植物生长发育和逆境响应中SR-like蛋白的研究进展

同一基因pre-mRNA经可变剪接(alternative splicing, AS)后能够产生不同的转录本,使得编码的蛋白在细胞中的定位、稳定性和翻译后修饰的功能发生改变,进而增强应答发育及环境胁迫的能力,富含丝氨酸-精氨酸蛋白(serine/arginine-rich proteins, SR proteins/SR)是决定可变剪接效率和准确性的一个重要剪接因子家族。该文在简要介绍SR蛋白概念、分类的基础上,首次系统综述了植物特有的SR蛋白亚家族SR-like(SR45/45a)结构特点、成员构成、亚细胞定位和转录调控功能,尤其是对于非生物胁迫应答过程中相关基因可变剪接的调控机制进行了阐述,并展望了未来植物SR-like可能的前景方向和研究内容。     

聚乙二醇丙烯酸酯的合成与表征

以聚乙二醇-400（PEG400）与丙烯酸直接缩合反应,在不加有毒带水剂的条件下合成了丙烯酸聚乙二醇酯（PEGA）。通过正交实验确定酯化反应的最佳条件：丙烯酸／PEG400的摩尔比为2．0：1．0,反应温度是110℃,阻聚剂对苯二酚为0．4％（以醇酸总质量计）,反应时间为6小时,催化剂对甲苯磺酸为0．8％（以醇酸总质量计）,产率为76．7％。产品结构经IR和1HNMR表征,证明是所需的产物。     

Cell Death and Growth Recovery of Barley after Transient Salt Stress

3 H-thymidine) in old roots were partially recycled for new tissue formation. This result indicates that cell death may have some physiological roles under transient salt stress. Received 11 January 2000/ Accepted in revised form 29 June 2000     

Response of Split-Root Sour Orange Seedlings to NaCl and Polyethylene Glycol Stresses

Soil water cotent and salinity levels are seldom uniform inthe field, particularly with the use of micro-irrigation systemsthat may water only a portion of the root zone. For studyingnon-uniform salinity, a split-root experiment was designed toevaluate growth and water relations when half of the root systemof sour orange (Citrus aurantium) seedlings was stressed withsodium chloride (NaCl) or polyethylene glycol (PEG). This studyalso determined if non-stressed portions of the root systemcompensated for the decrease in water uptake by the stressedportions. One or both halves of the root system were treated for fourmonths with nutrient solution adjusted with NaCl or PEG to osmoticpotentials of –0.10, –0.20, or –0.35 MPa.Shoot dry weight was reduced by only 9% when half of the rootsystem was irrigated with saline solution at –0.10 MPa,but with both halves of the root system at –0.10 MPa,shoot and root dry weights were reduced as much as 45%. Similarly,leaf water and osmotic potentials were also more disturbed underuniform salinity than under non-uniform salinity conditions. Plant growth, leaf water potential, osmotic potential, stomatalconductance, and evapotranspiration decreased with increasingNaCl and PEG concentrations in the nutrient solution. Turgorpotential and leaf thickness increased in response to NaCl treatments.Microscopic examination showed that the increase in leaf thicknesswas due to the development of larger cells in the spongy mesophyll. Shoot growth did not correlate with the average osmotic potentialof the two root halves. Seedlings with one stressed half-rootsystem had shoot dry weight and leaf water potential valuescloser to those of the non-stressed control than to those withthe completely stressed root system. Key words: Non-uniform salinity, water relations, citrus     

Water Uptake and Protein Synthesis in Germinating Wheat Embryos under the Osmotic Stress of Polyethylene Glycol

The effect of osmotic stress on wheat-seed germination was testedby imbibition in aqueous polyethylene glycol solutions at differentconcentrations. The experiments were designed to allow blockingand the subsequent recovery of germination by 12 h or 24 h pre-imbibitionof seeds in osmoticum, followed by transfer to water. Seedswere alternatively presoaked in water for 12 or 24 h, then transferredto polyethylene-glycol solutions to study the induced blockingof germination. Water content and [³H]leucine incorporationinto embryo tissues (as a measure of in vivo protein synthesis)were determined over a 48-h imbibition period. A close relationshipwas established overall between hydration status and proteinsynthesis rate. Osmotic stress seems to have a strong influenceupon the quantitative synthesis of proteins, suggesting thatthis biochemical activity is associated with the regulationof the germination process. Triticum durum, embryo, osmotic stress, water uptake, protein synthesis     

谷子幼苗对土壤铅、铬的生长响应及吸收积累的差异性

采用盆栽土培试验,研究了谷子幼苗对土壤中不同含量铅（Pb）、铬（Cr）的生长响应和吸收积累的差异性。结果表明,在所试浓度（50～800 mg·kg^-1）范围内,Pb、Cr在谷子幼苗地上部和地下部的积累量存在较大差异,幼苗重金属的吸收富集和转运系数均为Pb大于Cr、Pb、Cr胁迫对幼苗生物量的影响表现为低浓度的促进和高浓度的抑制作用,但Cr对生物量的影响比Pb更强。相同处理条件下,幼苗茎叶中可溶性蛋白质、DNA含量和增色效应对Pb、Cr的响应也有明显差别,Cr对幼苗的生理毒性和DNA损伤效应的作用强度大于Pb。     

A Theory for the Displacement of Proteins and Viruses with Polyethylene Glycol

The displacement action of polyethylene glycol of different molecular weights may be linked to the ability of the polymers to form coiled particles in solution. From conclusions drawn from their sedimentating properties in centrifugal fields the polyethylene glycols of low molecular weights, as expected, are les randomly coiled than those of higher molecular weight. It is suggested that protein molecules have the ability to diffuse into the coils of the polyethylene glycol from which they are excluded when the random coiling increases with increasing polymer concentration. From considerations based on the interaction of the polymer filament with the displaced particle the distribution of the substance between the coils and the intermolecular spaces may be predicted semi-quantitatively.     

Plant Tandem CCCH Zinc Finger Proteins Interact with ABA,Drought, and Stress Response Regulators in Processing-Bodies and Stress Granules

Although multiple lines of evidence have indicated that Arabidopsis thaliana Tandem CCCH Zinc Finger proteins, AtTZF4, 5 and 6 are involved in ABA, GA and phytochrome mediated seed germination responses, the interacting proteins involved in these processes are unknown. Using yeast two-hybrid screens, we have identified 35 putative AtTZF5 interacting protein partners. Among them, Mediator of ABA-Regulated Dormancy 1 (MARD1) is highly expressed in seeds and involved in ABA signal transduction, while Responsive to Dehydration 21A (RD21A) is a well-documented stress responsive protein. Co-immunoprecipitation (Co-IP) and bimolecular fluorescence complementation (BiFC) assays were used to confirm that AtTZF5 can interact with MARD1 and RD21A in plant cells, and the interaction is mediated through TZF motif. In addition, AtTZF4 and 6 could also interact with MARD1 and RD21A in Y-2-H and BiFC assay, respectively. The protein-protein interactions apparently take place in processing bodies (PBs) and stress granules (SGs), because AtTZF5, MARD1 and RD21A could interact and co-localize with each other and they all can co-localize with the same PB and SG markers in plant cells.     

逆境相关植物锌指蛋白的研究进展

锌是植物必需的营养元素,锌指蛋白因其具有指状结构特征且能结合Zn2 而得名,植物锌指蛋白包含特有的QALGGH保守结构,可能涉及调控植物特有的生物学功能。人们已经从拟南芥(Arabidopsis thaliana)、矮牵牛(Petunia hybrida Vilm)、水稻(Oryza sativa)、大豆(Glycine max)、棉花(Gossypium hirsutum)等植物中克隆了许多编码锌指蛋白的基因,并对其结构及功能进行了研究。利用转基因技术,将一些与逆境胁迫相关的锌指蛋白基因在目标植物中过量表达后,能对植物起到增强抗逆性的作用,说明锌指蛋白在增强植物逆境抗性方面有着广阔的应用前景。     

NaCl 胁迫对盐芥和拟南芥光合作用的影响

本研究检测了与盐芥(Ghellungiella halophila)和拟南芥(Arabidopsis thaliana)光合作用相关的叶绿素、净光合速率(photosynthetic rate, Pn)、气孔导度(stomatal conductance, Gs)、胞间隙CO2浓度以及叶绿素荧光参数等指标, 观察到随着NaCl浓度逐渐增加, 盐芥的叶绿素a/b值(Chl a/Chl b)、类胡萝卜素/总叶绿素值(Car/Chl)显著高于拟南芥, 且二比值变化幅度较小并保持较高水平。盐胁迫下拟南芥净光合速率下降、气孔导度下降和胞间CO2浓度减小。气孔因素是引起拟南芥光合能力下降的主要因素。叶绿素荧光参数的变化表明, 50-200 mmol.L-1 NaCl降低拟南芥叶绿体对光能的吸收能力, 而且降低叶绿体的光化学活性, 使电子传递速率和光能转化效率大幅度下降,造成光能转化为化学能的过程受阻,进一步加剧了光合放氧和碳同化能力的降低。而50-200 mmol.L-1 NaCl 胁迫没有使盐芥的光合作用受到不良影响。     

NaCI胁迫对盐芥和拟南芥光合作用的影响

本研究检测了与盐芥(Ghellungiella halophila)和拟南芥(Arabidopsis thaliana)光合作用相关的叶绿素、净光合速率(photosynthetic rate,Pn)、气孔导度(stomatal conductance,Gs)、胞间隙CO2浓度以及叶绿素荧光参数等指标,观察到随着NaCl浓度逐渐增加,盐芥的叶绿素a／b值(Chl a／Chl b)、类胡萝卜素／总叶绿素值(Car／Chl)显著高于拟南芥,且二比值变化幅度较小并保持较高水平。盐胁迫下拟南芥净光合速率下降、气孔导度下降和胞间CO2浓度减小。气孔因素是引起拟南芥光合能力下降的主要因素。叶绿素荧光参数的变化表明,50-200 mmol·L-1NaCl降低拟南芥叶绿体对光能的吸收能力,而且降低叶绿体的光化学活性,使电子传递速率和光能转化效率大幅度下降,造成光能转化为化学能的过程受阻,进一步加剧了光合放氧和碳同化能力的降低。而50-200 mmol·L-1NaCl胁迫没有使盐芥的光合作用受到不良影响。     

A Full-Length Dof1 Transcription Factor of Finger Millet and its Response to a Circadian Cycle

DOF1 (DNA binding with one finger) plays an important role in regulating C/N metabolism in cereals. In order to validate its role in the regulation of nitrogen use efficiency (NUE) and photosynthetic efficiency in finger millet, 5′–3′ RACE PCR was performed to obtain and characterize full-length Dof1 genes of high and low grain protein finger millet genotypes. The full-length DOF1 ORFs were both 1,284 nt long and were 98.8 % similar over 427 amino acids containing the characteristic Dof domain. Comparison of both the EcDof1 protein sequences with the Dof1 of other cereals revealed high sequence similarity to the Dof1 of rice. Southern hybridization carried out using the probe developed from the region encoding the highly variable C-terminal region of EcDof1 showed the presence of four copies of the DOF1 gene in finger millet, which might explain the high NUE and photosynthetic performance of finger millet. Since the genes involved in C/N metabolism are regulated diurnally and play crucial roles in determining grain protein content during grain filling, the diurnal expression of EcDOF1 was assessed in two finger millet genotypes (GE 3885 and GE 1437) with differing grain protein content (13.8 % and 6.15 % respectively). It was found that EcDOF1 exhibited diurnal regulation and peak differential pattern expression with early phasing in GE3885 and late phasing in GE1437. Differential expression of DOF1 might alter the regulation of genes involved in C/N metabolism affecting grain protein composition of finger millet genotypes.     

Coordinate Gene Response to Salt Stress in Lophopyrum elongatum

Lophopyrum elongatum is a highly salt-tolerant relative of wheat. A previous study showed that the abundance of a number of mRNA species is enhanced or reduced in the roots of the L. elongatum × Triticum aestivum amphiploid by salt stress. Eleven genes with enhanced expression in the roots of salt-stressed L. elongatum plants have been cloned as cDNAs. The clones were used as probes to characterize temporal expression of these genes in roots after initiation of salt (250 mm NaCl) stress. All 11 genes are induced within 2 h after exposure to 250 mm NaCl and reached peak expression after 6 h. The decline of gene expression distinguished two groups, one in which mRNA concentrations returned to basal levels by 24 h and the other in which this occurred between 3 and 7 d. One of the 11 clones was found to be homologous to a multigene family of abscisic acid-induced genes, rab and dhn, identified in other species. We suggest that the coordinate expression of this large number of genes reflects the existence of a highly specific early response to salt stress. We refer to this response as the “early salt stress response.”     

聚乙二醇在植物渗透胁迫生理研究中的应用

介绍聚乙二醇(PEG)在调节培养液和固体培养基水势以及在水分胁迫和盐胁迫研究中的应用及应注意的问题,并介绍PEG溶液水势的测定方法和水势计算公式.     

植物耐盐基因工程研究进展

盐害是影响植物生长和作物产量的主要因素之一。用于提高植物耐盐性的基因工程方法很多,最常见的就是在植物中过量表达抗盐相关的功能基因,包括植物信号传导蛋白基因、植物离子通道蛋白基因和合成小分子渗透剂的酶基因等。归纳了近年来植物耐盐基因工程的研究进展,并展望了植物耐盐基因工程的研究前景。     

Growth, Gas Exchange, Abscisic Acid, and Calmodulin Response to Salt Stress in Three Poplars

In the present study, we Investigated the effects of Increaslng sallnlty on growth, gas exchange, absclslc acld （ABA）, calmodulln （CAM）, and the relevance to salt tolerance In seedllngs of Populus euphratlca Ollv. and cutUngs of P. ＂pupularls 35-44＂ （P. popularls） and P. x euramerlcana cv. 1-214 （P. cv. Itallca）. The relatlve growth rates of shoot helght （RGR,） for P. cv. Itallca and P. popularls were severely reduced by Increaslng salt stress, whereas the growth reductlon was relatlvely less in P. euphratica. Slmllarly, P. euphratlca malntalned hlgher net photosynthetlc rates （Pn） and unlt transplration rotes （TRN） than P. cv. Itallca and P. popularls under condltlons of hlgher sallnlty. Sallnity caused a slgnlficant increase In leaf ABA and CaM In the three genotypes after the onset of stress, but NaCl-induced ABA and CaM accumulatlon was more pronounced In P. euphratlca, suggeeUng that P. euphratlca plants are more sensitlve in sensing soil salinlty than the other two poplars. Furthermore, P. euphratica maintained relatively higher ABA and CaM concentrations under conditions of high salinity. The higher capacity to synthesize stress signals, namely ABA and CaM, In P. euphratica and the contrlbuUon of this to the salt resistance of P. euphratica are discussed.