The host immune response against powdery mildew was tested in H. brasiliensis. 
Supplementary Figure 1 | Determinations of ROS accumulation and HR in Nicotiana benthamiana leaves. 16 Articles, This article is part of the Research Topic, https://doi.org/10.3389/fmicb.2020.591387, https://www.frontiersin.org/articles/10.3389/fmicb.2020.591387/full#supplementary-material, Creative Commons Attribution License (CC BY). Container-grown rubber plants require frequent watering, combined with humidity and cool temperatures, which can result in edema. From each sample, a 5 g of protein was analyzed by SDS-PAGE and western blotting. Epoxycarotenoid cleavage by NCED5 fine-tunes ABA accumulation and affects seed dormancy and drought tolerance with other NCED family members. Powdery mildews are characterized by contracted carbohydrate metabolism and diverse effectors to adapt to obligate biotrophic lifestyle. When secreted effectors are recognized by the host resistance gene products, another layer of immunity is triggered, known as effector-triggered immunity (ETI). Overfertilization can cause a buildup of salts in plants, which can burn their young roots. Rubber trees (Hevea brasiliensis) are the primary source of natural rubber, and powdery mildew infection causes significant losses to natural rubber yields. Meanwhile, if you notice brown or white-like corky lumps, the plant may be suffering from oedema. It is essential to check your plant thoroughly for any indications of white spots. Foliage is dark green.
The results indicated that the interacting proteins of EqCSEP01276 included HbNCED5, one of the key enzymes in ABA biosynthesis, and two other uncharacterized proteins (Supplementary Table 2). }, 31960518 and 31660033), National Key R&D Program of China (No. 19:3900. doi: 10.3390/ijms19123900, Mei, S., Hou, S., Cui, H., Feng, F., and Rong, W. (2016). Annu. HbNCED5 contains a putative N-terminal chloroplast transit peptide (aa 149), which was analyzed by ChloroP 1.1 Server3 (Supplementary Table 3), and HbNCED5 probably has a chloroplast localization. PLoS Pathog. (2016).
Thus, the sharing of signaling proteins possibly connects different hormone pathways (Spoel et al., 2003, 2009; Tada et al., 2008). It also induced high expression of defense-related enzyme genes, including PR-1, and heavy depositions of callose and lignin. 1630042020008). In H. brasiliensis, foliar application of chitosan used as an elicitor enhances resistance to Phytophthora disease by inducing higher levels of ABA. 8:719. doi: 10.3389/fpls.2017.00719, Kuyyogsuy, A., Deenamo, N., Khompatara, K., Ekchaweng, K., and Churngchow, N. J. P. (2018). We observed that in cells in the presence of strong EqCSEP01276SP-GFP signals, the intensity of HbNCED5-RFP with punctate distribution was reduced (Figures 9A,B). Opin. Callose depositions were visualized by aniline blue staining. (2017). "acceptedAnswer": { Do not let oedema develop in your rubber plant by maintaining an environment that does not favor the condition. Another way to promote warmth is through the use of artificial lights. It is a natural process of your rubber plant, so dont panic about it. GFP (emission wavelength: 488 nm, excitation wavelength: 680 nm), RFP (emission wavelength: 558 nm, excitation wavelength: 583 nm), chlorophyll (emission wavelength: 470 nm, excitation wavelength: 680 nm), YFP (emission wavelength: 513 nm, excitation wavelength: 527 nm), and aniline blue (emission wavelength: 665 nm, excitation wavelength: 600 nm) were detected. Frost can cause devastating damage to your rubber plant. I am constantly looking for new houseplants to gradually transform my apartment into an urban jungle and grow veggies in my garden year-round. Always promote good airflow and ensure that your plant gets enough sunlight. If your rubber plant is heavily infected, it can be hard to get rid of powdery mildew, so prevent the disease from spreading further to your other houseplants. 168, 321333. A., Wright, C. M., Brodsky, J. L., and Greenberg, J. T. (2007). However, in Arabidopsis, mutants with defective ABA synthesis or perception showed increased resistance to Pseudomonas syringae, and ABA pretreatment increased susceptibility to the bacteria (de Torres-Zabala et al., 2007). 35, 24682483. } Use water-soluble fertilizer for your rubber plant and apply it only once a month. 70, 501512. This site is a participant in the Amazon Services LLC Associates Program, an affiliate advertising program designed to provide a means for sites to earn advertising fees by advertising and linking to Amazon.com. You can also use mouthwash to treat the white spots caused due to powdery mildew. Rubber tree leaves were infiltrated with 0.1% DAB solution for 10 h in the dark and boiled in 95% ethanol for 30 min to remove chlorophyll. In rubber plants, the development of white spots, along with other symptoms, is among the telltale signs of nutrient deficiency. Youll also notice that the parts of the plant are not growing in their standard sizes. Prevent dealing with powdery mildew by ensuring that your rubber plant is receiving enough light. Microb. We showed that foliar application of ABA in H. brasiliensis inhibited powdery mildew conidiation and growth and increased callose depositions in N. benthamiana. The EqCSEP01276 signal peptide was fused into the pSUC2 vector and was transformed into the yeast YTK12 strain. The plant develops white spots on the leaves if it is placed in direct sunlight. Bars = 50 m. 46, 220226. The end result of this is burned or scorched leaves and impeded growth. These results suggest that EqCSEP01276 is a secreted protein, and its signal peptide is required for its secretion. Safeguard your plant from freezing circumstances by wrapping them with plastic sheets, drop cloths, bed sheets, or even blankets. (B) In yeast two-hybrid assay, yeast transformants expressing EqCSEP01276SP and HbNCED5 or HbNCED were assayed for growth on SD-LW or SD-LWHA. Biol. Commun. YFP signals were detected in the N. benthamiana leaves transiently co-expressing EqCSEP01276SP-NYFP and HbNCED5-CYFP using confocal microscopy. SP, signal peptide. Natl. Thick, cotton-type wax on plants, on the other hand, can suggest your rubber plants has been infected with mealybugs. Remove the excess fertilizer on the soil of your rubber plant. 17, 13311343. (2016). (2019).
As oedema usually happens due to reduced transpiration on the surfaces of the leaves and a high level of soil moisture, you need to cut back on watering your rubber plant.
(B) The total leaf proteins and proteins from cytosol and chloroplasts were extracted from N. benthamiana leaves expressing EqCSEP01276SP-GFP. Three independent replicates with three areas per replicate were examined. Oedema is a disorder due to a physiological imbalance in the water of your plant. The editor and reviewer's affiliations are the latest provided on their Loop research profiles and may not reflect their situation at the time of review. This is me and my Chinese Money Plant.
As is the case with humans, nutrient deficiencies can cause serious trouble for rubber plants also. Furthermore, we introduced vectors into mesophyll protoplasts derived from H. brasiliensis leaves expressing EqCSEP01276SP-GFP because transient gene expression in H. brasiliensis protoplasts is feasible (Zhang et al., 2016). Plant immunity: towards an integrated view of plant-pathogen interactions. PLoS One 8:e73091. We assayed the secretory function of the EqCSEP01276 signal peptide using the yeast secretion system. The molecular mechanisms of E. quercicola infection have been understudied, partly because of the inability to culture this pathogen in an artificial medium. The E3 ligase APIP10 connects the effector AvrPiz-t to the NLR receptor piz-t in rice. This has made the development of suitable disease control strategies challenging. Expression of genes of interest was confirmed by Reversed Transcript PCR (RT-PCR). Pests may have laid eggs in the hidden parts of the plant and they may not be visible to the naked eye. Future efforts to assess this hypothesis are warranted. (2012). doi: 10.1007/978-1-4939-0799-1_4, Hansen, H., and Grossmann, K. (2000).
The pellet was collected, washed, and resuspended in distilled sterile water, and an aliquot was incubated at 37C for 10 min. Additionally, some EqCSEP01276SP-GFP signals were present in chloroplasts. Phytopathol. doi: 10.1146/annurev.phyto.050908.135202, Wu, H., Pan, Y., Di, R., He, Q., Rajaofera, M. J. N., Liu, W., et al. doi: 10.1016/j.micpath.2017.01.054, Lu, X., Kracher, B., Saur, I. M., Bauer, S., Ellwood, S. R., Wise, R., et al. Microb. When it comes to preventing diseases among houseplants, sanitation is key! Chloroplasts at the crossroad of photosynthesis, pathogen infection and plant defense. doi: 10.1016/j.biochi.2016.06.006, Bourras, S., McNally, K. E., Ben-David, R., Parlange, F., Roffler, S., Praz, C. R., et al. Figure 1. (A) Co-immunoprecipitation for interaction between EqCSEP01276SP and HbNCED5 or HbNCED.
The representative images were captured at 48 h post-inoculation. This makes sure that it dries properly by the end of the day. In rubber plants, the appearance of white spots is among the most significant signs of nutrient deficiency. Accumulation of ROS was visualized by staining the leaves with 3,3-diaminobenzidine (DAB). Meanwhile, GFP alone was used as a control to remove the non-specific proteins bound to the GFP-tag. Mix half a teaspoon of non-detergent soap and one tablespoon baking soda in a gallon of water. doi: 10.1104/pp.15.00278, PubMed Abstract | CrossRef Full Text | Google Scholar, Apel, K., and Hirt, H. (2004). Mean values are indicated by . Median values are indicated by the middle line. For callose deposition visualization, rubber tree or N. benthamiana leaves were washed with double distilled water (ddH2O) and soaked in 95% ethanol, followed by boiling in water to clear the chlorophyll. The resulting proteins were then analyzed by western blot using anti-GFP, anti-RFP, anti--actin, and anti-Rubisco antibodies. Additionally, bacterial pathogens also secrete numerous effectors by the type III secretion system to target host plant plastids, including chloroplasts.
Similar to the Avr1b signal peptide, the EqCSEP01276 signal peptide, but not the empty vector, allowed YTK12 to grow on the YPRAAA medium (Figure 4). It is an indication that there is a problem that you need to address immediately. 15:e1007620. It is identical to white spots on your leaves. The most common pests are mealy bugs and spider mites. Plant Cell Physiol. If aphids have infected your rubber plant, you can squish them with your fingers or simply prune the infested areas. WM and XL planned and designed the research. Individual values obtained from nine replicates are indicated by dots. We found that callose deposition in N. benthamiana was induced by the infiltration of 100 and 150 M ABA into the leaves (Supplementary Figures 3A,B). You can put your plant near your windows, but be sure that you are filtering the sunlight. Powdery mildew infects a wide range of agricultural, economic, and ornamental plants, causing significant losses. You can also use baby wipes or cotton swabs with alcohol to wipe the dust. Curr. As the leaves of rubber plants are dark green in color, you may think that it is easy to spot any white spots. doi: 10.21769/bioprotoc.1257, Liyanage, K. K., Khan, S., Brooks, S., Mortimer, P. E., Karunarathna, S. C., Xu, J., et al. Asterisks indicate significant differences (P < 0.01). Plants suffer further damage due to dehydration when the temperature warms up. It has been reported that the murine p53 protein domain forms a complex with the 40 large T-antigen (Tan et al., 1986). J. Proteome Res. Individual values (n = 9) are indicated by dots.
Supplementary Figure 1 | Determinations of ROS accumulation and HR in Nicotiana benthamiana leaves. 16 Articles, This article is part of the Research Topic, https://doi.org/10.3389/fmicb.2020.591387, https://www.frontiersin.org/articles/10.3389/fmicb.2020.591387/full#supplementary-material, Creative Commons Attribution License (CC BY). Container-grown rubber plants require frequent watering, combined with humidity and cool temperatures, which can result in edema. From each sample, a 5 g of protein was analyzed by SDS-PAGE and western blotting. Epoxycarotenoid cleavage by NCED5 fine-tunes ABA accumulation and affects seed dormancy and drought tolerance with other NCED family members. Powdery mildews are characterized by contracted carbohydrate metabolism and diverse effectors to adapt to obligate biotrophic lifestyle. When secreted effectors are recognized by the host resistance gene products, another layer of immunity is triggered, known as effector-triggered immunity (ETI). Overfertilization can cause a buildup of salts in plants, which can burn their young roots. Rubber trees (Hevea brasiliensis) are the primary source of natural rubber, and powdery mildew infection causes significant losses to natural rubber yields. Meanwhile, if you notice brown or white-like corky lumps, the plant may be suffering from oedema. It is essential to check your plant thoroughly for any indications of white spots. Foliage is dark green. The results indicated that the interacting proteins of EqCSEP01276 included HbNCED5, one of the key enzymes in ABA biosynthesis, and two other uncharacterized proteins (Supplementary Table 2). }, 31960518 and 31660033), National Key R&D Program of China (No. 19:3900. doi: 10.3390/ijms19123900, Mei, S., Hou, S., Cui, H., Feng, F., and Rong, W. (2016). Annu. HbNCED5 contains a putative N-terminal chloroplast transit peptide (aa 149), which was analyzed by ChloroP 1.1 Server3 (Supplementary Table 3), and HbNCED5 probably has a chloroplast localization. PLoS Pathog. (2016).
Thus, the sharing of signaling proteins possibly connects different hormone pathways (Spoel et al., 2003, 2009; Tada et al., 2008). It also induced high expression of defense-related enzyme genes, including PR-1, and heavy depositions of callose and lignin. 1630042020008). In H. brasiliensis, foliar application of chitosan used as an elicitor enhances resistance to Phytophthora disease by inducing higher levels of ABA. 8:719. doi: 10.3389/fpls.2017.00719, Kuyyogsuy, A., Deenamo, N., Khompatara, K., Ekchaweng, K., and Churngchow, N. J. P. (2018). We observed that in cells in the presence of strong EqCSEP01276SP-GFP signals, the intensity of HbNCED5-RFP with punctate distribution was reduced (Figures 9A,B). Opin. Callose depositions were visualized by aniline blue staining. (2017). "acceptedAnswer": { Do not let oedema develop in your rubber plant by maintaining an environment that does not favor the condition. Another way to promote warmth is through the use of artificial lights. It is a natural process of your rubber plant, so dont panic about it. GFP (emission wavelength: 488 nm, excitation wavelength: 680 nm), RFP (emission wavelength: 558 nm, excitation wavelength: 583 nm), chlorophyll (emission wavelength: 470 nm, excitation wavelength: 680 nm), YFP (emission wavelength: 513 nm, excitation wavelength: 527 nm), and aniline blue (emission wavelength: 665 nm, excitation wavelength: 600 nm) were detected. Frost can cause devastating damage to your rubber plant. I am constantly looking for new houseplants to gradually transform my apartment into an urban jungle and grow veggies in my garden year-round. Always promote good airflow and ensure that your plant gets enough sunlight. If your rubber plant is heavily infected, it can be hard to get rid of powdery mildew, so prevent the disease from spreading further to your other houseplants. 168, 321333. A., Wright, C. M., Brodsky, J. L., and Greenberg, J. T. (2007). However, in Arabidopsis, mutants with defective ABA synthesis or perception showed increased resistance to Pseudomonas syringae, and ABA pretreatment increased susceptibility to the bacteria (de Torres-Zabala et al., 2007). 35, 24682483. } Use water-soluble fertilizer for your rubber plant and apply it only once a month. 70, 501512. This site is a participant in the Amazon Services LLC Associates Program, an affiliate advertising program designed to provide a means for sites to earn advertising fees by advertising and linking to Amazon.com. You can also use mouthwash to treat the white spots caused due to powdery mildew. Rubber tree leaves were infiltrated with 0.1% DAB solution for 10 h in the dark and boiled in 95% ethanol for 30 min to remove chlorophyll. In rubber plants, the development of white spots, along with other symptoms, is among the telltale signs of nutrient deficiency. Youll also notice that the parts of the plant are not growing in their standard sizes. Prevent dealing with powdery mildew by ensuring that your rubber plant is receiving enough light. Microb. We showed that foliar application of ABA in H. brasiliensis inhibited powdery mildew conidiation and growth and increased callose depositions in N. benthamiana. The EqCSEP01276 signal peptide was fused into the pSUC2 vector and was transformed into the yeast YTK12 strain. The plant develops white spots on the leaves if it is placed in direct sunlight. Bars = 50 m. 46, 220226. The end result of this is burned or scorched leaves and impeded growth. These results suggest that EqCSEP01276 is a secreted protein, and its signal peptide is required for its secretion. Safeguard your plant from freezing circumstances by wrapping them with plastic sheets, drop cloths, bed sheets, or even blankets. (B) In yeast two-hybrid assay, yeast transformants expressing EqCSEP01276SP and HbNCED5 or HbNCED were assayed for growth on SD-LW or SD-LWHA. Biol. Commun. YFP signals were detected in the N. benthamiana leaves transiently co-expressing EqCSEP01276SP-NYFP and HbNCED5-CYFP using confocal microscopy. SP, signal peptide. Natl. Thick, cotton-type wax on plants, on the other hand, can suggest your rubber plants has been infected with mealybugs. Remove the excess fertilizer on the soil of your rubber plant. 17, 13311343. (2016). (2019).
As oedema usually happens due to reduced transpiration on the surfaces of the leaves and a high level of soil moisture, you need to cut back on watering your rubber plant.
(B) The total leaf proteins and proteins from cytosol and chloroplasts were extracted from N. benthamiana leaves expressing EqCSEP01276SP-GFP. Three independent replicates with three areas per replicate were examined. Oedema is a disorder due to a physiological imbalance in the water of your plant. The editor and reviewer's affiliations are the latest provided on their Loop research profiles and may not reflect their situation at the time of review. This is me and my Chinese Money Plant. As is the case with humans, nutrient deficiencies can cause serious trouble for rubber plants also. Furthermore, we introduced vectors into mesophyll protoplasts derived from H. brasiliensis leaves expressing EqCSEP01276SP-GFP because transient gene expression in H. brasiliensis protoplasts is feasible (Zhang et al., 2016). Plant immunity: towards an integrated view of plant-pathogen interactions. PLoS One 8:e73091. We assayed the secretory function of the EqCSEP01276 signal peptide using the yeast secretion system. The molecular mechanisms of E. quercicola infection have been understudied, partly because of the inability to culture this pathogen in an artificial medium. The E3 ligase APIP10 connects the effector AvrPiz-t to the NLR receptor piz-t in rice. This has made the development of suitable disease control strategies challenging. Expression of genes of interest was confirmed by Reversed Transcript PCR (RT-PCR). Pests may have laid eggs in the hidden parts of the plant and they may not be visible to the naked eye. Future efforts to assess this hypothesis are warranted. (2012). doi: 10.1007/978-1-4939-0799-1_4, Hansen, H., and Grossmann, K. (2000).
The pellet was collected, washed, and resuspended in distilled sterile water, and an aliquot was incubated at 37C for 10 min. Additionally, some EqCSEP01276SP-GFP signals were present in chloroplasts. Phytopathol. doi: 10.1146/annurev.phyto.050908.135202, Wu, H., Pan, Y., Di, R., He, Q., Rajaofera, M. J. N., Liu, W., et al. doi: 10.1016/j.micpath.2017.01.054, Lu, X., Kracher, B., Saur, I. M., Bauer, S., Ellwood, S. R., Wise, R., et al. Microb. When it comes to preventing diseases among houseplants, sanitation is key! Chloroplasts at the crossroad of photosynthesis, pathogen infection and plant defense. doi: 10.1016/j.biochi.2016.06.006, Bourras, S., McNally, K. E., Ben-David, R., Parlange, F., Roffler, S., Praz, C. R., et al. Figure 1. (A) Co-immunoprecipitation for interaction between EqCSEP01276SP and HbNCED5 or HbNCED.
The representative images were captured at 48 h post-inoculation. This makes sure that it dries properly by the end of the day. In rubber plants, the appearance of white spots is among the most significant signs of nutrient deficiency. Accumulation of ROS was visualized by staining the leaves with 3,3-diaminobenzidine (DAB). Meanwhile, GFP alone was used as a control to remove the non-specific proteins bound to the GFP-tag. Mix half a teaspoon of non-detergent soap and one tablespoon baking soda in a gallon of water. doi: 10.1104/pp.15.00278, PubMed Abstract | CrossRef Full Text | Google Scholar, Apel, K., and Hirt, H. (2004). Mean values are indicated by . Median values are indicated by the middle line. For callose deposition visualization, rubber tree or N. benthamiana leaves were washed with double distilled water (ddH2O) and soaked in 95% ethanol, followed by boiling in water to clear the chlorophyll. The resulting proteins were then analyzed by western blot using anti-GFP, anti-RFP, anti--actin, and anti-Rubisco antibodies. Additionally, bacterial pathogens also secrete numerous effectors by the type III secretion system to target host plant plastids, including chloroplasts.
Similar to the Avr1b signal peptide, the EqCSEP01276 signal peptide, but not the empty vector, allowed YTK12 to grow on the YPRAAA medium (Figure 4). It is an indication that there is a problem that you need to address immediately. 15:e1007620. It is identical to white spots on your leaves. The most common pests are mealy bugs and spider mites. Plant Cell Physiol. If aphids have infected your rubber plant, you can squish them with your fingers or simply prune the infested areas. WM and XL planned and designed the research. Individual values obtained from nine replicates are indicated by dots. We found that callose deposition in N. benthamiana was induced by the infiltration of 100 and 150 M ABA into the leaves (Supplementary Figures 3A,B). You can put your plant near your windows, but be sure that you are filtering the sunlight. Powdery mildew infects a wide range of agricultural, economic, and ornamental plants, causing significant losses. You can also use baby wipes or cotton swabs with alcohol to wipe the dust. Curr. As the leaves of rubber plants are dark green in color, you may think that it is easy to spot any white spots. doi: 10.21769/bioprotoc.1257, Liyanage, K. K., Khan, S., Brooks, S., Mortimer, P. E., Karunarathna, S. C., Xu, J., et al. Asterisks indicate significant differences (P < 0.01). Plants suffer further damage due to dehydration when the temperature warms up. It has been reported that the murine p53 protein domain forms a complex with the 40 large T-antigen (Tan et al., 1986). J. Proteome Res. Individual values (n = 9) are indicated by dots.