Agrobacterium -mediated genetic transformation of California poppy, Eschscholzia californica Cham., via somatic embryogenesis

2000 ◽  
Vol 19 (10) ◽  
pp. 1006-1012 ◽  
Author(s):  
S.-U. Park ◽  
P. J. Facchini
Keyword(s):  
Somatic Embryogenesis ◽  
Genetic Transformation ◽  
Eschscholzia Californica ◽  
California Poppy

scholarly journals Towards A Genetic Model Organism: An Efficient Method For Stable Genetic Transformation of Eschscholzia Californica (Ranunculales)

2021 ◽  
Author(s):  
Dominik Lotz ◽  
Jafargholi Imani ◽  
Katrin Ehlers ◽  
Annette Becker
Keyword(s):  
Genetic Transformation ◽  
Genetic Model ◽  
Function Analysis ◽  
Model Organism ◽  
Protoplast Isolation ◽  
Model Systems ◽  
Eschscholzia Californica ◽  
Genetic Manipulations ◽  
California Poppy ◽  
Gene Function Analysis

Abstract California poppy (Eschscholzia californica) is a member of the Ranunculales, the sister order to all other eudicots and as such in a phylogenetically highly informative position. Ranunculales are known for their diverse floral morphologies and biosynthesis of many pharmaceutically relevant alkaloids. E. californica it is widely used as model system to study the conservation of flower developmental control genes. However, within the Ranunculales, options for stable genetic manipulations are rare and genetic model systems are thus difficult to establish. Here, we present a method for the efficient and stable genetic transformation via Agrobacterium tumefaciens-mediated transformation, somatic embryo induction, and regeneration of E. californica. Further, we provide a rapid method for protoplast isolation and transformation. This allows the study of gene functions in a single-cell and full plant context to enable gene function analysis and modification of alkaloid biosynthesis pathways by e.g. genome editing techniques providing important genetic resources for the genetic model organism E. californica.

scholarly journals Improvement of a Genetic Transformation System and Preliminary Study on the Function of LpABCB21 and LpPILS7 Based on Somatic Embryogenesis in Lilium pumilum DC. Fisch

2020 ◽  
Vol 21 (18) ◽  
pp. 6784
Author(s):  
Shengli Song ◽  
Rui Yan ◽  
Chunxia Wang ◽  
Jinxia Wang ◽  
Hongmei Sun
Keyword(s):  
Somatic Embryogenesis ◽  
Genetic Transformation ◽  
Somatic Embryo ◽  
Asymmetric Distribution ◽  
Embryo Induction ◽  
Transgenic Lines ◽  
Preliminary Study ◽  
Genetic Transformation System ◽  
Mutant Lines ◽  
Somatic Embryo Induction

Auxin transport mediates the asymmetric distribution of auxin that determines the fate of cell development. Agrobacterium-mediated genetic transformation is an important technical means to study gene function. Our previous study showed that the expression levels of LpABCB21 and LpPILS7 are significantly up-regulated in the somatic embryogenesis (SE) of Lilium pumilum DC. Fisch. (L. pumilum), but the functions of both genes remain unclear. Here, the genetic transformation technology previously developed by our team based on the L.pumilum system was improved, and the genetic transformation efficiency increased by 5.7–13.0%. Use of overexpression and CRISPR/Cas9 technology produced three overexpression and seven mutant lines of LpABCB21, and seven overexpression and six mutant lines of LpPILS7. Analysis of the differences in somatic embryo induction of transgenic lines confirmed that LpABCB21 regulates the early formation of the somatic embryo; however, excessive expression level of LpABCB21 inhibits somatic embryo induction efficiency. LpPILS7 mainly regulates somatic embryo induction efficiency. This study provides a more efficient method of genetic transformation of L. pumilum. LpABCB21 and LpPILS7 are confirmed to have important regulatory roles in L. pumilum SE thus laying the foundation for subsequent studies of the molecular mechanism of Lilium SE.

scholarly journals Induction of somatic embryogenesis in immature seeds of guavatree cv. Paluma

2009 ◽  
Vol 31 (2) ◽  
pp. 507-511 ◽  
Author(s):  
Elisa Ferreira Moura ◽  
Sérgio Yoshimitsu Motoike
Keyword(s):  
Ascorbic Acid ◽  
Somatic Embryogenesis ◽  
Genetic Transformation ◽  
Embryogenic Callus ◽  
Cellular Proliferation ◽  
Psidium Guajava ◽  
Embryogenic Potential ◽  
Immature Seeds ◽  
Different Types ◽  
Secondary Embryos

The biotechnological techniques may help solve many problems of guava culture, such as the high perishability of fruits. Somatic embryogenesis can generate highly multiplicative cell cultures and with high regenerative potential, serving as basis for genetic transformation. The aim of this work was to obtain somatic embryogenesis of guava (Psidium guajava L.) cv. Paluma. Immature seeds were used, and they were inoculated in MS environment containing 400 mg L-1 of L-glutamine, 100 mg L-1 myo-inositol, 60 g L-1 sucrose, 100 mg L-1 ascorbic acid and supplemented with different types and concentrations of growth regulators. Embryogenic callus appeared after 37 days of culture in an environment containing 1.0 mg L-1 2.4-D + 2.0 mg L-1 2-ip, in 7% of the explants. After 65 days of culture, the treatment containing 0.5 mg L-1 CPA showed 20% of explants with direct embryos, while the treatment with 1 mg L-1 had 14% of explants with direct embryos and 7% of explants with embryogenic callus. In 66.6% of embryos regenerated with 0.5 mg L¹ CPA there was the formation of secondary embryos. The use of IASP and BAP, aiming embryogenesis proliferation, led to an increase in the cellular proliferation, but calli apparently lost their embryogenic potential.

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