中国的盐生植物

本文主要讨论了有关中国盐生植物的五个方面的问题：(1)中国盐生植物的种类;(2)中国盐生植物的类型;(3)中国盐生植物的植被类型;(4)世界盐生植物名录中漏录的中国盐生植物;(5)中国盐生植物的经济潜势。最后还讨论了我国今后盐生植物研究方向和重点。     

中国的盐生植物

本文主要讨论了有关中国盐生植物的五个方面的问题：（１）中国盐生植物的种类；（２）中国盐生植物的类型；（３）中国盐生植物的植被类型；（４）世界盐生植物名录中漏录的中国盐生植物；（５）中国盐生植物的经济潜势。最后还讨论了我国今后盐生植物研究方向和重点。     

盐害生理与植物抗盐性

概述了盐害对植物的伤害及植物耐盐的生理机制,并综述了植物耐盐相关基因的研究进展。同时综合相关资料,提出了提高植物耐盐性的途径。     

植物耐盐研究进展

综述了盐胁迫对植物的损伤和其中的各种生理生化过程,以及植物在抵抗盐胁迫过程中的耐盐机理。新的研究成果表明,植物自身的miRNA可能在植物抗逆境过程中起到了重要作用,甲基化过程参与了抗逆境相关的甜菜碱等小分子有机物质的合成。     

植物的抗盐生理和盐害的防治

茫茫海边,见不到绿油油的庄稼,但仍可见到有些植物健壮地生长着。那么,这些植物是怎样适应盐碱严重的土壤环境的呢？科学家们经过研究发现,多数植物之所以不能在盐碱地上正常生长,是因为盐分对植物有很大危害。这些危害主要包括：（1）盐分对植物细胞产生的直接伤害,尤其是对膜系统,使膜的组分、透性、运输等都发生变化,影响正常生理功能。（2）影响细胞对其它离子的吸收,破坏细胞内在环境的离子平衡,导致代谢紊乱。（3）土壤盐分过多,导致土壤水势下降,对植物产生渗透胁迫,影响植物水分吸收,甚至引起细胞脱水,膨压降低,使…     

植物盐胁迫蛋白

本文概述了植物盐胁迫蛋白(saltstress proteins)的种类、性质、分布和可能的生理意义以及与其他逆境蛋白的联系,并介绍对植物耐盐性分子基础的探索。     

植物耐盐蛋白的研究

植物耐盐蛋白的研究邵宏波,初立业（四平师范学院生物工程研究所,吉林四平１３６０００）关键词植物耐盐蛋白迄今为止,世界上还没有真正育成一种耐盐的作物品种。究其原因,就是不了解植物耐盐的分子生物学基础。近年来,有关植物在盐胁迫条件下基因表达变化的研究引起...     

盐生植物盐爪爪的耐盐生理特性探讨

当前土壤盐渍化日益严重,是限制植物生长的一个主要环境因子,然而在盐碱自然环境中生长着许多耐盐植物,为更好地了解盐生植物的耐盐机理,该文从无机离子Na+,K+,Ca2+含量、脯氨酸水平、水势变化、丙二醛含量和盐胁迫的表型等生理参数以及半定量RT-PCR检测脯氨酸合成关键酶基因(P5CS)的表达规律等方面探讨盐胁迫下盐爪爪的耐盐特性。结果表明:(1)随着盐浓度的升高,Na+在根和肉质化的叶中显著地富集,且叶中积累的Na+比根中更多;(2)在盐胁迫条件下,随着盐浓度的增加,脯氨酸的含量和脯氨酸合成关键酶基因的表达显著地增强;(3)Na+和脯氨酸是植物有效的渗透调节剂,可使处于低水势的植物细胞仍能从细胞外高浓度的盐溶液中吸收水分;(4)在0和700 mmol·L-1Na Cl处理下,盐爪爪肉质化叶中丙二醛的含量较其它处理高,这表明植物在这两个处理下可能受到了氧化胁迫;(5)从盐胁迫3个月的生长表型来看,低盐环境中生长的盐爪爪植株的生物量更多,肉质化的叶嫩且绿。综上所述,结合对野外生境的调查和实验室长期的盐胁迫表型结果表明盐爪爪的生长是需盐的,相对低的盐浓度环境对盐爪爪的生长是顺境,而无盐或高浓度盐环境对于盐爪爪的生长来说都是逆境。该研究结果为全面深入研究盐爪爪的耐盐特性,以及更好地利用盐爪爪的生物和基因资源改良土壤和提高作物和林木的耐盐性奠定基础。     

盐生植物海马齿耐盐的生理特性

以盐生植物海马齿为研究材料,分别用淡水、1/4海水、1/2海水、全海水浇灌15 d和30 d,研究盐生植物耐盐的生理特性和机理。海马齿植物在低于1/2的海水浇灌时,植物生长旺盛,主要表现为叶片增大和变厚,地上部分生物量增加;而全海水抑制了植物的生长。在盐胁迫下,海马齿植物中Na+的含量叶中最高,茎中含量次之,根中含量最低。长时间盐胁迫时,海马齿植物根、茎、叶中的相对含水量与淡水浇灌相比,变化不大,叶中略有增加;而脯氨酸含量显著增加,且可溶性糖的含量也比淡水浇灌的高。由此推测:海马齿植物主要以有机小分子作为渗透调节物质来维持细胞渗透压,在其耐盐中起着重要的作用。土壤中Na+的毒害,并没有减少土壤中可被植物利用的可交换K+,反而使其增加,说明海马齿植物根部对Na+的吸收能力和Na+/K+交换能力非常强。海马齿植物耐盐性强,还表现为能阻止盐胁迫对植物细胞原生质膜的氧化损伤,不破坏植物叶片内叶绿素的合成,能基本维持植物茎、叶中K+和根、茎中Mg2+的相对稳定。     

盐胁迫诱导的植物细胞凋亡--植物抗盐的可能生理机制

近来的研究表明,一定条件的盐胁迫可导致植物细胞程序性死亡。本文利用DNALaddering、石蜡切片原位检测以及染色体涂片原位检测,从组织、细胞以及DNA等多个方面对盐胁迫下的玉米、水稻和烟草根尖细胞死亡作了研究,形态特别是生化方面的证据表明盐胁迫诱导的植物细胞凋亡可能在植物界具有一定的普遍性。但各个物种之间有一定差异。本实验结果对盐胁迫下的植物生理机制提供了新的研究思路。同时,我们还对基于染色体制片和石蜡切片的原位检测方法进行了比较和讨论。我们认为,基于染色体制片的原位标记技术适合于定性和定量检测单个细胞的凋亡,具有一些石蜡切片所不可及的优点。     

Dinitro-o-cresol induces apoptosis-like cell death but not alternative oxidase expression in soybean cells

In plants, programmed cell death is thought to be activated during differentiation and in response to biotic and abiotic stresses. Although its mechanisms are far less clear, several morphological and biochemical features have been described in different experimental systems, including DNA laddering and cytosolic protease activation. Moreover, plant mitochondria have an alternative terminal oxidase (AOX), which is thought to be involved in protection against increased reactive oxygen species production, perhaps representing a mechanism to prevent programmed cell death. In this study, we analysed cell death induced by the herbicide dinitro-o-cresol (DNOC) in soybean (Glycine max) suspension cell cultures and evaluated biochemical and molecular events associated with programmed cell death. AOX capacity and expression were also determined. DNOC-treated cells showed fragmented nuclear DNA as assessed by an in situ assay that detects 3'-OH ends. In addition, specific colorimetric assays and immunoblot analysis revealed activation of caspase-3-like proteins and release of cytochrome c from mitochondria, respectively, confirming the apoptotic-like phenotype. Surprisingly, AOX capacity and protein levels decreased in DNOC-treated cells, suggesting no association between cell death and AOX under these experimental conditions. In conclusion, the results show that DNOC induces programmed cell death in soybean cells, suggesting that plants and animals might share similar pathways. Further, the role of AOX in cell death has not been confirmed, and may depend on the nature and intensity of stress conditions.     

Microwave irradiation of paraffin-embedded tissue sensitizes the TUNEL method for in situ detection of apoptotic cells

Apoptosis is a morphologically distinct form of programmed cell death that plays an important role in the growth regulation of a variety of tissues and also in the elimination of self-reacting immunocompetent cells. Several techniques for the qualitative and quantitative detection of this process have been established; recently, an in situ nick end-labelling technique based on the detection of DNA fragmentation, which is a molecular characteristic of apoptotic cell death, was described. Applying this method to paraffin sections of human tissues, sensitivity was observed to be inconsistently low with regard to the expected number of apoptotic cells. In the present study we show that irradiation of the tissue sections in 10 mM citrate buffer, pH 6.0, by microwaves at 750 W considerably enhances the sensitivity of this nick end-labelling technique.     

On the use of an appropriate TdT-mediated dUTP–biotin nick end labeling assay to identify apoptotic cells

Apoptosis is an essential cellular mechanism involved in many processes such as embryogenesis, metamorphosis, and tissue homeostasis. DNA fragmentation is one of the key markers of this form of cell death. DNA fragmentation is executed by endogenous endonucleases such as caspase-activated DNase (CAD) in caspase-dependent apoptosis. The TUNEL (TdT-mediated dUTP–biotin nick end labeling) technique is the most widely used method to identify apoptotic cells in a tissue or culture and to assess drug toxicity. It is based on the detection of 3′-OH termini that are labeled with dUTP by the terminal deoxynucleotidyl transferase. Although the test is very reliable and sensitive in caspase-dependent apoptosis, it is completely useless when cell death is mediated by pathways involving DNA degradation that generates 3′-P ends as in the LEI/L-DNase II pathway. Here, we propose a modification in the TUNEL protocol consisting of a dephosphorylation step prior to the TUNEL labeling. This allows the detection of both types of DNA breaks induced during apoptosis caspase-dependent and independent pathways, avoiding underestimating the cell death induced by the treatment of interest.     

Mycotoxins reveal connections between plants and animals in apoptosis and ceramide signaling

Plants undergo programmed cell death during development and disease in contexts that are functionally analogous to apoptosis in animals. Recent studies involving plant cell death induced by mycotoxins, pathogens and lethal mutations along with the cell-autonomous death during development now point to several conserved connections to apoptosis in animals. Morphological markers indicative of apoptosis recently reported in plants include TUNEL positive cells, DNA ladders, Ca2+-activated nucleosomal DNA cleavage, and formation of apoptotic-like bodies that occur in some but not all situations involving ordered cell death. In parallel studies with animal and plant cells treated with sphinganine analog mycotoxins our results indicate that the induction and inhibition of death may be mediated by ceramide-linked signaling systems. The presence and significance of ceramide-linked second messenger systems is well documented in animals but is virtually unknown in plants. Further research will discern the manner in which the important function of programmed cell death is conserved as well as diverged between the two kingdoms.     

MEK/ERK inhibitor U0126 enhanced salt stress-induced programmed cell death in <Emphasis Type="Italic">Thellungiella halophila</Emphasis> suspension-cultured cells

Programmed cell death (PCD) is an active cellular suicide that occurs both in animals and plants throughout development and in response to abiotic or biotic stress. In contrast to plant hypersensitive response-like cell death, little is known about the molecular machinery that regulates the halophyte plant PCD under high salinity stress. Since mitogen-activated protein kinases (MAPKs) are involved in plant response/tolerance to salt stress, and plant MAPK genes belong to the extracellular signal-regulated kinase (ERK) subfamily, we have investigated the role of ERK-like enzymes in high salinity stress-induced cell death in Thellungiella halophila. The data showed that ERK-like enzymes were early (10 min) and transiently activated under 300 mM NaCl stress. Pretreatment with 10 μM U0126, a special MEK/ERK inhibitor, resulted in a small but statistically significant increase of the percentage of terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (TUNEL)-positive nuclei in contrast to salt alone. The effects of U0126 on H₂O₂ production and cytochrome c (cyt c) release were also investigated. We found that the pretreatment with U0126 accelerated H₂O₂ production as well as cyt c release, and eventually enhanced cell death. The results suggest that ERK-like enzymes in Thellungiella halophila may act as a positive regulator of salt tolerance, as illustrated by pretreatment with U0126 which enhanced cell death under high salinity stress.     

Identification of programmed cell death in situ in individual plant cells in vivo using a chromosome preparation technique

A simple procedure, which combines a chromosome preparation technique with an in situ labelling technique modified from fluorescence in situ hybridization (FISH), has been developed for in situ detection of plant programmed cell death (PCD) at the single-cell level. After exposure of chromosomes and nuclei on slides by enzymolysis, Klenow or TdT was used to incorporate Bio-dUTP or fluorescein-dUTP at sites of DNA breaks. After Klenow-mediated labelling, the signals were amplified by a cascade of antigen-antibody reaction according to the detection system of FISH. This method enables in situ detection of plant PCD in vivo morphologically and biochemically at the chromosome, nuclear and DNA levels without cell culture and histological sectioning. This technique permits labelling of DNA breaks with high sensitivity due to increased chromosome and nucleus exposure to the labelling solutions, as well as due to the immunological amplification of the signals. Moreover, the changes in the cells were easier to be observed because the spatial obstacle of the cell wall and its autofluorescence were eliminated. It is potentially useful for in situ detection of PCD in plant root meristematic cells triggered by various environmental abiotic factors. It is proposed that the root tip is a versatile in vivo system for studying PCD induced by environmental abiotic factors.     

Protective effects of resveratrol on hydrogen peroxide-induced apoptosis in rat pheochromocytoma (PC12) cells

Oxidative stress has been considered as a major cause of cellular injuries in a variety of clinical abnormalities. One of the plausible ways to prevent the reactive oxygen species (ROS)-mediated cellular injury is dietary or pharmaceutical augmentation of endogenous antioxidant defense capacity. Resveratrol (3,5,4'-trihydroxy-trans-stilbene), one of the major antioxidative constituents found in the skin of grapes, has been considered to be responsible in part for the protective effects of red wine consumption against coronary heart disease ('French Pardox'). In this study, we have investigated the effects of resveratrol on hydrogen peroxide-induced oxidative stress and apoptotic death in cultured rat pheochromocytoma (PC12) cells. PC12 cells treated with hydrogen peroxide underwent apoptotic death as determined by characteristic morphological features, internucleosomal DNA fragmentation and positive in situ end-labeling by terminal transferase (TUNEL staining). Resveratrol pretreatment attenuated hydrogen peroxide-induced cytotoxicity, DNA fragmentation, and intracellular accumulation of ROS. Hydrogen peroxide transiently induced activation of NF-kappaB in PC12 cells, which was mitigated by resveratrol pretreatment. These results suggest that resveratrol has the potential to prevent oxidative stress-induced cell death.     

Karyotyping of Brassica oleracea L. based on rDNA and Cot-1 DNA fluorescence in situ hybridization

To explore an effective and reliable karyotyping method in Brassica crop plants,Cot-1 DNA was isolated from Brassica oleracea genome,labeled as probe with Biotin-Nick Translation Mix kit,in situ hybridized to mitotic spreads,and where specific fluorescent bands showed on each chromosome pair.25S and 5S rDNA were labeled as probes with DIG-Nick Translation Mix kit and Biotin-Nick Translation Mix kit,respectively,in situ hybridized to mitotic preparations,where 25S rDNA could be detected on two chromosome pairs and 5S rDNA on only one.Cot-1 DNA contains rDNA and chromosome sites identity between Cot-1 DNA and 25S rDNA was determined by dual-colour fluorescence in situ hybridization.All these showed that the karyotyping technique based on a combination of rDNA and Cot-1 DNA chromosome landmarks is superior to all but one.A more exact karyotype ofB.oleracea has been analyzed based on a combination of rDNA sites,Cot-1 DNA fluorescent bands,chromosome lengths and arm ratios.