灰葡萄孢代谢产物的除草活性及活性物质的分离纯化
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摘要
本研究以来自不同地区不同寄主上的11个灰葡萄孢菌株为试材，研究了其代谢产物对番茄和辣椒的致毒活性及对反枝苋和马唐的除草活性。结果发现灰葡萄孢代谢产物对供试植物种子萌发后根和芽的伸长以及幼苗的生长都有一定的抑制作用，但是不同菌株间差异明显。其中BC1和BC4菌株的抑制作用最强，BC3、BC11和BC5菌株次之，其他菌株的毒力较弱。结果还发现，灰葡萄孢代谢产物对供试植物种子根伸长抑制作用较强的菌株，对其芽以及幼苗的生长也有较强的抑制作用。另外，对番茄和辣椒致毒活性较强的菌株对反枝苋和马唐的除草活性也较强。
试验确定了菌株BC4代谢产物中除草活性物质产生的适宜条件：即改良Fries培养基和查氏-道氏培养基(Ⅰ)：培养时间是35d；pH值为6；培养温度为25℃；静止培养；培养基中添加一定比例的番茄、辣椒的浸出汁并未提高灰葡萄孢代谢产物的除草活性。
通过叶片浸渍法和割茎幼苗法对灰葡萄孢代谢产物进行除草谱试验，结果表明，灰葡萄孢代谢产物具有较广泛的杀草范围，对反枝苋、裂叶牵牛、田旋花、稗草、马唐、牛筋、马齿苋、狗尾草、黎叶片和幼苗都有较强的抑制作用，其中尤以反枝苋为重。
灰葡萄孢培养滤液经乙酸乙酯萃取、乙酸乙酯萃取相的浓缩液即粗毒素，粗毒素经3次柱层析分离出一个除草活性段，该活性馏分经高效液相色谱分离得到一峰形尖锐而对称的除草活性组分。
Eleven isolates of Botrytis cinerea collected from different regions and hosts were cultured in Czapek-Doxbiotion liquid media and their metabolites were used as crude toxin to test their toxicity on two crops, tomato and pepper, and two weeds, Amaranthus retroflexus and Digitaria sanguinalis. The metabolites generally showed certain toxicity on the root and bud growth of plant seeds and the seeding growth of plants tested, but different isolates exhibited different activities. Isolate BC1 and BC4 had the highest toxicity, isolate BC3,BC11 and BC5 took the second place, the other isolates had less activities. The isolates tested that had higher toxicity on root growth of plant seeds often had higher toxicity on bud growth of plant seeds and the seeding growth of plants. It had also shown that the isolates tested that had higher toxic activity on tomato and pepper also had higher herbicidal activity on Amaranthus retroflexus and Digitaria sanguinalis.
The fitting culture conditions for isolate BC4 producing more toxin were Improvement Fries and Czapek-Doxbiotion(I) culture medium , culture time for thirty-five days, pH for 6, culture temperature for 25 , still cultivation, and that the juice of tomato and pepper cannot improve herbicidal activity on Amaranthus retroflexus.
The metabolites of isolate BC4 inhibit the seeding growth and caused necrotic spots on leaves of many weeds according the result of bioassay. These weeds are Amaranthus retroflexus, Digitaria sanguinalis, Convolvulus arvensis, Pharbitis nil Choisy, Chenopodium album, Setaria viridis, Echinochloa crusgalli, Portulaca oleracea, and Eleusine indica. Amaranthus retroflexus was most susceptible in tested weed.
Culture filtrate of Botrytis cinerea was extracted with ethyl acetate, the ethyl
acetate extracts were condensed, Condensed liquid was called crude toxin, then crude toxin were fractionated three times by column chromatography(CC) as described in the material and methods. One herbicidal fraction was collected. The fraction was further purified by high performance liquid chromatography (HPLC). then a active peak taking on acute and symmetrical shape was collected.
试验确定了菌株BC4代谢产物中除草活性物质产生的适宜条件：即改良Fries培养基和查氏-道氏培养基(Ⅰ)：培养时间是35d；pH值为6；培养温度为25℃；静止培养；培养基中添加一定比例的番茄、辣椒的浸出汁并未提高灰葡萄孢代谢产物的除草活性。
通过叶片浸渍法和割茎幼苗法对灰葡萄孢代谢产物进行除草谱试验，结果表明，灰葡萄孢代谢产物具有较广泛的杀草范围，对反枝苋、裂叶牵牛、田旋花、稗草、马唐、牛筋、马齿苋、狗尾草、黎叶片和幼苗都有较强的抑制作用，其中尤以反枝苋为重。
灰葡萄孢培养滤液经乙酸乙酯萃取、乙酸乙酯萃取相的浓缩液即粗毒素，粗毒素经3次柱层析分离出一个除草活性段，该活性馏分经高效液相色谱分离得到一峰形尖锐而对称的除草活性组分。
Eleven isolates of Botrytis cinerea collected from different regions and hosts were cultured in Czapek-Doxbiotion liquid media and their metabolites were used as crude toxin to test their toxicity on two crops, tomato and pepper, and two weeds, Amaranthus retroflexus and Digitaria sanguinalis. The metabolites generally showed certain toxicity on the root and bud growth of plant seeds and the seeding growth of plants tested, but different isolates exhibited different activities. Isolate BC1 and BC4 had the highest toxicity, isolate BC3,BC11 and BC5 took the second place, the other isolates had less activities. The isolates tested that had higher toxicity on root growth of plant seeds often had higher toxicity on bud growth of plant seeds and the seeding growth of plants. It had also shown that the isolates tested that had higher toxic activity on tomato and pepper also had higher herbicidal activity on Amaranthus retroflexus and Digitaria sanguinalis.
The fitting culture conditions for isolate BC4 producing more toxin were Improvement Fries and Czapek-Doxbiotion(I) culture medium , culture time for thirty-five days, pH for 6, culture temperature for 25 , still cultivation, and that the juice of tomato and pepper cannot improve herbicidal activity on Amaranthus retroflexus.
The metabolites of isolate BC4 inhibit the seeding growth and caused necrotic spots on leaves of many weeds according the result of bioassay. These weeds are Amaranthus retroflexus, Digitaria sanguinalis, Convolvulus arvensis, Pharbitis nil Choisy, Chenopodium album, Setaria viridis, Echinochloa crusgalli, Portulaca oleracea, and Eleusine indica. Amaranthus retroflexus was most susceptible in tested weed.
Culture filtrate of Botrytis cinerea was extracted with ethyl acetate, the ethyl
acetate extracts were condensed, Condensed liquid was called crude toxin, then crude toxin were fractionated three times by column chromatography(CC) as described in the material and methods. One herbicidal fraction was collected. The fraction was further purified by high performance liquid chromatography (HPLC). then a active peak taking on acute and symmetrical shape was collected.
引文
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