Phytase expression in transgenic soybeans: stable transformation with a vector-less construct

Xiao, Guohui; Wang, Guo Kun; Su, Qiao; Wang, Yan; Liu, An

doi:10.1007/s10529-007-9439-x

Cited by 33 publications

(21 citation statements)

References 16 publications

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“…The linear minimal gus reporter gene cassette was generated by polymerase chain reaction (PCR) using pBI121 (Clontech, USA) as a template and the primers PT1 (5′-tggcaggatatattgtggtgtaaactgcctgcaggtccccagatt agcctt-3′) and PT2 (5′-gtttacccgccaatatatcctgtcaccgatct agtaacatatgagacaccgc-3′), according to Gao et al (2007). The relative positions of all primers on the minimal gus reporter gene cassette are shown in Fig.1.…”

Section: Preparation Of Dna Cassettes For Ovary Transformationmentioning

confidence: 99%

“…A shortened pathway, by partial ovary cutting, facilitated the entry of DNA molecules into the embryo sac ), but also resulted in reduced pod-bearing rates (Gao et al, 2007). In this study, we compared the effects of four types of style cutting on transformation efficiency in detail.…”

Section: Effects Of Different Style Cutting Treatments On Soybean Ovamentioning

confidence: 99%

“…Recently, direct transformation of a minimal linear gene cassette by stigma or ovary-drip without using bacteria has been reported in corn and soybean (Gao et al, 2007;Wu et al, 2008;Yang et al, 2009a;2009b;Liu et al, 2009). This simple direct transformation method does not require the Agrobacterium Ti plasmid or vector, and does not result in any unnecessary backbone or selective marker in the transgenic products.…”

Section: Introductionmentioning

confidence: 99%

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Optimization of soybean (Glycine max (L.) Merrill) in planta ovary transformation using a linear minimal gus gene cassette

Liu

Yang

Cheng

et al. 2009

J. Zhejiang Univ. Sci. B

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Abstract:Soybean transformation by ovary-drip was improved by optimizing the length of the transformation pathway by cutting the styles. These modifications facilitated soybean transformation manipulation and improved transformation reproducibility and efficiency. Using a linear minimal gus gene cassette as the foreign DNA, a maximum transformation frequency of 11% was obtained in flowers of the soybean cultivar 'Liaodou 14' with their styles mostly removed, whereas removal of only the stigma, partial style cutting and partial ovary cutting gave transformation frequencies of 0%, 1%, and 2%, respectively. An average transformation frequency of 8.2% was obtained when 619 flowers from three soybean cultivars ('Liaodou 14', 'Liaodou 13', and 'Tiefeng 29') were transformed by this optimized method. Southern blotting analysis showed that the gus reporter gene (encoding β-glucuronidase) was stably inherited with a simple pattern. Reverse transcription-polymerase chain reaction (RT-PCR) and GUS staining confirmed the expression of the gus gene in transgenic plants.

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Section: Preparation Of Dna Cassettes For Ovary Transformationmentioning

confidence: 99%

Section: Effects Of Different Style Cutting Treatments On Soybean Ovamentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Optimization of soybean (Glycine max (L.) Merrill) in planta ovary transformation using a linear minimal gus gene cassette

Liu

Yang

Cheng

et al. 2009

J. Zhejiang Univ. Sci. B

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“…Whereas Brassica plants that flower and set seed in late spring may experience a short period of high temperature, the resulting expression of YHB might even enhance photosynthesis and delay senescence potentially leading to an overall benefit to crop yield. Successful in planta transformations were also reported from other eudicot and monocot plants, including Medicago truncatula (Trieu et al, 2000), soybean (Glycine max; Gao et al, 2007), tomato (Lycopersicon esculentum; Yasmeen et al, 2009), wheat (Zale et al, 2009), maize (Zea mays; Mu et al, 2012;Moiseeva et al, 2014), and Setaria viridis (Martins et al, 2015).…”

Section: Discussionmentioning

confidence: 84%

A Tightly Regulated Genetic Selection System with Signaling-Active Alleles of Phytochrome B

Lagarias

2016

Plant Physiol.

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Selectable markers derived from plant genes circumvent the potential risk of antibiotic/herbicide-resistance gene transfer into neighboring plant species, endophytic bacteria, and mycorrhizal fungi. Toward this goal, we have engineered and validated signaling-active alleles of phytochrome B (eYHB) as plant-derived selection marker genes in the model plant Arabidopsis (Arabidopsis thaliana). By probing the relationship of construct size and induction conditions to optimal phenotypic selection, we show that eYHB-based alleles are robust substitutes for antibiotic/herbicide-dependent marker genes as well as surprisingly sensitive reporters of off-target transgene expression.

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“…The pollentube pathway method (Zhou et al 1983) bypasses the regeneration process and is a relatively simple procedure for transformation. This method is theoretically practical for all sexual reproduction plants and has proven successful in crops such as cotton (Zhou et al 1988;Zuo et al 2002), rice (Luo and Wu 1988;Zhao et al 2003), wheat (Zeng et al 1994;Hou et al 2003), maize (Zhang et al 2005;Wu et al 2008), and soybean (Hu and Wang 1999;Gao et al 2007). …”

Section: Introductionmentioning

confidence: 99%

Ovary-drip transformation: a simple method for directly generating vector- and marker-free transgenic maize (Zea mays L.) with a linear GFP cassette transformation

Yang

Liu

2008

Planta

Self Cite

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The presence of selectable marker genes and vector backbone sequences has affected the safe assessment of transgenic plants. In this study, the ovary-drip method for directly generating vector- and selectable marker-free transgenic plants was described, by which maize was transformed with a linear GFP cassette (Ubi-GFP-nos). The key features of this method center on the complete removal of the styles and the subsequent application of a DNA solution directly to the ovaries. The movement of the exogenous DNA was monitored using fluorescein isothiocyanate-labeled DNA, which showed that the time taken by the exogenous DNA to enter the ovaries was shortened compared to that of the pollen-tube pathway. This led to an improved transformation frequency of 3.38% compared to 0.86% for the pollen-tube pathway as determined by PCR analysis. The use of 0.05% surfactant Silwet L-77 + 5% sucrose as a transformation solution further increased the transformation frequency to 6.47%. Southern blot analysis showed that the transgenic plants had low transgene copy number and simple integration pattern. Green fluorescence was observed in roots and immature embryos of transgenic plants by fluorescence microscopy. Progeny analysis showed that GFP insertions were inherited in T(1) generation. The ovary-drip method would become a favorable choice for directly generating vector- and marker-free transgenic maize expressing functional genes of agronomic interest.

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Phytase expression in transgenic soybeans: stable transformation with a vector-less construct

Cited by 33 publications

References 16 publications

Optimization of soybean (Glycine max (L.) Merrill) in planta ovary transformation using a linear minimal gus gene cassette

Optimization of soybean (Glycine max (L.) Merrill) in planta ovary transformation using a linear minimal gus gene cassette

A Tightly Regulated Genetic Selection System with Signaling-Active Alleles of Phytochrome B

Ovary-drip transformation: a simple method for directly generating vector- and marker-free transgenic maize (Zea mays L.) with a linear GFP cassette transformation

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