Genome-wide analysis of the CalS gene family in cotton reveals their potential roles in fiber development and responses to stress

Callose deposition occurs during plant growth and development, as well as when plants are under biotic and abiotic stress. Callose synthase is a key enzyme for the synthesis of callose. In this study, 27, 28, 16, and 15 callose synthase family members were identified in Gossypium hirsutum, Gossypium barbadense, Gossypium raimondii, and Gossypium arboreum using the sequence of Arabidopsis callose synthase. The CalSs were divided into five groups by phylogenetic, gene structure, and conservative motif analysis. The conserved motifs and gene structures of CalSs in each group were highly similar. Based on the analysis of cis-acting elements, it is inferred that GhCalSs were regulated by abiotic stress. WGD/Segmental duplication promoted the amplification of the CalS gene in cotton, and purification selection had an important function in the CalS family. The transcriptome data and qRT-PCR under cold, heat, salt, and PEG treatments showed that GhCalSs were involved in abiotic stress. The expression patterns of GhCalSs were different in various tissues. We predicted that GhCalS4, which was highly expressed in fibers, had an important effect on fiber elongation. Hence, these results help us understand the role of GhCalSs in fiber development and stress response.

Biochemical and Cytological Interactions Between Callose Synthase and Microtubules in the Tobacco Pollen Tube

Callose is a cell wall polysaccharide involved in several fundamental biological processes, ranging from plant development to response to abiotic and biotic stresses. To understand how callose deposition is regulated, it is important to know how its synthesizing enzyme, i.e., callose synthase, is regulated and if it interacts with vesicular-cytoskeletal system of plant cells. Actin filaments are thought to determine the long-range distribution of callose synthase through transport vesicles. Unlike other enzymes (such as cellulose synthase) that synthesize cell wall polysaccharides, the spatial and biochemical relationships between callose synthase and microtubules are poorly understood. Some experimental evidence already support the association between callose synthase and tubulin, however, despite its importance in maintaining plant integrity, knowledge about regulation of callose biosynthesis is still limited. Here we investigated the association between callose synthase and cytoskeleton by biochemical and ultrastructural analyses in a model system, pollen tube, where callose is an essential cell wall component. Native 2-D electrophoresis and isolation of the callose synthase complex confirmed that callose synthase is associated with tubulin and can interface with cortical microtubules. In contrast, actin and sucrose synthase (which supplies UDP-glucose to callose synthase) are not permanently associated with callose synthase. Immunogold labeling showed strong colocalization of the enzyme and microtubules; this association is occasionally mediated by vesicles. The association between callose synthase and vesicles was also demonstrated by co-distribution between the enzyme and Rab11b; in addition, the not homogeneous distribution of callose synthase in cell membranes is also shown by analysis of membrane microdomains.

PMR4-dependent cell wall depositions are a consequence but not the cause of temperature-induced autoimmunity

Plants possess a well-balanced immune system that is required for defense against pathogen infections. In autoimmune mutants or necrotic crosses, an intrinsic temperature-dependent imbalance leads to constitutive immune activation, resulting in severe damage or even death of plants. Recently, cell wall depositions were described as one of the symptoms following induction of the autoimmune phenotype in Arabidopsis saul1-1 mutants. However, the regulation and function of these depositions remained unclear. Here, we show that cell wall depositions, containing lignin and callose, were a common autoimmune feature and were deposited in proportion to the severity of the autoimmune phenotype at reduced ambient temperatures. When plants were exposed to reduced temperature for periods insufficient to induce an autoimmune phenotype, the cell wall depositions were not present. After low temperature intervals, sufficient to induce autoimmune responses, cell wall depositions correlated with a point of no return in saul1-1 autoimmunity. Although cell wall depositions were largely abolished in saul1-1 pmr4-1 double mutants lacking SAUL1 and the callose synthase gene GSL5/PMR4, their phenotype remained unchanged compared to that of the saul1-1 single mutant. Our data showed that cell wall depositions generally occur in autoimmunity, but appear not to be the cause of autoimmune phenotypes.

Fungal F8-Culture Filtrate Induces Tomato Resistance against Tomato Yellow Leaf Curl Thailand Virus

Tomato (Solanum lycopersicum) is an important economic crop worldwide. However, tomato production is jeopardized by the devastating tomato yellow leaf curl disease (TYLCD) caused by whitefly-transmitted begomoviruses (WTBs). In this study, we evaluated the efficacy of our previously developed plant antiviral immunity inducer, fungal F8-culture filtrate, on tomato to combat tomato yellow leaf curl Thailand virus (TYLCTHV), the predominant WTB in Taiwan. Our results indicated that F8-culture filtrate treatment induced strong resistance, did not reduce the growth of tomato, and induced prominent resistance against TYLCTHV both in the greenhouse and in the field. Among TYLCTHV-inoculated Yu-Nu tomato grown in the greenhouse, a greater percentage of plants treated with F8-culture filtrate (43–100%) were healthy-looking compared to the H2O control (0–14%). We found that TYLCTHV cannot move systemically only on the F8-culture filtrate pretreated healthy-looking plants. Tracking the expression of phytohormone-mediated immune maker genes revealed that F8-culture filtrate mainly induced salicylic acid-mediated plant immunity. Furthermore, callose depositions and the expression of the pathogen-induced callose synthase gene, POWDERY MILDEW RESISTANT 4 were only strongly induced by TYLCTHV on tomato pretreated with F8-culture filtrate. This study provides an effective way to induce tomato resistance against TYLCTHV.

The receptor kinase SRF3 coordinates iron-level and flagellin dependent defense and growth responses in plants

Iron is critical for host-pathogen interactions. While pathogens seek to scavenge iron to spread, the host aims at decreasing iron availability to reduce pathogen virulence. Thus, iron sensing and homeostasis are of particular importance to prevent host infection and part of nutritional immunity. While the link between iron homeostasis and immunity pathways is well established in plants, how iron levels are sensed and integrated with immune response pathways remain unknown. We identified a receptor kinase, SRF3 coordinating root growth, iron homeostasis and immunity pathways via regulation of callose synthase activity. These processes are modulated by iron levels and rely on SRF3 extracellular and kinase domain which tune its accumulation and partitioning at the cell surface. Mimicking bacterial elicitation with the flagellin peptide flg22 phenocopies SRF3 regulation upon low iron levels and subsequent SRF3-dependent responses. We propose that SRF3 is part of nutritional immunity responses involved in sensing external iron levels.

The receptor kinase SRF3 coordinates iron-level and flagellin dependent defense and growth responses in plants

Iron is critical for host-pathogen interactions. While pathogens seek to scavenge iron to spread, the host aims at decreasing iron availability to reduce pathogen virulence. Thus, iron sensing and homeostasis are of particular importance to prevent host infection and part of nutritional immunity. While the link between iron homeostasis and immunity pathways is well established in plants, how iron levels are sensed and integrated with immune response pathways remain unknown. We identified a receptor kinase, SRF3 coordinating root growth, iron homeostasis and immunity pathways via regulation of callose synthase activity. These processes are modulated by iron levels and rely on SRF3 extracellular and kinase domain which tune its accumulation and partitioning at the cell surface. Mimicking bacterial elicitation with the flagellin peptide flg22 phenocopies SRF3 regulation upon low iron levels and subsequent SRF3-dependent responses. We propose that SRF3 is part of nutritional immunity responses involved in sensing external iron levels.

Callose-synthesizing enzymes as membrane proteins of Betula protoplasts secrete bundles of β-1,3-glucan hollow fibrils under Ca2+-rich and acidic culture conditions

Previously, it was reported that plant protoplasts isolated from Betula platyphylla (white birch) callus secreted bundles of hollow callose fibrils in acidic culture medium containing a high concentration of calcium ions (Ca2+). Here, the callose synthase was characterized from in situ and in vitro perspectives. Localization of callose synthases at the secreting site of callose fiber was indicated from in situ immunostaining observation of protoplasts. For in vitro analyses, membrane proteins were extracted from membrane fraction of protoplasts with a 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate (CHAPS) treatment. The CHAPS extract aggregated in the presence of a high concentration of Ca2+, suggesting that Ca2+ may promote the arrangement of callose synthases in the plasma membrane. The callose synthase activity was dependent on pH and Ca2+, similar to the callose synthase of Arabidopsis thaliana. However, the synthesized fibril products were longer than those produced by callose synthases of herbaceous plants. This is the first insight into the specific properties of callose synthases of woody plants that secrete of callose hollow fibers.

Characterization of Callose Deposition and Analysis of the Callose Synthase Gene Family of Brassica napus in Response to Leptosphaeria maculans

Callose plays a critical role in different biological processes including development as well as in the response to multiple biotic and abiotic stresses. In this study, we characterized the callose deposition in cotyledons of different Brassica napus varieties post-inoculated with different Leptosphaeria maculans isolates. Further, members of the callose synthase gene were identified from the whole genome of B. napus using the 12 Arabidopsis thaniana callose synthase protein sequences, and were then classified into three groups based on their phylogenetic relationships. Chromosomal location and duplication patterns indicated uneven distribution and segmental duplication patterns of BnCalS genes in the B. napus genome. Subsequently, gene structures, conserved domains analysis, and protein properties were analyzed for BnCalS genes. In addition, 12 B. napus orthologs of the AtCalS were selected for investigating the tissue expression pattern, indicating diverse expression patterns for these BnCalS genes. Responses of the selected 12 orthologs and all the BnCalS genes were characterized in the different types (AvrLm1-Rlm1, AvrLm4-Rlm4, AvrLepR1-LepR1) of B. napus–L. maculans interactions and B. napus-Leptosphaeria biglobosa interactions, implying their potential roles in response to Leptosphaeria infection.