Systemic Resistance In Rice Research Articles

Combination effects of tebuconazole with Bacillus subtilis to control rice false smut and the related synergistic mechanism.

Management of rice false smut is based mainly on chemical control, which poses many safety and environmental challenges. The other option, biological control with biofungicides, does not have such problems but is not as reliable because of low systemic ability, and lower and unstable efficacy. Therefore, it is necessary to combine application of chemical fungicides and biological control agents (BCAs) and elucidate their synergistic mechanism. A combination of tebuconazole and a proven BCA, Bacillus subtilis H158, was evaluated for control of rice false smut. Tebuconazole at low application rates stimulated growth of B. subtilis, prolonged the effective period of B. subtilis by enhancing its persistence on the surface of rice plants, accelerated biofilm formation of the BCA to facilitate colonization, promoted induced systemic resistance in rice by regulating defense-related enzymes and genes, and reduced the natural resistance of the pathogen by suppressing the key gene for fungal resistance to tebuconazole. However, at high application rates, tebuconazole had adverse effects on these factors and showed antagonistic combination effects with B. subtilis. The combination of B. subtilis with tebuconazole at low application rates showed great synergistic effects, but at high application rates showed only antagonistic effects in field experiments over two consecutive years. The combination of B. subtilis with tebuconazole had significant synergistic effects at low application rates. The synergistic effects are the result of multiple mechanisms involved in BCA, rice and the fungal pathogen. © 2022 Society of Chemical Industry.

Toxicity of Bacillus thuringiensis Strains Derived from the Novel Crystal Protein Cry31Aa with High Nematicidal Activity against Rice Parasitic Nematode Aphelenchoides besseyi.

The plant parasitic nematode, Aphelenchoides besseyi, is a serious pest causing severe damage to various crop plants and vegetables. The Bacillus thuringiensis (Bt) strains, GBAC46 and NMTD81, and the biological strain, FZB42, showed higher nematicidal activity against A. besseyi, by up to 88.80, 82.65, and 75.87%, respectively, in a 96-well plate experiment. We screened the whole genomes of the selected strains by protein-nucleic acid alignment. It was found that the Bt strain GBAC46 showed three novel crystal proteins, namely, Cry31Aa, Cry73Aa, and Cry40ORF, which likely provide for the safe control of nematodes. The Cry31Aa protein was composed of 802 amino acids with a molecular weight of 90.257 kDa and contained a conserved delta-endotoxin insecticidal domain. The Cry31Aa exhibited significant nematicidal activity against A. besseyi with a lethal concentration (LC50) value of 131.80 μg/mL. Furthermore, the results of in vitro experiments (i.e., rhodamine and propidium iodide (PI) experiments) revealed that the Cry31Aa protein was taken up by A. besseyi, which caused damage to the nematode’s intestinal cell membrane, indicating that the Cry31Aa produced a pore-formation toxin. In pot experiments, the selected strains GBAC46, NMTD81, and FZB42 significantly reduced the lesions on leaves by up to 33.56%, 45.66, and 30.34% and also enhanced physiological growth parameters such as root length (65.10, 50.65, and 55.60%), shoot length (68.10, 55.60, and 59.45%), and plant fresh weight (60.71, 56.45, and 55.65%), respectively. The number of nematodes obtained from the plants treated with the selected strains (i.e., GBAC46, NMTD81, and FZB42) and A. besseyi was significantly reduced, with 0.56, 0.83., 1.11, and 5.04 seedling mL−1 nematodes were achieved, respectively. Moreover, the qRT-PCR analysis showed that the defense-related genes were upregulated, and the activity of hydrogen peroxide (H2O2) increased while malondialdehyde (MDA) decreased in rice leaves compared to the control. Therefore, it was concluded that the Bt strains GBAC46 and NMTD81 can promote rice growth, induce high expression of rice defense-related genes, and activate systemic resistance in rice. More importantly, the application of the novel Cry31Aa protein has high potential for the efficient and safe prevention and green control of plant parasitic nematodes.

Dehydroascorbate induces plant resistance in rice against root-knot nematode Meloidogyne graminicola.

Ascorbic acid (AsA) is an important antioxidant in plants and regulates various physiological processes. In this study, we show that exogenous treatments with the oxidized form of AsA, that is, dehydroascorbate (DHA), activates induced systemic resistance in rice against the root‐knot nematode Meloidogyne graminicola, and investigate the molecular and biochemical mechanisms underlying this phenotype. Detailed transcriptome analysis on roots of rice plants showed an early and robust transcriptional response on foliar DHA treatment, with induction of several genes related to plant stress responses, immunity, antioxidant activity, and secondary metabolism already at 1 day after treatment. Quantitative and qualitative evaluation of H2O2 levels confirmed the appearance of a reactive oxygen species (ROS) burst on DHA treatment, both at the site of treatment and systemically. Experiments using chemical ROS inhibitors or scavengers confirmed that H2O2 accumulation contributes to DHA‐based induced resistance. Furthermore, hormone measurements in DHA‐treated plants showed a significant systemic accumulation of the defence hormone salicylic acid (SA). The role of the SA pathway in DHA‐based induced resistance was confirmed by nematode infection experiments using an SA‐signalling deficient WRKY45‐RNAi line and reverse transcription‐quantitative PCR on SA marker genes. Our results collectively reveal that DHA activates induced systemic resistance in rice against the root‐knot nematode M. graminicola, mediated through the production of ROS and activation of the SA pathway.

Assessment of Silicon- and Mycorrhizae- Mediated Constitutive and Induced Systemic Resistance in Rice, Oryza sativa L., against the Fall Armyworm, Spodoptera frugiperda Smith.

Induced resistance provides protection in plants against insect herbivores. Silicon and mycorrhizae often prime plant defenses and thereby enhance plant resistance against herbivores. In rice, Oryza sativa L., insect injury has been shown to induce resistance against future defoliators. However, it is unknown if silicon and mycorrhizae treatments in combination with insect injury result in greater induced resistance. Using the fall armyworm (FAW), Spodoptera frugiperda Smith, two experiments were conducted to investigate whether (1) silicon or mycorrhizae treatment alters resistance in rice and (2) induced systemic resistance in response to insect injury is augmented in silicon- or mycorrhizae- treated plants. In the first experiment, silicon treatment reduced FAW growth by 20% while mycorrhizae increased FAW growth by 8%. In the second experiment, insect injury induced systemic resistance, resulting in a 23% reduction in FAW larval weight gains on injured compared to uninjured plants, irrespective of treatment. Neither silicon nor mycorrhizae enhanced this systemic resistance in insect-injured plants. Furthermore, mycorrhizae resulted in the systemic increase of peroxidase (POD) and polyphenol oxidase (PPO) activities, and injury caused a slight decrease in these enzyme activities in mycorrhizae plants. Silicon treatment did not result in a stronger induction of POD and PPO activity in injured plants. Taken together, these results indicate a lack of silicon and mycorrhizae priming of plant defenses in rice. Regardless of injury, silicon reduced FAW weight gains by 36%. Based on these results, it appears silicon-mediated biomechanical rather than biochemical defenses may play a greater role in increased resistance against FAW in rice.

Induced Resistance by Ascorbate Oxidation Involves Potentiating of the Phenylpropanoid Pathway and Improved Rice Tolerance to Parasitic Nematodes.

Anticipating an increased ecological awareness, scientists have been exploring new strategies to reduce the use of chemical pesticides to control pests and diseases. Triggering the intrinsic plant defense system is one of the promising strategies to reduce yield loss by pathogenic organisms, such as nematodes. Ascorbate oxidase (AO) enzyme plays an important role in plant defense by regulating the apoplastic ascorbate/dehydroascorbate (DHA) ratio via the ascorbate oxidation process. Ascorbate oxidation is known to induce systemic resistance in rice against parasitic root-knot nematodes (RKN). Here, we sought to evaluate if AO- or DHA-induced resistance (IR) against RKN M. graminicola involves activation of the phenylpropanoid pathway and whether this IR phenotype has potential effects on growth of rice seedlings under stressed and unstressed conditions. Our results show that AO/DHA-IR against these parasitic nematodes is dependent on activation of phenylalanine ammonia lyase (PAL). However, application of reduced ascorbic acid (AA) did not induce this response. Gene expression analysis via qRT-PCR showed that OsPAL2 and OsPAL4 are highly expressed in AO/DHA-sprayed nematode-infected roots and PAL-activity measurements confirmed that AO/DHA spraying triggers the plants for primed activation of this enzyme upon nematode infection. AO/DHA-IR is not effective in plants sprayed with a chemical PAL inhibitor confirming that AO/DHA-induced resistance is dependent on PAL activity. Improved plant growth and low nematode infection in AO/DHA-sprayed plants was found to be correlated with an increase in shoot chlorophyll fluorescence (Fv/Fm), chlorophyll index (ChlIdx), and modified anthocyanin reflection index which were proven to be good above-ground parameters for nematode infestation. A detailed growth analysis confirmed the improved growth of AO/DHA-treated plants under nematode-infected conditions. Taken together, our results indicate that ascorbate oxidation enhances the phenylpropanoid-based response to nematode infection and leads to a tolerance phenotype in treated rice plants.

The cyclic lipopeptide orfamide induces systemic resistance in rice to Cochliobolus miyabeanus but not to Magnaporthe oryzae.

The Pseudomonas- derived cyclic lipopeptide orfamide can induce resistance to Cochliobolus miyabeanus but not to Magnaporthe oryzae in rice. Abscisic acid signaling is involved in the induced systemic resistance response triggered by orfamide. Diverse natural products produced by beneficial Pseudomonas species have the potential to trigger induced systemic resistance (ISR) in plants, and thus may contribute to control of diseases in crops. Some beneficial Pseudomonas spp. can produce cyclic lipopeptides (CLPs), amphiphilic molecules composed of a fatty acid tail linked to an oligopeptide which is cyclized. CLPs can have versatile biological functions, but the capacity of Pseudomonas-derived CLPs in triggering ISR responses has barely been studied. Pseudomonas protegens and related species can produce orfamide-type CLPs. Here we show that in rice, orfamides can act as ISR elicitors against the necrotrophic fungus Cochliobolus miyabeanus, the causal agent of brown spot disease, but are not active against the blast fungus Magnaporthe oryzae. Orfamide A can trigger early defensive events and activate transcripts of defense-related genes in rice cell suspension cultures, but does not cause cell death. Further testing in rice cell suspension cultures and rice plants showed that abscisic acid signaling, the transcriptional activator OsWRKY4 and pathogenesis-related protein PR1b are triggered by orfamide A and may play a role in the ISR response against C. miyabeanus.

Exogenous proteinogenic amino acids induce systemic resistance in rice.

BackgroundPlant immune responses can be induced by endogenous and exogenous signaling molecules. Recently, amino acids and their metabolites have been reported to affect the plant immune system. However, how amino acids act in plant defense responses has yet to be clarified. Here, we report that treatment of rice roots with amino acids such as glutamate (Glu) induced systemic disease resistance against rice blast in leaves.ResultsTreatment of roots with Glu activated the transcription of a large variety of defense-related genes both in roots and leaves. In leaves, salicylic acid (SA)-responsive genes, rather than jasmonic acid (JA) or ethylene (ET)-responsive genes, were induced by this treatment. The Glu-induced blast resistance was partially impaired in rice plants deficient in SA signaling such as NahG plants expressing an SA hydroxylase, WRKY45-knockdown, and OsNPR1-knockdown plants. The JA-deficient mutant cpm2 exhibited full Glu-induced blast resistance.ConclusionsOur results indicate that the amino acid-induced blast resistance partly depends on the SA pathway but an unknown SA-independent signaling pathway is also involved.Electronic supplementary materialThe online version of this article (doi:10.1186/s12870-016-0748-x) contains supplementary material, which is available to authorized users.

Induction of systemic resistance in rice by leaf extracts of Datura metel against sheath blight disease

The leaf extracts of Datura metel [both aqueous leaf extract (ALE) and ethanolic leaf extract (ELE)] were observed here to find if they can induce systemic resistance in the rice commonly found in Eastern India. The results showed that after the treatment, the enzyme activities of all the defence-related enzymes increased to a certain level even without pathogenic infection in comparison with non-treated seedlings and then, maintain at constant level throughout the study period. When treated seedlings were infected with Rhizoctonia solani, the enzyme activities were increased more than in uninfected seedlings. The elevated enzyme activities gave the indication of an induced systemic resistance in rice. The ELE of D. metel showed better induction effect than ALE.

Plant growth-promoting rhizobacteria mediate induced systemic resistance in rice against bacterial leaf blight caused by Xanthomonas oryzae pv. oryzae

Seven Bacillus plant growth-promoting rhizobacteria spp. were evaluated for growth promotion and induced systemic resistance in rice against Xanthomonas oryzae pv. oryzae (Xoo). The identities of colonies of X. oryzae pv. oryzae grown on mXOS and PSA medium were confirmed by PCR employing specific primers TXTF and TXT4R. Among the seven strains tested as fresh suspensions, talc and sodium alginate formulations under laboratory and green house conditions, maximum germination of 86% was recorded after seed treatments with fresh suspension of Bacillus subtilis GBO3 followed by 85% germination treated with Bacillus pumilus SE34 in comparison to only 71% germination in the untreated controls. Similarly, the maximum vigor index of 1374 was obtained by seed treatment with fresh suspensions of B. subtilis strain GBO3 followed by treatments with strain SE34 with vigor index of 1323 in contrast to an index of only 834 observed in untreated controls. Among the treatments, seed treatments with fresh suspension of seven strains resulted in better germination and vigor assessments than talc based or sodium alginate formulations. Seed treatments with fresh suspension of strain SE34 gave 71% protection, followed by B. subtilis GBO3 and B. pumilus T4 with 58% and 52% protection, respectively, compared to the untreated controls. Seed treatments with talc based formulation of SE34 gave 66% protection, while GBO3 and T4 resulted in 52% and 50% protection, respectively, with similar formulation. Seed treatment with talc and sodium alginate formulations of strain SE34 gave 58% protection followed by GBO3 with 40% protection. Seed treatment with fresh suspensions of strains SE34 and GBO3 followed by challenge inoculations with Xoo increased accumulation of phenylalanine ammonia lyase, peroxidase and polyphenol oxidase compared to untreated control seedlings. Thus, the results of the present study suggest that the PGPR strains used as fresh suspensions and powdered formulations may have commercial potential in plant growth promotion and in management of rice bacterial leaf blight disease.

Induction of systemic resistance in rice to bacterial blight by 1,2,3-benzothiadiazole 7-carbothioic acid-S-methyl ester (BTH) treatments

Use of BTH to evaluate the disease severity and induction of systemic resistance in rice to bacterial blight caused by Xanthomonas oryzae pv. oryzae is investigated. A new batch of 25 isolates of Xanthomonas oryzae pv. oryzae was obtained from infected rice lead tissues collected from Pattambi, Kerala, south India. Their identification was confirmed by the plant inoculation test on to IR24 rice plants which produced characteristic bacterial blight lesions. Among the 25 of X.o. pv. oryzae, four of the isolates were also virulent to IRBB21 rice plants (a near isogenic line of IR24) which carry the Xa-21 gene for BB resistance. The results confirm that there are pathogen strains in India which can overcome Xa-21. Development of BB lesions developed in IR24 (BB susceptible) plants after they were treated with BTH applications either as seed treatment or as foliar spray at 0.1, 0.5, 0.1 and 2.0 mM concentrations showed that even at 2.0 mM concentrations, IR24 plants were still susceptible to the pathogen. There was very little or marginal effect of BTH on the induction of resistance to BB in IR24 rice plants. When the same concentrations of BTH were applied to IRBB21 (Xa-21) rice plants, they showed pronounced triggering of systemic resistance to BB pathogen even at 0.1 mM concentration of BTH applied either as seed treatment or as foliar spry. Disease severity index was reduced to 5 (against a score of 9 in untreated) and there was 85–86% reduction in BB incidence in plants that received 0.1 mM BTH. These results provide evidence that BTH-induced systemic resistance complements the R-gene resistance in IRBB21 plants but not in IR24 rice plants.

Pseudomonas fluorescens WCS374r-Induced Systemic Resistance in Rice against Magnaporthe oryzae Is Based on Pseudobactin-Mediated Priming for a Salicylic Acid-Repressible Multifaceted Defense Response

Selected strains of nonpathogenic rhizobacteria can reduce disease in foliar tissues through the induction of a defense state known as induced systemic resistance (ISR). Compared with the large body of information on ISR in dicotyledonous plants, little is known about the mechanisms underlying rhizobacteria-induced resistance in cereal crops. Here, we demonstrate the ability of Pseudomonas fluorescens WCS374r to trigger ISR in rice (Oryza sativa) against the leaf blast pathogen Magnaporthe oryzae. Using salicylic acid (SA)-nonaccumulating NahG rice, an ethylene-insensitive OsEIN2 antisense line, and the jasmonate-deficient mutant hebiba, we show that this WCS374r-induced resistance is regulated by an SA-independent but jasmonic acid/ethylene-modulated signal transduction pathway. Bacterial mutant analysis uncovered a pseudobactin-type siderophore as the crucial determinant responsible for ISR elicitation. Root application of WCS374r-derived pseudobactin (Psb374) primed naive leaves for accelerated expression of a pronounced multifaceted defense response, consisting of rapid recruitment of phenolic compounds at sites of pathogen entry, concerted expression of a diverse set of structural defenses, and a timely hyperinduction of hydrogen peroxide formation putatively driving cell wall fortification. Exogenous SA application alleviated this Psb374-modulated defense priming, while Psb374 pretreatment antagonized infection-induced transcription of SA-responsive PR genes, suggesting that the Psb374- and SA-modulated signaling pathways are mutually antagonistic. Interestingly, in sharp contrast to WCS374r-mediated ISR, chemical induction of blast resistance by the SA analog benzothiadiazole was independent of jasmonic acid/ethylene signaling and involved the potentiation of SA-responsive gene expression. Together, these results offer novel insights into the signaling circuitry governing induced resistance against M. oryzae and suggest that rice is endowed with multiple blast-effective resistance pathways.

Pseudomonas aeruginosa inducing rice resistance againstRhizoctonia solani: Production of salicylic acid and peroxidases

Three isolates of Pseudomonas aeruginosa were used for seed treatment of rice; all showed plant growth promoting activity and induced systemic resistance in rice against Rhizoctonia solani G5 and increased seed yield. Production of salicylic acid (Sal) by P. aeruginosa both in vitro and in vivo was quantified with high performance liquid chromatography. All three isolates produced more Sal in King's B broth than in induced roots. Using a split root system, more Sal accumulated in root tissues of bacterized site than in distant roots on the opposite site of the root system after 1 d, but this difference decreased after 3 d. Sal concentration 0-200 g/L showed no inhibition of mycelial growth of R. solani in vitro, while at > or =300 g/L it inhibited it. P. aeruginosa-pretreated rice plants challenged inoculation with R. solani (as pathogen), an additional increase in the accumulation of peroxidase was observed. Three pathogenesis-related peroxidases in induced rice plants were detected; molar mass of these purified peroxidases was 28, 36 and 47 kDa. Purified peroxidase showed antifungal activity against phytopathogenic fungi R. solani, Pyricularia oryzae and Helminthosporium oryzae.

Antimicrobial activity and induction of systemic resistance in rice by leaf extract of Datura metel against Rhizoctonia solani and Xanthomonas oryzae pv. oryzae

The leaf extracts of Datura metel significantly reduced the in vitro growth of Rhizoctonia solani (RS7, Anastomosis group AG1) and Xanthomonas oryzae pv. o ryzae ( Xoo). Methanol extract exhibited the best control of the pathogens recording 10–35% more toxicity than aqueous extract. Foliar application of leaf extracts effectively reduced the incidence of sheath blight and bacterial blight diseases of rice under greenhouse condition. Pre-inoculation applications of leaf extract were found better than post-inoculation applications. Induction of systemic resistance as evident from the increased accumulation of pathogenesis related (PR) proteins and other defense related compounds was observed in rice plants following application of leaf extract of D. metel and challenge inoculation with either R. solani or Xoo. The methanol extract of D. metel when subjected to thin layer chromatography (TLC) showed a spot with relative front (Rf) value of 0.705. Well pronounced antibacterial activity against Xoo was demonstrated by the compound eluted from the region corresponding to this spot. The molecular ion peak and the fragmentation peaks of the eluted compound analyzed by mass spectrometry correspond to the already published mass spectrum of daturilin (Siddiqui et al., Phytochemistry 26: 2641-2643, 1987), a withanolide compound from D. metel.

Induction of systemic resistance in rice against sheath blight disease by Pseudomonas fluorescens

Two Pseudomonas fluorescens strains viz., PF1 and FP7 which inhibited the mycelial growth of sheath blight fungus Rhizoctonia solani and increased the seedling vigour of rice plants in vitro were selected for assessing induced systemic resistance (ISR) against R. solani in rice. The Pseudomonas application as a bacterial suspension or a talc-based formulation through seed, root, soil and foliar application either alone or in combination (seed+root+soil+foliar) effectively reduced sheath blight disease incidence, promoted plant growth and ultimately increased yields under glasshouse or field conditions. Efficacy of Pseudomonas strains against R. solani was comparable to that of the fungicide carbendazim, which is normally used in the field to manage the disease. Pseudomonas treatment of rice cv IR50 led to induction of systemic resistance against R. solani, as a result of increase in chitinase and peroxidase activity. However, the extent of increase varied between treatments, Pseudomonas strains used and their duration. Though two chitinase isoforms (35 and 28 kDa) and five peroxidase isozymes (PO1–PO5) were found to be associated with the ISR, 35 kDa chitinase and three peroxidase isozymes (PO3–PO5) were established as the major determinants of ISR. Although a single application of a Pseudomonas strain resulted in ISR, the combined application through all of the four (seed, root, soil and foliar) methods increased the durability of ISR in rice plants. In addition, the Pseudomonas strains produced chitinase in the culture medium. It is presumed that the induced chitinase, peroxidase and bacterial chitinase may be either directly or indirectly involved in the reduction of sheath blight disease development in rice.