Identification, expression analysis, and functional verification of three opsin genes related to the phototactic behaviour of Ostrinia furnacalis.

Ostrinia furnacalis is a disreputable herbivorous pest that poses a serious threat to corn crops. Phototaxis in nocturnal moths plays a crucial role in pest prediction and control. Insect opsins are the main component of insect visual system. However, the inherent molecular relationship between phototactic behaviour and vision of insects remains a mystery. Herein, three opsin genes were identified and cloned from O. furnacalis (OfLW, OfBL, and OfUV). Bioinformatics analysis revealed that all opsin genes had visual pigment (opsin) retinal binding sites and seven transmembrane domains. Opsin genes were distributed across different developmental stages and tissues, with the highest expression in adults and compound eyes. The photoperiod-induced assay elucidated that the expression of three opsin genes in females were higher during daytime, while their expression in males tended to increase at night. Under the sustained darkness, the expression of opsin genes increased circularly, although the increasing amplitude in males was lower when compared with females. Furthermore, the expression of OfLW, OfBL, and OfUV was upregulated under green, blue, and ultraviolet light, respectively. The results of RNA interference showed that the knockout of opsin genes decreased the phototaxis efficiency of female and male moths to green, blue, and ultraviolet light. Our results reveal that opsin genes are involved in the phototactic behaviour of moths, providing a potential target gene for pest control and a basis for further investigation on the phototactic behaviour of Lepidoptera insects.

Terrihabitans rhizophilus sp. nov., isolated from the rhizosphere soil of plant in temperate semi-arid steppe.

A bacterial strain PJ23T was isolated from the rhizosphere soil of Elymus dahuricus Turcz. sampled from a temperate semi-arid steppe in the northern of Inner Mongolia Autonomous Region, China. The strain is Gram-stain-negative, aerobic, light-pink, short rod-shaped, and non-spore-forming. Cell growth could be observed at 4-29℃ (optimal at 24℃), pH 6.0-8.6 (optimal at 8.0) and in the presence of 0-5.0% (w/v) NaCl (optimal at 2.5%). The major cellular fatty acids of strain PJ23T were Summed feature 8 (C18:1 ω6c and/or C18:1 ω7c) (39.42%) and C16:0 (9.60%). The polar lipids were phosphatidylcholine, two unidentified glycolipids, one unidentified aminophospholipid, and two other unidentified polar lipids. The major respiratory quinone was ubiquinone-10. Phylogeny analysis based on 16S rRNA gene sequences retrieved from the genomes showed that, the strain was closely related to the species Terrihabitans soli IZ6T and Flaviflagellibacter deserti SYSU D60017T, with the sequence similarities of 96.79% and 96.15%, respectively. The G + C content was 65.23 mol% calculated on draft genome sequencing. Between the strains PJ23T and Terrihabitans soli IZ6T, the average nucleotide identity (ANI), amino acid identity (AAI) and digital DNA-DNA hybridization (dDDH) was 73.39%,71.12% and 15.7%, these values were lower than the proposed and generally accepted species boundaries of ANI, AAI and dDDH, respectively. Based on phenotypic, chemotaxonomic, and phylogenetic characteristics, strain PJ23T represents a novel species of Terrihabitans, for which the name Terrihabitans rhizophilus sp. nov. is proposed. The type strain is PJ23T (= KCTC 92977 T = CGMCC 1.61577 T).

Identification of Six Small Heat Shock Protein Genes in Ostrinia furnacalis (Lepidoptera: Pyralidae) and Analysis of Their Expression Patterns in Response to Environmental Stressors.

Ostrinia furnacalis (Guenée) is a major insect pest in maize production that is highly adaptable to the environment. Small heat shock proteins (sHsps) are a class of chaperone proteins that play an important role in insect responses to various environmental stresses. The present study aimed to clarify the responses of six O. furnacalis sHsps to environmental stressors. In particular, we cloned six sHsp genes, namely, OfHsp24.2, OfHsp21.3, OfHsp20.7, OfHsp21.8, OfHsp29.7, and OfHsp19.9, from O. furnacalis. The putative proteins encoded by these genes contained a typical α-crystallin domain. Real-time quantitative polymerase chain reaction was used to analyze the differences in the expression of these genes at different developmental stages, in different tissues of male and female adults, and in O. furnacalis under UV-A and extreme temperature stresses. The six OfsHsp genes were expressed at significantly different levels based on the developmental stage and tissue type in male and female adults. Furthermore, all OfsHsp genes were significantly upregulated in both male and female adults under extreme temperature and UV-A stresses. Thus, O. furnacalis OfsHsp genes play important and unique regulatory roles in the developmental stages of the insect and in response to various environmental stressors.

Expression stability of candidate RT-qPCR housekeeping genes in Spodoptera frugiperda (Lepidoptera: Noctuidae).

Reverse-transcription quantitative polymerase chain reaction (RT-qPCR) is commonly used to quantify gene expression. For normalization, the expression of each gene is compared with a reference "housekeeping" gene that is stably expressed under relevant stress. Unfortunately, there have been no reports on the stability of such reference genes under various treatments of the Spodoptera frugiperda. In this study, we used five tools (RefFinder, GeNorm, NormFinder, BestKeeper, and ΔCt methods) to evaluate the stability of 12 candidate reference genes (RPS18, ß-tubulin, GAPDH, RPS7, RPS15, RPL7, RPL32, Actin-5C, EF1-α, EF1-γ, RPL27, and ACE) in different instars, tissues, and treatments (high and low temperature, UV-A, and emamectin benzoate). Several ribosomal proteins (RPS7, RPS15, RPL32, RPS18, and RPL7), GAPDH, Actin-5C, and ß-tubulin, were relatively stable, suggesting that they are ideal housekeeping genes for various treatments. ACE was extremely unstable under various experimental treatments, rendering it unsuitable as an internal reference. This study identified the reference housekeeping genes stably expressed by S. frugiperda under different treatments, thus setting a foundation for further exploration of the physiological and biochemical mechanisms.

Transcriptome Analysis of Myzus persicae to UV-B Stress.

As an environmental stress factor, ultraviolet-B (UV-B) radiation directly affects the growth and development of Myzus persicae (Sulzer) (Homoptera: Aphididae). How M. persicae responds to UV-B stress and the molecular mechanisms underlying this adaptation remain unknown. Here, we analyzed transcriptome data for M. persicae following exposure to UV-B radiation for 30 min. We identified 758 significant differentially expressed genes (DEGs) following exposure to UV-B stress, including 423 upregulated and 335 downregulated genes. In addition, enrichment analysis using the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes databases illustrated that these DEGs are associated with antioxidation and detoxification, metabolic and protein turnover, immune response, and stress signal transduction. Simultaneously, these DEGs are closely related to the adaptability to UV-B stress. Our research can raise awareness of the mechanisms of insect responses to UV-B stress.

Aerophototrophica crusticola gen. nov., sp. nov., isolated from desert biocrusts.

A pink-pigmented, Gram-stain-negative, rod-shaped, strictly aerobic bacterial strain MIMtkB3T, was isolated from moss crusts in Hunshandake desert of China. Cells grew at 15-45 °C (optimum of 28 °C), at pH of 6.0-8.5 (optimum of 7.0) and with 0-1.0 % (w/v) NaCl (optimum of 0 %). The strain could biosynthesize the green-coloured pigment bacteriochlorophyll a (BChl a). The respiratory quinone was ubiquinone Q-10, while C18 : 1 ω7c and C18 : 1 2OH were the major fatty acids. Phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, an unidentified aminophospholipid, one unidentified phospholipid, three unidentified glycolipid and one unidentified lipid were the major polar lipids. Strain MIMtkB3T was most closely related to Oleisolibacter albus NAU-10T, Niveispirillum fermenti CC-LY736T, and Rhodocista centenaria SW of the family Rhodospirillaceae with 16S rRNA gene similarities of 93.09, 92.02 and 91.73%, respectively. The genomic DNA G+C content calculated on complete genome sequencing was 69.3 mol%. The average nucleotide identity between strain MIMtkB3T and its closely related type strains in Rhodospirillaceae was below 77.96 % and digital DNA-DNA hybridization lower than 24.70 %. Full light utilization pathway of aerobic anoxygenic phototrophic bacteria was identified in the genome. Based on phenotypic, chemotaxonomic and phylogenetic characteristics, strain MIMtkB3T represents a novel genus of the family Rhodospirillaceae, for which the name Aerophototrophica crusticola gen. nov., sp. nov. is proposed. The type strain is MIMtkB3T (=KCTC 42633T=MCCC 1K00570T).

Comparative Transcriptome Profiling of Resistant and Susceptible Sugarcane Cultivars in Response to Infection by Xanthomonas albilineans.

Sugarcane (Saccharum spp. hybrids) is a major source of sugar and renewable bioenergy crop worldwide and suffers serious yield losses due to many pathogen infections. Leaf scald caused by Xanthomonas albilineans is a major bacterial disease of sugarcane in most sugarcane-planting countries. The molecular mechanisms of resistance to leaf scald in this plant are, however, still unclear. We performed a comparative transcriptome analysis between resistant (LCP 85-384) and susceptible (ROC20) sugarcane cultivars infected by X. albilineans using the RNA-seq platform. 24 cDNA libraries were generated with RNA isolated at four time points (0, 24, 48, and 72 h post inoculation) from the two cultivars with three biological replicates. A total of 105,783 differentially expressed genes (DEGs) were identified in both cultivars and the most upregulated and downregulated DEGs were annotated for the processes of the metabolic and single-organism categories, respectively. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of the 7612 DEGs showed that plant-pathogen interaction, spliceosome, glutathione metabolism, protein processing in endoplasmic reticulum, and plant hormone signal transduction contributed to sugarcane's response to X. albilineans infection. Subsequently, relative expression levels of ten DEGs determined by quantitative reverse transcription-PCR (qRT-PCR), in addition to RNA-Seq data, indicated that different plant hormone (auxin and ethylene) signal transduction pathways play essential roles in sugarcane infected by X. albilineans. In conclusion, our results provide, for the first time, valuable information regarding the transcriptome changes in sugarcane in response to infection by X. albilineans, which contribute to the understanding of the molecular mechanisms underlying the interactions between sugarcane and this pathogen and provide important clues for further characterization of leaf scald resistance in sugarcane.

Sphingomonas crusticola sp. nov., isolated from biological soil crusts.

A yellow-pigmented, Gram-stain-negative, short-rod-shaped bacterial strain, MIMD3T, was isolated from biological soil crusts collected in Liangcheng, north-western China. Cell growth could be observed at 10-37 °C (optimum 25 °C), at pH 5-8 (optimum 6.6) and in the presence of 1 % (w/v) NaCl (optimum 0 %). The genomic DNA G+C content was 65.0 mol%. Analysis of 16S rRNA gene sequences showed that strain MIMD3T shared the highest similarity with Sphingomonas vulcanisoli KCTC 42454T (95.1 %), Sphingomonas oligophenolica JCM 12082T (94.8 %), Sphingomonas mali IFO 15500T (94.5 %), Sphingomonas. leidyi ATCC 15260T (94.4 %) and Sphingomonas formosensis CC-Nfb-2T (94.3 %). The strain had Q-10 as the predominant respiratory quinone, and sym-homospermidine as the major polyamine. The major fatty acids of the strain were summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c), C19 : 0 cyclo ω8c, C14 : 0 2-OH and C16 : 0. The main polar lipids of strain MIMD3T were phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine and sphingoglycolipid. Based on phenotypic, chemotaxonomic and phylogenetic characteristics, it is concluded that strain MIMD3T represents a novel species of the genus Sphingomonas, for which the name Sphingomonas crusticola sp. nov. is proposed. The type strain is MIMD3T (=KCTC 42801T=MCCC 1K01310T).

Larkinella soli sp. nov., isolated from biological soil crusts.

A novel bacterial strain, designated MIMbqt9T, was isolated from biological soil crusts collected in Erdos Plateau in Inner Mongolia. Cells were light pink, rod-shaped, aerobic, Gram-reaction-negative, motile by gliding, oxidase- and catalase-positive. Cell growth could be observed at 5-40 °C (optimum at 30 °C), at pH 4.0-6.0 (optimum at 6.0) and with 1 % (w/v) NaCl. Phylogenetic analysis of 16S rRNA gene sequence showed that strain MIMbqt9T was most closely related to the members of genus Larkinella, including L. bovis M2T2B15T (92.06 %), L. arboricola Z0532T (92.00 %) and L. insperata LMG 22510T (91.94 %). The major cellular fatty acids of strain MIMbqt9T were C16 : 1ω5c, iso-C15 : 0 and iso-C17 : 0 3-OH. Polar lipids of the strain MIMbqt9T were mainly composed of phosphatidylethanolamine (PE), phosphatidylserine (PS), two unidentified aminolipids (AL1 and AL2) and two unidentified phospholipids (PL1 and PL2). Menaquinone 7 (MK-7) was the major respiratory quinone. The G+C content of the chromosomal DNA was 57.63 mol%. On the basis of morphology, physiology, fatty acid composition, phylogeny and 16S rRNA gene sequence analysis, the isolates are assigned to a novel species of the genus Larkinella, for which the name Larkinella soli sp. nov. is proposed. The type strain is MIMbqt9T (=MCCC 1K01309T=KCTC 42800T).

Differential protein expression in the susceptible and resistant Myzus persicae (Sulzer) to imidacloprid.

Myzus persicae, a serious economic agricultural pest, has developed resistance to imidacloprid (IMI), which was widely used to control this aphid worldwide. To gain a better understanding of the mechanisms of IMI resistance in M. persicae, we carried out a comparative proteomic analysis. Total proteins of the IMI-susceptible and resistant strains were extracted and separated by two-dimensional gel electrophoresis. More than 1300 protein spots were reproducibly detected, including 14 that were more abundant and 14 less abundant. Mass spectrometry analysis and database searching helped us to identify 25 differentially abundant proteins. The identified proteins were categorized into several functional groups including signal transduction, RNA processing, protein processing, transport processing, stress response, metabolisms, and cytoskeleton structure, etc. This study is the first analysis of differentially expressed proteins in IMI-susceptible and resistant M. Persicae, and gives new insights into the mechanisms of IMI resistance in M. persicae.

Research progress of aphid immunity system: Potential effective target for green pest management.

Due to the absence of acquired immunity, insects primarily rely on their innate immune system to resist pathogenic microorganisms and parasitoids in natural habitats. This innate immune system can be classified into cellular immunity and humoral immunity. Cellular immunity is mediated by hemocytes, which perform phagocytosis, aggregation, and encapsulation to fight against invaders, whereas the humoral immunity primarily activates the immune signaling pathways and induces the generation of immune effectors. Existing studies have revealed that the hemipteran aphids lack some crucial immune genes compared to other insect species, indicating the different immune mechanisms in aphids. The current review summarizes the adverse impacts of pathogenic microorganisms and parasitoids on aphids, introduces the cellular and humoral immune systems in insects, and analyzes the differences between aphids and other insect species. Furthermore, our review also discussed the existing prospects and challenges in aphid immunity research, and proposed the potential application of immune genes in green pest management.

Integrated transcriptomic and proteomic analyses reveal the molecular mechanism underlying the thermotolerant response of Spodoptera frugiperda.

Spodoptera frugiperda (Lepidoptera: Noctuidae) is a highly destructive invasive pest with remarkable adaptability to extreme climatic conditions, posing a substantial global threat. Although the effects of temperature stress on the biological and ecological properties of S. frugiperda have been elucidated, the molecular mechanisms underlying its responses remain unclear. Herein, we combined transcriptomic and proteomic analyses to explore the key genes and proteins involved in thermotolerance regulation in S. frugiperda larvae at 42 °C. Overall, 1528 differentially expressed genes (DEGs) and 154 differentially expressed proteins (DEPs) were identified in S. frugiperda larvae under heat stress, including antioxidant enzymes, heat shock proteins (Hsps), cytochrome P450s, starch and sucrose metabolism genes, and insulin signaling pathway genes, indicating their involvement in heat tolerance regulation. Correlation analysis of DEGs and DEPs revealed that seven and eight had the same and opposite expression profiles, respectively. After nanocarrier-mediated RNA interference knockdown of SfHsp29, SfHsp20.4, SfCAT, and SfGST, the body weight and mortality of S. frugiperda larvae significantly decreased and increased under heat stress, respectively. This indicates that SfHsp29, SfHsp20.4, SfCAT, and SfGST play a crucial role in the thermotolerance of S. frugiperda larvae. These results provide insight into the mechanism of heat tolerance in S. frugiperda.

Spatial distribution and core community of diazotrophs in Biological soil crusts and subsoils in temperate semi-arid and arid deserts of China.

Introduction: Biological soil crusts (BSCs) are distributed in arid and semiarid regions, and they function as important microhabitats for nitrogen fixation. The diazotroph community is critical for nitrogen fixation in BSCs and their subsoils. However, little is known about the key groups in different types of BSCs and subsoils in temperate semi-arid or arid deserts. Methods: Here, we sampled three types of BSCs and their subsoils from the Inner Mongolian plateau, investigated the distribution characteristics of the diazotroph community by high-throughput sequencing, predicted keystone species using the molecular ecological network analyses pipeline (MENAP), and verified their close relationship with the available nitrogen (AN) content. Results: The results showed that available nitrogen content in BSCs was higher than that in subsoils in three different types of BSCs, and there were differences among seasons and according to the mean annual precipitation. The abundance of diazotrophs was higher in Cyano-BSCs, while diversity had no significant difference among BSCs and subsoils. Cyanobacteria and Proteobacteria, Nostocaceae and Scytonemataceae, Skermanella, Scytonema, Azohydromonas, Nostoc and Trichormus were the dominant phyla, families, and genera, respectively. The dominant groups belong to Skermanella, Scytonema, and Nostoc formed the core diazotroph community in the three types of BSCs and subsoils, and each had a close relationship with AN. Discussion: These results indicate that diazotrophs in BSCs and subsoils had high diversity, and the core diazotroph communities have a close relationship with nitrogen fixation and that they may be the main contributor to nitrogen fixing in BSCs and subsoils in temperate deserts.

Expression and function of opsin genes associated with phototaxis in Zeugodacus cucurbitae Coquillett (Diptera: Tephritidae).

BACKGROUND: Zeugodacus cucuribitae is a major agricultural pest that causes significant damage to varieties of plants. Vision plays a critical role in phototactic behavior of herbivorous insects. However, the effect of opsin on the phototactic behavior in Z. cucuribitae remains unknown. The aim of this research is to explore the key opsin genes that associate with phototaxis behavior of Z. cucurbitae. RESULTS: Five opsin genes were identified and their expression patterns were analyzed. The relative expression levels of ZcRh1, ZcRh4 and ZcRh6 were highest in 4-day-old larvae, ZcRh2 and ZcRh3 were highest in 3rd-instar larvae and 5-day-old pupae, respectively. Furthermore, five opsin genes had the highest expression levels in compound eyes, followed by the antennae and head, whereas the lower occurred in other tissues. The expression of the long-wavelength-sensitive (LW) opsins first decreased and then increased under green light exposure. In contrast, the expression of ultraviolet-sensitive (UV) opsins first increased and then decreased with the duration of UV exposure. Silencing of LW opsin (dsZcRh1, dsZcRh2, and dsZcRh6) and UV opsin (dsZcRh3 and dsZcRh4) reduced the phototactic efficiency of Z. cucurbitae to green light by 52.27%, 60.72%, and 67.89%, and to UV light by 68.59% and 61.73%, respectively. CONCLUSION: The results indicate that RNAi inhibited the expression of opsin, thereby inhibiting the phototaxis of Z. cucurbitae. This result provides theoretical support for the physical control of Z. cucurbitae and lays the foundation for further exploration of the mechanism of insect phototaxis. © 2023 Society of Chemical Industry.

Molecular characterization and elucidation of the function of Hap38 MAPK in the response of Helicoverpa armigera (Hübner) to UV-A stress.

The cotton bollworm Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae), an important pest of cotton, is detrimental to cotton production. Light from UV-A ultraviolet lamps is regarded as a form of environmental stress for insects. In order to investigate the response of H. armigera exposed to UV-A, we explored Hap38 MAPK expression and functions. We hope that the findings of this study will lay the foundation for future investigations into the insect's phototaxis mechanism. A p38 MAPK was cloned and named Hap38 MAPK. A phylogenetic tree showed that Hap38 MAPK was highly conserved. The gene was highly expressed in the thorax and females. Under UV-A stress, the expression of the gene decreased significantly. After silencing Hap38 MAPK, the activity of the antioxidant enzymes SOD, POD, CAT, and GR decreased. This study suggested that Hap38 MAPK responds to UV-A irradiation and plays critical roles in the defense response to environmental stresses.

Induced heat shock protein 70 confers biological tolerance in UV-B stress-adapted Myzus persicae (Hemiptera).

As an environmental stress factor, ultraviolet-B (UV-B) radiation directly affects insect growth, development, and reproduction. Heat shock protein 70s kDa (Hsp70s) plays an important role in the environmental adaptation of insects. To determine the role of MpHsp70s in the UV-B tolerance of Myzus persicae (Sulzer), we identified the complete complementary DNA sequences of seven MpHsp70s. They were found to be ubiquitously expressed during different developmental stages and were highly expressed in second-instar nymphs and wingless adults. The expression levels of the MpHsp70s were significantly upregulated when exposed to different durations of UV-B stress. Nanocarrier-mediated dsMpHsp70 suppressed the expression of the MpHsp70s and reduced the body length, weight, survival rate, and fecundity of M. persicae under UV-B exposure. When the combinational RNAi approach was adopted, the effects on the survival rate and fecundity were greater under UV-B stress, except for MpHsc70-4. These results suggest that MpHsp70s are essential for the resistance of M. persicae to UV-B stress.

Molecular characterization of six heat shock protein 70 genes from Arma chinensis and their expression patterns in response to temperature stress.

Arma chinensis is an important predatory enemy of many agricultural and forest pests. Heat shock protein 70 (Hsp70) plays an essential role in insect adaptation to various stress factors. To explore the functions of Hsp70s in relation to thermal tolerance of A. chinensis, full-length cDNAs of six Hsp70 genes (AcHsp70Ba, AcHsp70-4, AcHsp68a, AcHsp68b, AcHsp70-2, and AcHsc70-4) were cloned. Their open reading frames (ORFs) were 1902, 2454, 1884, 1905, 1872, and 1947 bp, respectively. Developmental expression profiles showed that AcHsp70Ba, AcHsp70-4, and AcHsc70-4 were extremely highly expressed in adult stages. AcHsp68a and AcHsp70-2 showed the highest level of expression in nymph stages, and AcHsp68b was mainly expressed in male adults. Tissue distribution analysis demonstrated that the AcHsp70s were ubiquitously expressed but showing gene-specific and sex-driven patterns of expression. High temperature induced the expression of the six AcHsp70s. Among them, AcHsp70Ba, AcHsp70-4, AcHsp68a, and AcHsc70-4 were significantly induced at 38 °C for 6 h, while all six AcHsp70s were significantly induced at 38 °C for 24 h. There were differences in responses of the six AcHsp70s to low-temperature stress. The expressions of AcHsp70-4, AcHsp68a, and AcHsp68b in male adults were significantly repressed at 4 °C for 6 h, whereas AcHsp70Ba and AcHsp70-2 were significantly induced. The levels of AcHsp70Ba, AcHsp68b, and AcHsp70-2 in female adults were significantly repressed at 4 °C for 24 h, whereas AcHsc70-4 was significantly induced. These results suggested that AcHsp70s play important roles in various developmental stages and tissue function, and contribute to the tolerance of A. chinensis to extreme temperatures.

Morphological and Molecular Identification of Fusarium ipomoeae as the Causative Agent of Leaf Spot Disease in Tobacco from China.

Tobacco (Nicotiana tabacum L.), which creates jobs for 33 million people and contributes two trillion dollars' tax annually, is one of the most important economic plants globally. However, tobacco is seriously threatened by numerous diseases during production. Previously, the field survey of tobacco diseases was conducted in the Guizhou and Guangxi provinces, the two main tobacco-producing areas in China. A serious leaf spot disease, with a 22% to 35% incidence, was observed in farming plants. In order to determine the causal agents, we collected the disease samples and isolated the pathogenic fungi. The pathogen was identified as Fusariumipomoeae, based on the morphological characteristics and phylogenetic analysis. Pathogenicity tests showed that F. ipomoeae could induce tobacco leaf spot and blight. To our knowledge, this is the first report worldwide of F. ipomoeae causing leaf spots and stems on tobacco. Our study reveals the serious consequences of F. ipomoeae on tobacco filed production and provides information for future diagnosis and management of the Fusarium disease.

Tobacco Root Microbial Community Composition Significantly Associated With Root-Knot Nematode Infections: Dynamic Changes in Microbiota and Growth Stage.

The root-knot nematode (RKN) is an important pathogen that affects the growth of many crops. Exploring the interaction of biocontrol bacteria-pathogens-host root microbes is the theoretical basis for improving colonization and controlling the effect of biocontrol bacteria in the rhizosphere. Therefore, 16S and 18S rRNA sequencing technology was used to explore the microbial composition and diversity of tobacco roots (rhizosphere and endophytic) at different growth stages in typical tobacco RKN-infected areas for 2 consecutive years. We observed that RKN infection changed the α-diversity and microbial composition of root microorganisms and drove the transformation of microorganisms from bacteria to fungi. The abundance of Sphingomonas decreased significantly from 18% to less than 3%, while the abundance of Rhizobiaceae increased from 4 to 15% at the early growth stage during the first planting year, and it promoted the proliferation of Chryseobacterium at the late growth stage in rhizosphere microorganisms with the highest abundance of 17%. The overall trend of rhizosphere microorganisms changed in the early growth stage with increasing growth time. The specific results were as follows: (1) Rhizobiaceae and Chryseobacterium increased rapidly after 75 days, became the main abundant bacteria in the rhizosphere microorganisms. (2) The dominant flora in fungi were Fusarium and Setophoma. (3) Comparing the root microbes in 2017 and 2018, RKN infection significantly promoted the proliferation of Pseudomonas and Setophoma in both the rhizosphere and endophytes during the second year of continuous tobacco planting, increasing the relative abundance of Pseudomonas from 2 to 25%. Pseudomonas was determined to play an important role in plant pest control. Finally, a total of 32 strains of growth-promoting bacteria were screened from tobacco rhizosphere bacteria infected with RKN through a combination of 16S rRNA sequencing and life-promoting tests. The results of this research are helpful for analyzing the relationship between RKNs and bacteria in plants, providing reference data for elucidating the pathogenesis of RKNs and new ideas for the biological control of RKNs. GRAPHICAL ABSTRACT.

Systematic and functional analysis of non-specific lipid transfer protein family genes in sugarcane under Xanthomonas albilineans infection and salicylic acid treatment.

Plant non-specific lipid transfer proteins (nsLTPs) are small basic proteins that play a significant regulatory role in a wide range of physiological processes. To date, no genome-wide survey and expression analysis of this gene family in sugarcane has been performed. In this study we identified the nsLTP gene family in Saccharum spontaneum and carried out expression profiling of nsLTPs in two sugarcane cultivars (Saccharum spp.) that have different resistance to leaf scald caused by Xanthomonas albilineans (Xa) infection. The effect of stress related to exogenous salicylic acid (SA) treatment was also examined. At a genome-wide level, S. spontaneum AP85-441 had 71 SsnsLTP genes including 66 alleles. Tandem (9 gene pairs) and segmental (36 gene pairs) duplication events contributed to SsnsLTP gene family expansion. Five SsnsLTP proteins were predicted to interact with five other proteins. Expression of ShnsLTPI.8/10/Gb.1 genes was significantly upregulated in LCP85-384 (resistant cultivar), but downregulated in ROC20 (susceptible cultivar), suggesting that these genes play a positive regulatory role in response of sugarcane to Xa infection. Conversely, ShnsLTPGa.4/Ge.3 appears to act as a negative regulator in response Xa infection. The majority (16/17) of tested genes were positively induced in LCP85-384 72 h after SA treatment. In both cultivars, but particularly in LCP85-384, ShnsLTPIV.3/VIII.1 genes were upregulated at all time-points, suggesting that the two genes might act as positive regulators under SA stress. Meanwhile, both cultivars showed downregulated ShnsLTPGb.1 gene expression, indicating its potential negative role in SA treatment responses. Notably, the ShnsLTPGb.1 gene had contrasting effects, with positive regulation of gene expression in response to Xa infection and negative regulation induced by SA stress. Together, our results provide valuable information for elucidating the function of ShnsLTP family members under two stressors and identified novel gene sources for development of sugarcane that are tolerant of environmental stimuli.