Dissecting Multiple Arabidopsis CC-NBS-LRR Proteins Structure and Localization

NBS-LRR (nucleotide binding sites and leucine rich repeat) protein plays a crucial role as sentries and as defense activators in plants. The structure and function of NBS-LRR proteins are closely related. Previous articles have announced that the activated ZAR1 (HopZ-Activated Resistance 1) forms a pentamer in the plasma membrane, which is a calcium permeable channel that can trigger plant immune signaling and cell death. However, the structure of galore NBS-LRRs in Arabidopsis is not yet clear. The functional sites of distinct NBS-LRR in cells may vary. In addition, identifying pathogens and activating defense regions may occur in different subcellular compartments. Therefore, dissecting the specific structure and positioning of NBS-LRRs is an indispensable step in understanding their functions. In this article, we exploit AlphaFold to predict the structure of some designed NBS-LRRs, and utilize Agroinfiltration transient expression system, combined with biochemical fractionation, to dissect the localization of these NBS-LRR receptors from Arabidopsis. Structural data indicates that the identified NBS-LRRs share analogous conformation. Membrane fractionation assay demonstrates these NBS-LRRs are mainly associated with the membrane. These data show that the Ca2+-permeable channel activity may be evolutionarily conserved in NBS-LRR of Arabidopsis, and this study provides some reference clues for analyzing the structure and localization patterns of other plant immune receptors.

The NAC Transcription Factor ANAC089 Modulates Seed Vigor through the ABI5-VTC2 Module in Arabidopsis thaliana

Seed viability is an essential feature for genetic resource conservation as well as sustainable crop production.Long-term storage induces seed viability deterioration or seed aging,accompanied by the accumulation of toxic reactive oxygen species(ROS)to suppress seed germination.Controlled deterioration treatment(CDT)is a gen-eral approach for mimicking seed aging.The transcription factor ANAC089 was previously reported to modulate seed primary germination.In this study,we evaluated the ability of ANAC089 to control seed viability during aging.Compared with that in the wild-type line,the mutation of ANAC089 significantly increased H_(2)O_(2),thereby reducing seed viability after CDT,while the overexpression of ANAC089 reduced H_(2)O_(2) and improved seed long-evity,indicating a critical role for ANAC089 in maintaining seed viability through H_(2)O_(2) signaling.A series of stu-dies have shown that ANAC089 targets and negatively regulates the level of ABI5,an important transmitter of abscisic acid(ABA)signals,to affect seed viability after CDT.Furthermore,ABI5 negatively regulated the expres-sion of VTC2,which is involved in the biosynthesis of the antioxidant ascorbic acid and H_(2)O_(2) scavenging.As a result,ANAC089 attenuates the generation of H_(2)O_(2),thereby enhancing seed viability through the ABI5-VTC2 module during the seed aging process.Taken together,our results reveal a novel mechanism by which ANAC089 enhances seed viability by coordinating ABI5 and VTC2 expression,ultimately preventing the overac-cumulation of H_(2)O_(2),which would have led to reduced seed viability.

Molecular and genetic regulations of fleshy fruit shape and lessons from Arabidopsis and rice

Fleshy fruit shape is an important external quality trait influencing the usage of fruits and consumer preference.Thus,modification of fruit shape has become one of the major objectives for crop improvement.However,the underlying mechanisms of fruit shape regulation are poorly understood.In this review we summarize recent progress in the genetic basis of fleshy fruit shape regulation using tomato,cucumber,and peach as examples.Comparative analyses suggest that the OFP-TRM(OVATE Family Protein-TONNEAU1 Recruiting Motif)and IQD(IQ67 domain)pathways are probably conserved in regulating fruit shape by primarily modulating cell division patterns across fleshy fruit species.Interestingly,cucumber homologs of FRUITFULL(FUL1),CRABS CLAW(CRC)and 1-aminocyclopropane-1-carboxylate synthase 2(ACS2)were found to regulate fruit elongation.We also outline the recent progress in fruit shape regulation mediated by OFP-TRM and IQD pathways in Arabidopsis and rice,and propose that the OFP-TRM pathway and IQD pathway coordinate regulate fruit shape through integration of phytohormones,including brassinosteroids,gibberellic acids,and auxin,and microtubule organization.In addition,functional redundancy and divergence of the members of each of the OFP,TRM,and IQD families are also shown.This review provides a general overview of current knowledge in fruit shape regulation and discusses the possible mechanisms that need to be addressed in future studies.

Glucosinolates and Their Hydrolysis Products in Arabidopsis thaliana Influence Performance and Feeding Choice of Pieris rapae and Spodoptera exigua

Glucosinolates and their hydrolysis products, found in plants of the order Brassicales, are well-known for their defensive properties against insect herbivores. Arabidopsis thaliana (Col-0) genetic lines with mutations that modify the type of glucosinolates (i.e. myb28myb29 and cyp79B2cyp79B3 are deficient in the production of aliphatic and indolyl glucosinolates, respectively) make it possible to test for the specific effects of these secondary chemicals on insect herbivores. The Pad3 mutant (deficient in camalexin), which has a role in resistance to pathogens, was also tested. Likewise, the effects of different glucosinolate hydrolysis products can be evaluated using genetically modified (GM) lines of the wild type Col-0 ecotype, which naturally produces isothiocyanates. These GM lines include the nitrile-producing 35S: ESP and the double knockout tgg1tgg2, which virtually lacks hydrolysis products. In both no-choice and choice experiments, the crucifer specialist Pieris rapae was virtually unaffected by differences in the type of glucosinolates or hydrolysis products. In contrast, the generalist insect Spodoptera exigua had statistically significant increases in pupae/adult weight and faster developmental times when reared on mutants deficient in the production of aliphatic and indolyl glucosinolates and their hydrolysis products. There were no differences in the performance of either insect species when reared on wild type Col-0 or Pad3. Results from feeding choice trials showed that Pieris rapae had no statistically significant preference for any of the genetic lines. In contrast, Spodoptera exigua had a significant feeding preference for the double mutant tgg1tgg2. This study provides evidence that variation in the type of glucosinolates and their hydrolysis products can influence insect performance and feeding choices, and that responses are species-specific.

Overexpression of Wheat TaELF3-1BL Delays Flowering in Arabidopsis

EARLY FLOWERING 3(ELF3),a light zeitnehmer(time-taker)gene,regulates circadian rhythm and photoperiodic flowering in Arabidopsis,rice,and barley.The three orthologs of ELF3(TaELF3-1AL,TaELF3-1BL,and TaELF3-1DL)have been identified in wheat too,and one gene,TaELF3-1DL,has been associated with heading date.However,the basic characteristics of these three genes and the roles of the other two genes,TaELF3-1BL and,TaELF3-1AL,remain unknown.Therefore,the present study obtained the coding sequences of the three orthologs(TaELF3-1AL,TaELF3-1BL,and TaELF3-1DL)of ELF3 from bread wheat and characterized them and investigated the role of TaELF3-1BL in Arabidopsis.Protein sequence comparison revealed similarities among the three TaELF3 genes of wheat;however,they were different from the Arabidopsis ELF3.Real-time quantitative PCR revealed TaELF3 expression in all wheat tissues tested,with the highest expression in young spikes;the three genes showed rhythmic expression patterns also.Furthermore,the overexpression of the TaELF3-1BL gene in Arabidopsis delayed flowering,indicating their importance in flowering.Subsequent overexpression of TaELF3-1BL in the Arabidopsis ELF3 nonfunctional mutant(elf3 mutant)eliminated its early flowering phenotype,and slightly delayed flowering.The wild-type Arabidopsis overexpressing TaELF3-1BL demonstrated reduced expression levels of flowering-related genes,such as CONSTANS(AtCO),FLOWERING LOCUS T(AtFT),and GIGANTEA(AtGI).Thus,the study characterized the three TaELF3 genes and associated TaELF3-1BL with flowering in Arabidopsis,suggesting a role in regulating flowering in wheat too.These findings provide a basis for further research on TaELF3 functions in wheat.

Biochemical and Physiological Responses of Arabidopsis thaliana Leaves to Moderate Mechanical Stimulation

Mechanical stimulation of plants can be caused by various abiotic and biotic environmental factors.Apart from the negative consequences,it can also cause positive changes,such as acclimatization of plants to stress conditions.Therefore,it is necessary to study the physiological and biochemical mechanisms underlying the response of plants to mechanical stimulation.Our aim was to evaluate the response of model plant Arabidopsis thaliana to a moderate force of 5 N(newton)for 20 s,which could be compared with the pressure caused by animal movement and weather conditions such as heavy rain.Mechanically stimulated leaves were sampled 1 h after exposure and after a recovery period of 20 h.To study a possible systemic response,unstimulated leaves of treated plants were collected 20 h after exposure alongside the stimulated leaves from the same plants.The effect of stimulation was assessed by measuring oxidative stress parameters,antioxidant enzymes activity,total phenolics,and photosynthetic performance.Stimulated leaves showed increased lipid peroxidation 1 h after treatment and increased superoxide dismutase activity and phenolic oxidation rate after a 20-h recovery period.Considering photosynthetic performance after the 20-h recovery period,the effective quantum yield of the photosystem II was lower in the stimulated leaves,whereas photochemical quenching was lower in the unstimulated leaves of the treated plants.Nonphotochemical quenching was lower in the stimulated leaves 1 h after treatment.Our study suggested that plants sensed moderate force,but it did not induce pronounced change in metabolism or photosynthetic performance.Principal component analysis distinguished three groups–leaves of untreated plants,leaves analysed 1 h after stimulation,while stimulated and unstimulated leaves of treated plants analysed 20 h after treatment formed together the third group.Observed grouping of stimulated and unstimulated leaves of treated plants could indicate signal transduction from the stimulated to distant leaves,that is,a systemic response to a local application of mechanical stimuli.

不同栽培密度下拟南芥(Arabidopsis thaliana)的形态分析

为初步阐明双子叶植物响应栽培密度变化的规律,本研究以模式植物拟南芥(Arabidopsis thaliana)为材料,设置不同栽培密度(1×1、2×2、3×3、4×4、5×5、6×6、7×7)进行盆栽试验,分析不同栽培密度下拟南芥不同生长时期的形态指标变化规律。结果表明:(1)在同一栽培密度处理下,随着拟南芥的生长发育,其株高、茎长和根长逐渐增加,莲座叶直径逐渐增加直至稳定,叶面积先增大后减小,顶部分枝数和基部分枝数逐渐增加或逐渐增加直至稳定。在同一生长时期,拟南芥个体大小随着栽培密度增加而减小,且高栽培密度下抽薹和开花时间提前。(2)不同栽培密度对拟南芥株高、茎长、根长、莲座叶直径、顶部分枝、基部分枝和叶面积均存在显著影响。随着栽培密度的增加,拟南芥株高、茎长总体呈降低趋势,根长呈逐渐增加趋势,莲座叶直径、顶部分枝数和基部分枝数呈逐渐减小趋势,叶面积表现为先增大后减小或逐渐减小趋势。本试验从形态上初步阐明了拟南芥响应栽培密度变化的规律,栽培密度对不同生长时期拟南芥各形态指标均产生了显著影响,该结果可为通过合理密植调控双子叶植物形态提供理论依据。

新疆短命植物小拟南芥(Arabidopsis pumila)种子萌发特性及其生态适应性

通过显微结构及不同处理条件下种子萌发率的观察,对早春短命植物小拟南芥(Arabidopsis pumila)种子萌发特性及影响因素进行了研究,并对其生态适应性进行了讨论。结果表明:(1)温度和光照变化对自然生境和温室收获种子的萌发率影响均不显著,说明此种群在前期萌发阶段对光、温不敏感;(2)自然生境中采收的小拟南芥种子萌发率显著低于温室收获种子,说明环境条件的变化对短命植物种子的发育具有重要作用,可显著改变种子的萌发行为;(3)赤霉素使自然生境收获种子胚活性增强从而对萌发有较大促进作用,可使萌发率增加50%以上;(4)对种皮进行各种机械损伤处理使得种皮松弛或透气,可以显著提高自然生境种子的萌发率(超过70%);(5)盐和干旱胁迫对种子萌发均具有明显的抑制作用,但复水后部分被抑制种子可重新萌发,显示盐和干旱胁迫可导致种子产生浅度休眠。结合小拟南芥自然生存环境及本研究的结果,显示其种子萌发特性与生境具有高度适应性。

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.

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.

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.

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.

不同光照条件下新疆小拟南芥(Arabidopsis Pumila)的表型可塑性研究

田间观测同一区域不同光照条件下小拟南芥可塑性变化,研究表明:1)阴阳两地小拟南芥在株高和茎生叶数、叶重、花序重、茎重、莲座叶重等特征均达到显著差异,但是两地适合度差异不显著,显示出很强的可塑性;2)阴阳两地小拟南芥在叶重与茎生叶数、花序重与花序长、茎重与株高、地下重与主根长、角果重与角果数均表现为极显著相关,表现出很强的特征整合能力。根据现代植物可塑性研究成果,对新疆境内小拟南芥光适应性特点及进化趋势进行了分析。

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

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.

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.

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.