紫外线照射提高哈茨木霉和绿霉本地分离株的拮抗和抗杀菌剂耐受性

Abdullah Abdul Kareem Hassan
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The optimum exposure time to ultraviolet irradiation is 30 min, in which, the percentage of kill was 98-99% .After UV-irradiation treatment (Quartz lamps 30W at 240-280 nm, peak 254nm, distance from the source was 20 cm), antagonistic capability against R. solani and F. oxysporum was improved in mutants MT3,MT5,MS1 and MS6, in which antagonistic reaction score were 3.6,3.9,3.2 and 3.6 against F. oxysporum and 3.7,4,3.4 and 4 against R. solani compared to 3.2, 2.8, 2.6 and 2.8 against F. oxysporum and 2.7, 2.9,3.1 and 3 against R. solani in the wild types T3, T5, S1 and S6,respectively. The growth of T. harzianum isolates (estimated as colony diameter) was significantly increased in all T. harzianum mutants except MT3 and MD5 mutants, CMCase activity was increased from 13.2, 16.2, 10 and 15.8 U/ml / mg protein in the wild isolates T3, T5, S1 and S6 to 18.6, 20.3, 17.7 and 22.5 U/ml / mg protein in the mutants MT3, MT5, MS1 and MS6, respectively. 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引用次数: 1

摘要

在22株伊拉克哈茨木霉和绿僵菌分离株中,与哈茨木霉相关的T3、T5、D5、S1和S6对植物病原真菌尖孢镰刀菌和茄枯丝核菌表现出较高的拮抗活性。选择这些菌株,利用紫外照射提高其生物防治活性。紫外照射(石英灯30W, 240 ~ 280 nm,峰值254nm,距离源20 cm)后,突变体MT3、MT5、MS1和MS6对枯萎病菌和尖孢镰刀菌的拮抗能力得到提高,对枯萎病菌的拮抗反应得分分别为3.6、3.9、3.2和3.6,对枯萎病菌的拮抗反应得分分别为3.7、4、3.4和4,而对枯萎病菌的拮抗反应得分分别为3.2、2.8,野生型T3、T5、S1和S6对尖孢镰刀菌的抗性分别为2.6、2.8和2.7、2.9、3.1和3。除MT3和MD5突变体外,所有突变体的菌落直径均显著增加,CMCase活性分别从野生分离株T3、T5、S1和S6的13.2、16.2、10和15.8 U/ml / mg蛋白增加到MT3、MT5、MS1和MS6的18.6、20.3、17.7和22.5 U/ml / mg蛋白。这些突变体的几丁质酶活性分别为88.4、132.3、86.1和136.7 U/ml / mg蛋白,而野生的几丁质酶活性分别为82.7、93.2、74.5和103.2 U/ml / mg蛋白。对比野生型和突变体,在Topsin和Benomyl推荐田间用量的25%条件下,MT5的最大菌落直径为5.2 cm和4.8 cm,而野生型T5的最大菌落直径为3.3 cm和2.2 cm,其次是MS6,在这两种杀菌剂的相同浓度下,该突变体的最大菌落直径为4.8 cm和4.2 cm,而野生型S6的最大菌落直径为3.4 cm和2.5 cm。综上所示,紫外辐照培养的突变体MT5和MS6具有拮抗活性(在支寄生虫水平),生长速度快,CMCase和几丁质酶活性较高,在25%的Elsa、25%和50%的Diathen和Mizab推荐田间剂量以及所有浓度的Topsin和Benomyl下均能生长,对病原菌尖孢镰刀菌(F. oxysporum)和番茄枯萎菌(R. solani)具有完全抑制作用。
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Improvement of antagonism and fungicides tolerance in Trichoderma harzianum and T.viride local isolates by Ultra-Violet irradiation
Among 22 Iraqi Trichoderma harzianum  and T.viride isolates, T3,T5,D5,S1 and S6 related to T. harzianum  showed a higher antagonistic activity against the plant pathogenic fungi  Fusarium oxysporum and Rhizoctonia solani. These isolates were selected for improvement their biocontrol activity by usi ng uv-irradiation. The optimum exposure time to ultraviolet irradiation is 30 min, in which, the percentage of kill was 98-99% .After UV-irradiation treatment (Quartz lamps 30W at 240-280 nm, peak 254nm, distance from the source was 20 cm), antagonistic capability against R. solani and F. oxysporum was improved in mutants MT3,MT5,MS1 and MS6, in which antagonistic reaction score were 3.6,3.9,3.2 and 3.6 against F. oxysporum and 3.7,4,3.4 and 4 against R. solani compared to 3.2, 2.8, 2.6 and 2.8 against F. oxysporum and 2.7, 2.9,3.1 and 3 against R. solani in the wild types T3, T5, S1 and S6,respectively. The growth of T. harzianum isolates (estimated as colony diameter) was significantly increased in all T. harzianum mutants except MT3 and MD5 mutants, CMCase activity was increased from 13.2, 16.2, 10 and 15.8 U/ml / mg protein in the wild isolates T3, T5, S1 and S6 to 18.6, 20.3, 17.7 and 22.5 U/ml / mg protein in the mutants MT3, MT5, MS1 and MS6, respectively. Chitinase activity was also increased in these mutants to 88.4,132.3,86.1 and 136.7 U/ml / mg protein, compared to 82.7, 93.2, 74.5 and 103.2 U/ml / mg protein in their wild, respectively. As comparison between wild types and mutants, the maximum growth was 5.2 and 4.8 cm (as colonies diameters) by MT5 in present of 25% of the recommended field rate of Topsin and Benomyl, compared to 3.3 and 2.2 cm in the wild type T5, respectively, followed by MS6,in which, the maximum growth in this mutant was 4.8 and 4.2 cm in the same concentration of these two fungicides, compared to 3.4 and 2.5 cm in the wild type S6, respectively. According to these results, the mutants MT5 and MS6 developed from UV-irradiation treatment, showed as promising T. harzianum mutants through their antagonistic activity (at mycoparasitism level) , fast growth, higher  CMCase and chitinase activity and growth in 25% of the recommended field rate of Elsa , 25 and 50% of the recommended field rate of Diathen and Mizab and all concentrations of Topsin and Benomyl with complete inhibition of pathogenic fungi F. oxysporum and R. solani.
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