Molecular and functional characterization of ferritin in abalone Haliotis diversicolor supertexta

Ferritin is an iron storage protein that plays a key role in the processes of physiology and pathology.In the present study,the authors reported the ferritin gene from abalone Haliotis diversicolor supertexta,which we named hds-ferritin.The full-length of hds-ferritin cDNA consisted of 879 bp with an ORF encoding a 171 amino acids.Amino acid sequence analysis revealed that hds-ferritin shared highly homology with other species.Real time PCR and western blot analysis showed that hds-ferritin was distributed ubiquitously in abalone tissues and had the highest expression level in digestive glands,but its transcripts are not modified remarkably by the stimulation with LPS.The recombinant protein was successfully expressed in Escherichia coli BL21 (DE3),and the titre of anti-ferritin antibody was about 1∶14000.The effects of ROS and RNS on ferritin were analyzed in the present study.The results showed that H2O2 played an important role in decreasing hds-ferritin,however NO cation appeared to have a protecting effect on H2O2-medied reduction of hds-ferritin.

Genome size evolution of the extant lycophytes and ferns

Ferns and lycophytes have remarkably large genomes.However,little is known about how their genome size evolved in fern lineages.To explore the origins and evolution of chromosome numbers and genome size in ferns,we used flow cytometry to measure the genomes of 240 species(255 samples)of extant ferns and lycophytes comprising 27 families and 72 genera,of which 228 species(242 samples)represent new reports.We analyzed correlations among genome size,spore size,chromosomal features,phylogeny,and habitat type preference within a phylogenetic framework.We also applied ANOVA and multinomial logistic regression analysis to preference of habitat type and genome size.Using the phylogeny,we conducted ancestral character reconstruction for habitat types and tested whether genome size changes simultaneously with shifts in habitat preference.We found that 2 C values had weak phylogenetic signal,whereas the base number of chromosomes(x)had a strong phylogenetic signal.Furthermore,our analyses revealed a positive correlation between genome size and chromosome traits,indicating that the base number of chromosomes(x),chromosome size,and polyploidization may be primary contributors to genome expansion in ferns and lycophytes.Genome sizes in different habitat types varied significantly and were significantly correlated with habitat types;specifically,multinomial logistic regression indicated that species with larger 2 C values were more likely to be epiphytes.Terrestrial habitat is inferred to be ancestral for both extant ferns and lycophytes,whereas transitions to other habitat types occurred as the major clades emerged.Shifts in habitat types appear be followed by periods of genomic stability.Based on these results,we inferred that habitat type changes and multiple whole-genome duplications have contributed to the formation of large genomes of ferns and their allies during their evolutionary history.

Characterization and expression analysis of an allograft inflammatory factor-1 homologue in yellow grouper (Epinephelus awoara)

Allograft inflammatory factor-1 （ AIF-1 ） is a cytoplasmic calcium-binding protein involved in inflammatory response-related diseases in mammals. Previously an identified AIF-1 gene was simply reported in yellow grouper. The characterization of AIF-1 gene and its expression at the gene and protein level are further described. Yellow grouper AIF-1 is composed of 147 amino acids, and 64% ~ 84% identical to other homologues. Basal level AIF-1 mRNA expression was noted in spleen, anterior kidney and kidney, using reverse-transcription polymerase chain reaction （RT-PCR）. After stimulation of LPS, the AIF-1 mRNA expression was up-regulated in tissues examined： spleen, anterior kidney, kidney, heart and liver, but not in muscle. The recombinant AIF-1 protein was expressed in Escherichia coli, and then purified for the development of antiserum. Western blotting analysis revealed a band with a molecular mass of about 17 ku.

Screening and optimization of EPA-producing antarctic psychrophilic bacterium Shewanella sp. NJ136

Two hundred strains of bacteria from Antarctic sea ice were collected and screened for their ability of producing eieosapentaenoic acid （EPA, 20： 5ω3） by means of Gas Chromatography/Mass Spetmmetry （GC/MS）. Eight strains of bacteria containing EPA were investigated, among which the outstanding one was recorded as NJ136. This bacterium was identified as Shewanella by the biochemical characteristics and 16S rRNA sequence. Response surface methodology （RSM） was applied to optimize the medium ingredients. A 24 full factorial central composite design （FFCCD） was employed to determine the naximum EPA production at optimum levels of pH, NaC1, glucose and yeast extract. The predicted optimal combination of media constituents for maximum 14.02 mg/g （ about 1.7-fold increase） EPA production were determined as 30.15%e （m/v） NaC1, 9.98 g/L glucose, 4.42 g/L yeast extract and pH 6.08. The actual experimental results were in agreement with the prediction.

Screening on DHA-producing Antarctic bacteria N-6 and its cultural conditions

In Antarctic, the geography and climate differs from those in other places, and the baeteria there have adapted well to the environment there. Two hundred strains of bacteria were isolated from the sea ice in Antarctica. The bacteria were sereened for DHA by means of GC, with fish oil as the standard. Seven strains eontaining DHA or EPA were obtained, among whieh the strain of No. N-6 was outstanding. And the eomponent of DHA was identified by CC-MS. The relative eontent of DHA in N-6 was 8.72%, and total lipids in dry bacteria was 22.54%. The effects of some faetors, ineluding temperature, salinity and pH value, on the growth and DHA-content of strain N-6 were studied. The results show that the DHA-content is relatively high when in low temperature and high pH, and the bacterium is psyehrophilic, alkalophilie.

Relationship between body composition of Zhuang-Dong ethnic groups and natural geographical factors

In order to discuss the law of the change of body composition of Zhuang-Dong ethnic groups with the change of natural geographical factors(longitude,latitude,annual average temperature and annual average sunshine).Using BIA to measure the body composition data of 5098 cases of 13 ethnic groups of Zhuang-Dong group in China.At the same time,collect the relevant data of longitude,latitude,annual average temperature and annual average sunshine of 13 ethnic groups,and use per capita disposable income as a control variable to perform partial correlation analysis about body composition with longitude,latitude,annual average temperature,and annual average illumination.Through this research,it was found Zhuang-Dong group fat rate decrease and little change in muscle mass,presumption of bone mass,and water content with the increase of longitude.

Insights into cryptic speciation of quillworts in China

Cryptic species are commonly misidentified because of high morphological similarities to other species.One group of plants that may harbor large numbers of cryptic species is the quillworts(Isoetes spp.),an ancient aquatic plant lineage.Although over 350 species of Isoetes have been reported globally,only ten species have been recorded in China.The aim of this study is to better understand Isoetes species diversity in China.For this purpose,we systematically explored the phylogeny and evolution of Isoetes using complete chloroplast genome(plastome)data,spore morphology,chromosome number,genetic structure,and haplotypes of almost all Chinese Isoetes populations.We identified three ploidy levels of Isoetes in Chinaddiploid(2n=22),tetraploid(2n=44),and hexaploid(2n=66).We also found four megaspore and microspore ornamentation types in diploids,six in tetraploids,and three in hexaploids.Phylogenetic analyses confirmed that I.hypsophila as the ancestral group of the genus and revealed that Isoetes diploids,tetraploids,and hexaploids do not form monophyletic clades.Most individual species possess a single genetic structure;however,several samples have conflicting positions on the phylogenetic tree based on SNPs and the tree based on plastome data.All 36 samples shared 22 haplotypes.Divergence time analysis showed that I.hypsophila diverged in the early Eocene(~48.05 Ma),and most other Isoetes species diverged 3-20 Ma.Additionally,different species of Isoetes were found to inhabit different water systems and environments along the Yangtze River.These findings provide new insights into the relationships among Isoetes species in China,where highly similar morphologic populations may harbor many cryptic species.

Lycophyte transcriptomes reveal two whole-genome duplications in Lycopodiaceae:Insights into the polyploidization of Phlegmariurus

Lycophytes are an ancient clade of the non-flowering vascular plants with chromosome numbers that vary from tens to hundreds.They are an excellent study system for examining whole-genome duplications(WGDs),or polyploidization,in spore-dispersed vascular plants.However,a lack of genome sequence data limits the reliable detection of very ancient WGDs,small-scale duplications(SSDs),and recent WGDs.Here,we integrated phylogenomic analysis and the distribution of synonymous substitutions per synonymous sites(Ks)of the transcriptomes of 13 species of lycophytes to identify,locate,and date multiple WGDs in the lycophyte family Lycopodiaceae.Additionally,we examined the genus Phlegmariurus for signs of genetic discordance,which can provide valuable insight into the underlying causes of such conflict(e.g.,hybridization,incomplete lineage sorting,or horizontal gene transfer).We found strong evidence that two WGD events occurred along the phylogenetic backbone of Lycopodiaceae,with one occurring in the common ancestor of extant Phlegmariurus(Lycopodiaceae)approximately 22-23 million years ago(Mya)and the other occurring in the common ancestor of Lycopodiaceae around 206-214 Mya.Interestingly,we found significant genetic discordance in the genus Phlegmariurus,indicating that the genus has a complex evolutionary history.This study provides molecular evidence for multiple WGDs in Lycopodiaceae and offers phylogenetic clues to the evolutionary history of Lycopodiaceae.

Using DNA damage to monitor water environment

DNA damage of aquatic organisms living in polluted environments can be used as a biomarker of the genotoxicity of toxic agents to organisms. This technique has been playing an important role in ecotoxi- cological study and environmental risk assessment. In this article, main types of DNA damage caused by pollut- ants in water environments were reviewed; methods of detecting DNA damage were also documented for water environmental monitoring.

Rickettsia-like organism infection associated with mass mortalities of blood clam, Tegillarca granosa, in the Yueqing Bay in China

A series of mass mortalities of the cultured blood clam, Tegillarca granosa, occurred in the Yueqing Bay of China from 2005 to 2009. An obligate intracellular prokaryote, designated as rickettsia-like organism （RLO）, was frequently found in the moribund or dead blood clam sample during ultra- structural examination. These organisms were usually round, ellipsoid or occasionally dumbbell- shaped, ranged from approximately 0.28 to 0.71 #m in size and had a trilaminar cell wall. Two reproductive modes of organisms, transverse binary fission and budding, were observed. The or- ganisms were able to form eosinophilic inclusions. Most inclusions were found within epithelial and connective tissues of the mantle, gills and digestive tube. The biological and morphological char- acteristics indicate that these organisms may belong to the family Rickettsiaceae. RLOs exhibited significant pathogenicity. Cytopathological examinations revealed extensive necrosis and destruc- tion in the infected cell. The degree of tissue destruction was positively related to the number of RLO inclusions in the tissues, and the cytopathological effects were positively related to the number of intracellular RLO. RLOs and their inclusions were discovered throughout different disease areas and in different time periods. The infection intensity of the RLOs was positively correlated with the mortality rate of clams. Therefore, RLO infection might be associated with mass mortalities of cultured blood clams in the Yueqing Bay.

Determination on Blood Physiological Parameters of Apodemus agrarius

The study was conducted to investigate the characteristics of blood physiological parameters of Apodemus agrarius （A. agrarius）. Blood physiological pa- rammers of 10 A. agrarius （half male and half female） were determined by automated hematology analyzer. Results showed that difference of A. agrarius hematokrit （HCT） between male and female was significant （P 〈 0.05 ）, and differences of all the other blood physiological parameters between male and female were not sig- nificant （ P 〉 0.05 ）. In the study, blood physiological reference value of A. agrarius was set up, which provided a basis for the bio-research and prophylaxis and treatment of A. agrarius.

Decomposition Method of Genetic Correlation Coefficient Based on NC Ⅱ Mating Design

There are different degrees of correlation between crop traits. The phenotypic correlation is decomposed into genetic and environmental correlation in quantitative genetics. In this paper,according to stochastic model of variance and covariance analysis,we calculate different genetic components,bring up a decomposition method of genetic correlation coefficient based on NC II mating design,and use examples to show analytic steps and interpret results.

Improvement in bioconversion efficiency and reduction of ammonia emission by introduction of fruit fermentation broth in a black soldier fly larvae and kitchen waste conversion system

The black soldier fly(BSF),Hermetia illucens(Diptera:Stratiomyidae),is an insect commonly used for the bioconversion of various organic wastes.Not only can the BSF convert organic waste into macromolecular organic substances,such as insect pro-teins,but it can also lessen the pollution associated with these waste products by reducing ammonia emissions,for example.In this study,we measured the effects of adding fruit fermentation broth(Fer)and commercial lactic acid bacteria fermentation broth(Em)to kitchen waste(KW),as deodorizing auxiliary substances,on the growth performance of black soldier fly larvae(BSFL),the intestinal flora structure of BSFL,the ammonia emis-sion from the KW substrate,and the microbial community structure of the KW substrate.We found that the addition of Fer or Em increased the body weight of BSFL after 6 d of culture,increasing the growth rate by 9.96%and 7.96%,respectively.The addition of Fer not only reduced the pH of the KW substrate but also increased the relative abundance of probiotics,such as Lactobacillus,Lysinibacillus,and Vagococcus,which inhibited the growth of ammonifiers such as Bacillus,Oligella,Paenalcaligenes,Paenibacillus,Pseu-dogracilibacillus,and Pseudomonas,resulting in the reduction of ammonia emission in the KW substrate.Moreover,the addition of Fer or Em significantly increased the rela-tive abundances of Bacteroides,Campylobacter,Dysgonomonas,Enterococcus,and Ig-natzschineria in the gut of BSFL and increased the species diversity and richness in the K W substrate.Our findings provide a novel way to improve the conversion rate of organic waste and reduce the environmental pollution caused by BSF.

Systemically functional characterization of regiospecific flavonoid O-methyltransferases from Glycine max

Plants produce diverse flavonoids for defense and stress resistance,most of which have health benefits and are widely used as food additives and medicines.Methylation of the free hydroxyl groups of flavonoids,catalyzed by S-adenosyl-l-methionine-dependent O-methyltransferases(OMTs),significantly affects their physicochemical properties and bioactivities.Soybeans(Glycine max)contain a rich pool of O-methylated flavonoids.However,the OMTs responsible for flavonoid methylation in G.max remain largely unknown.We screened the G.max genome and obtained 22 putative OMT-encoding genes that share a broad spectrum of amino acid identities(25–96%);among them,19 OMTs were successfully cloned and heterologously expressed in Escherichia coli.We used the flavonoids containing the free 3,5,7,8,3′,4′hydroxyl group,such as flavones(luteolin and 7,8-dihydroxyflavone),flavonols(kaempferol and quercetin),flavanones(naringenin and eriodictyol),isoflavonoids(daidzein and glycetein),and caffeic acid as substrates,and 15 OMTs were proven to catalyze at least one substrate.The methylation activities of these GmOMTs covered the 3,7,8,3′,4′-hydroxyl of flavonoids and 7,4′-hydroxyl of isoflavonoids.The systematic characterization of G.max flavonoid OMTs provides insights into the biosynthesis of methylated flavonoids in soybeans and OMT bioparts for the production of methylated flavonoids via synthetic biology.

Functional Analysis of Nodulin-like Promoter in Transgenic Cotton Plants

For the first time, a nodulin-like gene promoter was isolated from Gossypium hirsutum L. Guo Y 18 by means of inverse PCR. Three plant expression vectors were constructed for functional identification of the promoter. These vectors were different only in promoter regions; three truncations of the nodulinlike promoter took the place of the CaMV35S promoter in the pBI 121 plant expression vector. Then, the three vectors were introduced into cotton plants via the pollen tube pathway. The expression patterns of the gus gene driven by nodulin-like promoter truncations were investigated in the offspring of transgenic cotton plants. Histochemical GUS staining and fluorescence quantitative analysis were performed to achieve this goal. The results showed that the nodulin-like promoter was a strong, highly reproductive organspecific promoter, which demonstrated a much higher driver activity than the CaMV35S promoter did in cotton reproductive organs, but relatively lower activity in vegetation. Identification of the speciality and strength-determining regions of the nodulin-like promoter was also undertaken.

Synthetic biology of plant natural products: From pathway elucidation to engineered biosynthesis in plant cells

Plant natural products(PNPs)are the main sources of drugs,food additives,and new biofuels and have become a hotspot in synthetic biology.In the past two decades,the engineered biosynthesis of many PNPs has been achieved through the construction of microbial cell factories.Alongside the rapid development of plant physiology,genetics,and plant genetic modification techniques,hosts have now expanded from single-celled microbes to complex plant systems.Plant synthetic biology is an emerging field that combines engineering principles with plant biology.In this review,we introduce recent advances in the biosynthetic pathway elucidation of PNPs and summarize the progress of engineered PNP biosynthesis in plant cells.Furthermore,a future vision of plant synthetic biology is proposed.Although we are still a long way from overcoming all the bottlenecks in plant synthetic biology,the ascent of this field is expected to provide a huge opportunity for future agriculture and industry.

Elucidation of the melitidin biosynthesis pathway in pummelo

Specialized plant metabolism is a rich resource of compounds for drug discovery.The acylated flavonoid glycoside melitidin is being developed as an anti-cholesterol statin drug candidate,but its biosynthetic route in plants has not yet been fully characterized.Here,we describe the gene discovery and functional characterization of a new flavonoid gene cluster(UDP-glucuronosyltransferases(Cg UGTs),1,2rhamnosyltransferase(Cg1,2Rha T),acyltransferases(Cg ATs))that is responsible for melitidin biosynthesis in pummelo(Citrus grandis(L.)Osbeck).Population variation analysis indicated that the tailoring of acyltransferases,specific for bitter substrates,mainly determine the natural abundance of melitidin.Moreover,3-hydroxy-3-methylglutaryl-Co A reductase enzyme inhibition assays showed that the product from this metabolic gene cluster,melitidin,may be an effective anti-cholesterol statin drug candidate.Co-expression of these clustered genes in Nicotiana benthamiana resulted in the formation of melitidin,demonstrating the potential for metabolic engineering of melitidin in a heterologous plant system.This study establishes a biosynthetic pathway for melitidin,which provides genetic resources for the breeding and genetic improvement of pummelo aimed at fortifying the content of biologically active metabolites.