Articles
DIFFERENTIAL DISPLAY SCREENING OF GENES INVOLVED IN DROUGHT AND SALT TOLERANCE IN THREE CULTIVARS OF PRUNUS MUME SIEB. ET ZUCC.
Article number
763_34
Pages
259 – 266
Language
English
Abstract
Prunus mume cv. ‘Nanko’, which is the leading cultivar in Japan for making pickled ume, is a special product in Wakayama prefecture.
Recent warm, dry weather and salt-contaminated irrigation have resulted in poor field establishment of stocks and decreased fruit production.
An attempt was made to solve these problems by screening drought and salt tolerance genes in the tolerant cultivars, ‘Niao’ and a plum-mume parental line ‘PM-1-1’, and introducing them into ‘Nanko’. Potted plants of the three cultivars grown in a greenhouse were unstressed by watering (control), stressed by withholding watering (drought stress) or by applying 100 mM NaCl solution (salt stress). Total RNA samples were extracted from young leaves of each cultivar 0, 3 and 7 days after the start of stress treatments.
Differential display was performed to compare overall differences in gene expression among the treatments and cultivars.
The total RNA samples were reverse-transcribed using T(12)VC and T(12)VG primers, followed by PCR amplification using two anchoring primers, T(12)VC and T(12)VG, and one hundred arbitrary 10-mer primers (OPBD-OPBH, Operon, USA). After screening 200 primer combinations, seemingly, 43 cDNA fragments were differentially expressed in response to drought stress, whereas 15 fragments were induced under salt stress.
BLAST analysis revealed that 29 fragments of cDNAs with known function had high homology to 9 known stress-related genes: abscisic stress ripening-like protein mRNA of Prunus persica, ripening protein mRNA of Fragaria × ananassa, auxin-repressed protein mRNA of Prunus ameniaca, auxin-regulated protein GH3 gene of Glycine max, seed imbibition (dehydration) protein mRNA of Persa Americana, heat shock 70 protein mRNA of Spinacia oleracea, histone H2A (water-deficit stress) of Oryza sativa, extensin cDNA of Solanum tuberosum and geranylgeranyl hydrogenase mRNA of Triticum aestivum.
Recent warm, dry weather and salt-contaminated irrigation have resulted in poor field establishment of stocks and decreased fruit production.
An attempt was made to solve these problems by screening drought and salt tolerance genes in the tolerant cultivars, ‘Niao’ and a plum-mume parental line ‘PM-1-1’, and introducing them into ‘Nanko’. Potted plants of the three cultivars grown in a greenhouse were unstressed by watering (control), stressed by withholding watering (drought stress) or by applying 100 mM NaCl solution (salt stress). Total RNA samples were extracted from young leaves of each cultivar 0, 3 and 7 days after the start of stress treatments.
Differential display was performed to compare overall differences in gene expression among the treatments and cultivars.
The total RNA samples were reverse-transcribed using T(12)VC and T(12)VG primers, followed by PCR amplification using two anchoring primers, T(12)VC and T(12)VG, and one hundred arbitrary 10-mer primers (OPBD-OPBH, Operon, USA). After screening 200 primer combinations, seemingly, 43 cDNA fragments were differentially expressed in response to drought stress, whereas 15 fragments were induced under salt stress.
BLAST analysis revealed that 29 fragments of cDNAs with known function had high homology to 9 known stress-related genes: abscisic stress ripening-like protein mRNA of Prunus persica, ripening protein mRNA of Fragaria × ananassa, auxin-repressed protein mRNA of Prunus ameniaca, auxin-regulated protein GH3 gene of Glycine max, seed imbibition (dehydration) protein mRNA of Persa Americana, heat shock 70 protein mRNA of Spinacia oleracea, histone H2A (water-deficit stress) of Oryza sativa, extensin cDNA of Solanum tuberosum and geranylgeranyl hydrogenase mRNA of Triticum aestivum.
Authors
S. Wongchaochant, K. Negoro, M. Kou, H. Hanada
Keywords
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