發(fā)布時間：2018-10-25 14:16

【摘要】：為了明確球孢白僵菌Beauveria bassiana 3菌株TST05、FDB01、SYN01對栗實(shí)象甲老熟幼蟲的感染、致死率、致死中時、以及胞外蛋白酶、幾丁質(zhì)酶、脂肪酶的活性與毒力的關(guān)系,本研究采用3個菌株的孢子懸浮液(1.0×108孢子/m L),浸蟲法感染栗實(shí)象甲幼蟲,統(tǒng)計連續(xù)8 d的校正死亡率;以栗實(shí)象甲幼蟲的蟲體作為菌株的唯一碳源制備液體培養(yǎng)基,測定培養(yǎng)過程中各菌株的脂肪酶、幾丁質(zhì)酶、類枯草桿菌蛋白酶(Pr1酶)連續(xù)8 d的酶活性變化。結(jié)果表明,球孢白僵菌3個菌株TST05、FDB01、SYN01感染栗實(shí)象甲幼蟲后,1-4 d內(nèi)表現(xiàn)出染病和死亡的癥狀,連續(xù)8 d累積校正死亡率分別為40%、44.4%、62.22%,菌株SYN01對栗實(shí)象甲的致死率最高,與其他兩菌株的差異顯著;菌株TST05、FDB01、SYN01的Pr1酶活性最大峰值分別為(175.56±0.7)U/m L、(172.74±2.32)U/m L、(195.71±5.41)U/m L。幾丁質(zhì)酶活性最大峰值分別為(12.58±0.58)U/m L、(13.06±1.16)U/m L、(15.12±0.32)U/m L。都是菌株SYN01的酶活性最大,而TST05和FDB01之間差異不顯著;球孢白僵菌3個菌株的Pr1酶、幾丁質(zhì)酶、脂肪酶在8 d內(nèi)的酶活性平均值與染菌幼蟲8 d累積校正死亡率的線性回歸方程分別為y=0.4641x+122.45(R2=0.8854)、y=0.034x+10.473(R2=0.9328)、y=0.0354x+2.4586(R2=0.1201),說明菌株的Pr1酶和幾丁質(zhì)酶活性與幼蟲死亡率之間呈顯著的線性相關(guān),而菌株的脂肪酶活性與菌株對幼蟲的致死率不具有線性關(guān)系。綜合分析,菌株SYN01可以作為生物防治栗實(shí)象甲的病原菌種;Pr1酶與幾丁質(zhì)酶的活性可作為菌株的毒力因子。
[Abstract]:In order to clarify the relationship between the infection, mortality, death rate, the activity of extracellular protease, chitinase, lipase and virulence of Beauveria bassiana 3 strain of Beauveria bassiana on mature larvae of chestnut. In this study, the sporozoite suspension (1.0 脳 108spore / m L), dipping method) of three strains was used to infect chestnut larvae, and the corrected mortality of chestnut larvae was calculated for 8 days, and the liquid medium was prepared by using the larvae of chestnut larvae as the sole carbon source of the strains. Lipase, chitinase and Pr1 enzyme activity of each strain were determined for 8 days. The results showed that three strains of Beauveria bassiana (TST05,FDB01,SYN01) showed symptoms of infection and death within 1-4 days after infection with C. bassiana larvae. The cumulative corrected mortality for 8 days was 400.44.4% and 62.22%, respectively. Strain SYN01 had the highest mortality to chestnut weevil. The peak value of Pr1 enzyme activity in TST05,FDB01,SYN01 was (175.56 鹵0.70) U / -mL, (172.74 鹵2.32) U / -mL, (195.71 鹵5.41) UP / mL, respectively. The peak value of chitinase activity was (12.58 鹵0.58) U / mL, (13.06 鹵1.16) U / mL, (15.12 鹵0.32) U / mL, respectively. The enzyme activity of SYN01 was the largest, but there was no significant difference between TST05 and FDB01. The Pr1 enzyme, chitinase of three strains of Beauveria bassiana, The linear regression equations of lipase activity in 8 days and the cumulative corrected mortality rate in 8 days of inoculated larvae were yx0.4641x 122.45 (R2O0.8854), YC0.034x 10.473 (R20.9328), YC0.0354x 2.4586 (R2C0.1201), indicating that there was a significant linear correlation between the activity of Pr1 enzyme and chitinase activity and larval mortality. However, there was no linear relationship between lipase activity and larval mortality. Comprehensive analysis showed that strain SYN01 could be used as pathogen of chestnut weevil, and the activity of Pr1 and chitinase could be used as virulence factor of chestnut.
【作者單位】： 山西大學(xué)生命科學(xué)學(xué)院;
【基金】：山西省科技攻關(guān)項(xiàng)目(20140311017-6)
【分類號】：S436.64;S476.12

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