Phenotypic Characterization and RT-qPCR Analysis of Flower Development in F<sub>1</sub> Transgenics of Chrysanthemum × grandiflorum
Gene silencing is the epigenetic regulation of any gene in order to prevent gene expression at the transcription or translation levels. Among various gene silencing techniques, RNA silencing (RNAi) is notable gene regulation technique that involves sequence-specific targeting and RNA degradation. Ho...
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MDPI AG
2021-08-01
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author | Saba Haider Muhammad Ajmal Bashir Umer Habib Yike Gao Muhammad Rashid Shaheen Rashid Hussain Fan Min |
author_facet | Saba Haider Muhammad Ajmal Bashir Umer Habib Yike Gao Muhammad Rashid Shaheen Rashid Hussain Fan Min |
author_sort | Saba Haider |
collection | DOAJ |
description | Gene silencing is the epigenetic regulation of any gene in order to prevent gene expression at the transcription or translation levels. Among various gene silencing techniques, RNA silencing (RNAi) is notable gene regulation technique that involves sequence-specific targeting and RNA degradation. However, the effectiveness of transgene-induced RNAi in F<sub>1</sub> generation of chrysanthemum has not been studied yet. In the current study, we used RNAi-constructed <i>CmTFL1</i> (white-flowered) and <i>CmSVP</i> overexpressed (yellow flowered) transgenic plants of previously conducted two studies for our experiment. Cross hybridization was performed between these intergeneric transgenic and non-transgenic plants of the winter-growing chrysanthemum selection “37” (light pink flowered). The transgene <i>CmSVP</i> was confirmed in F<sub>1</sub> hybrids by RT-PCR analysis, whereas hybrids of <i>CmTFL1</i> parental plants were non-transgenic. Besides this, quantitative real-time PCR (qPCR) was used to explain the molecular mechanism of flower development using reference genes. Intergeneric and interspecific hybrids produced different colored flowers unlike their respective parents. These results suggest that generic traits of <i>CmSVP</i> overexpressed plants can be transferred into F<sub>1</sub> generations when crossed with mutant plants. This study will aid in understanding the breeding phenomenon among intergeneric hybrids of chrysanthemum plants at an in vivo level, and such transgenics will also be more suitable for sustainable flower yield under a low-light production system. |
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spelling | doaj.art-d90592fb62924d9c82660712e212e4ed2023-11-22T09:20:03ZengMDPI AGPlants2223-77472021-08-01108168110.3390/plants10081681Phenotypic Characterization and RT-qPCR Analysis of Flower Development in F<sub>1</sub> Transgenics of Chrysanthemum × grandiflorumSaba Haider0Muhammad Ajmal Bashir1Umer Habib2Yike Gao3Muhammad Rashid Shaheen4Rashid Hussain5Fan Min6National Flower Engineering Research Centre, Beijing Key Laboratory of Ornamental Plants Germplasm Innovation and Molecular Breeding, College of Landscape Architecture, Beijing Forestry University, Beijing 100083, ChinaLaboratory of Plant Biotechnology and In Vitro Tissue Culture, Department of Agriculture and Forest Sciences (DAFNE), University of Tuscia, 01100 Viterbo, ItalyDepartment of Horticulture, PMAS-Arid Agriculture University, Rawalpindi 46300, PakistanNational Flower Engineering Research Centre, Beijing Key Laboratory of Ornamental Plants Germplasm Innovation and Molecular Breeding, College of Landscape Architecture, Beijing Forestry University, Beijing 100083, ChinaDepartment of Horticultural Sciences, Faculty of Agriculture and Environment, The Islamia University of Bahawalpur, Punjab 63100, PakistanDepartment of Horticultural Sciences, Faculty of Agriculture and Environment, The Islamia University of Bahawalpur, Punjab 63100, PakistanNational Flower Engineering Research Centre, Beijing Key Laboratory of Ornamental Plants Germplasm Innovation and Molecular Breeding, College of Landscape Architecture, Beijing Forestry University, Beijing 100083, ChinaGene silencing is the epigenetic regulation of any gene in order to prevent gene expression at the transcription or translation levels. Among various gene silencing techniques, RNA silencing (RNAi) is notable gene regulation technique that involves sequence-specific targeting and RNA degradation. However, the effectiveness of transgene-induced RNAi in F<sub>1</sub> generation of chrysanthemum has not been studied yet. In the current study, we used RNAi-constructed <i>CmTFL1</i> (white-flowered) and <i>CmSVP</i> overexpressed (yellow flowered) transgenic plants of previously conducted two studies for our experiment. Cross hybridization was performed between these intergeneric transgenic and non-transgenic plants of the winter-growing chrysanthemum selection “37” (light pink flowered). The transgene <i>CmSVP</i> was confirmed in F<sub>1</sub> hybrids by RT-PCR analysis, whereas hybrids of <i>CmTFL1</i> parental plants were non-transgenic. Besides this, quantitative real-time PCR (qPCR) was used to explain the molecular mechanism of flower development using reference genes. Intergeneric and interspecific hybrids produced different colored flowers unlike their respective parents. These results suggest that generic traits of <i>CmSVP</i> overexpressed plants can be transferred into F<sub>1</sub> generations when crossed with mutant plants. This study will aid in understanding the breeding phenomenon among intergeneric hybrids of chrysanthemum plants at an in vivo level, and such transgenics will also be more suitable for sustainable flower yield under a low-light production system.https://www.mdpi.com/2223-7747/10/8/1681RNA silencingcross breedinghybrid identificationquantitative real time PCR |
spellingShingle | Saba Haider Muhammad Ajmal Bashir Umer Habib Yike Gao Muhammad Rashid Shaheen Rashid Hussain Fan Min Phenotypic Characterization and RT-qPCR Analysis of Flower Development in F<sub>1</sub> Transgenics of Chrysanthemum × grandiflorum Plants RNA silencing cross breeding hybrid identification quantitative real time PCR |
title | Phenotypic Characterization and RT-qPCR Analysis of Flower Development in F<sub>1</sub> Transgenics of Chrysanthemum × grandiflorum |
title_full | Phenotypic Characterization and RT-qPCR Analysis of Flower Development in F<sub>1</sub> Transgenics of Chrysanthemum × grandiflorum |
title_fullStr | Phenotypic Characterization and RT-qPCR Analysis of Flower Development in F<sub>1</sub> Transgenics of Chrysanthemum × grandiflorum |
title_full_unstemmed | Phenotypic Characterization and RT-qPCR Analysis of Flower Development in F<sub>1</sub> Transgenics of Chrysanthemum × grandiflorum |
title_short | Phenotypic Characterization and RT-qPCR Analysis of Flower Development in F<sub>1</sub> Transgenics of Chrysanthemum × grandiflorum |
title_sort | phenotypic characterization and rt qpcr analysis of flower development in f sub 1 sub transgenics of chrysanthemum grandiflorum |
topic | RNA silencing cross breeding hybrid identification quantitative real time PCR |
url | https://www.mdpi.com/2223-7747/10/8/1681 |
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