A tomato chloroplast-targeted DnaJ protein, SlDnaJ20 maintains the stability of photosystem I/II under chilling stress.

DnaJ proteins are key molecular chaperones that act as a part of the stress response to stabilize plant proteins, thereby maintaining protein homeostasis under stressful conditions. Herein we used transgenic plants to explore the role of the tomato (Solanum lycopersicum) SlDnaJ20 chloroplast DnaJ protein in to the resistance of these proteins to cold. When chilled, transgenic plants exhibited superior cold resistance, with reduced growth inhibition and cellular damage and increased fresh mass and chlorophyll content relative to control. These transgenic plants further exhibited increased Fv/Fm, P700 oxidation, φRo, and δRo relative to control plants under chilling conditions. Under these same cold conditions, these transgenic plants also exhibited higher levels of core proteins in the photosystem I (PSI) and II (PSII) complexes (PsaA and PsaB; D1 and D2) relative to control wild-type plants. Together these results suggested that the overexpression of SlDnaJ20 is sufficient to maintain PSI and PSII complex stability and to alleviate associated photoinhibition of these complexes, thereby increasing transgenic plant resistance to cold stress.

The Molecular Structure and Self-Assembly Behavior of Reductive Amination of Oxidized Alginate Derivative for Hydrophobic Drug Delivery.

On account of the rigid structure of alginate chains, the oxidation-reductive amination reaction was performed to synthesize the reductive amination of oxidized alginate derivative (RAOA) that was systematically characterized for the development of pharmaceutical formulations. The molecular structure and self-assembly behavior of the resultant RAOA was evaluated by an FT-IR spectrometer, a 1H NMR spectrometer, X-ray diffraction (XRD), thermal gravimetric analysis (TGA), a fluorescence spectrophotometer, rheology, a transmission electron microscope (TEM) and dynamic light scattering (DLS). In addition, the loading and in vitro release of ibuprofen for the RAOA microcapsules prepared by the high-speed shearing method, and the cytotoxicity of the RAOA microcapsules against the murine macrophage RAW264.7 cell were also studied. The experimental results indicated that the hydrophobic octylamine was successfully grafted onto the alginate backbone through the oxidation-reductive amination reaction, which destroyed the intramolecular hydrogen bond of the raw sodium alginate (SA), thereby enhancing its molecular flexibility to achieve the self-assembly performance of RAOA. Consequently, the synthesized RAOA displayed good amphiphilic properties with a critical aggregation concentration (CAC) of 0.43 g/L in NaCl solution, which was significantly lower than that of SA, and formed regular self-assembled micelles with an average hydrodynamic diameter of 277 nm (PDI = 0.19) and a zeta potential of about -69.8 mV. Meanwhile, the drug-loaded RAOA microcapsules had a relatively high encapsulation efficiency (EE) of 87.6 % and good sustained-release properties in comparison to the drug-loaded SA aggregates, indicating the good affinity of RAOA to hydrophobic ibuprofen. The swelling and degradation of RAOA microcapsules and the diffusion of the loaded drug jointly controlled the release rate of ibuprofen. Moreover, it also displayed low cytotoxicity against the RAW264.7 cell, similar to the SA aggregates. In view of the excellent advantages of RAOA, it is expected to become the ideal candidate for hydrophobic drug delivery in the biomedical field.

Fabrication and Evaluation of Alginate/Bacterial Cellulose Nanocrystals-Chitosan-Gelatin Composite Scaffolds.

It is common knowledge that pure alginate hydrogel is more likely to have weak mechanical strength, a lack of cell recognition sites, extensive swelling and uncontrolled degradation, and thus be unable to satisfy the demands of the ideal scaffold. To address these problems, we attempted to fabricate alginate/bacterial cellulose nanocrystals-chitosan-gelatin (Alg/BCNs-CS-GT) composite scaffolds using the combined method involving the incorporation of BCNs in the alginate matrix, internal gelation through the hydroxyapatite-d-glucono-δ-lactone (HAP-GDL) complex, and layer-by-layer (LBL) electrostatic assembly of polyelectrolytes. Meanwhile, the effect of various contents of BCNs on the scaffold morphology, porosity, mechanical properties, and swelling and degradation behavior was investigated. The experimental results showed that the fabricated Alg/BCNs-CS-GT composite scaffolds exhibited regular 3D morphologies and well-developed pore structures. With the increase in BCNs content, the pore size of Alg/BCNs-CS-GT composite scaffolds was gradually reduced from 200 µm to 70 µm. Furthermore, BCNs were fully embedded in the alginate matrix through the intermolecular hydrogen bond with alginate. Moreover, the addition of BCNs could effectively control the swelling and biodegradation of the Alg/BCNs-CS-GT composite scaffolds. Furthermore, the in vitro cytotoxicity studies indicated that the porous fiber network of BCNs could fully mimic the extracellular matrix structure, which promoted the adhesion and spreading of MG63 cells and MC3T3-E1 cells on the Alg/BCNs-CS-GT composite scaffolds. In addition, these cells could grow in the 3D-porous structure of composite scaffolds, which exhibited good proliferative viability. Based on the effect of BCNs on the cytocompatibility of composite scaffolds, the optimum BCNs content for the Alg/BCNs-CS-GT composite scaffolds was 0.2% (w/v). On the basis of good merits, such as regular 3D morphology, well-developed pore structure, controlled swelling and biodegradation behavior, and good cytocompatibility, the Alg/BCNs-CS-GT composite scaffolds may exhibit great potential as the ideal scaffold in the bone tissue engineering field.

Identification and preliminary characterization of chemosensory-related proteins in the gall fly, Procecidochares utilis by transcriptomic analysis.

Chemoreception is critical for insect behaviors such as foraging, host searching and oviposition. The process of chemoreception is mediated by a series of proteins, including odorant-binding proteins (OBPs), gustatory receptors (GRs), odorant receptors (ORs), ionotropic receptors (IRs), chemosensory proteins (CSPs) and sensory neuron membrane proteins (SNMPs). The tephritid stem gall fly, Procecidochares utilis Stone, is a type of egg parasitic insect, which is an effective biological control agent for the invasive weed Ageratina adenophora in many countries. However, the study of molecular components related to the olfactory system of P. utilis has not been investigated. Here, we conducted the developmental transcriptome (egg, first-third instar larva, pupa, female and male adult) of P. utilis using next-generation sequencing technology and identified a total of 133 chemosensory genes, including 40 OBPs, 29 GRs, 24 ORs, 28 IRs, 6 CSPs, and 6 SNMPs. The sequences of these candidate chemosensory genes were confirmed by BLAST, and phylogenetic analysis was performed. Quantitative real-time PCR (qRT-PCR) confirmed that the expression levels of the candidate OBPs varied at the different developmental stages of P. utilis with most OBPs expressed mainly in the pupae, female and male adults but scarcely in eggs and larvae, which was consistent with the differentially expressed genes (DEGs) analysis using the fragments per kilobase per million fragments (FPKM) value. Our results provide a significant contribution towards the knowledge of the set of chemosensory proteins and help advance the use of P. utilis as biological control agents.

Population Numbers and Physiological Response of an Invasive and Native Thrip Species Following Repeated Exposure to Imidacloprid.

Frankliniella occidentalis and F. intonsa are devastating pest insects that target Rosa rugosa, Chrysanthemum morifolium, and Phaseolus vulgaris, which are important economical horticultural plants in China. Meanwhile, R. rugosa and C. morifolium are important cash plants in Kunming, South China. We focus on the population performance of these two thrips species on these three host plants with or without repeated exposure to imidacloprid in Kunming. In the field, the population numbers of F. occidentalis developed faster and were larger on these three sampled host plants, especially under imidacloprid exposure, compared with F. intonsa. The activity of the detoxifying enzymes (CarE, AchE, and MFO) and the antioxidant enzymes (CAT and POD) in both thrips species were significantly enhanced under imidacloprid exposure, whereas the activities of SOD in both thrips were significantly decreased on these three host plants, compared with the control. Overall, enzyme activity of F. occidentalis showed a greater increase than that observed in F. intonsa in most cases, which could be exploited in further studies on thrips resistance management.

Midgut microbiota diversity of potato tuber moth associated with potato tissue consumed.

BACKGROUND: The potato tuber moth (PTM), Phthorimaea operculella (Zeller), is a worldwide pest that feeds on both the leaves and tubers of potato plants. PTM larvae can digest leaves, or tubers, resulting in serious damage to potato plants in the field and potato tubers in storage. To understand how midgut bacterial diversity is influenced by the consumption of these two tissue types, the symbiotic bacteria in the potato-feeding PTM midgut and the endophytic bacteria of potato tissues were analyzed. RESULTS: At the genus level, the bacterial community composition in the PTM midgut was influenced by the tissues consumed, owing to their different nutrient contents. Escherichia_Shigella and Enterobacter were the most dominant genera in the midgut of leaf-feeding and tuber-feeding PTMs, respectively. Interestingly, even though only present in low abundance in leaves and tubers, Escherichia_Shigella were dominantly distributed only in the midgut of leaf-feeding PTMs, indicating that specific accumulation of these genera have occurred by feeding on leaves. Moreover, Enterobacter, the most dominant genus in the midgut of tuber-feeding PTMs, was undetectable in all potato tissues, indicating it is gut-specific origin and tuber feeding-specific accumulation. Both Escherichia_Shigella and Enterobacter abundances were positively correlated with the dominant contents of potato leaves and tubers, respectively. CONCLUSIONS: Enrichment of specific PTM midgut bacterial communities was related to different nutrient levels in different tissues consumed by the insect, which in turn influenced host utilization. We provide evidence that a portion of the intestinal microbes of PTMs may be derived from potato endophytic bacteria and improve the understanding of the relationship between potato endophytic bacteria and the gut microbiota of PTMs, which may offer support for integrated management of this worldwide pest.

The Time-Concentration-Mortality Responses of Western Flower Thrips, Frankliniella occidentalis, to the Synergistic Interaction of Entomopathogenic Fungus Metarhizium flavoviride, Insecticides, and Diatomaceous Earth.

Western flower thrips (WFT), Frankliniella occidentalis (Pergande), is a highly invasive pest which is harmful to many cash crops globally and resistant to various insecticides. Entomopathogenic fungi (EPF), as biological control agents, have demonstrated a good control effect on WFT. The aim of this study was to evaluate the synergistic and pathogenicity efficacy of the fungal strain Metarhizium flavoviride WSWL51721 when distributed with diatomaceous earth (DE) and the active ingredient imidacloprid using four bioassay methods against adult and second instar larvae of WFT. The data of the four bioassays have been fitted to the time-concentration-mortality (TCM) model. The corrected mortality ranges of WFT adults were 75-100%, 82.69-100%, 78.85-100%, and 92.31-100%, and the corrected mortality ranges of WFT second instar larvae were 72.22-100%, 85.19-100%, 77.77-100%, and 100% in the four bioassays at concentrations of 1.2 × 106 to 1.2 × 108 conidia/mL, respectively. At 1.2 × 108 conidia/mL, assays 2 (M. flavoviride with DE), 3 (M. flavoviride with imidacloprid), and 4 (M. flavoviride with DE and imidacloprid) had the shortest median lethal time (LT50), compared with that of assay 1 (M. flavoviride alone) for adults at 2.26 d, 2.06 d, and 1.53 d, and second instar larvae at 2.45 d, 1.70 d, and 1.41 d, respectively. The median lethal concentration (LC50) in the four bioassays decreased within 3-10 days of inoculation. On the third day, it was found that the lowest median lethal concentrations in assays 2, 3, and 4 were 1.58 × 107, 1.13 × 107, and 3.39 × 106 conidia/mL, respectively, which were significantly different from that in assay 1 for the adults. For the second instar larvae, assays 2, 3, and 4 also had the lowest lethal concentrations and were significantly different from those of assay 1. There were significant differences in sporulation between adults and second instar larvae under the four bioassays. Our results indicate that assays 2 (M. flavoviride with DE), 3 (M. flavoviride with imidacloprid), and 4 (M. flavoviride with DE and imidacloprid) demonstrate synergistic effects on the control of both adult and second instar larvae of WFT under laboratory conditions.

De novo transcriptome analysis and identification of genes associated with immunity, detoxification and energy metabolism from the fat body of the tephritid gall fly, Procecidochares utilis.

The fat body, a multifunctional organ analogous to the liver and fat tissue of vertebrates, plays an important role in insect life cycles. The fat body is involved in protein storage, energy metabolism, elimination of xenobiotics, and production of immunity regulator-like proteins. However, the molecular mechanism of the fat body's physiological functions in the tephritid stem gall-forming fly, Procecidochares utilis, are still unknown. In this study, we performed transcriptome analysis of the fat body of P. utilis using Illumina sequencing technology. In total, 3.71 G of clean reads were obtained and assembled into 30,559 unigenes, with an average length of 539 bp. Among those unigenes, 21,439 (70.16%) were annotated based on sequence similarity to proteins in NCBI's non-redundant protein sequence database (Nr). Sequences were also compared to NCBI's non-redundant nucleotide sequence database (Nt), a manually curated and reviewed protein sequence database (SwissProt), and KEGG and gene ontology annotations were applied to better understand the functions of these unigenes. A comparative analysis was performed to identify unigenes related to detoxification, immunity and energy metabolism. Many unigenes involved in detoxification were identified, including 50 unigenes of putative cytochrome P450s (P450s), 18 of glutathione S-transferases (GSTs), 35 of carboxylesterases (CarEs) and 26 of ATP-binding cassette (ABC) transporters. Many unigenes related to immunity were identified, including 17 putative serpin genes, five peptidoglycan recognition proteins (PGRPs) and four lysozyme genes. In addition, unigenes potentially involved in energy metabolism, including 18 lipase genes, five fatty acid synthase (FAS) genes and six elongases of very long chain fatty acid (ELOVL) genes, were identified. This transcriptome improves our genetic understanding of P. utilis and the identification of a numerous transcripts in the fat body of P. utilis offer a series of valuable molecular resources for future studies on the functions of these genes.

Two genera of Panchaetothripinae newly recorded from China, with first description of males of Opimothrips tubulatus (Thysanoptera, Thripidae).

The subfamily Panchaetothripinae comprises 40 extant genera worldwide (ThripsWiki 2019), although only 15 genera are previously recorded from China (Mirab-balou et al. 2016; Li et al. 2018). Panchaetothripinae thrips are characterized by the strong reticulations on the body and leg. Wilson (1975) recognized three tribes in this subfamily, Panchaetothripini, Monilothripini and Tryphactothripini, but only tribe Tryphactothripini was relatively supported based on morphological characters (Mound et al. 2001). Species in this Tribe have abdominal segment II constricted at the base and bearing laterally patches of strong ridges, wart-like tubercles or stoutly recurved microtrichia, and abdominal segment X tends to be asymmetrical. Recently, two further genera of Tryphactothripini were found in Southern China, Noathrips and Opimothrips. These two monotypic genera are reported only from the Old World tropics, Noathrips from India and Sri Lanka, and Opimothrips only from Thailand (Bhatti 1967; Kudô 1979; Nonaka Okajima 1992). The purpose of this paper is to record these two genera from China, together with the first description of the male of Opimothrips tubulatus.

Trichromothrips genus-group (Thysanoptera, Thripidae) from China, with descriptions of three new species and ten new records.

With the addition here of Laplothrips and Trachynotothrips, six genera of Trichromothrips genus-group are now known from China. Newly recorded from China are ten species of this group described originally from various Asian countries, together with three new species, Trichromothrips antidesmae sp.n., T. guizhouensis sp.n. and T. moundi sp.n.. The male of T. elegans is described, and a key provided to the 16 species of Trichromothrips known from China.

A new species and two new records of Rhamphothrips (Thysanoptera: Thripidae) from Southwestern China.

Rhamphothrips dalbergiae sp.n. is described, from the leaves of Dalbergia sp. in Southwestern China. This is the fourth bicoloured species of Rhamphothrips. Tergites VII-VIII of males have discal setae S1 and S2 long and prominent. Two species described from India, R. aureus (Ananthakrishnan) and R. santokhi Kulshretha Vijay Veer, are reported newly from China.

A new Panchaetothripinae genus and species, also a newly recorded genus, from Southwestern China (Thysanoptera: Thripidae).

Araliacothrips daweishanensis gen. et sp.n. is described from Southwestern China in the Thripidae, Panchaetothripinae. In the form of the pronotum and fore wing, and the sculpture on the tergites, this species resembles species of Helionothrips. It is referred to a new genus because the metathoracic endofurca is simple and transverse, the head has cheeks parallel but constricted behind eyes without an occipital apodeme or collar, and tergites III-VIII have a pair of sub-median curved or sigmoid setae. A morphologically similar genus, Aoratothrips, is recorded from China for the first time.

Two new species of Mycterothrips (Thysanoptera: Thripidae) from Southwestern China.

Mycterothrips gongshanensis sp.n. and M. yelangi sp.n. are described from Southwestern China. Both species have been collected from tea (Camellia sinensis), and M. gongshanensis sp.n. is a potential pest in tea gardens. An identification key is provided to the Mycterothrips species recorded from China.

Molecular characterization of the hippeastrum chlorotic ringspot virus L segment and its protein.

Hippeastrum chlorotic ringspot virus (HCRV) is a novel tospovirus that was identified in Yunnan Province, China, in 2013. We have sequenced the HCRV L gene, which is 8909 nt long and encodes the RNA-dependent RNA polymerase (2873 amino acids, 330.8 kDa). The HCRV L protein shared highest similarity (89.4 %) with that of tomato yellow ring virus. The L protein contains a negative-sense RNA virus RNA-directed RNA polymerase motif and an endonuclease domain at the N-terminus. Combined with our previous reports of the S and M RNAs, the genome sequence of HCRV is now completed.

[Dynamics and combined injuries of main pest species in rice cropping zones of Yunnan, Southwest China].

A series of rice pest injuries (due to pathogens, insects, and weeds) were surveyed in 286 farmers' fields for major rice varieties of three rice cropping zones of Yunnan Province, Southwest China. The composition and dynamics of main pest species were analyzed, and the trend of rice pest succession in Yunnan was discussed based upon landmark publications. The results showed that the three rice cropping zones had different pest characteristics as regard to main species, dynamics and combined injuries. Sheath rot, bacterial leaf blight, rice stripe, leaf hoppers, armyworms and stem borers were serious in the japonica rice zone. Sheath blight and rice stripe were serious in the japonica-indica interlacing zone. Leaf blast, sheath blight, leaf folders and weeds above rice crop canopy were serious in the indica rice zone. False smut, plant hoppers and weeds below rice crop canopy were ubiquitous and serious in the three kinds of rice cropping zones. Many kinds of weed infestation emerged in the whole rice cropping seasons. Echinochloa crusgalli, Sagittaria pygmaea, Potamogeton distinctus and Spirodela polyrhiza were the main species of weeds in the rice cropping zones of Yunnan. Overall, levels of combined injuries due to pests in the japonica rice zone and the indica rice zone were higher than that in the japonica-indica interlacing zone. In terms of the trend of rice pest succession in Yunnan, injuries due to false smut, sheath blight and plant hoppers seemed to be in a worse tendency in all rice cropping zones of Yunnan, while dominants species of weeds in the paddy fields are shifting from the annual weeds to the perennial malignant weeds.

[Root growth characteristics and competitive effects of Ageratina adenophora and four functional type herbaceous plants].

A field experiment was conducted to explore the root competitive effects of Ageratina adenophora and Setaria sphacelata, S. yunnanensis, Eupatorium fortunei, Chenopodium serotinum in monoculture and mixture, and the relative competitive abilities were evaluated. The results showed that the root length, superficial area and volume of A. adenophora in mixture were lower than in monoculture, but those of S. sphacelata were higher in mixture than in monoculture. The biomass of A. adenophora in mixture decreased by 77.1% and that of S. sphacelata increased by 80.4% compared with those in monoculture. The relative yield and competitive balance index of S. sphacelata were significantly higher than those of A. adenophora, and the relative yield was about 1.0, suggesting that the underground competitive ability of S. sphacelata was higher than A. adenophora. The root morphology of S. yunnanensis in monoculture and mixture was higher than those of A. adenophora, but the root morphology of two species in mixture was lower than in monoculture. The biomass of A. adenophora and S. yunnanensis in mixture decreased by 45.3% and 22.8% compared with those in monoculture, respectively. Competition effect parameters showed that A. adenophora was a mutual antagonism with S. yunnanensis. The root morphology of E. fortunei and A. adenophora in mixture showed no significant difference compared with that in monoculture. The biomass of A. adenophora and E. fortunei was lower than that in monoculture, respectively. Competition effect parameters showed that A. adenophora was a superior competitor. In the mixture of A. adenophora and C. serotinum, the root morphology parameters and competitive ability of A. adenophora were superior to those of C. serotinum. Above all, S. sphacelata is a preference plant material to control the A. adenophorum invasion and recover biodiversity in A. adenophorum invasion fields.

Identification of a Novel TECTA mutation in a Chinese DFNA8/12 family with prelingual progressive sensorineural hearing impairment.

Tectorial membrane, an extracellular matrix of the cochlea, plays a crucial role in the transmission of sound to the sensory hair cells. Alpha-tectorin is the most important noncollagenous component of the tectorial membrane and the otolith membrane in the maculae of the vestibular system. Defects in TECTA, the gene encodes alpha-tectorin, are cause of both dominant (DFNA8/12) and recessive (DFNB21) forms of deafness. Here, we report a three-generation Chinese family characterized by prelingual progressive sensorineural hearing impairment. We mapped the disease locus to chromosome 11q23-24 region, overlapping with the DFNA8/12 locus. Sequencing of candidate gene TECTA revealed a heterozygous c.5945C>A substitution in exon 19, causing amino acid substitution of Ala to Asp at a conservative position 1982. The A1982D substitution is consistent with hearing loss in this Chinese family and has not been found in 200 random control chromosomes. To our knowledge, this is the first TECTA mutation identified in Chinese population. Our data provides additional molecular and clinical information for establishing a better genotype-phenotype understanding of DFNA8/12.

Quantification of major royal jelly protein 1 in fresh royal jelly by indirect enzyme-linked immunosorbent assay.

Rabbit polyclonal antibody produced by a major royal jelly protein 1 (MRJP1) specific peptide reacted only with a MRJP1. Indirect ELISA with the antibody revealed a MRJP1 level of 4.12-4.67 g/100 g in different company's royal jelly, which almost agreed with that of a hexametric form of MRJP1 (apisin) measured by high performance liquid chromatography. These results suggest that MRJP1 exists mainly as apisin in royal jelly.

[Clinical and genetic features of a large Chinese family with nonsyndromic autosomal dominant hearing loss].

OBJECTIVE: To investigate the clinical and genetical characteristics of a Chinese family with an autosomal-dominant inherited high-frequency sensorineural hearing loss. METHOD: Pedigree was drawn after investigation. Fifeteen family members were checked up, and detailed audiological examination was performed. RESULT: The proband of the kindred had been diagnosed with senserineural hearing loss. A Chinese family SX-G087 with non-sysdromic hearing loss was ascertained. The inheritance pattern of this family is autosomal dominant based on the investigated information. The affected members showed postlingual, progressive, bilateral moderate to severe sensorineural hearing impairment. The age of onset varied from 20 to 35 years. The hearing loss began at high frequencies, and lower frequencies became involved with increasing age. CONCLUSION: Pedigree analysis suggested an autosomal-dominant inheritance pattern in this family. The information should facilitate linkage analysis and positional cloning for the causative gene of this family.

[Characteristics of audiology and clinical genetics of a Chinese family with the DFNA5 genetic hearing loss].

OBJECTIVE: To analysis the characteristics of audiology and clinical genetics of a Chinese family with the DFNA5 genetic hearing loss in detail. METHOD: A detailed family history and clinical data were collected. The Chinese pedigree is an autosomal-dominant inherited hearing loss. The data of audiological examination about genetic characteristics was analysed. The relationship between the hearing-impaired of this family and age was contrasted. RESULT: This Chinese family spanned five generations and comprised 42 members. The mode of inheritance of the families should be autosomal dominant according to the pedigree. Pure-tone audiograms showed a so-called Z shape curve. The hearing loss is sensorineural, progressive and beginning at the high frequencies. The audiograms were fairly symmetric. Whole frequencies became involved with increasing age. CONCLUSION: The Chinese family with the DFNA5 mutation was an autosomal dominant pedigree. In this family, non-syndromic symmetric hearing impairment was severest at the high frequencies early, and gradually accumulated all frequencies of hearing. A mutation in DFNA5 leads to a type of hearing loss that closely resembles the frequently observed age-related hearing impairment. It should take into account DFNA5 mutation which the audiogram of a genetic hearing impaired has the same feature.