Soilborne bacterium Klebsiella pneumoniae promotes cluster root formation in white lupin through ethylene mediation.

Cluster roots of white lupin are induced by low phosphorus (LP) to efficiently access unavailable P, but how soilborne microbes are associated with cluster root formation (CRF) is unclear. We investigated the roles of soilborne bacteria in CRF response to LP by high-throughput sequencing and root-bacteria interactions. Cluster root number was significantly decreased in plants grown in sterilized soil compared with nonsterilized soil. Proteobacteria was enriched in CR, as shown by microbiome analysis of soil (bulk, rhizosphere, and rhizosheath) and roots (main, lateral, and CR). Large-scale gene expression level implicated ethylene mediation in CRF. Klebsiella pneumoniae (P7), a soilborne bacterium belonging to Proteobacteria, was isolated from CR. Among 11 isolated strains, P7 exhibited the highest 1-aminocyclopropane-1-carboxylate deaminase (ACCD) activity; this enzyme inhibits the biosynthesis of ethylene in plants by the cleavage of the ethylene precursor 1-aminocyclopropane-1-carboxylic acid and promotes CRF under LP. We constructed an ACCD-deficit mutant accd in the P7 genetic background. The loss-of-function mutation failed to promote CRF under LP conditions. Also, auxin responses may be involved in K. pneumoniae-ethylene-mediated CRF. Overall, we propose that the soilborne bacterium K. pneumoniae promotes CRF of white lupin in response to LP by ethylene mediation.

Root-Bacteria Associations Boost Rhizosheath Formation in Moderately Dry Soil through Ethylene Responses.

The rhizosheath is a layer of soil around the root that provides a favorable environment for soil microbe enrichment and root growth. Rice (Oryza sativa) roots form rhizosheaths under moderate soil drying (MSD) conditions, but how the rhizosheath forms associations with microbes is unclear. To investigate rice rhizosheath formation under MSD, we employed a multiphasic approach, integrating data from high-throughput sequencing and root-bacteria interactions. Rice roots formed a pronounced rhizosheath under MSD, but not under continuous flooding regimens. Plant growth-promoting rhizobacteria of the Enterobacteriaceae were enriched in rhizosheaths of two different rice varieties, 'Gaoshan 1' (drought tolerant) and 'Nipponbare' (drought sensitive). RNA-sequencing analysis revealed that the ethylene pathway was induced in the rhizosheath-root system under MSD. Enterobacter aerogenes, a bacterium isolated from the rhizosheath, degrades the ethylene precursor 1-aminocyclopropane-1-carboxylate, thereby increasing rhizosheath formation. Furthermore, a 1-aminocyclopropane-1-carboxylate deaminase-deficient mutant of E. aerogenes failed to enhance rice rhizosheath formation. Our results suggest that root-bacteria associations substantially contribute to rhizosheath formation in rice under MSD conditions by mechanisms that involve the ethylene response. These data inform strategies to reduce water consumption in rice production, one of the most water-intensive human activities.

Abscisic acid mediates barley rhizosheath formation under mild soil drying by promoting root hair growth and auxin response.

Soil drying enhances root ABA accumulation and rhizosheath formation, but whether ABA mediates rhizosheath formation is unclear. Here, we used the ABA-deficient mutant Az34 to investigate molecular and morphological changes by which ABA could affect rhizosheath formation. Mild soil drying with intermittent watering increased rhizosheath formation by promoting root and root hair elongation. Attenuated root ABA accumulation in Az34 barley constrained the promotion of root length and root hair length by drying soil, such that Az34 had a smaller rhizosheath. Pharmacological experiments of adding fluridone (an ABA biosynthesis inhibitor) and ABA to drying soil restricted and enhanced rhizosheath formation respectively in Az34 and wild-type Steptoe barley. RNA sequencing suggested that ABA accumulation mediates auxin synthesis and responses and root and root hair elongation in drying soil. In addition, adding indole-3-acetic acid (IAA) to drying soil increased rhizosheath formation by promoting root and root hair elongation in Steptoe and Az34 barley. Together, these results show that ABA accumulation induced by mild soil drying enhance barley rhizosheath formation, which may be achieved through promoting auxin response.

Coordination of root auxin with the fungus Piriformospora indica and bacterium Bacillus cereus enhances rice rhizosheath formation under soil drying.

Moderate soil drying (MSD) is a promising agricultural technique that can reduce water consumption and enhance rhizosheath formation promoting drought resistance in plants. The endophytic fungus Piriformospora indica (P. indica) with high auxin production may be beneficial for rhizosheath formation. However, the integrated role of P. indica with native soil microbiome in rhizosheath formation is unclear. Here, we investigated the roles of P. indica and native bacteria on rice rhizosheath formation under MSD using high-throughput sequencing and rice mutants. Under MSD, rice rhizosheath formation was significantly increased by around 30% with P. indica inoculation. Auxins in rice roots and P. indica were responsible for the rhizosheath formation under MSD. Next, the abundance of the genus Bacillus, known as plant growth-promoting rhizobacteria, was enriched in the rice rhizosheath and root endosphere with P. indica inoculation under MSD. Moreover, the abundance of Bacillus cereus (B. cereus) with high auxin production was further increased by P. indica inoculation. After inoculation with both P. indica and B. cereus, rhizosheath formation in wild-type or auxin efflux carrier OsPIN2 complemented line rice was higher than that of the ospin2 mutant. Together, our results suggest that the interaction of the endophytic fungus P. indica with the native soil bacterium B. cereus favors rice rhizosheath formation by auxins modulation in rice and microbes under MSD. This finding reveals a cooperative contribution of P. indica and native microbiota in rice rhizosheath formation under moderate soil drying, which is important for improving water use in agriculture.

Rhizosheath microbes induce root immune response under soil drying.

The rhizosheath is an important drought-adaptive trait in roots of many angiosperms and has been regarded as a potential trait for future agricultural sustainability. In recent studies, we found that rice roots could form a pronounced rhizosheath under moderate soil drying (MSD) but not under continuous flooding irrigation (CF). The formation of rhizosheaths substantially changes the microbial community structure in endosphere root tissues and the rhizosphere in rice, which may induce a plant immune response. However, the manner by which the formation of rhizosheaths regulates the immune system of roots remains largely unknown. Here, we have analyzed the root transcriptomes of drought-tolerant rice and drought-sensitive rice under both MSD (rhizosheath-root) and CF (root without rhizosheath) conditions. Our results suggest that rhizosheath-associated microbes may trigger plant immune pathways in root under MSD, including the first line of defense component pattern-triggered immunity and the second line of defense component effector-triggered immunity. These data expand our understanding of rhizosheath-associated microbes and plant interactions.