Pathogenesis-related proteins and their genes in cereals

Pathogenesis-related proteins (PR-proteins) are induced in plants in response to attack by microbial or insect pests. They have been classified into several groups (PR-1 through PR-14 at present) based on their amino acid sequences and biochemical functions. Many of these proteins that have been purified from infected plants or seed extracts possess antifungal or insecticidal activity. Genes and cDNA clones for all classes of PR-proteins have been isolated from a variety of cereals. Some of these genes/cDNAs have been used to transform cereals. This review presents a summary of the PR-proteins and their genes characterized from rice, wheat, barley, sorghum and maize. Efforts to improve disease or insect resistance of these cereal plants by genetic engineering using genes for PR-proteins also are discussed. In many cases, the expression of the PR-proteins either singly or in combination appears to improve resistance to fungi or insects. In addition, chromosomal location of the PR-protein genes indicates that members of the same family of PR-protein genes or sometimes even several families of PR-protein genes often are clustered in the cereal genome, suggesting coordinate regulation. Some of these PR-protein genes map closely to quantitative traits loci. Some concerns regarding the use of genes encoding PR-proteins for genetic modification of cereals also are addressed.     

Pathogenesis-related proteins are developmentally regulated in tobacco flowers

The accumulation of pathogenesis-related proteins (PR) in tobacco leaves has been casually related to pathogen and specific physiological stresses. The known enzymatic function of some of these proteins is potentially antimicrobial. By using antibodies specific to three classes of pathogenesis-related proteins, we examined tobacco plants during their normal growth. The pathogenesis-related proteins accumulated during the normal development of the tobacco flower. The PR-1 class of proteins (biological function unknown) is located in sepal tissue. PR-P, Q polypeptides are endochitinases and are present in pedicels, sepals, anthers, and ovaries. A glycoprotein serologically related to the PR-2,N,O class is a (1,3)-beta-glucanase and is present in pistils. Differential appearance during flower development, in situ localization, and post-translational processing of floral pathogenesis-related proteins point to a hitherto unsuspected function these classes of pathogenesis-related proteins play in the normal process of flowering and reproductive physiology.     

Pathogenesis-related proteins protect extrafloral nectar from microbial infestation

Plants in more than 300 genera produce extrafloral nectar (EFN) to attract carnivores as a means of indirect defence against herbivores. As EFN is secreted at nectaries that are not physically protected from the environment, and contains carbohydrates and amino acids, EFN must be protected from infestation by micro-organisms. We investigated the proteins and anti-microbial activity in the EFN of two Central American Acacia myrmecophytes ( A. cornigera and A. hindsii ) and two related non-myrmecophytes ( A. farnesiana and Prosopis juliflora ). Acacia myrmecophytes secrete EFN constitutively at high rates to nourish the ants inhabiting these plants as symbiotic mutualists, while non-myrmecophytes secrete EFN only in response to herbivore damage to attract non-symbiotic ants. Thus, the quality and anti-microbial protection of the EFN secreted by these two types of plants were likely to differ. Indeed, myrmecophyte EFN contained significantly more proteins than the EFN of non-myrmecophytes, and was protected effectively from microbial infestation. We found activity for three classes of pathogenesis-related (PR) enzymes: chitinase, β-1,3-glucanase and peroxidase. Chitinases and β-1,3-glucanases were significantly more active in myrmecophyte EFN, and chitinase at the concentrations found in myrmecophyte EFN significantly inhibited yeast growth. Of the 52 proteins found in A. cornigera EFN, 28 were annotated using nanoLC-MS/MS data, indicating that chitinases and glucanases contribute more than 50% of the total protein content in the EFN of this myrmecophyte. Our study demonstrates that PR enzymes play an important role in protecting EFN from microbial infestation.     

Pathogenesis-related genes and proteins in forest tree species

Trees occupy more than 30% of the land biosphere. They are important from both ecological and environmental standpoints and provide some of the most valuable commodities in the world economy. The perennial nature and size of trees are the critical determinants of their survival in response to biotic and abiotic stresses. The identification of the defense pathways at biochemical and genetic levels in tree pathosystems are beginning to be addressed. The basic physiological and biochemical mechanisms in woody perennials in response to pathogen is homologous to the model annual crop like Arabidopsis, but their secondary metabolic processes and ecological survival strategies are likely to be divergent from their annual counterparts. The limited domestication in tree species makes its molecular mechanisms less comparable to the highly pedigreed crop species. Recent reports have highlighted that the possible difference in genetic programs responding to invasive pathogens between annuals and perennials could be the spatial and temporal pattern of gene regulation. Several reviews on pathogen defense with reference to crop species are available, while similar reports from the tree species are limited to few commercially important species like Populus, Pinus, Picea, Eucalyptus, Castanea, and Pseudotsuga. This paper reviews the present status of pathogenesis-related genes and proteins from tree species with emphasis on the resistant genes and the proteins induced during systemic acquired resistance and highlights the ecological and evolutionary significance of defense-related genes from tree species.     

Pathogenesis-related proteins in somatic hybrid rice induced by bacterial blight

Rice bacterial blight, caused by Xanthomonasoryzae pv. Oryzae (Xoo), is one of the most serious rice diseases worldwide. The bacterial blight resistance trait from Oryza meyeriana, a wild rice species, was introduced into an elite japonica rice cultivar using asymmetric somatic hybridization. This study was carried out with the intention of understanding the molecular mechanism of incompatible interaction between Xoo and the stable somatic hybrids by using proteomic analyses. Proteins were extracted from leaves at 24, 48, and 72 h after Xoo inoculation and separated by 2-DE. A total of 77 protein spots changed their intensities significantly (p<0.05) by more than 1.5-fold at least at one time point. Sixty-four protein spots were successfully identified by MS analysis. Among them, 51 were known to be involved in photosynthesis. Up-regulation of Rubisco large subunit (RcbL) small fragments and down-regulation of RcbL big fragments indicated that intact RcbL and RcbL big fragments degraded following Xoo attack, which was further confirmed by Western blot analysis. The differential expression of proteins related to signal transduction, antioxidant defense, photosynthesis, metabolism, and protein turnover during the Xoo infection, suggests the existence of a complex regulatory network in the somatic hybrid rice that increases resistance toward Xoo infection and damage.     

Pathogenesis-related proteins in plants and tissues ofNicotiana tabacum transformed byAgrobacterium tumefaciens

Large amounts of pathogenesis-related (PR) proteins were found inNicotiana tabacum crown gall tissue, following transformation of normal tobacco cells withAgrobacterium tumefaciens. In contrast, PR proteins were not detected in leaves of grafted plants that had been recovered from crown gall tissue even though these plants were still transformed as shown by their inability to form roots and ability to produce octopine. No difference was observed in susceptibility to virus infection between untransformed and transformed plants grafted onto identical rootstocks. The results are discussed in relation to physiological factors controlling PR protein induction and virus resistance.     

Pathogenesis-related proteins of tomato : p-69 as an alkaline endoproteinase

An endoproteinase induced by citrus exocortis viroid has been purified from tomato (Lycopersicon esculentum Mill, cv “Rutgers”) leaves. The proteinase corresponds to one of the major pathogenesis-related proteins of tomato plants and was designated proteinase P-69 as it has a molecular weight of 69,000 to 70,000. The proteinase was purified in four steps: (NH₄)₂SO₄ fractionation, chromatography on Bio-Gel P-60, DEAE-Sepharose chromatography, and casein-Sepharose affinity chromatography. The proteinase had a pH optimum of 8.5 to 9.0 when assayed with either fluorescein thiocarbamoyl derivative (FTC)-casein or FTC-ribulose 1,5-bisphosphate carboxylase/oxygenase as substrates. The proteinase activity was inhibited by pCMB and strongly activated by calcium and magnesium ions as well as by DTT. When analyzed by electrofocusing, the activity showed a pI around 9.0.     

Pathogenesis-related proteins and polyamines in a developmental mutant of tomato, epinastic

The polyamine level and the accumulation of pathogenesis-related (PR) proteins were studied in the ethylene overproducing Epinastic (Epi) tomato (Lycopersicon esculentum Mill.) mutant, as compared with its parent, cv VFN8. Neither a decreased putrescine level nor an enhanced production of PR proteins were detected in Epi, contrary to what could be expected from our previous studies (JM Bellés, J Carbonell, V Conejero [1991] Plant Physiol 96: 1053-1059). However, treatment with the ethylene-releasing compound 2-chloroethylphosphonic acid (ethephon) or silver nitrate at high doses induced a decrease in putrescine content and an enhancing of the synthesis of PR proteins in Epi as ascertained by immunoblot analysis using antisera raised against Rutgers tomato PR proteins.     

Pathogenesis-related genes of entomopathogenic fungi

All living things on Earth experience various diseases such as those caused by viruses, bacteria, and fungi. Insects are no exception to this rule, and fungi that cause disease in insects are called entomopathogenic fungi. These fungi have been developed as microbial insecticides and are used to control various pests. Generally, the mode of action of entomopathogenic fungi is divided into the attachment of conidia, germination, penetration, growth, and generation of secondary infectious conidia. In each of these steps, that entomopathogenic fungi use genes in a complex manner (specific or diverse) has been shown by gene knock-out and RNA-sequencing analysis. In this review, the information mechanism of entomopathogenic fungi was divided into six steps: (1) attachment of conidia to host, (2) germination and appressorium, (3) penetration, (4) fungal growth in hemolymph, (5) conidia production on host, and (6) transmission and dispersal. The strategy used by the fungi in each step was described at the genetic level. In addition, an approach for studying the mode of action of the fungi is presented.     

高等植物病原相关蛋白

在过去的三十年中,人们对诱导系统性抗性——这一普遍存在于高等植物抗病过程中的现象——进行了深入研究。被真菌、细菌或病毒侵染后,植物表现出广泛的、长时间的系统性抗性。在这一过程中,植物细胞壁组成成分发生改变,表达各种病原相关蛋白(PR蛋白),并合成多种植物抗毒素。本文就主要的PR蛋白家族的结构和功能特性,PR蛋白的发现和分类,及PR蛋白的应用作一综述。     

胶质瘤发病相关蛋白1与肿瘤

胶质瘤发病相关蛋白1（glioma pathogenesis-related protein 1,GLIPR1）隶属于CAP蛋白超家族,由1个指导分泌的信号肽、1个保守的富含半胱氨酸的结构域及1个跨膜结构域组成CAP。近年研究表明,胶质瘤发病相关蛋白1启动子甲基化水平决定了其在胶质瘤和黑色素瘤中的表达上调。而在前列腺癌、肺癌、骨肉瘤以及白血病等肿瘤中表达下调。在不同类型的肿瘤中,GLIPR1通过不同的信号通路参与调控肿瘤的发展进程。本文针对胶质瘤发病相关蛋白1参与调控多种肿瘤发生及发展的机制进行综述,以期为上述肿瘤的诊断与治疗提供重要的线索。     

植物病原相关蛋白研究进展

植物在受到真菌、细菌、病毒侵染或意外的伤害时体内会产生新的蛋白质,称作病原相关蛋白(pathogenesisrelatedprotein简称PR蛋白)。PR蛋白的产生与植物对病原物的抵御直接相关,本文就PR蛋白的诱导表达、调控和作用机理方面的最新研究进展作一综述。     

Pathogenesis-related proteins in lima bean leaves infected with tobacco ringspot virus and their distribution within and around local lesions

Tobacco ringspot virus (TRSV) induces circular, darkbrown local lesions on primary leaves of lima bean (Phaseolus lunatus cv Nemagreen) with a concomitant production of three basic and three acidic pathogenesisrelated (PR) proteins. The three basic proteins are: a 21 kDa protein related serologically to Pinto bean PR-4d and tobacco PR-5 proteins; a 36 kDa glucanase that is related to tobacco PR-2; and, a 31 kDa chitinase related serologically to ethylene-induced bean chitinase. The three acidic 18 kDa lima bean PR proteins are serologically similar and probably are charged isomers of the same protein. The 21 kDa basic protein and the 18 kDa acidic protein accumulated preferentially at the lesion center while the 31 kDa chitinase and TRSV were distributed evenly throughout the necrotic area. In green tissue immediately surrounding a lesion, the amounts of PR proteins were comparable to or lower than those in the necrotic area, and virions were not detected. This mode of spatial distribution indicates that lima bean PR proteins are not involved in TRSV localization, and is consistent with other observations that PR proteins play no direct role in restricting viral spread.     

Pathogenesis-related proteins induced by agroinoculation-associated cell wall weakening can be obviated by spray-on inoculation or mannitol ex vivo culture

Agroinoculation transient expression systems are commonly accompanied with elevated pathogenesis-related (PR) protein production and leaf necrosis. We identified the major PR proteins in Nicotiana benthamiana in response to agroinoculation and determined that their occurrence was mainly due to agrobacterium infection and the method of inoculation, rather than due to viral vectors overexpressing foreign proteins. A spray-on inoculation method was optimized in this research and used to obviate the leaf necrosis and PR proteins induced by agrobacterium. Subsequently, this method also increased the yield and purity of the protein-of-interest. A further investigation of PR protein induction by a necrosis-inducing protein, Jun a 1, suggested that the plant pathogenic response was related to biochemical integrity of plant cell wall, which was also confirmed by an osmo-stabilizing mannitol ex vivo culture experiment. These findings provide insight into the response of plants to agroinoculation and suggested a connection between cell wall weakening and PR protein elicitation.     

植物病程相关蛋白及其在烟草中的研究进展

植物在受到病原物侵染时,会产生一系列抗性反应,病程相关蛋白是其中参与抗病性的重要物质,能够被病原物诱导产生并在植物体内积累,对于诱导植物系统抗性,阻止病原物侵染具有重要作用。对植物病程相关蛋白的性质、诱导因素、分类和功能进行综述,并概述病程相关蛋白与烟草系统抗性的紧密联系以及烟草病程相关蛋白基因在增强系统抗性中的应用,为烟草抗病育种和病虫害防治提供了理论。     

Light-induced Changes in Expression of Pathogenesis-related Anionic Peroxidase in Cucumber Seedlings

The effect of irradiance on the expression of the Cucumis sativus pathogenesis-related (srPRX) anionic peroxidase was studied in germinating seeds at the period when seedlings start to be photosynthetically active. The diversity in the expression patterns of srPRX was observed in both dark- and light-grown seedlings using activity staining and immunoblotting: beside the three srPRX isoenzymes also other three, serologically unrelated, peroxidase isoforms were accumulated in dark-grown seedlings and one in light-grown seedlings. Furthermore, in light-grown seedlings, it was observed a marked difference in the expression of particular srPRX isoenzymes at higher irradiance (up to 260 W m-2, 400 - 700 nm) in comparison to low irradiance. By tissue printing on nitrocellulose paper it was demonstrated that irradiance during germination induced changes in the temporal and spatial distribution of the srPRX.     

病程相关基因启动子片段与GUS的融合基因在拟南芥中的表达

PR1是拟南芥(Arabidopsisis thaliana L.)系统获得抗性的一个标志基因.利用PCR技术,从拟南芥中扩增并克隆了PR1基因的启动子片段.将该启动子片段与GUS报告基因拼接,构建成含有PR1-GUS融合基因的重组表达质粒.经根癌农杆菌介导转化,得到了转基因的拟南芥植株.用已知的系统获得抗性激活剂处理转基因植物,检测到GUS活性.因此,这一转基因体系可以作为一种简便、灵敏的实验体系以筛选激活植物系统获得抗性的化合物.