Analysis of differences in tobacco leaf microbial communities after redrying in Chinese provinces and from abroad.

Microorganisms play an important role in the tobacco aging process. Before the aging process, raw tobacco leaves must be threshed and redried. In order to explore the differences of microbial community structure of threshed and redried tobacco leaves from different origins at home and abroad, 14 groups of tobacco leaves from 8 different countries were tested by high-throughput DNA sequencing and microbiology analysis. Then, through amplicon sequence variants (ASV) cluster analysis, Venn diagram and species labeling and other microbial diversity analysis, the dominant bacteria and fungi on the surface of threshed and redried tobacco leaves were obtained. The results showed that there were significant differences in the composition of tobacco bacteria and fungi after threshing and redrying from different geographical areas. The relative abundance of Microbacterium and Sphingomonas in domestic tobacco leaves was significantly higher than that of foreign tobacco leaves. The relative abundance of Pseudomonas in foreign tobacco bacterial colonies was significantly higher than that of domestic tobacco leaves. In terms of fungi, the relative abundance of Aspergillus and Alternaria in domestic tobacco leaves was significantly higher than that of foreign tobacco leaves. Septoria, Sampaiozyma, Cladosporium and Phoma account for significantly higher proportions of foreign tobacco leaves. These microorganisms may be indispensable in aging process to form different flavors of tobacco leaves. It provides an important theoretical basis for the further use of microorganisms to promote tobacco leaf aging.

Influence of acid rain and root reinforcement coupling factors on disintegration characteristics of expansive soil.

The effect of soil fixation and anti-scour instability of slope vegetation generally depends on the strength and anti-disintegration ability of slope soil due to increase of root system. Therefore, it is particularly necessary to study the disintegration characteristics of expansive soil related to slope instability under acidic conditions (simulated acid rain). In this paper, the response surface method (RSM) was used with the pH value, root diameter, root length, root coefficient, and distribution as independent variables, and the disintegration amount of root-soil (DARS) after 60min as the response value. Then X-ray diffractometer (XRD) was used to analyze the mineral composition changes of the sample under this environment. Simultaneously, the plasticity index of expansive soil at different values of pH was studied to discuss the disintegration mechanism of root compound expansive soil in an acid environments. The results show that the root system improves the anti-disintegration characteristics of the root-soil, and the effects of various factors on the amount of disintegration were as follows: root length > pH value > root distribution > root amount > root diameter. The DARS with a length of 20mm increased by 26.67% and 41.56% compared to the 30mm and 40mm. Compared to the horizontal distribution and horizontal + slant distribution, the DARS with slant distribution was increases by 11.39% and 20.24% respectively. The DARS with 2 roots is increased by 9.92% and 16.75% compared to 4 and 6 roots respectively. The 1mm diameter DARS is 6.65% and 15.49% higher than the 2mm and 3mm, respectively. In addition, an acidic environments can lead to an increase in the amount of disintegration or rate of disintegration. The disintegration at pH = 4.2 was increased by 11.4% and 22.4% compared to pH = 5.6 and pH = 7, respectively. The acidity affects soil disintegration is due to the hydrophilic minerals in the expansive soil react with H+ ion in the acid solution to form soluble salts. Due to the dissociation and leaching of free quartz and metal oxides in the soil to varying degrees, the ability of expansive soil to accumulate is reduced. The intensity of erosion and leaching decreases with increasing pH. In addition, the pH value can affects the plasticity index of the soil, which increases with the increasing pH, thus affects the disintegration properties of the expansive soil.

A novel victorivirus isolated from the tobacco spot blight fungus Stagonosporopsis cucurbitacearum in China.

Stagonosporopsis cucurbitacearum is an important plant-pathogenic fungus that causes stem and leaf blight diseases in a variety of crops. Here, we report the characterization of a novel victorivirus, tentatively named "Stagonosporopsis cucurbitacearum victorivirus 1" (ScVV-1), isolated from the S. cucurbitacearum isolate M-7. The ScVV-1 genome is 5,165 bp in length with a predicted GC content of 60.1% and contains two large open reading frames (ORF 1 and ORF2) encoding putative proteins that share significant sequence similarity with coat proteins (CPs) and RNA-dependent RNA polymerases (RdRps) of mycoviruses of the family Totiviridae. The ScVV-1 RdRp appears to be translated using a stop-initiation pentanucleotide UAAUG sequence. Phylogenetic analysis based on CP and RdRp amino acid (aa) sequences both indicated that ScVV-1 belongs to the genus Victorivirus in the family Totiviridae. To our knowledge, this is the first full-length genome sequence of a victorivirus infecting S. cucurbitacearum.

Ultrasound-Assisted Extraction of Flavonoids from Potentilla fruticosa L. Using Natural Deep Eutectic Solvents.

A series of natural deep eutectic solvents (NADESs) were prepared with choline chloride, betaine, and a variety of natural organic acids in order to find new environmentally-friendly green solvents to replace the traditional solvents. The NADESs were employed to extract flavonoids from Potentilla fruticosa L. (PFL) with the help of ultrasound. The eutectic solvent diluted with an appropriate amount of water improved the extraction ability of flavonoids due to the decrease of solution viscosity. The microstructure of the raw sample and the samples subjected to ultrasonic bath in different solutions were observed using scanning electron microscope (SEM) to determine the role of the NADESs in the extraction process. The DPPH method and glucose consumption method were used to study the antioxidant and hypoglycemic ability of flavonoid compounds in PFL. Single factor method and response surface methodology (RSM) were designed to analyze the effects of three extraction parameters, including solvent/solid ratio, ultrasonic power, and extraction time, on the extraction yield, antioxidant capacity, and hypoglycemic capacity, and the corresponding second-order polynomial prediction models were established. The optimal extraction conditions for the maximum extraction yield, antioxidant capacity, and hypoglycemic capacity were predicted by RSM, and the reliability of RSM simulation results was verified by a one-off experiment.

Identification and Expression Profiles of Putative Soluble Chemoreception Proteins from Lasioderma serricorne (Coleoptera: Anobiidae) Antennal Transcriptome.

The cigarette beetle, Lasioderma serricorne (Fabricius) (Coleoptera: Anobiidae), is a destructive stored product pest worldwide. Adult cigarette beetles are known to rely on host volatiles and pheromones to locate suitable habitats for oviposition and mating, respectively. However, little is known about the chemosensory mechanisms of these pests. Soluble chemoreception proteins are believed to initiate olfactory signal transduction in insects, which play important roles in host searching and mating behaviors. In this study, we sequenced the antennal transcriptome of L. serricorne and identified 14 odorant-binding proteins (OBPs), 5 chemosensory proteins (CSPs), and 2 Niemann-Pick C2 proteins (NPC2). Quantitative realtime PCR (qPCR) results revealed that several genes (LserOBP2, 3, 6, and 14) were predominantly expressed in females, which might be involved in specific functions in this gender. The five LserOBPs (LserOBP1, 4, 8, 10, and 12) that were highly expressed in the male antennae might encode proteins involved in specific functions in males. These findings will contribute to a better understanding of the olfactory system in this stored product pest and will assist in the development of efficient and environmentally friendly strategies for controlling L. serricorne.

Chromosome-level genome assembly of the mirid predator Cyrtorhinus lividipennis Reuter (Hemiptera: Miridae), an important natural enemy in the rice ecosystem.

Though the genomes of many rice herbivorous pests have recently been well characterized, little is known about the genome of their natural enemies. Here, by using the Illumina and PacBio platforms, we sequenced and assembled the whole genome of the mirid species Cyrtorhinus lividipennis Reuter (Hemiptera: Miridae), which is an economically and ecologically important natural enemy in the rice ecosystem acting as a dominant predator for planthoppers and leafhoppers in the field. Through Hi-C scaffolding, 1615 scaffolds with a total size of 338.08 Mb were successfully anchored onto 13 chromosomes. The assembled genome size was 345.75 Mb with a final scaffold N50 of 27.58 Mb. Approximately 107.51 Mb of sequences accounting for 31.10% of the genome were identified as repeat elements, and 14,644 protein-coding genes were annotated. Phylogenetic analysis showed that C. lividipennis clustered with other Hemipteran species and diverged from Apolygus lucorum about 66.7 million years ago. Gene families related to detoxification, environmental adaptation and digestion were analysed comparatively with other Hemipteran species, but no significant expansion or contraction was found in C. lividipennis. We also observed male meiosis in C. lividipennis, which showed a typical post-reduction of sex chromosomes and a karyotype of 2n = 22 + XY. As the first natural-enemy genome in the rice ecosystem, the genomic resource of C. lividipennis not only expands our understanding of the multitrophic interactions (host plant-prey-predator), but also provides a genomic basis for better understanding this dominant predator and therefore promotes sustainable rice pest management and food grain production.

Diversified Bund Vegetation Coupled With Flowering Plants Enhances Predator Population and Early-Season Pest Control.

Insecticide overuse in crop production systems often results in detrimental effects on predators and parasitoids, which regulate important insect pests. The natural enemies are also unable to survive in monocrop landscapes with the absence of shelter or food sources. Diversified vegetation, especially with flowering plants, can enhance natural enemy abundance and diversity, thus strengthening biological control, enabling farmers to reduce insecticides. In this study, we conserved bund vegetation and manipulated the existing rice landscapes with flowering plants to provide food and shelter for the biological control agents. Our study revealed significant positive relationships between predator densities and bund plant diversity. The abundance of predators significantly increased in the eco-engineered plots, especially at the flowering peaks compared to the insecticide-treated and control plots, while parasitoids were more diverse in both the eco-engineered and control plots. There were no significant differences in planthopper and leafhopper densities among the treatments during the rice early and maximum tillering stages, suggesting effective natural control of these herbivore pests in the eco-engineered plots at the early rice-growing season. However, at the heading stage relatively higher planthopper and leafhopper populations in the control and eco-engineered plots than in the insecticide-sprayed plots were recorded, suggesting perhaps the need for insecticide interventions if exceeding the threshold at this time. Our study indicates that manipulating the habitats surrounding the rice fields to enhance natural enemies is a sustainable practice in rice production as it can enhance the natural suppression of pests and thus reducing the need for insecticide.

Influence of vetiver root on strength of expansive soil-experimental study.

Grassroots have received more attention than the traditional method as soil reinforcement materials, especially the use of vetiver and other vegetation protection methods to treat expansive soil slope, have been tried and applied. To study the influence of grassroots on the strength properties of expansive soil, the laws of vetiver root growth over time and its vertical distribution of root content(δ) were firstly investigated by the experiment of planting vetiver. Then different δ and depth of planted soil were obtained. Simultaneously different δ and water content(ω) of grafted soil were made. With the direct shear test, the shear strength parameters of root-soil with different δ were analyzed. The shear test on root-soil composites with different δ was carried out to compare the strength characteristics of planted and grafted soil. The results showed that the δ of vetiver decreased with the increase of depth, and the δ of each layer increased with the growth period. The δ of 180d was 70.5% higher than that of 90d. The cohesion(c) of root-soil can be increased by more than 97%, and internal friction angle(φ) can be increased by more than 15.4% after 180 days. The c of 90 d vetiver root system can be increased by more than 18%, and the φ can be increased by more than 1.5%. At each depth, the c and φ of composite soil increases with the increase of δ, and the increment of cohesion (Δc) and the increment of internal friction angle (Δφ) increase with the increment of δ. But the increase in the ω will weaken the shear strength parameters of root-soil. Under the condition of the planted root system and grafted root system, the influence degree of δ on strength parameter of root-soil is different, and the law of strength parameters versus δ of grafted soil of 365d is similar to that of planted soil of 90d. And the root reinforcement of grafted soil is weaker than planted soil. Hence the grafted soil can´t accurately reflect the root-soil interaction of the existing root system.

Host Plant Salinity Stress Affects the Development and Population Parameters of Nilaparvata lugens (Hemiptera: Delphacidae).

Salinization is one of the most critical abiotic stress factors for crops and a rising setback in agro-ecosystems. Changes in weather, land usage, and the salinization of irrigation water are increasing soil salinity of many farmlands. Increased soil salinity alters the plant quality, which subsequently may trigger bottom-up effects on herbivorous insect. We examined the bottom-up effect of salinity stress on population parameters of the brown planthopper (BPH), Nilaparvata lugens through rice (Oryza sativa L.) plant. The results revealed that salinity interfered with egg hatching of BPH. The nymphal development period, adult longevity, and oviposition were also influenced by salinity. Notable differences appeared in the intrinsic growth rate (r), the finite increase rate (λ) and the net reproduction rate (R0) of BPH, and a concentration-dependent effect was detected. Although salinity adversely affected BPH development, population projection predicted a successful growth of the BPH population in a relatively short time under the treatment of low and medium levels of salinity (6, 8, and 10 dS/m of NaCl), whereas higher salt concentrations (12 and 14 dS/m) lead to significant fitness costs in BPH populations. Our study predicts that BPH could become a problem in areas with lower and medium salinity and that those planthoppers may exacerbate the negative effects of salinity for rice production. This study will provide valuable information for understanding the field abundance and distribution of BPH on saline rice field, thus contributing to the development of eco-friendly strategies to manage this pest in saline ecosystems.

Identification and expression analysis of putative chemoreception genes from Cyrtorhinus lividipennis (Hemiptera: Miridae) antennal transcriptome.

Cyrtorhinus lividipennis Reuter (Hemiptera: Miridae) is an important egg predator of planthoppers which are destructive rice pests. The chemosensory genes in the mirid antennae play important roles in mating and prey-seeking behaviors. To gain a better understanding of the olfaction of C. lividipennis, we sequenced the antennal transcriptomes of the predator to identify the key olfaction genes. We identified 18 odorant binding proteins (OBPs), 12 chemosensory proteins (CSPs), 1 Niemann-Pick C2 protein (NPC2), 15 odorant receptors (ORs), 6 ionotropic receptors (IRs), 3 gustatory receptors (GRs) and 3 sensory neuron membrane proteins (SNMPs). Quantitative real-time PCR results showed that the relative transcript levels of three ClivORs (ClivOR6, 7 and 14) in the female antennae were 3 to 6 folds higher than that in the male antennae, indicating these genes were more related to oviposition site selection. The relative transcript levels of ClivCSP8 and ClivOR11 were 2.6 and 2.7 times higher in the male antennae than that of the female, respectively, indicating that these genes might be involved in mate searching. Moreover, the responses of dsorco treated predators to volatiles emitted from infested rice were significantly reduced, indicating these volatiles might serve as crucial cues in the host searching of C. lividipennis.

Knockdown of timeless Disrupts the Circadian Behavioral Rhythms in Laodelphax striatellus (Hemiptera: Delphacidae).

Most living organisms developed the innate clock system to anticipate daily environmental changes and to enhance their chances of survival. timeless (tim) is a canonical clock gene. It has been extensively studied in Drosophila melanogaster (Diptera: Drosophilidae) as a key component of the endogenous circadian clock, but its role is largely unknown in some agriculture pests. Laodelphax striatellus (Fallén) (Hemiptera: Delphacidae), an important rice pest, exhibits a robust locomotor rhythm. In the present study, we cloned tim gene (ls-tim) from L. striatellus and investigated its function in the regulation of behavioral rhythms. Quantitative real-time polymerase chain reaction revealed a circadian expression pattern of ls-tim under different light conditions with a trough in the photophase and a peak in the late scotophase. After the knockdown of ls-tim via RNA interference (RNAi), the adults showed an earlier onset of locomotor activity under light/dark cycles and became arrhythmic in constant darkness. ls-tim RNAi also abolished the timing of adult emergence that normally occurs in the early photophase. These results suggest that ls-tim is essential for the light-entrained circadian rhythms in L. striatellus and provide more insights into the endogenous clock network underlying the behavioral and physiological rhythms of this insect.

Cryptochrome Regulates Circadian Locomotor Rhythms in the Small Brown Planthopper Laodelphax striatellus (Fallén).

Most living organisms have developed internal circadian clocks to anticipate the daily environmental changes. The circadian clocks are composed of several transcriptional-translational feedback loops, in which cryptochromes (CRYs) serve as critical elements. In insects, some CRYs act as photopigments to control circadian photoentrainment, while the others act as transcriptional regulators. We cloned and characterized two cryptochrome genes, the Drosophila-like (lscry1) and vertebrate-like (lscry2) genes, in a rice pest Laodelphax striatellus. Quantitative real-time PCR showed that lscry1 and lscry2 expressed ubiquitously from nymph to adult stages as well as in different tissues. The transcript levels of lscry2 fluctuated in a circadian manner. Constant light led to arrhythmic locomotor activities in L. striatellus. It also inhibited the mRNA oscillation of lscry2 and promoted the transcription of lscry1. Knockdown of lscry1 or lscry2 by RNA interference (RNAi) reduced the rhythmicity of L. striatellus in constant darkness, but not in light dark cycles. These results suggested that lscry1 and lscry2 were putative circadian clock genes of L. striatellus, involved in the regulation of locomotor rhythms.

Identification of Candidate Odorant-Binding Protein and Chemosensory Protein Genes in Cyrtorhinus lividipennis (Hemiptera: Miridae), a Key Predator of the Rice Planthoppers in Asia.

Cyrtorhinus lividipennis Reuter (Hemiptera: Miridae) is an important predatory natural enemy of planthopper and leafhopper eggs in Asian rice paddy fields. Cyrtorhinus lividipennis is known to rely largely on herbivore-induced plant volatiles to identify eggs embedded in rice stem tissues for feeding and on pheromones for mating. However, exactly how C. lividipennis decode these chemical information is unclear. In most insects, the odorant-binding proteins (OBPs) and the chemosensory proteins (CSPs) are essential for seeking out food resources and mates. In this study, we identified 10 OBP and 5 CSP genes in C. lividipennis and investigated their expression patterns in various tissues of adult males and females by quantitative real-time PCR (qRT-PCR). Six OBP genes (ClivOBP1, 2, 4, 6, 9, and 10) were mainly expressed in the male antennae, whereas three genes (ClivOBP3, ClivOBP7, and ClivOBP8) had high expression in the female antennae. ClivCSP1 was predominantly expressed in the male antennae. These findings suggest that most ClivOBPs and ClivCSPs are likely involved in food-searching behavior. The recognition of the pheromone molecules provides the basis for further functional studies on the chemoreception system of C. lividipennis.

Odorant-Binding Proteins and Chemosensory Proteins from an Invasive Pest Lissorhoptrus oryzophilus (Coleoptera: Curculionidae).

The rice water weevil, Lissorhoptrus oryzophilus Kuschel (Coleoptera: Curculionidae), is a serious pest species both in its original distribution region of northern America and its invaded regions of eastern Asia and southern Europe. The odorant-binding proteins (OBPs) and the chemosensory proteins (CSPs) play important roles in host and mate locating, thus might play a significant role in the success of the species as an invader, which has not been characterized yet. We identified 10 OBPs and 5 CSPs in L. oryzophilus and investigated the expression profiles of these genes in various tissues by quantitative real-time PCR. Five classic OBPs were predominantly expressed in the antennae. CSPs were expressed ubiquitously with particularly high transcript levels in antennae, legs, and wings. Three antenna-specific OBPs (LoOBP1, 8, 11) were up-regulated following 1-3 d of food deprivation and down-regulated afterward. These findings suggest most classic OBPs are likely involved in chemoreception whereas CSPs as well as the minus-C OBPs may have broader physiological functions, which in turn may help to understand the molecular aspects of chemical communication in this invasive insect.