Jin, Na Young
(Department of Applied Biology, College of Agriculture and Life Sciences, Chungnam National University)
,
Lee, You Kyoung
(Department of Biosystems Machinery Engineering, College of Agriculture and Life Sciences, Chungnam National University,)
,
Lee, Bo Ram
(Department of Applied Biology, College of Agriculture and Life Sciences, Chungnam National University)
,
Kim, Yu Seop
(Department of Applied Biology, College of Agriculture and Life Sciences, Chungnam National University)
,
Jun, Jun Hack
(Department of Applied Biology, College of Agriculture and Life Sciences, Chungnam National University)
,
Kim, Hee Ji
(Department of Applied Biology, College of Agriculture and Life Sciences, Chungnam National University)
,
Youn, Young Nam
(Department of Applied Biology, College of Agriculture and Life Sciences, Chungnam National University)
,
Yasunaga-Aoki, Chisa
(Laboratory of Insect Pathology and Microbial Control, Institute of Biological Control, Faculty of Agriculture, Kyushu University)
,
Yu, Youg Man
(Department of Applied Biology, College of Agriculture and Life Sciences, Chungnam National University)
,
Aoki, Chisa
(Laboratory of Insec)
,
Yasunaga, Chisa
A new strain of Bacillus thuringiensis exhibiting activity in Spodoptera exigua was isolated and identified. Synergistic effects appeared when it was mixed with tannic acid, which is a protease inhibitor that enhances insecticidal activities. Substrate reactions with different types of proteases wer...
A new strain of Bacillus thuringiensis exhibiting activity in Spodoptera exigua was isolated and identified. Synergistic effects appeared when it was mixed with tannic acid, which is a protease inhibitor that enhances insecticidal activities. Substrate reactions with different types of proteases were analyzed, and according to the results, trypsin showed proteolytic activity levels of 91.4% and 89.4% on BApNA and BPVApNA substrates, respectively. When trypsin was treated with tannic acid, proteolytic activity levels of 62.2% and 54.5% appeared on BApNA and BPVApNA substrates, respectively, and tannic acid suppressed the trypsin substrate reactions by 29.2% and 34.9%, respectively. Colorimetric activity assays of trypsin identified slightly lower activity levels in larvae that ate a combination of B. thuringiensis KB100 strain and 40 mM of tannic acid, when compared to larvae that died after eating the positive control, control, or the B. thuringiensis subsp. kurstaki KB100. The results of trypsin gene expression level analyses conducted using the qRT PCR analyses indicated that trypsin 1 gene expression levels were lower when 40 mM of tannic acid was added to the B. thuringiensis KB100 strain, compared to the B. thuringiensis KB100 strain alone. Trypsin 2 gene expression levels were also lower when 40 mM of tannic acid was added to the B. thuringiensis KB100 strain. The results of immunofluorescence antibody experiments indicated that the addition of 40 mM of tannic acid to the B. thuringiensis KB100 strain, during decomposition by the mid gut proteases of S. exigua, led to differences in expression levels and image analyses.
A new strain of Bacillus thuringiensis exhibiting activity in Spodoptera exigua was isolated and identified. Synergistic effects appeared when it was mixed with tannic acid, which is a protease inhibitor that enhances insecticidal activities. Substrate reactions with different types of proteases were analyzed, and according to the results, trypsin showed proteolytic activity levels of 91.4% and 89.4% on BApNA and BPVApNA substrates, respectively. When trypsin was treated with tannic acid, proteolytic activity levels of 62.2% and 54.5% appeared on BApNA and BPVApNA substrates, respectively, and tannic acid suppressed the trypsin substrate reactions by 29.2% and 34.9%, respectively. Colorimetric activity assays of trypsin identified slightly lower activity levels in larvae that ate a combination of B. thuringiensis KB100 strain and 40 mM of tannic acid, when compared to larvae that died after eating the positive control, control, or the B. thuringiensis subsp. kurstaki KB100. The results of trypsin gene expression level analyses conducted using the qRT PCR analyses indicated that trypsin 1 gene expression levels were lower when 40 mM of tannic acid was added to the B. thuringiensis KB100 strain, compared to the B. thuringiensis KB100 strain alone. Trypsin 2 gene expression levels were also lower when 40 mM of tannic acid was added to the B. thuringiensis KB100 strain. The results of immunofluorescence antibody experiments indicated that the addition of 40 mM of tannic acid to the B. thuringiensis KB100 strain, during decomposition by the mid gut proteases of S. exigua, led to differences in expression levels and image analyses.
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