U. S. -Japan joint research on the molecular pathology of host-specific-toxin-depending fungal infection to plants

美日联合研究植物宿主特异性毒素依赖性真菌感染的分子病理学

• 批准号: 02044099
• 负责人: KOHMOTO Keisuke
• 金额: $ 9.41万
• 依托单位: Tottori University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for international Scientific Research
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1991
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02044099/
• 关键词: Host-specific toxin; Host-selective toxin; Infection-inducing factor; Toxin receptor; Detoxifying enzyme; Toxin-synthesizing enzyme; Transformation system; Gene for toxin production; 宿生特異的毒素; 毒素生成酵素; 毒素解毒酵素; 病原性遺伝子; 形質転換系; Alternaria; Helmin

l. Occurrence of AM-toxin an ACT-toxin producing Alternaria Alternata in North America : The pathogen for new alternariosis on Delicious lines in North Carolina apple orchards was found to produce AM-toxin I by bioassay and HPLC analysis, and to fall into the apple pathotype of Alternaria alternate.2. Biochemical analysis revealed that although AK-, ACT-, AM-, ACR- and HMT-toxins have different primary targets in susceptibility genotypes, they exert in common an ability to induce host-specifically the accessibility to invader by nullifying dynamic defense mechanisms in plant cells.3. Pyricularia oryzae causing rice blast produced an infection-inducing factor on germination, which may involve in establishment of the basic compatability between rice plants and the fungus.4. Detection and isolation of toxin receptors : (1) Covalent binding sites of victorin (HV-toxin) in oat leaf tissues were detected by polyclonal, antibodies : in vivo binding of victorin to proteins of 100 and 45 Kd was … More observed in both susceptible and resistanit cultivars of oats, and also, in vitro binding to 100, 55 and 45 Kd in both genotypes. (2) AK-toxin was found to bind to plasma membranes purified from susceptible cultivar, but not those from resistant, of Japanese pear. To confirm, polyclonal anti-AK-toxin antibodies are raising in rabbits.5. Cochliobolus carbonum race 1 possesses two multifunctional enzymes required for HC-toxin cyclic peptide synthesis, HTS-1 & HTS-2, which activate-certain member amino acids by ATP/PPI exchange. A partial CDNA clone, encoding HTS-1 (220 kd) was identified using a specific polyclonal anti-HTS-1 antibody and by hybridization, to a synthetic oligonucleotide corresponding to the amino acid sequence of a tryptic peptide of the enzyme. The gene for HTS-1 is part of a contiguous 22 kb of DNA that is present in isolates of the fungus that make HC-toxin and is lacking in those that do not.6. An efficient integrative transformation system (PDH25rla) for Alternaria alternata was established by using a plasmid vector PDH25 and a highly repetitive ribosomalRNA gene cluster (rDNA) of A. alternata. The transformation occurred by homologous recombination as a single crossover between the plasmid-borne RDNA segment and its homologue in the chromosome.7. HC-toxin reductase that inactivates HC-toxin at the essential carbonyl group, is detectable only in extracts of maize that are resistant to C. carbonium race 1 (genotype Hm/Hm or Hm/hm). Less

L.一种产生ACT毒素的链格孢菌AM毒素在北美的发生情况：通过生物测定和HPLC分析，发现北卡罗来纳州苹果园Delicious品系上的新链格孢病病原菌产生AM毒素I，属于苹果链格孢菌致病型.生化分析表明，尽管AK-、ACT-、AM-、ACR-和HMT-毒素在感病基因型中的主要作用靶点不同，但它们都具有通过破坏植物细胞的动态防御机制来诱导寄主特异性的入侵者进入的能力.稻瘟病菌在萌发过程中产生一种侵染诱导因子，这可能与稻瘟病菌与水稻之间基本亲和性的建立有关.毒素受体的检测与分离：（1）通过多克隆抗体检测燕麦叶组织中Victorin（HV-毒素）的共价结合位点：Victorin与100和45 Kd蛋白的体内结合， ...更多信息 在感病和抗病燕麦品种中均观察到了与100、55和45 Kd的结合。(2)AK-毒素被发现绑定到质膜纯化的感病品种，而不是那些从抗性，日本梨。为了证实，多克隆抗AK毒素抗体在兔子中产生。Cochliobolus carbonum race 1具有两个多功能酶HTS-1和HTS-2，这两个酶是合成HC-毒素环肽所必需的，它们通过ATP/PPI交换来激活某些成员氨基酸。使用特异性多克隆抗HTS-1抗体并通过与对应于酶的胰蛋白酶肽的氨基酸序列的合成寡核苷酸杂交来鉴定编码HTS-1（220 kd）的部分cDNA克隆。HTS-1基因是一个连续的22 kb DNA的一部分，该DNA存在于产生HC毒素的真菌分离株中，而不产生HC毒素的真菌分离株中则缺乏该基因。以烟草赤星病菌（Alternaria alternata）为研究对象，利用质粒载体PDH 25和高重复核糖体RNA基因簇（rDNA）构建了烟草赤星病菌（Alternaria alternata）的高效整合转化体系PDH 25 rla。交替转化是通过质粒携带的rDNA片段与其在染色体中的同源物之间的单交换进行的同源重组。HC-毒素还原酶使HC-毒素在必需羰基上失活，仅在抗C.碳离子小种1（基因型Hm/Hm或Hm/hm）。少

项目成果

小林 裕和: "ハイブリダイゼ-ション:原理と応用" 植物細胞工学. 2. 543-555 (1990)

Hirokazu Kobayashi：“杂交：原理和应用”植物细胞工程 2. 543-555 (1990)。

Kohmoto,K.: "Host recognition by toxigenic plant pathogens" Experientia. 47. 755-764 (1991)

Kohmoto,K.：“产毒植物病原体的宿主识别”Experientia。

Meeley,R.B.: "A biochemical phenotype for a disease resistnace gene of maize" Plant Cell.

Meeley,R.B.：“玉米抗病基因的生化表型”植物细胞。

Tsuge, T. et al.: "Efficient integrative transformation of the phytopathogenic fungus Alternaria alternata mediated by the repetitive rDNA sequences." Gene. 90. 207-214 (1990)

Tsuge, T. 等人：“通过重复的 rDNA 序列介导植物病原真菌 Alternaria alternata 的高效整合转化。”

Ngernprasirtsiri, J. et al.: "Application of an efficient strategy with a phage lambda vector for constructing a physical map of the amyloplast genome of sycamore (Acer pseudoplatanus)." Archives of Biochemistry and Biophysics. 276. 172-179 (1990)

Ngernprasirtsiri, J. 等人：“应用噬菌体 lambda 载体的有效策略来构建美国梧桐（Acer pseudoplatanus）淀粉体基因组的物理图谱。”