CAJ | 학술논문

【目的】从生防菌株产生的拮抗作用酶及生防菌株对病原菌生长的影响2方面研究黄绿木霉(YGF9)和木贼镰刀菌(LX6F2)对杨树溃疡病病菌葡萄座腔菌的抑菌机制，为杨树溃疡病的生物防治提供参考。【方法】通过生防菌株在几丁质培养基、纤维素培养基、β－l，3－葡聚糖培养基和酪蛋白培养基上的反应和生长情况，判定生防菌株能否产生几丁质酶、纤维素酶、β－1，3－葡聚糖酶和蛋白酶；通过显微镜观察2种生防菌株对病原菌菌丝生长和孢子萌发的影响。【结果】菌株YGF9和LX6F2代谢过程中均能使β－1，3－葡聚糖培养基中的蓝色消失，说明2种菌株具有产β－1，3－葡聚糖酶的能力；2种菌株可以在酪蛋白培养基上形成水解圈，说明2种菌株具有产蛋白酶的能力；2种菌株在几丁质培养基和纤维素培养基上没有产生水解圈和透明圈，说明2种菌株可能没有产几丁质酶和纤维素酶的能力。菌株LX6F2的菌丝可以与病原菌菌丝平行生长、缠绕或穿插生长在病菌菌丝上，从而使得病原菌菌丝扭曲、变形、异常膨大；菌株YGF9的菌丝使得病原菌菌丝发生折叠、畸形、断裂，从而阻碍病原菌菌丝的生长；2种菌株的发酵产物均可导致杨树溃疡病菌葡萄座腔菌的孢子皱缩畸形、孢子内部结构发生变化，从而使病原菌孢子不能正常萌发；抑菌物质使病原菌芽管变短、扭曲畸形，从而芽管上长出的菌丝不能向前伸展，使孢子失去侵袭能力；生防菌株对孢子萌发的抑制率可达94%以上。【结论】2种生防菌株可通过自身产生的拮抗作用酶和影响病原菌菌丝生长和孢子萌发来达到对葡萄座腔菌的抑制作用。本试验中未发现2种生防菌株产生几丁质酶和纤维素酶，在后续研究中需用不同的试验方法进行验证。此外，本文仅对生防菌株产生拮抗作用酶进行了定性研究，没有计算其产酶量，若今后需要分离提取这些拮抗作用酶，则需要进一步对生防菌株的产酶量进行定量分析。【目的】從生防菌株產生的拮抗作用酶及生防菌株對病原菌生長的影響2方麵研究黃綠木黴(YGF9)和木賊鐮刀菌(LX6F2)對楊樹潰瘍病病菌葡萄座腔菌的抑菌機製，為楊樹潰瘍病的生物防治提供參攷。【方法】通過生防菌株在幾丁質培養基、纖維素培養基、β－l，3－葡聚糖培養基和酪蛋白培養基上的反應和生長情況，判定生防菌株能否產生幾丁質酶、纖維素酶、β－1，3－葡聚糖酶和蛋白酶；通過顯微鏡觀察2種生防菌株對病原菌菌絲生長和孢子萌髮的影響。【結果】菌株YGF9和LX6F2代謝過程中均能使β－1，3－葡聚糖培養基中的藍色消失，說明2種菌株具有產β－1，3－葡聚糖酶的能力；2種菌株可以在酪蛋白培養基上形成水解圈，說明2種菌株具有產蛋白酶的能力；2種菌株在幾丁質培養基和纖維素培養基上沒有產生水解圈和透明圈，說明2種菌株可能沒有產幾丁質酶和纖維素酶的能力。菌株LX6F2的菌絲可以與病原菌菌絲平行生長、纏繞或穿插生長在病菌菌絲上，從而使得病原菌菌絲扭麯、變形、異常膨大；菌株YGF9的菌絲使得病原菌菌絲髮生摺疊、畸形、斷裂，從而阻礙病原菌菌絲的生長；2種菌株的髮酵產物均可導緻楊樹潰瘍病菌葡萄座腔菌的孢子皺縮畸形、孢子內部結構髮生變化，從而使病原菌孢子不能正常萌髮；抑菌物質使病原菌芽管變短、扭麯畸形，從而芽管上長齣的菌絲不能嚮前伸展，使孢子失去侵襲能力；生防菌株對孢子萌髮的抑製率可達94%以上。【結論】2種生防菌株可通過自身產生的拮抗作用酶和影響病原菌菌絲生長和孢子萌髮來達到對葡萄座腔菌的抑製作用。本試驗中未髮現2種生防菌株產生幾丁質酶和纖維素酶，在後續研究中需用不同的試驗方法進行驗證。此外，本文僅對生防菌株產生拮抗作用酶進行瞭定性研究，沒有計算其產酶量，若今後需要分離提取這些拮抗作用酶，則需要進一步對生防菌株的產酶量進行定量分析。
【목적】종생방균주산생적길항작용매급생방균주대병원균생장적영향2방면연구황록목매(YGF9)화목적렴도균(LX6F2)대양수궤양병병균포도좌강균적억균궤제，위양수궤양병적생물방치제공삼고。【방법】통과생방균주재궤정질배양기、섬유소배양기、β－l，3－포취당배양기화락단백배양기상적반응화생장정황，판정생방균주능부산생궤정질매、섬유소매、β－1，3－포취당매화단백매；통과현미경관찰2충생방균주대병원균균사생장화포자맹발적영향。【결과】균주YGF9화LX6F2대사과정중균능사β－1，3－포취당배양기중적람색소실，설명2충균주구유산β－1，3－포취당매적능력；2충균주가이재락단백배양기상형성수해권，설명2충균주구유산단백매적능력；2충균주재궤정질배양기화섬유소배양기상몰유산생수해권화투명권，설명2충균주가능몰유산궤정질매화섬유소매적능력。균주LX6F2적균사가이여병원균균사평행생장、전요혹천삽생장재병균균사상，종이사득병원균균사뉴곡、변형、이상팽대；균주YGF9적균사사득병원균균사발생절첩、기형、단렬，종이조애병원균균사적생장；2충균주적발효산물균가도치양수궤양병균포도좌강균적포자추축기형、포자내부결구발생변화，종이사병원균포자불능정상맹발；억균물질사병원균아관변단、뉴곡기형，종이아관상장출적균사불능향전신전，사포자실거침습능력；생방균주대포자맹발적억제솔가체94%이상。【결론】2충생방균주가통과자신산생적길항작용매화영향병원균균사생장화포자맹발래체도대포도좌강균적억제작용。본시험중미발현2충생방균주산생궤정질매화섬유소매，재후속연구중수용불동적시험방법진행험증。차외，본문부대생방균주산생길항작용매진행료정성연구，몰유계산기산매량，약금후수요분리제취저사길항작용매，칙수요진일보대생방균주적산매량진행정량분석。
Objective]Poplar canker is one of the major biological disaster of poplar plantations in China. Techniques of biological control to restrain poplar canker are sustainable and effective. Trichoderma aureoviride strain YGF9 and Fusarium equiseti strain LX6F2 are two biocontrol strains which were isolated from poplar tissue and soil,respectively. They may have the potential to prevent from and control poplar canker. In order to explore the mechanism of these strains in controlling disease, biocontrol efficacy of the strain YGF9 and the strain LX6F2 on poplar canker caused by Botryosphaeria dothidea were investigated with its antagonism enzyme and inhibition of pathogen’s growth. [Method]Production of chitinase,cellulose enzyme,β-1,3-glucanase and protease by the two strains was determined through the reaction and growth status of the two biocontrol strains grown on chitin medium,cellulose medium,β-1,3-glucan culture medium and casein medium. Effects of the two biocontrol strains on the pathogen hyphal growth and spore germination were observed through a microscope. [Result]The results showed that the strains YGF9 and LX6F2 could make β-1,3 glucanase medium’s blue color disappear in metabolic process,indicating that the two strains could produce β-1,3-dextranase. The two strains had the ability to form hydrolyzed circle on casein medium,suggesting that the two strains had the ability to produce protease; The hydrolyzed circle and transparent circle were not generated by the two strains in chitin medium and cellulose hydrolysis medium,inferring that the two strains were not able to produce chitinase and cellulase. The strain LX6F2 hypha grew parallelly to pathogen hyphae,winded or interspersed with pathogen hyphae,so as to make the pathogen hyphae distortion,deformation and enlargement abnormal. The strain YGF9 hypha made pathogen hyphae folding,deformity,fracture,thereby preventing pathogen hyphal growth. The fermentation products of the two biocontrol strains could lead to spores of poplar canker pathogen B. dothidea deformity,and change in the spore internal structure, thus the pathogen spores couldn’t germinated normally. The antifungous substances could make germ tubes shorter and twist deformity,so that germ tubes couldn’t stretch forward. The inhibition rate of the biocontrol strains to pathogenic spore germination could reach to more than 94%.[Conclusion]It was found that the two biocontrol strains could produce antagonism enzymes and inhibit pathogen’s growth,which can interpret why the strain YGF9 and strain LX6F2 could prevent from poplar canker. Chitinase and cellulase produced by the two biocontrol strains were not found in the present experiment,for which,the further study is required by using different experimental methods for validation. In addition, the present study only studied the ability of the two biocontrol strains producing antagonism enzyme in qualitative analysis, the amount of enzyme production,the separation and application of these antagonism enzymes,as well as quantitative analysis of the amount should be studied later.