CAJ | 학술논문

为了明确含有cry8E基因的绿僵菌Metarhizium anisopliae不同转化子存在毒力差异的原因，揭示胞外酶与毒力之间的关系，本研究测定了6株含cry8E基因绿僵菌阳性转化子的酸性蛋白酶、中性蛋白酶、碱性蛋白酶、几丁质酶和磷脂酶活性及其对暗黑鳃金龟幼虫Holotrichia parallela毒力，并分析了各个酶活指标与菌株毒力之间的相关性。生测结果表明，不同转化子间毒力差异较大，LT50的变化范围为14.48～21.26 d，校正累计死亡率变化范围为64.50%～92.86%。酶活测定结果表明，不同菌株之间活力差异较大，且不同酶的活力变化规律不一致。将不同转化子的酶活分别与校正累积死亡率和 LT50值进行一元回归分析，发现各转化菌株酸性蛋白酶和几丁质酶与毒力相关显著。为了进一步确定酸性蛋白酶和几丁质酶对毒力的综合影响，将不同转化子的酶活与校正累积死亡率、LT50进行二元回归分析，发现仅有几丁质酶活性与菌株毒力相关性显著。因此，推测不同转化子对寄主毒力存在差异主要是由于菌株间几丁质酶活性不同，几丁质酶可以作为大量转化子初步筛选的参考性指标，但是不能完全替代。
위료명학함유cry8E기인적록강균Metarhizium anisopliae불동전화자존재독력차이적원인，게시포외매여독력지간적관계，본연구측정료6주함cry8E기인록강균양성전화자적산성단백매、중성단백매、감성단백매、궤정질매화린지매활성급기대암흑새금구유충Holotrichia parallela독력，병분석료각개매활지표여균주독력지간적상관성。생측결과표명，불동전화자간독력차이교대，LT50적변화범위위14.48～21.26 d，교정루계사망솔변화범위위64.50%～92.86%。매활측정결과표명，불동균주지간활력차이교대，차불동매적활력변화규률불일치。장불동전화자적매활분별여교정루적사망솔화 LT50치진행일원회귀분석，발현각전화균주산성단백매화궤정질매여독력상관현저。위료진일보학정산성단백매화궤정질매대독력적종합영향，장불동전화자적매활여교정루적사망솔、LT50진행이원회귀분석，발현부유궤정질매활성여균주독력상관성현저。인차，추측불동전화자대기주독력존재차이주요시유우균주간궤정질매활성불동，궤정질매가이작위대량전화자초보사선적삼고성지표，단시불능완전체대。
In this study, we tested the acid protease, neutral protease, alkaline protease, chitinase, phospholipase enzyme activities of six Metarhizium anisopliae transformants with cry8E gene insertion and virulence against Holotrichia parallela in order to clarify the correlationship between extracellular enzyme activity and virulence of each strain. The results showed that the virulence among six transformants assayed was significantly different, and the ranges of the estimated LT50 and the corrected cumulative mortality were 14.48~21.26 d, 64.50%~92.86%, respectively. There also has significant difference in the enzyme activities among six transformants. Regression analysis also revealed that acid protease and chitinase were significantly correlated to the corrected cumulative mortality and LT50. Binary regression analysis of combination effects of acid protease and chitinase on virlence also revealed that only chitinase was significantly correlated to the corrected cumulative mortality and LT50. It suggested that the difference of virulence among transformants may be mainly derived from the activity of chitinase, and it could be used as virulence index for large scale of early candidate transformants selection, however, but it could not completely replace the virulence measurement in future.