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Proline synthesis in developing microspores is required for pollen development and fertility
Roberto Mattioli1, Marco Biancucci1,2, Amira El Shall1
1Department of Biology and Biotechnology, Sapienza University of Rome, P.le A. Moro 5, 00185, Rome, Italy.
BMC Plant Biology
|December 19, 2018
Summary
Plant pollen development requires proline. Local proline synthesis, not transport, is crucial for fertile pollen, especially in later stages of microspore development and mature pollen grains.
Area of Science:
- Plant reproductive biology
- Molecular genetics
- Amino acid metabolism
Background:
- Proline accumulation in pollen is vital for plant fertility.
- Disruption of proline synthesis leads to microspore abortion in Arabidopsis.
- The role of local proline biosynthesis versus transport in pollen development was previously unclear.
Purpose of the Study:
- To investigate whether local proline biosynthesis or transport is essential for fertile pollen development in Arabidopsis.
- To identify the specific site of proline synthesis required for successful microspore development.
Main Methods:
- Analyzed expression patterns of proline biosynthetic genes PYRROLINE-5-CARBOXYLATE SYNTHETASE 1 & 2 (P5CS1 & 2) in Arabidopsis anthers.
- Generated transgenic Arabidopsis lines expressing P5CS2 under different promoters (CaMV 35S, tapetum-specific Ltp12, pollen-specific At5g17340) in a p5cs1/p5cs1 p5cs2/P5CS2 mutant background.
- Confirmed promoter specificity using β-glucuronidase (GUS) assays and measured proline levels in pollen and anthers.
Main Results:
- Both P5CS1 and P5CS2 showed strong expression in developing microspores and pollen grains.
- Pollen-specific expression of P5CS2 fully restored proline content, morphology, and fertility in mutant pollen.
- Expression under CaMV 35S or tapetum-specific promoters resulted in only partial restoration of pollen development and fertility.
Conclusions:
- Proline transport cannot meet the demands of the male germline cells.
- Local proline biosynthesis within the microspores/pollen grains is essential for their development and fertility.
- Successful pollen development relies on proline synthesis during late microspore stages and in mature pollen.
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