Low CO<sub>2</sub> Levels Are Detrimental for In Vitro Plantlets through Disturbance of Photosynthetic Functionality and Accumulation of Reactive Oxygen Species
Photosynthesis of plantlets in tissue culture containers is not considered important, compared to photosynthesis of ex vitro plants, due to the exogenous source of carbohydrates present in tissue culture media. However, CO<sub>2</sub> starvation can generate a burst of reactive oxygen sp...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-01-01
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| Series: | Horticulturae |
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| Online Access: | https://www.mdpi.com/2311-7524/8/1/44 |
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| author | Naser Askari Sasan Aliniaeifard Richard G. F. Visser |
| author_facet | Naser Askari Sasan Aliniaeifard Richard G. F. Visser |
| author_sort | Naser Askari |
| collection | DOAJ |
| description | Photosynthesis of plantlets in tissue culture containers is not considered important, compared to photosynthesis of ex vitro plants, due to the exogenous source of carbohydrates present in tissue culture media. However, CO<sub>2</sub> starvation can generate a burst of reactive oxygen species (ROS). We examined this phenomenon in tissue culture, since CO<sub>2</sub> levels may become very low during the light period. The research was carried out with lily scales, regenerating adventitious bulblets, and with <i>Arabidopsis</i> seedlings. CO<sub>2</sub> starvation was achieved by placing a small vial of concentrated KOH solution in the culture container. CO<sub>2</sub> removal reduced the growth of regenerated lily bulblets by 33% or 23%, with or without sucrose in the medium, respectively. In <i>Arabidopsis</i> seedlings, CO<sub>2</sub> removal decreased growth by 50% or 78% in the presence or absence of sucrose in the medium, respectively. Therefore, the addition of sucrose as a replacement for photosynthesis resulted in only partial recovery of growth. Staining with nitroblue tetrazolium (NBT) showed little to no ROS in ex vitro growing seedlings, while abundant ROS were detected in seedlings grown under in vitro CO<sub>2</sub> starvation. Seedlings grown under normal tissue culture conditions (no CO<sub>2</sub> withdrawal) showed low levels of ROS. In lily tissue culture, CO<sub>2</sub> starvation decreased the maximum quantum efficiency of photosystem II (F<sub>v</sub>/F<sub>m</sub>) from 0.69 to 0.60, and in <i>Arabidopsis</i> from 0.76 to 0.62. F<sub>v</sub>/F<sub>m</sub> of ex vitro lily and <i>Arabidopsis</i> seedlings was 0.77 and 0.79, respectively. This is indicative of a disturbance in photosynthesis functionality and the occurrence of in vitro stress under reduced CO<sub>2</sub> concentrations. We conclude that poor growth, in the absence of CO<sub>2</sub>, was partly due to strongly reduced photosynthesis, while the detrimental effects were most likely due to a burst of ROS. |
| first_indexed | 2024-03-10T01:21:48Z |
| format | Article |
| id | doaj.art-4a643a76833242cbbedc66d2ee017e7b |
| institution | Directory Open Access Journal |
| issn | 2311-7524 |
| language | English |
| last_indexed | 2024-03-10T01:21:48Z |
| publishDate | 2022-01-01 |
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| series | Horticulturae |
| spelling | doaj.art-4a643a76833242cbbedc66d2ee017e7b2023-11-23T13:57:43ZengMDPI AGHorticulturae2311-75242022-01-01814410.3390/horticulturae8010044Low CO<sub>2</sub> Levels Are Detrimental for In Vitro Plantlets through Disturbance of Photosynthetic Functionality and Accumulation of Reactive Oxygen SpeciesNaser Askari0Sasan Aliniaeifard1Richard G. F. Visser2Plant Breeding, Wageningen University & Research, P.O. Box 386, 6700 AJ Wageningen, The NetherlandsPhotosynthesis Laboratory, Department of Horticulture, Aburaihan Campus, University of Tehran, Tehran 33916-53755, IranPlant Breeding, Wageningen University & Research, P.O. Box 386, 6700 AJ Wageningen, The NetherlandsPhotosynthesis of plantlets in tissue culture containers is not considered important, compared to photosynthesis of ex vitro plants, due to the exogenous source of carbohydrates present in tissue culture media. However, CO<sub>2</sub> starvation can generate a burst of reactive oxygen species (ROS). We examined this phenomenon in tissue culture, since CO<sub>2</sub> levels may become very low during the light period. The research was carried out with lily scales, regenerating adventitious bulblets, and with <i>Arabidopsis</i> seedlings. CO<sub>2</sub> starvation was achieved by placing a small vial of concentrated KOH solution in the culture container. CO<sub>2</sub> removal reduced the growth of regenerated lily bulblets by 33% or 23%, with or without sucrose in the medium, respectively. In <i>Arabidopsis</i> seedlings, CO<sub>2</sub> removal decreased growth by 50% or 78% in the presence or absence of sucrose in the medium, respectively. Therefore, the addition of sucrose as a replacement for photosynthesis resulted in only partial recovery of growth. Staining with nitroblue tetrazolium (NBT) showed little to no ROS in ex vitro growing seedlings, while abundant ROS were detected in seedlings grown under in vitro CO<sub>2</sub> starvation. Seedlings grown under normal tissue culture conditions (no CO<sub>2</sub> withdrawal) showed low levels of ROS. In lily tissue culture, CO<sub>2</sub> starvation decreased the maximum quantum efficiency of photosystem II (F<sub>v</sub>/F<sub>m</sub>) from 0.69 to 0.60, and in <i>Arabidopsis</i> from 0.76 to 0.62. F<sub>v</sub>/F<sub>m</sub> of ex vitro lily and <i>Arabidopsis</i> seedlings was 0.77 and 0.79, respectively. This is indicative of a disturbance in photosynthesis functionality and the occurrence of in vitro stress under reduced CO<sub>2</sub> concentrations. We conclude that poor growth, in the absence of CO<sub>2</sub>, was partly due to strongly reduced photosynthesis, while the detrimental effects were most likely due to a burst of ROS.https://www.mdpi.com/2311-7524/8/1/44lilyarabidopsistissue cultureCO<sub>2</sub> starvationROS |
| spellingShingle | Naser Askari Sasan Aliniaeifard Richard G. F. Visser Low CO<sub>2</sub> Levels Are Detrimental for In Vitro Plantlets through Disturbance of Photosynthetic Functionality and Accumulation of Reactive Oxygen Species Horticulturae lily arabidopsis tissue culture CO<sub>2</sub> starvation ROS |
| title | Low CO<sub>2</sub> Levels Are Detrimental for In Vitro Plantlets through Disturbance of Photosynthetic Functionality and Accumulation of Reactive Oxygen Species |
| title_full | Low CO<sub>2</sub> Levels Are Detrimental for In Vitro Plantlets through Disturbance of Photosynthetic Functionality and Accumulation of Reactive Oxygen Species |
| title_fullStr | Low CO<sub>2</sub> Levels Are Detrimental for In Vitro Plantlets through Disturbance of Photosynthetic Functionality and Accumulation of Reactive Oxygen Species |
| title_full_unstemmed | Low CO<sub>2</sub> Levels Are Detrimental for In Vitro Plantlets through Disturbance of Photosynthetic Functionality and Accumulation of Reactive Oxygen Species |
| title_short | Low CO<sub>2</sub> Levels Are Detrimental for In Vitro Plantlets through Disturbance of Photosynthetic Functionality and Accumulation of Reactive Oxygen Species |
| title_sort | low co sub 2 sub levels are detrimental for in vitro plantlets through disturbance of photosynthetic functionality and accumulation of reactive oxygen species |
| topic | lily arabidopsis tissue culture CO<sub>2</sub> starvation ROS |
| url | https://www.mdpi.com/2311-7524/8/1/44 |
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