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大丽轮枝菌寡糖素诱导的棉花组培细胞的过氧化物酶表达分析

The Peroxidase Isoenzymes of Suspending Culture Cells of Cotton Induced by the Oligosaccharins from Verticillium dahliae

  • 摘要: 将已在察氏液体培养基中培养3d的强毒性(V44)或弱毒性(V64)大丽轮枝菌接种到新鲜的相同培养基中,加入棉花黄萎病感病品种(S)或抗病品种(R)的悬浮培养细胞(或其细胞匀浆).培养4d后收获真菌孢子,用于致萎力测定,收获菌丝体细胞壁用于制备真菌寡糖素(H,来自V44或L,来自V64),然后用该寡糖素诱导悬浮培养的棉花细胞(S或R).棉花细胞的过氧化物酶(POD)同工酶结果表明:(1)所有寡糖素均诱导棉花细胞产生新的过氧化物酶;(2)感病品种豫棉6号的活组培细胞S(或其细胞匀浆)显着影响真菌,用生长过程中接触该活细胞(或其细胞匀浆)的真菌所制备的寡糖素再诱导该活细胞(即S→L(H)→S)时,棉花细胞在36h的POD同工酶谱与其对照,即L(H)→S比较,酶带增减变化显着。致萎力测定结果表明感病品种的活细胞提高了病原毒性和致病力;(3)耐病(豫棉8号)或抗病品种(中棉12号)的活组培细胞R(或其细胞匀浆)对真菌的影响不同于感病品种。R→L(H)→R在36h的POD同工酶谱与L(H)→R比,酶带没有增减变化。但是在H→R或H→S中于9h和24h出现的酶带在R→H→R或R→H→S中分别提前到6h和9h,致萎力测定结果表明抗病品种细胞对病原毒性影响很小。

     

    Abstract: The living cell culture of cotton or its homogenates from V.dahliae-susceptible variety (S) or resistant variety (R) were added into the Czapeks liquid media which had been inoculated 3 d with the higher virulent strain(V44) and lower virulent strain(V64) of V.dahliae respectively.After 4 d additional incubation,the spore of these fungus were collected each to evaluate their wilting or virulence to cotton plants,and the cell wall of the hypha were used to prepare oligosaccharins(H,from V44 and L,from V64)which were as elicitors to induce the cell cultures of cotton.(Sor R) The PAGEspectrums of PODisozymes from cotton cells showed that (1) all oligosaccharins induced cotton cells to produce new PODisozymes; (2)the living cotton cells or its homogenate from susceptible variety affected strongly the fungus.The cotton PODspectrum at 36 h after induction showed that in all the inducing systems,the most outstanding diversity is between S→L(H)→Sand their control,i.e.L(H)→S,i.e.the oligosaccharins from the fungi contacted with the living cotton cells or its homogenate from susceptible variety induced the most strong change of PODisozymes in the living cotton cells from susceptible variety,as compared with the control,L(H)→S.The wilting result showed the living cotton cells from susceptible variety increased the virulence and pathogenicity of the fungi; (3)the impact of living cotton cells or its homogenate from resistant variety on the fungi is different from susceptible one.There is no change of enzyme bands,but the new enzymes occurred at 9 h in H→Rand 24 h in H→Safter induction were ahead to 6 h in R→H→Rand 9 h in R→H→S.The wilting result showed the living cotton cells from resistant variety had little influence on the virulence and pathogenicity of the fungi.

     

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