EXPRESSION OF ANIMAL INTERFERON(FN) SYSTEM-LIKE METABOLISM IN PLANTS AND MOLECULAR ANALYSIS OF ANTI-VIRAL MECHANISM IN TRANSGENIC PLANTS INTRODUCED INF-RELATING GENES.

动物干扰素(FN)系统类代谢在植物中的表达及转基因植物抗病毒机制的分子分析引入INF相关基因。

• 批准号: 06454136
• 负责人: EHARA Yoshio
• 金额: $ 4.74万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06454136/
• 关键词: plant virus; resistance; interferon; 2,5-A synthetase; protein kinase; transgenic plants; 植物ウイルス抵抗性; 動物インターフェロン

This experiment was carried out to confirm whether or not the mammalian inteferon (IFN) system-like metabolism are induced in virus infected plants. Although the extracts from necrotic local lesion-formed leaves on cucumber mosaico virus (CMV)-infected cowpea and tobacc mosaic virus (TMV)-infected N-gene-carrying tobacco were analyzed by Western blotting with antisera of mammalian alpha, beta and gamma interferons, any positive signals were not detected. To examine the occurrence of the 2,5-oligoadenylate synthetase (2,5-AS) and protein kinase (PX) induced in animal cells by IFN in these infected plants, the extracts from the plants were analyzed by Northern hybridization using mammalian 2,5-AS and PK cDNAs as probes. 2,5-AS and PK mRNAa were not detected in the extracts. The 2,5 AS activity was also examined using the sensitive radioimmune assay, but no significant activity was observed. From the rusults, it was concluded that the mammalian interferon-like metabolic system was absent in plants. A human 2,5-AS cDNA was introduced into tobacco (Nicotiana tabacum cv. Samsun NN) plants by Agrobacterium-mediated transformation. The transgenic plants expressed the 2,5-AS mRNA,but not the 2,5-AS protein. While the transgenic plants showed resistance to CMV and TMV.The resistance is probably induced by co-suppression between 2,5-AS mRNA and virus RNA.On the other hand, the transgenic tobacco plants introduced IFN-induced PK cDNA also showed resistance to both viruses. In this case, neither PK-mRNA nor PK were detected in the transgenic plants, suggesting the resistance caused by indirect mechanism for a foreign cDNA integration in to the host DNA.As mentioned above, it was confirmed that INF-like metabolism was not present in plants, and that virus resistance in transgenic plants introduced IFN-related enzyme genes was resulted in different mechanism from IFN system.

进行该实验是为了确认在病毒感染的植物中是否诱导哺乳动物干扰素（IFN）系统样代谢。尽管用哺乳动物α、β和γ干扰素的抗血清对感染黄瓜花叶病毒(CMV)的豇豆和感染烟草花叶病毒(TMV)的携带N基因的烟草的局部坏死病斑叶子提取物进行蛋白质印迹分析，但没有检测到任何阳性信号。为了检查这些受感染植物中 IFN 在动物细胞中诱导的 2,5-寡腺苷酸合成酶 (2,5-AS) 和蛋白激酶 (PX) 的发生情况，使用哺乳动物 2,5-AS 和 PK cDNA 作为探针，通过 Northern 杂交分析植物提取物。提取物中未检测到 2,5-AS 和 PK mRNAa。还使用灵敏的放射免疫测定法检查了 2,5 AS 活性，但没有观察到显着的活性。从结果可以得出结论，植物中不存在哺乳动物干扰素样代谢系统。通过农杆菌介导的转化将人类 2,5-AS cDNA 引入烟草（Nicotiana tabacum cv. Samsun NN）植物中。转基因植物表达2,5-AS mRNA，但不表达2,5-AS蛋白。而转基因植物表现出对CMV和TMV的抗性。这种抗性可能是由2,5-AS mRNA和病毒RNA之间的共抑制诱导的。另一方面，引入IFN诱导的PK cDNA的转基因烟草植物也表现出对两种病毒的抗性。在这种情况下，在转基因植物中既没有检测到PK-mRNA也没有检测到PK，这表明抗性是由外源cDNA整合到宿主DNA中的间接机制引起的。如上所述，证实植物中不存在类INF代谢，并且引入IFN相关酶基因的转基因植物中的病毒抗性是由与IFN系统不同的机制导致的。

项目成果

NAKAMURA,S., YAMAUCHI.N., HASE,S., KARASAWA,A., HASHIBA,T., AND EHARA,Y.: "VIRUS RESISTANCE IN TRANSGENIC TOBACCO PLANTS INTRODUCED INTERFERON-INDUCED PROTEIN KINASE GENE." TOHOKU JOUR.AGRIC.RES.48(in preparation). (1997)

NAKAMURA,S.、YAMAUCHI.N.、HASE,S.、KARASAWA,A.、HASHIBA,T. 和 EHARA,Y.：“转基因烟草植物中的病毒抗性引入了干扰素诱导的蛋白激酶基因。”

Nakamura,S.et al.: "Plants transformed with mammalian2,5 oligoadanylate synthetase gene show resistance to virus infection" Ann.Phytopath.Soc.Japan. 60. 691-693 (1994)

Nakamura,S.et al.：“用哺乳动物 2,5 寡腺苷酸合成酶基因转化的植物显示出对病毒感染的抵抗力”Ann.Phytopath.Soc.Japan。

Nakamura,S.et al.: "Plants transformed with mammalian 2',5' oligoadanylate synthetase gene show resistance to virus infection" Ann. Phytopath. Soc. Japan. 60. 691-693 (1994)

Nakamura,S.et al.：“用哺乳动物 2,5 寡腺苷酸合成酶基因转化的植物显示出对病毒感染的抵抗力”Ann。

NAKAMURA,S., YOSHIKAWA,M., TAIRA,H., AND EHARA,Y.: "PLANTS TRANSFORMED WITH MAMMALIAN 2,5 OLIGOADENYLATE SYNTHETASE GENE SHOW RESISTANCE TO VIRUS INFECTION." ANN.PHYTOPATH.SOC.JAPAN. 60. 691-693 (1994)

NAKAMURA,S.、YOSHIKAWA,M.、TAIRA,H. 和 EHARA,Y.：“用哺乳动物 2,5 寡腺苷酸合成酶基因转化的植物显示出对病毒感染的抵抗力。”

EHARA,Y., TAIRA,H., NAKAMURA,S., YAMAUCHI,N., SHIRASAWA,N., KARASAWA,A., AND HASE,S.: "ANIMAL INTERFERON SYSTEM-LIKE METABOLISM IS ABSENT IN PLANTS." ANN.PHYTOPATH.SOC.JAPAN. 63(in preparation). (1997)

EHARA,Y.、TAIRA,H.、NAKAMURA,S.、YAMAUCHI,N.、SHIRASAWA,N.、KARASAWA,A. 和 HASE,S.：“植物中不存在动物干扰素系统样代谢。”