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海洋细菌AiL3和Kc-38菌株防治芒果炭疽病研究
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摘要
为了解本实验室原筛选的对芒果炭疽病菌具有较强拮抗作用的解淀粉芽孢杆菌(Bacillus amyloliquefaciens) AiL3菌株对芒果炭疽病的防治作用,本文对AiL3菌株及其活性物质室内外防治芒果炭疽病的效果进行了测定。另外,本文还对另一株从红树植物体内分离筛选的、对芒果炭疽病菌等植物病原具有较强拮抗作用的海洋细菌Kc-38菌株的分类地位及其抗菌活性物质与防病作用等进行了初探。结果如下:
     AiL3菌株发酵液及其除菌发酵上清液(胞外分泌物)对田间芒果叶和果炭疽病防治测定结果表明,分别在芒果出芽期或幼果期每隔15d喷施一次,连续3次喷施菌株发酵液及其胞外分泌物,45d后它们对叶片炭疽病的防效分别为6.83%和15.87%,加0.1%的吐温-80并稀释10倍后其防效分别为18.58%和47.54%;对果实炭疽病的防效分别为28.96%和28.52%,加0.1%的吐温-80并稀释10倍后其防效分别为43.86%和49.70%;其中加0.1%吐温-80并稀释10倍后的菌株胞外分泌物的防效与施保克稀释500倍液的防效相当,不存在显著差异。
     用AiL3菌株发酵液及其除菌发酵上清液(胞外分泌物)浸泡处理对采后芒果保鲜防病测定结果表明,它们对处理后不加接炭疽病菌的采后芒果炭疽病9d后的防效分别为66.67%和65.55%,12d后的防效分别为28.26%和31.06%,其间存在显著差异。施保克的防效显著高于菌株发酵液及其除菌发酵上清液的防效。套袋的采后芒果在接种炭疽病菌后分别于间隔24h和48h后,再用AiL3菌株发酵液及其除菌发酵上清液(胞外分泌物)处理的防病结果表明,菌株发酵液处理6d、9d后的防效分别为46.31%、43.24%和17.19%、20.00%,胞外分泌物处理6d、9d后的防效分别为49.47%、29.73%和32.80%、13.89%,均显著高于同期施保克的防效。以上结果表明,AiL3菌株对芒果炭疽病具有的良好生防效果,具有一定的开发利用前景。
     Kc-38菌株是本实验室从红树植物秋茄Kandelia candel体内分离得到的内生细菌,对峙生长法测定发现该菌株对芒果炭疽病菌Colletotrichum gloeosporioides和枯萎病菌Fusarium oxysporum等多种植物病原菌具有较强的抑制作用。根据菌体形态、培养特性、生理生化反应及16S rDNA序列分析将该菌株初步鉴定为解淀粉芽孢杆菌(Bacillus amyloliquefaciens)。该菌株对芒果炭疽病具有良好的防效,用菌株发酵液及其胞外分泌物处理9d后,对采后芒果炭疽病的防效分别达到65.12%和62.79%。进一步研究表明,其分泌的胞外抗菌物质为一类对热稳定和抗紫外照射的蛋白类物质。
The Bacillus amyloliquefaciens strain AiL3 was early screened as a stong antagonistic bacteria against the Colletotrichum gloeosporioides. In order to study the efficacy of strain AiL3 on control of the pre-and post-harvest mango anthracnose disease caused by C. gloe- osporioides, the biocontorl efficacy of the strain and its cellular exudates were tested in this paper. In addition, the taxonomic position, antifungal substances and biocontrol efficacy of the marine bacterial strain Kc-38 which was isolated from mangroves and had strong inhibitory activity against C. gloeosporioides were also studied in the paper.
     The result of using the strain AiL3 and its cellular exudates to control mango anthracnose disease in the fields showed that: the bacterial suspension and its cellular exudates had 6.83% and 15.87% control effect on the mango foliar anthracnose disease at 45d respectively after spraying three times every 15ds during mango budding and fruiting stages, and the control efficacy of 10-fold diluent mixed with 0.1%Tween-80 were 18.58% and 47.54% respectively. The control efficacy of the bacterial suspension and its cellular exudates to the mango furit anthracnose disease were 28.96% and 28.52% respectively, and the control efficacy of 10-fold diluent mixed with 0.1%Tween-80 were 43.86% and 49.70% respectively. And the 10-fold diluent of the bacterial cellular exudates mixed with 0.1%Tween-80 had the same contorl efficacy as the 45% Prochloraz diluted 500 times.
     The result of using the strain AiL3 and its cellular exudates to control postharvest mango anthracnose disease showed that: soaking the mango fruits with the bacterial suspension and its cellular exudates for 10 min without inoculated the pathogen C. gloeosporioides, their contorl efficacy were 66.67% and 65.55% at 9d, and 28.26% and 31.06% at 12d respectively. While the 45% Prochloraz diluted 500 times had better contorl efficacy of 85.19% and 57.61% at 9d and 12d respectively than the bacterial suspension and its cellular exudates, and there were marked differences among them.
     Bagged postharvest mango fruits were inoculated the pathogen C. gloeosporioides and then soaked with the bacterial suspension and its cellular exudates for 10 min 24h and 48h after inoculation respectively. The result showed that: the control efficacy of the bacterial suspension were 46.31%, 43.24% and 17.19%、20.00% at 6d and 9d respectively, and the control efficacy of its cellular exudates were 49.47%, 29.73% and 32.80%, 13.89% at 6d and 9d respectively. Their control efficacy to post-harvest mango anthracnose disease were both better than 45% Prochloraz diluted 500 times which had the control efficacy of 36.85%, 27.03% and 17.31%, 9.88% at 6d and 9d respectively. The results showed that good biocontrol efficacy was achieved. It indicated that AiL3 was a good candidate strain to biological control the mango anthracnose disease, and had a good prospect of development and utilization.
     The endophytic bacterial strain Kc-38 isolated from the tissue of Kandelia candel had strong inhibitory activity against many plant pathogenic fungi, such as Colletotrichum gloeosporioides, Fusarium oxysporum, and so on. According to the morphological, physiological and biochemical characteristics, and 16S rDNA sequence analysis, strain Kc-38 was identified as Bacillus amyloliquefaciens. The strain Kc-38 had good control efficacy on mango anthracnose disease caused by C. gloeosporioides. When tested on postharvest mango fruits and treated with the bacterial suspension and its cellular exudates, the control efficacy on the 9th d reached 65.12% and 62.79%, respectively. Further studies showed that the antibiotic substances produced by the strain were some kind of antifungal proteins. The proteins were found to be UV-tolerant and thermostable.
引文
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