Abstract
Cross-kingdom RNA interference has emerged as an important mechanism in plant-pathogen interactions, yet how fungal small RNAs coordinate host immune suppression remains unclear. Here, we demonstrate that Alternaria alternata f. sp. mali, the causal agent of apple leaf spot, suggests microRNA-like RNAs (milRNAs) into host tissues in association with extracellular vesicles (EVs). Small RNA sequencing of fungal EVs identified multiple milRNAs, among which AamilR251, AamilR292 and AamilR004 were the three most abundant species. These AamilRNAs target three apple (Malus domestica) immune-related kinases (MdMAPKKK1, MdRLK2 and MdMAPKK6). Molecular and genetic analyses revealed that these proteins form an interconnected signalling module and a linear phosphorylation cascade (MdRLK2-MdCRK10-MdMAPKKK1-MdMAPKK6) required for effective defence activation. Deletion of individual AamilRNAs significantly attenuated fungal virulence and restored expression of their respective host targets. Conversely, overexpression of MdMAPKKK1, MdRLK2 or MdMAPKK6 enhanced resistance to infection. Pull-down assays identified MdCRK10 as an interacting partner of MdRLK2, forming a complex with MAPKs that activates defence signalling. MdRLK2 phosphorylates MdCRK10 at S144 and S363, and mutation of these sites disrupts signal transduction and compromises disease resistance. These findings reveal a central immune signalling network in apple and identify key components for improving resistance against fungal pathogens.
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Claims 13
Claim · model-written summary
Alternaria alternata f. sp. mali milRNAs were associated with extracellular vesicles in host tissues.
- Taxon
- apple
Malus domesticae0 - Interactor
- Alternaria alternata f. sp. mali
Alternaria alternata f. sp. malie0pathogen - Outcome
- microRNA-like RNAs (milRNAs) into host tissuese0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Alternaria alternata f. sp. mali, the causal agent of apple leaf spot, suggests microRNA-like RNAs (milRNAs) into host tissues in association with extracellular vesicles (EVs).”
Claim · model-written summary
AamilR251, AamilR292, and AamilR004 were the three most abundant fungal EV milRNAs.
- Taxon
- appleMalus domesticae1
- Interactor
- Alternaria alternata f. sp. maliAlternaria alternata f. sp. malie2pathogen
- Gene
- AamilR251, AamilR292 and AamilR004e0
- Outcome
- the three most abundant speciese0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Small RNA sequencing of fungal EVs identified multiple milRNAs, among which AamilR251, AamilR292 and AamilR004 were the three most abundant species.”
e1“These AamilRNAs target three apple (Malus domestica) immune-related kinases (MdMAPKKK1, MdRLK2 and MdMAPKK6).”
e2“Alternaria alternata f. sp. mali, the causal agent of apple leaf spot, suggests microRNA-like RNAs (milRNAs) into host tissues in association with extracellular vesicles (EVs).”
Claim · model-written summary
The fungal AamilRNAs target three apple immune-related kinases.
- Taxon
- appleMalus domesticae0
- Interactor
- Alternaria alternata f. sp. maliAlternaria alternata f. sp. malie2pathogen
- Gene
- MdMAPKKK1, MdRLK2 and MdMAPKK6e0
- Outcome
- immune-related kinasese0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“These AamilRNAs target three apple (Malus domestica) immune-related kinases (MdMAPKKK1, MdRLK2 and MdMAPKK6).”
e1“These AamilRNAs target three apple (Malus domestica) immune-related kinases (MdMAPKKK1, MdRLK2 and MdMAPKK6).”
e2“Alternaria alternata f. sp. mali, the causal agent of apple leaf spot, suggests microRNA-like RNAs (milRNAs) into host tissues in association with extracellular vesicles (EVs).”
Claim · model-written summary
The apple proteins form an interconnected signaling module and a phosphorylation cascade required for defense activation.
- Taxon
- appleMalus domesticae1
- Gene
- MdRLK2-MdCRK10-MdMAPKKK1-MdMAPKK6e0
- Outcome
- effective defence activatione0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“these proteins form an interconnected signalling module and a linear phosphorylation cascade (MdRLK2-MdCRK10-MdMAPKKK1-MdMAPKK6) required for effective defence activation.”
e1“These AamilRNAs target three apple (Malus domestica) immune-related kinases (MdMAPKKK1, MdRLK2 and MdMAPKK6).”
e2“Alternaria alternata f. sp. mali, the causal agent of apple leaf spot, suggests microRNA-like RNAs (milRNAs) into host tissues in association with extracellular vesicles (EVs).”
Claim · model-written summary
Deleting individual AamilRNAs significantly attenuated fungal virulence.
- Taxon
- appleMalus domesticae1
- Treatment
- Deletion of individual AamilRNAse0
- Interactor
- Alternaria alternata f. sp. maliAlternaria alternata f. sp. malie2pathogen
- Outcome
- fungal virulencee0
- Direction
- attenuatede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Deletion of individual AamilRNAs significantly attenuated fungal virulence and restored expression of their respective host targets.”
e1“These AamilRNAs target three apple (Malus domestica) immune-related kinases (MdMAPKKK1, MdRLK2 and MdMAPKK6).”
e2“Alternaria alternata f. sp. mali, the causal agent of apple leaf spot, suggests microRNA-like RNAs (milRNAs) into host tissues in association with extracellular vesicles (EVs).”
Claim · model-written summary
Deleting individual AamilRNAs restored expression of their host targets.
- Taxon
- appleMalus domesticae1
- Treatment
- Deletion of individual AamilRNAse0
- Outcome
- expression of their respective host targetse0
- Direction
- restorede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Deletion of individual AamilRNAs significantly attenuated fungal virulence and restored expression of their respective host targets.”
e1“These AamilRNAs target three apple (Malus domestica) immune-related kinases (MdMAPKKK1, MdRLK2 and MdMAPKK6).”
e2“Alternaria alternata f. sp. mali, the causal agent of apple leaf spot, suggests microRNA-like RNAs (milRNAs) into host tissues in association with extracellular vesicles (EVs).”
Claim · model-written summary
Overexpressing MdMAPKKK1 increased resistance to infection.
- Taxon
- appleMalus domesticae1
- Treatment
- overexpression of MdMAPKKK1e0
- Gene
- MdMAPKKK1e0
- Outcome
- resistance to infectione0
- Direction
- enhancede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“overexpression of MdMAPKKK1, MdRLK2 or MdMAPKK6 enhanced resistance to infection.”
e1“These AamilRNAs target three apple (Malus domestica) immune-related kinases (MdMAPKKK1, MdRLK2 and MdMAPKK6).”
e2“Alternaria alternata f. sp. mali, the causal agent of apple leaf spot, suggests microRNA-like RNAs (milRNAs) into host tissues in association with extracellular vesicles (EVs).”
Claim · model-written summary
Overexpressing MdRLK2 increased resistance to infection.
- Taxon
- appleMalus domesticae1
- Treatment
- overexpression of MdMAPKKK1, MdRLK2 or MdMAPKK6e0
- Gene
- MdRLK2e0
- Outcome
- resistance to infectione0
- Direction
- enhancede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“overexpression of MdMAPKKK1, MdRLK2 or MdMAPKK6 enhanced resistance to infection.”
e1“These AamilRNAs target three apple (Malus domestica) immune-related kinases (MdMAPKKK1, MdRLK2 and MdMAPKK6).”
e2“Alternaria alternata f. sp. mali, the causal agent of apple leaf spot, suggests microRNA-like RNAs (milRNAs) into host tissues in association with extracellular vesicles (EVs).”
Claim · model-written summary
Overexpressing MdMAPKK6 increased resistance to infection.
- Taxon
- appleMalus domesticae1
- Treatment
- overexpression of MdMAPKKK1, MdRLK2 or MdMAPKK6e0
- Gene
- MdMAPKK6e0
- Outcome
- resistance to infectione0
- Direction
- enhancede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“overexpression of MdMAPKKK1, MdRLK2 or MdMAPKK6 enhanced resistance to infection.”
e1“These AamilRNAs target three apple (Malus domestica) immune-related kinases (MdMAPKKK1, MdRLK2 and MdMAPKK6).”
e2“Alternaria alternata f. sp. mali, the causal agent of apple leaf spot, suggests microRNA-like RNAs (milRNAs) into host tissues in association with extracellular vesicles (EVs).”
Claim · model-written summary
MdCRK10 interacts with MdRLK2 in a complex with MAPKs that activates defense signaling.
- Taxon
- appleMalus domesticae1
- Gene
- MdCRK10e0
- Outcome
- activates defence signallinge0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Pull-down assays identified MdCRK10 as an interacting partner of MdRLK2, forming a complex with MAPKs that activates defence signalling.”
e1“These AamilRNAs target three apple (Malus domestica) immune-related kinases (MdMAPKKK1, MdRLK2 and MdMAPKK6).”
e2“Alternaria alternata f. sp. mali, the causal agent of apple leaf spot, suggests microRNA-like RNAs (milRNAs) into host tissues in association with extracellular vesicles (EVs).”
Claim · model-written summary
MdRLK2 phosphorylates MdCRK10 at S144 and S363.
- Taxon
- appleMalus domesticae1
- Gene
- MdRLK2e0
- Outcome
- phosphorylates MdCRK10 at S144 and S363e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“MdRLK2 phosphorylates MdCRK10 at S144 and S363”
e1“These AamilRNAs target three apple (Malus domestica) immune-related kinases (MdMAPKKK1, MdRLK2 and MdMAPKK6).”
e2“Alternaria alternata f. sp. mali, the causal agent of apple leaf spot, suggests microRNA-like RNAs (milRNAs) into host tissues in association with extracellular vesicles (EVs).”
Claim · model-written summary
Mutating MdCRK10 phosphorylation sites disrupted signal transduction.
- Taxon
- appleMalus domesticae1
- Treatment
- mutation of these sitese0
- Outcome
- signal transductione0
- Direction
- disruptse0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“mutation of these sites disrupts signal transduction and compromises disease resistance.”
e1“These AamilRNAs target three apple (Malus domestica) immune-related kinases (MdMAPKKK1, MdRLK2 and MdMAPKK6).”
e2“Alternaria alternata f. sp. mali, the causal agent of apple leaf spot, suggests microRNA-like RNAs (milRNAs) into host tissues in association with extracellular vesicles (EVs).”
Claim · model-written summary
Mutating MdCRK10 phosphorylation sites compromised disease resistance.
- Taxon
- appleMalus domesticae1
- Treatment
- mutation of these sitese0
- Outcome
- disease resistancee0
- Direction
- compromisese0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“mutation of these sites disrupts signal transduction and compromises disease resistance.”
e1“These AamilRNAs target three apple (Malus domestica) immune-related kinases (MdMAPKKK1, MdRLK2 and MdMAPKK6).”
e2“Alternaria alternata f. sp. mali, the causal agent of apple leaf spot, suggests microRNA-like RNAs (milRNAs) into host tissues in association with extracellular vesicles (EVs).”
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See all 21 papers →Claims are grouped by organism, treatment and what was measured or targeted. A paper counts as positive when any of its claims in the group is positive.