Population performance and detoxifying and protective enzyme activities of four thrips species feeding on flowers of Magnolia grandiflora (Ranunculales: Magnolia).

BACKGROUND: Different thrips species can co-occur on the same flowers with different dominance degrees. To accurately evaluate the population performance on different thrips species on Magnolia grandiflora flowers, we investigated the diversity of thrips species and their population dynamics both in the field and laboratory. In addition, the activities of detoxifying and protective enzymes in thrips were also measured. RESULTS: Field investigations revealed that four thrips species (Thrips hawaiiensis, Thrips flavidulus, Frankliniella occidentalis, and Thrips coloratus) coexisted on M. grandiflora flowers. They were ranked, from highest population density to lowest, as follows: T. hawaiiensis > T. flavidulus > F. occidentalis > T. coloratus. In laboratory investigations, the species were ranked, from fastest developmental rates to slowest, as follows: F. occidentalis > T. hawaiiensis > T. flavidulus > T. coloratus; and from largest population size to smallest, as follows: T. hawaiiensis > F. occidentalis > T. flavidulus > T. coloratus. Biochemistry assays showed that the four species differed in their activities of detoxifying enzymes (carboxylesterase, glutathione-S-transferase, and cytochrome P450) and protective enzymes (superoxide dismutase, peroxidase) in both laboratory and field strains. CONCLUSION: Differences in population performance among these four thrips on M. grandiflora may be related to their activity levels of physiological enzymes. The variations in thrips population performance between the field and the laboratory could be due to differences in environmental conditions. T. hawaiiensis showed a strong host preference for M. grandiflora, and thus it has the potential to be a dangerous pest in horticultural plants. © 2023 Society of Chemical Industry.

Full-length transcriptome assembly of andrias davidianus (amphibia: caudata) skin via hybrid sequencing.

The Chinese giant salamander, Andrias davidianus, is the largest amphibian species in the world; it is thus an economically and ecologically important species. The skin of A. davidianus exhibits complex adaptive structural and functional adaptations to facilitate survival in aquatic and terrestrial ecosystems. Here, we report the first full-length amphibian transcriptome from the dorsal skin of A. davidianus, which was assembled using hybrid sequencing and the PacBio and Illumina platforms. A total of 153,038 transcripts were hybrid assembled (mean length of 2039 bp and N50 of 2172 bp), and 133,794 were annotated in at least one database (nr, Swiss-Prot, KEGG, KOGs, GO, and nt). A total of 58,732, 68,742, and 115,876 transcripts were classified into 24 KOG categories, 1903 GO term categories, and 46 KEGG pathways (level 2), respectively. A total of 207,627 protein-coding regions, 785 transcription factors, 27,237 potential long non-coding RNAs, and 8299 simple sequence repeats were also identified. The hybrid-assembled transcriptome recovered more full-length transcripts, had a higher N50 contig length, and a higher annotation rate of unique genes compared with that assembled in previous studies using next-generation sequencing. The high-quality full-length reference gene set generated in this study will help elucidate the genetic characteristics of A. davidianus skin and aid the identification of functional skin proteins.

Structural characterization and in vitro immunogenicity evaluation of amphibian-derived collagen type II from the cartilage of Chinese Giant Salamander (Andrias davidianus).

Collagen type II (CT-II) has unique biological activities and functions, yet the knowledge on amphibian-derived CT-II is rare. Herein, acid-soluble collagen (ASC) and pepsin-soluble collagen (PSC) were successfully isolated and characterized from the cartilage of Chinese Giant Salamander (CGS). The in vitro immunogenicity of collagen was then evaluated and compared with that of the standard bovine CT-II (SCT-II) by T-lymphocyte cell proliferation activity. Results demonstrated that ASC and PSC were predominantly CT-II along with minor collagen type I and maintained intact triple-helical structure of nature collagen. Compared with SCT-II, higher glycine content (337.80 and 339.93 residues/1000 residues) and lower degree of proline hydroxylation (51.81% and 52.52%) were observed in ASC and PSC. Additionally, PSC showed comparable Td (63 °C) and higher Tm (109 °C) than SCT-II (64 °C and 103 °C, respectively), indicating its high thermal and structural stability. SEM revealed that the lyophilized ASC and PSC had interconnected porous network structures of collagen-based materials. Moreover, different from SCT-II, both ASC and PSC presented no immunogenicity because they did not cause obvious proliferation of murine T-lymphocyte regardless of the induced concentration of collagen increased from 8 to 417 µg/mL. These data suggested that the amphibian-derived CGS cartilage collagens avoid the immunogenic risk of terrestrial animal collagen, and show high thermal stability and potential advantage in biomedical application.

Identification and Expression Analysis of Four Small Heat Shock Protein Genes in Cigarette Beetle, Lasioderma serricorne (Fabricius).

Small heat shock proteins (sHsps) are molecular chaperones that play crucial roles in the stress adaption of insects. In this study, we identified and characterized four sHsp genes (LsHsp19.4, 20.2, 20.3, and 22.2) from the cigarette beetle, Lasioderma serricorne (Fabricius). The four cDNAs encoded proteins of 169, 180, 181, and 194 amino acids with molecular weights of 19.4, 20.2, 20.3, and 22.2 kDa, respectively. The four LsHsp sequences possessed a typical sHsp domain structure. Quantitative real-time PCR analyses revealed that LsHsp19.4 and 20.3 transcripts were most abundant in pupae, whereas the transcript levels of LsHsp20.2 and 22.2 were highest in adults. Transcripts of three LsHsp genes were highly expressed in the larval fat body, whereas LsHsp20.2 displayed an extremely high expression level in the gut. Expression of the four LsHsp genes was dramatically upregulated in larvae exposed to 20-hydroxyecdysone. The majority of the LsHsp genes were significantly upregulated in response to heat and cold treatments, while LsHsp19.4 was insensitive to cold stress. The four genes were upregulated when challenged by immune triggers (peptidoglycan isolated from Staphylococcus aureus and from Escherichia coli 0111:B4). Exposure to CO2 increased LsHsp20.2 and 20.3 transcript levels, but the LsHsp19.4 transcript level declined. The results suggest that different LsHsp genes play important and distinct regulatory roles in L. serricorne development and in response to diverse stresses.

Fabrication and characterization of Chinese giant salamander skin composite collagen sponge as a high-strength rapid hemostatic material.

Chinese giant salamander (CGS) has high medicinal value and long history of clinical use in ancient China. In this study, CGS skin (CGSS) collagen was extracted and purified to prepare collagen sponge by freeze-drying. TEMPO oxidized microfibrillated cellulose (TEMPO-MFC) and genipin were adopted to improve the mechanical properties of collagen sponge. The hygroscopicity, porosity, mechanical properties, hemostatic performance, morphology, and biodegradability of the resultant sponges were investigated in detail. The results indicated that CGSS collagen was type I collagen with intact triple-helical structure, and the prepared sponge had porous structure and excellent hemostatic performance with procoagulant ratio of 53.28%. However, the CGSS collagen sponge showed low tensile strength (TS) of 98.80 KPa, compression strength (CS) of 1.48 KPa, and elongation at break (E) of 4.72%. Incorporating 2.5% TEMPO-MFC into the native CGSS collagen sponge resulted in an increase of 188.26% in TS to 284.80 KPa, 166.89% in CS to 3.95 KPa, and 73.52% in E to 8.19%. The improvements were attributed to the physical filling of TEMPO-MFC in cavity and cavity wall of collagen sponge and the stable chemical linkage between carboxyl of TEMPO-MFC and amino group of collagen which effectively improved the toughening of sponge and formed good interface bonding, respectively. Subsequent 0.3% genipin treatment further improved the TS to 605.00 KPa and the CS to 8.66 KPa as a result of crosslinking reaction. Moreover, the composite reinforcing also improved the anti-degradation ability and procoagulant ratio of collagen sponge. All results suggested that the TEMPO-MFC toughened and genipin crosslinked CGSS composite collagen sponge is a promising rapid hemostatic material with high-strength and can be applicated in biomedical field.

Functional characterization of chitin deacetylase 1 gene disrupting larval-pupal transition in the drugstore beetle using RNA interference.

Chitin deacetylases (CDAs) are chitin degradation enzymes that strictly regulate growth and development in insects. In this study, we identified and characterized a full-length cDNA of the CDA gene (SpCDA1) in the drugstore beetle, Stegobium paniceum. The open reading frame of SpCDA1 (1614 bp) encoded a 537 amino acid protein, which possessed typical domain structures of CDAs. Phylogenetic comparison to other insect CDAs revealed that SpCDA1 belongs to Group Ib CDAs. Quantitative real-time PCR analyses showed that SpCDA1 was highly expressed in late larval stages. Significant increase of SpCDA1 transcript level in the larvae was observed upon the exposure of 20-hydroxyecdysone. Injection of double-stranded RNA (dsRNA) of SpCDA1 into the late larvae significantly reduced SpCDA1 transcript levels, resulted in larval-pupal molting difficulty and produced high larval mortality. After 15 days, the survival rate of S. paniceum in dsSpCDA1 group was significantly reduced by 72% compared to the control. The results demonstrated that SpCDA1 is essential for successful larval-pupal transition in S. paniceum and this gene may be a potential target for pest control.

Effects of Temperature on the Development and Reproduction of Thrips hawaiiensis (Thysanoptera: Thripidae).

Environmental temperature has a significant impact on insect behavior. The present study aimed to determine the effects of temperature on the development, survival, and reproduction of Thrips hawaiiensis (Thysanoptera: Thripidae), an important flower-inhabiting thrips. These effects were evaluated at five constant temperatures (18, 21, 24, 27, and 30°C) on thrips reared in the laboratory on excised Gardenia jasminoides flowers. The developmental durations of egg, first instar, second instar, prepupa, pupa, and the entire immature stages were shortened in response to a temperature increase from 18 to 30°C. The highest generational survival rate was at 27°C (75.00%), whereas the lowest was at 18°C (46.00%). The minimum threshold and effective accumulated temperatures for completing a generation of T. hawaiiensis were 7.62°C and 171.26 degree-days, respectively. The highest fecundity (95.80) was at 27°C, but it was not significantly different than at 24°C (84.72) or 30°C (84.32). The highest oviposition rate of 5.57 eggs per female per day occurred at 27°C, which was significantly higher than at any other temperature. Both the highest intrinsic rate of increase, at 0.200, and net reproduction rate, at 44.97, for T. hawaiiensis were observed at 27°C, whereas the lowest values of 0.114 and 25.56, respectively, were observed at 18°C. These results suggest that T. hawaiiensis is well adapted to temperate conditions, with an optimal temperature range for development of 24 to 30°C, with the most suitable temperature for both development and reproduction being 27°C.

Complete mitochondrial genome of the bamboo snout beetle, Cyrotrachelus buqueti (Coleoptera: Curculionidae).

The bamboo snout beetle Cyrotrachelus buqueti (Coleoptera: Curculionidae) is a destructive forest pest and distributed widely in Southeast Asia. The 15,035 bp complete mitochondrial genome of the species consists of 13 protein-coding genes (PCGs), two ribosomal RNA genes (rRNAs), 21 transfer RNA genes (tRNAs) and a control region (GenBank accession no. MG674390). The trnl gene was not found in the C. buqueti mitogenome. The gene order and the orientation of C. buqueti were similar to those found in other Coleoptera species. The nucleotide composition was significantly biased (A, G, C, and T was 38.18%, 10.10%, 16.16%, and 35.56%, respectively) with A + T contents of 73.74%. ATG, ATA, ATT, AAT, and TTG were initiation codons and TAA, TAG, and T were termination codons. All the 21 tRNAs displayed a typical cloverleaf secondary structure, except for trnS1 which lacked the dihydrouridine arm. Phylogenetic analysis was performed using 13 PCGs with 14 other beetles showed that C. buqueti is closely related to Eucryptorhynchus brandti, which agree with the traditional classification.

RhMKK9, a rose MAP KINASE KINASE gene, is involved in rehydration-triggered ethylene production in rose gynoecia.

BACKGROUND: Flower opening is an important process in the life cycle of flowering plants and is influenced by various endogenous and environmental factors. Our previous work demonstrated that rose (Rosa hybrida) flowers are highly sensitive to dehydration during flower opening and the water recovery process after dehydration induced ethylene production rapidly in flower gynoecia. In addition, this temporal- and spatial-specific ethylene production is attributed to a transient but robust activation of the rose MAP KINASE6-ACC SYNTHASE1 (RhMPK6-RhACS1) cascade in gynoecia. However, the upstream component of RhMPK6-RhACS1 is unknown, although RhMKK9 (MAP KINASE KINASE9), a rose homologue of Arabidopsis MKK9, could activate RhMPK6 in vitro. In this study, we monitored RhMKK2/4/5/9 expression, the potential upstream kinase to RhMPK6, in rose gynoecia during dehydration and rehydration. RESULTS: We found only RhMKK9 was rapidly and strongly induced by rehydration. Silencing of RhMKK9 significantly decreased rehydration-triggered ethylene production. Consistently, the expression of several ethylene-responsive genes was down regulated in the petals of RhMKK9-silenced flowers. Moreover, we detected the DNA methylation level in the promoter and gene body of RhMKK9 by Chop-PCR. The results showed that rehydration specifically elevated the DNA methylation level on the RhMKK9 gene body, whereas it resulted in hypomethylation in its promoter. CONCLUSIONS: Our results showed that RhMKK9 possibly acts as the upstream component of the RhMKK9-RhMPK6-RhACS1 cascade and is responsible for water recovery-triggered ethylene production in rose gynoecia, and epigenetic DNA methylation is involved in the regulation of RhMKK9 expression by rehydration.

The complete mitochondrial genome of the cigarette beetle, Lasioderma serricorne (Coleoptera: Anobiidae).

The cigarette beetle, Lasioderma serricorne (Fabricius), is an important pest of stored commodities and distributed widely in the world. Here, we report the complete mitochondrial genome of L. serricorne which was 15,958 bp and composed of 13 protein-coding genes (PCGs), two rRNA genes, 22 tRNA genes and a control region. The gene order and orientation of L. serricorne were identical to those of other Coleopteran mitogenomes. ATG, ATA, ATT, ATC, TTG were initiation codons and TAA, TAG, T were termination codons. All 22 tRNA genes were predicted with a typical cloverleaf structure except for trnS1 (AGN). Phylogenetic analysis performed using 13 PCGs with 14 other beetles showed that L. serricorne is closely related to Stegobium paniceum, which agree with the conventional taxonomy.

The complete mitochondrial genome of Stegobium paniceum (Coleoptera: Anobiidae).

The complete mitochondrial genome of Stegobium paniceum (Coleoptera: Anobiidae) is a circular DNA molecule of 15,271 bp (GenBank accession number XK819317), and its nucleotide composition is biased towards A + T nucleotides (78.32%). This genome comprises 13 protein-coding genes, 2 ribosomal RNA genes, 22 transfer RNA (tRNA) genes, and an A + T-rich region. The gene order of S. paniceum was similar to those found in other known Coleoptera species. Sixteen reading frame overlaps and six intergenic regions were found in the mitochondrial genome of S. paniceum. All 22 tRNA genes have the typical cloverleaf secondary structure, with an exception for trnS1 (AGN). The phylogenetic relationships based on neighbour-joining method revealed that S. paniceum is closely related to Apatides fortis, which is consistent with the traditional morphological classification.

Responses of rose RhACS1 and RhACS2 promoters to abiotic stresses in transgenic Arabidopsis thaliana.

KEY MESSAGE: Promoter activities of RhACS1 and RhACS2 , two rose genes involved in ethylene biosynthesis, are highly sensitive to various abiotic stresses in an organ-specific manner. Our previous studies indicated that two rose (Rosa hybrida) 1-aminocyclopropane-1-carboxylic acid synthase genes, RhACS1 and RhACS2, play a role in dehydration-induced ethylene production and inhibition of cell expansion in rose petals. Here, both RhACS1 and RhACS2 promoters were analyzed using histochemical staining and glucuronidase synthase (GUS) gene reporter activity assays following their introduction into transgenic Arabidopsis thaliana plants. It was found that the promoter activities of both genes were strong throughout the course of development from young seedlings to mature flowering plants in various organs, including hypocotyls, cotyledons, leaves, roots and lateral roots. RhACS1 promoter activity was induced by drought in both rosette leaves and roots of transgenic A. thaliana lines, but was reduced following a re-hydration treatment. In contrast, RhACS2 promoter activity was decreased by drought in rosette leaves, while its response pattern was similar to that of RhACS1 in roots. A mannitol treatment induced the activity of both the RhACS1 and RhACS2 promoters, indicating that both genes are also regulated by osmotic stress. In addition, RhACS2 appeared to be abscisic acid (ABA)-inducible, while RhACS1 was less sensitive to ABA. Finally, four truncated sequences of the RhACS1 promoter were generated and GUS activity assays demonstrated that deleting a 327 bp region between bp 862 and -535 resulted in a substantial decrease of the promoter activity. Taken together, our results suggest that the RhACS1 and RhACS2 promoters respond to abiotic stresses in a developmentally regulated and spatially specific manner.

Precise spatio-temporal modulation of ACC synthase by MPK6 cascade mediates the response of rose flowers to rehydration.

Drought is a major abiotic stress that affects the development and growth of most plants, and limits crop yield worldwide. Although the response of plants to drought has been well documented, much less is known about how plants respond to the water recovery process, namely rehydration. Here, we describe the spatio-temporal response of plant reproductive organs to rehydration using rose flowers as an experimental system. We found that rehydration triggered rapid and transient ethylene production in the gynoecia. This ethylene burst serves as a signal to ensure water recovery in flowers, and promotes flower opening by influencing the expression of a set of rehydration-responsive genes. An in-gel kinase assay suggested that the rehydration-induced ethylene burst resulted from transient accumulation of RhACS1/2 proteins in gynoecia. Meanwhile, RhMPK6, a rose homolog of Arabidopsis thaliana MPK6, is rapidly activated by rehydration within 0.5 h. Furthermore, RhMPK6 was able to phosphorylate RhACS1 but not RhACS2 in vitro. Application of the kinase inhibitor K252a suppressed RhACS1 accumulation and rehydration-induced ethylene production in gynoecia, and the protein phosphatase inhibitor okadaic acid had the opposite effect, confirming that accumulation of RhACS1 was phosphorylation-dependent. Finally, silencing of RhMPK6 significantly reduced ethylene production in gynoecia when flowers were subjected to rehydration. Taken together, our results suggest that temporal- and spatial-specific activation of an RhMPK6-RhACS1 cascade is responsible for rehydration-induced ethylene production in gynoecia, and that the resulting ethylene-mediated signaling pathway is a key factor in flower rehydration.

TRV-GFP: a modified Tobacco rattle virus vector for efficient and visualizable analysis of gene function.

Virus-induced gene silencing (VIGS) is a useful tool for functional characterization of genes in plants. Unfortunately, the efficiency of infection by Tobacco rattle virus (TRV) is relatively low for some non-Solanaceae plants, which are economically important, such as rose (Rosa sp.). Here, to generate an easy traceable TRV vector, a green fluorescent protein (GFP) gene was tagged to the 3' terminus of the coat protein gene in the original TRV2 vector, and the silencing efficiency of the modified TRV-GFP vector was tested in several plants, including Nicotiana benthamiana, Arabidopsis thaliana, rose, strawberry (Fragaria ananassa), and chrysanthemum (Dendranthema grandiflorum). The results showed that the efficiency of infection by TRV-GFP was equal to that of the original TRV vector in each tested plant. Spread of the modified TRV virus was easy to monitor by using fluorescent microscopy and a hand-held UV lamp. When TRV-GFP was used to silence the endogenous phytoene desaturase (PDS) gene in rose cuttings and seedlings, the typical photobleached phenotype was observed in 75-80% plants which were identified as GFP positive by UV lamp. In addition, the abundance of GFP protein, which represented the concentration of TRV virus, was proved to correlate negatively with the level of the PDS gene, suggesting that GFP could be used as an indicator of the degree of silencing of a target gene. Taken together, this work provides a visualizable and efficient tool to predict positive gene silencing plants, which is valuable for research into gene function in plants, especially for non-Solanaceae plants.

Transesterification of rapeseed oil for biodiesel production in trickle-bed reactors packed with heterogeneous Ca/Al composite oxide-based alkaline catalyst.

A conventional trickle bed reactor and its modified type both packed with Ca/Al composite oxide-based alkaline catalysts were studied for biodiesel production by transesterification of rapeseed oil and methanol. The effects of the methanol usage and oil flow rate on the FAME yield were investigated under the normal pressure and methanol boiling state. The oil flow rate had a significant effect on the FAME yield for the both reactors. The modified trickle bed reactor kept over 94.5% FAME yield under 0.6 mL/min oil flow rate and 91 mL catalyst bed volume, showing a much higher conversion and operational stability than the conventional type. With the modified trickle bed reactor, both transesterification and methanol separation could be performed simultaneously, and glycerin and methyl esters were separated additionally by gravity separation.

An organ-specific role for ethylene in rose petal expansion during dehydration and rehydration.

Dehydration is a major factor resulting in huge loss from cut flowers during transportation. In the present study, dehydration inhibited petal cell expansion and resulted in irregular flowers in cut roses, mimicking ethylene-treated flowers. Among the five floral organs, dehydration substantially elevated ethylene production in the sepals, whilst rehydration caused rapid and elevated ethylene levels in the gynoecia and sepals. Among the five ethylene biosynthetic enzyme genes (RhACS1-5), expression of RhACS1 and RhACS2 was induced by dehydration and rehydration in the two floral organs. Silencing both RhACS1 and RhACS2 significantly suppressed dehydration- and rehydration-induced ethylene in the sepals and gynoecia. This weakened the inhibitory effect of dehydration on petal cell expansion. ß-glucuronidase activity driven by both the RhACS1 and RhACS2 promoters was dramatically induced in the sepals, pistil, and stamens, but not in the petals of transgenic Arabidopsis. This further supports the organ-specific induction of these two genes. Among the five rose ethylene receptor genes (RhETR1-5), expression of RhETR3 was predominantly induced by dehydration and rehydration in the petals. RhETR3 silencing clearly aggravated the inhibitory effect of dehydration on petal cell expansion. However, no significant difference in the effect between RhETR3-silenced flowers and RhETR-genes-silenced flowers was observed. Furthermore, RhETR-genes silencing extensively altered the expression of 21 cell expansion-related downstream genes in response to ethylene. These results suggest that induction of ethylene biosynthesis by dehydration proceeds in an organ-specific manner, indicating that ethylene can function as a mediator in dehydration-caused inhibition of cell expansion in rose petals.

Effect of calcination temperature on the activity of solid Ca/Al composite oxide-based alkaline catalyst for biodiesel production.

A solid Ca/Al composite oxide-based alkaline catalyst containing Ca(12)Al(14)O(33) and CaO was prepared by chemical synthesis and thermal activation from sodium aluminate solution and calcium hydroxide emulsion. The effect of calcination temperatures ranging from 120 °C to 1000 °C on activity of the catalyst was investigated. The catalyst calcined at 600 °C showed the highest activity with >94% yield of fatty acid methyl esters (i.e. biodiesel) when applied to the transesterification of rapeseed oil at a methanol:oil molar ratio of 15:1 at 65 °C for 3h. Structure and properties of the catalyst were studied and the characterizations with XRD, TGA, FTIR, BET, and SEM demonstrated that the performance of the catalyst was closely related to its specific surface area and crystalline structure. In particular, the generation of crystalline Ca(12)Al(14)O(33) improved the catalytic activity due its synergistic effect with CaO.