Genome-Wide Exploration of the Grape GLR Gene Family and Differential Responses of VvGLR3.1 and VvGLR3.2 to Low Temperature and Salt Stress

Grapes,one of the oldest tree species globally,are rich in vitamins.However,environmental conditions such as low temperature and soil salinization significantly affect grape yield and quality.The glutamate receptor(GLR)family,comprising highly conserved ligand-gated ion channels,regulates plant growth and development in response to stress.In this study,11 members of the VvGLR gene family in grapes were identified using whole-genome sequence analysis.Bioinformatic methods were employed to analyze the basic physical and chemical properties,phylogenetic trees,conserved domains,motifs,expression patterns,and evolutionary relationships.Phylogenetic and collinear analyses revealed that the VvGLRs were divided into three subgroups,showing the high conservation of the grape GLR family.These members exhibited 2 glutamate receptor binding regions(GABAb and GluR)and 3-4 transmembrane regions(M1,M2,M3,and M4).Real-time quantitative PCR analysis demonstrated the sensitivity of all VvGLRs to low temperature and salt stress.Subsequent localization studies in Nicotiana tabacum verified that VvGLR3.1 and VvGLR3.2 proteins were located on the cell membrane and cell nucleus.Additionally,yeast transformation experiments confirmed the functionality of VvGLR3.1 and VvGLR3.2 in response to low temperature and salt stress.Thesefindings highlight the significant role of the GLR family,a highly conserved group of ion channels,in enhancing grape stress resistance.This study offers new insights into the grape GLR gene family,providing fundamental knowledge for further functional analysis and breeding of stress-resistant grapevines.

Synergistic effects of carbon cycle metabolism and photosynthesis in Chinese cabbage under salt stress

Chinese cabbage(Brassica rapa ssp. pekinensis) has a long cultivation history and is one of the vegetable crops with the largest cultivation area in China. However, salt stress severely damages photosynthesis and hormone metabolism, nutritional balances, and results in ion toxicity in plants. To better understand the mechanisms of salt-induced growth inhibition in Chinese cabbage, RNA-seq and physiological index determination were conducted to explore the impacts of salt stress on carbon cycle metabolism and photosynthesis in Chinese cabbage. Here, we found that the number of thylakoids and grana lamellae and the content of starch granules and chlorophyll in the leaves of Chinese cabbage under salt stress showed a time-dependent response, first increasing and then decreasing. Chinese cabbage increased the transcript levels of genes related to the photosynthetic apparatus and carbon metabolism under salt stress, probably in an attempt to alleviate damage to the photosynthetic system and enhance CO_(2) fixation and energy metabolism. The transcription of genes related to starch and sucrose synthesis and degradation were also enhanced;this might have been an attempt to maintain intracellular osmotic pressure by increasing soluble sugar concentrations. Soluble sugars could also be used as potential reactive oxygen species(ROS) scavengers, in concert with peroxidase(POD)enzymes, to eliminate ROS that accumulate during metabolic processes. Our study characterizes the synergistic response network of carbon metabolism and photosynthesis under salt stress.

Melatonin Alleviates Abscisic Acid Deficiency Inhibition on Photosynthesis and Antioxidant Systems in Rice under Salt Stress

Melatonin and abscisic acid,as major plant hormones,play important roles in the physiological and biochemical activities of crops,but the interaction between the two under salt stress is not yet clear.This study investigated the endogenous levels of melatonin and abscisic acid in rice by using exogenous melatonin,abscisic acid,and their synthetic inhibitors,and examined their interactions under salt stress.The research results indicate that melatonin and abscisic acid can improve rice salt tolerance.Melatonin alleviated the salt sensitivity caused by abscisic acid deficiency,increased antioxidant enzyme activity and antioxidant content in rice treated with abscisic acid synth-esis inhibitors,and reduced total reactive oxygen species content and thiobarbituric acid reactive substance accu-mulation.Melatonin also increased the activity of key photosynthetic enzymes and the content of photosynthetic pigments,maintaining the parameters of photosynthetic gas exchange and chlorophyllfluorescence.In summary,melatonin alleviated the effects of abscisic acid deficiency on photosynthesis and antioxidant systems in rice and improved salt tolerance.This study is beneficial for expanding the understanding of melatonin regulation of crop salt tolerance.

Coseismic Coulomb stress changes induced by a 2020-2021 M_(W)>7.0 Alaska earthquake sequence in and around the Shumagin gap and its influence on the Alaska-Aleutian subduction interface

Three M_(W)>7.0 earthquakes in 2020-2021 occurred in the Shumagin seismic gap and its adjacent area of the Alaska-Aleutian subduction zone,including the Mw7.8 Simeonof thrust earthquake on July 22,2020,the M_(W)7.6 Sand Point strike-slip earthquake on October 19,2020,and the M_(W)8.2 Chignik thrust earthquake on July 29,2021.The spatial and temporal proximity of these three earthquakes prompts us to probe stress-triggering effects among them.Here we examine the coseismic Coulomb stress change imparted by the three earthquakes and their influence on the subduction interface.Our results show that:(1)The Simeonof earthquake has strong loading effects on the subsequent Sand Point and Chignik earthquakes,with the Coulomb stress changes of 3.95 bars and 2.89 bars,respectively.The Coulomb stress change caused by the Sand Point earthquake at the hypocenter of the Chignik earthquake is merely around 0.01 bars,suggesting the negligible triggering effect on the latter earthquake;(2)The triggering effects of the Simeonof,Sand Point,and Chignik earthquakes on aftershocks within three months are not well pronounced because of the triggering rates of 38%,14%,and 43%respectively.Other factors may have played an important role in promoting the occurrence of these aftershocks,such as the roughness of the subduction interface,the complicated velocity structure of the lithosphere,and the heterogeneous prestress therein;(3)The three earthquakes caused remarkable coseismic Coulomb stress changes at the subduction interface nearby these mainshocks,with an average Coulomb stress change of 3.2 bars in the shallow region directly inwards the trench.

Exercise-induced modulation of miR-149-5p and MMP9 in LPS-triggered diabetic myoblast ER stress: licorice glycoside E as a potential therapeutic target

Background:This study explores the relationship between endoplasmic reticulum(ER)stress and diabetes,particularly focusing on the impact of physical exercise on ER stress mechanisms and identifying potential therapeutic drugs and targets for diabetes-related sepsis.The research also incorporates traditional physical therapy perspectives,emphasizing the genomic insights gained from exercise therapy in disease management and prevention.Methods:Gene analysis was conducted on the GSE168796 and GSE94717 datasets to identify ER stress-related genes.Gene interactions and immune cell correlations were mapped using GeneCard and STRING databases.A screening of 2,456 compounds from the TCMSP database was performed to identify potential therapeutic agents,with a focus on their docking potential.Techniques such as luciferase reporter gene assay and RNA interference were used to examine the interactions between microRNA-149-5p and MMP9.Results:The study identified 2,006 differentially expressed genes and 616 miRNAs.Key genes like MMP9,TNF-α,and IL1B were linked to an immunosuppressive state.Licorice glycoside E demonstrated high affinity for MMP9,suggesting its potential effectiveness in treating diabetes.The constructed miRNA network highlighted the regulatory roles of MMP9,IL1B,IFNG,and TNF-α.Experimental evidence confirmed the binding of microRNA-149-5p to MMP9,impacting apoptosis in diabetic cells.Conclusion:The findings highlight the regulatory role of microRNA-149-5p in managing MMP9,a crucial gene in diabetes pathophysiology.Licorice glycoside E emerges as a promising treatment option for diabetes,especially targeting MMP9 affected by ER stress.The study also underscores the significance of physical exercise in modulating ER stress pathways in diabetes management,bridging traditional physical therapy and modern scientific understanding.Our study has limitations.It focuses on the microRNA-149-5p-MMP9 network in sepsis,using cell-based methods without animal or clinical trials.Despite strong in vitro findings,in vivo studies are needed to confirm licorice glycoside E’s therapeutic potential and understand the microRNA-149-5p-MMP9 dynamics in real conditions.

Exosomes derived from microglia overexpressing miR-124-3p alleviate neuronal endoplasmic reticulum stress damage after repetitive mild traumatic brain injury

We previously reported that miR-124-3p is markedly upregulated in microglia-derived exosomes following repetitive mild traumatic brain injury.However,its impact on neuronal endoplasmic reticulum stress following repetitive mild traumatic brain injury remains unclear.In this study,we first used an HT22 scratch injury model to mimic traumatic brain injury,then co-cultured the HT22 cells with BV2 microglia expressing high levels of miR-124-3p.We found that exosomes containing high levels of miR-124-3p attenuated apoptosis and endoplasmic reticulum stress.Furthermore,luciferase reporter assay analysis confirmed that miR-124-3p bound specifically to the endoplasmic reticulum stress-related protein IRE1α,while an IRE1αfunctional salvage experiment confirmed that miR-124-3p targeted IRE1αand reduced its expression,thereby inhibiting endoplasmic reticulum stress in injured neurons.Finally,we delivered microglia-derived exosomes containing miR-124-3p intranasally to a mouse model of repetitive mild traumatic brain injury and found that endoplasmic reticulum stress and apoptosis levels in hippocampal neurons were significantly reduced.These findings suggest that,after repetitive mild traumatic brain injury,miR-124-3 can be transferred from microglia-derived exosomes to injured neurons,where it exerts a neuroprotective effect by inhibiting endoplasmic reticulum stress.Therefore,microglia-derived exosomes containing miR-124-3p may represent a novel therapeutic strategy for repetitive mild traumatic brain injury.

Transcriptome Profiling of the Salt-Stress Response in Paper Mulberry

Paper mulberry is a high-quality woody feed resource plant with high crude protein content.It is widely distributed in China and has excellent characteristics of salt and alkali tolerance.Paper mulberry has ecological and economic importance.Salt stress has become a critical factor with the increasing degree of soil salinity that restricts plant growth.In the saline-alkali environments,transcriptome expression is altered leading to phenotypic defects in most plants.However,the regulatory mechanism related to paper mulberry’s salt-stress(SS)response is not clearly understood.In the present study de novo transcriptomic assembly was performed,and gene expression levels were measured between different SS and natural conditions(25℃)as a control for paper mulberry plants.According to the results of our study,under NaCl stress conditions,the differential gene expression was observed in the leaves of paper mulberry compared with the control.A total of 2126 differentially expressed unigenes were observed.Among these unigenes the expression of 812 DEGs was up-regulated and the expression of 1,314 DEGs was down-regulated.Additionally,The GO and KEGG analyses regarding differentially expressed unigenes(DEUs)revealed that the observed critical transcriptomic alterations under salt stress(SS)conditions were associated with primary and secondary metabolism,photosynthesis,and plant hormone signaling pathways.Further investigations such as gene function studies regarding the unigenes depicting altered expression under salt stress conditions in paper mulberry will help understand the mechanism of salt tolerance,and this information can be utilized in paper mulberry breeding and improvement programs.

Proteome-wide identification of S-sulfenylated cysteines response to salt stress in Brassica napus root

Reactive oxygen species(ROS)play a key role in a variety of biological processes,such as the perception of abiotic stress,the integration of different environmental signals,and the activation of stress response networks.Salt stress could induce an increased ROS accumulation in plants,disrupting intracellular redox homeostasis,leading to posttranslational modifications(PTMs)of specific proteins,and eventually causing adaptive changes in metabolism.Here,we performed an iodoTMT-based proteomic approach to identify the sulfenylated proteins in B.napus root responsing to salt stress.Totally,1348 sulfenylated sites in 751 proteins were identified and these proteins were widely existed in different cell compartments and processes.Our study revealed that proteins with changed abundance and sulfenylation level in B.napus root under salt stress were mainly enriched in the biological processes of ion binding,glycolysis,ATP binding,and oxidative stress response.This study displays a landscape of sulfenylated proteins response to salt stress in B.napus root and provides some theoretical support for further understanding of the molecular mechanisms of redox regulation under salt stress in plants.

Identification of novel salt stress-responsive microRNAs through sequencing and bioinformatic analysis in a unique halophilic Dunaliella salina strain

Dunaliella salina is a classic halophilic alga.However,its molecular mechanisms in response to high salinity at the post transcriptional level remain unknown.A unique halophilic alga strain,DS-CN1,was screened from four D.salina strains via cell biological,physiological,and biochemical methods.High-throughput sequencing of small RNAs(sRNAs)of DS-CN1 in culture medium containing 3.42-mol/L NaCl(SS group)or 0.05-mol/L NaCl(CO group)was performed on the BGISEQ-500 platform.The annotation and sequences of D.salina sRNAs were profiled.Altogether,44 novel salt stress-responsive microRNAs(miRNAs)with a relatively high C content,with the majority of them being 24 nt in length,were identified and characterized in DS-CN1.Twenty-one differentially expressed miRNAs(DEMs)in SS and CO were screened via bioinformatic analysis.A total of 319 putative salt stress-related genes targeted(104 overlapping genes)by novel miRNAs in this alga were screened based on our previous transcriptome sequencing research.Furthermore,these target genes were classified and enriched by GO and KEGG pathway analysis.Moreover,5 novel DEMs(dsa-mir3,dsa-mir16,dsa-mir17,and dsa-mir26 were significantly upregulated,and dsa-mir40 was significantly downregulated)and their corresponding 10 target genes involved in the 6 significantly enriched metabolic pathways were verified by quantitative real-time PCR.Next,their regulatory relationships were comprehensively analyzed.Lastly,a unique salt stress response metabolic network was constructed based on the novel DEM-target gene pairs.Taken together,our results suggest that 44 novel salt stress-responsive microRNAs were identified,and 4 of them might play important roles in D.salina upon salinity stress and contribute to clarify its distinctive halophilic feature.Our study will shed light on the regulatory mechanisms of salt stress responses.

工业大麻ZF-HD转录因子的鉴定及表达分析

【目的】对工业大麻ZF-HD(Zinc finger-homeodomain)转录因子进行鉴定和表达模式分析,为ZF-HD功能研究及工业大麻遗传性状的改良提供理论参考。【方法】对5个工业大麻ZF-HD转录因子的基因结构、蛋白理化性质、蛋白系统进化关系和启动子顺式作用元件进行生物信息学分析,通过实时荧光定量PCR(qRT-PCR)对基因表达量进行检测。【结果】5个CsZF-HD基因分布于大麻基因组的2条染色体上,其编码序列长度为783~1185 bp,编码260~394个氨基酸残基,蛋白分子量为27.95~43.33 ku,理论等电点为7.46~8.87。它们编码的蛋白均含有1个保守的ZF-HD_dimer结构域,均为不稳定的亲水性蛋白且定位于细胞核中。ZF-HD蛋白家族分为3个亚族(A、B和C),CsZF-HD1和CsZF-HD2属于A亚族,CsZF-HD3属于C亚族,CsZF-HD4和CsZF-HD5属于B亚族。CsZF-HD1和CsZF-HD2在茎和叶中的表达量相差不明显,在根中表达量最低;CsZF-HD3在茎中的表达量最高,叶中表达量最低;CsZF-HD4在叶中的表达量最高,根中表达量最低;CsZF-HD5在根中表达量最高,茎中表达量最低。5个CsZF-HD基因均对盐和干旱胁迫响应,其中CsZF-HD1和CsZF-HD3在盐和干旱胁迫下表达量升高最明显。5个CsZF-HD基因启动子序列中均含有不同种类和数量的逆境相关顺式作用元件。【结论】ZF-HD转录因子可能在工业大麻应对盐和干旱胁迫时发挥转录调控作用。

叶面喷施6-BA对盐胁迫下不同种源黑果枸杞幼苗生理指标和光合特性的影响

以青海黑果枸杞和甘肃黑果枸杞实生苗为试验材料,分析不同浓度6-苄氨基腺嘌呤(6-BA)对盐胁迫下不同种源黑果枸杞幼苗生理指标、抗氧化系统和光合参数的影响。结果表明:(1)施加低、中浓度6-BA降低了无盐条件下两种黑果枸杞叶片和根中Na^(+)和Cl^(-)质量分数、过氧化氢(H_(2)O_(2))质量摩尔浓度、电解质外渗率,提高了抗坏血酸过氧化酶(APX)、谷胱甘肽还原酶(GR)、硝酸还原酶(NR)和ATP硫酸化酶(ATPS)活性,以及还原型谷胱甘肽(GSH)、氧化型谷胱甘肽(GSSG)、氮(N)、硫(S)质量分数和光合氮利用效率;高浓度6-BA对无盐条件下两种黑果枸杞上述指标却有相反的作用。(2)与青海黑果枸杞相比,甘肃黑果枸杞在盐胁迫下叶片中积累了更多的Na^(+)和Cl^(-),并遭受更大的氧化胁迫,表现为叶片中更高的H_(2)O_(2)质量摩尔浓度和电解质渗出率以及更低的光合参数。(3)受到盐胁迫后,青海黑果枸杞体内抗坏血酸过氧化物酶(APX)和谷胱甘肽还原酶(GR)活性高于甘肃黑果枸杞,但甘肃黑果枸杞的超氧化物歧化酶(SOD)活性更高。(4)与甘肃黑果枸杞相比,青海黑果枸杞叶中的氮和硫通过提高同化酶—硝酸还原酶(NR)和ATP硫酸化酶(ATPS)活性而积累更多,从而使叶片中还原型谷胱甘肽(GSH)质量分数显著提高。(5)低、中浓度6-BA有利于提高青海黑果枸杞和甘肃黑果枸杞氮硫同化效率、GSH质量分数以及APX和GR的活性,导致在无盐条件下光合作用增强,缓解了盐胁迫下光合作用的下降,限制了叶片中Na^(+)和Cl^(-)的质量分数,提高了光合氮利用效率;然而高浓度6-BA的应用对黑果枸杞的生长具有反作用;6-BA对青海黑果枸杞盐胁迫下各生理指标变化的影响比甘肃黑果枸杞更明显。因此,6-BA的施用,使青海黑果枸杞的NR和ATPS活性提高,增强了抗氧化防御系统的功能,缓解了盐胁迫引起光合作用减弱的现象。

王族海棠Alfin-like家族基因的鉴定及盐胁迫下的表达分析

Alfin-like(AL)转录因子家族对非生物胁迫反应具有重要的调控作用。本研究采用同源比对的方法检索鉴定王族海棠AL转录因子家族基因,系统分析其生物信息学特性,并采用qRT-PCR方法分析其在盐胁迫下的表达模式,以期为揭示AL家族在王族海棠响应盐胁迫中的生理功能提供理论依据。结果表明,从王族海棠基因组中共鉴定出11个MdALs基因,分布在6条染色体上,分别命名为MdAL1—MdAL11。MdALs转录因子具有高度保守的Alfin结构域和PHD锌指结构域,含4~10个内含子;上游启动子区域有大量与植物激素和非生物胁迫响应相关的顺式作用元件,且基因片段复制事件在MdAL基因家族的扩展中起着至关重要的作用。转录组分析显示MdALs基因主要在王族海棠生育后期高表达。qRT-PCR分析进一步表明,MdALs基因在王族海棠遭受盐胁迫下的表达模式不同,盐胁迫下,MdAL3、MdAL4、MdAL6基因的表达量整体上调,尤其MdAL4基因,其表达水平随着盐胁迫处理时间的延长呈显著增加趋势。综上,王族海棠AL转录因子家族与植物响应激素变化和非生物胁迫密切相关,尤其是MdAL4基因,强烈响应盐胁迫,这可为利用基因工程技术改良王族海棠种质奠定基础。

高耐盐紫球藻资源筛选及其miR398-CSD盐胁迫应答模式研究

紫球藻是一类极具开发潜力的特色微藻,本研究旨在筛选、鉴定一批高耐盐紫球藻株,并对其miR398-CSD盐胁迫应答模式进行研究,以期为特色紫球藻的工业化高效应用及其耐盐机理机制的研究奠定基础。以国内外不同品系的紫球藻为试材,采用形态学、生理学、系统进化的方法分析其基本特征,从中筛选高耐盐品系;用高通量测序及生物信息学技术,鉴定紫球藻miR398及其靶基因CSD序列,分子生物学技术对其抑制类型进行验证;用stem-loop qRT-PCR和qRT-PCR技术,对紫球藻高耐盐品系不同盐度胁迫下的miR398及CSD表达模式进行分析。7株藻株均属于紫球藻属,系统进化上可划分为3簇,其细胞形态均符合典型的紫球藻特征,P1和P4属于高耐盐品系;miR398以切割抑制的方式对其靶基因CSD进行抑制,紫球藻miR398的表达随盐度的升高而下调,CSD则随盐度的升高而上调。研究结果为高耐盐紫球藻资源的开发利用及miRNAs分子定向改良藻株品质奠定基础。

外源γ-氨基丁酸缓解燕麦幼苗盐碱胁迫的生理效应

为探究外源γ-氨基丁酸(GABA)对燕麦耐盐碱性的影响,以燕麦白燕2号为试验材料,在混合盐碱(NaCl∶Na_(2)SO_(4)∶NaHCO_(3)∶Na_(2)CO_(3)摩尔比为1∶9∶9∶1)胁迫下叶面喷施不同浓度GABA,测定燕麦幼苗叶绿素含量、荧光参数、渗透调节物质及抗氧化酶活性的变化,同时对幼苗生长缓解效应进行综合评价。结果表明,外源喷施4~6 mmol·L^(-1) GABA可显著提高盐碱胁迫下燕麦幼苗的叶绿素含量和光系统Ⅱ反应活性,促进光合作用;在盐碱胁迫下,与喷施清水相比,燕麦幼苗叶面喷施不同浓度GABA后,脯氨酸(Pro)、丙二醛(MDA)和可溶性糖(SS)含量分别下降了14.7%~29.7%、28.2%~54.4%和1.8%~11.8%,超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)活性分别提高了17.9%~58.3%、4.4%~33.4%和8.3%~19.1%。经隶属函数法综合评价得出,叶面喷施GABA提高燕麦幼苗耐盐碱性的最佳浓度为6 mmol·L^(-1)。以上结果说明,叶面喷施适宜浓度的GABA能够提高燕麦幼苗光合能力和抗氧化酶活性,降低渗透调节物质含量,提高燕麦幼苗的抗盐碱能力,有效缓解盐碱胁迫对幼苗生长带来的伤害。

Cp2-EPS对盐胁迫下紫花苜蓿苗期形态及光合特性的影响

胞外多糖(Exopolysaccharides, EPS)能提高植物对非生物胁迫的抵抗力。桃色欧文氏菌(Erwinia persicina)Cp2高产EPS,但其对紫花苜蓿(Medicago sativa)耐盐性的作用不明。因此,本试验以紫花苜蓿‘巨能551’为材料进行沙培试验,探究不同浓度(0,0.5,1.0,1.5和2.0 g·L^(-1))Cp2-EPS对100 mmol·L^(-1)NaCl胁迫下苜蓿苗期盐害的减缓效果。结果表明:100 mmol·L^(-1)NaCl胁迫可显著抑制紫花苜蓿的生长,而添加Cp2-EPS后则对苜蓿生长表现出低浓度促进高浓度抑制的现象。当Cp2-EPS浓度为0.5~1.5 g·L^(-1)时,可显著促进苜蓿植株的生长,显著提高苜蓿幼苗的鲜干重、相对含水量、叶绿素含量和净光合速率并降低叶片相对电导率,表明0.5~1.5 g·L^(-1)Cp2-EPS能够缓解该胁迫对苜蓿的毒害;而当Cp2-EPS浓度达到2.0 g·L^(-1)时则加剧了该胁迫对苜蓿的毒害。最后,综合评价结果表明:1.0 g·L^(-1)Cp2-EPS对该盐浓度下紫花苜蓿的缓解效果最佳。本研究为开发利用改良盐碱地种植紫花苜蓿的生物源制剂奠定了实践和理论基础。

Functional analysis of flavanone 3-hydroxylase(F3H)from Dendrobium officinale,which confers abiotic stress tolerance

Flavonoids are important bioactive components in Dendrobium officinale,a medicinal orchid.They are involved in many biological activities,including protecting plants against biotic and abiotic stresses.Research on the key genes related to flavonoid biosynthesis in D.officinale is limited.In this study,one of the key flavonoid biosynthesis genes,flavanone 3-hydroxylase(F3H),was characterized from D.officinale.The open reading frame of DoF3H was 1134 bp long and it encoded a 377-amino acid protein.The DoF3H protein showed considerably high homology with F3H proteins from other plant species and shared a common evolutionary ancestor with other F3Hs.DoF3H transcripts were detected in different organs of adult plants and mainly accumulated in flowers,followed by roots,stems and leaves,a pattern that was similar to the content of flavonoids.Recombinant DoF3H protein,which was localized in the cytosol,could convert naringenin to dihydrokaempferol.The mRNA levels of DoF3H were significantly induced by salt and cold stresses.Furthermore,the heterologous expression of DoF3H in Escherichia coli conferred it higher tolerance to salt and cold stresses.These results provide insight into the molecular function of DoF3H in the biosynthesis of flavonoids,and provide a new application for improvement of abiotic tolerance in D.officinale.

Effects of Different Arbuscular Mycorrhizal Fungi on Physiology of Viola prionantha under Salt Stress

Arbuscular mycorrhizal(AM)fungi distribute widely in natural habits and play a variety of ecological functions.In order to test the physiological response to salt stress mediated by different AM fungi,Viola prionantha was selected as the host,the dominant AM fungus in the rhizosphere of V.philippica growing in Songnen saline-alkali grassland,Rhizophagus irregularis,and their mixtures were used as inoculants,and NaCl stress was applied after the roots were colonized.The results showed that V.philippica could be colonized by AM fungi in the field and the colonization rate ranged from 73.33%to 96.67%,and Claroideoglomus etunicatum was identified as the dominant AM fungi species in the rhizosphere of V.philippica by morphology combined with sequencing for AM fungal AML1/AML2 target.Inoculation with both the species resulted in the formation of mycorrhizal symbiosis(the colonization rate was more than 70%)and AM fungi significantly enhanced plants’tolerance to salt stress of varying magnitude.Higher activity of antioxidant enzymes and augmented levels of proline and other osmoregulators were observed in AM plants.The content of MDA in CK was higher than that in the inoculations with the stress of 100,200,and 250 mM.All indices except soluble protein content and MDA content were significantly correlated with AM fungal colonization indices.The analysis for different AM fungal effects showed that the mixtures and R.irregularis worked even better than C.etunicatum.These results will provide theoretical support for the exploration and screening of salt-tolerant AM fungi species and also for the application of AM-ornamental plants in saline-alkali urban greening.

基于NOX4/ROS/NF-κB通路探讨补阳还五汤对Dahl盐敏感性大鼠高血压肾损伤的保护机制

目的通过观察研究补阳还五汤对Dahl盐敏感大鼠烟酰胺腺嘌呤二核苷酸磷酸氧化酶4(NOX4)/活性氧/促使核转录因子-кB(NOX4/ROS/NF-κB)信号转导通路的影响,探讨补阳还五汤对Dahl盐敏感性大鼠高血压肾损伤的保护机制。方法7周龄Dahl盐敏感大鼠60只,分为正常组,模型组,补阳还五汤低、中、高剂量组和氢氯噻嗪共6组,每组10只。所有大鼠于SPF级动物实验中心适应性饲养7 d,均给予普通饲料。随后进入实验阶段,正常组每日采用低盐饲料喂养(NaCl含量0.3%饲料),其余各组采用高盐饲料喂养(NaCl含量8%),7 d后开始药物干预,补阳还五汤低、中、高剂量组按成人和大鼠给药量换算,每100 g大鼠每日给药量分别为0.275、0.55、1.1 g,氢氯噻嗪组每100 g大鼠每日给药量为1 mg。造模期间,每隔7 d定时检测大鼠收缩压。高盐饲养35 d后检测到模型组大鼠收缩压显著升高,病理组织学观察发现模型组大鼠肾脏损伤明显,则说明Dahl盐敏感性大鼠高血压肾损伤模型造模成功。随后取各组大鼠肾脏进行检测,采用酶联免疫吸附法(enzyme linked immunosorbent assay,ELISA)检测大鼠肾脏中过氧化氢酶(catalase,CAT)、超氧化物歧化酶(superoxide dismutase,SOD)、丙二醛(malondialdehyde,MDA)以及ROS表达含量;HE染色观察大鼠肾脏病理学变化;荧光定量PCR检测大鼠肾脏中NOX4、p47phox、NF-кB、白介素-6(interleukin-6,IL-6)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)的mRNA表达,Western blot检测大鼠肾脏中NOX4、p47phox、NF-κB、IL-6、TNF-α蛋白表达。结果与模型组大鼠相比补阳还五汤中、高剂量组大鼠收缩压下降明显(P<0.05);肾脏中CAT、SOD含量显著升高(P<0.05),MDA、ROS显著降低(P<0.05);HE染色结果显示,补阳还五汤中、高剂量组大鼠肾组织形态较模型组明显改善;NOX4、p47phox、NF-κB、IL-6、TNF-αmRNA表达和蛋白表达均降低(P<0.05)。结论补阳还五汤可能是通过调节NOX4/ROS/NF-κB通路相关信号分子的表达,抑制氧化应激与炎症反应,从而起到保护Dahl盐敏感性大鼠高血压肾损伤的作用。

水淹-盐胁迫对两耐淹树种生理生态及Na^(+)、K^(+)和Cl^(-)离子积累的影响

滨海河口河岸带植物易受到水淹和盐复合胁迫的影响。前期研究表明水翁(Syzygium nervosum A.Cunn.ex DC.)和乌墨(Syzygium cumini(L.)Skeels)具有较强的耐水淹能力,然而两种植物对水淹-盐复合胁迫的耐受性尚不清楚。本研究设置水淹、盐(350 mmol/L)及水淹-盐(175 mmol/L)复合胁迫3种胁迫处理,比较两物种对不同胁迫的生长、生理生化和盐胁迫相关离子积累的响应。结果显示,经过24 d处理,3种胁迫处理均低了两物种的生物量,提高了还原型谷胱甘肽(GSH)含量、抗坏血酸过氧化物酶(APX)和谷胱甘肽还原酶(GR)的活性。水淹胁迫下两物种的超氧阴离子(O_(2)^(·-))和过氧化氢(H_(2)O_(2))含量下降,根系K^(+)含量上升,K^(+)/Na^(+)下降;盐胁迫下乌墨幼苗O_(2)^(·-)含量下降,水翁幼苗O_(2)^(·-)含量上升,两物种幼苗的脯氨酸含量、H_(2)O_(2)含量、Na^(+)、Cl^(-)含量上升,K^(+)/Na^(+)下降;水淹-盐复合胁迫下两物种的O_(2)^(·-)、Na^(+)、Cl^(-)含量上升,K^(+)/Na^(+)下降。此外,与水淹相比,水淹-盐复合胁迫降低了两物种的生物量,提升了两物种的O_(2)^(·-)、Na^(+)、Cl^(-)含量。与水翁相比,乌墨幼苗在盐、水淹-盐复合胁迫下有较高的生物量、脯氨酸含量、GR活性和K^(+)/Na^(+),以及较低的H_(2)O_(2)、Na^(+)和Cl^(-)含量,具有更强的盐、水淹-盐复合胁迫耐受能力。

Analysis of Organic Acids Accumulated in Kochia Scoparia Shoots and Roots by Reverse-phase High Performance Liquid Chromatography Under Salt and Alkali Stress

Several organic acids accumulated in Kochia Scoparia shoots and roots were studied by means of reverse-phase high performance liquid chromatography with a C18 column. Five types of binary organic acids were separated. The organic acid concentrations were determined in K. Scoparia seedlings stressed by saline （NaCI） and alkaline （NaHCO3） at the same Na^＋ concentration. Concentrations of organic acids are stimulated by alkaline because the cells will adjust their pH values through the accumulation of organic acids, when the environment is basic. The concentrations of oxalic acid and succinic acid are higher than those of other organic acids, including tartaric acid and malic acid, and the concentration of citric acid is the lowest. The concentrations of the organic acids in the roots are higher than those in the shoots under salt（NaCI） stress, but the results are opposite while the roots are under alkali （ NaHCO3 ） stress. This indicates that there are different adaptive strategies for K. Scopar/a seedlings in organic acid metabolism under salt and alkali stress.