Changes in selected stress and immune‐related genes in Atlantic cod,Gadus morhua,following overcrowding

Using Atlantic cod, Gadus morhua, as a model for the stress response in gadoid fish, the changes in the expression of some stress and immune genes as well as the profiles of plasma cortisol were examined. Adult fish were kept at a density of ca. 100 kg m^?3 by lowering the water level in the rearing tank for 1 h and this short‐term crowding stress was repeated thrice over a 12‐h interval period. Blood samples were collected before exposure and at 2, 24 and 72 h post crowding. Plasma cortisol level significantly increased at 2 h post crowding but returned to pre‐crowding levels 24 h after exposure. The relative expression of the stress response genes, glucose transporter‐3 and a putative heat shock protein 70 significantly increased at 2 and 24 h post crowding respectively. Significant up‐regulation of the pro‐inflammatory cytokines, interleukin (IL)‐1β and IL‐8, as well as anti‐bacterial genes, g‐type lysozyme and bactericidal permeability‐increasing protein/lipopolysaccharide‐binding protein (BPI/LBP) was also observed at 2 h and the levels were maintained until 72 h post exposure, except for BPI/LBP which had maximum up‐regulation at 24 h. The present observations have implications with respect to fish welfare and assessment of the health status of the farmed fish.     

外源褪黑激素对罕山白绒山羊绒毛生长相关基因表达差异的影响

试验旨在筛选罕山白绒山羊皮肤毛囊生长脱落相关的关键基因,并揭示外源褪黑激素（Melatonin,MT）对罕山白绒山羊绒毛生长相关基因表达差异的影响。选择10只体重平均为33.3 kg、年龄为20月龄的罕山白绒山为实验动物,分为2组,从2014年12月开始每隔1个月按照2 mg/（kg·BW）的剂量在埋植组绒山羊耳后皮下埋植MT,于2015年9月采集2组罕山白绒山羊皮肤组织进行RNA-Seq测序,对差异表达基因进行筛选、功能注释和富集分析。共筛选获得了475个在埋植组与对照组中差异表达的基因,其中,上调表达基因392个,下调表达基因83个。KEGG富集性分析表明,差异表达基因主要参与白细胞经内皮迁移、趋化因子信号通路、fcγ-R介导的吞噬作用、吞噬体、造血细胞谱系、补体级联和混凝级联、细胞因子—细胞因子受体相互作用、NF-kappa B信号通路、PI3K-Akt信号通路等。PI3K-Akt信号通路可能在褪黑激素调控皮肤毛囊发育中发挥重要作用。研究结果为进一步研究MT影响皮肤毛囊生长的分子机理提供依据。     

二丁基羟基甲苯对雨生红球藻虾青素和油脂积累的影响

以雨生红球藻(Haematococcus pluvialis)为对象,研究高光照、缺氮条件下,外源添加不同浓度的二丁基羟基甲苯(Butylated hydroxytoluene, BHT)对雨生红球藻生长、虾青素积累、油脂合成、脂肪酸组成、碳水化合物、蛋白含量以及虾青素和脂肪酸合成相关酶基因的影响。结果显示,添加不同浓度的BHT后,2 mg/L BHT添加组虾青素积累量为最高,显著高于其他实验组和对照组(P<0.05),达到31.66 mg/g,是对照组的1.87倍。油脂含量达45.56%,高于同期对照组(39.06%),脂肪酸组成变化不显著。在此条件下,虾青素合成关键酶基因dxs和bkt表达水平分别是对照组的5.19倍和2.04倍;脂肪酸合成关键酶基因kas和acp表达水平较对照组显著提高(P<0.05),分别是对照组的4.56倍和3.02倍。与对照组相比,2 mg/L BHT添加组的碳水化合物和蛋白含量均呈下降趋势。研究表明,在胁迫条件下,外源添加适量浓度的BHT能有效促进雨生红球藻中虾青素的积累,同时提高了藻细胞内的油脂含量。     

Sodium alginate supplementation modulates gut microbiota,health parameters,growth performance and growth‐related gene expression in Malaysian Mahseer Tor tambroides

This study investigated the effects of sodium alginate supplementation on gut microbiota composition, health parameters, growth performances and growth‐related gene expression of Malaysian mahseer. Five test diets were formulated by supplementing 0%, 0.1%, 0.2%, 0.4% and 0.8% sodium alginate. Triplicate groups of juvenile Tor tambroides (2.19 ± 0.05 g) were stocked in 15 aquaria (20 individuals per aquarium) and fed at 3.0% body weight per day for 60 days. PCoA and UPGMA analysis showed that gut bacterial community were more convergence in higher sodium alginate‐supplemented diets. The percentage of Porphyromonadaceae, Bacteroides, Plesiomonas and Shewanella were substantially higher and Aeromonas, Entomoplasmatales and Prevotellaceae were drastically lower in higher sodium alginate (0.2%–0.8%) diets. Sodium alginate supplementation (≥0.2%) significantly improved the haematocrit value and respiratory burst activity of T. tambroides. Growth performances and feed utilization were significantly higher in 0.2%–0.4% sodium alginate‐supplemented diets. The increased growth rate of T. tambroides was governed by both hyperplastic and hypertrophic muscle growth. Real‐time PCR data demonstrated that most of the growth‐related genes were significantly upregulated in 0.2%–0.4% sodium alginate‐supplemented diet. Finally, it can be concluded that sodium alginate should be supplemented at 2 g/kg in practical fish feed formulation.     

Effects of five compound attractants in high plant‐based diets on feed intake and growth performance of juvenile turbot (Scophthalmus maximus L.)

An experiment was conducted to evaluate the effects of five attractants on feed intake, growth performance and appetite‐regulating genes expression for juvenile turbot (Scophthalmus maximus L.). Fish were fed with seven diets: fish‐meal‐based diet (FM), fermented soybean meal replacing 45% fish meal (FSBM) and other diets with 1% compound attractant A (CA‐A) consisting of amino acids, compound attractant B (CA‐B) consisting of quaternary ammonium bases, compound attractant C (CA‐C) consisting of nucleotides, compound attractant G (CA‐G) consisting of the combination of CA‐A, CA‐B and CA‐C and blue mussel protein hydrolysate (BMH) on the basis of FSBM diet respectively. Daily feed intake in CA‐A group was significantly higher, but that in CA‐B group was significantly lower than FSBM group. Similarly, specific growth rate was significantly enhanced in CA‐A and CA‐G groups and were significantly decreased in CA‐B group. Protein efficiency ratio and protein retention in CA‐B group were significantly lower than FSBM group. Furthermore, BMH supplement increased the mRNA expression of Ghrelin in the intestine of fish. These results showed that CA‐A and CA‐G supplement could enhance growth in plant‐based diets, but CA‐B supplement had the opposite effect, which might be associated with lower feed intake and lower feed utilization.     

三角帆蚌外套膜表达的免疫相关基因筛选

利用三角帆蚌(Hyriopsis cumingii)外套膜cDNA文库EST序列,通过BLAST分析注释基因功能,发现35个与免疫防御功能相关的基因。根据其功能,这些免疫相关基因可划分为7类,即细胞免疫过程(2个)、蛋白酶和蛋白酶调节子(9个)、压力蛋白(7个)、抗菌肽(5个)、溶酶体酶(2个)、粘着蛋白(3个)及细胞凋亡和细胞周期调控相关基因(7个)。免疫相关基因在三角帆蚌外套膜中的广泛表达表明外套膜是重要的免疫组织。该研究为三角帆蚌外套膜分子免疫机理研究和抗病育种工作提供了基础性资料。     

Cloning and expression analysis of innate immune genes from red sea bream to assess different susceptibility to megalocytivirus infection

As suggested by the Office International des Epizooties (OIE), fishes belonging to the genus Oplegnathus are more sensitive to megalocytivirus infection than other fish species including red sea bream (Pagrus major). To assess the roles of the innate immune response to these different susceptibilities, we cloned the genes encoding inflammatory factors including IL‐8 and COX‐2, and the antiviral factor like Mx from red sea bream for the first time and performed phylogenetic and structural analysis. Analysed expression levels of IL‐1β, IL‐8 and COX‐2 and the antiviral factor like Mx genes performed with in vivo challenge experiment showed no difference in inflammatory gene expression or respiratory burst activity between red sea bream and rock bream (Oplegnathus fasciatus). However, the Mx gene expression levels in red sea bream were markedly higher than those in rock bream, suggesting the importance of type I interferon (IFN)‐induced proteins, particularly Mx, during megalocytivirus infection, rather than inflammation‐related genes. The in vitro challenge experiments using embryonic primary cultures derived from both fish species showed no difference in cytopathic effects (CPE), viral replication profiles, and inflammatory and Mx gene expression pattern between the two fish species.     

Soybean plant height QTL mapping and meta‐analysis for mining candidate genes

Plant height is closely related to seed yield of soybean. The goal of this study was to identify important loci affecting soybean plant height using meta‐analysis based on a reference physical map. Plant height related to QTLs was mapped across eight years with a RIL population by WinQTLCart v2.5. 182 QTLs related to plant height of soybean from database and our research were collected, and each QTL was projected onto the soybean physical map by software BioMercator v2.1. The confidence interval of meta‐QTL ranged from 0.09 to 5.07 Mb, and the mean phenotypic variance ranged from 4.9% to 73.0%. Furthermore, 4,259 candidate genes were located in these consensus QTLs, and 40 of them were involved in the plant growth and stem elongation and annotated as plant hormone signal transduction (pathway ID ko04075) in KEGG pathway. These results would lay a foundation for fine mapping of QTLs/genes related to plant height and marker‐assisted selection for breeding in soybean.     

层出镰刀菌处理对食用百合保护酶相关基因表达的影响

以4种食用百合为试材,利用实时荧光定量PCR(RT-qPCR)方法,研究了层出镰刀菌处理对食用百合保护酶相关基因(POD、CAT)相对表达量的影响,以期为研究食用百合抵抗逆境的分子机制提供参考依据。结果表明:POD在“龙牙×兰州”(R6-2、6)杂交百合、“铁炮×兰州”杂交百合、“兰州”百合被处理后的第1天相对表达量达到峰值且显著高于处理后的其它时间段,分别约为对照组的18.57、63.81、51.41、37.42倍,然而“铁炮”百合则是被处理后的第3天相对表达量达到峰值且显著高于处理后的其它时间段,约为对照组的113.19倍;CAT在“龙牙×兰州”(6)杂交百合、“铁炮×兰州”杂交百合均是被处理后的第1天相对表达量达到峰值且其显著高于处理后的其它时间段,分别约为对照组的68.02、19.77倍,在“龙牙×兰州”(R6-2)杂交百合、“兰州”百合则是被处理后的第3天相对表达量达到峰值,且显著高于处理后的其它时间段,分别约为对照组的30.69、25.19倍,铁炮百合是被处理后的第5天基因相对表达量达到峰值且显著高于其它时间段,约为对照组的71.83倍。     

细胞凋亡与疾病

细胞凋亡(apoptos is)是普遍存在于人体组织细胞内的细胞死亡的形式之一,是不同于坏死的正常生理性的程序性细胞死亡。细胞抗凋亡是生物机体为了适应环境变化,维持生理平衡的一种主动的自我保护行为。凋亡或抗凋亡均是在基因控制下,多样性、偶联性、多途径的信号转导过程。它也受到内外生存因子的影响和制约,肿瘤等多种疾病均能影响细胞凋亡过程。