Study on factors influencing transformation efficiency in Pinus massoniana using Agrobacterium tumefaciens

Maleki, Samaneh Sadat; Mohammadi, Kourosh; Ji, K.

doi:10.1007/s11240-018-1388-7

Cited by 23 publications

(17 citation statements)

References 32 publications

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“…Masson pine ( Pinus massoniana ) is an economically and ecologically important evergreen conifer in southern China and is one of the most important forest tree species 18 , 19 . It is widely used in the production of solid wood and resin 20 .…”

Section: Introductionmentioning

confidence: 99%

Identification, classification, and characterization of AP2/ERF superfamily genes in Masson pine (Pinus massoniana Lamb.)

Zhu

Chen

Zhang

et al. 2021

Sci Rep

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Transcription factors (TFs) play crucial regulatory roles in controlling the expression of the target genes in plants. APETALA2/Ethylene-responsive factors (AP2/ERF) are part of a large superfamily of plant-specific TFs whose members are involved in the control of plant metabolism, development and responses to various biotic and abiotic stresses. However, the AP2/ERF superfamily has not been identified systematically in Masson pine (Pinus massoniana), which is one of the most important conifer in southern China. Therefore, we performed systematic identification of the AP2/ERF superfamily using transcriptome sequencing data from Masson pine. In the current study, we obtained 88 members of the AP2/ERF superfamily. All PmAP2/ERF members could be classified into 3 main families, AP2 (7 members), RAV (7 members), ERF (73 members) families, and a soloist protein. Subcellular localization assays suggested that two members of PmAP2/ERF were nuclear proteins. Based on pine wood nematode (PWN) inoculated transcriptome and qPCR analysis, we found that many members of PmAP2/ERF could respond to PWN inoculation and PWN related treatment conditions in vitro. In general, members of the AP2/ERF superfamily play an important role in the response of Masson pine responds to PWN. Furthermore, the roles of the AP2/ERF superfamily in other physiological activities of Masson pine remain to be further studied.

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Section: Introductionmentioning

confidence: 99%

Identification, classification, and characterization of AP2/ERF superfamily genes in Masson pine (Pinus massoniana Lamb.)

Zhu

Chen

Zhang

et al. 2021

Sci Rep

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“…Pinus massoniana (Masson pine) is an economically and ecologically important evergreen conifer in southern China and is one of the most significant forest tree species [22,23]. It is widely used in the production of fiber and solid wood [24,25].…”

Section: Introductionmentioning

confidence: 99%

Selection of Suitable Reference Genes in Pinus massoniana Lamb. Under Different Abiotic Stresses for qPCR Normalization

Zhu

et al. 2019

Forests

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The normalization of data by choosing suitable reference genes is fundamental for obtaining accurate and reliable results in quantitative real-time polymerase chain reaction (qPCR) analyses. In this study, the expression stability of 12 candidate reference genes of Pinus massoniana under different abiotic stresses was evaluated using four statistical algorithms: geNorm, NormFinder, BestKeeper, and RefFinder. The results indicate that the following genes could be used as reference genes under different treatments: Actin 2 (ACT2) and F-box family gene (F-box) for salinity treatment, cyclophilin (CYP) and alpha-tubulin (TUA) for ABA treatment, actin 7 (ACT7) and CYP for drought treatment, actin 1 (ACT1) and ACT7 for cold treatment, ACT1 and CYP for heat treatment, and TUA and ACT2 for the “Total” group. To validate the suitability of the selected reference genes in this study, the Short-Root protein (SHR), Alpha-pinene synthase (APS), and Pyrabactin resistance-like protein (PYL) gene expression patterns were analyzed. The expression patterns had significant biases when the most unstable reference genes were used for normalization, compared with when the optimum reference gene or gene combinations were used for normalization. These results will be beneficial for further studies on gene transcription in early-stage, unlignified seedlings of P. massoniana.

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“…[38]. The method has since been extensively used in other conifer species, including Norway spruce, Loblolly pine [15], White pine [8], P. massoniana [11], Slash pine [13], P. patula [39], and P. radiata [40]. Use of embryos, cotyledons, hypocotyls, and female gametophytes as explants reportedly induces non-embryogenic or embryogenic calli and adventitious buds [41][42][43][44].…”

Section: Discussionmentioning

confidence: 99%

“…This method is one such versatile simple, rapid and transient expression of target genes can be detected within a few hours [8,16,27]. However, the average transformation efficiency is affected by the Agrobacterium strain [13], explant type [28], Agrobacterium density [29], acetosyringone (AS) concentration [30], and time period [11]. In this study, we obtained a hypocotyl-derived callus from P. tabuliformis seedlings, which promoted transient transformation.…”

Section: Open Accessmentioning

confidence: 99%

“…The Agrobacterium-mediated method, including stable transformation and transient gene expression, has been functional in number of plant species [6][7][8][9][10]. Although stable and resilient transformation protocols has been successfully carried out in a few conifer species, such as P. massoniana [11], Korean fir [12], Slash pine [13], Maritime pine [14], Norway spruce, and Loblolly pine [15]. However, there is still major technical obstacle…”

Section: Introductionmentioning

confidence: 99%

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An efficient system for Agrobacterium-mediated transient transformation in Pinus tabuliformis

Liu

et al. 2020

Plant Methods

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Background: Functional genomic studies using genetics approaches of conifers are hampered by the complex and enormous genome, long vegetative growth period, and exertion in genetic transformation. Thus, the research carried out on gene function in Pinus tabuliformis is typically performed by heterologous expression based on the model plant Arabidopsis. However, due to the evolutionary and vast diversification from non-flowering (gymnosperms) to flowering (angiosperms) plants, several key differences may alter the underlying genetic concerns and the analysis of variants. Therefore, it is essential to develop an efficient genetic transformation and gene function identification protocol for P. tabuliformis. Results: In the present study we established a highly efficient transgene Agrobacterium-mediated transient expression system for P. tabuliformis. Using a β-glucuronidase gene (GUS) as a reporter gene expression, the highest transformation efficiency (70.1%) was obtained by co-cultivation with Agrobacterium strain GV3101 at an optical density at 600 nm of 0.8, with 150 μM acetosyringone for 30 min followed by 3 days in the dark at 23 ± 1 °C. This protocol would be applied to other conifers; GUS staining was observed 24 h post-infection. Conclusions: We report a simple, fast, and resilient system for transient Agrobacterium-mediated transformation high-level expression of target genes in P. tabuliformis, which will also improve transformation efficiency in other conifer species.

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Study on factors influencing transformation efficiency in Pinus massoniana using Agrobacterium tumefaciens

Cited by 23 publications

References 32 publications

Identification, classification, and characterization of AP2/ERF superfamily genes in Masson pine (Pinus massoniana Lamb.)

Identification, classification, and characterization of AP2/ERF superfamily genes in Masson pine (Pinus massoniana Lamb.)

Selection of Suitable Reference Genes in Pinus massoniana Lamb. Under Different Abiotic Stresses for qPCR Normalization

An efficient system for Agrobacterium-mediated transient transformation in Pinus tabuliformis

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