Resveratrol protects cortical neurons in infant rats with bacterial meningitis through miR-223-3p/NLRP3 pathway

AIM To investigate the protective effect of resveratrol (Res) on cortical neurons in rat bacterial meningitis (BM) model. METHODS Group B hemolytic Streptococcus was injected via the posterior cistern to establish a BM model. Resveratrol was administered intranasally and microRNA-223-3p (miR-223-3p) antagomir was administered by intracerebroventricular injection. HE staining was used to observe the pathological changes of the brain tissue. Loeffler scoring method was used to evaluate the neurobehavioral functions. TUNEL staining was used to detect neuronal apoptosis. The expression of interleukin-1β (IL-1β), IL-18, glial fibrillary acidic protein (GFAP) and ionized calcium-binding adaptor molecule 1 (Iba1) was detected by immunofluorescence staining. The protein levels of nucleotide-binding oligomerization domain-like receptor protein 3 （NLRP3）, cleaved caspase-1, IL-1β and IL-18 were determined by Western blot. The expression level of miR-223-3p was detected by RT-qPCR. Online software TargetScan was used to search for the complementary nucleotide sequences between miR-223-3p and NLRP3 mRNA. RESULTS Compared with sham group, the thickness of meninges in BM model was increased, the neurological score was decreased (P<0.05), and the number of TUNEL positive neurons was increased significantly (P<0.05). Astrocytes and microglia were activated, the fluorescence intensity of IL-1β and IL-18 was increased (P<0.05), and the expression levels of NLRP3, cleaved caspase-1, IL-1β, IL-18 and miR-223-3p were increased (P<0.05). Compared with BM group, after treatment with resveratrol, the neurological score was increased (P<0.05), the number of TUNEL positive neurons was decreased significantly (P<0.05), and the inflammatory response of astrocytes and microglia was suppressed. The fluorescence intensity of IL-1β and IL-18 was decreased (P<0.05), the protein levels of NLRP3, cleaved caspase-1, IL-1β and IL-18 were decreased (P<0.05), and the expression level of miR-223-3p was increased (P<0.05). A nucleotide sequence in the 3'-UTR of NLRP3 mRNA might be targeted by miR-223-3p. In the brain of rat BM model, compared with antagomir control group, the expression of NLRP3 was increased in miR-223-3p antagomir group with resveratrol treatment (P<0.05). CONCLUSION Resveratrol may reduce the inflammatory death of cortical neurons in BM model of infant rats through miR-223-3p/NLRP3 pathway, thus playing a protective role for the neurons.     

基于S基因序列分析新型猪流行性腹泻病毒的遗传演化

最近，笔者实验室在青藏高原地区发现两种新亚型藏猪源猪流行性腹泻病毒（PEDV），为进一步调查新型PEDV是否在四川腹泻猪群中存在或流行，对实验室2018-2019年保存的116份猪腹泻粪便或肠组织样本进行PEDV的检测及其纤突蛋白基因（spike）分子特征研究。结果表明：腹泻样本的PEDV检出率为42.2%（49/116，95% CI=33.1%~51.8%），并获得了13条完整的S基因序列，全长为4 149~4 170 bp，序列相似性为94.2%~99.9%，其中SWUN-H3-CH-SCYA-2019的S基因与藏猪源新G1亚群PEDV的序列相似性高达97.0%~98.6%。遗传演化研究结果表明13株PEDV S基因划分为G1和G2大群，其中SWUN-H3-CH-SCYA-2019位于藏猪源新G1亚群；SWUN-19-CH-SCZY-2018、SWUN-4-CH-SCXC-2018、SWUN-1-CH-SCNJ-2019和SWUN-3CH-CH-SCZG-2019位于G2亚群中一个独立的分支，且与藏猪源新G2亚群毒株有着较近的亲缘关系。为了进一步研究13株PEDV的演化过程，以贝叶斯进化分析软件包（BEAST）进行分歧时间估算，结果表明SWUN-H3-CH-SCYA-2019的分歧时间约为2012.3年，早于藏猪源新G1亚群其余毒株的最早分歧时间（2015.7年）；SWUN-4-CH-SCXC-2018、SWUN-19-CH-SCZY-2018和SWUN-3CH-CH-SCZG-2019的分歧时间约为2014.2年，早于G2亚群的藏猪源毒株2014.7年，所有藏猪源PEDV的分歧时间均晚于四川毒株。本研究在四川地区首次发现了藏猪源PEDV，并且从毒株的分歧时间推断青藏高原的藏猪源PEDV来源于四川，为新型PEDV分子遗传进化的监测提供了依据。     

花生种子特异启动子AHSSP1的克隆及功能分析

为丰富花生种子特异启动子资源,本研究利用PCR技术在花生基因组中克隆了种子贮藏蛋白基因PSC32的启动子AHSSP1,利用半定量RT-PCR检测了PSC32基因表达模式,借助NewPLACE在线分析了AHSSP1序列中存在的顺式作用元件,并构建了AHSSP1驱动GUS报告基因的表达载体,经农杆菌转化获得转基因拟南芥,经GUS组织化学染色鉴定了该启动子的功能。结果表明,PSC32基因957 bp长的启动子AHSSP1序列具备种子特异表达启动子特有的3个RY REPEAT元件。半定量RT-PCR分析发现,PSC32基因在花生成熟种子中表达,而在饱果成熟期根、茎、叶片、花、入土前的果针、成熟种子的果壳中均不表达。GUS组织化学染色发现,转基因拟南芥成熟种子以及萌发种子的子叶、下胚轴和胚根均能够被染上蓝色;长出真叶后,子叶和下胚轴仍能被染色,而根和真叶不能被染上蓝色;成年期转基因拟南芥的叶片也不能被染上蓝色。而野生型拟南芥整个生长时期均不能被染上蓝色。以上现象说明AHSSP1是一个种子特异启动子。本研究丰富了花生种子特异启动子的资源,对花生籽仁品质改良或以花生籽仁作为“生物反应器”的研究具有重要的应用价值。     

我国楠属种质资源分布现状及主要种特征差异

对福建、江西、浙江、湖南、湖北、贵州、重庆、四川、海南、云南及广西11省(自治区、直辖市)70个县(区、市)的楠属(Phoebe)树种种质资源进行详细调查,记录楠属种质资源分布、群落现状、木材利用及人工造林情况,对种质资源进行收集保存,并详细介绍主要树种的特征差异。调查发现,中国楠属树种资源主要分布在长江流域及以南地区,其中紫楠(P.sheareri)、闽楠(P.bournei)及白楠(P.neurantha)分布范围最广;综合比较生长速度和适应性,广西适宜推广种植的楠属树种有闽楠、桢楠(P.zhennan)及崖楠(P.yaiensis)。     

河北省优质棉育种70年回顾与展望

河北省优质陆地棉品种选育工作起步较早，新中国成立以来先后选育出石短5号、冀棉22号、冀棉26、新陆中8号、冀228、农大棉13号等高品质棉花品种和大量优质棉品种。根据不同时期的优质棉评价标准，对1949―2019年河北省选育的优质棉品种的纤维品质性状进行了统计和分析，结合纺织工业对优质原棉的需求，指出河北省优质棉育种存在的问题，并对未来的发展方向进行了展望。     

灰霉菌侵染对番茄幼苗活性氧积累及抗氧化酶活性的影响

为进一步分析活性氧和抗氧化防御系统在番茄抗病信号调控机制中的作用,为提高产量和品质提供科学依据,以4叶1心的番茄幼苗为试验材料,比较番茄幼苗受灰霉菌侵染后叶片O_2~-·以及抗氧化酶、内源褪黑素等的变化。结果表明:番茄幼苗接种灰霉菌后,随着时间的延长,幼苗叶片上病斑斑点逐渐增多,灰霉菌含量显著增多,叶色逐渐变浅,植株表现明显的发病症状;灰霉病菌侵染后番茄体内O_2~-·活性爆发,超氧化物歧化酶(SOD)和过氧化物酶(POD)活性下降,过氧化氢酶(CAT)和谷胱甘肽还原酶(GR)活性上升,褪黑素含量增加;上述变化都是番茄幼苗为了抵御灰霉菌的生物胁迫而发生的应激反应。综上,植物在遭受病原菌等生物胁迫时不能简单地推断SOD、CAT和POD等抗氧化酶活性一定升高,植物抵御病原菌侵染更多地依赖于抗性信号路径中信号物质的激活和积累。     

王氏唇柱苣苔的离体培养及遗传稳定性

以王氏唇柱苣苔(Chirita wangiana)叶片为外植体进行组培离体快繁研究,并利用流式细胞术对组培苗进行遗传稳定性分析。结果表明:最佳初代诱导培养基为MS添加0.5 mg/L 6-BA+0.1 mg/L NAA;最适继代培养基为MS添加0.5 mg/L 6-BA+0.05 mg/L NAA;最适生根培养基为1/2 MS培养基添加0.5 mg/L IBA和15 g/L蔗糖,所有生根培养基获得的组培苗移栽驯化成活率达到92%以上。组织学切片检测表明,王氏唇柱苣苔叶片为外植体所形成芽体均为器官发生方式形成。流式细胞术检测表明,组培苗倍性没有变化,基因组大小与母本相比仅发生了1.86%的减少;染色体数目为2n=36,跟母本一致,植株形态特征上也无变异。研究结果可为王氏唇柱苣苔的园艺应用快速提供大量遗传稳定的种苗。     

Root architecture of sorghum genotypes differing in root angles under different water regimes

Plant root architecture offers the potential for increasing soil water accessibility, particularly under water-limited conditions. The objectives of this study were to evaluate the root architecture in two genotypes of sorghum (Sorghum bicolor (L.) Moench) differing in root angles and to assess the influence of different deficit irrigation regimes on root architecture. The response of two sorghum genotypes, ‘Early Hegari-Sart’ (EH; steep root angle) and ‘Bk7’ (shallow root angle) to four irrigation treatments was investigated in two replicated outdoor studies using large pots. The results indicated that EH possessed steeper brace and crown root angles, fewer brace roots, greater root biomass, and root length density than Bk7 at deeper soil depths (i.e., 15–30 and 30–45 cm) compared with a shallower depth (i.e., 0–15 cm). Across the soil profile, EH had greater root length density and length of roots of small diameter (<1 mm) than Bk7. Accordingly, EH showed more rapid soil-water capture than Bk7. Different levels of irrigation input greatly affected root architecture. Severe deficit irrigation (25% of full crop transpiration throughout the season) increased the angle and number of crown roots, root biomass, and root length density compared with 75 and 100% of full crop transpiration treatments. Consequently, root system architecture can be effectively manipulated through both genotypic selection and irrigation management to ensure optimal performance under different levels of soil available water.     

农村老龄化下风险偏好、人际关系与农户化学品施用行为分析

基于441份农户的微观调查样本,从各类化肥使用量、农药使用量及其购买成本方面考察了农村劳动力老龄化对化学品投入的影响,并从老年劳动力的体力状况、控制感、技能需求、风险态度和人际关系等途径探究其影响机制,进一步考察农村老年劳动力个体特征对化学品投入产生的异质性影响。结果表明,农村劳动力老龄化会显著减少对复合肥的使用量以及化肥的购买成本。当控制了体力和技能需求等变量后,老龄化对化学品投入依旧有显著的负向影响,表明以往研究推测的体力和技术采纳是老龄化影响务农的途径,不能全面揭示其影响机制。进一步研究发现,农村劳动力老龄化会影响农户的人际关系以及风险偏好对务农投入的边际效应。结论表明,中国建设生态宜居的美丽乡村政策对国家粮食安全战略具有促进作用。