丽金龟亚科(Rutelinae)部分种类16S rRNA-ND1基因序列分析

将测得的丽金龟亚科（Rutelinae）7个样本的16S rRNA-ND1基因部分序列与GenBank数据库中的11条同源序列进行比较,应用DNAStar5.0和Mega 3.1软件,分析核苷酸序列组成和遗传距离,构建分子系统发育树。结果表明：在获得的810bp序列长度中,变异位点218个,简约信息位点155个;A＋T使用表现明显的偏向性（73.3%）。丽金龟亚科9个属间的遗传距离在0.054~0.152之间变动;属内种间的距离为0.021~0.064。中华弧丽金龟（Popillia quadriguttata）和日本弧丽金龟（Popillia japonica）具有十分相近的亲缘关系。以白星花金龟（Potosia brevitarsis）和太异爪犀金龟（Heteronychus lioderes）为外群重建的NJ、ME树均支持丽金龟亚科中9属16个种分为2个分支格局的结果 ：原喙丽金龟属（Prodoretus）、鳞喙丽金龟属（Lepadoretus）、喙丽金龟属（Adoretus）聚为一支;弧丽金龟属（Popillia）、藜丽金龟属（Blitopertha）、发丽金龟属（Phyllopertha）、彩丽金龟属（Mimela）、塞丽金龟属（Anisoplia）分化为第二分支。其中,异丽金龟属（Anomala）的不同种优先聚集在该属分支下,证实基于分子分类与形态分类的一致性。

Impact of Different Rates of Nitrogen Supplementation on Soil PhysicochemicalProperties and Microbial Diversity in Goji Berry

Goji berry(Lycium barbarum L.)is substantially dependent on nitrogen fertilizer application,which can signifi-cantly enhance fruit yield and Goji berry industrial development in Ningxia,China.This study aimed to analyze the functions of differential nitrogen application rates including low(N1),medium(N2),and high(N3)levels in soil microbial community structure(bacterial and fungal)at 2 diverse soil depths(0-20,20-40 cm)through high-throughput sequencing technology by targeting 16S RNA gene and ITS1&ITS2 regions.All the observed physicochemical parameters exhibited significant improvement(p<0.05)with increased levels of nitrogen and the highest values for most parameters were observed at N2.However,pH decreased(p<0.05)gradually.The alpha and beta diversity analyses for bacterial and fungal communities’metagenome displayed more similarities than differences among all groups.The top bacterial and fungal phyla and genera suggested no obvious(p>0.05)differences among three group treatments(N1,N2,and N3).Furthermore,the functional enrichment analysis demonstrated significant(p<0.05)enrichment of quorum sensing,cysteine and methionine metabolism,and transcriptional machinery for bacterial communities,while various saprotrophic functional roles for fungal communities.Conclusively,moderately reducing the use of N-supplemented fertilizers is conducive to increasing soil nitrogen utilization rate,which can contribute to sustainable agriculture practices through improved soil quality,and microbial community structure and functions.

Assessing the relationship between the rumen microbiota and feed efficiency in Nellore steers

Background:Ruminants rely upon a complex community of microbes in their rumen to convert host-indigestible feed into nutrients.However,little is known about the association between the rumen microbiota and feed efficiency traits in Nellore(Bos indicus)cattle,a breed of major economic importance to the global beef market.Here,we compare the composition of the bacterial,archaeal and fungal communities in the rumen of Nellore steers with high and low feed efficiency(FE)phenotypes,as measured by residual feed intake(RFI).Results:The Firmicutes to Bacteroidetes ratio was significantly higher(P<0.05)in positive-RFI steers(p-RFI,low feed efficiency)than in negative-RFI(n-RFI,high feed efficiency)steers.The differences in bacterial composition from steers with high and low FE were mainly associated with members of the families Lachnospiraceae,Ruminococcaceae and Christensenellaceae,as well as the genus Prevotella.Archaeal community richness was lower(P<0.05)in p-RFI than in n-RFI steers and the genus Methanobrevibacter was either increased or exclusive of p-RFI steers.The fungal genus Buwchfawromyces was more abundant in the rumen solid fraction of n-RFI steers(P<0.05)and a highly abundant OTU belonging to the genus Piromyces was also increased in the rumen microbiota of highefficiency steers.However,analysis of rumen fermentation variables and functional predictions indicated similar metabolic outputs for the microbiota of distinct FE groups.Conclusions:Our results demonstrate that differences in the ruminal microbiota of high and low FE Nellore steers comprise specific taxa from the bacterial,archaeal and fungal communities.Biomarker OTUs belonging to the genus Piromyces were identified in animals showing high feed efficiency,whereas among archaea,Methanobrevibacter was associated with steers classified as p-RFI.The identification of specific RFI-associated microorganisms in Nellore steers could guide further studies targeting the isolation and functional characterization of rumen microbes potentially important for the energy-harvesting efficiency of ruminants.

Integrating a DNA barcoding project with an ecological survey:a case study on temperate intertidal polychaete communities in Qingdao,China

In this study,we integrated a DNA barcoding project with an ecological survey on intertidal polychaete communities and investigated the utility of CO1 gene sequence as a DNA barcode for the classification of the intertidal polychaetes.Using 16S rDNA as a complementary marker and combining morphological and ecological characterization,some of dominant and common polychaete species from Chinese coasts were assessed for their taxonomic status.We obtained 22 haplotype gene sequences of 13 taxa,including 10 CO1 sequences and 12 16S rDNA sequences.Based on intra-and inter-specific distances,we built phylogenetic trees using the neighbor-joining method.Our study suggested that the mitochondrial CO1 gene was a valid DNA barcoding marker for species identification in polychaetes,but other genes,such as 16S rDNA,could be used as a complementary genetic marker.For more accurate species identification and effective testing of species hypothesis,DNA barcoding should be incorporated with morphological,ecological,biogeographical,and phylogenetic information.The application of DNA barcoding and molecular identification in the ecological survey on the intertidal polychaete communities demonstrated the feasibility of integrating DNA taxonomy and ecology.

Interaction of the major inflammatory bowel disease susceptibility alleles in Crohn’s disease patients

AIM:To investigate the interaction of interleukin-23 receptor(IL23R)(rs1004819 and rs2201841),autophagy-related 16-like 1(ATG16L1)(rs2241880), caspase recruitment domain-containing protein 15 (CARD15)genes,and IBD5 locus in Crohn's disease(CD) patients. METHODS:A total of 315 unrelated subjects with CD and 314 healthy controls were genotyped.Interactions and specific genotype combinations of a total of eight variants were tested.The variants of IBD5locus(IGR2198a_1 rs11739135 and IGR2096a_1 rs12521868),CARD15(R702W rs2066845 and L1007fs rs2066847),ATG16L1(rs2241880)and IL23R (rs1004819,rs2201841)genes were genotyped by PCR-RFLP,the G908R(rs2066844)in CARD15 was determined by direct sequencing. RESULTS:The association of ATG16L1 T300A with CD was confirmed[P=0.004,odds ratio(OR)=1.69, 95%CI:1.19-2.41],and both IL23R variants were found to represent significant risk for the disease(P= 0.008,OR=2.05,95%CI:1.20-3.50 for rs1004819 AA;P<0.001,OR=2.97,95%CI:1.65-5.33 for rs2201841 CC).Logistic regression analysis of pairwise interaction of the inflammatory bowel disease (IBD)loci indicated that IL23R,ATG16L1,CARD15 and IBD5(IGR2198a_1)contribute independently to disease risk.We also analysed the specific combina- tions by pair of individual ATG16L1,IL23R rs1004819, rs2201841,IGR2198a_1,IGR2096a_1 and CARD15 genotypes for disease risk influence.In almost all cases,the combined risk of susceptibility pairs was higher in patients carrying two different risk-associated gene variants together than individuals with just one polymorphism.The highest OR was found for IL23R rs2201841 homozygous genotype with combination of positive CARD15 status(P<0.001,OR=9.15,95% CI:2.05-40.74). CONCLUSION:The present study suggests a cumulative effect of individual IBD susceptibility loci.