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Background: Zinc finger proteins (ZFPs) are important regulators in abiotic and biotic stress tolerance in plants. However, the role of the ZFPs in wheat responding to pathogen infection is poorly understood. Results: In this study, we found TaZFP8-5B was down-regulated by Puccinia striiformis f. sp. tritici (Pst) infection. TaZFP8-5B possesses a single C2H2-type zinc finger domain with a plant-specific QALGGH motif, and an EAR motif (LxLxL) at the C-terminus. The EAR motif represses the trans-activation ability of TaZFP8-5B. Knocking down the expression of TaZFP8 by virus-induced gene silencing increased wheat resistance to Pst, whereas TaZFP8-5B-overexpressing reduced wheat resistance to stripe rust and rice resistance to Magnaporthe oryzae, suggesting that TaZFP8-5B plays a negative role in the modulation of plant immunity. Using bimolecular fluorescence complementation, split-luciferase, and yeast two-hybrid assays, we showed that TaZFP8-5B interacted with a wheat calmodulin-like protein TaCML21. Knock-down of TaCML21 reduced wheat resistance to Pst. Conclusions: This study characterized the function of TaZFP8-5B and its interacting protein TaCML21. Our findings provide a new perspective on a regulatory module made up of TaCML21-TaZFP8-5B in plant immunity. [ABSTRACT FROM AUTHOR]

Calmodulins (CaMs) and calmodulin-like proteins (CMLs) belong to families of calcium-sensors that act as calcium ion (Ca2+) signal-decoding proteins and regulate downstream target proteins. As a tropical halophyte, Canavalia rosea shows great resistance to multiple abiotic stresses, including high salinity/alkalinity, extreme drought, heat, and intense sunlight. However, investigations of calcium ion signal transduction involved in the stress responses of C. rosea are limited. The CaM and CML gene families have been identified and characterized in many other plant species. Nevertheless, there is limited available information about these genes in C. rosea. In this study, a bioinformatic analysis, including the gene structures, conserved protein domains, phylogenetic relationships, chromosome distribution, and gene synteny, was comprehensively performed to identify and characterize CrCaMs and CrCMLs. A spatio-temporal expression assay in different organs and environmental conditions was then conducted using the RNA sequencing technique. Additionally, several CrCaM and CrCML members were then cloned and functionally characterized using the yeast heterogeneous expression system, and some of them were found to change the tolerance of yeast to heat, salt, alkalinity, and high osmotic stresses. The results of this study provide a foundation for understanding the possible roles of the CrCaM and CrCML genes, especially for halophyte C. rosea's natural ecological adaptability for its native habitats. This study also provides a theoretical basis for further study of the physiological and biochemical functions of plant CaMs and CMLs that are involved in tolerance to multiple abiotic stresses. [ABSTRACT FROM AUTHOR]

Plant calmodulins (CaMs) and calmodulin-like proteins (CMLs) mediate Ca2+ signaling in response to abiotic stresses. Manipulation of this signaling in crops could increase stress tolerance. We review methods for detecting Ca2+ signals, regulatory roles of CaMs and CMLs, binding targets, and Ca2+ networks under abiotic stress in organelles. [ABSTRACT FROM AUTHOR]

Abstract Background Zinc finger proteins (ZFPs) are important regulators in abiotic and biotic stress tolerance in plants. However, the role of the ZFPs in wheat responding to pathogen infection is poorly understood. Results In this study, we found TaZFP8-5B was down-regulated by Puccinia striiformis f. sp. tritici (Pst) infection. TaZFP8-5B possesses a single C2H2-type zinc finger domain with a plant-specific QALGGH motif, and an EAR motif (LxLxL) at the C-terminus. The EAR motif represses the trans-activation ability of TaZFP8-5B. Knocking down the expression of TaZFP8 by virus-induced gene silencing increased wheat resistance to Pst, whereas TaZFP8-5B-overexpressing reduced wheat resistance to stripe rust and rice resistance to Magnaporthe oryzae, suggesting that TaZFP8-5B plays a negative role in the modulation of plant immunity. Using bimolecular fluorescence complementation, split-luciferase, and yeast two-hybrid assays, we showed that TaZFP8-5B interacted with a wheat calmodulin-like protein TaCML21. Knock-down of TaCML21 reduced wheat resistance to Pst. Conclusions This study characterized the function of TaZFP8-5B and its interacting protein TaCML21. Our findings provide a new perspective on a regulatory module made up of TaCML21-TaZFP8-5B in plant immunity.

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Summary: It is well known that calcium, ethylene and abscisic acid (ABA) can regulate fruit ripening, however, their interaction in the regulation of fruit ripening has not yet been fully clarified. The present study found that the expression of the papaya calcium sensor CpCML15 was strongly linked to fruit ripening. CpCML15 could bind Ca2+ and served as a true calcium sensor. CpCML15 interacted with CpPP2C46 and CpPP2C65, the candidate components of the ABA signalling pathways. CpPP2C46/65 expression was also related to fruit ripening and regulated by ethylene. CpCML15 was located in the nucleus and CpPP2C46/65 were located in both the nucleus and membrane. The interaction between CpCML15 and CpPP2C46/65 was calcium dependent and further repressed the activity of CpPP2C46/65 in vitro. The transient overexpression of CpCML15 and CpPP2C46/65 in papaya promoted fruit ripening and gene expression related to ripening. The reduced expression of CpCML15 and CpPP2C46/65 by virus‐induced gene silencing delayed fruit colouring and softening and repressed the expression of genes related to ethylene signalling and softening. Moreover, ectopic overexpression of CpCML15 in tomato fruit also promoted fruit softening and ripening by increasing ethylene production and enhancing gene expression related to ripening. Additionally, CpPP2C46 interacted with CpABI5, and CpPP2C65 interacted with CpERF003‐like, two transcriptional factors in ABA and ethylene signalling pathways that are closely related to fruit ripening. Taken together, our results showed that CpCML15 and CpPP2Cs positively regulated fruit ripening, and their interaction integrated the cross‐talk of calcium, ABA and ethylene signals in fruit ripening through the CpCML15‐CpPP2Cs‐CpABI5/CpERF003‐like pathway. [ABSTRACT FROM AUTHOR]

The root‐knot nematode Meloidogyne graminicola secretes effectors into rice tissues to modulate host immunity. Here, we characterised MgCRT1, a calreticulin protein of M. graminicola, and identified its target in the plant. In situ hybridisation showed MgCRT1 mRNA accumulating in the subventral oesophageal gland in J2 nematodes. Immunolocalization indicated MgCRT1 localises in the giant cells during parasitism. Host‐induced gene silencing of MgCRT1 reduced the infection ability of M. graminicola, while over‐expressing MgCRT1 enhanced rice susceptibility to M. graminicola. A yeast two‐hybrid approach identified the calmodulin‐like protein OsCML31 as an interactor of MgCRT1. OsCML31 interacts with the high mobility group protein OsHMGB1 which is a conserved DNA binding protein. Knockout of OsCML31 or overexpression of OsHMGB1 in rice results in enhanced susceptibility to M. graminicola. In contrast, overexpression of OsCML31 or knockout of OsHMGB1 in rice decreases susceptibility to M. graminicola. The GST‐pulldown and luciferase complementation imaging assay showed that MgCRT1 decreases the interaction of OsCML31 and OsHMGB1 in a competitive manner. In conclusion, when M. graminicola infects rice and secretes MgCRT1 into rice, MgCRT1 interacts with OsCML31 and decreases the association of OsCML31 with OsHMGB1, resulting in the release of OsHMGB1 to enhance rice susceptibility. Summary statement: A plant‐pathogenic nematode effector MgCRT1 interacts with OsCML31 and decreases the association of OsCML31 with OsHMGB1 in a competitive manner, resulting in the release of OsHMGB1 to enhance rice susceptibility. [ABSTRACT FROM AUTHOR]

Summary: Transient and rapid increase in cytosolic Ca2+ plays a crucial role in plant–pathogen‐associated molecular pattern (PAMP)‐triggered immunity (PTI). Cyclic nucleotide‐gated channels (CNGCs) have been implicated in mediating this Ca2+ influx; however, their regulatory mechanisms remain poorly understood.Here, we have found that AVRblb2 requires the calmodulin (CaM) and calmodulin‐like (CML) proteins as co‐factors to interact with the NbCNGCs, resulting in the formation of AVRblb2‐CaM/CML‐NbCNGCs complex. Furthermore, CaM and CML are dissociated from NbCNGC18 during PTI response to increase Ca2+ influx; however, Avrblb2 inhibits calcium channel activation by disrupting the release of CaM and CML from NbCNGC18.Following recognition of PAMP, NbCNGC18 forms active heteromeric channels with other NbCNGCs, which may give selectivity of CNGC complex against diverse signals for fine‐tuning of cytosolic Ca2+ level to mediate appropriate responses. Silencing of multiple NbCNGCs compromised the function of AVRblb2 on the pathogenicity of Phytophthora infestans, confirming that AVRblb2 contributes to pathogen virulence by targeting CNGCs.Our findings provide new insights into the regulation of CNGCs in PTI and the role of pathogen effectors in manipulating host cell physiology to promote infection. [ABSTRACT FROM AUTHOR]

Calcium signaling plays a crucial role as a second messenger in plant immune responses. However, the regulatory mechanisms that govern host responses to infections caused by Sclerotinia sclerotiorum and Phytophthora capsici remain unclear. This study investigated the expression pattern of Nicotiana benthamiana calmodulin-like protein 30 (NbCML30) following infections. Our results reveal a significant downregulation of NbCML30 expression after S. sclerotiorum infection, with no changes observed in NbCML30 expression following P. capsici infection. Importantly, transient silencing of NbCML30 using tobacco rattle virus-mediated methods promoted infections caused by both S. sclerotiorum and P. capsici. In contrast, transgenic plants overexpressing NbCML30 exhibited a suppressive effect on the infections caused by S. sclerotiorum and P. capsici. Subsequent transcriptome sequencing uncovered that NbCML30 silencing markedly upregulated the expression of host endogenous genes, particularly activating pathways associated with protein processing in endoplasmic reticulum (ER) associated degradation (ERAD) pathway. These findings highlight the affirmative regulatory role of NbCML30 in conferring resistance against necrotrophic and hemibiotrophic pathogens, shedding light on its potential inhibitory mechanism. In conclusion, our study provides a theoretical basis for crop resistance genetic breeding strategies by elucidating the regulatory function of NbCML30 in plant defense against these pathogens. • Silencing NbCML30 enhances susceptibility to S. sclerotiorum and P. capsici infections. • Silencing NbCML30 activates pathways associated with endoplasmic reticulum-associated degradation (ERAD). • NbCML30 overexpression inhibits S. sclerotiorum and P. capsici infection. [ABSTRACT FROM AUTHOR]

The effect of calcium chloride (CaCl 2) treatment on γ-aminobutyric acid (GABA) accumulation in fresh-cut cantaloupe and the involved mechanisms were investigated. The result showed that 1% (w/v) CaCl 2 treatment increased GABA content and activities of glutamate decarboxylase (GAD) and succinate semialdehyde dehydrogenase (SSADH), while decreased glutamate (Glu) content and GABA transaminase (GABA-T) activities in fresh-cut cantaloupe. CmCML11 and CmCAMTA5 expressions of CaCl 2 -treated fruit increased by 187.4% and 165.6% than control fruit in the initial 6 h. Besides, expressions of GABA shunt genes, including CmGAD1 , CmGAD2 , CmGABA-T and CmSSADH were also up-regulated by CaCl 2 treatment during early storage. Moreover, acting as a transcriptional activator, CmCAMTA5 could bind to the CG-box in promoters of CmGAD1 , CmGABA-T and CmSSADH and activate their transcription. Furthermore, the interaction between CmCML11 and CmCAMTA5 could enhance the transcriptional activation on GABA shunt genes which were regulated by CmCAMTA5. Collectively, our findings revealed that CaCl 2 treatment promoted GABA accumulation in fresh-cut cantaloupe via the combined effect of CmCML11 and CmCAMTA5 in the regulation of expressions of CmGAD1 , CmGABA-T , and CmSSADH in GABA shunt. [Display omitted] • CaCl 2 treatment could activate GABA shunt and enhance GABA content of fresh-cut cantaloupe. • CaCl 2 treatment up-regulated the expressions of CmCML11 and CmCAMTA5. • CmCML11 protein physically interacted with CmCAMTA5. • GABA shunt was positively regulated by Ca2+-CML11-CAMTA5 signal transduction pathway. [ABSTRACT FROM AUTHOR]