對已衰退的菌種(群體)進(jin)行(xing)純(chun)種(zhong)分(fen)離(li)和(he)選(xuan)擇(ze)性(xing)培(pei)養(yang),使(shi)其(qi)中(zhong)未(wei)衰(shuai)退(tui)的(de)個(ge)體(ti)獲(huo)得(de)大(da)量(liang)繁(fan)殖(zhi),重(zhong)新(xin)成(cheng)為(wei)純(chun)種(zhong)群(qun)體(ti)的(de)措(cuo)施(shi)。狹(xia)義(yi)的(de)複(fu)壯(zhuang)是(shi)一(yi)消(xiao)極(ji)措(cuo)施(shi),一(yi)般(ban)指(zhi)對(dui)已(yi)衰(shuai)退(tui)的(de)菌(jun)種(zhong)進(jin)行(xing)複(fu)壯(zhuang);guangyidefuzhuangshiyijijidecuoshi,jizaijunzhongdeshengchanxingzhuangweishuaituiqianjiubuduanjinxingchunzhongfenliheshengchanxingzhuangceding,yizaiquntizhonghuodeshengchanxingzhuanggenghaodezifatubianzhu。
1、定義
菌(jun)種(zhong)退(tui)化(hua)是(shi)指(zhi)群(qun)體(ti)中(zhong)退(tui)化(hua)細(xi)胞(bao)在(zai)數(shu)量(liang)上(shang)占(zhan)一(yi)定(ding)比(bi)例(li)後(hou),所(suo)表(biao)現(xian)出(chu)群(qun)體(ti)性(xing)能(neng)變(bian)差(cha)的(de)現(xian)象(xiang)。因(yin)此(ci),在(zai)已(yi)經(jing)退(tui)化(hua)的(de)群(qun)體(ti)中(zhong),仍(reng)然(ran)有(you)一(yi)定(ding)數(shu)量(liang)尚(shang)未(wei)退(tui)化(hua)的(de)個(ge)體(ti)。
2、複壯的方法
純種分離法
zaishuaituijunzhongdexibaoqunzhong,yibanhaicunzaizherengbaochiyuanyoudianxingxingzhuangdegeti。tongguochunzhongfenlifa,shefabazhezhongxibaotiaoxuanchulaijikedadaofuzhuangdexiaoguo。chunzhongfenlifangfajiduo,datikefenweilianglei,yileijiaocufang,kedadao“菌落純”水平(pure culture in colony level);另一類較精細,可達到“菌株純”的水平(pure culture in strain level),現表解如下:
| 菌落純 | 平板表麵塗布法 |
| 平板劃線分離法 | |
| 瓊脂培養基澆注法 | |
| 細胞純 | 用“分離小室”進行單細胞分離 |
| 用顯微操縱器進行單細胞分離 | |
| 用菌絲尖端切割法進行單細胞分離 | |
| 用激光鑷子技術從毛細管中分離 |
通過宿主體內複壯
對於因長期在人工培養基上移種傳代而衰退的病原菌,可接種到相應的昆蟲或動、植物宿體中,通過這種特殊的“選擇性培養基”一至多次選擇,就可從典型的病灶部位分離到恢複原始毒力的複壯菌株。例如,經長期人工培養的Bacillus thuringiensis會發生毒力減退和殺蟲效率降低等衰退現象。這時,就可將已衰退的菌株去感染菜青蟲的幼蟲,然後再從最早、最嚴重罹病的蟲體內重新分離出產毒菌株。
淘汰已衰退的個體
有人發現,若對S. microflavus “5406”農用抗生菌的分生孢子采用-10℃~-30℃的低溫處理5~7天,使其死亡率達到80%左右。結果會在抗低溫的存活個體中留下未退化的個體,從而達到了複壯的效果。
3、shiyongqingkuangbianjiyibanqingkuangxia,duiyushengchanqiye,junzhongguanlidouyaoqiuduijunzhongjinxingdingqidefenlichunhua,yejiushishuofenlishaixuanyibandoushizaituihuazhiqianjiujinxingdeyizhongchangguigongzuo,shifangzhijunzhongtuihuahuozheshengchanyuzhongercaiqudeyizhongjijicuoshi。lingwai,zaichuxianfajiaoshuipingjiaogaohejiaodideqingkuangxia,yehuijinxingjunzhongdefenlishaixuan。qianzhedemudeshiqiwangcongfajiaoyezhongfenlichugaochanjunzhu,erhouzhezeshiqiwangcongkenengyijingtuihuadezhongzizhongfenlichushangweituihuadejunzhu。
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