Development of strategies for genetic manipulation and fine‐tuning of a chloroplast retrograde signal 3′‐phosphoadenosine 5′‐phosphate
Abstract Homeostasis of metabolism and regulation of stress‐signaling pathways are important for plant growth. The metabolite 3′‐phosphoadenosine‐5′‐phosphate (PAP) plays dual roles as a chloroplast retrograde signal during drought and high light stress, as well as a toxic by‐product of secondary su...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Wiley
2018-01-01
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| Series: | Plant Direct |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/pld3.31 |
| _version_ | 1819275622529105920 |
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| author | Su Yin Phua Wannarat Pornsiriwong Kai Xun Chan Gonzalo M. Estavillo Barry J. Pogson |
| author_facet | Su Yin Phua Wannarat Pornsiriwong Kai Xun Chan Gonzalo M. Estavillo Barry J. Pogson |
| author_sort | Su Yin Phua |
| collection | DOAJ |
| description | Abstract Homeostasis of metabolism and regulation of stress‐signaling pathways are important for plant growth. The metabolite 3′‐phosphoadenosine‐5′‐phosphate (PAP) plays dual roles as a chloroplast retrograde signal during drought and high light stress, as well as a toxic by‐product of secondary sulfur metabolism, and thus, its levels are regulated by the chloroplastic phosphatase, SAL1. Constitutive PAP accumulation in sal1 mutants improves drought tolerance but can impair growth and alter rosette morphology. Therefore, it is of interest to derive strategies to enable controlled and targeted PAP manipulation that could enhance drought tolerance while minimizing the negative effects on plant growth. We systematically tested the potential and efficiency of multiple established transgenic manipulation tools in altering PAP levels in Arabidopsis. Dexamethasone (dex)‐inducible silencing of SAL1 via hpRNAi [pOpOff:SAL1hpRNAi] yielded reduction in SAL1 transcript and protein levels, yet failed to significantly induce PAP accumulation. Surprisingly, this was not due to insufficient silencing of the inducible system, as constitutive silencing using a strong promoter to drive hpRNAi and amiRNA targeting the SAL1 transcript also failed to increase PAP content or induce a sal1‐like plant morphology despite significantly reducing the SAL1 transcript levels. In contrast, using dex‐inducible expression of SAL1 cDNA to complement an Arabidopsis sal1 mutant successfully modulated PAP levels and restored rosette growth in a dosage‐dependent manner. Results from this inducible complementation system indicate that plants with intermediate PAP levels could have improved rosette growth without compromising its drought tolerance. Additionally, preliminary evidence suggests that SAL1 cDNA driven by promoters of genes expressed specifically during early developmental stages such as ABA‐Insensitive 3 (ABI3) could be another potential strategy for studying and optimizing PAP levels and drought tolerance while alleviating the negative impact of PAP on plant growth in sal1. Thus, we have identified ways that can allow future dissection into multiple aspects of stress and developmental regulation mediated by this chloroplast signal. |
| first_indexed | 2024-12-23T23:27:15Z |
| format | Article |
| id | doaj.art-e24d8837e0154f6c8327ebebc395a193 |
| institution | Directory Open Access Journal |
| issn | 2475-4455 |
| language | English |
| last_indexed | 2024-12-23T23:27:15Z |
| publishDate | 2018-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Plant Direct |
| spelling | doaj.art-e24d8837e0154f6c8327ebebc395a1932022-12-21T17:26:11ZengWileyPlant Direct2475-44552018-01-0121n/an/a10.1002/pld3.31Development of strategies for genetic manipulation and fine‐tuning of a chloroplast retrograde signal 3′‐phosphoadenosine 5′‐phosphateSu Yin Phua0Wannarat Pornsiriwong1Kai Xun Chan2Gonzalo M. Estavillo3Barry J. Pogson4ARC Centre of Excellence in Plant Energy Biology Research School of Biology The Australian National University Canberra ACT AustraliaARC Centre of Excellence in Plant Energy Biology Research School of Biology The Australian National University Canberra ACT AustraliaARC Centre of Excellence in Plant Energy Biology Research School of Biology The Australian National University Canberra ACT AustraliaARC Centre of Excellence in Plant Energy Biology Research School of Biology The Australian National University Canberra ACT AustraliaARC Centre of Excellence in Plant Energy Biology Research School of Biology The Australian National University Canberra ACT AustraliaAbstract Homeostasis of metabolism and regulation of stress‐signaling pathways are important for plant growth. The metabolite 3′‐phosphoadenosine‐5′‐phosphate (PAP) plays dual roles as a chloroplast retrograde signal during drought and high light stress, as well as a toxic by‐product of secondary sulfur metabolism, and thus, its levels are regulated by the chloroplastic phosphatase, SAL1. Constitutive PAP accumulation in sal1 mutants improves drought tolerance but can impair growth and alter rosette morphology. Therefore, it is of interest to derive strategies to enable controlled and targeted PAP manipulation that could enhance drought tolerance while minimizing the negative effects on plant growth. We systematically tested the potential and efficiency of multiple established transgenic manipulation tools in altering PAP levels in Arabidopsis. Dexamethasone (dex)‐inducible silencing of SAL1 via hpRNAi [pOpOff:SAL1hpRNAi] yielded reduction in SAL1 transcript and protein levels, yet failed to significantly induce PAP accumulation. Surprisingly, this was not due to insufficient silencing of the inducible system, as constitutive silencing using a strong promoter to drive hpRNAi and amiRNA targeting the SAL1 transcript also failed to increase PAP content or induce a sal1‐like plant morphology despite significantly reducing the SAL1 transcript levels. In contrast, using dex‐inducible expression of SAL1 cDNA to complement an Arabidopsis sal1 mutant successfully modulated PAP levels and restored rosette growth in a dosage‐dependent manner. Results from this inducible complementation system indicate that plants with intermediate PAP levels could have improved rosette growth without compromising its drought tolerance. Additionally, preliminary evidence suggests that SAL1 cDNA driven by promoters of genes expressed specifically during early developmental stages such as ABA‐Insensitive 3 (ABI3) could be another potential strategy for studying and optimizing PAP levels and drought tolerance while alleviating the negative impact of PAP on plant growth in sal1. Thus, we have identified ways that can allow future dissection into multiple aspects of stress and developmental regulation mediated by this chloroplast signal.https://doi.org/10.1002/pld3.313′‐phosphoadenosine‐5′‐phosphatedroughtgene silencingretrograde signalingRNAiSAL1 |
| spellingShingle | Su Yin Phua Wannarat Pornsiriwong Kai Xun Chan Gonzalo M. Estavillo Barry J. Pogson Development of strategies for genetic manipulation and fine‐tuning of a chloroplast retrograde signal 3′‐phosphoadenosine 5′‐phosphate Plant Direct 3′‐phosphoadenosine‐5′‐phosphate drought gene silencing retrograde signaling RNAi SAL1 |
| title | Development of strategies for genetic manipulation and fine‐tuning of a chloroplast retrograde signal 3′‐phosphoadenosine 5′‐phosphate |
| title_full | Development of strategies for genetic manipulation and fine‐tuning of a chloroplast retrograde signal 3′‐phosphoadenosine 5′‐phosphate |
| title_fullStr | Development of strategies for genetic manipulation and fine‐tuning of a chloroplast retrograde signal 3′‐phosphoadenosine 5′‐phosphate |
| title_full_unstemmed | Development of strategies for genetic manipulation and fine‐tuning of a chloroplast retrograde signal 3′‐phosphoadenosine 5′‐phosphate |
| title_short | Development of strategies for genetic manipulation and fine‐tuning of a chloroplast retrograde signal 3′‐phosphoadenosine 5′‐phosphate |
| title_sort | development of strategies for genetic manipulation and fine tuning of a chloroplast retrograde signal 3 phosphoadenosine 5 phosphate |
| topic | 3′‐phosphoadenosine‐5′‐phosphate drought gene silencing retrograde signaling RNAi SAL1 |
| url | https://doi.org/10.1002/pld3.31 |
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