植物淹水胁迫的生理学机制研究进展

淹水胁迫引起弱光环境,使气体扩散受限,叶片细胞膜脂过氧化加剧,体内保护酶系统受损,叶绿素降解,丙二醛含量积累,光合速率下降。为了适应淹水环境,植物通过生理生化机制的调节来保证淹水条件下的生命活动。如细胞通过调节渗透物质的含量来保持渗透势的平衡;细胞内各种抗氧化酶活性增加,以清除自由基,避免或者减轻细胞受到伤害;改变代谢途径和激素调节以保持能量储备和低的代谢速率。本文综述了淹水胁迫对细胞膜系统及功能、植物光合作用、植物呼吸、激素、生理代谢、基因调控的影响和淹水结束后植物的生理生态学变化,介绍了植物适应淹水胁迫的机制,并指出植物耐淹响应的分子机理,环境因素对淹没植物的影响,森林淹水胁迫的定位观测是今后需要研究的方面。     

淹水胁迫对不结球白菜根系生长与呼吸酶活性的影响

采用双套盆法,以不结球白菜‘新矮青’和‘新夏青2号’品种为材料,研究了不同时间(1、3、5、恢复7d后)和不同程度淹水处理(根淹、半淹)后不结球白菜根系生长及呼吸代谢的变化规律。结果显示:(1)与对照相比,淹水胁迫下,不结球白菜幼苗根系鲜重、根系长度、根系活力显著下降,且半淹处理的下降幅度大于根淹处理。(2)淹水胁迫下,乳酸脱氢酶(LDH)、乙醇脱氢酶(ADH)活性较对照显著升高,而苹果脱氢酶(MDH)、琥珀酸脱氢酶(SDH)活性显著降低,且半淹处理的降幅大于根淹,淹水5d后与淹水1d后有显著性差异。(3)淹水胁迫下,‘新矮青’乳酸发酵途径弱于‘新夏青2号’,乙醇发酵则相反,导致后者根系中乳酸积累多于前者,细胞质酸化较严重,降低了对淹水胁迫的耐性。研究表明,不结球白菜幼苗期受到淹水胁迫时,其有氧呼吸明显受阻,无氧呼吸代谢被促进,而且随着淹水时间的延长及淹水深度的加深,根系呼吸代谢受到的抑制程度越严重,最终导致根系生长受到抑制。     

基于转录组测序的扎龙湿地野大麦耐盐性分析

通过转录组测序技术对扎龙湿地野大麦幼苗在200 mmol/L NaCl胁迫2 d后的叶片进行分析。对测序结果进行De novo拼接后,将差异表达基因在GO、KEGG数据库中进行比对注释。结果表明:NaCl胁迫2 d后,对照组与处理组分别获得Unigene序列61038个和46754个,检测到差异表达基因25465个,差异基因GO功能注释到3个大类的55个功能组。差异表达基因被注释到135个Pathway上,直观地显示出NaCl胁迫下野大麦幼苗体内发生调节及改变的代谢过程和信号通路,其中主要涉及光合作用、脯氨酸代谢、叶绿素代谢等途径。通过对野大麦幼苗叶片转录组分析,发掘相关耐盐基因,有助于培育野大麦及其近缘作物新品种,并对揭示植物耐盐性分子机制及相关代谢途径具有重要意义。     

淹涝胁迫对鹅掌楸属植物叶片部分生理指标的影响

研究了淹涝胁迫对不同种源鹅掌楸[Liriodendron chinense（Hemsl．）Sarg．]和杂种鹅掌楸（L．chinense xL．tulipifera）幼苗部分生理指标的影响。结果表明：在淹水处理24h后,杂种鹅掌楸幼苗叶片的净光合速率变化不明显,鹅掌楸幼苗叶片的净光合速率则有一定程度的下降;在淹水处理48h后,净光合速率均大幅度下降。在淹涝胁迫下及胁迫解除后,幼苗叶片叶绿素含量持续缓慢下降;而脯氨酸含量总体上呈现上升的趋势;与鹅掌楸相比,杂种鹅掌楸幼苗的超氧化物歧化酶（SOD）和过氧化物酶（POD）活性变化不明显,但SOD活性最高,POD活性最低。结果表明,杂种鹅掌楸具有较强的耐淹涝胁迫的能力。     

东乡野生稻苗期响应低温胁迫的转录组分析

为了解东乡野生稻（Oryza rufipogon）对低温胁迫的响应机制,对苗期的RNA-seq转录表达谱进行了研究。结果表明,与对照相比,共检测到10 200个差异表达基因（DEGs）,其中5 201个上调表达,4 999个下调表达,其中有426个DEGs位于已报道的水稻耐冷QTL区间,且37个为耐冷调控相关的家族基因。GO功能分类和KEGG代谢路径分析表明,核酸结合转录因子活性、氨基酸生物合成以及光合作用代谢等均参与响应低温胁迫过程。实时荧光定量分析表明,ABA响应蛋白基因、MYB转录因子和40S核糖体蛋白SA基因等12个可能与低温胁迫响应相关的DEGs表达模式与RNA-seq的一致。可见,植物激素传导途径和转录因子相关调控基因在东乡野生稻苗期响应低温胁迫过程中起重要作用。     

阿尔泰蝠蛾(鳞翅目:蝙蝠蛾科)幼虫低温胁迫转录组分析

[目的]本研究旨在通过转录组测序技术分析低温胁迫引起的阿尔泰蝠蛾Hepialus altaicola Wang幼虫基因转录差异及上调表达基因的主要功能类群和参与的主要代谢通路.[方法]采用Illumina HiSeq TM 2500测序平台对阿尔泰蝠蛾幼虫进行转录组测序、组装,利用Blast软件进行数据库比对和基因功能注释,用DEGSeq R软件包分析4℃低温处理与室内适温饲养试虫的差异表达基因,并对上调表达基因进行GO和KEGG代谢途径富集分析.[结果]经序列拼接后共获得100300个unigenes,总长度81600309 bp,平均长度813 bp,N50长度1719 bp.与7大数据库同源比对,共获得34691(34.59％)条unigenes.低温胁迫转录组分析得到11569个差异表达基因(DEGs),7158条基因上调,4411条基因下调.富集到47个GO类群,217个KEGG途径.其中代谢过程、催化、结合活性类群占有重要比例,核糖体、碳代谢、剪接体等途径显著富集.另外,热激蛋白、昆虫表皮蛋白、海藻糖酶、超氧化物歧化酶等非生物胁迫相关基因显著上调表达.[结论]阿尔泰蝠蛾幼虫低温胁迫转录组分析揭示,代谢过程、细胞过程、生物调节、对刺激的反应等生物学过程相关基因和部分非生物胁迫响应基因显著上调表达,提示蝠蛾幼虫可能从抗氧化防御、分子伴侣、体温调节和维持细胞的渗透平衡等多方面应对低温胁迫.     

铁蛋白DrfE对耐辐射异常球菌抗氧化酶活性的影响

为研究耐辐射异常球菌(Deinococcus radiodurans)中drf E编码的铁蛋白Drf E的功能,通过基因回补试验构建drf E基因的回补菌株(pdrf E∷Δ),对野生型WT、突变株Δdrf E及回补菌株pdrf E∷Δ进行不同浓度H_2O_2和Na Cl胁迫,分别测定野生型WT、突变株Δdrf E和回补菌株pdrf E∷Δ体内Fe2+含量及抗氧化酶类活性。结果显示,drf E基因缺失导致菌株对氧化和盐胁迫敏感,该基因的回补能够恢复菌株对氧化和盐胁迫的抗性。drf E基因缺失导致耐辐射异常球菌体内Fe2+浓度由232μmol/L增加至293μmol/L;80 mmol/L H_2O_2处理30 min后,突变株Δdrf E的CAT活性下降32.74%,SOD下降41.3%。以上结果表明Drf E蛋白可能参与耐辐射异常球菌铁代谢途径,并且在细胞抗氧化体系中发挥重要作用。     

水稻OsCatB敲除突变体的构建及耐逆性初步分析

水稻过氧化氢酶B (catalase B, CatB)主要在非光合组织、愈伤组织、根和种子中表达,参与调控活性氧动态水平的平衡和根系的生长。目前, CatB参与非生物胁迫响应的功能在很大程度上是未知的。本研究利用基因编辑技术CRISPR/Cas9获得OsCatB基因敲除的纯合突变体catb,并分析了catb在盐、高温和氧化等胁迫处理下的生理和生化表型。研究结果发现,在盐处理下,突变体catb的生理和生化表型与野生型植株无显著差异;在热胁迫下, catb的过氧化氢酶活性和存活率与野生型植株相比显著降低,而H2O2含量显著升高;在氧化胁迫下, catb的幼苗高度低于野生型。这些结果说明,水稻CatB参与盐胁迫响应的调控不明显,而主要参与了高温和氧化胁迫响应,并正调控水稻的高温和氧化胁迫耐受性。该研究为进一步探究Os CatB参与逆境响应的分子机制和培育抗逆水稻品种提供了一定的理论指导和遗传材料。     

阿尔泰蝠蛾(鳞翅目:蝙蝠蛾科)幼虫低温胁迫转录组分析

[目的]本研究旨在通过转录组测序技术分析低温胁迫引起的阿尔泰蝠蛾Hepialus altaicola Wang幼虫基因转录差异及上调表达基因的主要功能类群和参与的主要代谢通路.[方法]采用Illumina HiSeq TM 2500测序平台对阿尔泰蝠蛾幼虫进行转录组测序、组装,利用Blast软件进行数据库比对和基因功能注释,用DEGSeq R软件包分析4℃低温处理与室内适温饲养试虫的差异表达基因,并对上调表达基因进行GO和KEGG代谢途径富集分析.[结果]经序列拼接后共获得100300个unigenes,总长度81600309 bp,平均长度813 bp,N50长度1719 bp.与7大数据库同源比对,共获得34691(34.59％)条unigenes.低温胁迫转录组分析得到11569个差异表达基因(DEGs),7158条基因上调,4411条基因下调.富集到47个GO类群,217个KEGG途径.其中代谢过程、催化、结合活性类群占有重要比例,核糖体、碳代谢、剪接体等途径显著富集.另外,热激蛋白、昆虫表皮蛋白、海藻糖酶、超氧化物歧化酶等非生物胁迫相关基因显著上调表达.[结论]阿尔泰蝠蛾幼虫低温胁迫转录组分析揭示,代谢过程、细胞过程、生物调节、对刺激的反应等生物学过程相关基因和部分非生物胁迫响应基因显著上调表达,提示蝠蛾幼虫可能从抗氧化防御、分子伴侣、体温调节和维持细胞的渗透平衡等多方面应对低温胁迫.     

灰葡萄孢丝裂原活化蛋白激酶编码基因bmp1和bmp3的功能

【背景】植物病原真菌丝裂原活化蛋白激酶(Mitogen-activated protein kinase,MAPK)信号途径参与病菌有性生殖、细胞壁完整、菌丝侵染、致病力、胁迫响应等过程,灰葡萄孢MAPK信号途径参与病菌生长发育、致病力以及胁迫响应,但MAPK信号途径基因在灰葡萄孢中的功能尚未完全阐明,该信号途径对灰葡萄孢的生长发育和致病力的调控机制尚不明确。【目的】明确灰葡萄孢MAPK编码基因bmp1、bmp3在病菌生长发育、致病力以及氧化胁迫响应过程的功能,为进一步阐明MAPK信号途径调控灰葡萄孢生长发育和致病力的分子机制奠定基础。【方法】利用RNAi技术构建灰葡萄孢MAPK编码基因bmp1和bmp3的RNAi突变体,并以野生型BC22菌株为对照,对bmp1和bmp3基因的RNAi突变体的表型、致病力以及对氧化胁迫的敏感性进行分析。【结果】灰葡萄孢bmp1和bmp3基因的RNAi突变体其菌落形态、菌丝形态均与野生型BC22菌株没有明显差别;bmp1基因的RNAi突变体生长速率明显减慢,分生孢子产量明显降低;bmp3基因的RNAi突变体的生长速率与野生型BC22菌株没有明显差别,不能产生分生孢子。bmp1和bmp3基因的RNAi突变体在番茄果实的表面均不能产生明显的致病症状,而且不能穿透玻璃纸。bmp1基因的RNAi突变体在含有H_2O_2的培养基上受抑制的程度显著低于野生型,而在含甲萘醌的培养基上受抑制的程度显著高于野生型;bmp3基因的RNAi突变体在含有H_2O_2和甲萘醌的培养基受抑制的程度均显著高于野生型。【结论】灰葡萄孢bmp1基因正调控病菌生长、分生孢子形成、致病力和穿透能力,参与调控病菌对氧化胁迫的响应;灰葡萄孢bmp3基因正调控病菌分生孢子形成、致病力、穿透能力以及对氧化胁迫的响应。     

Cytochrome P450 2E1 potentiates ethanol induction of hypoxia and HIF-1α in vivo

Ethanol induces hypoxia and elevates HIF-1α in the liver. CYP2E1 plays a role in the mechanisms by which ethanol generates oxidative stress, fatty liver, and liver injury. This study evaluated whether CYP2E1 contributes to ethanol-induced hypoxia and activation of HIF-1α in vivo and whether HIF-1α protects against or promotes CYP2E1-dependent toxicity in vitro. Wild-type (WT), CYP2E1-knock-in (KI), and CYP2E1 knockout (KO) mice were fed ethanol chronically; pair-fed controls received isocaloric dextrose. Ethanol produced liver injury in the KI mice to a much greater extent than in the WT and KO mice. Protein levels of HIF-1α and downstream targets of HIF-1α activation were elevated in the ethanol-fed KI mice compared to the WT and KO mice. Levels of HIF prolyl hydroxylase 2, which promotes HIF-1α degradation, were decreased in the ethanol-fed KI mice in association with the increases in HIF-1α. Hypoxia occurred in the ethanol-fed CYP2E1 KI mice as shown by an increased area of staining using the hypoxia-specific marker pimonidazole. Hypoxia was lower in the ethanol-fed WT mice and lowest in the ethanol-fed KO mice and all the dextrose-fed mice. In situ double staining showed that pimonidazole and CYP2E1 were colocalized to the same area of injury in the hepatic centrilobule. Increased protein levels of HIF-1α were also found after acute ethanol treatment of KI mice. Treatment of HepG2 E47 cells, which express CYP2E1, with ethanol plus arachidonic acid (AA) or ethanol plus buthionine sulfoximine (BSO), which depletes glutathione, caused loss of cell viability to a greater extent than in HepG2 C34 cells, which do not express CYP2E1. These treatments elevated protein levels of HIF-1α to a greater extent in E47 cells than in C34 cells. 2-Methoxyestradiol, an inhibitor of HIF-1α, blunted the toxic effects of ethanol plus AA and ethanol plus BSO in the E47 cells in association with inhibition of HIF-1α. The HIF-1α inhibitor also blocked the elevated oxidative stress produced by ethanol/AA or ethanol/BSO in the E47 cells. These results suggest that CYP2E1 plays a role in ethanol-induced hypoxia, oxidative stress, and activation of HIF-1α and that HIF-1α contributes to CYP2E1-dependent ethanol-induced toxicity. Blocking HIF-1α activation and actions may have therapeutic implications for protection against ethanol/CYP2E1-induced oxidative stress, steatosis, and liver injury.     

Upregulation of transcription factor NRF2-mediated oxidative stress response pathway in rat brain under short-term chronic hypobaric hypoxia

Exposure to high altitude (and thus hypobaric hypoxia) induces electrophysiological, metabolic, and morphological modifications in the brain leading to several neurological clinical syndromes. Despite the known fact that hypoxia episodes in brain are a common factor for many neuropathologies, limited information is available on the underlying cellular and molecular mechanisms. In this study, we investigated the temporal effect of short-term (0–12 h) chronic hypobaric hypoxia on global gene expression of rat brain followed by detailed canonical pathway analysis and regulatory network identification. Our analysis revealed significant alteration of 33, 17, 53, 81, and 296 genes (p < 0.05, <1.5-fold) after 0.5, 1, 3, 6, and 12 h of hypoxia, respectively. Biological processes like regulation, metabolic, and transport pathways are temporally activated along with anti- and proinflammatory signaling networks like PI3K/AKT, NF-κB, ERK/MAPK, IL-6 and IL-8 signaling. Irrespective of exposure durations, nuclear factor (erythroid-derived 2)-like 2 (NRF2)-mediated oxidative stress response pathway and genes were detected at all time points suggesting activation of NRF2-ARE antioxidant defense system. The results were further validated by assessing the expression levels of selected genes in temporal as well as brain regions with quantitative RT-PCR and western blot. In conclusion, our whole brain approach with temporal monitoring of gene expression patterns during hypobaric hypoxia has resulted in (1) deciphering sequence of pathways and signaling networks activated during onset of hypoxia, and (2) elucidation of NRF2-orchestrated antioxidant response as a major intrinsic defense mechanism. The results of this study will aid in better understanding and management of hypoxia-induced brain pathologies.