CAJ | 학술논문

通过农杆菌介导法将抗真菌的几丁质酶基因转入洋桔梗，以提高其对真菌病害的抗性。结果表明，成功克隆了抗真菌活性较强的菜豆几丁质酶基因，构建了植物表达载体，并对洋桔梗Double Mariachi Pink叶块进行转化，获得了卡那霉素抗性植株，对抗性植株进行了2次PCR检测( nptⅡ基因)，第1次PCR检测获得了13个阳性株系，培养60 d后再进行第2次PCR检测，获得11个阳性株系，再对生长良好的5个转基因株系进行RT-PCR检测，获得了3个阳性株系。3个阳性株系的几丁质酶平均活性(0.215 U、0.225 U和0.286 U)均显著高于未转化植株(0.133 U)，转基因株系几丁质酶粗提液对大豆链霉菌5A、5E和灰葡萄孢菌3种指示真菌的抑菌圈直径均显著大于未转化植株，其中抑菌效果最好的株系对上述3种指示真菌的抑菌圈直径与对照相比，分别增大了106.00％、90.91％和71.53％。
통과농간균개도법장항진균적궤정질매기인전입양길경，이제고기대진균병해적항성。결과표명，성공극륭료항진균활성교강적채두궤정질매기인，구건료식물표체재체，병대양길경Double Mariachi Pink협괴진행전화，획득료잡나매소항성식주，대항성식주진행료2차PCR검측( nptⅡ기인)，제1차PCR검측획득료13개양성주계，배양60 d후재진행제2차PCR검측，획득11개양성주계，재대생장량호적5개전기인주계진행RT-PCR검측，획득료3개양성주계。3개양성주계적궤정질매평균활성(0.215 U、0.225 U화0.286 U)균현저고우미전화식주(0.133 U)，전기인주계궤정질매조제액대대두련매균5A、5E화회포도포균3충지시진균적억균권직경균현저대우미전화식주，기중억균효과최호적주계대상술3충지시진균적억균권직경여대조상비，분별증대료106.00％、90.91％화71.53％。
An antifungal chitinase gene was transferred into the leaf segments of lisianthus [ Eustoma grandiflorum ( Raf. ) Shinn. ] Double Mariachi Pink to enhance the resistance to fungal diseases by the Agrobacterium-mediated method. Thirteen kanamycin-resistant lines were obtained and analyzed by PCR, among which, three transgenic lines were con-firmed nptII gene positive by RT-PCR. The chitinase activities of the three transgenic lines were 0. 215 U, 0. 225 U and 0. 286 U, significantly higher than the non-transformed line (0. 133 U). The inhibitory effects (in terms of the diameter of inhibition zone) on three fungi ( Streptomyce 5A, 5E and boytrytis cinerea) were significantly greater than the non-trans-formed line;The strongest inhibitory transgenic line showed larger diameters of inhibition zone by 106. 00%, 90. 91% and 71. 53%.