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重组抗 HIV MAP30,核糖体失活蛋白:抗植物病毒和噬菌体。

Recombinant anti-HIV MAP30, a ribosome inactivating protein: against plant virus and bacteriophage.

机构信息

Institute of Biotechnology, College of Agriculture, Shiraz University, Shiraz, Iran.

Center of Plant Virology Research, College of Agriculture, Shiraz University, Shiraz, Iran.

出版信息

Sci Rep. 2023 Feb 6;13(1):2091. doi: 10.1038/s41598-023-29365-7.

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.

摘要

核糖体失活蛋白(RIPs)可有效降低植物中的微生物感染。苦瓜 MAP30 是一种 I 型 RIP,尚未针对植物病毒或噬菌体进行研究。为了评估这些活性,在烟草毛状根中生产了重组 MAP30(rMAP30)。接种 3 μg 转基因总蛋白或 0.6 μg rMAP30 对 0.1 μg TMV 可分别将叶片坏死斑减少到 78.23%和 82.72%。用 rMAP30(0.6 μg)处理 0.1 μg CMV 可使叶片坏死斑减少到 85.8%。而 rMAP30 稀释后感染增加。在时间间隔测定中,首先将 1 μg rMAP30 或 0.1 μg 纯化 TMV 或 CMV 制剂接种叶片 6 h,然后依次接种病毒或蛋白。这导致 TMV 的斑点减少率分别为 35.22%和 67%,CMV 的斑点减少率分别为 38.61%和 55.31%。在 1:10 或 1:20 稀释的噬菌体与 15 μg 转基因总蛋白的预和共处理中,噬菌斑的数量和直径均减少。结果表明,在噬菌体与转基因总蛋白预处理 24 h 时观察到最高的抑制效果。噬菌体生长的减少导致大肠杆菌生长模式发生更多变化。结果证实 rMAP30 对金黄色葡萄球菌和大肠杆菌具有抗菌活性,对白色念珠菌具有抗真菌活性,对 CMV 和 TMV 具有抗病毒活性。此外,rMAP30 首次表现出抗噬菌体活性。根据我们的发现,rMAP30 可能是食品和饮料行业中一种有价值的防腐剂,也是农业中抗病毒和抗菌混合物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a323/9902390/f3d3a444e29c/41598_2023_29365_Fig1_HTML.jpg

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