Early mannitol-triggered changes in the Arabidopsis leaf (phospho)proteome reveal growth regulators
Natalia NikonorovaLisa Van den BroeckShanshuo ZhuBrigitte van de CotteMarieke DuboisKris GevaertDirk InzéIve De Smet
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Abstract:
Leaf growth is a complex, quantitative trait, controlled by a plethora of regulatory mechanisms. Diverse environmental stimuli inhibit leaf growth to cope with the perceived stress. In plant research, mannitol is often used to impose osmotic stress and study the underlying growth-repressing mechanisms. In growing leaf tissue of plants briefly exposed to mannitol-induced stress, a highly interconnected gene regulatory network is induced. However, early signalling and associated protein phosphorylation events that probably precede part of these transcriptional changes and that potentially act at the onset of mannitol-induced leaf size reduction are largely unknown. Here, we performed a proteome and phosphoproteome analysis on growing leaf tissue of Arabidopsis thaliana plants exposed to mild mannitol-induced stress and captured the fast (within the first half hour) events associated with this stress. Based on this in-depth data analysis, 167 and 172 differentially regulated proteins and phosphorylated sites were found. We provide these data sets as a community resource and we flag differentially phosphorylated proteins with described growth-regulatory functions, but we also illustrate potential novel regulators of shoot growth.Keywords:
Proteome
Osmotic shock
Two Arabidopsis mutants atmyb123 and atkor1 were identified from the T-DNA insertion knockout mutant lines SAIL_005260 and SAIL_2_G11,respectively,and then a double mutant atmyb123/atkor1 was established by crossing method.The two mutants are lacking expression for ATMYB123 and ATKOR1 genes,respectively,which two were found to be tightly related to root development in Arabidopsis thaliana.The results obtained here showed that lack of ATMYB123 gene in expression led to a slow growth of plant rosettes and a yellow skin of seeds in Arabidopsis,while lack of ATKOR1 gene in expression had no marked effects on these two factors.Any one of the two genes ATMYB123 and ATKOR1 knockout extremely repressed the root development in Arabidopsis,especially the knockout of ATKOR1 gene,the mutant atkor1 showed only one third of length of roots as compared to wild type(WT).Interestingly,the double mutant atmyb123/atkor1 exhibited the characteristics of the single mutant atmyb123 has in plant rosette morphology and seed skins but presented intermediated root length between the two single mutants.In addition,the growth trend of roots among the three mutants had no fundamental changes when the plants were cultivated under different pH,NaCl treatments and GA concentration conditions,which imply that these three factors were not concerned in the root shortening event induced by lack of any one of ATMYB123 or/and ATKOR1 proteins in A.thaliana.These results suggest that both ATMYB123 and ATKOR1 genes participate in the root development of Arabidopsis and a specific relationship in functions exist between the two proteins,ATMYB123 and ATKOR1.The transcription factor ATMYB123 might act as a major regulator of ATKOR1 protein for participating the control of root development in Arabidopsis.
Rosette (schizont appearance)
Wild type
Gene knockout
Root hair
Lateral root
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尽管高活动性组 B (HMGB ) 蛋白质被识别了从许多在对改变环境条件的植物回答的植物种类，他们的重要性和功能的角色大部分是未知的。这里，我们调查了在对环境刺激的植物回答从黄瓜(Cucumis sativus L.) 孤立的 CsHMGB 的功能的角色。在正常生长条件下面或当使遭到了到冷应力时，在植物生长的差别都没在表示 CsHMGB 上在野类型、转基因的 Arabidopsis thaliana 之间被发现。由对比，当在高盐或脱水压力条件下面成长时，转基因的 Arabidopsis 植物与野类型的植物相比显示了延迟的萌芽。转基因的植物的萌芽被 abscisic 酸(骆驼毛的织物) 的增加推迟，暗示 CsHMGB 影响通过一个骆驼毛的织物依赖者方法的萌芽。CsHMGB 的表示影响了仅仅萌芽阶段，并且 CsHMGB 没在压力条件下面影响转基因的植物的幼苗生长。几萌芽应答的基因的抄本层次被 CsHMGB 的表示在 Arabidopsis 调制。总起来说，这些结果建议在 Arabidopsis 的 CsHMGB 的那宫外的表情调制几萌芽应答的基因的表示，并且从而在不同压力条件下面影响 Arabidopsis 植物的萌芽。
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Drought stress
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Research Article: The effects of aluminum toxicity on the protein expression of Arabidopsis thaliana
Wild type and mutant Arabidopsis thaliana plants were used to investigate the protein expression of plants exposed to elevated concentrations of aluminum. Plants were grown in a growth chamber at 25°C, 16-hour day, and 8-hour dark for eight weeks before subjecting them to different concentrations of AlCl3 solution. Time course experiments showed that Arabidopsis plants respond to aluminum toxicity by altering their protein expression. Our data indicate that aluminum toxicity caused Arabidopsis plants to express a protein in the range of 97 kD, and a medium-molecular weight protein in the range of 45 kD. Our data also indicate that responses by Arabidopsis plants to Al toxicity are not similar to responses to stresses that cause the induction of HSP70.
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作为钙传感器，种钙依赖者蛋白质 kinases (CDPK ) 在对各种各样的不能生活的压力的植物回答起重要作用。这里，我们报导 CPK28 的功能的描述，在 Arabidopsis 的 CDPK 家庭的一个成员，响应渗透的应力。cpk28 异种， loss-of-function 异种，在绿子叶展出了 NaCl 敏感、甘露糖醇敏感的显型，当 CPK28-overexpressing 植物比野类型的植物显示了更强壮的忍耐到 NaCl 和甘露糖醇压力时。染色试金的反向的 transcriptase 聚合酶链反应和 beta-glucuronidase 证明 NaCl 和甘露糖醇压力导致了 CPK28。CPK28-overexpressing 线显著地在 NaCl 和甘露糖醇压力下面相对野类型的植物和变异的植物积累了更多的脯氨酸。在叶肉房间原物的 CPK28-GFP 的短暂表示，以及表示 CPK28-GFP 的稳定的转基因的线，证明 CPK28 在血浆膜是局部性的。已知的压力应答的基因的表示层次显著地没在空异种和 overexpression 线被改变，建议 CPK28 可能调停了经由目标蛋白质的规定而非经由在抄写的水平的规定的压力反应。同时， CPK28 能 autophosphorylate。一起拿，这些数据表明了那 CPK28，一个潜在的积极管理者，在 Arabidopsis 涉及对渗透的应力的反应。
Osmotic shock
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Abstract: Polyclonal antisera against a fusion protein of β‐galactosidase and the 20 C‐terminal amino acids of the Arabidopsis thaliana sucrose carrier AtSUC2 were used to determine the cellular localization of the AtSUC2 protein. Using fluorescence‐labelling on sections from different organs of Arabidopsis the AtSUC2 protein was immunolocalized exclusively in companion cells. The presented data indicate that phloem loading in Arabidopsis may be catalyzed by the AtSUC2 sucrose carrier which transports sucrose into the companion cells. No evidence for a participation of the second Arabidopsis sucrose transporter AtSUC1 has been obtained.
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The HAP complex has been found in many eukaryotic organisms. HAP recognizes the CCAAT box present in the promoters of 30% of all eukaryotic genes. The HAP complex consists of three subunits - HAP2, HAP3 and HAP5. In this paper, we report the biological function of the AtHAP3b gene that encodes one of the HAP3 subunits in Arabidopsis. Compared with wild-type plants, hap3b-1 and hap3b-2 mutants exhibited a delayed flowering time under long-day photoperiod conditions. Moreover, the transcription levels of FT were substantially lower in the mutants than in the wild-type plants. These results imply that AtHAP3b may function in the control of flowering time by regulating the expression of FT in Arabidopsis. In a subsequent study, AtHAP3b was found to be induced by osmotic stress. Under osmotic stress conditions, the hap3b- 1 and hap3b-2 mutants flowered considerably later than the wild-type plants. These results suggest that the AtHAP3b gene plays more important roles in the control of flowering under osmotic stress in Arabidopsis.
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Transcription
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Ozet. Arabidopsis thaliana (Arabidopsis)’nin kromozom sayisinin az olmasi, bu bitkinin genetik yapisinin diger bitki turlerine gore daha kolay calisilmasina olanak saglamakta, ayrica diger bitkilerde patojenlere karsi gozlenen ana savunma mekanizmalari bu bitkide de bulunmaktadir. Bu acidan, konukcu bitkilerin patojen saldirilarina karsi savunma mekanizmalarini calisma konusunda Arabidopsis bitkisi ideal bir model sistem olusturmaktadir. Bu derlemenin amaci, patojenlere karsi bitki savunma mekanizmasinin genetigini anlamada Arabidopsis’in bir sablon olarak nasil kullanildigini, bitki ekotiplerinin patojen izolatlarina karsi gosterdigi farkliliklari (duyarlilik ve dayaniklilik) ortaya koyacak inceleme metodlarini aciklamaktadir. Anahtar Kelimeler: Arabidopsis thaliana, Peronospora parasitica, hastaliklara dayaniklilik, mutant bitkiler A Model Plant In Host-Pathogen Interaction: Arabidopsis thaliana Abstract. Because of its small genome, genetic studies are performed easier in Arabidopsis thaliana (Arabidopsis) compared to other plants and it exhibits all of the major kinds of defense responses described in other plants. Thus, Arabidopsis provides to be an ideal model system for studies of host defense responses to pathogen attack. This review details methods to isolate and utilize phytopathogens useful as probes in understanding the genetics of plant defense responses in Arabidopsis. As well, it will be defined how screening systems can be set up to detirmine differential responses (resistance and susceptibility) of Arabidopsis ecotypes to pathogen isolates. Key Words: Arabidopsis thaliana, Peronospora parasitica, disease resistance, mutant plants
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Osmotic shock
Osmotic pressure
Osmoregulation
Esterase
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The article presents a study on the mutual influence of mutant genes in productivity of Arabidopsis plant ( Arabidopsis thaliana (L.) Heynh.), in different growing models of plant communities. It is shown that under the joint cultivation of five genetically pure lines of Arabidopsis is occurring mutual inhibition and aid, which leads decreasing or increasing of seed production plants.
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[Objective]The aim was to study the action mode of high mobility group B(HMGB)proteins of higher eukaryotic cells in the transcriptional regulation of plant stress responses.[Method]The At2G33450 gene encoding AT2G34450 protein in Arabidopsis thaliana was cloned.Then,binary vectors carrying the gene were transformed into Arabidopsis thaliana and the over-expression strains were selected to detect the influences of environmental stimuli on transgenic Arabidopsis.[Result]Under salt or drought stress,the transgenic Arabidopsis plants which over-expressed At2G33450 displayed retarded germination and subsequent growth compared with wild-type plants.[Conclusion]At2G33450 protein of HMGB protein family played an important role in the growth and development of Arabidopsis plants under various stress conditions.
Heterologous expression
Heterologous
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