Microprojectile-mediated genetic transformation and regeneration of Chinese elm

2003 ◽  
Vol 83 (3) ◽  
pp. 587-591 ◽  
Author(s):  
A. N. Aziz ◽  
R. J. Sauvé ◽  
S. Zhou ◽  
X. Meng
Keyword(s):  
Genetic Transformation ◽  
Cell Suspension ◽  
Cell Suspension Cultures ◽  
Polymerase Chain Reaction Analysis ◽  
Suspension Cultures ◽  
Naphthaleneacetic Acid ◽  
Glucuronidase Activity ◽  
Uida Reporter Gene ◽  
Semi Solid ◽  
Chinese Elm

A protocol for the genetic transformation and regeneration of Chinese elm ( Ulmus parvifolia Ja cq.) is reported. Cell suspension cultures were initiated from hypocotyls and were maintained on Murashige and Skoog (MS) basal semi-solid medium supplemented with varying concentrations of thiadiazuron (TDZ) and 1-naphthaleneacetic acid (NAA). Two week- old cell suspension cultures were bombarded with gold particles coated with plasmid harboring the Basta®resistance and â -glucuronidase genes. Elm cultures produced green calli within a month on selection media containing 20 mg L-1phosphinothrycin. These c alli were transferred to the same medium without the herbicide and were regenerated into shoots within 2 mo. The expression of uidA reporter gene in transformed elms was detected by histochemical analysis for â-glucuronidase activity. The presence of the Basta®resistance insert was demonstrated by polymerase chain reaction analysis. Key words:

scholarly journals Agrobacterium-Mediated Genetic Transformation and Cloning of Reference Genes in Suspension Cells of Artemisia Pallens

2021 ◽  
Author(s):  
Phanikanth Jogam ◽  
Dulam Sandhya ◽  
Anshu Alok ◽  
Mahipal Singh Shekhawat ◽  
Venkataiah Peddaboina ◽  
...  
Keyword(s):  
Heat Shock ◽  
Genetic Transformation ◽  
Cell Suspension ◽  
Reference Genes ◽  
Transformation Efficiency ◽  
Cell Suspension Cultures ◽  
Suspension Cultures ◽  
Plant Genome ◽  
Suspension Cells ◽  
Artemisia Pallens

Abstract A reliable and stable Agrobacterium-mediated genetic transformation system has been developed using cell suspension cultures derived from Artemisia pallens cotyledon explants. Cotyledon, attached cotyledon, and compound leaf were found to be suitable for the induction of callus among five different types of explants tested. Yellow friable callus derived from attached cotyledon was used to initiate suspension cultures in Suspension Culture Medium (SCM) which was supplemented with 2.4-dichlorophenoxyacetic acid (2,4-D) and in combination with different concentrations of Zeatin (ZEA). Among the two different shock treatments, cold shock (at 4 oC) for 20 minutes and heat shock (at 45oC) treatment for 5 minutes, heat shock treatment increased the transformation efficiency. Supplementation of chemical additives such as Silwet L-77 (0.05%) and Pluronic F-68 (0.05%) significantly enhanced suspension cultures' transformation efficiency. The maximum GUS intensity was recorded with an optimal intensity of blue spots in the transformed cells. The highest GUS fluorometric activity was measured as 879.4±113.7 nmol 4MU/mg/min in transformed cell suspension cultures. The hygromycin-resistant callus derived from micro-calli showed intense blue colour in GUS histochemical assay. The transgene integration into the plant genome was confirmed by polymerase chain reaction (PCR) using uidA specific primers in six hygromycin-resistant cell lines. The cloned and mRNA expression levels of three candidate reference genes ADP-ribosylation factor (Arf), β-actin (Act), and ubiquitin (Ubi), and carotenoid biosynthesis pathway gene, i.e., Phytoene desaturase (Pds) along with transgene (uidA) were evaluated in transgenic callus lines. The present Agrobacterium-mediated genetic transformation protocol could help in better understand the metabolic pathways of this medicinally important plant and its genetic improvement.

Quantitative analysis of hinokiol from cell suspension cultures of Cephalotaxus fortunei

2011 ◽  
Vol 29 (6) ◽  
pp. 567-570
Author(s):  
Xiaohui XU ◽  
Wei ZHANG ◽  
Changhong YAO ◽  
Xupeng CAO ◽  
Song XUE
Keyword(s):  
Quantitative Analysis ◽  
Cell Suspension ◽  
Cell Suspension Cultures ◽  
Suspension Cultures
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