应用微卫星分型进行小熊猫亲子鉴定

小熊猫栖息于喜马拉雅与横断山脉,是亚洲濒危物种。中国的小熊猫谱系已登记圈养小熊猫个体９６８ 只,现有存活个体３５５ 只（２０１３ 年）,然而,现有谱系中存在部分信息不明确的问题。为此,我们选择１９ 对小熊猫特异性引物对４１ 只圈养小熊猫进行微卫星扩增,并分析其亲缘关系。将获取的亲子鉴定结果,结合种群原始记录信息,利用Pedigraph 软件构建福州圈养小熊猫的种群遗传谱系图。结果表明：在母子关系确实的情况下,１９个基因座位的联合父权排除概率为０. ９９９９９９６８;利用６ 个已知双亲的后代检验１９ 个基因座位的可信度,后代与双亲的基因型完全符合孟德尔遗传定律,表明１９个基因座位能有效确认小熊猫的亲权关系。应用１９ 个微卫星标记成功地为１５ 个后代找到了生物学父亲。尽管一些雌性个体在繁殖期有多重交配的行为,亲子鉴定的结果显示同一窝小熊猫均来自单一父权。基因型比对结果表明７ 对双胞胎均属于异卵双生子。福州种群的后代中除＃９２０和＃９２１ 外,其余均源于＃４８７ 或＃８９８ 两只雄性,表明不同个体参与繁殖的机会不均等。因此,利用现代繁殖技术,加强濒危野动物种群管理,科学制定繁殖计划具有积极的现实意义。     

Effects of mycorrhizae and chitin-hydrolysing microbes on Vicia faba

The effect of Streptomyces albovinaceus (S-22) and Bacillus sp. (B1) on the growth response, nodulation, nutrition and nitrogenase activities of faba bean (Vicia faba) varieties infected with Glomus mosseae under pot conditions in sterile soil amended with chitin was studied. The growth, nodulation, nutrients content and nitrogenase activity of mycorrhiza-treated plants of Giza-667 were significantly increased compared to untreated ones. Such increases were related to the increase in mycorrhizal root infection. Amendment of soil with chitin alone reduced the growth, nodulation, total nitrogen contents and nitrogenase activities of mycorrhiza-treated faba bean plants (Giza-667) compared to untreated plants. Inoculation of plants with S. albovinaceus or Bacillus sp. significantly increased the level of mycorrhizal roots infection, but addition of chitin to the soil in combination with Bacillus sp. reduced the mycorrhizal infection of faba bean roots. Highest phosphorus contents of faba bean Giza-667 were recorded after G. mosseae inoculation in the presence of all treatments. Similar results were observed for the other varieties. The microbial populations were significantly increased in rhizospheres amended with chitin. Such increases were not in response to the mycorrhizal inoculation. Generally, the microflora of faba bean rhizospheres was increased after treatment with G. mosseae in the absence of chitin amendment alone compared with non-mycorrhizal rhizospheres.     

Homogeneous oxidative coupling catalysts: stoichiometry and characterization of the first stable oxotetranuclear solids [(Pip)nCuX]4O2 (n=1 or 2, Pip=piperidine, X=Cl, Br, I)

Copper(I) halides react quantitatively with piperidine (Pip) in dioxygen-free methylene chloride or nitrobenzene to form tetranuclear copper(I) complexes [(Pip)_nCuX]₄; n=1 or 2, X=Cl, Br or I. These complexes are very soluble and completely reduce dioxygen to dioxo bridging ligand, with stoichiometry, Δ[Cu(I)]/Δ[O₂]=4.0. The stable oxo solids [(Pip)_nCuX]₄O₂ mimic tyrosinase copper protein. They act as a homogeneous oxidative coupling catalysts for phenols. Electronic transition spectra in the near infrared with high molecular absorptivity are diagnostic for tetranuclear “Cu₄X₄” core structure. The electronic transitions are more likely due to charge transfer between a minimum of three halo ligands and copper(II) center. The room temperature EPR spectra of [(Pip)_nCuX]₄O₂ in methylene chloride are isotropic with four hyperfine lines. The room temperature solid-state EPR spectra of [PipCuX]₄O₂ show an axial spectra with d_x²−y² ground state, suggesting square pyramidal arrangement of the five coordinated ligands around copper(II) centers. Cyclic voltammetry measurements show that they are more likely irreversible in character and show slight quasi-reversability when X=Br or I. Constant potential electrolysis indicate that the number of electrons consumed are equal to four electrons which will be due to the reduction of four copper(II) to copper(I).     

Synthesis and biological evaluation of 2-styrylquinolines as antitumour agents and EGFR kinase inhibitors: molecular docking study

A new series of 4,6-disubstituted 2-(4-(dimethylamino)styryl)quinoline 4a,b–9a,b was synthesized by the reaction of 2-(4-(dimethylamino)styryl)-6-substituted quinoline-4-carboxylic acids 3a,b with thiosemicarbazide, p-hydroxybenzaldehyde, ethylcyanoacetate, and 2,4-pentandione. In addition, the antitumour activity of all synthesized compounds 3a,b–9a,b was studied via MTT assay against two cancer cell lines (HepG2 and HCT116). Furthermore, epidermal growth factor receptor (EGFR) inhibition, using the most potent antitumour compounds, 3a, 3b, 4a, 4b, and 8a, was evaluated. The interpretation of the results showed clearly that the derivatives 3a, 4a, and 4b exhibited the highest antitumour activities against the tested cell lines HepG2 and HCT116 with IC₅₀ range of 7.7–14.2?µg/ml, in comparison with the reference drugs 5-fluorouracil (IC₅₀?=?7.9 and 5.3?µg/ml, respectively) and afatinib (IC₅₀?=?5.4 and 11.4?µg/ml, respectively). In vitro EGFR screening showed that compounds 3a, 3b, 4a, 4b, and 8a exhibited moderate inhibition towards EGFR with IC₅₀ values at micromolar levels (IC₅₀ range of 16.01–1.11?µM) compared with the reference drugs sorafenib (IC₅₀ =?1.14?µM) and erlotinib (IC₅₀ =?0.1?µM). Molecular docking was performed to study the mode of interaction of compounds 3a and 4b with EGFR kinase.     

Use of mycorrhizal fungi and phosphorus fertilization to improve the yield of onion (Allium cepa L.) plant

Improving the economical yield of commonly cultivated crops is one of the most pressing social and scientific issues in modern agriculture. This paper was conducted to investigate the bio-efficacy of arbuscular mycorrhizal fungi (AMF) in improving phosphorous (P) utilization and increasing the yield of onion plant grown in sandy soil under a drip irrigation system. The obtained results showed that AMF inoculation of onion and application of 120 kg P fertilizer ha⁻¹ significantly increased the fresh and dry weights, chlorophyll content of onion as well as P concentration in the root, shoot, and bulb during two growing seasons. Moreover, AMF increased the bioavailability of P in the rhizosphere and significantly enhanced the N-utilization by the inoculated plant. The economic yield of the onion plant inoculated by AMF and fertilized by different doses of P fertilizer was much higher than that obtained by the control (without AMF). These findings indicated that inoculating the onion plant in the field with AMF could be very effective in increasing the yield of the onion plant. Additionally, this study suggests AMF as a low-cost and promising candidate for the sustainable production of the onion crop using reclaimed sandy soils and a drip irrigation system.Keyword: Onion, Mycorrhizal Fungi, Economic yield, P fertilization, N utilization, ¹⁵N     

Degradation of phenylacetate by Acinetobacter spp.: evidence for the phenylacetyl-coenzyme A pathway

The aerobic degradation of phenylacetate (PA) by many bacteria has recently been shown to proceed via an unprecedented catabolic route. A typical feature of this pathway is the transformation of PA to phenylacetyl-coenzyme A (PACoA). However, the aerobic degradation of PA by Acinetobacter spp. is not sufficiently understood. To gain insight into the catabolism of PA by Acinetobacter spp., we isolated several PA-degrading Acinetobacter spp. from a wastewater treatment plant in Germany using enrichment cultures with PA as a sole carbon source. We also conducted in vitro PA transformation assays based on the detection of PACoA. The identification of the isolated bacteria was based on partial 16S rDNA sequences. Phylogenetic analysis revealed that the isolated strains are members of the Acinetobacter group and could be regarded as strains of Acinetobacter spp. The soluble protein fraction obtained from cells cultured on PA-containing medium transformed PA to several intermediates, as detected by thin layer chromatography and autoradiography. The formation of one intermediate was CoA dependent and comigrated with a sample of PACoA, the earliest characteristic intermediate of the PA catabolic pathway, suggesting that the isolated PA-degrading Acinetobacter spp. utilize the recently elucidated PA catabolic pathway. A database search revealed that many Acinetobacter spp. harbor PA catabolic genes analogous to the paa gene cluster of Escherichia coli K-12.     

Spectral properties and inclusion of a hetero-chalcone analogue in organized media of micellar solutions and beta-cyclodextrin

The absorption and fluorescence emission spectral properties of 3-(4'-dimethylaminophenyl)-1-(2-thienyl)prop-2-en-1-one, abbreviated as DMATP, have been investigated in organized media of aqueous micellar and beta-cyclodextrin (CD) solutions. While the absorption spectra are less sensitive to the nature of the added surfactant or CD, the characteristics of the intramolecular charge transfer (ICT) fluorescence are highly sensitive to the properties of the medium. The ICT maximum is strongly blue-shifted with a great enhancement in the fluorescence quantum yield on adding micellar or CD solutions. This indicates the solubilization of DMATP in the micellar core and formation of an inclusion complex with beta-CD. The critical micelle concentration (CMC) as well as the polarity of the micellar core of SDS, CTAB and TX-100 have been determined. The CMC values are in good agreement with the reported values while the polarity is lower indicating that DMATP molecules are incorporated in the micellar core not at the micellar interface. The inclusion constants of binding of DMATP in micellar or CD have been also determined. The thermodynamic parameters of formation of DMATP:CD inclusion complex have been calculated from the temperature dependence of the fluorescence spectra of the formed complex. The negative enthalpy and free energy of formation indicate that the inclusion process is energetically favorable. The highly negative value of formation entropy (DeltaS = -162.3 J mol(-1) K(-1)) reflects the high restrictions imposed on the movement of both the host and included guest molecules which is consistent with the increase of the fluorescence yield and blue shift of the fluorescence maximum.     

Synthesis and Antimicrobial Activity of Some 2-Pyridone Nucleosides Containing a Sulfonamide Moiety

Glycosylation of 2-pyridonesulfonamide 1a,b with glycosyl/galactosyl bromide gave the corresponding glycosides 2a,b, 3a,b, 6a,b, and 7a,b, respectively. Deacetylation of the resulting glycosides gave the corresponding glycosides 4a,b, 5a,b, 8a,b, and 9a,b, respectively, in good yields. Furthermore, reaction of 2-pyridonesulfonamide 1b with lactosyl bromide gave a mixture the corresponding N, O-lactosides 10 and 11, which were deacetylated to give the corresponding glycosides 12 and 13, respectively. The structures of the new synthesized compounds were characterized by using IR, ¹H, ¹³C NMR spectra, and microanalysis. Selected members of these compounds were screened for antimicrobial activity.     

Cuticular and non-cuticular substrate influence on expression of cuticle-degrading enzymes from conidia of an entomopathogenic fungus,Nomuraea rileyi

Larval cuticle fromTrichoplusia ni, Helicoverpa (=Heliothis)zea, andHeliothis virescens and a cellulose substrate were used to quantify release of proteolytic, chitinolytic, and lipolytic enzymes by germinating conidia of the entomopathogenic fungus,Nomuraea rileyi. There was no significant difference in conidial viability incubated withT. ni, H. zea or cellulose substrates. Conidial viability onH. virescens cuticle, however, was significantly lower (ca. 19–25%) than the other three substrates. The presence of cuticle substrates, especially cuticle ofT. ni, stimulated germination. The nature of the substrate influenced both the time and quantity of the enzymes expressed. Specific proteases (aminopeptidase, chymoelastase, trypsin) generally were expressed earlier and/or in greater quantities on cuticular than on the cellulose substrate. Although both chitinolytic enzymes (endochitinase, N-acetylglucosaminidase) were detected on all three cuticular substrates, their activity was substantially lower than that of the proteolytic enzymes. Lipase activity was only minimally present. Early concurrent release of both proteases and chitinases suggested that both may be important in the penetration of the larval integument by germinating conidia ofN. rileyi. Expression of proteases and chitinases, especially aminopeptidase and endochitinase was probably a specific response to cuticle, because little or no activity was expressed on the non-host, cellulose substrate.This article reports the results of research only. Mention of a proprietary product in this paper does not constitute a recommendation for use by the US Department of Agriculture.     

Review of entomopathogenic fungi and nematodes as biological control agents of tephritid fruit flies: current status and a future vision

Several fruit fly species (Diptera: Tephritidae) are invasive pests that damage the quality of fruits in horticultural crops and cause significant value losses worldwide. Management of fruit flies mainly depends on conventional insecticides. Unfortunately, the application of synthetic insecticides has caused environmental pollution, risks for humans and animals, and development of resistance. Furthermore, controlling fruit flies by applying synthetic insecticides is challenging because fruit containing third instars often fall from the tree – subsequently the larvae leave the decaying fruits and pupate in the soil. Consequently, both larvae and pupae are protected from surface-applied insecticides in fruits and soil. So, there is a pressing need for more eco-friendly and selective control measures with new modes of action. Among such measures are entomopathogenic fungi (EPFs) and nematodes (EPNs). I gathered knowledge on past and present research about EPFs and EPNs as biocontrol agents against fruit flies to investigate approaches that may improve their capacities. I also highlighted several recommendations that may help future field studies on the suppression of fruit fly populations by EPFs and EPNs.