Design of tandem genes cluster for isoflavone engineering

In this paper,a universal effective novel method of constructing tandem genes cluster for isoflavone engineering was reported.A tandem genes cluster CHS-CHI-IFS(rIFS)of secondary metabolites of plant isoflavones was constructed by using the chalcone synthase(CHS),chalcone isomerase(CHI),and isoflavone synthase(IFS)(GenBank accession numbers EU526827,EU526829,EU526830)in only one recombination with the pET22b vector.The resulting expression vector pET-rIFS was heterogeneously expressed and co-incubated with barrenwort extractions,and the genistein-like com-ponent was detected.

Associations between IL-1RN variable number of tandem repeat, IL-1β (-511) and IL-1β (+3954) gene polymorphisms and urolithiasis in Uighur children of China

Objective:Interleukin-1(IL-1)is a pro-inflammatory cytokine which may be related to urolithiasis.Genetic polymorphisms of the interleukin-1beta(IL-1β)have been proposed as markers for urolithiasis in some areas.Due to the high incidence of urolithiasis in Uighur children(Xinjiang,China)and existence of ethnic difference,our aim is to explore the potential of IL-1 gene polymorphisms and urolithiasis among these children.Methods:Genomic DNA extracted from peripheral blood of 115 patients and 98 controls were used for genotype polymorphisms analyses.IL-1 receptor antagonist(IL-1RN)gene variable number of tandem repeat(VNTR)gene polymorphisms were analyzed by PCR method.PCR-based restriction analysis was done for the IL-1β(-511)and IL-1β(+3954)gene polymorphisms by endonucleases Ava I and Taq I,respectively.The genotype distribution,allele frequencies,carriage rate,and haplotype frequencies were statistically analyzed.Results:No significant differences were observed in genotypic frequencies between pediatric urolithiasis patients and control group for IL-1RN gene(χ^(2)=1.906,p=0.605),IL-1β(-511)gene(χ^(2)=0.105,p=0.949),or IL-1β(+3954)gene(χ^(2)=3.635,p=0.169).There were yet no significant differences of the allele frequencies of IL-1RN VNTR gene(p=0.779),IL-1β(-511)gene(p=0.941),and IL-1β(+3954)gene(p=0.418)in the case and control groups,as well as the carriage rate and haplotype of them(all p>0.05).Conclusions:The associations between IL-1RN VNTR,IL-1β(-511)and IL-1β(+3954)genes polymorphisms and urolithiasis were not significant in Uighur children.The results need to be confirmed in studies with larger population sample size,as well as in other ethnic groups.

Double-barreled defense: dual ent-miltiradiene synthases in most rice cultivars

Rice(Oryza sativa)produces numerous diterpenoid phytoalexins that are important in defense against pathogens.Surprisingly,despite extensive previous investigations,a major group of such phytoalexins,the abietoryzins,were only recently reported.These aromatic abietanes are presumably derived from ent-miltiradiene,but such biosynthetic capacity has not yet been reported in O.sativa.While wild rice has been reported to contain such an enzyme,specifically ent-kaurene synthase-like 10(KSL10),the only characterized ortholog from O.sativa(OsKSL10),specifically from the well-studied cultivar(cv.)Nipponbare,instead has been shown to make ent-sandaracopimaradiene,precursor to the oryzalexins.Notably,in many other cultivars,OsKSL10 is accompanied by a tandem duplicate,termed here OsKSL14.Biochemical characterization of OsKLS14 from cv.Kitaake demonstrates that this produces the expected abietoryzin precursor ent-miltiradiene.Strikingly,phylogenetic analysis of OsKSL10 across the rice pan-genome reveals that from cv.Nipponbare is an outlier,whereas the alleles from most other cultivars group with those from wild rice,suggesting that these also might produce ent-miltiradiene.Indeed,OsKSL10 from cv.Kitaake exhibits such activity as well,consistent with its production of abietoryzins but not oryzalexins.Similarly consistent with these results is the lack of abietoryzin production by cv.Nipponbare.Although their equivalent product outcome might suggest redundancy,OsKSL10 and OsKSL14 were observed to exhibit distinct expression patterns,indicating such differences may underlie retention of these duplicated genes.Regardless,the results reported here clarify abietoryzin biosynthesis and provide insight into the evolution of rice diterpenoid phytoalexins.

Chromosome-scale genomes of Quercus sichourensis and Quercus rex provide insights into the evolution and adaptation of Fagaceae

The Fagaceae,a plant family with a wide distribution and diverse adaptability,has garnered significant interest as a subject of study in plant speciation and adaptation.Meanwhile,certain Fagaceae species are regarded as highly valuable wood resources due to the exceptional quality of their wood.In this study,we present two high-quality,chromosome-scale genome sequences for Quercus sichourensis(848.75 Mb)and Quercus rex(883.46 Mb).Comparative genomics analysis reveals that the difference in the number of plant disease resistance genes and the nonsynonymous and synonymous substitution ratio(Ka/Ks)of protein-coding genes among Fagaceae species are related to different environmental adaptations.Interestingly,most genes related to starch synthesis in the investigated Quercoideae species are located on a single chromosome,as compared to the outgroup species,Fagus sylvatica.Furthermore,resequencing and population analysis of Q.sichourensis and Q.rex reveal that Q.sichourensis has lower genetic diversity and higher deleterious mutations compared to Q.rex.The high-quality,chromosome-level genomes and the population genomic analysis of the critically endangered Q.sichourensis and Q.rex will provide an invaluable resource as well as insights for future study in these two species,even the genus Quercus,to facilitate their conservation.

Complete mitochondrial genome of Nanorana pleskei （Amphibia： Anura： Dicroglossidae） and evolutionary characteristics

The complete mitochondrial genome of Nanorana pleskei from the Qinghai-Tibet Plateau was sequenced. It includes 17,660 base pairs, containing 13 protein-coding genes, two rRNAs and 23 tRNAs. A tandem duplication of tRNAu^t gene was found in this mitochondrial genome, and the similarity between the two tRNAMet genes is 85.8%, being the highest in amphibian mitochondrial genomes sequenced thus far. Based on gene organization, 24 types were found from 145 amphibian mitochondrial genomes. Type 1 was present in 108 species, type 11 in 11 species, types 5, 16, 17, and 20 each in two species, and the others each present in one species. Fifteen types were found in Anura, being the most diversity in three orders of the Lissamphibia. Our phylogenetic results using 11 protein-coding gene sequences of 145 amphibian mitochondrial genomes strongly support the mo- nophyly of the Lissamphibia, as well as its three orders, the Gymnophiona, Caudata, and Anura, among which the relationships were （（Gymnophiona （Caudata, Anura））. Based on the phylogenetic trees, type 1 was recognized as the ancestral type for am- phibians, and type 11 was the synapomorphic type for the Neobatrachia. Gene rearrangements among lineages provide meaning- ful phylogenetic information. The rearrangement of the LTPF tRNA gene cluster and the translocation of the ND5 gene only found in the Neobatrachia support the monophyly of this group; similarly, the tandem duplication of the tRNAMet genes only found in the Dicroglossidae support the monophyly of this family