Abstract
Abstract Phosphate transporters (PHTs) are H 2 PO 4 ⁻ /H + transporters that facilitate inorganic phosphate uptake from the soil and its distribution within plants. In citrus, seasonal summer drought constrains yield and limits phosphate uptake. However, the roles of PHT genes in drought response remain poorly understood. In this study, we identified 20 PtPHTs in the Poncirus trifoliata genome and classified them into five subgroups (I to V) based on phylogenetic relationships. Promoter analysis revealed abundant cis- acting elements associated with abiotic stress and phytohormone responses, while synteny analysis indicated that gene duplication contributed to PtPHT family expansion. Expression profile showed that several genes, including PtPHT1;1 , PtPHT1;2 , PtPHT1;3 , PtPHT3;3 , PtPHT4;3 , PtPHT5;1 , and PtPHT5;2 were significantly upregulated by drought, with PtPHT1;2 exhibiting the strongest response (about 16.4 folds). Silencing PtPHT1;2 markedly reduced phosphorus (P) accumulation in transgenic citrus calli, confirming its functional role in inorganic phosphate uptake. Yeast one-hybrid (Y1H) screening identified Related to AP2.12 (PtRap2.12), an APETALA2/ETHYLENE RESPONSE FACTOR (AP2/ERF) transcription factor, as an upstream regulator of PtPHT1;2 . This interaction was validated by Y1H and dual-luciferase assays, demonstrating that PtRap2.12 directly binds to and activates the PtPHT1;2 promoter. Functional analysis further showed that overexpression of PtRap2.12 enhances drought tolerance of transgenic plants, as indicated by improved growth, reduced membrane damage, elevated antioxidant enzyme activities, and increased P accumulation. Moreover, transgenic plants displayed upregulated expression of NtPHT1;1 in shoots and NtPHT1;4 in roots of tobacco ( Nicotiana tabacum ) under drought conditions. In summary, this study provides a comprehensive analysis of PtPHT genes in P. trifoliata and highlights the role of the PtRap2.12– PtPHT1;2 module in regulating both phosphate uptake and drought tolerance, offering a theoretical basis for breeding citrus rootstocks with enhanced resistance to drought and phosphate deficiency.
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Claims 14
Claim · model-written summary
Twenty PtPHT genes were identified and classified into five subgroups.
- Taxon
- Poncirus trifoliata
Citrus trifoliatae0 - Gene
- PtPHTse0
- Outcome
- classified them into five subgroupse0
- Value
- 20 PtPHTse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“identified 20 PtPHTs in the Poncirus trifoliata genome and classified them into five subgroups (I to V)”
Claim · model-written summary
Promoters had abundant stress- and hormone-related cis elements.
- Taxon
- Poncirus trifoliataCitrus trifoliatat0
- Outcome
- abundant cis- acting elementse0
- Qualifier
- abiotic stress and phytohormone responsese0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Promoter analysis revealed abundant cis- acting elements associated with abiotic stress and phytohormone responses”
t0“Poncirus trifoliata”
Claim · model-written summary
Gene duplication contributed to PtPHT family expansion.
- Taxon
- Poncirus trifoliataCitrus trifoliatat0
- Gene
- PtPHTe0
- Outcome
- PtPHT family expansione0
- Qualifier
- gene duplicatione0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“synteny analysis indicated that gene duplication contributed to PtPHT family expansion”
t0“Poncirus trifoliata”
Claim · model-written summary
Seven named PtPHT genes were significantly upregulated by drought.
- Taxon
- Poncirus trifoliataCitrus trifoliatat0
- Gene
- PtPHT1;1 , PtPHT1;2 , PtPHT1;3 , PtPHT3;3 , PtPHT4;3 , PtPHT5;1 , and PtPHT5;2e0
- Outcome
- upregulated by droughte0
- Direction
- upregulatede0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“PtPHT1;1 , PtPHT1;2 , PtPHT1;3 , PtPHT3;3 , PtPHT4;3 , PtPHT5;1 , and PtPHT5;2 were significantly upregulated by drought”
t0“Poncirus trifoliata”
Claim · model-written summary
PtPHT1;2 showed the strongest drought expression response, about 16.4 folds.
- Taxon
- Poncirus trifoliataCitrus trifoliatat0
- Gene
- PtPHT1;2e0
- Outcome
- responsee0
- Direction
- strongeste0
- Value
- about 16.4 foldse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“PtPHT1;2 exhibiting the strongest response (about 16.4 folds)”
t0“Poncirus trifoliata”
Claim · model-written summary
Silencing PtPHT1;2 reduced phosphorus accumulation in citrus calli.
- Organ / stage
- callie0
- Treatment
- Silencing PtPHT1;2e0
- Gene
- PtPHT1;2e0
- Outcome
- phosphorus (P) accumulatione0
- Direction
- reducede0
CitrusNot reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Silencing PtPHT1;2 markedly reduced phosphorus (P) accumulation in transgenic citrus calli”
Claim · model-written summary
PtRap2.12 was identified as an upstream regulator of PtPHT1;2.
- Taxon
- Poncirus trifoliataCitrus trifoliatat0
- Gene
- Related to AP2.12 (PtRap2.12)e0
- Outcome
- upstream regulator of PtPHT1;2e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“identified Related to AP2.12 (PtRap2.12), an APETALA2/ETHYLENE RESPONSE FACTOR (AP2/ERF) transcription factor, as an upstream regulator of PtPHT1;2”
t0“Poncirus trifoliata”
Claim · model-written summary
PtRap2.12 directly binds and activates the PtPHT1;2 promoter.
- Taxon
- Poncirus trifoliataCitrus trifoliatat0
- Gene
- PtRap2.12e0
- Outcome
- activates the PtPHT1;2 promotere0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“PtRap2.12 directly binds to and activates the PtPHT1;2 promoter”
t0“Poncirus trifoliata”
Claim · model-written summary
PtRap2.12 overexpression improved growth and drought tolerance.
- Taxon
- Poncirus trifoliataCitrus trifoliatat0
- Treatment
- overexpression of PtRap2.12e0
- Gene
- PtRap2.12e0
- Outcome
- drought tolerancee0
- Direction
- enhancese0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“overexpression of PtRap2.12 enhances drought tolerance of transgenic plants, as indicated by improved growth”
t0“Poncirus trifoliata”
Claim · model-written summary
PtRap2.12 overexpression reduced membrane damage.
- Taxon
- Poncirus trifoliataCitrus trifoliatat0
- Treatment
- overexpression of PtRap2.12e0
- Outcome
- membrane damagee0
- Direction
- reducede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“overexpression of PtRap2.12 enhances drought tolerance of transgenic plants, as indicated by improved growth, reduced membrane damage”
t0“Poncirus trifoliata”
Claim · model-written summary
PtRap2.12 overexpression elevated antioxidant enzyme activities.
- Taxon
- Poncirus trifoliataCitrus trifoliatat0
- Treatment
- overexpression of PtRap2.12e0
- Outcome
- antioxidant enzyme activitiese0
- Direction
- elevatede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“overexpression of PtRap2.12 enhances drought tolerance of transgenic plants, as indicated by improved growth, reduced membrane damage, elevated antioxidant enzyme activities”
t0“Poncirus trifoliata”
Claim · model-written summary
PtRap2.12 overexpression increased phosphorus accumulation.
- Taxon
- Poncirus trifoliataCitrus trifoliatat0
- Treatment
- overexpression of PtRap2.12e0
- Outcome
- P accumulatione0
- Direction
- increasede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“overexpression of PtRap2.12 enhances drought tolerance of transgenic plants, as indicated by improved growth, reduced membrane damage, elevated antioxidant enzyme activities, and increased P accumulation”
t0“Poncirus trifoliata”
Claim · model-written summary
Transgenic tobacco shoots upregulated NtPHT1;1 under drought.
- Taxon
- tobacco ( Nicotiana tabacum )
Nicotiana tabacume0 - Organ / stage
- shootse0
- Gene
- NtPHT1;1e0
- Outcome
- expression of NtPHT1;1e0
- Direction
- upregulatede0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“upregulated expression of NtPHT1;1 in shoots and NtPHT1;4 in roots of tobacco ( Nicotiana tabacum ) under drought conditions”
Claim · model-written summary
Transgenic tobacco roots upregulated NtPHT1;4 under drought.
- Taxon
- tobacco ( Nicotiana tabacum )Nicotiana tabacume0
- Organ / stage
- rootse0
- Gene
- NtPHT1;4e0
- Outcome
- NtPHT1;4e0
- Direction
- upregulatede0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“upregulated expression of NtPHT1;1 in shoots and NtPHT1;4 in roots of tobacco ( Nicotiana tabacum ) under drought conditions”
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