天山北麓中段拟南芥(Arabidopsis thaliana)与相邻物种的分布格局及相互关系

天山山脉是世界拟南芥(Arabidopsis thaliana)及其近缘种的分布中心之一,资源优势明显。在北天山中段浅山地带选择拟南芥分布的典型样地50m×50m,分析了样地物种的结构、组成和土壤理化性质,用Ripley’sK(d)函数分析了拟南芥与相邻物种的空间特征和相互关系。发现样地由7科23个物种组成,以新疆绢蒿(Seriphidium kaschgaricum)为建群种,短命植物物种占近70%。拟南芥仅分布于北坡,在3m内聚集强度高于所有分析物种,在5m范围内与新疆绢蒿中株呈显著正关联,与十字花科的涩芥(Malcolmia africana)、藜科的散枝猪毛菜(Salsola brachiata)、木碱蓬(Suaeda dendroides)、角果藜(Ceratocarpus arenarius)呈一定尺度显著负关联。分析认为拟南芥空间分布依赖于新疆绢蒿大株、中株生长塑造的遮阴、保湿和丰富土壤有机质,生态位与藜科物种差异极大,生境特异性高于同属近缘种小鼠耳芥(Arabidopsis pumila),以及涩芥(M.africana)、庭芥(Alyssumdesertorum)、四齿芥(Tetracme quadricornis)、丝叶芥(Leptaleum filifolium)、狭果鹤虱(Lappula semiglabra)等短命植物。在干旱胁迫下,拟南芥环境选择强度大于种内作用,密度依赖的种子扩散表现不明显。扩散对策是通过大量生产种子,依靠果实不易开裂控制种子短距离扩散,充分利用原适宜生境来维持种群繁衍。

生态因子及其交互作用对拟南芥(Arabidopsis thaliana)表型可塑性的影响

以拟南芥(Arabidopsis thaliana)两种基因型(ws-0和col-0)材料,采用复因子混合水平正交试验设计开展盆栽实验,研究了土壤盐分、土壤水分、光照强度、去叶处理等生态因子及其交互作用对受试植株18个表型特征的影响。结果表明生态因子对植物表型可塑性的影响是有针对性的:土壤水分主要影响植物体构件数目;土壤盐分主要影响生物量、角果数及种籽总数等直接反映植株适合度的表型特征;光照条件则主要影响植物的物候表型特征。植物体表型可塑性的方向随水分梯度的变化而发生改变。生态因子交互作用对植物表型可塑性的影响效果不是各因子独立作用的简单加和:对某个表型特征都有显著影响的两个生态因子其交互作用对该特征可能没有影响;反之,受两个生态因子交互作用影响显著的表型特征也可能不受它们的独立影响。在对生态因子交互作用作出响应时,col-0的9个特征表现出可塑性,而ws-0仅有4个表型是可塑的;同一基因型内彼此相关的表型特征在可塑性上也具一致性。抽苔时莲座叶数与角果平均籽粒数不受任何生态因子及其交互作用的影响,这两个表型作为数量特征而未表现出可塑性。

Physiological Analysis of Two Arabidopsis thaliana Mutants in Response to CO2

[Objective] The 15urpose was to seek for the different phenotypes between wild type and Arabidopsis Mutants in response to CO2. [Method] The epidermis bioassays and seed germination test were carried out to analyze the physiological characteristics of two Arabidopsis mu- tants and their wild type. [Result] There existed distinct differences in stomata apertures, water loss and leaf temperature compared with wild type except for stomata density. In addition, seed germination test on the medium indicated that cdfl was insensitive to ABA, mannitol and NaCI, but cdsl performed contrary to cdil. [ Conclusion] There are some different physiological characteristics between wild type and mutants.

拟南芥(Arabidopsis thaliana)氮、碳离子注入诱变效应分析

采用N +、C +离子注入拟南芥 (Arabidopsisthaliana)种子 ,统计了种子的发芽指数 (发芽率和发芽势 ) ;用改良的RAPD技术对N+离子注入种子植株的DNA进行11个引物的随机片段多态性扩增。结果表明 ,合适剂量的N+、C+离子注入可使种子发芽率提高 ,两种离子注入种子的发芽率峰值 (分别为92.3 %和74.4 % )都在5×1014ions/cm2;分析N +离子注入材料发现 ,在1×1013 -1×1016ions/cm2剂量范围内 ,基因组DNA的变异率与发芽指数的变化趋势基本一致 ,变异率峰值 (9.0 % )在1×1015 ions/cm2。结果提示 ,分析低能N+离子诱变效应的最佳注入剂量在1×1014-5×1015 ions/cm2。对N+、C+离子注入的比较发现 ,一定范围内同等剂量C+离子注入的诱变率高于N

A Rapid DNA Extraction Method for PCR Detection of Arabidopsis thaliana

[Objective] The aim was to introduce a rapid DNA extraction method for PCR detection of Arabidopsis thaliana.[Method] Through the improvement of conventional DNA extraction method,a rapid Arabidopsis thaliana DNA extraction method was obtained.With randomly selected Arabidopsis thaliana transgenic strains and mutants as samples,the method was verified.[Result] After electrophoresis,UV absorption detection,it was found that DNA samples are complete and less pollution,and the result of PCR amplification objective fragment was good which proved DNA is suitable as a template for PCR reaction.After PCR detection,positive plants gene amplified bands were clear,without false-positive,and the test results were satisfactory.[Conclusion] The method is suitable for rapid extraction of Arabidopsis thaliana DNA and PCR detection.

Effects of Drought Stress Simulated by PEG on Glucosinolates Content in Arabidopsis thaliana

[Objective] The paper aimed to study effects of drought stress simulated by PEG on glucosinolates content in Arabidopsis thaliana.[Method] Drought stress was simulated by PEG-6000,ecological seeds of Arabidopsis thaliana were cultivated by the control group and drought treatment group respectively,Physical signs of Arabidopsis thaliana and contents of glucosinolates were determined after 0,4,5,6,7 d treatment.[Result] The results showed that leaf water content of rosette leaves was obviously decreased,leaf relative conductivity （characterized by membrane permeability） and the concentration of MDA increased,the extent of damage increased with the increased time.Content of total glucosinolate,aliphatic glucosinolate and indole glucosinolate increased got their maximum after 5 days treatment,and rapidly decreased after 6 and 7 days of treatment,even much lower than the control group.Each kind of glucosinolate changed with difference from each other.4MSOB which made the most proportion of the total glucosinolate changed consistently with the total glucosinolate and difference significant.As a whole,aliphatic glucosinolates were more sensitive to drought than indole glucosinolate.The proportion of some kind glucosinolate,like 4MSOB varied had correlation with the content change.[Conclusion] Drought stress had an effects on the contents of total glucosinolate,aliphatic glucosinolate and indole glucosinolate,which made the glucosinolate participated in defense response of plant to the outside of drought stress,but long-term drought stress in leaves was not conducive to the accumulation of glucosinolates.

NaCl对拟南芥(Arabidopsis thaliana)生长与渗透调节的影响

对与拟南芥耐盐性相关的某些生理指标进行了研究。结果发现,随着盐浓度的增加,拟南芥的质膜透性增加,MDA含量、脯氨酸含量、可溶性糖含量、有机酸含量、游离氨基酸含量、根冠比、钠离子含量均增加,而渗透势、钾离子含量、干鲜重、含水量则下降。这表明,随着盐胁迫的加重,一方面拟南芥受到的伤害加重,另一方面植株通过合成脯氨酸、可溶性糖等渗透调节物质,增大根冠比以提高吸水能力等抗逆机制,以应对胁迫,提高耐盐性。

Ultrastructural Studies on the Sieve Elements in Root Protophleom of Arabidopsis thaliana

The ultrastructures of the root protophleom sieve element at different developmental stages of Arabidopsis thaliana L. were investigated using the technique of high pressure freezing and freeze substitution fixing specimen. The results show that in the development of the sieve elements, the nuclei undergo typical characteristics of the programmed cell death (PCD): the nuclear envelopes form emboli, the chromatin condenses and aggregates towards the nuclear envelope, which degrades and fully disappears later. Before the nucleus degradation, neither the nuclear envelope undulation, nucleus lobe nor marked dilation (or bleb) of perinuclear space could be observed. In the cytoplasm of the mature sieve element, there are starch-like granules separately sheathed with a layer of membrane and usually with mitochondria around. These gnanules seem to provide substrates to mitochondria in their function. Small vacuoles originate from endoplasmic reticulum (ER), and no bigger vacuole was found.

Different Ecological FLC Gene in the Arabidopsis thaliana Sequence Analysis

[Objective]Analysis of FLC sequence that Vernalization-related genes in Arabidopsis.[Method]Advance through natural populations of Arabidopsis QTL analysis of vernalization response was found on chromosome 5 of Arabidopsis thaliana have a flowering-related QTL,this test is to use sequence analysis to determine whether it is with the FLC gene homology.[Result]Arabidopsis thaliana,Italy and Sweden in the 27th,No.461,p.501,p.638,p.738,No.884 different base.While these bases are different,but the codon encoding the first nine amino acids out of the first 167 amino acids,the first 246 amino acids,due to codon degeneracy,encode proteins that are the same.[Conclusion] Arabidopsis has a rich genetic diversity,the FLC gene is highly conserved sequence length,base variable sites rich degenerate codons encoding the same amino acids they are not affect the growth of Arabidopsis thaliana.This indicates that the Arabidopsis genome sequence will be the environment.

Cytological Studies on the Development of Sieve Element and Floral Nectary Tissue in Arabidopsis thaliana

Ultrastructural changes in the sieve element and dense cells of nectariferous tissue during the development of floral nectary in Arabidopsis thaliana L. were investigated with transmission electron microscopy. Samples were prepared with high pressure freezing and freeze substitution techniques. The ultrastructure of dense cell was similar to that of sieve element at its early developmental stage. With the concurrent agglutination of chromatin in the nucleus, the abnormal location of organelles and the high density of cytoplasm, the ultrastructural characteristics in die dense cells of the nectariferous tissue and in the sieve element are matched with those of the programmed cell death in animal and plant reported in recent years. The disorganization of nucleus and most organelles in the differentiation of sieve elements and dense cells is closely associated with the transportation and modification of pre-nectar and the transference of nectar. This suggests that the cytological changes in sieve element and nectariferous tissue are closely associated with the nectary functional activities.

Effects of Salt Stress on Epidermal Cell Expansion in Leaves of Arabidopsis thaliana

[Objective] The aim of this study was to investigate the effects of salt stress on cell expansion in Arabidopsis thaliana rosette leaves.[Method] Arabidopsis seedlings were treated by sodium chloride at the concentration of 0,100 or 150 mmol/L. At the 7th and 14th d of treatment,with nail enamel printing mark method and computer software,the leaf blades area and abaxial epidermal pavement cells area was measured and compared using statistical analysis in Excel. [Result] The growth of Arabidopsis rosette leaves was inhibited under salt stress. Leaves treated for 7 or 14 d expanded less compared with controls. The salt-mediated decrease in leaf expansion is associated with a decrease in abaxial pavement cell expansion. [Conclusion] The decreased leaf and epidermal cell expansion under salt stress is the most important characteristic of plant physiological response to salt stress.

Study on the Callus Induction of Wild Arabidopsis thaliana Leaves

[Objective] The callus induction of wild Arabidopsis thaliana leaves was studied.[Method] As explants,leaf segments of wild A.thaliana were inoculated in MS medium including 6-BA and NAA with different concentration,and studying the callus induction and regenerated plant of A.thaliana.[Result]6-BA was necessary for callus induction,but higher concentration resulted into vitrification easily;single use of NAA was beneficial to rooting,and bud differentiation was easy in the medium with NAA and 6-BA;the optimum medium of callus induction was MS ＋0.5 mg/L 6-BA ＋0.10 mg/L NAA,with the callus induction frequency of 100%.[Conclusion]Our study could lay a foundation for the genetic transformation and cell culture of A.thaliana.

Regulation of Heat Shock Factor AtHsfA1a on Ascorbate Peroxidase under Heat Stress in Arabidopsis thaliana

[Objective] This study was conducted to investigate the regulation of heat shock factor AtHsfA1a on ascorbate peroxidase under heat stress in Arabidopsis thaliana. [Method] After heat stress treatment on transgenetic A. thaliana with silenced endogenetic AtHsfA1a gene and wild A. thaliana plants as materials, the change in activity of APX enzyme was analyzed by spectrophotometry, the expression level of APX gene was investigated by real-time fluorescent quantitative PCR, and the binding condition of AtHsfAla with the promoter region of APX gene was analyzed by chromatin immunoprecipitation assay. [Result] The activity and mRNA level of APX in plants with silenced endogenetic AtHsfAla gene were higher than those in wild plants. Fragments of the promoter region of APX gene were not screened from the plants with silenced endogenetic AtHsfA1a gene, but found in wild plants. [Conclusion] This study provides a theoretical basis for the understanding of the important role of AtHsfAla in resistance to stress in plant, and is of great significance to the revealing of mechanism of resistance to stress in plant.

Cloning and Expression Analysis of the High Mobility B Group Genes in Arabidopsis thaliana

[Objective] The aim was to better research the function and action mode of High Mobility Group B （HMGB） proteins in higher plants. [Method] At2G33450,At5G23405 and At5G23420 genes of HMGB protein family in Arabidopsis thaliana were cloned by the use of RT-PCR method,and the expression of these three proteins in E.coli and Arabidopsis thaliana were detected by using SDS-PAGE,Northern blot and subcellular localization methods. [Result] The results showed that the molecular weights of the three proteins were 17.5,17.0 and 27.0 kD respectively,and the expression levels of the proteins in Arabidopsis thaliana were At5G23420At5G23405At2G33450. In addition,all the three proteins were located in nucleus. [Conclusion] The study will provide a basis for the further research on the biological function of HMGB proteins in higher plants.

Quercetin Promotes Auxin Transport in Arabidopsis thaliana

Study on the role of quereentin in polar auxin transportation. Arabidopsis was cultured on medium supplemented with quereetin to observe the growth of hypoeotyls, ^14C-IAA transport assays were conducted to measure the auxin transport activity. The results showed that Arabidopsis mutant auxl which had been deficient in auxin influx transportion obviously recovered the ability after eultured on the medium with quercetin. The polar auxin transport was promoted by the addition of quereetin. These results indicated that quereetin could promote polar auxin transport in vivo.

拟南芥(Arabidopsis thaliana)种群间及近远缘种表型整合的分化研究

表型整合是生物形态、生理和功能之间以及三者之内特征的相关性。表型整合的进化是生物进化研究的重要内容。拟南芥(Arabidopsis thaliana)是植物遗传、生理和进化生物学研究的模式植物,但目前采用现代比较方法,将拟南芥形态与生理功能特征相结合研究表型整合较少。本研究选取了石河子(新疆)、青河(新疆)、浙江、美国(哥伦比亚)等拟南芥四个种群,以及新疆境内小拟南芥(Arabidopsis pumila)、南芥(Arabis auriculata)和庭荠(Alys-sum desertorum)等拟南芥近远缘种作为实验材料,在同一环境下,研究形态、光合生理、生活史等特征,以及表型整合特征在种内和种间的分化特点。结果表明:石河子种群与另外三种种群之间的单个特征差异明显,各种群在两两特征之间相关格局分化较大。共同主成分分析(CPCA-Common Principal Component Analysis)发现石河子、青河、浙江三种中国种群的整合格局相同,而美国种群与其它三个拟南芥种群间的共同主成分相对较少;物种整合格局的分化与系统关系的远近有较大的关系,同时也与自然选择压紧密相关。

拟南芥(Arabidopsis thaliana)体细胞胚胎发生体系中的体细胞类减数分裂研究

在拟南芥生态型Ｌａｎｄｓｂｅｒｇ Ｅｒｅｃｔａ体细胞胚胎发生体系的胚性愈伤组织中观察到２种 类型的体细胞减数分裂现象。一种是体细胞染色体减数分组，其中，处于前期或中期的细胞染 色体分为２个或２个以上的组。其共同特点是，染色体直接分开，未观察到纺锤体，从染色体 的形态也看不出纺锤体的作用。染色体减数分组较多发生于多倍体细胞中。另一种类型是体 细胞减数分裂，这种类型类似于大小孢子发生过程的减数分裂，如第一次分裂前期也有染色 体的联会和配对。在脱分化培养基上的胚性愈伤组织中，单倍体细胞约占３％，四倍体细胞约 占４％。经体细胞类减数分裂产生的细胞都发生染色体重组。

Predicting Arabidopsis thaliana Gene Function by Transitiving Co-expression in Shortest-path

The present paper predicted the function of unknow genes by analyzing the co-expression data of Arabidopsis thaliana from biological pathway based on the shortest-path algorithm. This paper proposed that transitive co-expression among genes can be used as an important attribute to link genes of the same biological pathway. The genes from the same biological pathway with similar functions are strongly correlated in expression. Moreover,the function of unknown genes can be predicted by the known genes where they are strongly correlated in expression lying on the same shortest-path from the biological pathway. Analyzing the Arabidopsis thaliana from the biological pathway,this study showed that this method can reliably reveal function of the unknown Arabidopsis thaliana genes and the approach of predicting gene function by transitiving co-expression in shortest-path is feasible and effective.

Regulatory Function of Polyamine Oxidase-Generated Hydrogen Peroxide in Ethylene-Induced Stomatal Closure in Arabidopsis thaliana

Hydrogen peroxide （H2O2） is an important signaling molecule in ethylene-induced stomatal closure in Arabidopsis thaliana. Early studies on the sources of H2O2 mainly focused on NADPH oxidases and cell-wall peroxidases. Here, we report the involvement of polyamine oxidases （PAOs） in ethylene-induced H2O2 production in guard cells. In Arabidopsis epidermal peels, application of PAO inhibitors caused the failure of ethylene to induce H2O2 production and stomatal closure. Results of quantitative RT-PCR analysis and pharmacological experiments showed that AtPAO2 and AtPAO4 transcripts and activities of PAOs were both induced by ethylene. In transgenic Arabidopsis plants over-expressing AtPAO2 and AtPAO4, stomatal movement was more sensitive to ethylene treatment and H2O2 production was also significantly induced. The increased H2O2 production in the transgenic lines compared to the wild-type plants suggests that AtPAO2 and AtPAO4 probably are involved in ethylene-induced H2O2 production. Several factors which induce stomatal closure such as dehydration and high salinity all enhanced the expression of AtPAO2 and AtPAO4 to different degrees. Moreover, GFP- AtPAOs fusion protein localized in the nucleus, cytoplasm, and cell wall of the guard cells. Therefore, our results strongly indicated that PAO is a source of H2O2 generation in Arabidopsis guard cells and plays crucial roles in stomatal movement.

Abiotic Stresses and Phytohormones Regulate Expression of FAD2 Gene in Arabidopsis thaliana

Modification of unsaturated fatty acid （FA） levels has been found to accompany multiple abiotic stress acclimations in many plants. Delta 12 fatty acid desaturase （FAD2） plays a critical role in the synthesis of polyunsaturated FAs in plant cells by converting oleic acid （18：1） to linoleic acid （18：2）. To better understand the relationship between polyunsaturated FAs metabolism and stress adaptation, the expression of FAD2 gene and changes in the FA compositions under various abiotic stresses and phytohormone treatments in Arabidopsis thaliana was investigated in this study. A 1 423-bp promoter of the FAD2 gene was cloned and characterized from Arabidopsis. Several putative hormone- and stress- inducible cis-elements were identified in the cloned promoter, which include salt- and pathogen-inducible GT-1 motifs, low-temperature-responsive MYC element, dehydration-responsive MYB element, and GA signaling related WRKY71OS element. To investigate the fine regulation of FAD2 gene, a recombinant FAD2 promoter-GUS construct was introduced into Arabidopsis plants. Histochemical study showed that the promoter was ubiquitously active and responsive not only to exogenous phytohormones including ABA, 24-eBL, and SA but also to darkness, temperature, salt, and sucrose stresses in Arabidopsis seedlings. Consistent with the expression change, treatments with exogenous 24-eBL, ABA, SA, and NaCl resulted in reduction in polyunsaturated FAs in Arabidopsis seedlings. These findings suggest that the FAD2 gene with a wide variety of putative response elements in its promoter is responsive to multiple phytohormones and abiotic stresses and therefore may play an important role in stress responses of Arabidopsis during plant growth and seed development.