具杀线虫活性植物内生细菌的筛选和活性产物
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江苏省自然科学基金(BK2007222)


Screening endophytic bacteria against plant-parasitic nematodes
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Supported by the Natural Science Foundation of Jiangsu Province (BK2007222)

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    摘要:【目的】植物寄生线虫是危害植物的重要病原物,为了筛选到能在植物体内稳定定殖并且对植物寄生线虫具有较高杀线虫活性的植物内生细菌生防菌。【方法】以松材线虫为靶标,用直接触杀法进行筛选。对高活性菌株采用正交实验优化发酵条件,测定发酵液杀线虫活性的稳定性,并对菌株进行鉴定。【结果】从6种植物中分离筛选出13株对松材线虫具有较高杀线虫活性的植物内生细菌菌株,这些菌株的发酵上清液对松材线虫处理24h杀线虫率均达到了100%;其中BCM2、SZ5、CCM7和DP1这4个菌株的杀线活性较高,发酵上清液稀释3倍处理2

    Abstract:

    Abstract: [Objective] Plant-parasite nematode is one of the most important pathogens in plant. Our objective is to screen endophytic bacteria against plant-parasitic nematodes from plant. [Methods] Endophytic bacteria were isolated and screened by testing their metabolite against Bursaphelenchus xylophilus in vitro. Those strains inhibiting B. xylophilus were selected to culture in liquid medium and fermentation conditions were optimized by orthogonal test. The stability of the antinematode substances was evaluated by various. In addition, four strains were identified by 16SrDNA sequence analysis. [Results] In total 13 strains of endophytic bacteria secreting antinematode metabolite were isolated from 6 species of plant. The supernatant of the fermentation broth of these endophytic bacteria gave 100% mortality of nematodes after treated as the follows:1ml each was mixed with 0.2ml of the suspension of nematodes (2000 nematodes/ml) then incubated at 25℃ for 24 h, some of which could led to leakage or dissolution of nematodes. Among them, four strains, BCM2、SZ5、CCM7 and DP1, showed stronger activity than others. The supernatants diluted three times also gave not less than 95% mortality after 24 h treatment, and those from DP1 and SZ5 even gave 100% mortality. The fermentation conditions of the four strains were optimized and the antinematode activity grew up four times after optimization. The antinematode substances of these strains were found stable when treated with protease or heating or stored at 4℃ after 100 days, while instable when treated with acid or alkali. DP1 and CCM7 were identified to be Bacillus subtilis, while SZ5 and BCM2 to be Bacillus cereus. [Conclusion] Endophytic bacteria secreting antinematode metabolite were found in economic crops. The metabolite of some strains showed strong and stable antinematode activity. Our results indicate the real potential of biocontrol by endophytic bacteria.

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彭双,闫淑珍,陈双林. 具杀线虫活性植物内生细菌的筛选和活性产物. 微生物学报, 2011, 51(3): 368-376

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  • 收稿日期:2010-09-28
  • 最后修改日期:2010-12-10
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