales por la atractiva forma y color de sus flores, y por su capacidad para adaptarse al uso en canteros y macetas. El objetivo de esta investigación fue desarrollar un protocolo eficiente para la propagación in vitro de Calibrachoa var. Pampa Salmón INTA a partir de meristemas apicales. La combinación de MS ½ + 0,01 mg L-1 BAP resultó en la obtención de un alto porcentaje de regeneración de plantas completas, además de una mayor producción aérea y radical que, consecuentemente, permitieron una alta tasa de sobrevivencia de plantas luego de su transferencia al invernáculo.

show abstract

“…CMS/Rf systems have been widely used in plant breeding to increase the efficiency of hybrid production in several crops (Bohra et al, 2016;Kim and Zhang, 2018). In the case of ornamental species CMS is a desirable trait for breeding purposes to produce hybrid seed, to increase flowering duration, to avoid pollen allergens and to control invasiveness (Colombo et al, 2017).…”

Section: Introductionmentioning

confidence: 99%

PCR identification of Petunia male sterile cytoplasm

Colombo

Hagiwara²

2019

Ornam. Hortic.

Self Cite

View full text Add to dashboard Cite

Petunia is a very important ornamental plant with a broad range of flower colour and size, and most of the cultivars grown are propagated through seeds. Cytoplasmic male sterility (CMS) is a maternally inherited character determined by mitochondrial genes that results in impaired pollen development. The unique and well characterized male sterile cytoplasm in Petunia is a valuable resource for hybrids production because it prevents self-fertilization of mother plants and ensures the purity of F1s. Introgression of the male sterile cytoplasm in elite lines of Petunia is achieved following a backcross scheme and can be assisted using molecular markers associated to the trait of interest. The objective of this study was to develop a molecular marker to identify the male sterile cytoplasm of Petunia. A PCR-based marker amplifying a region of the mitochondrial CMS-associated urfS only in the male sterile plants was designed. Results showed differential PCR amplification of a ≈ 600 bp product in plants carrying male sterile cytoplasm in four Petunia species and their F1s and BC1 generations. A multiplex PCR reaction was subsequently set up, adding specific primers amplifying a ≈ 800 bp product from the conserved region trnT-trn-L of the chloroplast genome as a positive control in order to unambiguously identify the cytoplasm types as normal or sterile. A rapid, simple and precise molecular marker is now available for assisting breeding of F1 hybrids in Petunia.

show abstract

A novel source of cytoplasmic male sterility in Calibrachoa pubescens

Cited by 4 publications

References 14 publications

Quantitative methods in microscopy to assess pollen viability in different plant taxa

Quantitative methods in microscopy to assess pollen viability in different plant taxa

PROTOCOLO DE REGENERACIÓN DE Calibrachoa var. PAMPA SALMÓN INTA A PARTIR DE MERISTEMAS APICALES

PCR identification of Petunia male sterile cytoplasm

Contact Info

Product

Resources

About