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In vitro microspore selection in maize anther culture with oxidative-stress stimulators
1Agricultural Research Institute, Hungarian Academy of Sciences, Martonvásár, Hungary.
Abstract:
In order to produce doubled-haploid maize plants tolerant of oxidative stress, in vitro microspore selection was carried out in anther culture with reactive oxygen species (ROS) progenitors such as paraquat, menadione, tert-butylhydroperoxide (t-BHP), and methionine combined with riboflavin. All the ROS progenitors reduced the anther induction, the formation of microspore-derived structures, and their regeneration potential. Abnormal cell divisions and progeny cell degradation could be observed during the development of microspores treated with ROS progenitors. Menadione and t-BHP influenced the microspore developmental pathway, as menadione induced the formation of embryoids, while t-BHP increased the proportion of calli in the microspore-derived structures. As the result of in vitro selection, 15, 10, 10, and 3 fertile doubled-haploid plants were obtained in cultures treated with paraquat, t-BHP, methionine combined with riboflavin, and menadione, respectively.
Insights
This study developed oxidative stress-tolerant doubled-haploid maize plants using in vitro microspore selection with reactive oxygen species (ROS) progenitors. ROS treatments reduced regeneration but yielded stress-tolerant plants, demonstrating a novel selection strategy.
Area of Science:
- Plant Biotechnology
- Crop Science
- Genetics
Background:
- Doubled-haploid plants are crucial for accelerated crop breeding.
- Developing oxidative stress tolerance in crops is essential for climate resilience.
- In vitro selection methods can enhance desirable traits in plants.
Purpose of the Study:
- To produce doubled-haploid maize plants with enhanced tolerance to oxidative stress.
- To investigate the effects of reactive oxygen species (ROS) progenitors on microspore development and regeneration.
- To establish an efficient in vitro selection protocol for stress-tolerant maize.
Main Methods:
- Anther culture of maize was employed for microspore development.
- Reactive oxygen species (ROS) progenitors (paraquat, menadione, t-BHP, methionine/riboflavin) were used for in vitro selection.
- Microspore-derived structures were analyzed for developmental abnormalities and regeneration potential.
Main Results:
- All ROS progenitors negatively impacted anther induction and regeneration.
- Abnormal cell divisions and degradation were observed in ROS-treated microspores.
- Specific ROS progenitors differentially affected developmental pathways: menadione induced embryoids, t-BHP increased calli.
- Fertile doubled-haploid plants tolerant to oxidative stress were successfully obtained.
Conclusions:
- In vitro selection using ROS progenitors is a viable strategy for generating oxidative stress-tolerant doubled-haploid maize.
- ROS treatments can modulate microspore developmental pathways, influencing the type of regenerated structures.
- This approach offers a promising avenue for breeding climate-resilient maize varieties.
