农杆菌介导海洋草酸青霉转化体系及聚酮合酶Pks生物学功能*

为研究南海柳珊瑚共附生草酸青霉SCSGAF0023的聚酮合酶(PKS)生物学功能,采用农杆菌介导法构建草酸青霉SCSGAF0023的Pks敲除株ΔPks,比较野生菌株及ΔPks的生长发育及环境适应性差异。以草酸青霉SCSGAF0023分生孢子为受体,p0380-hygB为双元载体,成功实现草酸青霉SCSGAF0023的遗传转化。结果表明:农杆菌浓度为OD₆₀₀=0.5,在200μmol/L 乙酰丁香酮(AS)诱导下与10⁷个/ml草酸青霉SCSGAF0023孢子于25℃共孵育时转化效率最高。基于上述转化体系,成功获得Pks敲除株ΔPks,并首次证实Pks正向调控草酸青霉SCSGAF0023产孢,但不影响其对环境的适应性。这为进一步系统研究真菌PKSs及聚酮化合物对真菌生长发育与环境适应性的影响提供素材。     

IGF-1 concentration patterns and their relationship with follicle development after weaning in young sows fed different pre-mating diets

Piglet birth weight and within-litter birth weight variation are important for piglet survival and growth. Pre-mating diets may improve IGF-1 and follicle development during the weaning-to-oestrus interval (WEI) and subsequent piglet birth weight. The objective of this study was to modulate IGF-1 concentration during late lactation and the WEI of young sows by using specific pre-mating diets supplemented with microfibrillated cellulose (MF), l-carnitine (LC) or l-arginine (AR). A further objective was to investigate the relationship between IGF-1 and subsequent follicle development and oestrus and ovulation characteristics. In total, 56 first-parity and 20 second-parity sows in three consecutive batches were used for this experiment. Sows received daily either wheat (CON) or wheat plus MF, LC or AR at one of two supplementation levels (low and high) during last week of lactation and WEI. From weaning onwards, follicle and corpus luteum (CL) diameters were repeatedly measured with ultrasound. Blood samples were collected during the WEI for IGF-1 and on day 21 of pregnancy for progesterone analyses, respectively. Insulin-like growth factor-1 concentration, follicle diameter, oestrus and ovulation characteristics and CL diameter were not affected by pre-mating diets. Low IGF-1 class (≤156 ng/ml, N = 22) sows had smaller follicles at weaning (3.5 v. 3.8 mm, P < 0.05) and a longer weaning-to-ovulation interval (147.2 v. 129.8 h, P < 0.05) than high IGF-1 class sows. In first-parity sows, high loin muscle depth (LM) loss sows (≥8%, N = 28) had lower IGF-1 concentrations at weaning (167 v. 214 ng/ml, P < 0.05) compared to low LM loss sows (<8%, N = 28). However, after weaning, IGF-1 concentrations increased and did not differ between high LM loss and low LM loss sows. In conclusion, the different supplemented compounds in pre-mating diets did not improve IGF-1 concentrations around weaning in young sows. Furthermore, high body condition loss caused lower IGF-1 concentrations at weaning, but these levels rapidly recovered after weaning and were related to follicle development and the interval from weaning to ovulation.     

Analysis of Alkylphenol ethoxylates (APEOs) from tannery sediments using LC–MS and their environmental risks

Alkyl phenol polyethoxylates (APEOs) are a major group of high production volume chemicals, extensively used as nonionic surfactants in industrial, agricultural and domestic sectors. These surfactants and its respective metabolites are found to be persistent and toxic. Mainly, they act as endocrine disruptors by mimicking the natural hormones. India being a developing country, witnesses pollution due to various industries including tannery. In the present study, sediment samples were collected from the Vellore district of Tamil Nadu, India to investigate the occurrence of APEOs. The sediments were extracted by ultrasonication and analyzed in a liquid chromatography mass spectrometer. Octyl phenol ethoxylates (OPEOs) were not observed in any of the samples. Nonyl phenol ethoxylates (NPEOs) were detected in the range of ND – 36 mg/kg with 85 % detection frequency. The occurrence of NPEOs in sediment indicates its wide usage in tannery and its partitioning behavior in environment. The levels of NPEOs in the study were found to be unsafe according to the sediment guidelines of various studies. Since NPEOs were observed in sediment samples, possibilities of occurrence of their monomers and metabolites cannot be ruled out. Therefore, further studies are warranted for understanding the levels of monomers and metabolites in order to ascertain the environmental risks more appropriately.     

Does operational sex ratio influence relative strength of purging selection in males versus females?

According to theory, sexual selection in males may efficiently purge mutation load of sexual populations, reducing or fully compensating ‘the cost of males’. For this to occur, mutations not only need to be deleterious to both sexes, they also must affect males more than females. A frequently overlooked problem is that relative strength of selection on males versus females may vary between environments, with social conditions being particularly likely to affect selection in males and females differently. Here, we induced mutations in red flour beetles (Tribolium castaneum) and tested their effect in both sexes under three different operational sex ratios (1:2, 1:1 and 2:1). Induced mutations decreased fitness of both males and females, but their effect was not stronger in males. Surprisingly, operational sex ratio did not affect selection against deleterious mutations nor its relative strength in the sexes. Thus, our results show no support for the role of sexual selection in the evolutionary maintenance of sex.     

谷氨酸棒杆菌碱基编辑的条件优化

近年来,基于CRISPR/Cas9的碱基编辑技术因其具有不产生DNA双链断裂、无需外源DNA模板、不依赖宿主同源重组修复的优势,已经逐渐发展成为一种强大的基因组编辑工具,在动物、植物、酵母和细菌中得到了开发和应用。研究团队前期已在重要的工业模式菌株谷氨酸棒杆菌中开发了一种多元自动化的碱基编辑技术MACBETH,为进一步优化该方法,提高碱基编辑技术在谷氨酸棒杆菌中的应用效率,本研究首先在谷氨酸棒杆菌中构建了基于绿色荧光蛋白(GFP)的检测系统:将GFP基因的起始密码子ATG人工突变为ACG,GFP无法正常表达,当该密码子的C经编辑后恢复为T,即实现GFP蛋白的复活,结合流式细胞仪分析技术,可快速衡量编辑效率。然后,构建针对靶标位点的碱基编辑工具,经测试,该位点可成功被编辑,在初始编辑条件下碱基编辑效率为(13.11±0.21)%。在此基础上,通过对不同培养基类型、诱导初始OD600、诱导时间、诱导物浓度进行优化,确定最优编辑条件是:培养基为CGXII,初始OD600为0.05,诱导时间为20 h,IPTG浓度为0.01 mmol/L。经过优化,编辑效率达到(30.35±0.75)%,较初始条件提高了1.3倍。最后,选取原编辑条件下编辑效率较低的位点,进行了优化后编辑条件下的编辑效率评估,结果显示,不同的位点在最优编辑条件下的编辑效率提高了1.7–2.5倍,进一步证实该优化条件的有效性及通用性。研究结果为碱基编辑技术在谷氨酸棒杆菌中更好的应用提供了重要的参考价值。     

Selenium nanovirus and its cytotoxicity in selenite-exposed higher living organisms

Selenium (Se) is an essential micronutrient in living organisms, having a narrow margin between essential and potentially toxic intake/exposure. Thus, the biochemistry of Se in living organisms must be studied in-depth to determine the underlying mechanism of Se cytotoxicity. In this study, we report the emergence of selenium nanovirus (SeNVs) in selenite-exposed fish (freshwater and saltwater) and plants (dryland) and its toxicity in them. SeNVs were found in both the abdomen and tail of Oryzias melastigma and saltwater Rhodeus ocellatus, which led to their death. The occurrence of the intracellular assembly of SeNVs was observed in the roots and leaves of corn Zea mays, but not in those of Limnobium laevigatum. SeNVs led to the death of Z. mays but caused chronic toxicity in L. laevigatum. SeNVs should be a system or structure that dissipates the intracellular redox gradients of the host cells, with simple information consisting Se–O, Se–N, or Se–S bond, that would ensure elemental Se ligand binding with nearly specific biomolecules in host cells, thereby maintaining their composition and stabilizing their structure. The multiple toxic effects of Se, therefore, could be the consequence of increase of entropy in the host cells caused by the intracellular assembly of SeNVs. This study may provide an insight into the underlying mechanism of Se in environmental toxicology and its applications in human health.     

The poor health of deep-water species in the context of fishing activity and a warming climate: will populations of Molva species rebuild or collapse?

Many deep-water fish populations, being K-selected species, have little resilience to overexploitation and may be at serious risk of depletion as a consequence. Sea warming represents an additional threat. In this study, the condition, or health, of several populations of common ling (Molva molva), blue ling (Molva dypterygia) and Mediterranean or Spanish ling (Molva macrophthalma) inhabiting different areas in the North Atlantic and the Mediterranean was evaluated, to shed light on the challenges these deep-water species are facing in the context of fishing activity and a warming climate. The data on the condition of Molva populations which are analysed here have been complemented with data on abundance and, for the southernmost species (Mediterranean ling), with two other health indicators (parasitism and hepato-somatic index). Despite some exceptions (e.g., common ling in Icelandic waters), this study shows that the condition of many populations of Molva species in the northeastern Atlantic and the Mediterranean Sea has worsened, a trend which, in recent decades, has usually been found to be accompanied by a decline in their abundance. In addition, the poor health status of most populations of common ling, blue ling and Mediterranean ling considered in this analysis points to a lower sustainability of these populations in the future. Overall, the health status and abundance of Molva populations in the northeastern Atlantic and the Mediterranean suggest that only some populations located in the North Atlantic may be able to rebuild, whereas the populations in southern North Atlantic and the Mediterranean, which are probably most at risk from sea warming, are facing serious difficulties in doing so. In the context of fisheries and global warming, this study's results strongly indicate that management bodies need to consider the health status of many of the populations of Molva species, particularly in southern European waters, before implementing their decisions.     

Effects of Radio-Collars are not Contingent on Socioecological Conditions in Degus

Species-specific research on free-ranging mammals reveals a diversity of effects of radio-collars on behavior, body condition, and fitness. Although these studies indicate rather limited direct effects, radio-collars may cause effects influenced by socio-ecological conditions. Using a 7-year study on a natural population of group-living degus (Octodon degus), we tested the hypothesis that ecological (food availability, burrow density) and social (group size, group male-to-female ratio) conditions modulate effects of radio-collars on body condition (e.g., body mass, ecto- and endoparasite loads, fecal cortisol metabolites) and direct fitness (litter size, adult survival). We determined the effect of radio-collar use on degus by contrasting the presence or absence of radio-collars, quantifying the effects of the number of days carrying a radio-collar, and the relative mass of radio-collars worn by degus in central Chile between 2009 and 2015. Radio-collar use was not associated with direct effects on litter size, adult survival, or with body mass and fecal cortisol metabolites but was linked to low ecto- and endoparasite loads. These seemingly positive effects may reflect decreased mobility, or a research bias for radio-collaring larger, healthier individuals. There was no evidence that ecological and social conditions modulated radio-collar effects on degu body condition and direct fitness. These findings are consistent with evidence from other mammal studies that reported no appreciable detrimental direct or indirect effects of radio-collars. © 2021 The Wildlife Society.     

西南地区壳斗科物种丰富度和特有性分布格局模拟及其环境解释

如何准确地模拟物种宏观丰富度格局和特有性中心是生物多样性保护工作的重点,也是生物地理学的热点话题。西南地区是我国壳斗科植物最丰富的地区之一,但物种多样性格局及环境驱动机制尚不清楚。本研究基于西南地区161种壳斗科植物7258个分布点位数据,利用点格局法和物种分布模型两种方式构建了物种丰富度、加权特有性指数和校正加权特有性指数的分布格局,并采用空间自回归模型(SAR)分析上述3个多样性指数与环境因子间的关系。总体上看,物种分布模型模拟的3个指数在空间上比点格局法更为连续,但数值高低分布情况具有相似性: 两种方式模拟的物种丰富度高值区主要分布在滇南边缘、桂北部和桂西南部地区(62～89种);加权特有性指数最大值集中在滇南和桂西地区(1.77～5.02);藏东南、秦岭-大巴山、桂西南部和滇东南地区具有最高的校正加权特有性指数(0.07～0.17)。SAR模型结果显示:最干月降雨量、温度季节性变化标准差、海拔变幅和土壤有机碳含量对物种丰富度的影响均显著,最干月降雨量、温度季节性变化标准差、潜在蒸散量和海拔变幅对加权特有性有着显著影响,温度季节性变化标准差、最干月降雨量、历史温度变化、增强型植被指数变异系数和海拔变幅对校正加权特有性的影响显著;SAR模型对物种丰富度、特有性指数和加权特有性指数的拟合效果(R²=0.857、0.733、0.593)分别优于普通线性模型(R²=0.689、0.425、0.422)。综上,水分可获得性、气候季节性、生境异质性、历史气候变化和土壤状况是制约西南地区壳斗科丰富度和特有性分布的最重要因素。滇南、滇东南、桂西南、桂西、秦岭-大巴山以及藏东南地区是壳斗科物种丰富度中心或特有性中心,应受到重点关注和保护。