Cloning of RhRNF185-like Gene from Rose and Its Effect on Petal Senescence

doi:10.16420/j.issn.0513-353x.2023-0373

Acta Horticulturae Sinica ›› 2024, Vol. 51 ›› Issue (5): 1047-1055.doi: 10.16420/j.issn.0513-353x.2023-0373

• Genetic & Breeding·Germplasm Resources·Molecular Biology • Previous Articles Next Articles

Cloning of RhRNF185-like Gene from Rose and Its Effect on Petal Senescence

WANG Wenyu, WANG Jiayin, DU Tingting, ZHANG Jingjing, ZHANG Chao, XIN Cuihua, GUO Jiangbo, PEI Haixia^*()

College of Life Science and Technology，Inner Mongolia University of Science and Technology，Baotou，Inner Mongolia 014010，China

Received:2023-05-30 Revised:2023-09-10 Online:2024-05-25 Published:2024-05-29

Abstract

Abstract:

In order to explore the senescence mechanism of rose petals and provide gene reserve for the creation of anti-senescence rose varieties through molecular breeding and other means，a RhRNF185-like gene belonging to E3 ubiquitin ligase RING domain family were screened by transcriptome sequencing using cut rose‘Carola’as the test material in this study. The results of qRT-PCR showed that the expression level of RhRNF185-like in the rose petals was significantly increased in the middle and late stage of rose flower opening，indicating that RhRNF185-like expression might affect the senescence of rose petals. Transient overexpression of RhRNF185-like in tobacco mesophyll cells showed that the GFP signal of RhRNF185-like coincided with the nuclear signal of DAPI through Confocal microscope，indicating that RhRNF185-like was located in the nucleus. Virus-induced gene silencing technique（VIGS）was used to silence RhRNF185-like gene in rose petals. It was found that the senescence degree of petals with RhRNF185-like gene silencing was significantly greater than that of TRV control group. In addition，compared with TRV control group，the relative expression level of senescence marker gene RhSAG12，ion permeability and fading rate of rose petals were significantly increased in the RhRNF185-like gene silencing group，indicating that silencing RhRNF185-like promoted the senescence of rose petals. These results indicated that RhRNF185-like affected the senescence of rose petals in an antagonistic regulatory manner.

Key words: rose, petal, senescence, RhRNF185-like

WANG Wenyu, WANG Jiayin, DU Tingting, ZHANG Jingjing, ZHANG Chao, XIN Cuihua, GUO Jiangbo, PEI Haixia. Cloning of RhRNF185-like Gene from Rose and Its Effect on Petal Senescence[J]. Acta Horticulturae Sinica, 2024, 51(5): 1047-1055.

Figures/Tables 7

Table 1 The sequences of primers used for experiment

引物名称 Primer name	引物序列（5′-3′） Primer sequence
RhUBI2-qPCR	F：GCCCTGGTGCGTTCCCAACTG；R：CCTGCGTGTCTGTCCGCATTG
RhRNF185-like-qPCR	F：CAATGACGCAGGGGATTTCG；R：ATGTGAGTGATGGTGGAGCC
RhRNF185-like-VIGS	F：GTTTTGGGGAATCAACGAGC；R：TCAGCGCAGCAATCACAAAT
RhRNF185-like-ORF	F：ATGGCAAGTGGTTTTGGGGAAT；R：CTAACAAAAAATCAGCGCAGC
SAG12-qPCR	F：AAGACCAATCCAAAAGCAAC；R：AATCAAAACCACCTCCTTCA

Fig. 1 Relative expression of RhRNF185-like in different opening grades of Chinese rose * P < 0.05，** P < 0.01.

Fig. 2 ORF cloning of RhRNF185-like（A）and screening and identification of silencing vector pYY13 and overexpression localization vector pJG186（B，C）

Fig. 3 Phylogenetic tree analysis of RhRNF185-like FV：Fragaria vesca subsp.；Md：Malus × domestica；Mn：Morus notabilis；Pp：Prunus persica；Pm：Prunus mume；Me：Manihot esculenta.

Fig. 4 RhRNF185-like subcellular localization

Fig. 5 Phenotype of rose petal disc RhRNF185-like silencing and relative expression of this gene and senescence marker gene RhSAG12 ** P < 0.01.

Fig. 6 Ion permeability and color fading rate of rose petal disc RhRNF185-like after silencing * P < 0.05，** P < 0.01.

References 27

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Cloning of RhRNF185-like Gene from Rose and Its Effect on Petal Senescence

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