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1.
David J. McNally Kirstin V. Wurms Caroline Labb Richard R. Blanger 《Physiological and Molecular Plant Pathology》2003,63(6):293-303
Leaf tissue harvested from cucumber plants (Cucumis sativus L.) expressing induced resistance against the powdery mildew fungus Podosphaera xanthii (syn. Sphaerotheca fuliginea, Castagne; Braun and Shishkoff) was extracted and analyzed for phytoalexin compounds. Fluorescence microscopy was then used to observe the production of these compounds in planta, and laser scanning confocal microscopy observations were made to locate the subcellular sites of phytoalexin accumulation. Phytochemical analyses and fluorescence microscopy observations revealed the production of autofluorescent C-glycosyl flavonoid phytoalexins within the epidermal tissues of disease-resistant plants undergoing fungal ingress. Phytoalexin production was triggered by the combination of an eliciting/inoculation treatment, and tissue autofluorescence of color characteristic of the phytoalexins reached a maximum 48 h after elicitation prior to subsiding following the collapse of the pathogen. After a second eliciting treatment, disease-resistant plants produced phytoalexins more rapidly in response to fungal challenge. At the cellular level, autofluorescent C-glycosyl flavonoid phytoalexins were observed associated with the plasma membrane of infected epidermal cells immediately following elicitation. In the hours that preceded the collapse of conidial chains, phytoalexins accumulated inside the haustorial complexes of the pathogen within the epidermal cells of disease-resistant plants. Taken together, the results of this study show the timely synthesis of C-glycosyl flavonoid phytoalexins at precise subcellular locations as a key defense reaction used by cucumber to create incompatible interactions with powdery mildew. 相似文献
2.
《Physiological and Molecular Plant Pathology》2007,70(1-3):38-48
Flavonoids are a group of secondary plant metabolites important for plant growth and development, and thus the regulation of their biosynthesis is of special interest. We used a transgenic approach for flavonoid content manipulation. The multigene construct contained the cDNAs for chalcone synthase (CHS), and chalcone isomerase (CHI) and dihydroflavonol reductase (DFR) were prepared. Following flax plants transformation, the levels of the products of the enzyme overproduction were assessed in leaves and seeds. The simultaneous expression of genes resulted in a significant increase in the levels of flavanones, flavones, flavonols and anthocyanins, suggesting those three overproducing enzymes efficiently control the flavonoid route of the phenylpropanoid pathway.The increase in the flavonoid content in the transgenic flax plants might be the reason for observed, enhanced antioxidant capacity of those plants. The increased antioxidative properties of transgenic plants lead to improved resistance to Fusarium, the main pathogen of flax.The changes in phenylpropanoids accumulation in transgenic plants affect cell wall carbohydrate content. Immunochemical studies revealed significant increase in carbohydrates, constituents of pectin and hemicellulose. Since pectins contribute to flax stem retting, the compounds increase might affect fibre production. An increase in pectin and hemicellulose content leads to enhanced disease resistance of those plants. 相似文献
3.
4-Coumarate:CoA ligase (4CL, EC 6.2.1.12) exists only in plants and plays an important role in the phenylpropanoid pathway. Identification of inhibitors targeting 4CL provides a novel approach for developing effective plant growth inhibitors (PGIs). The full-length gene of tobacco 4CL (Nt4CL1) was cloned and expressed in Escherichia coli Cast & Chalm. The recombinant 4CL protein was extracted and purified by several purification steps including gel-filtration and anion-exchange chromatography. 4CL activity assay was miniaturized and optimized using a 96-well microplate and a reader. Among 28 existing herbicides, propanil and swep strongly inhibited in vitro 4CL enzyme activity, and they were selected for further studies. The process of this assay can be developed into a high-throughput screening system of PGI targeting 4CL in the phenylpropanoid pathway. 相似文献
4.
4-Coumarate:CoA ligase (4CL, EC 6.2.1.12) in the phenylpropanoid pathway in plants has attracted interest as a novel target for developing effective plant growth inhibitors (PGIs). In a previous study in which the 4CL inhibitory activity of 28 existing herbicides was investigated using an optimized in vitro screening assay, 4CL activity was found to be strongly inhibited by propanil and swep at 100 microM. Here, further experimental evidence is provided to substantiate the previous result. Using 4-coumaric acid as substrate, tobacco 4CL activity was inhibited by propanil or swep in a concentration-dependent manner, with 50% inhibition concentrations (I(50)) of 39.6 and 6 microM respectively. These herbicides also exhibited uncompetitive inhibition towards 4-coumaric acid. Furthermore, 4CLs from several plant species were inhibited by the herbicides within a range from 1 to 50 microM. It is proposed that these herbicides have another site of action as a result of the inhibition of 4CL in the phenylpropanoid pathway, and this enzyme represents a new target site for the development of PGI. 相似文献
5.
The 4-coumarate: coenzyme A ligase (4CL) is one of the key enzymes in the biosynthesis of lignin monomers. It has been demonstrated that the 4CL is a new potential target site for developing effective plant growth inhibitors. Although previous studies demonstrate that chalcone and naringenin differentially suppress the growth of several annual plant species, we show here that the compounds can inhibit the 4CL enzyme activity in the plants. The enzyme was extracted and partially purified from the leaf tissues of two tolerant plants (wheat and soybean) and three susceptible plants (tomato, barnyard grass, and common chickweed). A maximal 29-fold purification of the enzyme, with a yield of 32% (tomato), was achieved by a six-step procedure, including anion-exchange column chromatography. Naringenin strongly inhibited the 4CL specific activity in wheat, soybean and barnyard grass, whereas chalcone showed the highest inhibitory effect in common chickweed. A good correlation was observed between the level of growth suppression by the compounds and the total 4CL amount in the plants. These results suggest that the inhibitor treatment at the same concentration could not inactivate the entire 4CL enzyme produced in the tolerant plants. Taken together, these results highlight the possibility of the 4CL as a new action site of growth suppression. 相似文献
6.
为了明确壳聚糖(chitosan,CTS)对黄瓜幼苗抗灰霉病的诱导作用,采用根际注射结合叶面喷洒的方法,测定了灰霉病菌侵染下幼苗植株病情指数、防御酶活性和抗病相关物质含量等生理指标。结果表明:CTS降低了黄瓜幼苗的病情指数,最高幅度达39.4%,提高了苯丙氨酸解氨酶(PAL)、过氧化物酶(POD)和多酚氧化酶(PPO)活性,增加了总酚、类黄酮和木质素含量。说明CTS能诱导黄瓜幼苗对灰霉病的抗性,且最佳处理浓度为200mg·L-1。 相似文献
7.
链格孢菌Alternaria alternata引起的棉花轮纹斑病,是导致棉花早衰发生的主要成因之一,严重危害棉花生产。香豆素合成途径作为合成植保素的一条代谢支路,其产物能够抑制病原菌,增强植物的抗病性。为了揭示该途径在棉花抗链格孢菌中的作用,本文利用棉花皱缩病毒介导的基因沉默技术,通过对棉花香豆素合成途径中编码关键酶基因的沉默抑制,来解析其在棉花抗链格孢菌中的作用,所选择的基因有编码苯丙氨酸解氨酶(PAL)的基因Ghpal、肉桂-4-羟化酶(C4H)基因Ghc4h和4香豆酸-辅酶-A(4CL)的基因Gh4cl。结果显示,Ghpal、Ghc4h和Gh4cl基因受链格孢菌诱导转录,表明这些基因可能参与了棉花对链格孢菌的抗性反应。Ghpal、Ghc4h和Gh4cl基因分别被沉默后,结果显示各基因沉默植株对链格孢菌的抗性下降,表现为接种链格孢菌后,基因沉默植株的病情指数分别为43.36、38.27和44.78,显著高于野生型和VIGS空载体对照植株。这表明,Ghpal、Ghc4h和Gh4cl参与了棉花对链格孢菌的抗性反应。此外,离体条件下给Ghpal、Ghc4h和Gh4cl基因沉默植株叶片饲喂外源香豆素,结果发现基因沉默植株对链格孢菌的抗性得到恢复。这一结果进一步表明香豆素合成途径在棉花抗链格孢菌中起着重要作用。 相似文献
8.
草酸和BTH对黄瓜幼苗霜霉病抗性和胞间隙病程相关蛋白的诱导 总被引：7，自引：0，他引：7
本文研究了草酸和BTH(苯并噻二唑)溶液对黄瓜幼苗霜霉病的抗性诱导及病程相关蛋白的积累。结果表明:10mmol/L草酸或0.5mmol/LBTH均能诱导黄瓜对霜霉病产生局部和系统抗性,且抗性的持久期均在15d以上。BTH处理或接种霜霉菌都可系统诱导黄瓜叶片胞间隙液产生分子量分别为33、27和22kD的蛋白,而草酸没有诱导这3种蛋白的表达。BTH或草酸处理均可引起POD活性的升高,但和对照相比并没有新的POD同工酶表达,POD活性升高与黄瓜对霜霉病的抗性有关。 相似文献
9.
YASUHIRO YOGO 《Weed Biology and Management》2011,11(1):49-56
Lignin and its related metabolites play critical roles in plant growth and development. Thus, lignin biosynthesis has attracted interest as a novel target site of plant growth inhibitors. Chalcone has been shown to not only inhibit lignin biosynthesis in plants, but also to suppress the growth of many annual plant species. In order to know the direct effect of chalcone on plant metabolism, the effects of chalcone on the activities of key enzymes in lignin biosynthesis and on the related metabolites were clarified with a time‐course study by using light‐induced suspension cultures of soybean cells. The fresh weight and packed cell volume of the soybean cells were inhibited after 8 h of chalcone treatment. The activities of phenylalanine ammonia lyase (EC 4.3.1.24) and 4‐coumarate: coenzyme A ligase (4CL; EC 6.2.1.12) were largely inhibited 4 h after the treatment with 0.15 mmol L?1 chalcone. Unlike these two enzymes, the activity of cinnamyl alcohol dehydrogenase (EC 1.1.1.195) was not inhibited until 16 h after the chalcone treatment. The content of the 4CL substrates and lignin in the soybean cells became relatively lower than the control under the light condition within 4 h and 8 h after the chalcone treatment, respectively. These results suggest that the growth suppression of soybean cells is positively associated with the inhibition of lignin biosynthesis by exogenous chalcone. 相似文献
10.
绿色木霉菌T23对黄瓜枯萎病防治效果及其几种防御酶活性的影响 总被引：29，自引：0，他引：29
本文通过生物测定的方法初步研究了绿色木霉菌T23分生孢子和厚垣孢子对黄瓜枯萎病防治效果及黄瓜幼苗几种防御酶活性的影响。结果表明:黄瓜幼苗经木霉菌处理后,病情指数由33.69分别降至13.12和10.28,经木霉菌处理的黄瓜体内与抗性反应相关的苯丙氨酸解氨酶(PAL)、过氧化物酶(POD)、多酚氧化酶(PPO)及过氧化氢酶(CAT)活性有明显增加,其峰值分别为对照的2.75、2.49、2.42及15.84倍,说明生物防治过程中可能有木质素、植保素等抗性物质的参与。与分生孢子处理相比,木霉菌厚垣孢子处理的酶活高峰出现得晚,但酶活峰值高。 相似文献
11.
ABSTRACT The study was conducted to evaluate the potential of induced resistance to infestation of sunflower (Helianthus annuus L.) by the parasitic weed Orobanche cumana Wallr. Treatment of sunflower seeds with 40 ppm of benzo(1,2,3)thiadiazole-7-carbothioic acid S-methyl ester (BTH) for 36 h completely prevented infection in root chambers. In pot studies using 2.86 x 10(-4) g of Orobanche seeds per gram of soil as inoculum, the total number of O. cumana shoots was reduced by 84 and 95% in the 60-ppm BTH treatment in the first and second trial, respectively. Evaluation of the disease incidences revealed that attachment of O. cumana at the sunflower root and the stage of early penetration was reduced in the BTH-treated plants. Chemical analysis of root extracts revealed synthesis of the phytoalexin scopoletin and of hydrogen peroxide in the BTH-treated sunflower roots, but no increase in lignification. Western blot analysis demonstrated accumulation of the pathogenesis-related protein chitinase in roots and stems of induced resistant plants. These results show that the phenomenon of induced resistance is not restricted to viral, bacterial, and fungal disease and demonstrate the great potential of this protection strategy as an effective component of future plant production systems. 相似文献
12.
ABSTRACT The controversial role of silicon in plant disease resistance, described mostly as a passive mechanical protection, has been addressed. Conclusive evidence is presented that silicon is involved in the increased resistance of cucumber to powdery mildew by enhancing the antifungal activity of infected leaves. This antifungal activity was attributable to the presence of low-molecular-weight metabolites. One of these metabolites, described here as a phytoalexin, was identified as a flavonol aglycone rhamnetin (3,5,3',4'-tetrahydroxy-7-O-methoxyflavone). This is the first report of a phytoalexin for this chemical group in the plant kingdom and of a flavonol phytoalexin in cucumber, a chemical defense long believed to be nonexistent in the family Cucurbitaceae. The antifungal activity of leaf extracts was better expressed after acid hydrolysis, extending to another plant species the concept that some phytoalexins are synthesized as glycosylated phytoalexins or their precursors. 相似文献
13.
从不同处理方法、酶液pH变化等方面对果胶酶诱导黄瓜抗黑星病效能的影响进行了研究。结果表明，在果胶酶的4种不同处理试验中，以全株表面喷雾法处理的黄瓜黄化子叶黑星病的病情指数降低最多，浓度100和200U／ml的诱导防病效果分别为56．34％和64．13％；下胚轴注射、灌根和漫种等3种处理方法的诱导抗病效果均不显著。经果胶酶诱导处理的黄瓜绿苗叶片病情明显降低，而处理叶上部的未处理叶片发病情况与相应对照差别不大。该结果表明，果胶酶诱导黄瓜绿苗抗黑星病的作用属于局部诱导。果胶酶处理次数的增加可增强其诱导抗病效果，并延长诱导抗病时间。酶液pH变化对果胶酶的诱导抗病作用影响显著，pH5．5时，2个浓度的诱导效果最好，分别为51．87％和66．42％。 相似文献
14.
Acibenzolar-S-methyl (ASM) is a chemical activator of systemic disease resistance in plants. In this study, we used differential
display to identify ASM-inducible defense response genes involved in induced disease resistance. As a result, we cloned three
ASM-inducible genes from cucumber, encoding a chitinase, a putative protein disulfide isomerase and a putative mitochondrial-protein-like
protein. Expression of these genes was induced within 24 hr after treatment of cucumber leaves with ASM. These results suggest
that differential display is a useful tool for understanding the mode of action of ASM and defense responses.
Received 6 September 2000/ Accepted in revised form 11 April 2001 相似文献
15.
Cucumber Fusarium wilt (CFW), caused by the soil-borne fungus Fusarium oxysporum f. sp. cucumerium, is a serious disease in cucumber (Cucumis sativus) production worldwide. For the efficient control of the pathogenic fungi, a better understanding of its interaction and associated resistance mechanisms at the molecular level is required. Here, we report a comparative proteomics analysis of total root protein isolated from infected cucumber root of susceptible bulk (SB) and resistant bulk (RB) of cucumber generation F2. Two-dimensional gel electrophoresis (2-DE) coupled with MS/MS approaches identified 15 over-accumulated proteins from the RB plants. Identified proteins are mainly involved in defense and stress responses, oxidation reduction, metabolism and transport and other process. These proteins are likely to be a part of resistance-related protein network, playing different roles in cucumber disease resistance. Three vital clues regarding wilt resistance of C. sativus are gained from this study. First, jasmonic acid and redox signaling components were found in response to F. oxysporum infection in resistant plants. Second, the LRR family protein may play an important role in the defense reaction against CFW. Third, biotic and abiotic stress-related proteins were induced by the CFW fungus F. oxysporum, indicating the activation of common stress pathway. 相似文献
16.
草酸青霉菌果胶酶诱导黄瓜抗黑星病研究 总被引：6，自引：0，他引：6
本文就草酸青霉菌(Penicillium oxalicum)的固态发酵提取产物--果胶酶粗酶液诱导黄瓜对黑星病(Cladosporium cucumerinum)的抗性进行了研究。以果胶酶粗酶液喷雾处理4个感黑星病黄瓜品种的黄化苗,48 h后挑战接种黑星病菌孢子悬液,其中"中农5号"黄瓜品种表现的诱导抗病效果最好,诱抗效果达62.51%。不同浓度果胶酶诱导处理黄瓜,发现果胶酶浓度在20 U/mL时,可导致黄瓜发病略高于对照;在40~200 U/mL浓度范围内,诱导效果较为明显。通过研究果胶酶诱导抗病的时效性,表明诱导处理前接种病菌或诱导处理后0、6 h接种的各处理病情指数与对照间没有差别,而诱导处理12~72 h后接种病菌的,果胶酶的诱导抗病效果均很明显,诱抗效果达29.64%~60.02%。实验还表明,随着挑战接种压力的增大,果胶酶的诱导抗病效果降低。果胶酶不能抑制黑星病菌孢子萌发,相反可以促进孢子萌发和芽管生长。 相似文献
17.
Bacteria such as Pantoea agglomerans (Pa-AF2), Bacillus subtilis (Bs-271), Acinetobacter lwoffii (Al-113), and Pseudomonas fluorescens (Pf-CT2), originating from the vineyard, can induce defense responses and enhance resistance of grapevine against the fungal pathogen Botrytis cinerea. The perception of these bacteria by plant cells or tissues in relation to their activities remains unknown. In this study, we examined the relationships between the activity of each bacterium to induce or prime some defense responses, and its effectiveness to induce resistance in grapevine against B. cinerea. We showed that all selected bacteria are capable of inducing early oxidative burst and phytoalexin (trans-resveratrol and trans-ε-viniferin) production in grapevine cells and leaves. Pf-CT2 and Al-113 induced higher H(2)O(2) and trans-resveratrol accumulations, and were able to further prime plants for accelerated phytoalexin production after B. cinerea challenge. These two bacteria were also the most effective in inducing local and systemic resistance. A similar level of induced resistance was observed with live Pa-AF2 which also induced but not primed a greater accumulation of trans-resveratrol. However, Bs-271, which was less effective in inducing resistance, induced a lower trans-resveratrol synthesis, without priming activity. Treatment of grapevine cells with growing medium or crude extract of the bacteria quickly and strongly enhanced oxidative burst compared with the live bacteria. However, both treatments resulted in comparable amounts of phytoalexins and induced local and systemic resistance to B. cinerea as compared with those induced by living bacteria, with extracts from Pf-CT2 and Al-113 being the most effective. Together, these results indicate that induced resistance can be improved by treatment with bacteria or derived compounds which induced or primed plants for enhanced phytoalexin accumulation. 相似文献
18.
玫瑰黄链霉菌Strempomyces roseoflavus Men-myco-93-63是分离自马铃薯疮痂病自然衰退土壤中的一株拮抗链霉菌,该菌株及其代谢产物对多种重要的植物病原菌都具有较强的抑制作用,为了探明该生防菌产生的诱抗粗蛋白对黄瓜抗病性的诱导作用,采用离体叶片接种的方法,发现诱抗粗蛋白诱导黄瓜叶片灰霉病发病直径显著小于对照;经组织染色法和紫外分光光度计法测定,发现诱抗粗蛋白可以诱导黄瓜叶片中活性氧（ROS）的积累和超氧化物歧化酶（SOD）、过氧化物酶（POD）和多酚氧化酶（PPO）等抗病相关酶活性的显著提高;通过实时荧光定量PCR（qRT-PCR）测定,发现诱抗粗蛋白还可以诱导黄瓜叶片中PR-1a、PR-3、PR-9和NPR1等抗病相关基因表达的上调,试验结果表明Men-myco-93-63产生的诱抗粗蛋白能够诱导黄瓜抗病性的提升。 相似文献
19.
Lignin biosynthesis is essential for plant growth. 4‐Coumarate CoA ligase (4‐CL, EC6.2.1.12) is involved in the monolignol synthesis and occupies a key role in regulating carbon flow into the phenylpropanoid metabolism pathway. Naringenin, one of the metabolites in this pathway, is known as a potent in vitro inhibitor of 4‐CL. The growth of rice (Oryza sativa L. cv. Koshihikari), maize (Zea mays L. cv. Yellow corn) and Echinochloa oryzicola Vasing seedlings at the 2nd leaf stage was inhibited after continuous root application with 0.1 mmol L?1 naringenin for 1 week, although naringenin did not kill these gramineous plants. The highest inhibition of fresh weight increase was observed in maize, followed by rice and E. oryzicola. The symptoms in these plants were root browning, delay of leaf/root development and shoot dwarfing. Naringenin treatment increased the contents of 4‐CL substrates, cinnamic acid, 4‐coumaric acid, caffeic acid and ferulic acid from 1.2 to 7.2 times and from 1.2 to 3.5 times in shoots and roots, respectively, except for ferulic acid in E. oryzicola roots. It also caused a slight decrease of the lignin content and alteration of lignin constitutions in rice plants. These results suggested that the monolignol pathways after 4‐CL towards lignin has the possibility to be the novel action sites of plant growth retardants, although further investigations are needed to clarify the mode of action. 相似文献
20.
Kyutaro Kishimoto Yoko Nishizawa Yutaka Tabei Masami Nakajima Tadaaki Hibi Katsumi Akutsu 《Journal of General Plant Pathology》2004,70(6):314-320
A class III chitinase gene (CHI2) is induced in cucumber plants (Cucumis sativa L.) in response to infection by pathogenic microorganisms. Infection of Botrytis cinerea, causal agent of gray mold disease on cucumber, also induces CHI2 expression. To investigate whether CHI2 is involved in resistance to gray mold disease, transgenic cucumber plants were produced to overexpress the CHI2 gene. One line was analyzed in detail in terms of disease resistance. The transgenic cucumber plant (CC2) constitutively expressed CHI2 and reduced the symptoms of B. cinerea for 4 days after inoculation compared with nontransgenic plants. However, this inhibitory effect was not absolute, and CC2 eventually developed serious disease symptoms. Chitinase activity of the crude extract from CC2 leaves was higher than that from nontransgenic plants. A high-molecular-weight fraction containing CHI2 from CC2 leaves had fungistatic activity against B. cinerea. Interestingly, the low-molecular-weight fraction from CC2 leaves with CHI2 removed also had fungistatic activity against B. cinerea. Not only the introduced chitinase activity but also the endogenous defense reactions activated by overexpression of CHI2 may be involved in the enhanced gray mold disease resistance in CC2. 相似文献