Isolation, cloning, sequencing and bioinformatic study of a resistance gene analouge against tomato mosaic virus in tow native types of cantaloupe to Iran

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GHARAEI F.; Ghayeb Zamharir M.

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Journal: Journal of Molecular and Cellular Researches Year:2020 | Volume:33 | Issue:1 Page(s): 103-114

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Title

Isolation, cloning, sequencing and bioinformatic study of a resistance gene analouge against tomato mosaic virus in tow native types of cantaloupe to Iran

Author(s)

GHARAEI F. | Ghayeb Zamharir M. | Issue Writer Certificate

Keywords

cantaloupeQ1

resistanceQ2

cloningQ2

PCRQ1

Tomato musaic virusQ2

Abstract

Cucurbita genus belongs to Cucurbitaceae family. These plants are attacked with different pathogenic agents. Currently the effective strategy to control many disease of Cucurbitaceae is the usage of resistant plants. In this study, the NBS domain encoded by resistance genes was studied in Iranian native varieties of cantaloupe. For this purpose, the seeds of Iranian native cantaloupe cultivars were provided and cultivated in greenhouse. DNA was extracted from leaves of different cultivars using CTAB method. Degenerate primers were designed from conserved motives of resistance analogues genes and amplification of these genes was performed using PCR method. The results showed the isolation of an analogous for resistance gene to tomato mosaic virus (ToMV) in cantaloupe cultivars TN-92-99 and TN-92-80 that were cloned in pGEM-T plasmid and then sequenced. Blast analysis indicated that NBS sequence in Irnian cantaloupe share 92% similarity with ToMV resistance gene in watermelon. Phylogenetic tree drawing using resistance gene analogues from cantaloupe and watermelon existing in NCBI gene bank created tow main groups and seven subgroups. Second structure study of protein of ToMV resistance gene analogus using PSIpred software indicated that these proteins only have α helix structure. This protein is secreted in plasma membrane of cantaloupe cultivar TN-92-99 and its extracellular secretion is very little. This is the first study of cloning, expression and identification of enzymatic activity of a resistance gene against ToMV in Iranian native cantaloupe cultivars. Expression of this gene could play important role in resistance against viral diseases including tomato mosaic disease in Cucurbitaceae that could use in breeding programs.

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APA: Copy

GHARAEI, F., & Ghayeb Zamharir, M.. (2020). Isolation, cloning, sequencing and bioinformatic study of a resistance gene analouge against tomato mosaic virus in tow native types of cantaloupe to Iran. JOURNAL OF MOLECULAR AND CELLULAR RESEARCH (IRANIAN JOURNAL OF BIOLOGY), 33(1 ), 103-114. SID. https://sid.ir/paper/375349/en

Vancouver: Copy

GHARAEI F., Ghayeb Zamharir M.. Isolation, cloning, sequencing and bioinformatic study of a resistance gene analouge against tomato mosaic virus in tow native types of cantaloupe to Iran. JOURNAL OF MOLECULAR AND CELLULAR RESEARCH (IRANIAN JOURNAL OF BIOLOGY)[Internet]. 2020;33(1 ):103-114. Available from: https://sid.ir/paper/375349/en

IEEE: Copy

F. GHARAEI, and M. Ghayeb Zamharir, “Isolation, cloning, sequencing and bioinformatic study of a resistance gene analouge against tomato mosaic virus in tow native types of cantaloupe to Iran,” JOURNAL OF MOLECULAR AND CELLULAR RESEARCH (IRANIAN JOURNAL OF BIOLOGY), vol. 33, no. 1 , pp. 103–114, 2020, [Online]. Available: https://sid.ir/paper/375349/en

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