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| Optimization of preparation method of Bacillussubtilis strain R31 protoplast |
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| DOI： |
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| Abstract: |
| Bacillus subtilis…strain…R31…could…be…used…to…control…banana…Fusarium…Wilt…in…fields， and…could…not…be…
genetic…operated…by…conventional…preparation…of…competent…cells， so…researching…its…genetic…transformation…method…could…
be…benefit…for…the…understanding…of…its…molecular…bio-control…mechanism.…Based…on…the…growth…curve…and…growing…status…of…
strain…R31…in…LB…media， the…culture…time…suit…for…protoplast…preparation…and…regeneration…was…chosen.…The…growth…curve…
study…showed…that…in…LB…media， the…growth…of…R31…was…in…the…logarithmic…phase…from…3…h…to…6…h…after…inoculated， and…it…
was…at…the…end…of…logarithmic…phase…after…4.5…h.…After…7…h， the…growth…of…R31…was…in…the…stationary…phase.…Observations…by…
microscopy…discovered…that…R31…growing…as…free-floating…planktonic…mode…at…the…early…stage…of…the…logarithmic…growth，
but…bacteria…began…to…connect…with…each…other…after…5.5…h…and…gradually…formed…a…net…structure， then…formed…a…three-
dimensional…structure…at…8…h.…Free-floating…planktonic…cells…were…suit…for…the…protoplast…preparation…and…regeneration， but…
gave…consideration…to…the…concentration…of…cells…for…protoplast…preparation， the…optimized…culture…time…was…4-4.5…h， under…
this…condition， and…the…best…lysozyme…concentration…was…3.5…mg/mL. |
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