SummaryThe genes that encode the ethylene biosynthesis enzyme 1‐aminocyclopropane‐1‐carboxylate oxidase (ACO) are thought to be involved in flower senescence. Hence, we investigated whether the transcript levels of PhACO genes (PhACO1, PhACO3 and PhACO4) in Petunia cv. Mirage Rose are associated with ethylene production at different flowering stages. High transcript levels were detected in the late flowering stage and linked to high ethylene levels. PhACO1 was subsequently edited using the CRISPR/Cas9 system, and its role in ethylene production was investigated. PhACO1‐edited T0 mutant lines, regardless of mutant type (homozygous or monoallelic), exhibited significantly reduced ethylene production and enhanced flower longevity compared with wild‐type. Flower longevity and the reduction in ethylene production were observed to be stronger in homozygous plants than in their monoallelic counterparts. Additionally, the transmission of the edited gene to the T1 (lines 6 and 36) generation was also confirmed, with the results for flower longevity and ethylene production proving to be identical to those of the T0 mutant lines. Overall, this study increases the understanding of the role of PhACO1 in petunia flower longevity and also points to the CRISPR/Cas9 system being a powerful tool in the improvement of floricultural quality.
Resveratrol Synthase (RS) is one of the key enzymes in resveratrol biosynthesis, which catalyzes one molecule of coumaroyl CoA and three molecules of malonyl CoA to form one molecule of resveratrol. In this study, the genetic transformation system for badila, a chewing cane variety, was established. The optimized media for regeneration were as follows: induction medium was MS+2,4-D 2mg.L -1 ; differentiation medium was MS+NAA 0.2 mg.L -1 +BA 2 mg.L -1 ; rooting medium was 1/2 MS+NAA 2 mg.L -1 . The addition of 0.5 % activated carbon in the differentiation and rooting medium could decrease the phenol poison and promote rooting. At the same time, the optimal concentration of G418 in subculture and differentiation period was determined as: 30 mgL -1 for differentiation and 25 mgL -1 for rooting. RS gene was transferred into badila via gene gun bombardment through the constructed plant expression vector pBIL-RS. PCR detection and southern blot analysis showed that seven putative transgenic plants had been obtained. Therefore, the genetic transformation system of badila had been elementarily established and proved to be efficient.
Although a large number of AroA enzymes (EPSPS: 5-enopyruvylshikimate-3-phosphate synthase) have been identified, cloned, and tested for glyphosate resistance, only two AroA variants, derived from Agrobacterium tumefaciens strain CP4 and Zea mays, have been utilized to produce the commercial glyphosate-resistant crops. Here, we have used a PCR-based twostep DNA synthesis method to synthesize an aroA gene (aroAA. metalliredigens) from Alkaliphilus metalliredigens, encoding a new EPSPS. Furthermore, transgenic Arabidopsis with the new aroAA. metalliredigens gene was obtained to confirm the potential of the novel aroA gene in developing glyphosate-resistant crops.
This study was conducted to examine the role of the transcription factors (TFs), RsMYB1 and mPAP1 together with B-Peru (mPAP1 + B-Peru), in regulating anthocyanin biosynthesis in the ornamental torenia (Torenia fournieri) cultivar Kauai Rose using Agrobacterium-mediated transformation. Expression levels of RsMYB1 were the highest in the lines RS5 and RS3, followed by RS4, RS2, and RS1, while transcript levels of mPAP1 + B-Peru increased in the order of PB-6 > PB-5 > PB-7 > PB-8 > PB-2. Moreover, transcript levels of the anthocyanin structural genes in transgenic lines were significantly higher than those in wild-type (WT) plants. Anthocyanin structural gene expression was specifically altered by TF overexpression: the highest expression of anthocyanidin synthase (ANS) was observed in transgenic lines with RsMYB1, while expression of dihydroflavonol-4-reductase (DFR) was the highest in lines with mPAP1 + B-Peru. We expect that enhanced expression of these anthocyanin structural genes will improve anthocyanin content in the flowers of transgenic torenia. Moreover, these results indicate that RsMYB1 and mPAP1 + B-Peru can be exploited as anthocyanin regulatory TFs to enhance anthocyanin content in other horticultural plants.
KeywordsFlower color • Genetic transformation • Ornamental plant • Quantitative real-time polymerase chain reaction (qRT-PCR) • Anthocyanin • RsMYB1 • mPAP1 • B-Peru Aung Htay Naing and Xu Junping equally contributed as first authors.
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