The genetic structure of the A mating-type locus of Lentinula edodes

The Shiitake mushroom, Lentinula edodes (Berk.) Pegler is a tetrapolar basidiomycete with two unlinked mating-type loci, commonly called the A and B loci. Identifying the mating-types in shiitake is important for enhancing the breeding and cultivation of this economically-important edible mushroom. Here, we identified the A mating-type locus from the first draft genome sequence of L. edodes and characterized multiple alleles from different monokaryotic strains. Two intron-length polymorphism markers were developed to facilitate rapid molecular determination of A mating-type. L. edodes sequences were compared with those of known tetrapolar and bipolar basidiomycete species. The A mating-type genes are conserved at the homeodomain region across the order Agaricales. However, we observed unique genomic organization of the locus in L. edodes which exhibits atypical gene order and multiple repetitive elements around its A locus. To our knowledge, this is the first known exception among Homobasidiomycetes, in which the mitochondrial intermediate peptidase (mip) gene is not closely linked to A locus.     

Diterpene acids from Helianthus species and their microbiological conversion by Gibberella fujikuroi, mutant B1-41a

The diterpene acid content in 10 species of Helianthus has been investigated. Ent-12,16-cyclokauranoic acid, isolated from H. annuus, is converted into a series of 12,16-cyclogibberellins by cultures of Gibberella fujikuroi, mutant B1-41a, and 12,16-cyclogibberellins A₉, and A₁₂ have been isolated. Ent-12β-acetoxykaurenoic acid and ent-13(S)-angeloxyatisenoic acid have been isolated from H. decapetalus; the metabolism of ent-13(S)-hydroxyatisenoic acid and atisenoic acid by B1-41a is also described.     

Phenolic constituents of Semecarpus anacardium

GLC-MS analysis of methylated bhilawanol from S. anacardium nuts and its oxidation product, the methyl ester of an aromatic carboxylic acid, conclusively proved that it contains more than seven closely related compounds. Two of them are major components which were isolated and shown to be 1-pentadec-Δ^5′-enyl-2,3-dimethoxybenzene (I) and 1-pent biflavanoids A, B and C have been also isolated from defatted nuts of S. anacardium. The first of these has been characterized as its methyl ethers. A₁ and A₂, for which biflavanone structures (VI) and (VII) are suggested on the basis of chemical and spectral evidence. The biflavanones B and C have been also characterized as their methyl ethers. Suggested structures are O-methyl derivatives of a IB-3′, IIA-8-binaringenin (XIV) for the former and IB-3′, IIA-8-biliquiritigenin (XV) for the latter.     

Mutation and expression analysis of the IDH1, IDH2, DNMT3A, and MYD88 genes in colorectal cancer

Colorectal cancer (CRC) is one of the leading causes of death around the world. Its genetic mechanism was intensively investigated in the past decades with findings of a number of canonical oncogenes and tumor-suppressor genes such as APC, KRAS, and TP53. Recent genome-wide association and sequencing studies have identified a series of promising oncogenes including IDH1, IDH2, DNMT3A, and MYD88 in hematologic malignancies. However, whether these genes are involved in CRC remains unknown. In this study, we screened the hotspot mutations of these four genes in 305 CRC samples from Han Chinese by direct sequencing. mRNA expression levels of these genes were quantified by quantitative real-time PCR (RT-qPCR) in paired cancerous and paracancerous tissues. Association analyses between mRNA expression levels and different cancerous stages were performed. Except for one patient harboring IDH1 mutation p.I99M, we identified no previously reported hotspot mutations in colorectal cancer tissues. mRNA expression levels of IDH1, DNMT3A, and MYD88, but not IDH2, were significantly decreased in the cancerous tissues comparing with the paired paracancerous normal tissues. Taken together, the hotspot mutations of IDH1, IDH2, DNMT3A, and MYD88 gene were absent in CRC. Aberrant mRNA expression of IDH1, DNMT3A, and MYD88 gene might be actively involved in the development of CRC.     

Why the aphid Aphis spiraecola is more abundant on clementine tree than Aphis gossypii?

Aphis spiraecola and Aphis gossypii cause harmful damages on clementine tree orchards. Weekly surveys measured the abundance of aphids (larvae, winged and wingless adults) as well as of auxiliary insects and parameters of energy metabolism. Correlatively, soluble carbohydrates, total free amino acids, free proline and condensed tannins were quantified in control and infested leaves. Both aphid species showed parallel temporal variations, but A. spiraecola was consistently more abundant regardless of the stage. Amino acids had a positive effect on both aphid species abundance, but neither condensed tannins nor auxiliary insects seemed to modulate aphid populations. Interestingly, the leaf carbohydrate content was positively correlated with the abundance of A. spiraecola, but not with that of A. gossypii. Moreover, A. gossypii's abundance was significantly down-regulated by high proline concentrations. Thus, the higher abundance of A. spiraecola could be explained by a better tolerance to high proline contents and a better conversion of foliar energy metabolites.     

Toxic tetranortriterpenes of the fruit of Melia azedarach

Four new tetranortriterpenes, meliatoxins A₁, A₂, B₁ and B₂ have been isolated and identified as toxic constituents of the fruit of Melia azedarach L. var. australasica. Toxicity and pathological results confirm that the meliatoxins are responsible for most but not all of the symptoms resulting from the ingestion of whole fruit.     

1914G variant of BCHE gene associated with enzyme activity,obesity and triglyceride levels

Polymorphisms of butyrylcholinesterase (BChE) have been reported to be associated to weight, BMI variance and hypertriglyceridemia in adults and adolescents. The aim of the present study was to investigate the association of −116A (SNP: G/A; rs1126680) and 1914G (SNP: A/G; rs3495) variants of BCHE gene with anthropometric and biochemical variables associated with obesity in population sample of 115 individuals, from Southern Brazil. Participants were grouped in two categories: obese (BMI ≥ 30) and non-obese (BMI < 30). The 1914G allele showed significantly higher frequency in the obese group, and carriers of 1914G allele showed lower mean BChE activity when compared to 1914A carriers (p = 0.006). Higher means of BMI (p = 0.02) and triglyceride (TG; p = 0.01) were found in 1914G carriers (BMI = 27.57kg/m²; TG = 150.8 mg/dL) when compared to 1914A homozygotes (BMI = 25.55 kg/m²; TG = 107.9 mg/dL). Carriers of the −116A allele showed lower mean BChE activity than usual homozygotes, and the −116A variant was found in cis with 1914G (p < 0.0001; D′ = 1). The region of BCHE gene that contains the 1914G mutation site is target of microRNAs (miRs) and the response of BChE to glucocorticoids is especially influenced by these miRs. Therefore, it is possible that the 1914G allele can be interfering in gluconeogenesis, hyperglycemia, lipolysis and body fat distribution. This lower activity may cause an imbalance in lipid metabolism, which may lead to an increased predisposition to obesity and to a lower ability to maintain metabolic homeostasis.     

New chromenochalcones from Flemingia

2-Methyl-2-(4′-methylpent-3′-enyl)-5,8-dihydroxy-6-(4-hydroxycinnamoyl)-chromene (flemingin-D) has been isolated from inflorescences of Flemingia congesta, together with the known flemingin-C and two other chalcones. These (flemingin-E and -F) are allylic alcohols which differ from flemingin-D by an additional OH group in the side chain. Photooxidation of model 2-methyl-2-(4′-methylpent-3′-enyl)-chromenes gave pairs of allylic alcohols with the same structural feature. The known flemingin-A, -B, -C and homoflemingins have been isolated from F. grahamiana, and another new chalcone from F. bracteata.     

Peroxidases of Solanum melongena leaves

Three major peroxidases, designated as A, B₂ and B₂ from Solanum melongena leaves have been reported. Peroxidases-A, -B₂ and -B₂ were considered to be true peroxidases on the basis of k₁:k₄ ratio. The pH optima for the three enzymes were found to be 7·0, 6·0 and 6.0 respectively. These peroxidases differ in their k₁:k₄ ratio, in the effect of pH on this ratio and in the uric acid/guaiacol and o-dianisidine/guaiacol activity ratio.     

Association of IL-17A and IL-17F single nucleotide polymorphisms with susceptibility to osteoarthritis in a Korean population

The damage incurred in osteoarthritis (OA) is mediated by a variety of cytokines, growth factors and inflammatory mediators. The importance of the interleukin-17 (IL-17) family in inflammatory and autoimmune disease is becoming increasingly apparent. Microsatellite association mapping reveals a primary osteoarthritis susceptibility locus on chromosome 6p12.3-q13. IL-17A and IL-17F genes that resided on chromosome 6p12.3-q13 are believed to play an important role in the primary OA susceptibility. We investigated the allele and genotype of IL-17A G-197A and IL-17F T7488C in 302 OA patients and 300 healthy subjects as controls. We employed a PCR-SSCP assay to identify the genotypes IL-17A G-197A and IL-17F T7488C. For IL-17A G-197A, there were significant differences in frequencies of genotype and allele of IL-17A G-197A between OA patients and controls (both p < 0.0001). For IL-17F T7488C, there were no significant differences in the allele frequency and genotype distribution for IL-17F T7488C between OA patients and controls (p = 0.938 and p = 0.1735, respectively). In conclusion, current study showed that polymorphism of IL-17A G-197A may be closely associated with susceptibility to the development of OA in the Korean population. However, there was no relationship between IL-17F T7488C polymorphism and OA susceptibility.     

Insecticidal activity and chemical composition of the essential oil of Artemisia vestita from China against Sitophilus zeamais

In our screening program for new agrochemicals from local wild plants, essential oil of Artemisia vestita Wall (Asteraceae) was found to possess strong insecticidal activity against maize weevil, Sitophilus zeamais Motsch. Essential oil of aerial parts of A. vestita was obtained from hydrodistillation and was investigated by GC and GC–MS. The main components of essential oil were grandisol (40.29%), 1,8-cineol (14.88%) and camphor (11.37%). The essential oil of A. vestita possessed strong fumigant toxicity against S. zeamais adults with a LC₅₀ value of 13.42 mg/L air. The essential oil of A. vestita also showed contact toxicity against S. zeamais adults with a LD₅₀ value of 50.62 mg/adult.     

Depsipeptides from Metarhizium anisopliae

From the culture medium of a strain of Metarhizium anisopliae, 14 depsipeptides have been isolated. Five of them were identified as known destruxins A, B, C, D and desmethyldestruxin B. The structures of the new compounds, named destruxins E, A₁, A₂, B₁, B₂, C₂, D₁, D₂ and E₁,were established mainly from the mass spectral analysis of their corresponding open-chain derivatives.     

Ribosomal Protein Genes RPS10 and RPS26 Are Commonly Mutated in Diamond-Blackfan Anemia

Diamond-Blackfan anemia (DBA), an inherited bone marrow failure syndrome characterized by anemia that usually presents before the first birthday or in early childhood, is associated with birth defects and an increased risk of cancer. Although anemia is the most prominent feature of DBA, the disease is also characterized by growth retardation and congenital malformations, in particular craniofacial, upper limb, heart, and urinary system defects that are present in ∼30%–50% of patients. DBA has been associated with mutations in seven ribosomal protein (RP) genes, RPS19, RPS24, RPS17, RPL35A, RPL5, RPL11, and RPS7, in about 43% of patients. To continue our large-scale screen of RP genes in a DBA population, we sequenced 35 ribosomal protein genes, RPL15, RPL24, RPL29, RPL32, RPL34, RPL9, RPL37, RPS14, RPS23, RPL10A, RPS10, RPS12, RPS18, RPL30, RPS20, RPL12, RPL7A, RPS6, RPL27A, RPLP2, RPS25, RPS3, RPL41, RPL6, RPLP0, RPS26, RPL21, RPL36AL, RPS29, RPL4, RPLP1, RPL13, RPS15A, RPS2, and RPL38, in our DBA patient cohort of 117 probands. We identified three distinct mutations of RPS10 in five probands and nine distinct mutations of RPS26 in 12 probands. Pre-rRNA analysis in lymphoblastoid cells from patients bearing mutations in RPS10 and RPS26 showed elevated levels of 18S-E pre-rRNA. This accumulation is consistent with the phenotype observed in HeLa cells after knockdown of RPS10 or RPS26 expression with siRNAs, which indicates that mutations in the RPS10 and RPS26 genes in DBA patients affect the function of the proteins in rRNA processing.     

八肋游仆虫大核基因组中组蛋白基因 H4A和H4B的克隆及分析

组蛋白作为核小体的基本组分,是染色质的结构和功能必需的。组蛋白的变体和修饰共同参与染色质修饰及基因的表达调控。真核生物细胞中的5种组蛋白在进化中高度保守,然而纤毛虫的组蛋白H4与其他真核生物相比有较大的差异。本实验应用PCR技术从八肋游仆虫(Euplotes octocarinatus)中获得了2种组蛋白H4基因,分别为H4A和H4B,GenBank登录号为:JN715068和JN715069。序列分析表明,H4A基因开放阅读框324 bp,预测编码107个氨基酸,分子量为11.6 ku,等电点为10.99。而H4B基因编码框384 bp,编码127个氨基酸,分子量为14.4 ku,等电点为9.93。Blast结果显示,H4A序列与其他生物中H4的一致性相对较高,达81%～94%,而H4B的一致性为36%～70%。H4A和H4B的一致性仅为44.7%。实时荧光定量PCR表明,H4A的转录本高于H4B。结果提示:在进化过程中八肋游仆虫可能进化出特殊的组蛋白H4基因,不同的组蛋白H4可能发挥不同的功能。     

Sesquiterpenes from Cacalia hastata

From the roots of Cacalia hastata L. subsp. orientalis Kitamura 5 sesquiterpenes were isolated; fukinone (I), dehydrofukinone (II), bakkenolide A (III), cacalohastine (IV) and dehydrocacalohastine (V).     

Spectroscopic studies of the chlorophyll d containing photosystem I from the cyanobacterium, Acaryochloris marina

Absorbance difference spectroscopy and redox titrations have been applied to investigate the properties of photosystem I from the chlorophyll d containing cyanobacterium Acaryochloris marina. At room temperature, the (P740⁺ − P740) and (F_A/B⁻ − F_A/B) absorbance difference spectra were recorded in the range between 300 and 1000 nm while at cryogenic temperatures, (P740⁺A₁⁻ − P740A₁) and (³P740 − P740) absorbance difference spectra have been measured. Spectroscopic and kinetic evidence is presented that the cofactors involved in the electron transfer from the reduced secondary electron acceptor, phylloquinone (A₁⁻), to the terminal electron acceptor and their structural arrangement are virtually identical to those of chlorophyll a containing photosystem I. The oxidation potential of the primary electron donor P740 of photosystem I has been reinvestigated. We find a midpoint potential of 450 ± 10 mV in photosystem I-enriched membrane fractions as well as in thylakoids which is very similar to that found for P700 in chlorophyll a dominated organisms. In addition, the extinction difference coefficient for the oxidation of the primary donor has been determined and a value of 45,000 ± 4000 M^− 1 cm^− 1 at 740 nm was obtained. Based on this value the ratio of P740 to chlorophyll is calculated to be 1:~ 200 chlorophyll d in thylakoid membranes. The consequences of our findings for the energetics in photosystem I of A. marina are discussed as well as the pigment stoichiometry and spectral characteristics of P740.     

Salt Induces Expression of RH3.2A, Encoding an H3.2-type Histone H3 Protein in Rice (Oryza sativa L.)

Histone H3 is one of the four histones, along with H2A, H2B, and H4, which form the eukaryotic nucleosome octamer core. In this study, a new gene RH3.2A encoding an H3.2-type histone H3 protein from rice (Oryza sativa L.) was reported. RH3.2A was cloned through RT-PCR from salt-treated rice seedlings. This gene encoded a protein of 136 amino acid residues that were similar to some plant histone H3 proteins reported previously. However, the cDNA sequence of RH3.2A and other rice H3 genes were different. Alignment of RH3.2A encoding protein with other plant histone H3 proteins revealed that three amino acid residues (32, 88, and 91) were markedly different between H3.1-type and H3.2-type proteins. The mRNA expression analysis of RH3.2A revealed that RH3.2A gene was upregulated by salt stress in rice roots and ABA treatment in seedlings. The potential role of RH3.2A during salt stress was discussed.     

RFLP and AFLP Analysis of Inter- and Intraspecific Variation of Brassica rapa and B. napus Shows that B. rapa Is an Important Genetic Resource for B. napus Improvement

Fingerprinting of 29 accessions of oilseed rape, including seven accessions of Brassica napus, and 22 accessions of B. rapa (B. campestris) from Europe, North America, and China was analyzed using RFLP and AFLP markers. In total, 1 477 polymorphic RFLP bands and 183 polymorphic AFLP bands from 166 enzyme-probe combinations and two pairs of AFLP primers, respectively, were scored for the 29 accessions. On average, RFLP analysis showed that the Arabidopsis EST probe detected more polymorphic bands in Brassica than the random genomic probe performed. More polymorphic RFLP markers were detected with the digestion of EcoR I or BamH I than HindIII. According to the number of bands amplified from each accession, the copy numbers of each gene in the genomes of B. rapa and B. napus were estimated. The average copy numbers in B. rapa of China, B. rapa of Europe, and B. napus, were 3.2, 3.1, and 2.9, respectively. Genetic distance based on the AFLP data was well correlated with that based on the RFLP data (r = 0.72, P<0.001), but 0.39 smaller on average. Genetic diversity analysis showed that Chinese B. rapa was more polymorphic than Chinese B. napus and European materials. Some European B. napus accessions were clustered into European B. rapa, which were distinctly different from Chinese B. napus. The larger variations of Chinese accessions of B. rapa suggest that they are valuable in oilseed rape breeding. Novel strategies to use intersubgenomic heterosis between genome of B. rapa (A^rA^r) and genome of B. napus (AⁿAⁿCⁿCⁿ) were elucidated.