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Specificity of Plant Infection : Molecular Mechanism of Determination of Basic Compatibility by Host-Specific Toxins

植物感染的特异性：宿主特异性毒素确定基本相容性的分子机制

基本信息

批准号：
08456028
负责人：
OTANI Hiroshi
金额：
$ 4.54万
依托单位：
Tottori University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

项目摘要

Certain fungal pathogens are known to produce host-specific toxins (HSTs) as determinants of basic compatibility in plant infection. There are now about twenty pathogens that produce HST.We found that Alternaria brassicicola, cucumber pathotype and tomoto pathotype of Corynespora cassiicola, and Stemphylium vesicarium produce new HSTs named AB-, CCC-, CCT-and SV-toxins, respectively. Unlike other HSTs which are low-molecular-weight secondary metabolites, AB-toxin was a protein and estimated to be 35 kDa.A.brassicicola released AB-toxin from the germinating spores only on host leaves, indicating that AB-toxin production by the pathogen is induced by host-related factors. There are seven pathotypes in A.alternata pathogens. HST (AL-toxin) deficient mutants from tomato to pathotype using REMI method. On the other hand, PCR products were obtained from apple pathotypes, using primers designed from cyclic peptide synthase genes. Detailed molecular analysis of these mutants and PCR products a … More re now in progress. Electrophoreic karyotyping of A.alternata pathotypes were determined by PFGE.In all pathotypes, a chromosomal band with small size of <1.6 Mb was detected commonly. Southern analysis using PCR products from apple pathotype showed that AM-toxin synthesis gene exists in the small chromosome.HSTs (AK-toxin and AM-toxin) produced by Japanese pear and apple pathotypes of A.alternata have primary effects on plasma membranes (PMs) and chloroplasts (Cs) of susceptible cultivars, respectively. Therefore, PMs and Cs were isolated from respective hosts, and interaction between HSTs and proteins in PMs or Cs was analyzed by using a biosensor. Results suggest that AK- and AM-toxin-binding proteins (receptors) exist in PMs and Cs of susceptible cultivars, respectively. Total proteins of apple leaves were analyzed by 2-D gel electrophoresis. A spot of 60 kDa protein with pI of 5.5 was detected only in susceptible cultivars. Isolation and characterization of these proteins is in progress. Less

已知某些真菌病原体会产生宿主特异性毒素（HST），作为植物感染中基本相容性的决定因素。目前大约有二十种病原体能产生HST。我们发现Alternaria brasicicola、黄瓜致病型和Tomoto致病型Corynespora cassiicola以及Stemphylium vesicarium产生新的HST，分别命名为AB-、CCC-、CCT-和SV-毒素。与其他低分子量次生代谢物 HST 不同，AB 毒素是一种蛋白质，估计为 35 kDa。A.brassicicola 仅在寄主叶片上从萌发孢子中释放 AB 毒素，表明病原体产生 AB 毒素是由寄主相关因素诱导的。链格孢病原体有七种致病型。使用 REMI 方法将番茄 HST（AL 毒素）缺陷突变体转化为致病型。另一方面，使用根据环肽合酶基因设计的引物从苹果致病型中获得PCR产物。对这些突变体和 PCR 产物的详细分子分析正在进行中。采用PFGE对链格孢致病型进行电泳核型分析，在所有致病型中，普遍检测到<1.6 Mb的小染色体条带。利用苹果病型PCR产物Southern分析表明，小染色体上存在AM毒素合成基因。日本梨和苹果病型A.alternata产生的HSTs（AK毒素和AM毒素）分别对感病品种的质膜（PMs）和叶绿体（Cs）产生主要影响。因此，从各自的宿主中分离出PM和Cs，并使用生物传感器分析HST与PM或Cs中的蛋白质之间的相互作用。结果表明，AK 和 AM 毒素结合蛋白（受体）分别存在于易感品种的 PM 和 C 中。通过二维凝胶电泳分析苹果叶的总蛋白。仅在敏感品种中检测到 pI 为 5.5 的 60 kDa 蛋白质点。这些蛋白质的分离和表征正在进行中。较少的