Lights off - Role of bioluminescence for the biology of the biocontrol agent Photorhabdus luminescens.

Bioluminescence is found across various organisms having crucial functions for biotic interactions and stress adaptation. The only known terrestrial bioluminescent bacteria are entomopathogenic bacteria of the genus Photorhabdus. However, the reason why these bacteria produce light is not understood. P. luminescens exists in two cell forms called primary (1°) and secondary (2°) cells. The 1° cells colonize the nematode symbiosis partner and produce bright light, whereas 2° cells colonize plant roots only emitting weak light. Here we show that bioluminescence is important but not essential for the biology of the bacteria. Deletion of the luxCDABE operon in 1° cells impaired insect pathogenicity and nematode interaction. The complete loss of light of 2° cells resulted in enhanced plant root colonization, enhanced haemolysis, and reduced oxidative stress adaptation. Since bioluminescence is not essential for the survival of the bacteria, P. luminescens Δlux 1° and 2° emerged as useful tools for bioluminescence-based reporter assays.

Secondary metabolites in host pears defense against two fruit borers and cytochrome-P450-mediated counter-defense.

Herbivorous insects have evolved metabolic strategies to survive the challenges posed by plant secondary metabolites (SMs). This study reports an exploration of SMs present in pears, which serve as a defense against invasive Cydia pomonella and native Grapholita molesta and their counter-defense response. The feeding preferences of fruit borers are influenced by the softening of two pear varieties as they ripen. The content of SMs, such as quercetin and rutin, increases due to feeding by fruit borers. Notably, quercetin levels only increase after C. pomonella feeding. The consumption of SMs affects the growth of fruit borer population differently, potentially due to the activation of P450 genes by SMs. These two fruit borers are equipped with specific P450 enzymes that specialize in metabolizing quercetin and rutin, enabling them to adapt to these SMs in their host fruits. These findings provide valuable insights into the coevolution of plants and herbivorous insects.

Inhibition of a conserved bacterial dual-specificity phosphatase confers plant tolerance to Candidatus Liberibacter spp.

"Candidatus Liberibacter spp." are insect-vectored, fastidious, and vascular-limited phytopathogens. They are the presumptive causal agents of potato zebra chip, tomato vein clearing, and the devastating citrus greening disease worldwide. There is an urgent need to develop new strategies to control them. In this study, we characterized a dual-specificity serine/tyrosine phosphatase (STP) that is well conserved among thirty-three geographically diverse "Candidatus Liberibacter spp." and strains that infect multiple Solanaceaea and citrus spp. The STP is expressed in infected plant tissues, localized at the plant cytosol and plasma membrane, and interferes with plant cell death responses. We employed an in silico target-based molecular modeling and ligand screen to identify two small molecules with high binding affinity to STP. Efficacy studies demonstrated that the two molecules can inhibit "Candidatus Liberibacter spp." but not unrelated pathogens and confer plant disease tolerance. The inhibitors and strategies are promising means to control "Candidatus Liberibacter spp."

Role of PMR4 and PDLP1 in priming of early acting penetration defense by resistance-inducing ß-amino acids.

R-ß-homoserine (RBH) and ß-aminobutyric acid (BABA) induce resistance against the oomycete Hyaloperonospora arabidopsidis (Hpa) in Arabidopsis, which is based on priming of multiple defense layers, including early acting penetration resistance at the cell wall. Here, we have examined the molecular basis of RBH- and BABA-primed defense by cell wall papillae against Hpa. Three-dimensional reconstruction of Hpa-induced papillae by confocal microscopy revealed no structural differences between control-, RBH-, and BABA-treated plants after Hpa challenge. However, mutations affecting POWDERY MILDEW RESISTANCE 4 or PLASMODESMATA LOCATED PROTEINs (PDLPs) only impaired BABA-induced penetration resistance and not RBH-induced penetration resistance. Furthermore, PDLP1 over-expression mimicked primed penetration resistance, while the intensity of GFP-tagged PDLP1 at germinating Hpa conidiospores was increased in BABA-primed plants but not RBH-primed plants. Our study reveals new regulatory layers of immune priming by ß-amino acids and supports the notion that penetration resistance is a multifaceted defense layer that can be achieved through seperate pathways.

Comparative genomics analysis reveals genetic characteristics and nitrogen fixation profile of Bradyrhizobium.

Bradyrhizobium is a genus of nitrogen-fixing bacteria, with some species producing nodules in leguminous plants. Investigations into Bradyrhizobium have recently revealed its substantial genetic resources and agricultural benefits, but a comprehensive survey of its genetic diversity and functional properties is lacking. Using a panel of various strains (N = 278), this study performed a comparative genomics analysis to anticipate genes linked with symbiotic nitrogen fixation. Bradyrhizobium's pan-genome consisted of 84,078 gene families, containing 824 core genes and 42,409 accessory genes. Core genes were mainly involved in crucial cell processes, while accessory genes served diverse functions, including nitrogen fixation and nodulation. Three distinct genetic profiles were identified based on the presence/absence of gene clusters related to nodulation, nitrogen fixation, and secretion systems. Most Bradyrhizobium strains from soil and non-leguminous plants lacked major nif/nod genes and were evolutionarily more closely related. These findings shed light on Bradyrhizobium's genetic features for symbiotic nitrogen fixation.

Genome-scale cis-acting catabolite-responsive element editing confers Bacillus pumilus LG3145 plant-beneficial functions.

Rhizosphere dwelling microorganism such as Bacillus spp. are helpful for crop growth. However, these functions are adversely affected by long-term synthetic fertilizer application. We developed a modified CRISPR/Cas9 system using non-specific single-guide RNAs to disrupt the genome-wide cis-acting catabolite-responsive elements (cres) in a wild-type Bacillus pumilus strain, which conferred dual plant-benefit properties. Most of the mutations occurred around imperfectly matched cis-acting elements (cre-like sites) in genes that are mainly involved in carbon and secondary metabolism pathways. The comparative metabolomics and transcriptome results revealed that carbon is likely transferred to some pigments, such as riboflavin, carotenoid, and lycopene, or non-ribosomal peptides, such as siderophore, surfactin, myxochelin, and bacilysin, through the pentose phosphate and amino acid metabolism pathways. Collectively, these findings suggested that the mutation of global cre-like sequences in the genome might alter carbon flow, thereby allowing beneficial biological interactions between the rhizobacteria and plants.

Plants as vectors for environmental prion transmission.

Prions cause fatal neurodegenerative diseases and exhibit remarkable durability, which engenders a wide array of potential exposure scenarios. In chronic wasting disease of deer, elk, moose, and reindeer and in scrapie of sheep and goats, prions are transmitted via environmental routes and the ability of plants to accumulate and subsequently transmit prions has been hypothesized, but not previously demonstrated. Here, we establish the ability of several crop and other plant species to take up prions via their roots and translocate them to above-ground tissues from various growth media including soils. We demonstrate that plants can accumulate prions in above-ground tissues to levels sufficient to transmit disease after oral ingestion by mice. Our results suggest plants may serve as vectors for prion transmission in the environment-a finding with implications for wildlife conservation, agriculture, and public health.

Bumblebees negotiate a trade-off between nectar quality and floral biomechanics.

How and why pollinators choose which flowers to visit are fundamental, multifaceted questions in pollination biology, yet most studies of floral traits measure simple relative preferences. Here, we used vertically and horizontally oriented slippery-surfaced artificial flowers to test whether bumblebees could make a trade-off between floral handling difficulty and nectar sucrose concentration. We quantified foraging energetics, thereby resolving the rationale behind the bees' foraging decisions. The bees chose flowers with either a high handling cost or low sucrose concentration, depending on which was the energetically favorable option. Their behavior agreed with the critical currency being the rate of energy return (net energy collected per unit time), not energetic efficiency (net energy collected per unit energy spent). This suggests that bumblebees prioritize immediate carbohydrate flow to the nest rather than energy gain over the working lifespan of each bee. Trade-off paradigms like these are a powerful approach for quantifying pollinator trait preferences.

The PtdIns3P phosphatase MtMP promotes symbiotic nitrogen fixation via mitophagy in Medicago truncatula.

Symbiotic nitrogen fixation is a complex process in which legumes interact with rhizobia under nitrogen starvation. In this study, we found that myotubularin phosphatase (MtMP) is mainly expressed in roots and nodules in Medicago truncatula. MtMP promotes autophagy by dephosphorylating PtdIns3P on autophagosomes. The mp mutants inoculated with rhizobia showed a significant reduction in nitrogenase activity and significantly higher number of mitochondria than those of wild-type plants under nitrogen starvation, indicating that MtMP is involved in mitophagy of the infection zone. Mitophagy may provide carbon skeletons and nitrogen for the development of bacteroids and the reprogramming of infected cells. In conclusion, we found, for the first time, that myotubularin phosphatase is involved in autophagy in plants. MtMP-involved autophagy plays an active role in symbiotic nitrogen fixation. These results deepen our understanding of symbiotic nitrogen fixation.

Host-plant adaptation in xylophagous insect-microbiome systems: Contributionsof longicorns and gut symbionts revealed by parallel metatranscriptome.

Adaptation to host plants is of great significance in the ecology of xylophagous insects. The specific adaptation to woody tissues is made possible through microbial symbionts. We investigated the potential roles of detoxification, lignocellulose degradation, and nutrient supplementation of Monochamus saltuarius and its gut symbionts in host plant adaptation using metatranscriptome. The gut microbial community structure of M. saltuarius that fed on the two plant species were found to be different. Plant compound detoxification and lignocellulose degradation genes have been identified in both beetles and gut symbionts. Most differentially expressed genes associated with host plant adaptations were up-regulated in larvae fed on the less suitable host (Pinus tabuliformis) compared to larvae fed on the suitable host (Pinus koraiensis). Our findings indicated that M. saltuarius and its gut microbes respond to plant secondary substances through systematic transcriptome responses, allowing them to adapt to unsuitable host plants.

Enhancing biocontrol potential of Trichogramma chilonis against borer pests of wheat and chickpea.

Communication between plants and herbivores occur mostly through chemicals. Plants emit volatiles in response to the attack of herbivores called herbivore-induced plant volatiles (HIPVs), which are employed by the plants to attract their herbivores' natural enemies. Promising HIPVs when used in the form of controlled release formulations under field conditions can act as arrestants of released or wild population of parasitoids to spend comparatively more time in searching of various stages of herbivores; thus management of crop pests is enhanced. Gel formulation of octadecane (Saturated hydrocarbon) when applied 24 h after release of Trichogramma chilonis in wheat and chickpea enhanced the foraging activity against pink stem borer (Sesamia inferens) and pod borer (Helicoverpa armigera), respectively resulting in reduced symptoms and increased grain yields. Controlled release formulations of promising plant volatile organic compounds (pVOCs) can be useful to augment and enhance foraging activity of natural enemies against crop herbivores.

Microbial biogeochemical cycling reveals the sustainability of the rice-crayfish co-culture model.

Aquaculture has great potential in nourishing the global growing population, while such staggering yields are coupled with environmental pollution. Rice-crayfish co-culture models (RCFP) have been widely adopted in China due to their eco-friendliness. However, little is known about RCFP's microbiome pattern, which hinders our understanding of its sustainability. This study has conducted metagenomic analysis across aquaculture models and habitats, which revealed aquaculture model-specific biogeochemical cycling pattern (e.g., nitrogen (N), sulfur (S), and carbon (C)): RCFP is advantageous in N-assimilation, N-contamination, and S-pollutants removal, while non-RCFP features N denitrification process and higher S metabolism ability, producing several hazardous pollutants in non-RCFP (e.g., nitric oxide, nitrogen monoxide, and sulfide). Moreover, RCFP has greater capacity for carbohydrate enzyme metabolism compared with non-RCFP in environmental habitats, but not in crayfish gut. Collectively, RCFP plays an indispensable role in balancing aquaculture productivity and environmental protection, which might be applied to the blue transformation of aquaculture.

Refuges of conventional host plants counter dominant resistance of cotton bollworm to transgenic Bt cotton.

Transgenic crops have revolutionized insect pest control, but evolution of resistance by pests threatens their continued success. The primary strategy for combating pest resistance to crops producing insecticidal proteins from Bacillus thuringiensis (Bt) uses refuges of non-Bt host plants to allow survival of susceptible insects. The prevailing paradigm is that refuges delay resistance that is rare and recessively inherited. However, we discovered refuges countered resistance to Bt cotton that was neither rare nor recessive. In a 15-year field study of the cotton bollworm, the frequency of a mutation conferring dominant resistance to Bt cotton surged 100-fold from 2006 to 2016 yet did not rise from 2016 to 2020. Computer simulations indicate the increased refuge percentage from 2016 to 2020 is sufficient to explain the observed halt in the evolution of resistance. The results also demonstrate the efficacy of a Bt crop can be sustained by non-Bt refuges of other crops.

Transcriptomic investigation reveals a physiological mechanism for Beauveria bassiana to survive under linoleic acid stress.

In integrated pest management program (IPM), the compatibility of mycoinsecticides with bioactive fungicides [e.g., unsaturated fatty acids (UFAs)] has attracted more and more attention; however, the mechanisms underlying fungal resistance to UFAs remain largely unknown. In this study, Beauveria bassiana, an entomopathogenic fungus, was used to explore fungal responses to linoleic acid (LA). Genome-wide expression revealed the transcriptomic responses of fungal cells to LA in a stress-intensity-dependent manner. Enrichment analyses indicated that the up-regulated differentially expressed genes (DEGs) are associated with the metabolism of lipid and fatty acids. Notably, a lipid-droplet protein (BbLar1) maintains the intracellular homeostasis of fatty acids and is crucial to fungal tolerance to LA stress, which significantly contributes to fungal compatibility with UFAs. Additionally, BbLar1 links the lipid droplets to global expression profiles in B. bassiana under LA stress. Our investigations provide an initial framework for improving the efficacy of insect pathogenic fungi in practical application.

Biased pollen transfer by bumblebees favors the paternity of virus-infected plants in cross-pollination.

We used a green fluorescent protein marker gene for paternity analysis to determine if virus infection affected male reproductive success of tomato in bumblebee-mediated cross-pollination under glasshouse conditions. We found that bumblebees that visited flowers of infected plants showed a strong preference to subsequently visit flowers of non-infected plants. The behavior of the bumblebees to move toward non-infected plants after pollinating virus-infected plants appears to explain the paternity data, which demonstrate a statistically significant ∼10-fold bias for fertilization of non-infected plants with pollen from infected parents. Thus, in the presence of bumblebee pollinators, CMV-infected plants exhibit enhanced male reproductive success.

Evaluating the hologenome concept by analyzing the root-endosphere microbiota of chimeric plants.

The hologenome concept considers the entity formed by a host and its microbiota, the holobiont, as new level of hierarchical organization subject to neutral and selective forces. We used grafted plants to formally evaluate the hologenome concept. We analyzed the root-endosphere microbiota of two independent watermelon and grapevine plant systems, including ungrafted and reciprocal-grafting combinations. Grafted and ungrafted hosts harbor markedly different microbiota compositions. Furthermore, the results indicate a non-random assembly of bacterial communities inhabiting the root endosphere of chimeric plants with interactive effect of both the rootstock and scion on the recruitment of microorganisms. Because chimeric plants did not have a random microbiota, the null hypothesis that holobionts assemble randomly and hologenome concept is an intellectual construction only can be rejected. The study supports the relevance of hologenome as biological level of organization and opens new avenues for a better fundamental understanding of plants as holobionts.

Large-scale mRNA transfer between Haloxylon ammodendron (Chenopodiaceae) and herbaceous root holoparasite Cistanche deserticola (Orobanchaceae).

Exchanges of mRNA were shown between host and stem parasites but not root parasites. Cistanche deserticola (Orobanchaceae) is a holoparasitic herb which parasitizes on the roots of woody plant Haloxylon ammodendron (Chenopodiaceae). We used transcriptome sequencing and bioinformatic analyses to identify nearly ten thousand mobile mRNAs. Transcript abundance appears to be a driving force for transfer event and mRNA exchanges occur through haustorial junction. Mobility of selected mRNAs was confirmed in situ and in sunflower-Orobanche cumana heterologous parasitic system. Four C. deserticola →H. ammodendron mobile mRNAs appear to facilitate haustorium development. Of interest, two mobile mRNAs of putative resistance genes CdNLR1 and CdNLR2 cause root-specific hypersensitive response and retard parasite development, which might contribute to parasitic equilibrium. The present study provides evidence for the large-scale mRNA transfer event between a woody host and a root parasite, and demonstrates the functional relevance of six C. deserticola genes in host-parasite interactions.

Insect visitation and pollination of a culturally significant plant, Hopi tobacco (Nicotiana rustica).

Nicotiana rustica is a monecious member of the Solanaceae family, distributed across North America where it is grown for ceremonial purposes. Flowers of N. rustica open in the morning and are receptive to pollen until the following day. This study investigates the role of diurnal and nocturnal pollinators in reproductive success (seed weight, seed set, seediness) and insect visitation rate in N. rustica in eastern North America using floral bagging techniques. Results show that N. rustica benefits most from open (open day and night) and day open (closed at night) pollination. Fruit weight was higher with the presence of diurnal pollinators; fruit set and seediness were unaffected. Video recordings show that the most abundant floral visitors are small bees. These results provide information that will be useful in making management decisions surrounding the continued growth of this culturally significant plant species.

Root exudates enhanced rhizobacteria complexity and microbial carbon metabolism of toxic plants.

Root exudates and rhizosphere microorganisms play key roles in the colonization of toxic plants under climate change and land degradation. However, how root exudates affect the rhizosphere microorganisms and soil nutrients of toxic plants in degraded grasslands remains unknown. We compared the interaction of soil microbial communities, root exudates, microbial carbon metabolism, and environmental factors in the rhizosphere of toxic and non-toxic plants. Deterministic processes had a greater effect on toxic than non-toxic plants, as root exudates affected rhizosphere microorganisms directly. The 328 up-regulated compounds in root exudates of toxic plants affected the diversity of rhizosphere microorganisms. Rhizosphere bacteria-enriched enzymes were involved in the phenylpropanoid biosynthesis pathway. Root exudates of toxic plants form complex networks of rhizosphere microorganisms, provide high rhizosphere nutrients, and increase microbial carbon metabolism. The interaction between root exudates and rhizosphere microorganisms is the key mechanism that enables toxic plants to spread in degraded grassland habitats.

Identification of the core c-di-GMP turnover proteins responsible for root colonization of Bacillus velezensis.

Root colonization by beneficial rhizobacteria determines their plant beneficial effects. The messenger c-di-GMP is involved in the bacterial transition process between motility and biofilm, which are crucial to the colonization ability of the rhizobacteria. In this study, we identified three GGDEF domain-containing proteins (YdaK, YhcK, and YtrP) and two EAL domain-containing proteins (YuxH and YkuI) in beneficial rhizobacterium Bacillus velezensis SQR9. We found that deficiency of ytrP or ykuI in SQR9 led to impaired biofilm formation, while deficiency of yuxH led to weakened motility. Further investigation showed that YtrP, YuxH, and YkuI all contributed to the root colonization of SQR9 on cucumber root. Further bioinformatics analysis showed that YtrP and YuxH are conserved in plant beneficial Bacillus group, while they do not occur in animal pathogenic Bacillus. This research will be useful for enhancing the beneficial function of Bacillus spp. in agricultural application.