Inhibition of Avicennia Marina antimicrobial protein against banana anthrax
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摘要: 本文以红树植物白骨壤叶为材料,以硫酸铵分级沉淀分离不同的抗菌蛋白,采用抑制菌丝生长速率和孢子萌发的方法测定白骨壤抗菌蛋白对香蕉炭疽病菌的抗菌活性,显微观察分析抗菌作用方式机理,进行白骨壤抗菌蛋白对感病香蕉的贮藏防病实验。结果表明:35%、75%、85%硫酸铵沉淀分离的抗菌蛋白（抗菌蛋白Ⅰ、Ⅱ、Ⅲ）具有明显抗菌活性,其抑制菌丝生长的半效应浓度(EC50)分别为:7.93±0.29、6.80±0.28、16.21±0.13μg/m L。100μg/m L的抗菌蛋白（Ⅰ、Ⅱ、Ⅲ）溶液对香蕉炭疽分生孢子的萌发抑制率分别达到95.5%±0.01%、97.5%±0.03%、92.5%±0.01%。显微观察发现三种抗菌蛋白处理的菌丝分别出现了扭曲、断裂、畸形以及原生质外渗现象。经抗菌蛋白处理后的采后感病香蕉果实具有明显降低发病率的效果。抗菌蛋白Ⅰ、Ⅱ、Ⅲ对香蕉炭疽病的防治效果分别达到74.48%±0.68%、77.24%±0.53%、75.87%±0.24%。Abstract: In this paper,the leaves of Avicennia Marina were selected as materials. The different antimicrobial proteins were isolated by ammonium sulphate precipitation on different saturation ranges. The inhibition ratio of antimicrobial proteins against banana anthrax was evaluated by mycelial growth rate and spore germination.Storage experiments of postharvest banana fruits treated by the antimicrobial proteins were carried out separately.The results showed that,the three type of antimicrobial proteins separated by 35%,75%,85% ammonium sulfate precipitation( antimicrobial protein Ⅰ,Ⅱ,Ⅲ) showed obvious antifungal activity,which inhibited mycelial growth and the EC50 value was 7.93 ± 0.29,16.21 ± 0.28,6.80 ± 0.13 μg / m L,respectively. The inhibition ratio of 100 μg / m L antimicrobial protein( Ⅰ,Ⅱ,Ⅲ) against spore germination was 95.5% ± 0.01%,97.5% ± 0.01% and 92.5% ± 0.01%respectively.By microexamination it was found that the three type of antimicrobial proteins could induce deformed growth of hyphae of banana anthrax,including twist of hyphae,necrosis of the hyphal wall and leakage of the cytoplasma.Application of the antimicrobial protein lead to a significant decrease in the incidence of banana anthrax during storage of postharvest banana fruits. The control efficiency of antimicrobial protein( Ⅰ,Ⅱ,Ⅲ) on banana anthracnose was 74.48% ± 0.68%,77.24% ± 0.53% and 75.87% ± 0.24%,respectively.
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[1] .李玉萍,方佳.中国香蕉产业现状与发展对策研究[J].中国农学通报,2008,24(8):443-447. [2] 李玉萍,方佳,梁伟红,等.国内外香蕉中农药残留限量标准的比较分析[J].热带作物学报,2006(4):113-118. [3] Jat B L,Sharma P,Gour H N.Production of Enzymes and Culture Filtrates by Colletotrichum gloeosporioides Penz.Causing Banana Fruit Rot[J].Proceedings of the National Academy of Sciences,India Section B:Biological Sciences,2013,83(2):177-180.
[4] Udayanga D,Manamgoda D S,Liu X,et al.What are the common anthracnose pathogens of tropical fruits?[J].Fungal Diversity,2013,61(1):165-179.
[5] 胡美姣,李敏,高兆银,等.香蕉炭疽病菌不同菌株的特性比较[J].热带作物学报,2007,28(2):87-91. [6] 周亚奎,陈旭玉,郑服丛.香蕉炭疽病生物防治研究进展[J].中国农学通报,2008,24(4):328-331. [7] Bautista-Rosales P U,Calderon-Santoyo M,Servín-Villegas R,et al.Biocontrol action mechanisms of Cryptococcus laurentii on Colletotrichum gloeosporioides of mango[J].Crop Protection,2014,65:194-201.
[8] Maqbool M,Ali A,Alderson P G,et al.Postharvest application of gum arabic and essential oils for controlling anthracnose and quality of banana and papaya during cold storage[J].Postharvest Biology and Technology,2011,62(1):71-76.
[9] 张娜,关文强,阎瑞香.芥末精油对芒果采后病原真菌抑制效果和保鲜效应的研究[J].食品工业科技,2011,32(3):349-353. [10] De los Santos-Villalobos S,Guzmán-Ortiz D A,GómezLim M A,et al.Potential use of Trichoderma asperellum(Samuels,Liechfeldt et Nirenberg)T8a as a biological control agent against anthracnose in mango(Mangifera indica L.)[J].Biological Control,2013,64(1):37-44.
[11] ivkovic'S,Stojanovic'S,Ivanovic',et al.Screening of antagonistic activity of microorganisms against Colletotrichum acutatum and Colletotrichum gloeosporioides[J].Archives of Biological Sciences,2010,62(3):611-623.
[12] Veronese P,Ruiz M T,Coca M A,et al.In defense against pathogens.Both plant sentinels and foot soldiers need to know the enemy[J].Plant Physiology,2003,131(4):1580-1590.
[13] Mc Manus A M,Nielsen K J,Marcus J P,et al.Mi AMP1,a novel protein from Macadamia integrifolia adopts a greek keyβ-barrel fold unique amongst plant antimicrobial proteins[J].Journal of Molecular Biology,1999,293(3):629-638.
[14] Regente M C,Giudici A M,Villalain J,et al.The cytotoxic properties of a plant lipid transfer protein involve membrane permeabilization of target cells[J].Letters in Applied Microbiology,2005,40(3):183-189.
[15] Kovalskaya N,Hammond R W.Expression and functional characterization of the plant antimicrobial snakin-1 and defensin recombinant proteins[J].Protein Expression and Purification,2009,63(1):12-17.
[16] Arazi T,Huang P L,Huang P L,et al.Production of antiviral and antitumor proteins MAP30 and GAP31 in cucurbits using the plant virus vector ZYMV-AGII[J].Biochemical and Biophysical Research Communications,2002,292(2):441-448.
[17] Che Y Z,Li Y R,Zou H S,et al.A novel antimicrobial protein for plant protection consisting of a Xanthomonas oryzae harpin and active domains of cecropin A and melittin[J].Microbial biotechnology,2011,4(6):777-793.
[18] Sun J,Xia Y,Li D,et al.Relationship between peptide structure and antimicrobial activity as studied by de novo designed peptides[J].Biochimica et Biophysica Acta(BBA)-Biomembranes,2014,1838(12):2985-2993.
[19] Seo J K,Lee M J,Jung H G,et al.Antimicrobial function of SHb AP,a novel hemoglobinβchain-related antimicrobial peptide,isolated from the liver of skipjack tuna,Katsuwonus pelamis[J].Fish&Shellfish Immunology,2014(37):173-183.
[20] 石莉.中国红树林的分布状况、生长环境及其环境适应性[J].海洋环境保护,2002(4):14-18. [21] 贾睿,郭跃伟,侯惠欣.中国红树林植物白骨壤化学成分的研究[J].中国天然药物,2004,2(1):16-19. [22] Kumar C,Karthik L,Bhaskara K.V.In vitro anti-candida activity of Calotropis gigantea against clinical isolates of Candida[J].Pharm Res 2010;3:539-542.
[23] 潘磊庆,朱娜,邵兴锋,等.丁香精油对樱桃番茄的保鲜作用[J].食品工业科技,2012,33(23):335-338. [24] 何红,蔡学清,陈玉森,等.辣椒内生枯草芽孢杆菌BS-2和BS-1防治香蕉炭疽病[J].福建农林大学学报(自然科学版),2002,31(4):442-444. [25] Grewal S,Bhagat M,Vakhlu J.Antimicrobial protein produced by pseudomonas aeruginosa JU-Ch 1,with a broad spectrum of antimicrobial activity[J].Biocatalysis and Agricultural Biotechnology,2014,3(4):332-337.
[26] Pelegrini P B,Noronha E F,Muniz M A R,et al.An antifungal peptide from passion fruit(Passiflora edulis)seeds with similarities to 2S albumin proteins[J].Biochimica et Biophysica Acta(BBA)-Proteins and Proteomics,2006,1764(6):1141-1146.
[27] Oard S V,Enright F M.Expression of the antimicrobial peptides in plants to control phytopathogenic bacteria and fungi[J].Plant Cell Rep,2006(25):561-572.
[28] Zasloff M.Antimicrobial peptides of multicellular organisms[J].nature,2002,415(24):389-395.
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