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Published on: August 14, 2018
Recombinant anti-HIV MAP30, a ribosome inactivating protein: against plant virus and bacteriophage
Nafiseh Amirzadeh1, Ali Moghadam2, Ali Niazi3
1Institute of Biotechnology, College of Agriculture, Shiraz University, Shiraz, Iran.
Abstract:
The ribosome inactivating proteins (RIPs) efficiently decrease the microbial infections in plants. Momordica charantia MAP30 is a type I RIP that has not been investigated against plant viruses or bacteriophages. To evaluate of these activities, the recombinant MAP30 (rMAP30) was produced in the hairy roots of Nicotiana tabacum. Inoculation of 3 μg of transgenic total protein or 0.6 μg of rMAP30 against 0.1 μg of TMV reduced the leaf necrotic spots to 78.23% and 82.72%, respectively. The treatment of 0.1 μg of CMV with rMAP30 (0.6 μg) showed the reduction in the leaf necrotic spots to 85.8%. While the infection was increased after rMAP30 dilution. In the time interval assays, the leaves were first inoculated with 1 μg of rMAP30 or 0.1 μg of purified TMV or CMV agent for 6 h, then virus or protein was applied in order. This led the spot reduction to 35.22% and 67% for TMV, and 38.61% and 55.31% for CMV, respectively. In both the pre- and co-treatments of 1:10 or 1:20 diluted bacteriophage with 15 μg of transgenic total protein, the number and diameter of the plaques were reduced. The results showed that the highest inhibitory effect was observed in the pre-treatment assay of bacteriophage with transgenic total protein for 24 h. The decrease in the growth of bacteriophage caused more growth pattern of Escherichia coli. The results confirm that rMAP30 shows antibacterial activity against Streptococcus aureus and E. coli, antifungal activity against Candida albicans, and antiviral activity against CMV and TMV. Moreover, rMAP30 exhibits anti-phage activity for the first time. According to our findings, rMAP30 might be a valuable preservative agent in foods and beverages in the food industry as well as an antiviral and antimicrobial mixture in agriculture.
Insights
Momordica charantia MAP30 (a ribosome inactivating protein) effectively combats plant viruses like TMV and CMV, and bacteriophages. This study demonstrates its novel anti-phage activity and potential as a broad-spectrum antimicrobial agent in agriculture and food preservation.
Area of Science:
- Plant Pathology
- Biochemistry
- Microbiology
Background:
- Ribosome-inactivating proteins (RIPs) are known to reduce microbial infections in plants.
- Momordica charantia MAP30 is a type I RIP with potential antimicrobial properties.
- The antiviral and anti-phage activities of MAP30 have not been previously investigated.
Purpose of the Study:
- To produce recombinant MAP30 (rMAP30) in Nicotiana tabacum hairy roots.
- To evaluate the efficacy of rMAP30 against plant viruses (TMV, CMV) and bacteriophages.
- To assess the antibacterial and antifungal activities of rMAP30.
Main Methods:
- Recombinant MAP30 (rMAP30) was produced in transgenic hairy roots of Nicotiana tabacum.
- Inoculation assays were performed using TMV, CMV, and bacteriophages with varying concentrations of rMAP30 and transgenic protein.
- Time-interval assays and pre-/co-treatment strategies were employed to determine inhibitory effects.
- Antibacterial and antifungal activities were tested against E. coli, S. aureus, and C. albicans.
Main Results:
- rMAP30 significantly reduced necrotic spots caused by TMV (up to 82.72%) and CMV (up to 85.8%).
- Pre-treatment with rMAP30 showed higher efficacy in reducing viral and phage infections.
- rMAP30 demonstrated significant anti-phage activity, reducing plaque number and diameter.
- The protein exhibited broad-spectrum antimicrobial activity against bacteria and fungi.
Conclusions:
- rMAP30 possesses potent antiviral activity against TMV and CMV.
- This study presents the first evidence of rMAP30's anti-phage activity.
- rMAP30 shows promise as a natural preservative agent in the food industry and as an antimicrobial agent in agriculture.

