盐胁迫下宁夏枸杞Na+吸收及Na+/H+转运蛋白与H+-ATPase基因表达的研究

为从分子水平揭示宁夏枸杞钠的吸收积累机理,本试验采用原子吸收分光光度法和实时荧光定量PCR(RT-qPCR)方法,对盐胁迫下宁夏枸杞根中Na⁺、K⁺含量以及质膜和液泡膜Na⁺/H⁺转运蛋白与H⁺-ATPase基因表达水平进行测定分析。结果表明,相同胁迫时间下,随着NaCl处理浓度的增加,枸杞根系中Na⁺浓度总体呈缓慢增加趋势,K⁺含量呈先增加后减少趋势,Na⁺/K⁺比值呈先减少后增加趋势;编码质膜和液泡膜的Na⁺/H⁺转运蛋白基因LbSOS1、LbNHX1以及液泡膜H⁺-ATPase基因LbVHA-C1表达量均呈升高趋势,质膜H⁺-ATPase基因LbHA1表达量呈先升高后降低趋势。相同NaCl处理浓度下,随着胁迫时间的延长,Na⁺含量总体呈增加趋势,K⁺含量呈先增加后减少趋势,Na⁺/K⁺比值呈增加趋势。LbSOS1、LbNHX1表达量总体呈先升高后降低趋势,LbVHA-C1、LbHA1表达量总体呈降低的趋势。相关性分析显示,不同胁迫时间下,枸杞根中LbSOS1、LbNHX1、LbVHA-C1和LbHA1表达量与Na⁺含量存在一定的正相关或负相关。上述结果表明,在低浓度NaCl胁迫时,维持枸杞体内较高的K⁺/Na⁺比值是宁夏枸杞耐盐的主要方式之一,同时也说明在胁迫初期,质膜和液泡膜的Na⁺/H⁺转运蛋白与H⁺-ATPase参与了枸杞细胞中Na⁺及时排出胞外和区隔于液泡,从而保持了根细胞内Na⁺的稳定性。此外,随着胁迫时间的延长和NaCl处理浓度的增加,LbSOS1、LbNHX1、LbVHA-C1和LbHA1的表达水平均降低,而 Na⁺积累量大幅增加,致使枸杞抗盐性降低。本研究揭示了宁夏枸杞的耐盐机理,为利用宁夏枸杞改良宁夏大面积盐碱地提供了理论依据。     

碱胁迫下榆叶梅差异基因表达分析

为了解榆叶梅(Prumus triloba Lindl.)在短期碱胁迫下的分子响应机制,本研究采用Illumina HiSeq^TM2500高通量测序技术对其在0、1、3、6、12 h碱胁迫下的叶片进行了de novo转录组测序,以此鉴定与碱胁迫相关的差异表达基因(DEGs)。结果表明,从5 960万条raw reads中获得了约5 300万条高质量clean reads,经从头组装后获得了124 786条榆叶梅unigenes。基于8 948个 unigenes差异表达(DEGs), GO富集分析发现,其中有28个基因可能在早期碱胁迫响应机制中起着重要作用; KEGG富集分析发现,不同碱处理时间下的KEGG途径(pathways)具有显著差异。对7个碱性胁迫反应相关基因进行RT-qPCR验证,所得基因的表达模式与RNA-Seq数据结果一致,证实了RNA-Seq结果的可靠性。本研究首次对碱胁迫下的榆叶梅叶片进行了转录组分析,获得的序列数据极大地丰富了榆叶梅可利用的基因资源,深入了解了榆叶梅碱胁迫的分子响应机制,同时为研究以榆叶梅为砧木的中国李(Prunus salicina)在碱胁迫下的适应机制奠定了分子研究基础。     

谷子MDH基因非生物逆境响应特性研究

为揭示谷子MDH基因对逆境胁迫的响应,本研究通过序列比对得到2个谷子苹果酸脱氢酶(malate dehydrogenase, MDH)基因SiMDH1、SiMDH2,并对SiMDH1和SiMDH2的编码蛋白特征、基因结构、功能、进化等性状进行了系统的分析和预测,用荧光实时定量PCR(qPCR)检测了SiMDH1和SiMDH2在苗期不同逆境及关键生育期干旱胁迫和不同光照强度下的表达。蛋白特征参数和序列分析表明,SiMDH1和SiMDH2蛋白特征参数相似度很高,序列非常保守,都含有MDH基因的特征功能域苹果酸酶NAD结合域和苹果酸酶C端功能域。亚细胞定位预测显示,SiMDH1和SiMDH2主要定位于叶绿体、细胞质和线粒体。启动子区域顺式元件分析发现,在SiMDH1和SiMDH2启动子区域含有大量光应答、激素类和其他类应答元件。苗期逆境qPCR表达谱分析发现,SiMDH1和SiMDH2受脱落酸(ABA)、聚乙二醇(PEG)、高盐和低温不同程度诱导,揭示SiMDH1和SiMDH2参与了谷子苗期非生物逆境胁迫应答。进一步分析发现,SiMDH1和SiMDH2参与了谷子在拔节、抽穗、灌浆期的干旱应答,而光照强度会显著影响SiMDH1和SiMDH2基因在不同生育期的表达。本研究为进一步分析MDH逆境应答机制以及利用基因工程方法改善作物抗逆性提供了试验和理论支持。     

Surplus spermine does not protect salmon cells against lipopolysaccharide (LPS)‐induced stress,but excess spermine is eliminated through spermidine/spermine N1‐acetyltransferase (SSAT)

The present trials tested the efficiency of surplus spermine to reduce inflammation and oxidative stress following LPS‐induced stress using an in vitro model of head kidney and liver cells isolated from Atlantic salmon. Spermine did not protect cells from LPS‐induced inflammatory response at either 0.3, 0.6 or 0.9 mM. However, as the gene expression of spermidine/spermine N1‐acetyltransferase (SSAT) increased with increasing spermine concentration, we addressed possible oxidative effects of the increased SSAT using its activator DENSPM or inhibitor of polyamine oxidation of the acetylated polyamines using MDL72527 at a spermine concentration of 0.6 mM. There was no significant effect of DENSPM, but MDL72527 decreased gene expression of GPX‐3 (p = .04), while gene expression of catalase and MnSOD was unaffected by treatment (p = .30 and p = .48, respectively). In conclusion, spermine did not protect cells from LPS‐provoked inflammation. The higher the spermine concentration, the more SSAT producing acetylated spermine occurred. Inhibiting the acetylated polyamine oxidases by MDL72527 improved oxidation status as expected due to a lower endogenous production of H₂O₂ by polyamine and acetylated polyamine oxidases. Probably care should be taken using polyamines or arginine as functional ingredients to avoid any increased oxidation within cells.     

Effects of ferulic acid on growth performance,immunity and antioxidant status in genetically improved farmed tilapia (Oreochromis niloticus) fed oxidized fish oil

A study was conducted to characterize the effects of dietary oxidized fish oil on the growth performance, immunity and antioxidant status of genetically improved farmed tilapia (Oreochromis niloticus) and to determine the role of ferulic acid on the oxidative damage induced by the oxidized fish oil. The tilapia (13.73 ± 0.31 g) were fed four experimental diets containing untreated (peroxide value, POV: 2.2 meq/kg) and highly oxidized (POV: 120.6 meq/kg) fish oil either with or without ferulic acid (0 or 400 mg/kg) supplementation for 12 weeks. From the results, the oxidized fish oil treatments increased antioxidant enzyme activities and MDA values but decreased the weight gain and the immunological parameters in tilapia. Meanwhile, the serum biochemical indices were significantly affected by the oxidized fish oil. Besides, the addition of ferulic acid partially counteracted the free radical‐induced damage and improved the health status of tilapia. In conclusion, the oxidized fish oil may induce oxidative stress, destroy liver, dysregulate lipid metabolism as well as reduce non‐specific immunity, and eventually result in growth inhibition of tilapia. The ferulic acid supplementation partially offset the negative effects of the oxidized fish oil on tilapia.     

硝化应激在神经型犬瘟热疾病发展中的临床意义

为了研究硝化应激在神经型犬瘟热疾病发展中的作用,并探讨其对该病的诊断价值,选取犬瘟热临床病例113例,其中有神经症状组(NCD)共37例,无神经症状组(NNS)共76例,并设25例无犬瘟热健康犬作为对照组(CG)。就诊时分别采取血液和脑脊液,测定一氧化氮(NO)、神经元型一氧化氮合成酶(nNOS)和神经元特异性烯醇化酶(NSE)水平。结果显示:NCD组2种组织液中的NO、nNOS和NSE升高,显著高于NNS组(P<0.05)与CG组(P<0.01);ROC曲线分析发现,2种组织液中的NO、nNOS和NSE对神经型犬瘟热的诊断具有良好的灵敏度和特异性,血浆中以nNOS最佳,截断值为230.1 ng/mL,脑脊液中以NSE最佳,截断值为3.67 ng/mL;NSE对神经型犬瘟热具有一定预后价值,血浆中NSE<2.07的病例死亡率显著低于NSE≥2.07的病例。结果表明:神经型犬瘟热疾病发展中伴有硝化应激和神经损伤,血浆与脑脊液中的NO、nNOS和NSE具有对神经型犬瘟热的诊断及预测作用。     

Effects of acute environmental and handling exposure on physiology and fillet quality of market‐sized Channel Catfish

Catfish Ictalurus spp. are subjected to stressful conditions during harvest, which may be linked to fillet coloration and quality. Poor water quality in ponds, socks or hauling tanks, as well as handling stress, have been suggested to cause red fillets in catfish; however, chronic exposure has not resulted in red fillets. Short‐term occurrences of extreme poor water quality, particularly low dissolved oxygen, high carbon dioxide and high temperature, may occur in ponds or during harvest. Therefore, market‐sized Channel Catfish Ictalurus punctatus were acutely exposed (12 hr) to one of the three water quality treatments while confined during a simulated socking procedure and evaluated for stress responses by means of change in blood parameters and fillet quality. In fish subjected to the extreme treatment, hematocrit, plasma cortisol, glucose and lactate levels increased, with 22% mortality, indicating highly stressful conditions. In fish subjected to moderate and typical (control) treatments, cortisol increased but a lack of change or decrease in glucose and lactate indicated minimal anaerobic metabolism. Only one red fillet was produced by the extreme treatment and two by the typical treatment; therefore, the results suggest red fillets are not a product of poor water quality compounded by handling during harvest.     

接种根瘤菌后3个紫花苜蓿品种耐盐性综合评价

研究苜蓿中华根瘤菌共生3个耐盐性高的紫花苜蓿品种,以选育适宜在内蒙古盐渍土地区大量种植的紫花苜蓿品种。以内蒙古地区常见的3个紫花苜蓿品种(中苜1号、金皇后、阿尔冈金)为试验材料,利用不同浓度NaCl溶液(0、50、150 mmol/L)模拟盐害环境,对3个紫花苜蓿品种接种苜蓿中华根瘤菌,测定紫花苜蓿品种的地上部分鲜质量、根鲜质量、根瘤数、有效根瘤数、丙二醛(MDA)质量摩尔浓度、可溶性糖质量分数6项指标,依据隶属函数法进行耐盐性综合评价。当盐浓度达到50 mmol/L时,3个紫花苜蓿品种中金皇后与苜蓿中华根瘤菌共生耐盐效果最好,相比于不接种地上部分鲜质量增加6.14 g,MDA质量摩尔浓度低至0.91mmol/g,可溶性糖质量分数高达21.65 mg/g,在3个紫花苜蓿品种中膜脂过氧化程度最低,渗透调节能力最强。当盐浓度达到150 mmol/L时,接种苜蓿中华根瘤菌对3个紫花苜蓿品种影响微弱。耐盐性强弱依次为金皇后(接种RM)、中苜1号(接种RM)、阿尔冈金(接种RM)、中苜1号(未接种RM)、金皇后(未接种RM)、阿尔冈金(未接种RM)。不同盐浓度(0、50、150 mmol/L)对3个紫花苜蓿品种生长均有抑制作用,且抑制效果随盐浓度升高而增强。在轻度盐害(50 mmol/L)环境下,接种苜蓿中华根瘤菌有效缓解盐对紫花苜蓿的伤害,其中金皇后耐盐性最强。当重度盐害(150 mmol/L)发生时,接种苜蓿中华根瘤菌对紫花苜蓿作用效果不明显甚至不起作用。     

盐胁迫对羽衣甘蓝种子萌发的影响

为探明盐胁迫对羽衣甘蓝种子萌发特性的影响，明确种子萌发期对盐胁迫的耐受能力，本试验以羽衣甘蓝种子为试验材料，研究不同浓度的NaCL（0、40、80、120、160、200mmol/l）和NaHCO（3 0、20、30、40、50、60mmol/l）对种子萌发胁迫的作用。结果表明:1、2 种盐胁迫处理下，羽衣甘蓝种子的发芽率、发芽势、发芽指数和活力指数等指标均随着盐浓度的升高逐渐降低；2、高浓度的NaHCO³盐溶液对羽衣甘蓝种子的胚根、胚芽的生长抑制较NaCL 盐溶液显著；3、2 种盐