Role of SABP2 in Systemic Acquired Resistance Induced by Acibenzolar-S-Methyl in Plants.

Plants have evolved an efficient mechanism to defend themselves against pathogens. Many biotic and abiotic agents have been shown to induce defense mechanism in plants. Acibenzolar-S-Methyl (ASM) is a commercially available chemical inducer of local and systemic resistance (SAR) response in plants. ASM functioning at molecular level is mostly unclear. This research was designed to investigate the mechanism of ASM action in plants. It was hypothesized that SABP2, a plant protein, plays an important role in ASM-mediated defense signaling. Biochemical studies were performed to test the interaction between SABP2 and ASM. Transgenic SABP2-silenced tobacco plants were used to determine the role of SABP2 in SAR induced by ASM. The expression of PR-1 proteins was used as a marker for SAR induction. Results showed that SABP2 converts ASM into acibenzolar that induces the expression of PR-1 proteins and develops the SAR response in ASM-treated plants

In silico Prediction and Validations of Domains Involved in Gossypium hirsutum SnRK1 Protein Interaction With Cotton Leaf Curl Multan Betasatellite Encoded βC1

Cotton leaf curl disease (CLCuD) caused by viruses of genus Begomovirus is a major constraint to cotton (Gossypium hirsutum) production in many cotton-growing regions of the world. Symptoms of the disease are caused by Cotton leaf curl Multan betasatellite (CLCuMB) that encodes a pathogenicity determinant protein, βC1. Here, we report the identification of interacting regions in βC1 protein by using computational approaches including sequence recognition, and binding site and interface prediction methods. We show the domain-level interactions based on the structural analysis of G. hirsutum SnRK1 protein and its domains with CLCuMB-βC1. To verify and validate the in silico predictions, three different experimental approaches, yeast two hybrid, bimolecular fluorescence complementation and pull down assay were used. Our results showed that ubiquitin-associated domain (UBA) and autoinhibitory sequence (AIS) domains of G. hirsutum-encoded SnRK1 are involved in CLCuMB-βC1 interaction. This is the first comprehensive investigation that combined in silico interaction prediction followed by experimental validation of interaction between CLCuMB-βC1 and a host protein. We demonstrated that data from computational biology could provide binding site information between CLCuD-associated viruses/satellites and new hosts that lack known binding site information for protein–protein interaction studies. Implications of these findings are discussed

IN VIVO LOCALIZATION AND INTERACTION STUDIES OF TOSPOVIRUS (GENUS: TOSPOVIRUS; FAMILY: BUNYAVIRIDAE) PROTEINS

IN VIVO LOCALIZATION AND INTERACTION STUDIES OF TOSPOVIRUS (GENUS: TOSPOVIRUS; FAMILY: BUNYAVIRIDAE) PROTEINSTospoviruses (family: Bunyaviridae) are one of the most agronomically important viruses that affect numerous agriculture crops. Two distinct tospoviruses, Tomato spotted wilt virus (TSWV) and Iris yellow spot virus (IYSV), are important pathogens of a wide variety of crops. Localization and interaction studies of IYSV proteins were performed in the host plant, Nicotiana benthamiana. Our localization studies in infected plants suggested a change in localization of the nucleocapsid (N) protein from the cytoplasm to cell periphery. The interaction analyses of tospovirus proteins were performed using Bimolecular Fluorescence Complementation (BiFC), pull down, and Yeast-2-Hybrid (Y2H) assays. Interaction studies of IYSV proteins demonstrated homotypic and heterotypic interactions between IYSV N and NSm proteins. The observations were discussed in the context of tospovirus assembly, replication, and infection processes.To understand the molecular basis of mixed infections caused by tospoviruses, interaction between N and NSm proteins of TSWV and IYSV was also studied. Results showed that the N and NSm proteins of IYSV interact with their counterparts coded by TSWV in a doubly infected plant. Interacting regions of these proteins were also identified by the yeast-2-hybrid systems. Molecular interactions between cognate proteins of these viruses may be of crucial significance for the understanding of pathogenesis, evolution, and development of efficient and durable control strategies for these viruses.Acibenzolar-S-Methyl (ASM) is a functional analog of salicylic acid (SA) that activates plant's local and systemic acquired resistance (SAR) responses against a wide variety of pathogens. Two different hosts of IYSV-Datura stramonium and Nicotiana benthamiana were used as model plants to develop a set of descriptors to study the effect of SAR inducers on IYSV infection. ASM- and buffer-treated D. stramonium and N. benthamiana plants were mechanically inoculated with IYSV. Symptom development and virus levels were monitored by the enzyme linked immunosorbent assay (ELISA), reverse-transcriptase polymerase chain reaction (RT-PCR), and real-time- RT-PCR (q-RT-PCR). A significant reduction in IYSV levels in ASM-treated plants was noticed. In addition, ASM-treated plants showed reduced viral symptoms (lesion number and sizes), compared to buffer-treated plants. The knowledge gained from this study could be used in developing SAR based chemical options for virus management

A crosstalk between extracellular ATP and jasmonate signaling pathways for plant defense

Damage-associated molecular patterns (DAMPs), such as extracellular ATP, act as danger signals in response to biotic and abiotic stresses. Extracellular ATP is perceived by a plant purinoceptor, P2 receptor kinase 1 (P2K1), inducing downstream signaling for defense responses. How ATP induces these defense responses has not been well studied. A recent study by Tripathi et al. (Plant Physiology, 176: 511-523, 2018) revealed a synergistic interaction between extracellular ATP and jasmonate (JA) signaling during plant defense responses. This signaling crosstalk requires the formation of secondary messengers, i.e., cytosolic calcium, reactive oxygen species, and nitric oxide. This finding has given a new direction towards understanding the defense signals activated by DAMPs. In this addendum, we discuss possible insights into how extracellular ATP signaling interacts with the JA signaling pathway for plant defense responses

Chemical elicitors of systemic acquired resistance—Salicylic acid and its functional analogs

Any interaction of plants with phytopathogens involves the generation of various chemical molecules that are critical for activation of their defense machinery. One of the chemicals, salicylic acid (SA) induces systemic acquired resistance (SAR) in plants. The activation of SAR provides a broad-spectrum resistance against a wide range of related or unrelated pathogens. There has been considerable progress in the biochemical and molecular understanding of SAR activation in various plants. In addition, several chemicals including SA and its analogs are known to provide a direct or indirect defense against pathogens when applied to plants. Molecular mechanism of plant defense induced by synthetic chemical inducers is not very well understood. This review highlights the importance of salicylic acid and its most studied analog, Acibenzolar-S-methyl in inducing SAR and it also provides a description of other major chemical elicitors of plant defenses and their possible molecular mechanism. Keywords: SA, SAR, Acibenzolar-S-methyl, Chemical elicitors, Defense signalin

A crosstalk between extracellular ATP and jasmonate signaling pathways for plant defense

Damage-associated molecular patterns (DAMPs), such as extracellular ATP, act as danger signals in response to biotic and abiotic stresses. Extracellular ATP is perceived by a plant purinoceptor, P2 receptor kinase 1 (P2K1), inducing downstream signaling for defense responses. How ATP induces these defense responses has not been well studied. A recent study by Tripathi et al. (Plant Physiology, 176: 511-523, 2018) revealed a synergistic interaction between extracellular ATP and jasmonate (JA) signaling during plant defense responses. This signaling crosstalk requires the formation of secondary messengers, i.e., cytosolic calcium, reactive oxygen species, and nitric oxide. This finding has given a new direction towards understanding the defense signals activated by DAMPs. In this addendum, we discuss possible insights into how extracellular ATP signaling interacts with the JA signaling pathway for plant defense responses

Movement and nucleocapsid proteins coded by two tospovirus species interact through multiple binding regions in mixed infections

Negative-stranded tospoviruses (family: Bunyaviridae) are among the most agronomically important viruses. Some of the tospoviruses are known to exist as mixed infections in the same host plant. Iris yellow spot virus (IYSV) and Tomato spotted wilt virus (TSWV) were used to study virus-virus interaction in dually infected host plants. Viral genes of both viruses were separately cloned into binary pSITE-BiFC vectors. BiFC results showed that the N and NSm proteins of IYSV interact with their counterparts coded by TSWV in dually infected Nicotiana benthamiana plants. BiFC results were further confirmed by pull down and yeast-2-hybrid (Y2H) assays. Interacting regions of the N and NSm proteins were also identified by Y2H system and β-galactosidase activity. Several regions of the N and NSm were found interacting with each other. The regions involved in these interactions are presumed to be critical for the functioning of the tospovirus N and NSm proteins. This is the first report of in vivo protein interactions of distinct tospoviruses in mixed infection

SABP2, a Methyl Salicylate Esterase Is Required for the Systemic Acquired Resistance Induced by Acibenzolar-S-methyl in Plants

Tobacco SABP2, a 29. kDa protein catalyzes the conversion of methyl salicylic acid (MeSA) into salicylic acid (SA) to induce SAR. Pretreatment of plants with acibenzolar-. S-methyl (ASM), a functional analog of salicylic acid induces systemic acquired resistance (SAR). Data presented in this paper suggest that SABP2 catalyzes the conversion of ASM into acibenzolar to induce SAR. Transgenic SABP2-silenced tobacco plants when treated with ASM, fail to express PR-1 proteins and do not induce robust SAR expression. When treated with acibenzolar, full SAR is induced in SABP2-silenced plants. These results show that functional SABP2 is required for ASM-mediated induction of resistance

Oxidative and Glycation Damage to Mitochondrial DNA and Plastid DNA during Plant Development

Oxidative damage to plant proteins, lipids, and DNA caused by reactive oxygen species (ROS) has long been studied. The damaging effects of reactive carbonyl groups (glycation damage) to plant proteins and lipids have also been extensively studied, but only recently has glycation damage to the DNA in plant mitochondria and plastids been reported. Here, we review data on organellar DNA maintenance after damage from ROS and glycation. Our focus is maize, where tissues representing the entire range of leaf development are readily obtained, from slow-growing cells in the basal meristem, containing immature organelles with pristine DNA, to fast-growing leaf cells, containing mature organelles with highly-fragmented DNA. The relative contributions to DNA damage from oxidation and glycation are not known. However, the changing patterns of damage and damage-defense during leaf development indicate tight coordination of responses to oxidation and glycation events. Future efforts should be directed at the mechanism by which this coordination is achieved

Extracellular ATP Acts on Jasmonate Signaling to Reinforce Plant Defense

Damaged cells send various signals to stimulate defense responses. Recent identification and genetic studies of the plant purinoceptor, P2K1 (also known as DORN1), have demonstrated that extracellular ATP is a signal involved in plant stress responses, including wounding, perhaps to evoke plant defense. However, it remains largely unknown how extracellular ATP induces plant defense responses. Here, we demonstrate that extracellular ATP induces plant defense mediated through activation of the intracellular signaling of jasmonate (JA), a well-characterized defense hormone. In Arabidopsis ( ) leaves, ATP pretreatment induced resistance against the necrotrophic fungus, The induced resistance was enhanced in the P2K1 receptor overexpression line, but reduced in the receptor mutant, - Mining the transcriptome data revealed that ATP induces a set of JA-induced genes. In addition, the P2K1-associated coexpression network contains defense-related genes, including those encoding jasmonate ZIM-domain (JAZ) proteins, which play key roles as repressors of JA signaling. We examined whether extracellular ATP impacts the stability of JAZ1 in Arabidopsis. The results showed that the JAZ1 stability decreased in response to ATP addition in a proteasome-dependent manner. This reduction required intracellular signaling via second messengers-cytosolic calcium, reactive oxygen species, and nitric oxide. Interestingly, the ATP-induced JAZ1 degradation was attenuated in the JA receptor mutant, , but not in the JA biosynthesis mutant, , or upon addition of JA biosynthesis inhibitors. Immunoprecipitation analysis demonstrated that ATP increases the interaction between COI1 and JAZ1, suggesting direct cross talk between extracellular ATP and JA in intracellular signaling events. Taken together, these results suggest that extracellular ATP signaling directly impacts the JA signaling pathway to maximize plant defense responses