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程國軍*,羅莎,謝婧,吳和濤.豌豆根瘤菌RND外排轉運基因tpaA的功能研究[J].中南民族大學學報自然科學版,2019,(2):199-203
豌豆根瘤菌RND外排轉運基因tpaA的功能研究
Function of efflux RND transporter gene tpaA in Rhizobium leguminosarum
  
DOI:10.12130/znmdzk.20190209
中文關鍵詞: 豌豆根瘤菌  外排轉運基因  生長  抗生素耐藥性  共生固氮
英文關鍵詞: Rhizobium leguminosarum  efflux RND transporter gene  growth  antibiotic resistance  symbiotic fixation
基金項目:國家自然科學基金資助項目(31772399);中央高?;究蒲袠I務費專項資金資助項目(CZY18022)
作者單位
程國軍*,羅莎,謝婧,吳和濤 中南民族大學 生命科學學院/微生物資源與利用湖北省工程技術研究中心,武漢 430074 
摘要點擊次數: 192
全文下載次數: 189
中文摘要:
      Rhizobium. leguminosarum 3841 tpaA基因編碼RND家族內膜外排轉運蛋白,RND外排泵的過量表達是革蘭陰性菌細菌抗生素耐藥性形成的重要原因之一. 采用基因同源重組構建豌豆根瘤菌tpaA基因突變株RLtpaA,并研究了該基因突變對根瘤菌生長、抗生素耐藥性以及共生固氮的影響. 結果表明,tpaA基因突變會使根瘤菌在AMS基本培養基生長延遲,但不影響其在TY天然培養基中的正常生長. tpaA基因突變可能抑制了根瘤菌中的RND家族外排泵的過量表達,從而使其抗生素耐藥性降低. 接種了tpaA基因突變株的豌豆植株能形成紅色有效根瘤,但其接種根瘤固氮酶活性降低. 實時熒光定量RT-PCR分析表明tpaA基因在根瘤類菌體中的表達顯著上調.
英文摘要:
      In Rhizobium leguminosarum 3841, tpaA gene encodes RND intima efflux transporter protein. Overexpression of RND family efflux pumps is one of the important reasons for the formation of antibiotic resistance in Gram-negative bacteria. R. leguminosarum tpaA mutant RLtpaA was constructed through homologous recombination, and the effects of tpaA gene mutation on growth, antibiotic resistance and symbiotic fixation were further studied. The results showed that the mutation of tpaA gene delayed the bacterial growth in minimal medium AMS, but did not affect the normal growth in natural medium TY. Antibiotic resistance experiments showed that tpaA gene mutation may inhibit the overexpression of RND family efflux pumps in R. leguminosarum, thus reducing its antibiotic resistance. Symbiotic experiments showed that pea plants inoculated with tpaA gene mutant formed effective red nodules, but can lead to a?decline of the nitrogen fixation ability. Quantitative real-time RT-PCR result showed that tpaA gene expression can be induced under symbiotic condition.
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