猪粪—秸秆堆肥提取液促进植物生长和抑制土传病原菌效果及其作用机制
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摘要
堆肥提取液作为一种天然的液体肥料,由于其生防和肥效双重作用而具有十分广阔的应用前景。已经有很多文献报道堆肥提取液生防各种植物病害的效果,但堆肥提取液的生防效果很不稳定；另一方面除生防作用外,由于堆肥提取液中含有大量的有机和无机营养物质而使其还具有较好的促生效果。通常情况下,采用好气提取和厌气提取作为主要的提取方法,但是目前堆肥提取液在其提取过程中是否需要通气仍然存在争议。因此,本文以猪粪和水稻秸秆所生产的堆肥为原料,研究了采用直接提取、好气提取和厌气提取三种工艺提取的堆肥提取液的促生、生防土传病原真菌和番茄根结线虫(Meloidyne incognita)的效果及其作用机理。主要研究结果如下：
1、在好气条件下,最适提取工艺参数是提取比例为1：8,提取时间为36小时；而在厌气条件下,最适提取工艺参数是提取比例为1：8,提取时间为7天。
与直接提取比较,好气提取和厌气提取过程促进了猪粪-秸秆堆肥提取液中养分元素和有机高分子物质的累积。厌气(7天)提取猪粪-秸秆堆肥提取液(NAFEC)中全氮(TN)和铵态氮(NH4+)含量最高,比直接提取猪粪-秸秆堆肥提取液(DEC)、好气(36小时)提取猪粪-秸秆堆肥提取液(AFEC,通气量为33L·min-1)分别增加了16.4%和67.5%、36.0%和30.0%。DEC中全磷(TP)含量显著低于AFEC和NAFEC,分别降低了12.2%和20.6%。AFEC中全钾(TK)含量比DEC和NAFEC分别减少了4.5%和3.6%。不同提取工艺猪粪-秸秆堆肥提取液主要含有草酸和乙酸两种低分子量有机酸,AFEC中草酸和乙酸含量最高。
三种不同提取工艺猪粪-秸秆堆肥提取液中可培养微生物均以细菌为主。与DEC相比,NAFEC细菌丰度指数、多样性指数和稳定性指数均减少,而DEC、AFEC和NAFEC细菌均度指数则相当。
2、不同通气量显著影响猪粪-秸秆堆肥提取液的理化性质。AFEC-1(好气条件下通气量为11L·min-1提取的猪粪-秸秆堆肥提取液)和AFEC具有较高的腐殖化和芳构化水平以及分子结构中较低比例的羧基碳。
3.盆栽试验结果表明,不同提取工艺猪粪-秸秆堆肥提取液都能在一定程度上促进黄瓜生长代谢。AFEC处理黄瓜植株地上部干重最高,分别比DEC、NAFEC和1/2霍格兰-阿农营养液(CK)处理增加了34.5%、56.3%和63.9%,且处理间差异达到显著水平。AFEC处理黄瓜叶片碳氮代谢能力最高。
堆肥提取液的促生效果与其提取工艺有关。不同通气量显著影响猪粪-秸秆堆肥提取液的促生效果。以猪粪-秸秆堆肥为原料,采用好气提取工艺且通气量达到11L·min-1时制作的堆肥提取液就能获得较好的促生效果。
4、不同提取工艺猪粪-秸秆堆肥提取液中所含的乙酸和草酸对黄瓜的促生效果没有影响。好气提取猪粪-秸秆堆肥提取液中腐殖酸(H-AFEC)处理黄瓜地上部和根系氮素累积量最高,比直接提取猪粪-秸秆堆肥提取液中腐殖酸(H-DEC)、厌气提取猪粪-秸秆堆肥提取液中腐殖酸(H-NAFEC)、CK分别增加了9.1%和34.5%、21.8%和20.9%、19.2%和41.1%,H-AFEC与其他处理间差异达到显著水平。
5、不同提取工艺猪粪-秸秆堆肥提取液中腐殖酸均表现出类生长素的活性,H-DEC富含多糖碳具有最高的类生长素活性,H-NAFEC富含酚基碳具有最低的类生长素活性。H-NAFEC具有较高的腐熟度和芳构化水平。因此,较高的腐熟度和芳构化水平及酚基碳比例可能是导致H-NAFEC具有较低类生长素活性的主要原因。本试验条件下,由于三种提取工艺猪粪-秸秆堆肥提取液中未发现具有促生功能的有益微生物,因此,不同腐殖酸的浓度和类生长素活性是导致DEC、AFEC和NAFEC具有不同促生效果的主要原因。
6.除黄瓜立枯病病原菌外,猪粪-秸秆堆肥提取液均能显著地抑制各种植物病原真菌的生长。不同通气量的猪粪-秸秆堆肥提取液对黄瓜枯萎病病原真菌、黄瓜立枯病病原真菌和西瓜蔓枯病病原真菌的抑菌效果没有差异。不论采用哪种提取工艺,细菌在猪粪-秸秆堆肥提取液的抑菌生防效果中起主导作用。不同提取工艺猪粪-秸秆堆肥提取液细菌结构功能多样性指标不影响其抑菌效果。表明猪粪-秸秆堆肥提取液生防植物病原真菌的效果受提取工艺和病原菌种类的影响。
不同提取工艺猪粪-秸秆堆肥提取液对番茄根系中线虫虫卵的抑制率达到33.7%-87.9%,而对土壤中线虫密度的抑制率达到31.3%-51.4%。总之,采用合适的提取工艺,猪粪-秸秆堆肥提取液都能获得较好的生防根结线虫效果,其中,厌气(7天)提取猪粪-秸秆堆肥提取液促生和生防效果相对最好。
Compost extracts as natural liquid fertilizers with both bio-control and growth promotion effects have been widely used. Many evidences demonstrated that the compost extracts could suppress certain plant bacterial or fungal diseases, but the suppression effect was unstable. Compost extracts with some organic and inorganic nutrients could also significantly promoted plant growth. Aerated and non-aerated methods are the two dominant approaches in the production of compost extracts, though there is a debate about the necessity of aeration during the production of compost extracts. Therefore, this work compared the different technologies of compost extraction such as direct extraction, aerated fermentation and non-aerated fermentation methods, and then evaluated the plant growth promotion, inhibition of soilborne disease and reduction of tomato root-knot nematodes (Meloidyne incognita) as affected by the application of compost extracts that were produced from pig manure and rice straw. The acting mechanism of the compost extracts was also investigated. The main results are as follows:
1. The optimum process parameters were1:8of extraction ratio,36hours of extraction under the aerated condition, while, those under non-aerated condition is1:8of extraction ratio and7days of extraction for the compost extracts.
Compared with the direct extraction of pig manure-straw compost extracts, aerated and non-aerated fermentation extracts were rich in nutrient accumulation especially for high molecular weight substances. Non-aerated fermentation extracts of pig manure-straw compost (7days)(NAFEC) had the highest content of total nitrogen (TN) and ammonium nitrogen (NH4+), with an increase of16.4%and67.5%over direct extracts of pig manure-straw compost (DEC) and36%and30%over aerated fermentation extracts of pig manure-straw compost (36hours)(AFEC, with33L·min-1aeration quantity), respectively. When compared with AFEC and NAFEC, the total phosphorus (TP) content of DEC were decreased by12.2%and20.6%, respectively. The total potassium (TK) content of AFEC was lowest of all, and was decreased by4.5%and3.6%comparing with DEC and NAFEC, respectively. Acetic acid and oxalic acid were the two main organic acids with low molecular weights in compost extracts from pig manure-straw composts with different extraction technology. The highest content of oxalic acid and acetic acid was obtained by AFEC.
All compost extracts were determined to be dominated by bacteria. When compared with DEC, the bacterial richness index, diversity index and stability index of NAFEC were decreased; however, DEC had the equivalent bacterial evenness index with AFEC and NAFEC.
2. The physico-chemical characteristics were evidently affected by different aeration quantity. Aerated fermentation extracts of pig manure-straw compost with11L·min-1aeration quantity (AFEC-1) and AFEC had the high humification, aromatic levels and the low content of carboxyl groups.
3. The results of pot experiment showed that all pig manure-straw compost extracts promoted the carbon and nitrogen metabolism in cucumber plants. The maximum shoot dry weights of cucumber plants were obtained with the application of AFEC, which were34.5%,56.3%and63.9%higher than those with the applications of DEC, NAFEC, half-Hoagland and Arnon nutrient solution (CK), respectively. Compared with DEC and NAFEC, AFEC obtained the maximum cucumber leaf carbon and nitrogen metabolism ability.
The plant growth promotion of compost extracts was affected by the extraction technology. The aeration quantity affected significantly the growth promotion of aerated compost extracts. When the aeration quantity was11L·min-1, the compost extracts from pig manure-straw compost with aerated extraction technology could have a better growth promotion to cucumber plants.
4. There was no evident correlation between the contents of such organic acids as acetic acid and oxalic acid with low molecular weights and cucumber growth promotion by compost extracts from pig manure-straw compost with different extraction technologies. Humic-like substances extracted from AFEC (H-AFEC) obtained the maximum N accumulation content of shoot and root, with an increase of9.1%and34.5%over humic-like substances extracted from DEC (H-DEC),21.8%and20.9%over humic-like substances extracted from NAFEC (H-NAFEC) and19.2%and41.1%over CK. The difference between H-AFEC and other treatments were significant.
5. The H-DEC, H-AFEC and H-NAFEC all showed the positive auxin-like activity. The H-DEC was rich in peptidic and carbohydratic groups and showed the most auxin-like activity. The H-NAFEC was rich in phenolic groups and showed the least auxin-like activity. The H-AFEC showed a higher degree of aromatic condensation and humification. Thus, negative auxin-like activity of H-NAFEC could be attributed to the high degree of aromatic condensation and humification, and the low content of phenolic groups. In this experiment, there was no plant growth-promoting microorganism, therefore, the different growth promotion effects on cucumber plants with the application of DEC, AFEC and NAFEC might be caused by the different content of humic substances and distinguishing auxin-like activity.
6. Except for Rhizoctonia solania AG4, all compost extracts from pig manure-straw compost were remarkably antagonistic to the soilborne pathogenic fungi. Under different aerated conditions, there was no difference of the antagonistic effect of compost extracts against Fusarium oxysporum f.sp. cucumerinum, Didymella bryoniae and Rhizotctonia solania AG4. Microbial populations, especially for bacteria, were considered the most significant factor contributing to disease suppression of compost extracts. But, bacterial community diversity indexes of pig manure-straw compost extracts did not influence the inhibition effects on soilborne pathogen. It was shown that the inhibition effects of compost extracts on soilborne pathogenic fungi were not consistent, and were affected by the extraction technology and the plant pathogenic fungi variety.
All compost extracts could reduce tomato root nematode egg mass and density of soil nematodes more or less. The reduction ratio of root egg mass and density of soil nematodes were33.7%-87.9%and31.3%-51.4%, with the application of DEC, AFEC and NAFEC. In conclusion, the compost extracts from pig manure-straw compost could obtain the better reduction of root-knot nematodes, when extracted with reasonable extraction technology, the best growth promotion and bio-control effects could be obtained from the extracts with NAFEC technology.
1、在好气条件下,最适提取工艺参数是提取比例为1：8,提取时间为36小时；而在厌气条件下,最适提取工艺参数是提取比例为1：8,提取时间为7天。
与直接提取比较,好气提取和厌气提取过程促进了猪粪-秸秆堆肥提取液中养分元素和有机高分子物质的累积。厌气(7天)提取猪粪-秸秆堆肥提取液(NAFEC)中全氮(TN)和铵态氮(NH4+)含量最高,比直接提取猪粪-秸秆堆肥提取液(DEC)、好气(36小时)提取猪粪-秸秆堆肥提取液(AFEC,通气量为33L·min-1)分别增加了16.4%和67.5%、36.0%和30.0%。DEC中全磷(TP)含量显著低于AFEC和NAFEC,分别降低了12.2%和20.6%。AFEC中全钾(TK)含量比DEC和NAFEC分别减少了4.5%和3.6%。不同提取工艺猪粪-秸秆堆肥提取液主要含有草酸和乙酸两种低分子量有机酸,AFEC中草酸和乙酸含量最高。
三种不同提取工艺猪粪-秸秆堆肥提取液中可培养微生物均以细菌为主。与DEC相比,NAFEC细菌丰度指数、多样性指数和稳定性指数均减少,而DEC、AFEC和NAFEC细菌均度指数则相当。
2、不同通气量显著影响猪粪-秸秆堆肥提取液的理化性质。AFEC-1(好气条件下通气量为11L·min-1提取的猪粪-秸秆堆肥提取液)和AFEC具有较高的腐殖化和芳构化水平以及分子结构中较低比例的羧基碳。
3.盆栽试验结果表明,不同提取工艺猪粪-秸秆堆肥提取液都能在一定程度上促进黄瓜生长代谢。AFEC处理黄瓜植株地上部干重最高,分别比DEC、NAFEC和1/2霍格兰-阿农营养液(CK)处理增加了34.5%、56.3%和63.9%,且处理间差异达到显著水平。AFEC处理黄瓜叶片碳氮代谢能力最高。
堆肥提取液的促生效果与其提取工艺有关。不同通气量显著影响猪粪-秸秆堆肥提取液的促生效果。以猪粪-秸秆堆肥为原料,采用好气提取工艺且通气量达到11L·min-1时制作的堆肥提取液就能获得较好的促生效果。
4、不同提取工艺猪粪-秸秆堆肥提取液中所含的乙酸和草酸对黄瓜的促生效果没有影响。好气提取猪粪-秸秆堆肥提取液中腐殖酸(H-AFEC)处理黄瓜地上部和根系氮素累积量最高,比直接提取猪粪-秸秆堆肥提取液中腐殖酸(H-DEC)、厌气提取猪粪-秸秆堆肥提取液中腐殖酸(H-NAFEC)、CK分别增加了9.1%和34.5%、21.8%和20.9%、19.2%和41.1%,H-AFEC与其他处理间差异达到显著水平。
5、不同提取工艺猪粪-秸秆堆肥提取液中腐殖酸均表现出类生长素的活性,H-DEC富含多糖碳具有最高的类生长素活性,H-NAFEC富含酚基碳具有最低的类生长素活性。H-NAFEC具有较高的腐熟度和芳构化水平。因此,较高的腐熟度和芳构化水平及酚基碳比例可能是导致H-NAFEC具有较低类生长素活性的主要原因。本试验条件下,由于三种提取工艺猪粪-秸秆堆肥提取液中未发现具有促生功能的有益微生物,因此,不同腐殖酸的浓度和类生长素活性是导致DEC、AFEC和NAFEC具有不同促生效果的主要原因。
6.除黄瓜立枯病病原菌外,猪粪-秸秆堆肥提取液均能显著地抑制各种植物病原真菌的生长。不同通气量的猪粪-秸秆堆肥提取液对黄瓜枯萎病病原真菌、黄瓜立枯病病原真菌和西瓜蔓枯病病原真菌的抑菌效果没有差异。不论采用哪种提取工艺,细菌在猪粪-秸秆堆肥提取液的抑菌生防效果中起主导作用。不同提取工艺猪粪-秸秆堆肥提取液细菌结构功能多样性指标不影响其抑菌效果。表明猪粪-秸秆堆肥提取液生防植物病原真菌的效果受提取工艺和病原菌种类的影响。
不同提取工艺猪粪-秸秆堆肥提取液对番茄根系中线虫虫卵的抑制率达到33.7%-87.9%,而对土壤中线虫密度的抑制率达到31.3%-51.4%。总之,采用合适的提取工艺,猪粪-秸秆堆肥提取液都能获得较好的生防根结线虫效果,其中,厌气(7天)提取猪粪-秸秆堆肥提取液促生和生防效果相对最好。
Compost extracts as natural liquid fertilizers with both bio-control and growth promotion effects have been widely used. Many evidences demonstrated that the compost extracts could suppress certain plant bacterial or fungal diseases, but the suppression effect was unstable. Compost extracts with some organic and inorganic nutrients could also significantly promoted plant growth. Aerated and non-aerated methods are the two dominant approaches in the production of compost extracts, though there is a debate about the necessity of aeration during the production of compost extracts. Therefore, this work compared the different technologies of compost extraction such as direct extraction, aerated fermentation and non-aerated fermentation methods, and then evaluated the plant growth promotion, inhibition of soilborne disease and reduction of tomato root-knot nematodes (Meloidyne incognita) as affected by the application of compost extracts that were produced from pig manure and rice straw. The acting mechanism of the compost extracts was also investigated. The main results are as follows:
1. The optimum process parameters were1:8of extraction ratio,36hours of extraction under the aerated condition, while, those under non-aerated condition is1:8of extraction ratio and7days of extraction for the compost extracts.
Compared with the direct extraction of pig manure-straw compost extracts, aerated and non-aerated fermentation extracts were rich in nutrient accumulation especially for high molecular weight substances. Non-aerated fermentation extracts of pig manure-straw compost (7days)(NAFEC) had the highest content of total nitrogen (TN) and ammonium nitrogen (NH4+), with an increase of16.4%and67.5%over direct extracts of pig manure-straw compost (DEC) and36%and30%over aerated fermentation extracts of pig manure-straw compost (36hours)(AFEC, with33L·min-1aeration quantity), respectively. When compared with AFEC and NAFEC, the total phosphorus (TP) content of DEC were decreased by12.2%and20.6%, respectively. The total potassium (TK) content of AFEC was lowest of all, and was decreased by4.5%and3.6%comparing with DEC and NAFEC, respectively. Acetic acid and oxalic acid were the two main organic acids with low molecular weights in compost extracts from pig manure-straw composts with different extraction technology. The highest content of oxalic acid and acetic acid was obtained by AFEC.
All compost extracts were determined to be dominated by bacteria. When compared with DEC, the bacterial richness index, diversity index and stability index of NAFEC were decreased; however, DEC had the equivalent bacterial evenness index with AFEC and NAFEC.
2. The physico-chemical characteristics were evidently affected by different aeration quantity. Aerated fermentation extracts of pig manure-straw compost with11L·min-1aeration quantity (AFEC-1) and AFEC had the high humification, aromatic levels and the low content of carboxyl groups.
3. The results of pot experiment showed that all pig manure-straw compost extracts promoted the carbon and nitrogen metabolism in cucumber plants. The maximum shoot dry weights of cucumber plants were obtained with the application of AFEC, which were34.5%,56.3%and63.9%higher than those with the applications of DEC, NAFEC, half-Hoagland and Arnon nutrient solution (CK), respectively. Compared with DEC and NAFEC, AFEC obtained the maximum cucumber leaf carbon and nitrogen metabolism ability.
The plant growth promotion of compost extracts was affected by the extraction technology. The aeration quantity affected significantly the growth promotion of aerated compost extracts. When the aeration quantity was11L·min-1, the compost extracts from pig manure-straw compost with aerated extraction technology could have a better growth promotion to cucumber plants.
4. There was no evident correlation between the contents of such organic acids as acetic acid and oxalic acid with low molecular weights and cucumber growth promotion by compost extracts from pig manure-straw compost with different extraction technologies. Humic-like substances extracted from AFEC (H-AFEC) obtained the maximum N accumulation content of shoot and root, with an increase of9.1%and34.5%over humic-like substances extracted from DEC (H-DEC),21.8%and20.9%over humic-like substances extracted from NAFEC (H-NAFEC) and19.2%and41.1%over CK. The difference between H-AFEC and other treatments were significant.
5. The H-DEC, H-AFEC and H-NAFEC all showed the positive auxin-like activity. The H-DEC was rich in peptidic and carbohydratic groups and showed the most auxin-like activity. The H-NAFEC was rich in phenolic groups and showed the least auxin-like activity. The H-AFEC showed a higher degree of aromatic condensation and humification. Thus, negative auxin-like activity of H-NAFEC could be attributed to the high degree of aromatic condensation and humification, and the low content of phenolic groups. In this experiment, there was no plant growth-promoting microorganism, therefore, the different growth promotion effects on cucumber plants with the application of DEC, AFEC and NAFEC might be caused by the different content of humic substances and distinguishing auxin-like activity.
6. Except for Rhizoctonia solania AG4, all compost extracts from pig manure-straw compost were remarkably antagonistic to the soilborne pathogenic fungi. Under different aerated conditions, there was no difference of the antagonistic effect of compost extracts against Fusarium oxysporum f.sp. cucumerinum, Didymella bryoniae and Rhizotctonia solania AG4. Microbial populations, especially for bacteria, were considered the most significant factor contributing to disease suppression of compost extracts. But, bacterial community diversity indexes of pig manure-straw compost extracts did not influence the inhibition effects on soilborne pathogen. It was shown that the inhibition effects of compost extracts on soilborne pathogenic fungi were not consistent, and were affected by the extraction technology and the plant pathogenic fungi variety.
All compost extracts could reduce tomato root nematode egg mass and density of soil nematodes more or less. The reduction ratio of root egg mass and density of soil nematodes were33.7%-87.9%and31.3%-51.4%, with the application of DEC, AFEC and NAFEC. In conclusion, the compost extracts from pig manure-straw compost could obtain the better reduction of root-knot nematodes, when extracted with reasonable extraction technology, the best growth promotion and bio-control effects could be obtained from the extracts with NAFEC technology.
引文
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