西洋参连作障碍微生态机制及生防放线菌的抗病作用
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摘要
西洋参是名贵药用植物,有着极高的药用及经济价值,连作障碍是西洋参种植中亟待解决的“瓶颈”问题。本文重点研究了陕西留坝西洋参连作障碍的微生态机制及生防放线菌的抗病作用,主要结果如下:
1.西洋参病健株根区根表土壤微生物区系研究表明:①病株根区土壤中细菌、放线菌数量较健株分别降低63.9%、33.5%,真菌增加160.1%;病株根区土壤B/F和A/F值较健株分别降低86.1%和74.3%。②病株根表土中细菌、真菌数量较健株增加31.4%、29.8%,放线菌数量降低79%。③西洋参病株根区土中土壤速效N、P、K及有机质含量分别较健株提高2.8%、35.4%、69.7%及10.2%。
2.西洋参病健株根内微生物区系及对病根病原真菌的分离鉴定研究表明:①在根表消毒条件下,病株根内细菌、真菌数量分别较健株增加3182.3%、40.6 %,在病健株根内均未分离到放线菌。②病株根内瓜小不整球壳属(Plectosphaerella cucumerina)占根内真菌总数的80.0%;从健株根内仅分离到茄腐皮镰孢菌(Fusarium solani)。
3.西洋参病健株根区根表土壤优势真菌有6种,通过ITS序列测定方法鉴定出的4种真菌为瓜小不整球壳属(Plectosphaerella cucumerina)、茄腐皮镰孢菌(Fusarium solani)、尖孢镰刀菌(Fusarium oxysporum)和绳状青霉(Penicillium funiculosum),其中瓜小不整球壳属在病、健根表土中所占比例分别为84.3%、46.1%,茄腐皮镰孢菌所占比例分别为13.7%、35.3%。瓜小不整球壳属在病株根表土壤中数量较健株增加2523.8%,该菌可能与西洋参发病有关。
4.生防放线菌的抑制作用及对病原真菌防治机理研究表明:①10株供试放线菌对5株西洋参人参病原真菌均有抑制作用,其中Act24-1、Act24-2对人参根腐菌的抑菌圈直径分别为22.5、21.5mm,Act24-2、Act7、D141及Act1对西洋参恶疫霉的抑菌圈直径为21.5~24.0mm。②8株放线菌无菌发酵液对病原菌菌丝生长均有抑制作用。其中菌株Act24-2对恶疫霉、人参锈腐病菌及人参根腐菌的相对抑菌率分别为83.3%、76.4%及64.4%。③5株供试病原菌均能诱导供试放线菌合成几丁质酶和纤维素酶;当供试放线菌与病原菌菌丝接触后,放线菌通过分泌几丁质酶、纤维素酶等真菌细胞壁降解酶,使病原菌菌丝溶解,抑制病原菌侵染,该结果从酶解角度揭示了生防放线菌对特定病原真菌的接触抗菌机理。
5.留坝西洋参病株病原真菌分离、鉴定及其对西洋参人参根系的侵染研究表明:①9株疑似病原真菌对西洋参、人参根系均表现出不同程度的侵染作用:茄腐皮镰孢菌(Fusarium solani)对人参和西洋参均有较强的侵染作用;西洋参人参受同一疑似病原真菌侵染后表现出不同程度的侵染症状,如尖孢镰刀菌对人参根系侵染作用强,但对西洋参根系侵染作用较弱,而菌株茄腐皮镰孢菌则相反。②留坝西洋参病害可能是由Fusarium solani(茄腐皮镰孢菌)和Fusarium oxysporum(尖孢镰刀菌)引起的。
6.西洋参种植土壤经过6年轮作倒茬后再种西洋参时发病严重,幼苗死亡率达95%以上。该西洋参地存活健株、病株根区根表土壤微生物区系研究表明:①弱株根区土中真菌、放线菌数量分别较健株增加39.9%、24.8%,细菌减少16.1%;弱株株根表土中细菌、真菌和放线菌分别较健株株增加4735.5%、74.4 %和64.4%。②病重株根表土中真菌、细菌较病轻株增加9970.3 %、297.5%。③弱株根区土壤养分均低于健株,其中铵态N和有机质含量分别较长势好株降低49.9%,55.1%。
American Ginseng is one of the famous and presious medicinal plant, with extremely high medicinal and economic value, but the continuous cropping obstacles of panax quinquefolium is urgently to be solved by the "bottleneck" problems. This paper maily studies the ecology mechanism of the continuous cropping obstacles on Panax quinquefoium in Liuba county of shaanxi and the resistance function by bio-control actinomycetes. The results are as follows.
1. The research on the soil microbial ecology between the rhizophere and root surface of the diseased and healthy American Ginseng shows that:①The number of bacteria and actinomyces in the rizosphere of diseased plant decreased by 63.9% and 33.5%, while fungi increased by 160.1%, compared with healthy plant. The B/F and A/F value in the rizosphere soil of diseased plant decreased by 86.1%,74.3%, respectively ,comparing to the healthy. The number of bacteria, fungi in the root surface soil of diseased plant increased by 31.4% and 29.8%, while actinomycetes decreased 79%.②The quantity of soil available nutrition (N, P, N) and organic matter in the rizophere soil of disesaed plant increased by 2.8%, 35.4%, 69.7% and 10.2%, respectively, compared with the healthy.
2. The study on the microbial ecology between the roots of the diseased and healthy American Ginseng and the separation of pathogenic fungi from the diseased root shows that:①The quantity of bacteria and fungi in the root of diseased plant increased 3182.3% and 40.6%, comparing to the healthy one; Actinomyces was not isolated from the diseased and healthy plant.②The percentage of Plectosphaerella cucumerina in the total root fungi is 80%. Fusarium solani was only isolated from the healthy American Ginseng.
3. There were 6 kinds of predominant fungi in the rhizosphere soil and root surface between the diseased and healthy American Ginseng. 4 kinds of fungi, identified by the ITS sequence analysis methods, were Plectosphaerella cucumerina, Fusarium solani, Fusarium oxysporum, Penicillium funiculosum. The proportion of Plectosphaerella cucumerina both in the soil surface of diseased and healthy plant were 84.3%, 46.1%, respectively. Similarly, Fusarium solani were 13.7%, 35.3%. Plectosphaerella cucumerina , the quantities of which in the root surface of diseased plant increased 2523.8%, comparing to the healthy plant, was closely related with the illness of American Ginseng, probably.
4. The research on inhibition and mechanism of bio-control actinomycetes against pathogen shows that:①10 strains of antigonistic actinomyces had inhibitory effects on the growth of fungal pathogen of American Ginseng and Ginseng. The diameters of inhibitory zones of Act24-1 and Act24-2 against Neonectria radicicola were 22.5, 21.5mm, respectively; The diameters of inhibitory zones of Act24-2, Act7,D141 and Act1 against Phytophthora cactorum were distributed between 21.5 and 24.0mm.②The axenic fermentation extracts of 8 strains of actinomyces can inhibit the growth of fungal pathogen. The inhibitory rates of Axenic fermentation extracts of Act24-2 to the Phytophthora cactorum, Cylindrocarpon sp. and Neonectria radicicola were 83.3%, 76.4% and 64.4%, respectively. 5 strains of fungal pathogen of Ginseng and American Ginseng could induce selected actinomyces to produce chitinases and cellulase. Selected actinomyces, contacting with the mycelia of pathogen, can dissolve the hyphae of pathogen and inhibit the infection of pathogen by producing cell wall degrading enzyme, such as chitinases and cellulase. This reveals the mechanism of bio-control actinomycetes against specific pathogen fungi from the angle of zymolysing.
5. The research on isolation, identification of the fungal pathogen from diseased root of American Ginseng in Liuba and infection of American Ginseng and Ginseng by suspected pathogens shows that:①9 strains of suspected pathogen repersented different degree of infection to the root of American Ginseng and Ginseng. Fusarium solani has strong infection to both American Ginseng and Ginseng; The root of American Ginseng and Ginseng showed different degree of symptoms after infected by the same suspected pathogen, such as Fusarium oxysporum can infected Ginseng root seriously, but had weak infection to American Ginseng root, however, Fusarium solani play the oppsite role.②The disease of American Ginseng in Liuba county was caused by Fusarium solani and Fusarium oxysporum, probably.
6. Serious diseases occurred to American Ginseng after 6-years crop rotation of the soil, the mortality of American Ginseng seedling was over 95%. The study on soil microbial ecology between rhizosphere and root surface of healthy American Ginseng and root surface of diseased one shows that:①For the healthy plant, the number of fungi and actinomyces in the rhizospere soil of the inferiror increased 39.9%, 24.8%,respectively, while the bacteria decreased 16.1%. The amount of bacteria, fungi and actinomyces in the root surface soil of inferior plant increased 4735.5%, 74.4% and 64.4%, respectively, comparing to the superior plant.②For the diseased plant, the amount of fungi and bacteria in root surface soil of heavy infected plant increased 9970.3%, 297.5%, comparing to the light infedcted one.③the quantity of soil nutrition of the inferior plant were lower than the superior one, of which, the amount of available N and organic matter decreased 49.9%, 55.1%, respectively, compared with the superior plant.
1.西洋参病健株根区根表土壤微生物区系研究表明:①病株根区土壤中细菌、放线菌数量较健株分别降低63.9%、33.5%,真菌增加160.1%;病株根区土壤B/F和A/F值较健株分别降低86.1%和74.3%。②病株根表土中细菌、真菌数量较健株增加31.4%、29.8%,放线菌数量降低79%。③西洋参病株根区土中土壤速效N、P、K及有机质含量分别较健株提高2.8%、35.4%、69.7%及10.2%。
2.西洋参病健株根内微生物区系及对病根病原真菌的分离鉴定研究表明:①在根表消毒条件下,病株根内细菌、真菌数量分别较健株增加3182.3%、40.6 %,在病健株根内均未分离到放线菌。②病株根内瓜小不整球壳属(Plectosphaerella cucumerina)占根内真菌总数的80.0%;从健株根内仅分离到茄腐皮镰孢菌(Fusarium solani)。
3.西洋参病健株根区根表土壤优势真菌有6种,通过ITS序列测定方法鉴定出的4种真菌为瓜小不整球壳属(Plectosphaerella cucumerina)、茄腐皮镰孢菌(Fusarium solani)、尖孢镰刀菌(Fusarium oxysporum)和绳状青霉(Penicillium funiculosum),其中瓜小不整球壳属在病、健根表土中所占比例分别为84.3%、46.1%,茄腐皮镰孢菌所占比例分别为13.7%、35.3%。瓜小不整球壳属在病株根表土壤中数量较健株增加2523.8%,该菌可能与西洋参发病有关。
4.生防放线菌的抑制作用及对病原真菌防治机理研究表明:①10株供试放线菌对5株西洋参人参病原真菌均有抑制作用,其中Act24-1、Act24-2对人参根腐菌的抑菌圈直径分别为22.5、21.5mm,Act24-2、Act7、D141及Act1对西洋参恶疫霉的抑菌圈直径为21.5~24.0mm。②8株放线菌无菌发酵液对病原菌菌丝生长均有抑制作用。其中菌株Act24-2对恶疫霉、人参锈腐病菌及人参根腐菌的相对抑菌率分别为83.3%、76.4%及64.4%。③5株供试病原菌均能诱导供试放线菌合成几丁质酶和纤维素酶;当供试放线菌与病原菌菌丝接触后,放线菌通过分泌几丁质酶、纤维素酶等真菌细胞壁降解酶,使病原菌菌丝溶解,抑制病原菌侵染,该结果从酶解角度揭示了生防放线菌对特定病原真菌的接触抗菌机理。
5.留坝西洋参病株病原真菌分离、鉴定及其对西洋参人参根系的侵染研究表明:①9株疑似病原真菌对西洋参、人参根系均表现出不同程度的侵染作用:茄腐皮镰孢菌(Fusarium solani)对人参和西洋参均有较强的侵染作用;西洋参人参受同一疑似病原真菌侵染后表现出不同程度的侵染症状,如尖孢镰刀菌对人参根系侵染作用强,但对西洋参根系侵染作用较弱,而菌株茄腐皮镰孢菌则相反。②留坝西洋参病害可能是由Fusarium solani(茄腐皮镰孢菌)和Fusarium oxysporum(尖孢镰刀菌)引起的。
6.西洋参种植土壤经过6年轮作倒茬后再种西洋参时发病严重,幼苗死亡率达95%以上。该西洋参地存活健株、病株根区根表土壤微生物区系研究表明:①弱株根区土中真菌、放线菌数量分别较健株增加39.9%、24.8%,细菌减少16.1%;弱株株根表土中细菌、真菌和放线菌分别较健株株增加4735.5%、74.4 %和64.4%。②病重株根表土中真菌、细菌较病轻株增加9970.3 %、297.5%。③弱株根区土壤养分均低于健株,其中铵态N和有机质含量分别较长势好株降低49.9%,55.1%。
American Ginseng is one of the famous and presious medicinal plant, with extremely high medicinal and economic value, but the continuous cropping obstacles of panax quinquefolium is urgently to be solved by the "bottleneck" problems. This paper maily studies the ecology mechanism of the continuous cropping obstacles on Panax quinquefoium in Liuba county of shaanxi and the resistance function by bio-control actinomycetes. The results are as follows.
1. The research on the soil microbial ecology between the rhizophere and root surface of the diseased and healthy American Ginseng shows that:①The number of bacteria and actinomyces in the rizosphere of diseased plant decreased by 63.9% and 33.5%, while fungi increased by 160.1%, compared with healthy plant. The B/F and A/F value in the rizosphere soil of diseased plant decreased by 86.1%,74.3%, respectively ,comparing to the healthy. The number of bacteria, fungi in the root surface soil of diseased plant increased by 31.4% and 29.8%, while actinomycetes decreased 79%.②The quantity of soil available nutrition (N, P, N) and organic matter in the rizophere soil of disesaed plant increased by 2.8%, 35.4%, 69.7% and 10.2%, respectively, compared with the healthy.
2. The study on the microbial ecology between the roots of the diseased and healthy American Ginseng and the separation of pathogenic fungi from the diseased root shows that:①The quantity of bacteria and fungi in the root of diseased plant increased 3182.3% and 40.6%, comparing to the healthy one; Actinomyces was not isolated from the diseased and healthy plant.②The percentage of Plectosphaerella cucumerina in the total root fungi is 80%. Fusarium solani was only isolated from the healthy American Ginseng.
3. There were 6 kinds of predominant fungi in the rhizosphere soil and root surface between the diseased and healthy American Ginseng. 4 kinds of fungi, identified by the ITS sequence analysis methods, were Plectosphaerella cucumerina, Fusarium solani, Fusarium oxysporum, Penicillium funiculosum. The proportion of Plectosphaerella cucumerina both in the soil surface of diseased and healthy plant were 84.3%, 46.1%, respectively. Similarly, Fusarium solani were 13.7%, 35.3%. Plectosphaerella cucumerina , the quantities of which in the root surface of diseased plant increased 2523.8%, comparing to the healthy plant, was closely related with the illness of American Ginseng, probably.
4. The research on inhibition and mechanism of bio-control actinomycetes against pathogen shows that:①10 strains of antigonistic actinomyces had inhibitory effects on the growth of fungal pathogen of American Ginseng and Ginseng. The diameters of inhibitory zones of Act24-1 and Act24-2 against Neonectria radicicola were 22.5, 21.5mm, respectively; The diameters of inhibitory zones of Act24-2, Act7,D141 and Act1 against Phytophthora cactorum were distributed between 21.5 and 24.0mm.②The axenic fermentation extracts of 8 strains of actinomyces can inhibit the growth of fungal pathogen. The inhibitory rates of Axenic fermentation extracts of Act24-2 to the Phytophthora cactorum, Cylindrocarpon sp. and Neonectria radicicola were 83.3%, 76.4% and 64.4%, respectively. 5 strains of fungal pathogen of Ginseng and American Ginseng could induce selected actinomyces to produce chitinases and cellulase. Selected actinomyces, contacting with the mycelia of pathogen, can dissolve the hyphae of pathogen and inhibit the infection of pathogen by producing cell wall degrading enzyme, such as chitinases and cellulase. This reveals the mechanism of bio-control actinomycetes against specific pathogen fungi from the angle of zymolysing.
5. The research on isolation, identification of the fungal pathogen from diseased root of American Ginseng in Liuba and infection of American Ginseng and Ginseng by suspected pathogens shows that:①9 strains of suspected pathogen repersented different degree of infection to the root of American Ginseng and Ginseng. Fusarium solani has strong infection to both American Ginseng and Ginseng; The root of American Ginseng and Ginseng showed different degree of symptoms after infected by the same suspected pathogen, such as Fusarium oxysporum can infected Ginseng root seriously, but had weak infection to American Ginseng root, however, Fusarium solani play the oppsite role.②The disease of American Ginseng in Liuba county was caused by Fusarium solani and Fusarium oxysporum, probably.
6. Serious diseases occurred to American Ginseng after 6-years crop rotation of the soil, the mortality of American Ginseng seedling was over 95%. The study on soil microbial ecology between rhizosphere and root surface of healthy American Ginseng and root surface of diseased one shows that:①For the healthy plant, the number of fungi and actinomyces in the rhizospere soil of the inferiror increased 39.9%, 24.8%,respectively, while the bacteria decreased 16.1%. The amount of bacteria, fungi and actinomyces in the root surface soil of inferior plant increased 4735.5%, 74.4% and 64.4%, respectively, comparing to the superior plant.②For the diseased plant, the amount of fungi and bacteria in root surface soil of heavy infected plant increased 9970.3%, 297.5%, comparing to the light infedcted one.③the quantity of soil nutrition of the inferior plant were lower than the superior one, of which, the amount of available N and organic matter decreased 49.9%, 55.1%, respectively, compared with the superior plant.
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