Phloem Sap Research Articles

Shoot-Silicon-Signal protein to regulate root silicon uptake in rice

Plants accumulate silicon to protect them from biotic and abiotic stresses. Especially in rice (Oryza sativa), a typical Si-accumulator, tremendous Si accumulation is indispensable for healthy growth and productivity. Here, we report a shoot-expressed signaling protein, Shoot-Silicon-Signal (SSS), an exceptional homolog of the flowering hormone “florigen” differentiated in Poaceae. SSS transcript is only detected in the shoot, whereas the SSS protein is also detected in the root and phloem sap. When Si is supplied from the root, the SSS transcript rapidly decreases, and then the SSS protein disappears. In sss mutants, root Si uptake and expression of Si transporters are decreased to a basal level regardless of the Si supply. The grain yield of the mutants is decreased to 1/3 due to insufficient Si accumulation. Thus, SSS is a key phloem-mobile protein for integrating root Si uptake and shoot Si accumulation underlying the terrestrial adaptation strategy of grasses.

Integrated Diagnostic Approach Using qPCR, ddPCR and LAMP‐Based Molecular Tools for Two Leafhopper Species Identification (Nephotettix cincticeps and Nephotettix virescens)

ABSTRACTAccurate identification of the leafhopper species Nephotettix cincticeps and N. virescens is necessary because they feed on phloem sap and transmit plant pathogens including viruses that can cause significant damage to rice plants. Initially, we examined their physical traits using a microscope and a specific camera and observed differences between the two species. To further validate these distinctions on a molecular level, we designed primers based on mitochondrial genome sequences obtained from the NCBI database. The primers were effectively used in general PCR and loop‐mediated isothermal amplification (LAMP) tests. General PCR was successfully performed at 61°C using NC_mt5117F33 and NC_mt5438B33 primers for N. cincticeps and NV_mt3089F31, NV_mt3360B31 primers for N. virescens. The effectiveness of the selected primers was validated using quantitative PCR (qPCR) on the pooled gDNA samples and droplet digital PCR (ddPCR). The best conditions for LAMP were 61°C and time 45 min, resulting in the most favourable amplification outcomes. Additionally, LAMP enables diagnosis at the individual level in the field, and qPCR and ddPCR can be used for relative or absolute quantification, even from pooled gDNA from multiple species. This study emphasises the importance of combining traditional and molecular techniques in primary pest species diagnostics to ensure precise and timely pest control, which can contribute to improved pest management strategies.

Deep Sequencing Analysis of Virome Components, Viral Gene Expression and Antiviral RNAi Responses in Myzus persicae Aphids.

The green peach aphid (Myzus persicae) is a generalist pest damaging crops and transmitting viral pathogens. Using Illumina sequencing of small (s)RNAs and poly(A)-enriched long RNAs, we analyzed aphid virome components, viral gene expression and antiviral RNA interference (RNAi) responses. Myzus persicae densovirus (family Parvoviridae), a single-stranded (ss)DNA virus persisting in the aphid population, produced 22 nucleotide sRNAs from both strands of the entire genome, including 5'- and 3'-inverted terminal repeats. These sRNAs likely represent Dicer-dependent small interfering (si)RNAs, whose double-stranded RNA precursors are produced by readthrough transcription beyond poly(A) signals of the converging leftward and rightward transcription units, mapped here with Illumina reads. Additionally, the densovirus produced 26-28 nucleotide sRNAs, comprising those enriched in 5'-terminal uridine and mostly derived from readthrough transcripts and those enriched in adenosine at position 10 from their 5'-end and mostly derived from viral mRNAs. These sRNAs likely represent PIWI-interacting RNAs generated by a ping-pong mechanism. A novel ssRNA virus, reconstructed from sRNAs and classified into the family Flaviviridae, co-persisted with the densovirus and produced 22 nucleotide siRNAs from the entire genome. Aphids fed on plants versus artificial diets exhibited distinct RNAi responses affecting densovirus transcription and flavivirus subgenomic RNA production. In aphids vectoring turnip yellows virus (family Solemoviridae), a complete virus genome was reconstituted from 21, 22 and 24 nucleotide viral siRNAs likely acquired with plant phloem sap. Collectively, deep-sequencing analysis allowed for the identification and de novo reconstruction of M. persicae virome components and uncovered RNAi mechanisms regulating viral gene expression and replication.

Genistein and Aphid Probing Behavior: Case Studies on Polyphagous Aphid Species.

(1) Background: Genistein is a naturally occurring flavonoid with a rich spectrum of biological activities, including plant-herbivore interactions. The aim of the study was to evaluate the effect of exogenous application of genistein on aphid behavior during probing in plant tissues. (2) Methods: Vicia faba, Brassica rapa ssp. pekinensis, and Avena sativa were treated transepidermally with a 0.1% ethanolic solution of genistein, and the probing behavior of generalist aphid species Aphis fabae, Myzus persicae, and Rhopalosiphum padi was monitored on their respective treated and untreated host plants using electropenetrography (=electrical penetration graph technique, EPG); (3) Results: Genistein did not deter aphid probing activities in non-phloem tissues. In A. fabae and R. padi, a trend towards reduction and in M. persicae a trend towards increase in phloem sap ingestion occurred on genistein-treated plants, but these trends were not statistically significant. (4) Conclusions: Genistein is not a deterrent chemical against generalist aphid species studied; therefore, it is not recommended for practical application.

Evaluation of Different Mode of Action Insecticides for the Control of Bemisia tabaci; Enhancement of Pesticide Efficacy.

Bemisia tabaci (Gennadius) is a major pest worldwide, causing damage to a vast range of plants through its feeding on phloem sap and its vectoring of >100 plant viruses. Although not established in the UK, it is regularly introduced on planting material, which poses a significant plant health risk. Restrictions on pesticide use and increasing resistance to available active ingredients limit options for effective control of potential outbreaks. Alternative management options are required to mitigate this risk. There was high variability in the efficacy of the different modes of action products tested against two life stages (adults and larvae) as well as the Middle East-Asia Minor 1 (MEAM1) and Mediterranean (MED) cryptic species of B. tabaci. For both adults and larvae, MEAM1 were more susceptible than MED insects, possibly due to differences in resistance developed against some active ingredients. All products tested were effective to varying degrees against MEAM1 adults with Tracer (spinosad), PREV-AM (orange oil), Sequoia (sulfoxaflor), and FLiPPER (fatty acids) having similar efficacies (59-78% mortality). In contrast, PREV-AM and FLiPPER were most effective against MED adults (74% and 65% mortalities, respectively). Both MED and MEAM1 larvae were highly susceptible to FLiPPER and PREV-AM (>95% mortality), and the efficacy of Tracer and FLiPPER can be enhanced by using in combination with PREV-AM, and this can be achieved by using low doses of each product. Synergy was measured between PREV-AM and Tracer against MEAM1 larvae, which has the potential to provide effective control with a reduced pesticide application.

Virus-Induced Gene Silencing Simultaneously Exploits ‘Attract and Kill’ Traits in Plants and Insects to Manage Huanglongbing

Abstract The vector-borne disease Huanglongbing (HLB) causes severe economic losses to citrus production worldwide with no available cure. Herein, we applied virus-induced gene silencing technology to engineer citrus that preferentially attracted and specifically killed Diaphorina citri, the vector associated with HLB. We engineered the infectious citrus tristeza virus (CTV-T36) clone to carry three truncated genes. The triple construct (CTV-tAwd-tWnt-tPDS) produces small interfering RNAs (siRNAs) against phytoene desaturase, PDS, to yield a phenotype with visual, olfactory, and gustatory cues that preferentially attracted D. citri. In addition, siRNAs targeted two genes related to flight in D. citri, abnormal wing disc (DcAwd) and wingless (DcWnt), that caused wing malformations and decreased survival in psyllids that fed on plants inoculated with the engineered virus. During two successive generations, D. citri reared on CTV-tAwd-tWnt-tPDS-inoculated plants exhibited higher mortality across life stages as well as reduced fecundity and fertility as compared with those reared on non-infected plants or CTV-wt-inoculated plants. Furthermore, CTV-tAwd-tWnt-tPDS-inoculated plants shortened the lifespan of D. citri by more than 20 days. Morphological abnormalities were noted in those adults that did successfully emerge on plants inoculated with CTV-tAwd-tWnt-tPDS, including cocked wings with a bowl-shaped depression and/or a convex shape. Phloem sap from CTV-tAwd-tWnt-tPDS-inoculated plants decreased the survival of D. citri adults, confirming that siRNAs were present in the sap of these plants. Collectively, we provide proof of concept for a novel variant of the attract-and-kill method where the cultivated crop is potentially transformed into a hyper-attractive population and transmission sink for a phytopathogen vector.

Influence of Previous Infestation of Wheat Leaves and Ears by Sitobion avenae on Interaction with Rhopalosiphum padi.

Different herbivorous species that share a host plant may interact via competition or facilitation, depending on whether the interaction partners are hindered by or benefit from the interaction. Sap-sucking insects, such as aphids, can influence each other indirectly by altering the composition of the shared phloem sap. Aphid-induced changes in the plant may affect aphid performance and lead to a shift in the balance between different co-occurring aphid species. In this study, we compared the performance of the English grain aphid (Sitobion avenae) and the bird cherry-oat aphid (Rhopalosiphum padi) simultaneously infesting leaves or ears of wheat (Triticum aestivum) plants, which had been either previously infested by S. avenae or kept uninfested. Colonies of S. avenae were larger on ears than on leaves, while the opposite pattern was found for R. padi. Pre-infestation of ears, but not of leaves, by S. avenae led to a higher total aphid number and colony size of S. avenae at some time points. The balance between the two species was only slightly affected by previous infestation at some time points. The findings of this study contribute to the understanding of plant-aphid as well as aphid-aphid interactions in agricultural fields.

Interplay between a polerovirus and a closterovirus decreases aphid transmission of the polerovirus.

Multi-infection of plants by viruses is very common and can change drastically infection parameters such as virus accumulation, distribution, and vector transmission. Sugar beet is an important crop that is frequently co-infected by the polerovirus beet chlorosis virus (BChV) and the closterovirus beet yellows virus (BYV), both vectored by the green peach aphid (Myzus persicae). These phloem-limited viruses are acquired while aphids ingest phloem sap from infected plants. Here we found that co-infection decreased transmission of BChV by ~50% but had no impact on BYV transmission. The drastic reduction of BChV transmission was due to neither lower accumulation of BChV in co-infected plants nor reduced phloem sap ingestion by aphids from these plants. Using the signal amplification by exchange reaction fluorescent in situ hybridization technique on plants, we observed that 40% of the infected phloem cells were co-infected and that co-infection caused redistribution of BYV in these cells. The BYV accumulation pattern changed from distinct intracellular spherical inclusions in mono-infected cells to a diffuse form in co-infected cells. There, BYV co-localized with BChV throughout the cytoplasm, indicative of virus-virus interactions. We propose that BYV-BChV interactions could restrict BChV access to the sieve tubes and reduce its accessibility for aphids and present a model of how co-infection could alter BChV intracellular movement and/or phloem loading and reduce BChV transmission.IMPORTANCEMixed viral infections in plants are understudied yet can have significant influences on disease dynamics and virus transmission. We investigated how co-infection with two unrelated viruses, BChV and BYV, affects aphid transmission of the viruses in sugar beet plants. We show that co-infection reduced BChV transmission by about 50% without affecting BYV transmission, despite similar virus accumulation rates in co-infected and mono-infected plants. Follow-up experiments examined the localization and intracellular distribution of the viruses, leading to the discovery that co-infection caused a redistribution of BYV in the phloem vessels and altered its repartition pattern within plant cells, suggesting virus-virus interactions. In conclusion, the interplay between BChV and BYV affects the transmission of BChV but not BYV, possibly through direct or indirect virus-virus interactions at the cellular level. Understanding these interactions could be crucial for managing virus propagation in crops and preventing yield losses.

GABA-Decorated Nanocarrier for Smart Delivery of Fludioxonil for Targeted Control of Banana Wilt Disease.

Developing a targeted nanopesticide to control the vascular disease of banana in agriculture is crucial to improve pesticide utilization. In this study, according to the degree of functionalization, three γ-aminobutyric acid (GABA)-decorated nanocarriers (PSI-GABA8, PSI-GABA18, and PSI-GABA28) were constructed for smart delivery of nonsystemic fungicide in banana phloem tissues. Fludioxonil (Flu) was loaded in nanocarriers to form Flu@PSI-GABA nanoparticles with a core/shell structure for control of banana wilt disease. Results demonstrated that the delivery dosage of Flu was up to 1.6 mg/L in castor phloem sap using PSI-GABA28 nanocarriers. In vitro results showed that the EC50 of Flu@PSI-GABA28 was 0.0116 mg/L, and the inhibitory activity was about 8.8 times higher than that of technical-grade (TC) Flu. Flu@PSI-GABA28 could be transported for long distances and accumulated to the rhizome of banana by foliar application, and the control effectiveness was about 20 times that of the conventional Flu (50% WP) for the banana wilt. This study provides a distinctive guidance for effective control of vascular diseases in precision agriculture application.

Phloem sap from melon plants contains extracellular vesicles that carry active proteasomes which increase in response to aphid infestation.

The morphogenesis of higher plants requires communication among distant organs throughout vascular tissues (xylem and phloem). Numerous investigations have demonstrated that phloem also act as a distribution route for signalling molecules being observed that different macromolecules translocated by the sap, including nucleic acids and proteins, change under stress situations. The participation of extracellular vesicles (EVs) in this communication has been suggested, although little is known about their role. In fact, in the last decade, the presence of EVs in plants has originated a great controversy, where major concerns arose from their origin, isolation methods, and even the appropriate nomenclature for plant nanovesicles. Phloem sap exudates from melon plants, either aphid-free or infested with Aphis gossypii, were collected by stem incision. After sap concentration (Amicon), phloem EVs (PhlEVs) were isolated by size exclusion chromatography. PhlEVs were characterised using Nanoparticle Tracking Analysis, Transmission electron microscopy and proteomic analysis. Here we confirm the presence of EVs in phloem sap in vivo and the detection of changes in the particles/protein ratio and composition of PhlEVs in response to insect feeding, revealing the presence of typical defence proteins in their cargo as well as components of the proteasome complex. PhlEVs from infested plants showed lower particles/protein ratio and almost two times more proteolytic activity than PhlEVs from aphid-free plants. In both cases, such activity was inhibited in a dose-dependent manner by the proteasome inhibitor MG132. Our results suggest that plants may use this mechanism to prepare themselves to receive infectious agents and open up the possibility of an evolutionary conserved mechanism of defence against pathogens/stresses in eukaryotic organisms.

Identification and function of microRNAs in hemipteran pests: A review.

Hemiptera is one of the most significant orders of insect pests, including whiteflies, true bugs, aphids, planthoppers, psyllids, and so forth, which have led to substantial economic losses in agricultural industries and have significantly affected food yields through their ability to suck the phloem sap of crops and transmit numerous bacterial and viral pathogens. Therefore, explorations of pest-specific, eco-friendly and easy-to-adopt technologies for hemipteran pest control are urgently needed. To the best of our knowledge, microRNAs (miRNAs), which are endogenous non-coding small RNAs approximately 22 nucleotides in length, are involved in regulating gene expression via the direct recognition and binding of the 3'-untranslated region (3'-UTR) of target messenger RNAs (mRNAs) or by acting as a center of a competitive endogenous RNA (ceRNA) network at the post-transcriptional level. This review systematically outlines the characterization and functional investigation of the miRNA biogenesis pathway in hemipteran pests, such as whiteflies, true bugs, aphids and planthoppers. In addition, we explored the results of small RNA sequencing and functional observations of miRNAs in these pests, and the results suggest that the numerous miRNAs obtained and annotated via high-throughput sequencing technology and bioinformatic analyses contribute to molting development, fitness, wing polyphenism, symbiont interactions and insecticide resistance in hemipteran pests. Finally, we summarize current advances and propose a framework for future research to extend the current data and address potential limitations in the investigation and application of hemipteran miRNAs.

A Transmission Assay of 'Candidatus Liberibacter asiaticus' Using Citrus Phloem Sap and Topical Feeding to Its Insect Vector, Diaphorina citri.

'Candidatus Liberibacter asiaticus', the putative causal agent of citrus greening disease, is transmitted by the Asian citrus psyllid, Diaphorina citri, in a propagative, circulative, and persistent manner. Unfortunately, 'Ca. L. asiaticus' is not yet available in pure culture to carry out Koch's postulates and to confirm its etiology. When a pure culture is available, an assay to test its infectivity in both the insect vector and the plant host will be crucial. Herein, we described a transmission assay based on the use of phloem sap extracted from infected citrus plants and topical feeding to D. citri nymphs. Phloem sap was collected by centrifugation, diluted with 0.1 M phosphate buffer pH 7.4 containing 20% (wt/vol) sucrose and 0.1% ascorbic acid (wt/vol) as an antioxidant, and delivered to third through fifth instar nymphs by placing droplets on the mouthparts. Nymphs unfolded the stylets and acquired the phloem sap containing the bacterial pathogen. Nymphs were then placed onto Citrus macrophylla seedlings (10 nymphs per seedling) for an inoculation period of 2 weeks. A transmission rate of up to 80% was recorded at 6 months postinoculation. The method could be a powerful tool to test the transmissibility of the bacterial pathogen after various treatments to reduce the viability of the bacteria or to block its transmission. In addition, it might be a potent assay to achieve Koch's postulates if a pure culture of 'Ca. L. asiaticus' becomes available.

Insecticide-free reduction of Myzus persicae aphids on sweet pepper (Capsicum annuum L.) through MgSO4-faciliated amino acid and sugar reallocation

Fruits of sweet pepper (Capsicum annuum L.) are among the most consumed fresh fruits, however, their production can be affected by aphids, which feed on the amino acids in the leaves. Mg2+ in leaves plays a crucial role in the reallocation of amino acids from source organs (leaves) to sink organs (fruits). We hypothesize that foliar application of Mg2+ can reduce free amino acid concentration in leaves by promoting their transfer to sink fruits, thereby compromising aphid colonization by reducing their nutritional basis. To test this, a two-factorial experiment was conducted with sweet pepper subjected to aphid infestation with and without leaf MgSO4 application. The concentration of amino acids, sugars, organic acids and mineral nutrients was analysed in the leaves, leaf phloem sap and fruits. Mg2+application decreased the concentrations of amino acids and sugars that are key for aphid nutrition in both the leaf and phloem sap and increased the size and number fruits, indicating an increased sink strength in response to Mg2+. This response was accompanied by a reduced number of aphids on the entire plant. In results, the sugar and amino acid concentration in the phloem got less attractive for the aphids. We conclude that Mg2+ worsens the nutritional situation for aphids by facilitating the reallocation of metabolites to form fruits.

Niche construction and niche choice by aphids infesting wheat ears

The niche of aphids is largely defined by their consumption of plant phloem sap and its composition, including nutrients and specialized metabolites. Niche construction is the change of the environment by organisms, which may influence the fitness of these organisms and their offspring. To better understand interactions between plants and aphids, it is necessary to investigate whether aphids modify the chemical composition of the phloem sap of their host plants and whether conspecifics are affected by previous infestation. In the current study, ears of wheat (Triticum aestivum) plants were infested with clonal lineages of the English grain aphid (Sitobion avenae) or were left uninfested. The metabolic composition of ear phloem sap exudates was analyzed through amino acid profiling and metabolic fingerprinting. Aphids of the clonal lineages were either put on previously aphid-infested or on uninfested ears and their colony sizes followed over time. Furthermore, it was investigated whether aphids choose one treatment group over another. Sitobion avenae infestation affected the relative concentrations of some metabolites in the phloem exudates of the ears. Compared to uninfested plants, the relative concentration of asparagine was higher after aphid infestation. Colonies grew significantly larger on previously aphid-infested ears, which the aphids also clearly chose in the choice experiment. The pronounced positive effect of previous infestation on aphid colonies indicates niche construction, while the choice of these constructed niches reveals niche choice by S. avenae on wheat. The interplay between these different niche realization processes highlights the complexity of interactions between aphids and their hosts.

Identification and characterization of transition metal-binding proteins and metabolites in the phloem sap of Brassica napus

Transition metal (TM) distribution through the phloem is an essential part of plant metabolism and is required for systemic signaling and balancing source-to-sink relationships. Due to their reactivity, TMs are expected to occur in complexes within the phloem sap; however, metal speciation in the phloem sap remains largely unexplored. Here, we isolated phloem sap from Brassica napus and analyzed it via size exclusion chromatography (SEC) coupled online to sector-field ICP-MS. Our data identified known TM binding proteins and molecules including metallothioneins (MT), glutathione, and nicotianamine. While the main peak of all metals was low MW (∼1.5 kD), additional peaks ∼10-15 kD containing Cu, Fe, S and Zn were also found. Further physicochemical analyses of MTs with and without affinity tags corroborated that MTs can form complexes of diverse molecular weights. We also identified and characterized potential artifacts in the TM-biding ability of B. napus MTs between tagged and non-tagged MTs. That is, the native BnMT2 binds Zn, Cu and Fe, while MT3a and MT3b only bind Cu and Zn. In contrast, his-tagged MTs bind less Cu and were found to bind Co and Mn and aggregated to oligomeric forms to a greater extent compared to the phloem sap. Our data indicates that TM chemistry in the phloem sap is more complex than previously anticipated and that more systematic analyses are needed to establish the precise speciation of TM and TM-ligand complexes within the phloem sap.

Antioxidant Enzyme, Transcriptomic, and Metabolomic Changes in Lily (Lilium spp.) Leaves Induced by Aphis gossypii Glover.

Cotton aphids (Aphis gossypii Glover) cause harm by feeding on phloem sap and spreading plant viruses to lily. Understanding the mechanisms by which aphids infest lily plants is crucial for effective aphid management and control. In this study, we investigated the activity of antioxidants, integrated nontargeted metabolomes and transcriptomes of lilies infested by cotton aphids to explore the changes in lily leaves. Overall, the results indicated that the catalase (CAT) activity in the leaves of the lily plants was greater than that in the leaves of the control plants. A comprehensive identification of 604 substances was conducted in the leaves. Furthermore, the differentially abundant metabolite analysis revealed the enrichment of phenylalanine metabolism and α-linolenic acid metabolism. Moreover, 3574 differentially expressed genes (DEGs), whose expression tended to increase, were linked to glutathione metabolism and phenylpropanoid biosynthesis. In addition, the integrated analysis revealed that the defensive response of lily leaves to aphids is manifested through antioxidant reactions, phenylpropane and flavonoid biosynthesis, and α-linolenic acid metabolism. Finally, the key metabolites were CAT, glutathione, coumaric acid, and jasmonic acid, along with the key genes chalcone synthase (CHS), phenylalanine ammonia-lyase (PAL), and 12-oxo-phytodienoic acid reductase (OPR). Accordingly, the findings of this research elucidate the molecular and metabolic reactions of A. gossypii in lily plants, offering valuable insights for developing aphid resistance strategies in lily farming.

Dynamic response of essential amino acid biosynthesis in Buchnera aphidicola to supplement sub-optimal host nutrition

The endosymbiotic bacterium Buchnera aphidicola allows its host Acyrthosiphon pisum to utilise a nutritionally limited phloem sap diet without significant mortality by providing essential amino acids (EAAs), which it biosynthesises de novo via complex pathways consisting of multiple enzymes. Previous studies have reported how non-essential amino acids (NEAAs) provided by the host are utilised by B. aphidicola, along with how genes within the biosynthetic pathways respond to amino acid deficiency. Although the effect on B. aphidicola gene expression upon the removal of a single EAA and multiple NEAAs from the A. pisum diet has been reported, little is known about the effects of the complete simultaneous removal of multiple EAAs, especially branched-chain amino acids (BCAAs). To investigate this, A. pisum was provided with amino acid deficient diets ilv- (lacking isoleucine, leucine, valine) or thra- (lacking threonine, methionine, lysine). Due to their involvement in the production of several amino acids, the expression of genes ilvC, ilvD (both involved in isoleucine, leucine and valine biosynthesis) and thrA (involved in threonine, methionine and lysine biosynthesis) was analysed and the expression of trpC (involved in tryptophan biosynthesis) was used as a control. Survival was reduced significantly when A. pisum was reared on ilv- or thra- (P < 0.001 and P = 0.000 respectively) compared to optimal artificial diet and was significantly lower on ilv- (P < 0.001) than thra-. This is likely attributed to the EAAs absent from ilv- being required at higher concentrations for aphid growth, than those EAAs absent from thra-. Expression of ilvC and ilvD were upregulated 2.49- and 2.08-fold (respectively) and thrA expression increased 2.35- and 2.12-fold when A. pisum was reared on ilv- and thra- (respectively). The surprisingly large upregulation of thrA when reared on ilv- is likely due to threonine being an intermediate in isoleucine biosynthesis. Expression of trpC was not affected by rearing on either of the two amino acid deficient diets. To our knowledge this study has shown, for the first time, how genes within the biosynthetic pathways of an endosymbiont respond to the simultaneous complete omission of multiple EAAs as well as all three BCAAs (leucine, isoleucine, valine), from the host diet.

Co-mutation of OsLPR1/3/4/5 provides a promising strategy to minimize Cd contamination in rice grains

Minimizing cadmium (Cd) contamination in rice grains is crucial for ensuring food security and promoting sustainable agriculture. Utilizing genetic modification to generate rice varieties with low Cd accumulation is a promising strategy due to its cost-effectiveness and operational simplicity. Our study demonstrated that the CRISPR-Cas9-mediated quadruple mutation of the multicopper oxidase genes OsLPR1/3/4/5 in the japonica rice cultivar Tongjing 981 had little effect on yields. However, a notable increase was observed in the cell wall functional groups that bind with Cd. As a result, the quadruple mutation of OsLPR1/3/4/5 enhanced Cd sequestration within the cell wall while reducing Cd concentrations in both xylem and phloem sap, thereby inhibiting Cd transport from roots to shoots. Consequently, Cd concentrations in brown rice and husk in oslpr1/3/4/5 quadruple mutants (qm) decreased by 52% and 55%, respectively, compared to the wild-type. These findings illustrate that the quadruple mutation of OsLPR1/3/4/5 is an effective method for minimizing Cd contamination in rice grains without compromising yields. Therefore, the quadruple mutation of OsLPR1/3/4/5 via biotechnological pathways may represent a valuable strategy for the generation of new rice varieties with low Cd accumulation.

Genome-Wide Identification and Expression Pattern of Sugar Transporter Genes in the Brown Planthopper, Nilaparvata lugens (Stål).

Sugar transporters play important roles in controlling carbohydrate transport and are responsible for mediating the movement of sugars into cells in numerous organisms. In insects, sugar transporters not only play a role in sugar transport but may also act as receptors for virus entry and the accumulation of plant defense compounds. The brown planthopper, Nilaparvata lugens, inflicts damage on rice plants by feeding on their phloem sap, which is rich in sugars. In the present study, we identified 34 sugar transporters in N. lugens, which were classified into three subfamilies based on phylogenetic analysis. The motif numbers varied from seven to eleven, and motifs 2, 3, and 4 were identified in the functional domains of all 34 NlST proteins. Chromosome 1 was found to possess the highest number of NlST genes, harboring 15. The gut, salivary glands, fat body, and ovary were the different tissues enriched with NlST gene expression. The expression levels of NlST2, 3, 4, 7, 20, 27, 28, and 31 were higher in the gut than in the other tissues. When expressed in a Saccharomyces cerevisiae hexose transporter deletion mutant (strain EBY.VW4000), only ApST4 (previously characterized) and NlST4, 28, and 31 were found to transport glucose and fructose, resulting in functional rescue of the yeast mutant. These results provide valuable data for further studies on sugar transporters in N. lugens and lay a foundation for finding potential targets to control N. lugens.

Population dynamics, feeding strategies, and coccinellid predators of the larch woolly adelgid exules on the European larch

The larch woolly adelgid Adelges laricis Vallot (Hemiptera: Adelgidae) infests spruce Picea spp. and larch Larix spp. in the Northern Hemisphere. The present field and laboratory research embraced the hitherto unknown aspects of the biology and ecology of A. laricis on its secondary host, the European larch Larix decidua Mill. in Poland. The study showed that the A. laricis population on L. decidua was represented by exulis sistens females at dwarf stem bases, exulis progrediens mobile 1st instar nymphs (crawlers) on needles, exulis progrediens sessile “woolly” nymphs and “woolly” adults on needles, and winged sexupara females. The maximum abundance of the A. laricis population occurred from the end of April until mid-May, which coincided with the predomination of crawlers in the age structure of the population. The most important limiting weather component was high rainfall, affecting mainly the crawlers. Eight species of Coccinellidae were recorded on adelgid-infested larch trees and Exochomus quadripustulatus (L.) was the dominant species. The main differences in food consumption strategies among adelgid developmental stages were related to the duration of phloem sap ingestion. While the total duration of sap consumption at a given time was similar in all adelgid instars studied, the individual bouts of sap ingestion were much shorter in crawlers than in the sessile forms. The specific probing behavior of the mobile nymphs probably reflects the crawlers’ strategy to evaluate plant sap quality in different phloem vessels. It is likely that such strategy is a behavioral preadaptation for finding abundant food sources for permanent settling before crawlers develop into sessile nymphs and adult females. The feeding strategy of crawlers promotes the rapid dispersal of the population. Therefore, the control of A. laricis on L. decidua should focus on the early stages of the population development, which would most effectively reduce the final damage.