Optimizing pollencounter for high throughput phenotyping of pollen quality in tomatoes
The macro “PollenCounter” in ImageJ was initially developed to assess pollen viability in grapevine. We set out to see if PollenCounter could be used to assess pollen number and viability in tomatoes. • We tested different optimization scenarios by adjusting the pollen size (100–900, 200–900 pixel2)...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2020-01-01
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| Series: | MethodsX |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2215016120301977 |
| _version_ | 1818965211028127744 |
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| author | Mathieu Anatole Tele Ayenan Agyemang Danquah Charles Ampomah-Dwamena Peter Hanson Isaac K. Asante Eric Yirenkyi Danquah |
| author_facet | Mathieu Anatole Tele Ayenan Agyemang Danquah Charles Ampomah-Dwamena Peter Hanson Isaac K. Asante Eric Yirenkyi Danquah |
| author_sort | Mathieu Anatole Tele Ayenan |
| collection | DOAJ |
| description | The macro “PollenCounter” in ImageJ was initially developed to assess pollen viability in grapevine. We set out to see if PollenCounter could be used to assess pollen number and viability in tomatoes. • We tested different optimization scenarios by adjusting the pollen size (100–900, 200–900 pixel2) and circularity of pollen grains (0.4–1, 0.5–1, and 0.6–1) on 31 microscopic images of stained tomato pollen. Both total pollen number and proportion of viable pollen were positively and significantly correlated with the outputs from manual counting. The scenario with 100–900 pixel2 pollen size and 0.4–1 circularity had the highest association for pollen number (r = 0.99) and pollen viability (r = 0.86). PollenCounter is 32-fold faster than manual counting. • We added a command to the macro to automatically save the outputs containing the number of total and viable pollen, avoiding transcription errors inherent to manual counting. • We successfully applied the optimized PollenCounter to discriminate tomato genotypes based on pollen number and pollen viability under heat stress. Our results show that PollenCounter, as an open-access macro, can be customized and improved to meet users’ needs. The use of PollenCounter can save time and money in pollen quality assessment. We outline the steps to optimize the macro for other samples or crop species. The optimized macro could allow efficient screening of a large germplasm collection for pollen thermo-tolerance and selection of best thermo-tolerant individuals in breeding programs. |
| first_indexed | 2024-12-20T13:13:24Z |
| format | Article |
| id | doaj.art-38890c65def044e7b2e30f06928cd33d |
| institution | Directory Open Access Journal |
| issn | 2215-0161 |
| language | English |
| last_indexed | 2024-12-20T13:13:24Z |
| publishDate | 2020-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | MethodsX |
| spelling | doaj.art-38890c65def044e7b2e30f06928cd33d2022-12-21T19:39:37ZengElsevierMethodsX2215-01612020-01-017100977Optimizing pollencounter for high throughput phenotyping of pollen quality in tomatoesMathieu Anatole Tele Ayenan0Agyemang Danquah1Charles Ampomah-Dwamena2Peter Hanson3Isaac K. Asante4Eric Yirenkyi Danquah5West Africa Centre for Crop Improvement, College of Basic and Applied Science, University of Ghana, Legon, Ghana; Corresponding author.West Africa Centre for Crop Improvement, College of Basic and Applied Science, University of Ghana, Legon, GhanaThe New Zealand Institute for Plant & Food Research Limited (PFR), Auckland, New ZealandWorld Vegetable Center, West and Central Africa – Coastal and Humid Regions, IITA-Benin Campus, Abomey-Calavi, Republic of BeninUniversity of Ghana, Department of Plant Biology and Environmental Science, Legon, GhanaWest Africa Centre for Crop Improvement, College of Basic and Applied Science, University of Ghana, Legon, GhanaThe macro “PollenCounter” in ImageJ was initially developed to assess pollen viability in grapevine. We set out to see if PollenCounter could be used to assess pollen number and viability in tomatoes. • We tested different optimization scenarios by adjusting the pollen size (100–900, 200–900 pixel2) and circularity of pollen grains (0.4–1, 0.5–1, and 0.6–1) on 31 microscopic images of stained tomato pollen. Both total pollen number and proportion of viable pollen were positively and significantly correlated with the outputs from manual counting. The scenario with 100–900 pixel2 pollen size and 0.4–1 circularity had the highest association for pollen number (r = 0.99) and pollen viability (r = 0.86). PollenCounter is 32-fold faster than manual counting. • We added a command to the macro to automatically save the outputs containing the number of total and viable pollen, avoiding transcription errors inherent to manual counting. • We successfully applied the optimized PollenCounter to discriminate tomato genotypes based on pollen number and pollen viability under heat stress. Our results show that PollenCounter, as an open-access macro, can be customized and improved to meet users’ needs. The use of PollenCounter can save time and money in pollen quality assessment. We outline the steps to optimize the macro for other samples or crop species. The optimized macro could allow efficient screening of a large germplasm collection for pollen thermo-tolerance and selection of best thermo-tolerant individuals in breeding programs.http://www.sciencedirect.com/science/article/pii/S2215016120301977Heat-toleranceImageJSolanum lycopersicumPollen viability |
| spellingShingle | Mathieu Anatole Tele Ayenan Agyemang Danquah Charles Ampomah-Dwamena Peter Hanson Isaac K. Asante Eric Yirenkyi Danquah Optimizing pollencounter for high throughput phenotyping of pollen quality in tomatoes MethodsX Heat-tolerance ImageJ Solanum lycopersicum Pollen viability |
| title | Optimizing pollencounter for high throughput phenotyping of pollen quality in tomatoes |
| title_full | Optimizing pollencounter for high throughput phenotyping of pollen quality in tomatoes |
| title_fullStr | Optimizing pollencounter for high throughput phenotyping of pollen quality in tomatoes |
| title_full_unstemmed | Optimizing pollencounter for high throughput phenotyping of pollen quality in tomatoes |
| title_short | Optimizing pollencounter for high throughput phenotyping of pollen quality in tomatoes |
| title_sort | optimizing pollencounter for high throughput phenotyping of pollen quality in tomatoes |
| topic | Heat-tolerance ImageJ Solanum lycopersicum Pollen viability |
| url | http://www.sciencedirect.com/science/article/pii/S2215016120301977 |
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