CAJ | 학술논문

研究以苄嘧磺隆降解菌Rhodococcus sp. BX2与丁草胺降解菌Acinetobacter sp. LYC-1为亲本，优化用于构建可同时降解苄嘧磺隆与丁草胺的融合子的原生质体融合条件。通过单因素试验和正交试验，确定原生质体融合的最佳条件：PEG400040%、35℃、10 min，500μL新生磷酸钙溶液，pH 7.5，此条件下融合频率达到2.67×10-7。以青霉素和磷霉素作为筛选的遗传标记，最终获得在含有上述两种除草剂的无机盐培养基中可稳定继代培养8代以上的融合子F1。该融合子在含有100 mg·mL-1苄嘧磺隆和100 mg·mL-1丁草胺的无机盐培养基中，F1的降解率分别为65.35％和62.41％。
연구이변밀광륭강해균Rhodococcus sp. BX2여정초알강해균Acinetobacter sp. LYC-1위친본，우화용우구건가동시강해변밀광륭여정초알적융합자적원생질체융합조건。통과단인소시험화정교시험，학정원생질체융합적최가조건：PEG400040%、35℃、10 min，500μL신생린산개용액，pH 7.5，차조건하융합빈솔체도2.67×10-7。이청매소화린매소작위사선적유전표기，최종획득재함유상술량충제초제적무궤염배양기중가은정계대배양8대이상적융합자F1。해융합자재함유100 mg·mL-1변밀광륭화100 mg·mL-1정초알적무궤염배양기중，F1적강해솔분별위65.35％화62.41％。
In this study, functional fusants with dual functions to simultaneously degrade bensulfuron-methyl and butachlor were constructed through protoplast fusion of Rhodococcus sp. BX2 and Acinetobacter sp.LYC-1. The protoplast fusion condition was optimized and finally the fusion frequency reached to 2.67 × 10-7 under the following condition:40%of PEG4000, 10 min, 35℃, 500μL of freshly-prepared calcium phosphate, pH 7.5. The fusant F1 was obtained with the use of penicillin G and fosfomycin as selection markers and determined to be able to pass more than eight generations stablely on the plates containing bensulfuron-methyl and butachlor. The fusant F1 had respective high degradation rate of 65.35%BSM and 62.41%butachlor in mineral medium supplemented by BSM 100 mg·L-1 and butachlor 100 mg·L-1.