Limits...
A novel antimicrobial protein for plant protection consisting of a Xanthomonas oryzae harpin and active domains of cecropin A and melittin.

Che YZ, Li YR, Zou HS, Zou LF, Zhang B, Chen GY - Microb Biotechnol (2011)

Bottom Line: The resulting chimeric protein maintained not only the HR-inducing property of the harpin, but also the antimicrobial activity of the cecropin A-melittin hybrid.Importantly, the protein acted as a potential pesticide by inducing disease resistance for viral, bacterial and fungal pathogens.This designed drug can be considered as a lead compound for use in plant protection, either for the development of new broad-spectrum pesticides or for expression in transgenic plants.

View Article: PubMed Central - PubMed

Affiliation: Department of Plant Pathology, Nanjing Agricultural University/Key Laboratory of Monitoring and Management for Plant Diseases and Insects, Ministry of Agriculture of China, Nanjing 210095, China.

Show MeSH

Related in: MedlinePlus

Antimicrobial activity of Hcm1 against bacteria and fungi on agar plates. 1. Kanamycin (10 µg ml−1) for bacteria and carbendazim (50 µg ml−1) for fungi were used as positive controls; 2. Hcm1 (5 µM); 3. Hcm1 (1 µM); 4. Hpa1 (5 µM). The sterile filter paper discs (5 mm diameter) were dipped in the above solutions, respectively, and then laid on NA plates where 100 µl of the respective bacteria (below) at approximately 1 × 106 cfu ml−1 had been spread previously. After 2 days incubation at 28°C, antimicrobial haloes around the discs were recorded. For antifungal tests, 5‐mm‐dia mycelial discs of the fungi listed below were placed on PDA plates, and then 10 µl of the above solutions were added to 5‐mm‐dia holes made around the mycelial discs by a hole punch. Antimicrobial haloes around the holes were recorded after 5 days at 28°C, depending on different fungal growth. Similar results were obtained from three replications. (A) E. coli BL21 (DE3); (B) R. solanacearum ZJ3721; (C) X. oryzae pv. oryzicola RS105; (D) P. syringae pv. tomato DC3000; (E) B. subtilis B168; (F) A. alternata TBA28; (G) M. oryzae Guy11; (H) F. graminearum ZF21; (I) T. cucumeris JS01.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3815413&req=5

f4: Antimicrobial activity of Hcm1 against bacteria and fungi on agar plates. 1. Kanamycin (10 µg ml−1) for bacteria and carbendazim (50 µg ml−1) for fungi were used as positive controls; 2. Hcm1 (5 µM); 3. Hcm1 (1 µM); 4. Hpa1 (5 µM). The sterile filter paper discs (5 mm diameter) were dipped in the above solutions, respectively, and then laid on NA plates where 100 µl of the respective bacteria (below) at approximately 1 × 106 cfu ml−1 had been spread previously. After 2 days incubation at 28°C, antimicrobial haloes around the discs were recorded. For antifungal tests, 5‐mm‐dia mycelial discs of the fungi listed below were placed on PDA plates, and then 10 µl of the above solutions were added to 5‐mm‐dia holes made around the mycelial discs by a hole punch. Antimicrobial haloes around the holes were recorded after 5 days at 28°C, depending on different fungal growth. Similar results were obtained from three replications. (A) E. coli BL21 (DE3); (B) R. solanacearum ZJ3721; (C) X. oryzae pv. oryzicola RS105; (D) P. syringae pv. tomato DC3000; (E) B. subtilis B168; (F) A. alternata TBA28; (G) M. oryzae Guy11; (H) F. graminearum ZF21; (I) T. cucumeris JS01.

Mentions: To verify our earlier results using the CFEPs (Fig. 2), that the antibacterial activity of Hcm1 is due to the addition of the cecropin A‐melittin hybrid to the Hpa1 backbone, we used purified Hcm1 to test for growth inhibition of E. coli on LB plates, using Hpa1 at 5 µM as a negative control and kanamycin (Km) at 10 µg ml−1 as a positive control. Indeed, the application of 0.5 µM of Hcm1 to a sterile paper disc caused an obvious inhibition halo against E. coli BL21 (DE3), while Hpa1 did not (Fig. 4A), confirming that the addition of the cecropin A‐melittin hybrid at the C‐terminus of Hpa1 confers antibacterial activity to Hcm1.


A novel antimicrobial protein for plant protection consisting of a Xanthomonas oryzae harpin and active domains of cecropin A and melittin.

Che YZ, Li YR, Zou HS, Zou LF, Zhang B, Chen GY - Microb Biotechnol (2011)

Antimicrobial activity of Hcm1 against bacteria and fungi on agar plates. 1. Kanamycin (10 µg ml−1) for bacteria and carbendazim (50 µg ml−1) for fungi were used as positive controls; 2. Hcm1 (5 µM); 3. Hcm1 (1 µM); 4. Hpa1 (5 µM). The sterile filter paper discs (5 mm diameter) were dipped in the above solutions, respectively, and then laid on NA plates where 100 µl of the respective bacteria (below) at approximately 1 × 106 cfu ml−1 had been spread previously. After 2 days incubation at 28°C, antimicrobial haloes around the discs were recorded. For antifungal tests, 5‐mm‐dia mycelial discs of the fungi listed below were placed on PDA plates, and then 10 µl of the above solutions were added to 5‐mm‐dia holes made around the mycelial discs by a hole punch. Antimicrobial haloes around the holes were recorded after 5 days at 28°C, depending on different fungal growth. Similar results were obtained from three replications. (A) E. coli BL21 (DE3); (B) R. solanacearum ZJ3721; (C) X. oryzae pv. oryzicola RS105; (D) P. syringae pv. tomato DC3000; (E) B. subtilis B168; (F) A. alternata TBA28; (G) M. oryzae Guy11; (H) F. graminearum ZF21; (I) T. cucumeris JS01.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3815413&req=5

f4: Antimicrobial activity of Hcm1 against bacteria and fungi on agar plates. 1. Kanamycin (10 µg ml−1) for bacteria and carbendazim (50 µg ml−1) for fungi were used as positive controls; 2. Hcm1 (5 µM); 3. Hcm1 (1 µM); 4. Hpa1 (5 µM). The sterile filter paper discs (5 mm diameter) were dipped in the above solutions, respectively, and then laid on NA plates where 100 µl of the respective bacteria (below) at approximately 1 × 106 cfu ml−1 had been spread previously. After 2 days incubation at 28°C, antimicrobial haloes around the discs were recorded. For antifungal tests, 5‐mm‐dia mycelial discs of the fungi listed below were placed on PDA plates, and then 10 µl of the above solutions were added to 5‐mm‐dia holes made around the mycelial discs by a hole punch. Antimicrobial haloes around the holes were recorded after 5 days at 28°C, depending on different fungal growth. Similar results were obtained from three replications. (A) E. coli BL21 (DE3); (B) R. solanacearum ZJ3721; (C) X. oryzae pv. oryzicola RS105; (D) P. syringae pv. tomato DC3000; (E) B. subtilis B168; (F) A. alternata TBA28; (G) M. oryzae Guy11; (H) F. graminearum ZF21; (I) T. cucumeris JS01.
Mentions: To verify our earlier results using the CFEPs (Fig. 2), that the antibacterial activity of Hcm1 is due to the addition of the cecropin A‐melittin hybrid to the Hpa1 backbone, we used purified Hcm1 to test for growth inhibition of E. coli on LB plates, using Hpa1 at 5 µM as a negative control and kanamycin (Km) at 10 µg ml−1 as a positive control. Indeed, the application of 0.5 µM of Hcm1 to a sterile paper disc caused an obvious inhibition halo against E. coli BL21 (DE3), while Hpa1 did not (Fig. 4A), confirming that the addition of the cecropin A‐melittin hybrid at the C‐terminus of Hpa1 confers antibacterial activity to Hcm1.

Bottom Line: The resulting chimeric protein maintained not only the HR-inducing property of the harpin, but also the antimicrobial activity of the cecropin A-melittin hybrid.Importantly, the protein acted as a potential pesticide by inducing disease resistance for viral, bacterial and fungal pathogens.This designed drug can be considered as a lead compound for use in plant protection, either for the development of new broad-spectrum pesticides or for expression in transgenic plants.

View Article: PubMed Central - PubMed

Affiliation: Department of Plant Pathology, Nanjing Agricultural University/Key Laboratory of Monitoring and Management for Plant Diseases and Insects, Ministry of Agriculture of China, Nanjing 210095, China.

Show MeSH
Related in: MedlinePlus