Jasmonic acid and salicylic acid activate a common defense system in rice

Jasmonic acid (JA) and salicylic acid (SA) play important roles in plant defense systems. JA and SA signaling pathways interact antagonistically in dicotyledonous plants, but, the status of crosstalk between JA and SA signaling is unknown in monocots. Our rice microarray analysis showed that more than half of the genes upregulated by the SA analog BTH are also upregulated by JA, suggesting that a major portion of the SA-upregulated genes are regulated by JA-dependent signaling in rice. A common defense system that is activated by both JA and SA is thus proposed which plays an important role in pathogen defense responses in rice.     

Herbivore-induced rice resistance against rice blast mediated by salicylic acid

In agro-ecosystems,plants are important mediators of interactions between their associated herbivorous insects and microbes,and any change in plants induced by one species may lead to cascading effects on interactions with other species.Often,such effects are regulated by phytohormones such as jasmonic acid(JA)and salicylic acid(SA).Here,we investigated the tripartite interactions among rice plants,three insect herbivores(Chilo suppressalis,Cnaphalocrocis medinalis or Nilapai-vata lugens),and the causal agent of rice blast disease,the fungus Magnaporthe oryzae.We found that pre-infestation of rice by C.suppressalis or N.lugens but not by C.medinalis conferred resistance to M.oryzae.For C.suppressalis and N.lugens,insect infestation without fungal inoculation induced the accumulation of both JA and SA in rice leaves.In contrast,infestation by C.medinalis increased JA levels but reduced SA levels.The exogenous application of SA but not of JA conferred resistance against M.oryzae.These results suggest that preinfestation by C suppressalis or N.lugens conferred resistance against M.oryzae by increasing SA accumulation.These findings enhance our understanding of the interactions among rice plant,insects and pathogens,and provide valuable information for developing an ecologically sound strategy for controlling rice blast.     

CRISPR/Cas9-mediated simultaneous mutation of three salicylic acid 5-hydroxylase (OsS5H) genes confers broad-spectrum disease resistance in rice

Salicylic acid (SA) is an essential plant hormone that plays critical roles in basal defence and amplification of local immune responses and establishes resistance against various pathogens. However, the comprehensive knowledge of the salicylic acid 5-hydroxylase (S5H) in rice-pathogen interaction is still elusive. Here, we reported that three OsS5H homologues displayed salicylic acid 5-hydroxylase activity, converting SA into 2,5-dihydroxybenzoic acid (2,5-DHBA). OsS5H1, OsS5H2, and OsS5H3 were preferentially expressed in rice leaves at heading stage and responded quickly to exogenous SA treatment. We found that bacterial pathogen Xanthomonas oryzae pv. oryzae (Xoo) strongly induced the expression of OsS5H1, OsS5H2, and OsS5H3. Rice plants overexpressing OsS5H1, OsS5H2, and OsS5H3 showed significantly decreased SA contents and increased 2,5-DHBA levels, and were more susceptible to bacterial blight and rice blast. A simple single guide RNA (sgRNA) was designed to create oss5h1oss5h2oss5h3 triple mutants through CRISPR/Cas9-mediated gene mutagenesis. The oss5h1oss5h2oss5h3 exhibited stronger resistance to Xoo than single oss5h mutants. And oss5h1oss5h2oss5h3 plants displayed enhanced rice blast resistance. The conferred pathogen resistance in oss5h1oss5h2oss5h3 was attributed to the significantly upregulation of OsWRKY45 and pathogenesis-related (PR) genes. Besides, flg22-induced reactive oxygen species (ROS) burst was enhanced in oss5h1oss5h2oss5h3. Collectively, our study provides a fast and effective approach to generate rice varieties with broad-spectrum disease resistance through OsS5H gene editing.     

Chemical induction of disease resistance in rice is correlated with the expression of a gene encoding a nucleotide binding site and leucine-rich repeats

Probenazole (3-allyloxy-1,2-benzisothiazole-1,1-dioxide) is an agricultural chemical primarily used to prevent rice blast disease. Probenazole-treated rice acquires resistance to blast fungus irrespective of the rice variety. The chemical is applied prophylactically, and is thought to induce or bolster endogenous plant defenses. However, the mechanisms underlying this effect have not been established. To understand the mode of the chemical's action, we screened for novel probenazole-responsive genes in rice by means of differential display and identified a candidate gene, RPR1. RPR1 contains a nucleotide binding site and leucine-rich repeats, thus sharing structural similarity with known disease resistance genes. The expression of RPR1 in rice can be up-regulated by treatment with chemical inducers of systemic acquired resistance (SAR) and by inoculation with pathogens. RPR1-related sequences in rice varieties seem to be varied in sequence and/or expression, indicating that RPR1 itself is not a crucial factor for induced resistance in rice. However, Southern blot analysis revealed the existence of homologous sequences in all varieties examined. While the role of RPR1 has yet to be clarified, this is the first report of the identification of a member of this gene class and its induction during the systemic expression of induced disease resistance.     

Integrated control of rice kernel smut disease using plant extracts and salicylic acid

The effects of salicylic acid (SA) at three concentrations i.e. 2.5, 5 and 7 mM and plant extracts from pick tooth (Ammi visnaga), liquorice (Glycyrrhiza glabra), artemisia (Artemisia judaica), mint (Mentha viridis), clove (Syzygium aromaticum) and blue gum (Eucalyptus globulus) on the infection of rice kernel smut disease caused by Tilletia barclayana were studied. Spraying of rice plants with different concentrations of SA at seven days before infection was the most effective treatment against pathogen infection. Among all plant extract treatments, M. viridis and S. aromaticum were the most effective treatments. Additionally, our results showed increased levels of peroxidase, polyphenol oxidase, phenylalanine ammonia lyase and chitinase as well as total protein contents in the treated plants compared with the control. In conclusion, accumulations of these oxidative enzymes in plants treated with SA and plant extracts provide their role in the activation of induced resistance against T. barclayana.     

Integration of Pseudomonas fluorescens and salicylic acid improves citrus canker disease management caused by Xanthomonas citri subsp citri-A*

The individual and combined effects of Pseudomonas fluorescens (Pf) and salicylic acid (SA) were investigated for control of citrus bacterial canker (CBC). Both treated plants with copper hydroxide and untreated ones were used as controls. Mexican lime (Citrus aurantifolia) seedlings were treated with SA at 10 mM, Pf and distilled water. Plants were initially inoculated with Xanthomonas citri subsp citri 72 h post treatments. Results indicated that the Pf and SA treatment controlled CBC more effectively compared to separately applying Pf or SA. The application of Pf in combination with SA significantly reduced lesion number per leaf (72%) and disease severity (84%). Significant changes in the activities of peroxidase and catalase were found. In conclusion, the integration of Pf with SA complements each other and can be applied to manage citrus canker disease in conjunction with other control programmes.     

NbALD1 mediates resistance to turnip mosaic virus by regulating the accumulation of salicylic acid and the ethylene pathway in Nicotiana benthamiana

AGD2-LIKE DEFENCE RESPONSE PROTEIN 1 (ALD1) triggers plant defence against bacterial and fungal pathogens by regulating the salicylic acid (SA) pathway and an unknown SA-independent pathway. We now show that Nicotiana benthamiana ALD1 is involved in defence against a virus and that the ethylene pathway also participates in ALD1-mediated resistance. NbALD1 was up-regulated in plants infected with turnip mosaic virus (TuMV). Silencing of NbALD1 facilitated TuMV infection, while overexpression of NbALD1 or exogenous application of pipecolic acid (Pip), the downstream product of ALD1, enhanced resistance to TuMV. The SA content was lower in NbALD1-silenced plants and higher where NbALD1 was overexpressed or following Pip treatments. SA mediated resistance to TuMV and was required for NbALD1-mediated resistance. However, on NahG plants (in which SA cannot accumulate), Pip treatment still alleviated susceptibility to TuMV, further demonstrating the presence of an SA-independent resistance pathway. The ethylene precursor, 1-aminocyclopropanecarboxylic acid (ACC), accumulated in NbALD1-silenced plants but was reduced in plants overexpressing NbALD1 or treated with Pip. Silencing of ACS1, a key gene in the ethylene pathway, alleviated the susceptibility of NbALD1-silenced plants to TuMV, while exogenous application of ACC compromised the resistance of Pip-treated or NbALD1 transgenic plants. The results indicate that NbALD1 mediates resistance to TuMV by positively regulating the resistant SA pathway and negatively regulating the susceptible ethylene pathway.     

Overexpression of AtSGT1, an Arabidopsis salicylic acid glucosyltransferase, leads to increased susceptibility to Pseudomonas syringae

We reported previously that a recombinant salicylic acid (SA) glucosyltransferase1 (AtSGT1) from Arabidopsis thaliana catalyzes the formation of both SA 2-O-beta-D-glucoside (SAG) and the glucose ester of SA (SGE). Here, transgenic Arabidopsis plants overexpressing AtSGT1 have been constructed, and their phenotypes analyzed. Compared to wild-type plants, transgenic plants showed an increased susceptibility to Pseudomonas syringae and reduced the accumulation levels of both free SA and its glucosylated forms (SAG and SGE). On the other hand, the overexpression increased the levels of methyl salicylate (MeSA) and methyl salicylate 2-O-beta-D-glucoside (MeSAG), and also induced SA carboxyl methyltransferase1 (AtBSMT1) expression, whose products catalyze the conversion of SA to MeSA. Our data indicate that reduced resistance by AtSGT1 overexpression results from a reduction in SA content, which is at least in part caused by increases in MeSAG and MeSA levels at the expense of SA. Our study also suggests that genetic manipulation of AtSGT1 can be utilized as an important regulatory tool for pathogen control.     

Abscisic acid modulates salicylic acid biosynthesis for systemic acquired resistance in tomato

Among the regulatory mechanisms of systemic acquired resistance (SAR) in tomato, antagonistic interaction between salicylic acid (SA) and abscisic acid (ABA) signaling pathways was investigated. Treatment with 1,2-benzisothiazol-3(2H)-one1,1-dioxide (BIT) induced SAR in tomato thorough SA biosynthesis. Pretreatment of ABA suppressed BIT-induced SAR including SA accumulation, suggesting that ABA suppressed SAR by inhibiting SA biosynthesis.     

Endogenous salicylic acid protects rice plants from oxidative damage caused by aging as well as biotic and abiotic stress

Salicylic acid (SA) is a key endogenous signal that mediates defense gene expression and disease resistance in many dicotyledonous species. In contrast to tobacco and Arabidopsis, which contain low basal levels of SA, rice has two orders of magnitude higher levels of SA and appears to be insensitive to exogenous SA treatment. To determine the role of SA in rice plants, we have generated SA-deficient transgenic rice by expressing the bacterial salicylate hydroxylase that degrades SA. Depletion of high levels of endogenous SA in transgenic rice does not measurably affect defense gene expression, but reduces the plant's capacity to detoxify reactive oxygen intermediates (ROI). SA-deficient transgenic rice contains elevated levels of superoxide and H2O2, and exhibits spontaneous lesion formation in an age- and light-dependent manner. Exogenous application of SA analog benzothiadiazole complements SA deficiency and suppresses ROI levels and lesion formation. Although an increase of conjugated catechol was detected in SA-deficient rice, catechol does not appear to significantly affect ROI levels based on the endogenous catechol data and exogenous catechol treatment. When infected with the blast fungus (Magnaporthe grisea), SA-deficient rice exhibits increased susceptibility to oxidative bursts elicited by avirulent isolates. Furthermore, SA-deficient rice is hyperresponsive to oxidative damage caused by paraquat treatment. Taken together, our results strongly suggest that SA plays an important role to modulate redox balance and protect rice plants from oxidative stress.     

Tenuazonic acid,toxin of rice blast fungus,induces disease resistance and reactive oxygen production in plants

Effects of tenuazonic acid (TA) on rice leaf segments and on their interaction with compatible races of the blast fungus (Magnaporthe grisea, former name is Pyricularia oryzae) were studied. TA induced small brown necrotic spots on leaves Application of TA (1 or 5 mM) to leaves in mixtures with M. grisea spores induced a local disease resistance, which reduced the frequency of compatible lesions. TA was not fungitoxic but, in contact with the leaf, increased the capability of leaf diffusates to inhibit germination of M. grisea spores. In the infected leaves, the diffusate fungitoxicity was higher than in the healthy ones. Antioxidant enzymes, superoxide dismutase and catalase, and scavengers of hydroxyl radical, mannitol and formate, strongly inhibited the TA-induced diffusate fungitoxicity. It is suggested that the disease resistance induced by TA is mediated, at least partially, by generation of reactive oxygen species by rice leaves, which inhibit the development of the fungus directly or indirectly.     

Proposed definitions related to induced disease resistance

A clear definition of induced disease resistance is lacking, even though the area has been extensively researched and described. The lack of a precise definition leads to potential confusion on whether an underlying mechanism for biological control is induced resistance. We propose definitions, which were developed at a NATO Advanced Research Workshop on biological control, for induced disease resistance and related terms. These definitions are intended to invoke debate and increase effective communication among investigators of induced resistance.     

茉莉酸及其甲酯在植物诱导抗病性中的作用

茉莉酸类物质被认为是植物抗病防卫反应的内源及中间信号分子。本文介绍了茉莉酸及其甲酯在植物抗病性中的作用,从它们在体内激活的代谢途径及相关基因表达探讨有关作用机制以及有可能在农业上应用的前景。     

Imprimatins A and B: Novel plant activators targeting salicylic acid metabolism in Arabidopsis thaliana

Plant activators are agrochemicals that protect plants from a broad range of pathogens by activating the plant immune system. Unlike pesticides, they do not target pathogens; therefore, plant activators provide durable effects that are not overcome by pathogenic microbes. Although certain plant activators have been applied to paddy fields for more than 30 years, the molecular basis of the underlying immune induction are unclear. From the screening of 10,000 diverse chemicals by a high-throughput screening procedure to identify compounds that specifically enhance pathogen-induced cell death in Arabidopsis cultured cells, we identified 7 compounds, which we designated as immune priming chemicals (Imprimatins). These compounds increased disease resistance against pathogenic Pseudomonas bacteria in Arabidopsis plants. Pretreatments increased the accumulation of endogenous salicylic acid (SA) but reduced its metabolite, SA-O-β-D-glucoside (SAG). Imprimatins inhibited the enzymatic activities of 2 SA glucosyltransferases (SAGTs) in vitro at concentrations effective for immune priming. Single and double knockout Arabidopsis plants for both SAGTs consistently exhibited enhanced disease resistance and SA accumulation. Our results demonstrate that the control of the free SA pool through SA-inactivating enzymes can be a useful methodology to confer disease resistance in plants. SAGTs can pave the way for target-based discovery of novel crop protectants.     

外源水杨酸和茉莉酸诱导巨峰葡萄抗根瘤蚜

【目的】水杨酸和茉莉酸在植物诱导防御虫害反应中发挥着重要作用。本研究旨在探讨水杨酸和茉莉酸诱导葡萄对根瘤蚜的抗性。【方法】以盆栽巨峰为试材,在接种葡萄根瘤蚜Daktulosphaira vitifoliae的同时喷施水杨酸和茉莉酸,测定和评估了对根瘤蚜生长发育及产卵量的影响,以及对葡萄根系抗氧化相关酶［过氧化物酶(POD)和过氧化氢酶(CAT）］活性、丙二醛（MDA）含量、新梢生长量及光合速率的影响。【结果】水杨酸和茉莉酸诱导处理显著降低了根瘤蚜卵量及下代1,2龄若蚜总数,接种35 d后根瘤蚜的产卵量分别减少了41.35％和50.00％,1龄和2龄若蚜总数分别减少了42.31％和50.00％;根瘤蚜侵染后根系中POD和CAT活性均呈先升高后降低的趋势,且水杨酸和茉莉酸诱导处理在各测定时期均高于仅接种根瘤蚜处理;水杨酸和茉莉酸处理的根系中MDA含量在各测定时期均低于仅接种根瘤蚜处理;水杨酸和茉莉酸处理降低了根瘤蚜侵染对植株地上部生长及光合的抑制。接种处理后第30天,仅接种根瘤蚜处理的植株地上部生长量减少了48.11%,光合速率降低了58.77％,而水杨酸和茉莉酸处理后的新梢生长量分别减少了31.57％和25.71％,光合速率分别降低了32.89％和24.67％。【结论】叶片喷洒茉莉酸和水杨酸能够降低根瘤蚜种群密度,并提高葡萄根系活性氧清除能力和防御酶活性,缓解树势衰退。     

Probenazole-induced accumulation of salicylic acid confers resistance to Magnaporthe grisea in adult rice plants

Probenazole (PBZ) is the active ingredient of Oryzemate, an agrochemical which is used for the protection of rice plants from Magnaporthe grisea (blast fungus). While PBZ was reported to function upstream of salicylic acid (SA) in Arabidopsis, little is known about the mechanism of PBZ-induced resistance in rice. The role of SA in blast fungus resistance is also unclear. The recommended application period for Oryzemate is just before the Japanese rainy season, at which time rice plants in the field have reached the 8-leaf stage with adult traits. Thus, the involvement of SA in PBZ-induced resistance was studied in compatible and incompatible blast fungus-rice interactions at two developmentally different leaf morphology stages. Pre-treatment of inoculated fourth leaves of young wild-type rice plants at the 4-leaf stage with PBZ did not influence the development of whitish expanding lesions (ELs) in the susceptible interaction without the accumulation of SA and pathogenesis-related (PR) proteins. However, PBZ pre-treatment increased accumulation of SA and PR proteins in the eighth leaves of adult plants at the 8-leaf stage, resulting in the formation of hypersensitive reaction (HR) lesions (HRLs). Exogenous SA induced resistance in adult but not young plants. SA concentrations in blast fungus-inoculated young leaves were essentially the same in compatible and incompatible interactions, suggesting that PBZ-induced resistance in rice is age-dependently regulated via SA accumulation.