Physiological changes of Larix principis-rupprechtii seedlings inoculated with Trichoderma spp. under drought stress.

Tree regeneration is significantly affected by water deficiency, which could be alleviated by the inoculation of Trichoderma spp. In this study, the effects of rhizosphere inoculation with Trichoderma spp. on the antioxidant system and osmotic substances in the seedlings of Larix principis-rupprechtii was investigated under experimental drought stresses, with the relative water content 50%-60%, 35%-50%, and 20%-35% as the light, moderate and severe drought stress, respectively. The results showed that, with increasing drought stress, the activities of protective enzymes, SOD and POD in the needles of seedlings were significantly promoted, the contents of MDA and proline were remarkably increased, whereas the contents of soluble protein and starch showed a decreasing trend. Inoculation with Trichoderma spp. promoted much stronger increase in the activities of SOD and POD in the needles, yet the MDA content showed a lower level of increase. Under moderate drought, the activity of POD in the seedling inoculated with Trichoderma spp. was 1.8 folds of that in the control, while the content of MDA was 62.9% of the control. The contents of proline, soluble protein, soluble sugar, and starch in the inoculated seedlings were higher than that in the uninoculated seedlings. Drought-resistance of L. principis-rupprechtii seedlings inoculated by Trichoderma spp. was significantly enhanced due to the augmenting regulation of antioxidant system and osmotic adjustment substances. Therefore, under the trend of warming and drying in North China, the technique of rhizosphere inoculation of Trichoderma spp. could be popularized for forest plantation or understory tending.

Thermal insulation fibers with a Kevlar aerogel core and a porous Nomex shell.

Kevlar aerogel fibers which inherit the aerogel's brilliant properties of low density, high porosity and large surface area are promising candidates for thermal insulation applications in textiles. To enhance the mechanical strength of Kevlar aerogel fibers, an extra Nomex shell was introduced by a simple coaxial-wet-spinning approach. The resultant coaxial fibers were observed to have a Kevlar aerogel core and a porous Nomex shell. Besides, there also formed an air gap between the core and the shell. This multi-layered coaxial structure with numerous pores inside contributes to the excellent thermal insulation performance of the fibers and their fabrics. The temperature differences between the hot plate and the outer surface of the fabrics were measured to be as high as 80 °C when exposed to a temperature of 300 °C. In addition, these fibers also performed well in thermal stability, and almost did not decompose before 380 °C. Not only that, the breaking strength of the Nomex shell can be up to twice that of the Kevlar core, resulting in a significant improvement in the fiber's mechanical strength. It can be envisaged that the developed coaxial fibers with excellent thermal insulation and endurance properties as well as improved mechanical strength may have broad prospects for thermal insulation at high temperatures.

Identification of 48 full-length MHC-DAB functional alleles in miiuy croaker and evidence for positive selection.

Major histocompatibility complex (MHC) molecules play a vital role in the immune response and are a highly polymorphic gene superfamily in vertebrates. As the molecular marker associated with polymorphism and disease susceptibility/resistance, the polymorphism of MHC genes has been investigated in many tetrapods and teleosts. Most studies were focused on the polymorphism of the second exon, which encodes the peptide-binding region (PBR) in the α1- or ß1-domain, but few studies have examined the full-length coding region. To comprehensive investigate the polymorphism of MHC gene, we identified 48 full-length miiuy croaker (Miichthys miiuy) MHC class IIB (Mimi-DAB) functional alleles from 26 miiuy croaker individuals. All of the alleles encode 34 amino acid sequences, and a high level of polymorphism was detected in Mimi-DAB alleles. The rate of non-synonymous substitutions (dN) occurred at a significantly higher frequency than that of synonymous substitutions (dS) in the PBR, and this result suggests that balancing selection maintains polymorphisms at the Mimi-DAB locus. Phylogenetic analysis based on the full-length and exon 2 sequences of Mimi-DAB alleles both showed that the Mimi-DAB alleles were clustered into two major groups. A total of 19 positive selected sites were identified on the Mimi-DAB alleles after testing for positive selection, and 14 sites were predicted to be associated with antigen-binding sites, which suggests that most of selected sites are significant for disease resistance. The polymorphism of Mimi-DAB alleles provides an important resource for analyzing the association between the polymorphism of MHC gene and disease susceptibility/resistance, and for researching the molecular selective breeding of miiuy croaker with enhanced disease resistance.

Characterization of 40 full-length MHC class IIA functional alleles in miiuy croaker: Polymorphism and positive selection.

The major histocompatibility complex is a highly polymorphic gene superfamily in vertebrates that plays an important role in adaptive immune response. In the present study, we identified 40 full-length miiuy croaker MHC class IIA (Mimi-DAA) functional alleles from 26 miiuy croaker individuals and found that the alleles encode 30 amino acid sequences. A high level of polymorphism in Mimi-DAA was detected in miiuy croaker. The rate of non-synonymous substitutions (d(N)) occurred at a significantly higher frequency than that of synonymous substitutions (d(S)) in the peptide-binding region (PBR) and non-PBR. This result suggests that balancing selection maintains polymorphisms at the Mimi-DAA locus. Phylogenetic analysis based on the full-length sequences showed that the Mimi-DAA alleles clustered into three groups. However, the phylogenetic tree constructed using the exon 2 sequences indicated that the Mimi-DAA alleles clustered into two groups. A total of 22 positively selected sites were identified on the Mimi-DAA alleles after testing for positive selection, and five sites were predicted to be associated with the binding of peptide antigen, suggesting that a few selected residues may play a significant role in immune function.

Molecular characterization of three IRF1 subfamily members reveals evolutionary significance of IRF11 in miiuy croaker.

The interferon regulatory factors IRF1 and IRF2 of the IRF1 subfamily play essential roles in immune responses against viruses. IRF11 is a novel IRF gene of the IRF1 subfamily; IRF11 genes share almost the same evolutionary distance with IRF1 and IRF2 genes. However, the structure and characteristics of IRF11 gene in fish have been rarely reported. In our study, IRF1, IRF2 and IRF11 genes were identified and characterized from miiuy croaker genome. Results showed that the IRF1, IRF2 and IRF11 genes contain the same domains; each of these genes is composed of conserved gene organizations and characterized by gene synteny with the orthologous genes. Interestingly, IRF11 was likely found only in fish (but not specific to teleost fish). Evolutionary analysis results showed that IRF1 gene in mammals, IRF2 and IRF11 gene in fish underwent positive selection. IRF1, IRF2 and IRF11 were expressed in a wide range of miiuy croaker tissues. These genes also exhibited the same expression patterns after miiuy croaker was infected with poly(I:C). Therefore, our data enhanced our understanding of the functions and evolution of IRF11 in fish.

Characterization of the miiuy croaker (Miichthys miiuy) transcriptome and development of immune-relevant genes and molecular markers.

BACKGROUND: The miiuy croaker (Miichthys miiuy) is an important species of marine fish that supports capture fisheries and aquaculture. At present commercial scale aquaculture of this species is limited due to diseases caused by pathogens and parasites which restrict production and limit commercial value. The lack of transcriptomic and genomic information for the miiuy croaker limits the ability of researchers to study the pathogenesis and immune system of this species. In this study we constructed a cDNA library from liver, spleen and kidney which was sequenced using Illumina paired-end sequencing to enable gene discovery and molecular marker development. PRINCIPAL FINDINGS: In our study, a total of 69,071 unigenes with an average length of 572 bp were obtained. Of these, 45,676 (66.13%) were successfully annotated in public databases. The unigenes were also annotated with Gene Ontology, Clusters of Orthologous Groups and KEGG pathways. Additionally, 498 immune-relevant genes were identified and classified. Furthermore, 14,885 putative simple sequence repeats (cSSRs) and 8,510 putative single nucleotide polymorphisms (SNPs) were identified from the 69,071 unigenes. CONCLUSION: The miiuy croaker (Miichthys miiuy) transcriptome data provides a large resource to identify new genes involved in many processes including those involved in the response to pathogens and diseases. Furthermore, the thousands of potential cSSR and SNP markers found in this study are important resources with respect to future development of molecular marker assisted breeding programs for the miiuy croaker.

Conserved structural complement component C3 in miiuy croaker Miichthys miiuy and their involvement in pathogenic bacteria induced immunity.

Complement component C3 is a key protein in the complement system whose activation is essential for all the important functions performed by this system. In this study, the complete C3 cDNA sequence was isolated from the miiuy croaker (Miichthys miiuy), which was high similarity to other complement C3. In this study, we report the primary sequence, the tissue expression profile, the polypeptide domain architecture and the phylogenetic analysis of miiuy croaker C3 gene. Rapid amplification of the cDNA ends (RACE) yielded the full open reading frame of this protein (4974 bp), and subsequent analysis indicated that the M. miiuy C3 gene encoded a protein of 1657 amino acids. The deduced amino acid sequence showed that M. miiuy C3 has conserved residues and domains known to be critical for C3 function. Phylogenetic analysis showed that miiuy croaker was most closely related to Epinephelus coioides. Expression analysis showed that C3 was expressed differentially in miiuy croaker tissues, while liver was the main source of C3 expression. Infection of miiuy croaker with Vibrio anguillarum resulted in significant changes expression of C3 gene in the immune-related tissues. These results showed that C3 gene might play an important role in immune mechanisms.

Miiuy croaker transferrin gene and evidence for positive selection events reveal different evolutionary patterns.

Transferrin (TF) is a protein that plays a central role in iron metabolism. This protein is associated with the innate immune system, which is responsible for disease defense responses after bacterial infection. The clear link between TF and the immune defense mechanism has led researchers to consider TF as a candidate gene for disease resistance. In this study, the Miichthys miiuy (miiuy croaker) TF gene (MIMI-TF) was cloned and characterized. The gene structure consisted of a coding region of 2070 nucleotides divided into 17 exons, as well as a non-coding region that included 16 introns and spans 6757 nucleotides. The deduced MIMI-TF protein consisted of 689 amino acids that comprised a signal peptide and two lobes (N- and C-lobes). MIMI-TF expression was significantly up-regulated after infection with Vibrio anguillarum. A series of model tests implemented in the CODEML program showed that TF underwent a complex evolutionary process. Branch-site models revealed that vertebrate TF was vastly different from that of invertebrates, and that the TF of the ancestors of aquatic and terrestrial organisms underwent different selection pressures. The site models detected 10 positively selected sites in extant TF genes. One site was located in the cleft between the N1 and N2 domains and was expected to affect the capability of TF to bind to or release iron indirectly. In addition, eight sites were found near the TF exterior. Two of these sites, which could have evolved from the competition for iron between pathogenic bacteria and TF, were located in potential pathogen-binding domains. Our results could be used to further investigate the function of TF and the selective mechanisms involved.

Complete mitochondrial genome of the red drum, Sciaenops ocellatus (Perciformes, Sciaenidae): absence of the typical conserved motif in the origin of the light-strand replication.

In this study, the complete mitochondrial genome of the red drum Sciaenops ocellatus was determined first. The genome was 16,500 bp in length and contained 13 protein-coding genes, 2 ribosomal RNA genes, 22 transfer RNA genes, and 2 main non-coding regions (the control region and the origin of the light-strand replication); the gene composition and order of which were similar to most other vertebrates. The overall base composition of the heavy strand was T 25.5%, C 30.7%, A 27.5%, and G 16.3%, with a slight AT bias of 53%. Within the control region, the discrete and conserved sequence blocks were identified. Motif 5'-ACCGG-3' rather than 5'-GCCGG-3' was detected in the origin of light-strand replication (O(L)) of red drum, which is rare in the mitogenomes of Sciaenidae species. These results would play an important role in elucidating sequence-function relationships of the O(L).