番木瓜果糖-2,6-二磷酸酶CpF2KP基因克隆和蛋白表达纯化

番木瓜是岭南四大名果之一,在我国东南部地区广泛种植,因其具有食用和药用双重价值,因此深受人们的青睐。果糖-6-磷酸,2-激酶/果糖-2,6-二磷酸酯酶(fructose-6-phosphate,2-kinase/fructose-2,6-bisphosphatase,F2KP)是一个独特的双功能酶,具有激酶功能域和酯酶功能域,能催化生物体内糖代谢的重要调节物果糖-2,6-二磷酸(Fru-2,6-P_(2))的合成和降解。为了研究番木瓜中编码该酶的基因CpF2KP的功能,得到目的蛋白尤为重要。本研究从番木瓜基因组中提取到CpF2KP基因的编码序列(coding sequence,CDS)序列,该基因CDS全长2274 bp。将该基因CDS全长扩增之后选用pGEX-4T-1载体进行原核表达。对载体pGEX-4T-1用EcoRⅠ和BamHⅠ进行双酶切,利用基因重组的方式将扩增序列构建到原核表达载体上。经过诱导条件探索,SDS-PAGE结果显示GST-CpF2KP重组蛋白的大小约为110 kDa,诱导CpF2KP蛋白表达的最适条件为:异丙基β-D-硫代半乳糖苷(isopropyl beta-D-thiogalactopyranoside,IPTG)浓度为0.5 mmol/L,温度28℃。对诱导后的CpF2KP蛋白进行纯化,得到了纯化的单一目的蛋白。此外,检测了该基因的组织表达特性发现该基因在种子中表达量最高,在果肉中表达量最低。该研究为进一步深入揭示番木瓜CpF2KP蛋白的功能及研究该基因参与的生物学过程提供了重要基础。     

Antibacterial Activity and Mechanisms of Ethanol Extracts from <i>Scutellaria baicalensis</i> Georgi and <i>Magnolia officinalis</i> against <i>Phytophthora nicotianae</i>

Phytophthora nicotianae causes substantial economic losses in most countries where tobacco is produced. At present, the control of P. nicotianae mainly depends on chemical methods, with considerable environmental and health issues. We investigated the effects of ethanol extracts from Scutellaria baicalensis Georgi (SBG) and Magnolia officinalis (MO). On mycelial growth, sporangium formation, and zoospore release of P. nicotianae. Both extracts inhibited the growth of P. nicotianae, with mycelial growth inhibition rates of 88.92% and 93.92%, respectively, at 40 mg/mL, and EC50 values of 5.39 and 5.74 mg/mL, respectively. The underlying mechanisms were the inhibition of sporangium formation, the reduction of zoospore number, and the destruction of the mycelium structure. At an SBG extract concentration of 16.17 mg/mL, the inhibition rates for sporangia and zoospores were 98.66% and 99.39%, respectively. At an MO extract concentration of 2.87 mg/mL, the production of sporangia and zoospores was completely inhibited. The hyphae treated with the two plant extracts showed different degrees of deformation and damage. Hyphae treated with SBG extract showed adhesion and local swelling, whereas treatment with MO extract resulted in broken hyphae. Mixture of the extracts resulted in a good synergistic effect.     

我国巢鼠属分类和分子系统学分析

本研究对2018至2021年采集的9号巢鼠(Micromys minutus)标本、22号红耳巢鼠(M. erythrotis)标本和19号待厘定的巢鼠属标本,进行形态分类和分子系统学分析,进一步揭示我国巢鼠属的分类和系统分化问题。待厘定的巢鼠属标本形态特征为：标本体背毛黑棕,体腹毛基灰色,毛尖灰白,体侧毛色具明显区分,尾背部毛色黑棕,尾腹部毛色灰棕色;尾长长于头体长的120%;头骨背面观可见颧弓明显弯曲;颅全长[(18.59 ± 0.48)mm]和颅基长[(17.43 ± 0.48 mm)]较长,腭长[(9.35 ± 0.11)mm]较长,脑颅高[(7.43 ± 0.06)mm]较高。待厘定的巢鼠属标本形态特征与巢鼠和红耳巢鼠均存在差异。待厘定巢鼠属标本与巢鼠和红耳巢鼠之间的遗传距离分别为0.115和0.136,接近于巢鼠与红耳巢鼠之间的遗传距离(0.126)。利用Cyt b基因全序列和核基因IRBP1、RAG1和RAG2序列分别构建的巢鼠属系统发生树均以较高的置信度分化成3个进化支,即巢鼠、红耳巢鼠和待厘定的巢鼠属样本的进化支。形态学和分子系统学分析结果均支持待厘定的巢鼠属标本为独立物种分类单元,对应于文献记载的巢鼠川西亚种(M. m. pygmaeus)。根据产地、遗传距离和形态分化,建议将巢鼠川西亚种提升为种,命名为川西巢鼠(M. pygmaeus comb. nov.)。利用Cyt b基因全序列构建的巢鼠系统发生树分化成6个进化谱系：日韩谱系、欧洲谱系、俄罗斯新西伯利亚谱系、中国东北和俄罗斯远东谱系、中国安徽谱系和中国台湾谱系。     

牛蛙冬眠期及其前后的消化道嗜银细胞分布

为研究两栖类在冬眠期及其前后消化道嗜银细胞是否参与冬眠期的消化调节,本文以牛蛙(Rana catesbeiana)为实验对象,采用Grimelius银染法,对冬眠期前(n = 10)、冬眠期(n = 10)和冬眠期后(n = 10)牛蛙消化道嗜银细胞的形态及密度进行比较研究。结果表明,牛蛙消化道各部位均有嗜银细胞分布;牛蛙消化道嗜银细胞形态在冬眠期、冬眠期前及冬眠期后无差异,均为锥体型、梭型和椭圆型;牛蛙消化道各部位具有外分泌功能的锥体型和梭形嗜银细胞密度在3个时期均显著高于具有内分泌功能的椭圆型嗜银细胞密度(P < 0.01);3个时期牛蛙消化道嗜银细胞分布密度高峰均位于空肠处,但低谷有所不同,冬眠期前和冬眠期后牛蛙消化道嗜银细胞的分布密度低谷位于食管,而冬眠期其分布密度低谷位于贲门;3个时期相比,冬眠期前和冬眠期幽门处分布密度差异不显著(P > 0.05),其余部位均有差异,且食管、胃、十二指肠、空肠、回肠和直肠中嗜银细胞分布密度在冬眠期显著高于冬眠期前和冬眠期后(P < 0.05);冬眠期前和冬眠期后消化道嗜银细胞分布密度呈倒“U”型趋势,冬眠期分布密度呈现“~”型趋势。结合相关研究,推测牛蛙嗜银细胞分布密度的改变可能与机体适应不同生理状态及消化功能的调节有关。     

乙醛脱氢酶基因ALDH2对雄性小鼠生育能力的影响

本研究使用乙醛脱氢酶2(ALDH2)基因全敲除的雄性C57BL/6(B6)小鼠（Musmusculus domesticus）,通过分析不同周龄小鼠的睾丸脏器系数、睾丸组织细胞形态、精子运动参数、配种后母鼠产仔数及子代雄雌比等生育指标,探讨ALDH2基因敲除对雄性小鼠生育能力的影响。结果表明,与野生型（WT）小鼠相比,5、7、10周龄ALDH2基因敲除型（KO）雄性小鼠睾丸脏器系数显著降低（P <0.05）;睾丸组织细胞间质变大,精子活率显著降低（P <0.05）;产仔数和雄雌比显著降低（P <0.05）。本研究为揭示乙醛脱氢酶ALDH2基因在雄性小鼠生殖中的作用提供了一定的基础。     

湖南郴州发现岭南两头蛇

2022年8月,于湖南省郴州市永兴县湘阴渡镇（113°01′18′′E,26°11′87′′N,海拔174 m）采集到两头蛇活体标本1号,通过形态测量比较,发现所采集标本与中国广东省分布的岭南两头蛇（Calamariaarcana）形态特征基本一致;后基于线粒体Cytb基因片段进行分子系统发育分析,结果显示该标本与岭南两头蛇形成高支持率单系群（后验概率PP为1.00,自举值BS为100）,所采集标本与岭南两头蛇的遗传距离为1%。综合形态学和分子生物学证据,最终确定该标本为两头蛇科（Calamariidae）两头蛇属的岭南两头蛇,系湖南省蛇类分布新记录种。     

氰氟虫腙对草地贪夜蛾的毒力与田间防效

【目的】本研究旨在明确氰氟虫腙对草地贪夜蛾Spodoptera frugiperda幼虫的毒力水平和田间防治效果,为科学使用氰氟虫腙防治草地贪夜蛾提供参考依据。【方法】采用饲料混毒法在室内测定了氰氟虫腙与4种常用杀虫剂甲维盐、氯虫苯甲酰胺、虱螨脲和茚虫威对草地贪夜蛾3和6龄幼虫的致死中浓度(LC₅₀)及LC₉₀值,以及LC₉₀浓度的这些杀虫剂对3龄幼虫的致死中时(medium lethal time, LT₅₀)值。采用人工喷雾方法测定了玉米田中22%氰氟虫腙悬浮剂(6.6 g/667 m²)、22%氰氟虫腙悬浮剂(17.6 g/667 m²)、5.7%甲维盐水分散剂(1 g/667 m²)和150 g/L茚虫威悬浮剂(2 g/667 m²)对草地贪夜蛾幼虫的防效。【结果】室内生测结果显示,供试的5种杀虫剂中氰氟虫腙对草地贪夜蛾3和6龄幼虫均具有较高的毒力,其LC₅₀值分别为2.64和4.3...     

Regulation of Nitrogen Metabolism,Photosynthetic Activity,and Yield Attributes of Spring Wheat by Nitrogen Fertilizer in the Semi-arid Loess Plateau Region

It is critical for spring wheat (Triticum aestivum L.) production in the semi-arid Loess Plateau to understand the impact of nitrogen (N) fertilizer on changes in N metabolism, photosynthetic parameters, and their relationship with grain yield and quality. The photosynthetic capacity of flag leaves, dry matter accumulation, and N metabolite enzyme activities from anthesis to maturity were studied on a long-term fertilization trial under different N rates [0 kg ha^?1(N1), 52.5 kg ha^?1 (N2), 105 kg ha^?1 (N3), 157.5 kg ha^?1 (N4), and 210 kg ha^?1 (N5)]. It was observed that N3 produced optimum total dry matter (5407 kg ha^?1), 1000 grain weight (39.7 g), grain yield (2.64 t ha^?1), and protein content (13.97%). Our results showed that N fertilization significantly increased photosynthetic parameters and N metabolite enzymes at all growth stages. Nitrogen harvest index, partial productivity factor, agronomic recovery efficiency, and nitrogen agronomic efficiency were decreased with increased N. Higher N rates (N3–N5) maintained higher photosynthetic capacity and dry matter accumulation and lower intercellular CO₂ content. The N supply influenced NUE by improving photosynthetic properties. The N3 produced highest chlorophyll content, photosynthetic rate, stomatal conductance and transpiration rate, grain yield, grain protein, dry matter, grains weight, and N metabolite enzyme activities compared to the other rates (N1, N2, N4, and N5). Therefore, increasing N rates beyond the optimum quantity only promotes vegetative development and results in lower yields.

     

Identification of TLR2/4-mediated phagocytosis and immune response activation pathways by vacuoles isolated from Saccharomyces cerevisiae

The vacuoles of the yeast Saccharomyces cerevisiae are closely related to mammalian lysosomes and play a role in macromolecular degradation due to the hydrolytic enzymes present inside. The vacuoles also regulate osmotic pressure and control cellular homeostasis. In previous results, vacuoles were shown to activate the immune response of macrophages by promoting the production of immune-mediated transporters nitric oxide (NO), reactive oxygen species (ROS), and pro-inflammatory cytokines. In this study, the effects of vacuoles on the phagocytosis activity of RAW264.7 cells and their potential as immune enhancers were evaluated, and receptors capable of recognizing vacuoles were examined. An investigation using the phagocytes assay showed that phagocytosis activity increased by the vacuole. Besides, after treatment with TLR2/4 inhibitor, the expression of pro-inflammatory cytokines by vacuoles was significantly reduced and the inducible nitric oxide synthase (iNOS) protein was also significantly reduced. However, treatment with a TLR2 inhibitor did not reduce the production of interleukin-6 (IL)-6, a pro-inflammatory cytokine. As a result of confirming the activation of TLR2/4 using Western blot and immunofluorescence (IF), the TLR2/4 protein expression and fluorescence intensity increased depending on the concentration of vacuoles. Yeast vacuoles significantly upregulate protein expression of p-p65/p-p38 MAPKs. In summary, the vacuoles isolated from S. cerevisiae in macrophages have increased phagocytic ability at a concentration of 20 (µg/ml) and can function as immune-enhancing agent suggesting that TLR2/4 mediated the p38 MAPK/nuclear factor kappa B signaling pathway.