Characterization of miR061 and its target genes in grapevine responding to exogenous gibberellic acid

Wang, Mengqi; Sun, Xiaowen; Wang, Chen; Cui, Liwen; Lide, Chen; Zhang, Chaobo; Shangguan, Lingfei; Fang, Jinggui

doi:10.1007/s10142-017-0554-z

Cited by 15 publications

(17 citation statements)

References 59 publications

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“…Therefore, GA is recognized as a key phytohormone regulator for seed embryo development and stone-hardening in grape. Previous studies showed that several VvmiRNAs were differentially expressed during grape berry development in response to GA application (Ren et al 2014, Han et al 2014, Wang et al 2017. However, stone-hardening stage-speci c VvmiRNAs and their regulatory networks during grape berry development remain imperative.…”

Section: Discussionmentioning

confidence: 99%

Study on miRNAs-Mediated Seed and Stone-Hardening Regulatory Networks and the Mechanism of miRNAs’ Manipulating Gibberellin-Induced Seedless Berries in Grapevine (Vitis vinifera L.)

Wang

et al. 2020

Preprint

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A significant body of evidence supports the important roles of miRNAs in grape berry developments. However, their specific molecular functions during stone-hardening stage development remain unclear. Here, a total of 161 conserved and 85 species-specific miRNAs/miRNAs*(precursor) were identified in grape stone hardening stage berries using Solexa sequencing. Out of them, 30 VvmiRNAs were tissue-specific and identified as stone-hardening related to VvmiRNAs, whereas 52 exhibited differential expression profiles during berry development, potentially participating in modulation of development, and qRT-PCR analysis verified their expression patterns. Interestingly, high SNP variations in VvmiRNA sequences might result into generation of new VvmiRNA family members like VvmiR168, VvmiR479, VvmiR3636 families and so on. Through GO and KEGG pathway analyses, we revealed that 13 VvmiRNAs involved in the regulation of embryo development, 11 in lignin and cellulose biosynthesis, and 28 in the modulation of hormone signaling, sugar and proline metabolism. Furthermore, the target genes for novel VvmiRNAs related to berry development were validated using RLM-RACE and PPM-RACE methods, and it was revealed their cleavage sites mainly happened at the 9th-11th sites from the 5’ ends of miRNAs at their binding regions. The potential roles of these VvmiRNAs in gibberellins repressing grape stone-hardening and embryo development by potentially inducing the expression of VvmiR31-3p and VvmiR8-5p to increase the cleavage product accumulation levels of corresponding target genes like lignin biosynthesis genes, CAFFEOYL COENZYME A-3-O-METHYLTRANSFERASE (VvCCoAOMT) and DDB1-CUL4 ASSOCIATED FACTOR1 (VvDCAF1), as a potential key molecular mechanism involved in GA-induced grape seedless berry development. Based on the characterization of stone-hardening stage related to VvmiRNAs, a schematic model of miRNA-mediated grape seed and stone-hardening development was proposed in this work. This is the first report about the regulatory role of VvmiRNAs in the regulation of stone-hardening stage of grape berries, which provides valuable genetic information for the breeding of seedless grape varieties.

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Section: Discussionmentioning

confidence: 99%

Study on miRNAs-Mediated Seed and Stone-Hardening Regulatory Networks and the Mechanism of miRNAs’ Manipulating Gibberellin-Induced Seedless Berries in Grapevine (Vitis vinifera L.)

Wang

et al. 2020

Preprint

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“…The effects of exogenous GA application on the induction of flower development, seedless cultivars, berry enlargement and fruit ripening of grapevine have recently been reported [18, 20]. However, the molecular mechanisms by which GA-signaling mediates flower and fruit set initiation in grapevines are still not fully understood.…”

Section: Discussionmentioning

confidence: 99%

“…Phytohormones, such as GA, are known to be essential for the regulation of grape flower development, berry expansion, ripening, and seedless berry induction, and the exogenous application of GA can trigger fruit parthenocarpy and early ripening in various grapevine cultivars [15, 18, 19]. Several reports demonstrated that different miRNA families (miR061, miR156, miR159, miR160, miR164, miR167, miR172, and miR319) and their target genes are implicated in GA signal during floral transition and fruit set development [8, 9, 20]. For instance, Arabidopsis AtmiR159 acts as a homeostatic regulator of GA signaling to direct the cleavage of genes encoding AtGAMYB proteins, causing a defect in anthers development and a subsequent reduction in floral fertility [21].…”

Section: Introductionmentioning

confidence: 99%

VvmiR160s/VvARFs interaction and their spatio-temporal expression/cleavage products during GA-induced grape parthenocarpy

et al. 2019

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BackgroundGrape (Vitis vinifera) is highly sensitive to gibberellin (GA), which effectively induce grape parthenocarpy. Studies showed that miR160s and their target AUXIN RESPONSIVE FACTOR (ARF) responding hormones are indispensable for various aspects of plant growth and development, but their functions in GA-induced grape parthenocarpy remain elusive.ResultsIn this study, the morphological changes during flower development in response to GA treatments were examined in the ‘Rosario Bianco’ cultivar. The precise sequences of VvmiR160a/b/c/d/e and their VvARF10/16/17 target genes were cloned, sequenced and characterized. The phylogenetic relationship and intron-exon structure of VvARFs and other ARF family members derived from different species were investigated. All VvmiR160s (except VvmiR160b) and VvARF10/16/17 had the common cis-elements responsive to GA, which support their function in GA-mediated grape parthenocarpy. The cleavage role of VvmiR160s-mediated VvARF10/16/17 was verified in grape flowers. Moreover, spatio-temporal expression analysis demonstrated that among VvmiR160 family, VvmiR160a/b/c highly expressed at late stage of flower/berry development, while VvARF10/16/17showed a reverse expression trend. Interestingly, GA exhibited a long-term effect through inducing the expression of VvmiR160a/b/c/e to increase their cleavage product accumulations from 5 to 9 days after treatment, but GA enhanced the expressions of VvARF10/16/17 only at short term. Pearson correlation analysis based on expression data revealed a negative correlation between VvmiR160a/b/c and VvARF10/16/17 in flowers not berries during GA-induced grape parthenocarpy.ConclusionsThis work demonstrated that the negative regulation of VvARF10/16/17 expression by VvmiR160a/b/c as key regulatory factors is critical for GA-mediated grape parthenocarpy, and provide significant implications for molecular breeding of high-quality seedless berry.Electronic supplementary materialThe online version of this article (10.1186/s12870-019-1719-9) contains supplementary material, which is available to authorized users.

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“…RNA was isolated from grapevine plant tissue using standard molecular biology protocols and following manufacturer’s instructions, as previously described [ 5 ].…”

Section: Methodsmentioning

confidence: 99%

“…In plants, miRNAs play important roles in a number of physiological processes, such as growth development and biotic and abiotic stress responses [ 1 – 5 ]; they can regulate the expression levels of their target genes by cleavage. Other miRNAs emerged as promising approaches for plant breeding applications, such as crop improvement strategies [ 6 , 7 ].…”

Section: Introductionmentioning

confidence: 99%

Characterization of the regulation mechanism of grapevine microRNA172 family members during flower development

et al. 2020

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Background Grapevine ( Vitis vinifera L . ), which has important nutritional values and health benefits, is one of the most economically important fruit crops cultivated worldwide. Several studies showed a large number of microRNAs (VvmiRNAs) involved in the modulation of grape growth and development, and many VvmiRNA families have multiple members. However, the way by which various members from the same miRNA family work is unclear, particularly in grapes. Results In this study, an important conserved VvmiR172 family (VvmiR172s) and their targets were set as a good example for elucidating the interaction degree, mechanism, and spatio-temporal traits of diverse members from the same miRNA family. miR-RACE and Stem-loop RT-PCR were used to identify the spatio-temporal expressions of various members of VvmiR172s; together with RLM-RACE, PPM-RACE, Western blot, transgenic technologies, their interaction degree, and regulation mechanism were further validated. The expression of VvmiR172c was significantly higher than that of VvmiR172a, b, and d and showed a positive correlation with the abundance of VvAP2 cleavage products. These findings indicated that VvmiR172c might be one of the main action factors of the VvmiR172 family in flower development. The ability of VvmiR172c to cleave target genes differed due to divergence in complementary degree with VvAP2 and expression levels of various members. In VvmiR172 transgenic lines, we observed that 35S::VvmiR172c resulted in the earliest and abundant flowering, indicating the strong function of VvmiR172c. In contrast, the non-significant phenotypic changes were detected in the VvAP2 transgenic lines. The qRT-PCR and Western bolt results demonstrated that VvmiR172c plays a major role in targeting VvAP2 . Conclusions VvmiR172 up-regulated the expression of NtFT and decreased the expression of NtFLC . The up/down regulation of VvmiR172c was the most pronounced. The functions of four VvmiR172 members in grape differed, and miR172c had the strongest regulation on AP2 .

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Characterization of miR061 and its target genes in grapevine responding to exogenous gibberellic acid

Cited by 15 publications

References 59 publications

Study on miRNAs-Mediated Seed and Stone-Hardening Regulatory Networks and the Mechanism of miRNAs’ Manipulating Gibberellin-Induced Seedless Berries in Grapevine (Vitis vinifera L.)

Study on miRNAs-Mediated Seed and Stone-Hardening Regulatory Networks and the Mechanism of miRNAs’ Manipulating Gibberellin-Induced Seedless Berries in Grapevine (Vitis vinifera L.)

VvmiR160s/VvARFs interaction and their spatio-temporal expression/cleavage products during GA-induced grape parthenocarpy

Characterization of the regulation mechanism of grapevine microRNA172 family members during flower development

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