‘鲁星’桃中10个MADS-box基因克隆和表达分析

doi:10.3864/j.issn.0578-1752.2016.23.012

Abstract

Abstract: 【Objective】The aim of this study is to characterize the novel peach (Prunus persica var. nectarina ‘Luxing’) MADS-box genes (PpMADSs) involved in regulation of vegetative and reproductive growth. The transcriptional levels of PpMADSs in different tissues were determined to provide a basis for studying the related function of PpMADSs in flower development, fruit development and ripening.【Method】The full-length cDNA sequences of PpMADSs form ‘Luxing’ peach were isolated by homologous alignment and RT-PCR confirmation, the obtained cDNA sequences and the deduced amino acid sequences were analyzed with bioinformatics methods; the expression levels of PpMADSs were detected in stem, leaf, sepal, ovary, stamen, petal, 7 stages of flower development and 5 stages of fruit development using RT-PCR.【Result】The sequencing results showed that ten cDNAs (designated as PpMADS11, 12, 19, 20, 21, 22, 28, 29, 30 and 31; GenBank accession No. KU559577, KU559578, KU559585, KU559586, KU559587, KU559588, KU559594, KU559595, KU559596 and KU559597) contained open reading frame (ORF) of 522, 279, 1 065, 828, 723, 600, 636, 534, 750 and 480 bp, respectively. The results of phylogenetic analysis revealed that PpMADS11, 12, and 19 belong to AP3, AGL17 and MIKC* subgroups, respectively; and PpMADS20, 21 and 22 belong to Mα group; PpMADS28, 29, 30 and 31 belong to Mγ group. The results of subcellular localization prediction showed that all PpMADS proteins were located in the nucleus. The results of promoter analysis indicated that there were multiple putative cis-acting elements involved in light responsiveness, defense and stress responsiveness, MYB binding site was involved in drought-inducibility, heat stress responsiveness, low-temperature responsiveness, fungal elicitor responsive element, wound-responsive element, anaerobic induction element, gibberellin-responsive element, auxin-responsive element, MeJA-responsiveness, abscisic acid responsiveness, salicylic acid responsiveness and ethylene-responsive element. Semi RT-PCR and qRT-PCR results showed that PpMADS11was expressed in stem, leaf, sepal, ovary, stamen, petal and during flower and fruit development. PpMADS12 was expressed in stem, leaf, sepal, ovary, stamen, petal and during flower development. PpMADS19 was expressed in sepal, stamen, petal and during flower development ( except bud stage). All members in Mα and Mγ groups were expressed in stem, leaf, sepal, ovary, stamen, petal and during flower development, some members were expressed during fruit development.【Conclusion】These results indicated that ten PpMADS genes have crucial regulatory roles in ‘Luxing’ peach vegetative growth, flower and fruit development processes.

Key words: ‘Luxing&rsquo, peach, MADS-box, transcription factor, gene cloning, expression analysis

LI Hui-feng, JIA Hou-zhen, DONG Qing-long, RAN Kun, WANG Hong-wei. Cloning and Expression Analysis of Ten MADS-box Genes in Peach (Prunus persica var. nectarina ‘Luxing’)[J].Scientia Agricultura Sinica, 2016, 49(23): 4593-4605.

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Cloning and Expression Analysis of Ten MADS-box Genes in Peach (Prunus persica var. nectarina ‘Luxing’)

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