TRANSFER OF GLUCANASE GENE TO RESIST LATE BLIGHT DISEASE IN POTATO (Solanum tuberosum L.)

Heba Shebl, E. Metry, M. Rashed, M. A. Abd El-Hafiez, I. Ismail
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Abstract

Potato late blight is a pandemic disease caused by the highly virulent (Phytophthora infestans) fungus. The regeneration capacity was done among three tested potato cultivars (Spunta, Diamont and Desiree). Two different types of explants (inter-node and leaf) were cultured for calli induction and plant regeneration .The highest value of leaf explants was for Desiree cultivar (80%), which induced from developed callus in the 2,4-D medium. The best value of maximum shoot regeneration was also for Desiree cultivar in the BA, IAA and GA3 media from leaf which proved to be more effective. Then cloning of glucanase gene in pRI plasmid which carrying kanamycin resistance (nptII) gene was perfomed and followed by transformation in Agrobacterium tumefaciens LBA4404 strain which used for plant transfection. Nucleotide and amino acid sequences of transformed Agrobacterium were analyzed. The putative transgenic plantlets genomes and cDNA of the isolated RNA for glucanase gene.
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马铃薯抗晚疫病的谷氨酰胺酶基因转移
马铃薯晚疫病是一种由高毒力真菌(疫霉)引起的大流行疫病。对3个马铃薯品种(Spunta、Diamont和Desiree)的再生能力进行了研究。用两种不同类型的外植体(节间和叶片)诱导愈伤组织和植株再生,其中叶片外植体在2,4- d培养基上的诱导率最高(80%)。在叶片BA、IAA和GA3培养基上,Desiree品种的最大芽再生值也最高,且效果更好。然后在携带卡那霉素抗性(nptII)基因的pRI质粒上克隆葡聚糖酶基因,并将其转化为农杆菌LBA4404菌株进行植物转染。对转化农杆菌的核苷酸和氨基酸序列进行了分析。推定的转基因植株基因组和分离的葡聚糖酶基因RNA cDNA。
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