PEG胁迫对香石竹幼苗的影响及香石竹抗旱性评价

为探究干旱胁迫对香石竹幼苗的影响,鉴定香石竹各品种的抗旱能力,以9个香石竹品种为研究对象,采用PEG 6000模拟干旱胁迫,分析叶片相对含水量(RWC)、超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)、可溶性蛋白、丙二醛(MDA)、脯氨酸(Pro)等7个生理生化指标的变化,并基于这7个理化指标的抗旱指数,对香石竹各品种进行主成分分析及综合评价。结果表明:9个香石竹品种在水分胁迫处理时各品种各指标存在显著差异(P0.05)。将7个抗旱指标用主成分分析法归纳成3个主成分,累积贡献率为84.46%,利用隶属函数法评定的抗旱性强弱顺序为马斯特、苔丝、皇族、紫罗兰、火焰、粉恋、锦葵钛合金、粉黛、自由。     

利用隶属函数法对4种珍珠菜属过路黄抗旱性的综合评价

在干旱胁迫条件下,测定4种过路黄的细胞质外渗率、MDA含量、SOD活性、POD活性、脯氨酸、可溶性糖、可溶性蛋白质含量等理化指标,利用隶属函数法对材料进行抗旱性综合评价。通过综合隶属函数值评定的抗旱性强弱顺序为：广西过路黄>花叶过路黄、金叶过路黄>聚花过路黄。同时表明,抗旱性是一个综合指标。     

OsMas1, a novel maspardin protein gene,confers tolerance to salt and drought stresses by regulating ABA signaling in rice

Drought and salt stresses, the major environmental abiotic stresses in agriculture worldwide, affect plant growth, crop productivity, and quality.  Therefore, developing crops with higher drought and salt tolerance is highly desirable.  This study reported the isolation, biological function, and molecular characterization of a novel maspardin gene, OsMas1, from rice.  The OsMas1 protein was localized to the cytoplasm.  The expression levels of OsMas1 were up-regulated under mannitol, PEG6000, NaCl, and abscisic acid (ABA) treatments in rice.  The OsMas1 gene was introduced into the rice cultivar Zhonghua 11 (wild type, WT).  OsMas1-overexpression (OsMas1-OE) plants exhibited significantly enhanced salt and drought tolerance; in contrast, OsMas1-interference (OsMas1-RNAi) plants exhibited decreased tolerance to salt and drought stresses, compared with WT.  OsMas1-OE plants exhibited enhanced hypersensitivity, while OsMas1-RNAi plants showed less sensitivity to exogenous ABA treatment at both germination and post-germination stages.  ABA, proline and K⁺ contents and superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), and photosynthesis activities were significantly increased.  In contrast, malonaldehyde (MDA), hydrogen peroxide (H₂O₂), superoxide anion radical (O₂^-·), and Na⁺ contents were significantly decreased in OsMas1-OE plants compared with OsMas1-RNAi and WT plants.  Overexpression of OsMas1 up-regulated the genes involved in ABA signaling, proline biosynthesis, reactive oxygen species (ROS)-scavenging system, photosynthesis, and ion transport under salt and drought stresses.  Our results indicate that the OsMas1 gene improves salt and drought tolerance in rice, which may serve as a candidate gene for enhancing crop resistance to abiotic stresses.     

甘蔗新品系对干旱胁迫的生理响应及抗旱性评价

【目的】探究甘蔗10个优良新品系的抗旱性能,为抗旱品种选育及推广应用提供科学依据。【方法】在人工干旱胁迫和正常供水处理下测定甘蔗伸长初期的根系活力,叶片脯氨酸、可溶性蛋白、可溶性糖、丙二醛(MDA)含量,叶片超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)活性8项生理指标的变化;采用主成分分析与聚类分析方法综合评价甘蔗伸长初期的抗旱性。【结果】受干旱胁迫后,甘蔗根系活力降低,叶片可溶性蛋白、可溶性糖、脯氨酸、MDA含量升高,叶片SOD、POD、CAT活性增强。聚类分析将包括对照品种ROC22在内的11个甘蔗品系分成抗旱性强、中度抗旱和抗旱性弱3类。抗旱性强的新品系为A6-13115和A3-1320,中度抗旱的新品系为A13-1396和A6-13122,抗旱性弱的新品系为A6-13111、A11-1390、A1-1305、A7-13120、A7-13104和A4-1316。【结论】采用主成分和聚类分析方法对多个指标综合评价能提高甘蔗抗旱评价的可靠性、准确性。甘蔗新品系A6-13115和A3-13120抗旱性强。     

Genome-wide identification and analysis of the regulation wheat DnaJ family genes following wheat yellow mosaic virus infection

The co-chaperone DnaJ plays an important role in protein folding and regulation of various physiological activities, and participates in several pathological processes. DnaJ has been extensively studied in many species including humans, drosophila, mushrooms, tomatoes, and Arabidopsis. However, few studies have examined the role of DnaJ in wheat (Triticum aestivum), and the interaction mechanism between TaDnaJs and plant viruses. Here, we identified 236 TaDnaJs and performed a comprehensive genome-wide analysis of conserved domains, gene structure and protein motifs, chromosomal positions and duplication relationships, and cis-acting elements. We grouped these TaDnaJs according to their domains, and randomly selected six genes from the groups for tissue-specific analysis, and expression profiles analysis under hormone stress, and 17 genes for plant virus infection stress. In qRT-PCR, we found that among the 17 TaDnaJ genes tested, 16 genes were up-regulated after wheat yellow mosaic virus (WYMV) infection, indicating that the TaDnaJ family is involved in plant defense response. Subsequent yeast two-hybrid assays verified the WYMV NIa, NIb and 7KD proteins interacted with TaDJC (TraesCS7A02G506000), which had the most significant changes in gene expression levels after WYMV infection. Insights into the molecular mechanisms of TaDnaJ-mediated stress tolerance and sensitivity could inform different strategies designed to improve crop resistance to abiotic and biotic stress. This study provides a basis for future investigation of the TaDnaJ family and plant defense mechanisms.     

‘84K’杨组氨酸激酶基因PaHK3a的表达及功能分析

  目的  干旱、高盐等逆境胁迫严重影响了植物的生长发育。研究表明,组氨酸激酶在植物逆境响应中起重要作用。本研究对银腺杨‘84K’组氨酸激酶基因PaHK3a在不同组织的表达模式分析,检测了其对生长素、细胞分裂素等植物激素处理下及人工干旱、盐碱等非生物胁迫下的表达,结合干旱、盐碱条件下丙二醛（MDA）及保护酶活性等生化指标,对该基因的功能进行了初步鉴定,为杨树抗逆分子育种研究奠定基础。  方法  以‘84K’杨无菌苗为材料,通过实时定量PCR（qRT-PCR）技术分析PaHK3a基因在不同组织的表达模式。对‘84K’杨无菌苗进行浓度为10 mmol/L植物激素处理（ABA、6-BA、IBA、GA3及水杨酸（SA））及非生物胁迫处理（42 ℃高温、0 ℃低温、200 mmol/L NaCl和5% PEG6000）,采用qRT-PCR技术分析PaHK3a基因对不同植物激素及非生物胁迫的表达响应;进一步对温室‘84K’杨进行自然干旱处理（6、8、10 d）、200 mmol/L NaCl（2、4、6 d）处理,测定不同胁迫时间点叶片PaHK3a基因的表达,以及超氧化物歧化酶（SOD）、过氧化物酶（POD）活性及MDA含量,并分析PaHK3a基因表达与生理指标的相关性,初步鉴定杨树PaHK3a基因的功能。  结果  qRT-PCR结果显示,PaHK3a基因在叶片中表达量最高,根部中等,茎段最低。与正常条件下相比,在高温、低温、NaCl及PEG模拟干旱处理时,PaHK3a基因表达量与对照相比明显增高,分别为对照的2.63、1.49、1.54、1.58倍。用IBA诱导处理时,基因表达量与对照相比差异不大,而在6-BA、ABA、GA3及SA处理时,基因表达量与对照相比均呈现显著下调。在温室干旱、盐碱胁迫处理过程中,PaHK3a基因表达均显著高于对照,呈现先上升后下降的表达模式,MDA含量也呈现类似的趋势,而SOD活性则随处理时间的延长而持续升高,POD活性在干旱胁迫时先上升后下降,而高盐胁迫时呈上升趋势。生理指标与PaHK3a基因表达量相关系分析发现,在干旱和盐胁迫下,PaHK3a基因表达量与叶片MDA含量、SOD活性和POD活性均呈正相关。  结论  PaHK3a基因在‘84K’杨根茎叶中均有表达,且叶中表达量最高;PaHK3a基因表达受细胞分裂素6-BA、GA3及ABA及SA等植物激素的负调控,同时,受温度胁迫、盐胁迫、水分胁迫等非生物胁迫正调控;温室人工干旱盐碱胁迫过程中,PaHK3a基因表达量升高,且与叶片MDA含量、SOD活性、POD活性均具有正相关性。研究结果初步显示,杨树PaHK3a基因参与杨树植物激素激素信号响应,并在抗逆境胁迫过程中发挥重要调控作用。      

白桦BpSPL8启动子的克隆及异源过表达BpSPL8对拟南芥耐旱性的影响

目的目前对植物SPL8的基因功能研究主要集中在开花和育性方面,而其在干旱胁迫响应中的作用却鲜有报道。本文克隆、分析了白桦BpSPL8启动子,并研究了BpSPL8基因在拟南芥中响应干旱胁迫的功能。方法通过PCR克隆技术得到了白桦BpSPL8启动子;利用PLACE和PlantCARE软件对BpSPL8启动子顺式作用元件进行了预测。构建了BpSPL8启动子驱动GUS（β-葡萄糖苷酸酶编码基因）的植物表达载体,并采用浸花法将其转化至拟南芥中;继而利用GUS染色分析了BpSPL8启动子的组织表达模式;同时对BpSPL8在PEG处理下的表达水平进行了qRT-PCR分析。最后,以过表达BpSPL8拟南芥为材料来探究BpSPL8在干旱胁迫下的生物学功能。结果启动子元件分析显示,BpSPL8启动子中含有组织特异表达、光响应、激素响应及多个胁迫响应元件。GUS染色结果表明,BpSPL8启动子可在拟南芥的下胚轴、叶片、叶柄、根和花序中启动GUS基因表达。BpSPL8基因在PEG处理下的野生型白桦的根和叶片中均呈现先上调后下调的表达趋势。干旱胁迫下,过表达BpSPL8拟南芥的存活率和脯氨酸含量均显著低于野生型,丙二醛含量显著高于野生型;两个已知的抗逆基因DR29B和P5CS1在干旱处理后的野生型和转基因拟南芥中均上调表达;但在转基因拟南芥中呈现出延迟上调的表达模式。结论异源过表达白桦BpSPL8能够降低拟南芥的耐旱性,并在干旱胁迫下影响抗性基因DR29B和P5CS1的表达模式。      

外源水杨酸对野生龙葵幼苗干旱胁迫的缓解效应

利用20%的PEG6000模拟干旱胁迫,同时用不同浓度（0、0.5、1.0、1.5、2.0 mmol·L^-1）的水杨酸(SA)喷施幼苗,研究了不同浓度水杨酸处理对干旱胁迫下野生龙葵幼苗生长的影响。结果表明：干旱胁迫下,龙葵幼苗植株变矮、鲜重降低、根长缩短,叶绿素和可溶性蛋白含量减少,抗氧化酶活性、游离脯氨酸和丙二醛含量增加;经SA处理干旱胁迫下龙葵幼苗株高、根长和鲜重都有明显增加,特别是1.5 mmol·L^-1 SA处理组分别增加20.8%、28.0%和29.7%,1.5 mmol·L^-1 SA处理组龙葵幼苗叶绿素含量提高31.9%、脯氨酸含量提高65.4%,可溶性蛋白含量提高42.8%,超氧化物歧化酶（SOD）、过氧化物酶（POD）与过氧化氢酶（CAT）活性达到最高,与对照相比分别增加了30.2%、87.8%和50.3%;SA处理使丙二醛（MDA）含量降低了37.8%。综合来看,适当浓度的SA可以缓解干旱对龙葵幼苗造成的伤害,并以1.5 mmol·L^-1外源SA缓解效果最好。     

拟南芥组蛋白甲基化SDG26基因突变体抗旱生理生化研究

对拟南芥SDG26基因功能缺失突变体sdg26植株和对照野生型Col植株在干旱条件下的SOD活性和MDA、可溶性糖、脯氨酸含量进行测定。结果显示:随着干旱时间(5、8、11、14、17、20 d)持续,sdg26植株中的SOD活性和MDA、可溶性糖、脯氨酸含量都表现出明显提高的趋势,上升水平显著高于对照野生型Col植株;在持续干旱20 d条件下,Col植株的叶片逐渐失水干枯及至死亡,但sdg26植株能正常生长,表现出良好的生长势,推测SDG26基因功能的丧失可以增强拟南芥的抗旱能力,说明植物的抗旱能力与组蛋白的甲基化修饰密切相关。     

Overexpression of a Cytosolic Ascorbate Peroxidase Gene,OsAPX2, Increases Salt Tolerance in Transgenic Alfalfa

Alfalfa (Medicago sativa L.) is an important forage crop in the world and it is of great significance for the improvement of its salt tolerance. To improve salt tolerance in alfalfa, a rice ascorbate peroxidase gene (OsAPX2) was introduced into alfalfa using Agrobacterium tumefaciens-mediated transformation with marker gene bar. The different T-DNA insertions in T₁ transgenic alfalfa were identified by Southern hybridization. Three independent T₂ transgenic lines were selected for stress analysis and the results showed that all of them were salt tolerant compared with wild-type plants. The transgenic plants had low levels of H₂O₂, malondialdehyde and relative electrical conductivity under salt and drought stresses. Moreover, the contents of chlorophyll and proline, and APX activity were high in transgenic plants under salt and drought stresses. Taken together, the overexpression of OsAPX2 enhances salt tolerance in alfalfa through scavenging reactive oxygen species.     

Melatonin treatment alleviates chilling injury in mango fruit 'Keitt' by modulating proline metabolism under chilling stress

Mangoes often suffer from low temperature-induced chilling injury (CI) during postharvest cold storage.  Therefore, advanced techniques are crucial and in high demand to solve the chilling stress of mango fruit for a higher value.  This study addresses chilling stress modulation by investigating the effects of melatonin treatment on CI, proline metabolism, and related gene expressions of ‘Keitt’ mango during cold storage after dipped in 0 (control), 0.1 (MT1), and 0.2 mmol L^–1 (MT2) melatonin solution for 30 min.  The results revealed that melatonin treatment in MT1 significantly reduced CI development and increased proline content in mango fruit during cold storage compared to the control.  These changes were along with increases in the activity of critical enzymes as well as the expression of encoding genes involved in proline biosynthesis, such as pyrroline-5-carboxylate synthetase (P5CS), pyrroline-5-carboxylate reductase (P5CR), ornithine D-aminotransferase (OAT), P5CS2, P5CR2, and OAT3.  Additionally, proline dehydrogenase (PDH) activity and the expression of the PDH3 gene associated with proline dehydrogenation were lower in MT1-treated mangoes than the controlled group.  Thus, melatonin treatment has regulated proline metabolism resulting in the accumulation of proline, subsequently contributing to enhancing the chilling tolerance of ‘Keitt’ mango fruit.     

茶氨酸诱导烟草幼苗抵御干旱胁迫的研究

通过施用茶氨酸来减轻干旱胁迫对烟草幼苗的毒害,并从抗氧化物质和抗氧化酶来探讨其作用机理。结果表明,在干旱胁迫下,0.25 mmol·L-1茶氨酸能显著改善烟草幼苗的生长情况,提高其存活率;外源茶氨酸预处理不仅能显著提高干旱胁迫下烟草幼苗谷胱甘肽(GSH)含量,还参与调节烟草幼苗超氧化物酶(SOD)过氧化物酶(POD)、过氧化氢酶(CAT)和抗坏血酸还原酶(APX)活性,降低相对电导率、脯氨酸、过氧化氢(H2O2)和丙二醛(MDA)含量。茶氨酸诱导烟草幼苗抵御干旱胁迫是通过激发植物体抗氧化物质及抗氧化酶活性调节的综合结果。     

共转化法获得蒙农杂种冰草转基因植株(征文)

本研究以冰草属（Agropyron Gaertn.）中的一个优质种间杂种——“蒙农杂种”冰草（A. cristatum хA.desertorum cv.‘Hycrest-Mengnong’ ）为材料，在以幼穗为外植体建立的冰草组织培养再生体系基础上，以调控脯氨酸生物合成最后一步的关键酶的突变体基因p5CS为目标基因，bar基因为筛选标记基因，采用共转化法利用基因枪轰击冰草幼穗诱导的愈伤组织获得转基因植株,为冰草的遗传改良提供新种质。PCR和Southern检测表明外源基因p5CS已整合到冰草植物基因组DNA中；RT-PCR检测表明目的基因已在冰草转基因植株的转录水平表达；p5CS基因的遗传转化率为0.11%。     

Overexpression of GmBIN2, a soybean glycogen synthase kinase 3 gene,enhances tolerance to salt and drought in transgenic Arabidopsis and soybean hairy roots

Glycogen synthase kinase 3 (GSK3) is a kind of serine/threonine kinase widely found in eukaryotes. Many plant GSK3 kinases play important roles in regulating stress responses. This study investigated BRASSINOSTEROID-INSENSITIVE 2 (GmBIN2) gene, a member of the GSK3 protein kinase family in soybean and an orthologue of Arabidopsis BIN2/AtSK21. GmBIN2 expression was increased by salt and drought stresses, but was not significantly affected by the ABA treatment. To examine the function of GmBIN2, transgenic Arabidopsis and transgenic soybean hairy roots were generated. Overexpression of GmBIN2 in Arabidopsis resulted in increased germination rate and root length compared with wild-type plants under salt and mannitol treatments. Overexpression of GmBIN2 increased cellular Ca²⁺ content and reduced Na+ content, enhancing salt tolerance in transgenic Arabidopsis plants. In the soybean hairy root assay, overexpression of GmBIN2 in transgenic roots also showed significantly higher relative root growth rate than the control when subjected to salt and mannitol treatments. Measurement of physiological indicators, including proline content, superoxide dismutase (SOD) activity, and relative electrical conductivity, supported this conclusion. Furthermore, we also found that GmBIN2 could up-regulate the expression of some stress-related genes in transgenic Arabidopsis and soybean hairy roots. Overall, these results indicated that GmBIN2 improved tolerance to salt and drought in transgenic Arabidopsis and soybean hairy roots.     

皂角苗木对干旱胁迫的生理生化反应

对皂角Gleditsia sinensis的一年生实生苗进行盆栽控水干旱处理，并测定皂角的各个生理生化指标．结果表明：随着干旱胁迫的增强，超氧化物歧化酶（SOD）活性呈现先降后升；过氧化氢酶（CAT）活性、过氧化物酶（POD）活性、脯氨酸含量、可溶性糖含量及丙二醛（MDA）含量均表现为先升后降的趋势；苗木的可溶性蛋白含量及相对电导率随干旱胁迫强度的增加而增加．在干旱胁迫下，POD活性与CAT活性及脯氨酸含量、可溶性糖含量与脯氨酸含量的关系密切．在轻度干旱胁迫下，皂角表现出一定的抗旱性，可能与POD、CAT活性和渗透调节物质的影响有关．