BACKGROUND Grasses are relatively recalcitrant to genetic transformation in comparison to certain dicotyledons, yet they constitute some of the most important biofuel crops. Genetic transformation of switchgrass (Panicum virgatum L.) has previously been reported after cocultivation of explants with Agrobacterium and biolistics of embryogenic calli. Experiments to increase transient gene express...

Transient expression profiles for several chimeric beta-glucuronidase (GUS) gene constructs were determined in tissues (young leaves, mature leaves and roots) of creeping bentgrass (Agrostis palustris, cv. Penn A4) following microprojectile bombardment. The constructs analyzed consisted of the uidA (GUS) reporter gene driven by four different promoters (ubiquitin 3-potato, ubiquitin corn, ubiqu...

In order to develop an easy, rapid and efficient protocol for the genetic transformation of tomato (Lycopersicon esculentum Mill.), without involving tissue culturing, ripened fruits of tomato were infiltrated with Agrobacterium harboring plasmid pROKII GUSint AP1. The plasmid contains Arabidopsis early flowering gene AP1, GUS reporter gene and NPTII as a selectable marker gene. Both transient ...

Background: Sugarcane is a monocotyledonous crop that is cultivated in the tropical and subtropical regions of the world. One of the most important criteria, influencing the efficiency of the sugarcane transformation is known to be related to physical and biological factors during the transformation procedure. Objectives: The objective of this research was to study the response of callus induc...

A novel sequence that functions as a promoter element for moderate constitutive expression of transgenes, designated as the PtMCP promoter, was isolated from the woody perennial Populus tomentosa. The PtMCP promoter was fused to the GUS reporter gene to characterize its expression pattern in different species. In stable Arabidopsis transformants, transcripts of the GUS reporter gene could be de...

Kernel culture was assessed for evaluating novel gene expression in developing maize (Zea mays L.) seeds by comparing the transient expression of maize ubiquitin (Ubi-I) promoter-driven ,B-glucuronidase (GUS) delivered by particle bombardment in kernels grown in culture with those grown in planta. With kernels from either source, GUS expression, as determined by histochemical staining, was wide...

We have characterized the 5' region of the rice actin 1 gene (Act1) and show that it is an efficient promoter for regulating the constitutive expression of a foreign gene in transgenic rice. By constructing plasmids with 5' regions from the rice Act1 gene fused to the coding sequence of a gene encoding bacterial beta-glucuronidase, we demonstrate that a region 1.3 kilobases upstream of the Act1...

Sections of hypocotyls, roots and leaves from Pentalinon andrieuxii plantlets were transiently transformed with Agrobacterium tumefaciens LBA4404 bearing the binary plasmid pCAMBIA2301 with an interrupted β-glucuronidase (GUS) gene. Histochemical GUS assays showed transient gene expression in all infected tissues, being older roots those which displayed the most intense GUS staining. To our kno...

An airgun apparatus has been constructed for transient gene expression studies of monocots. This device utilizes compressed air from a commercial airgun to propel macroprojectile and DNA-coated tungsten particles. The beta-glucuronidase (GUS) reporter gene was used to monitor transient expression in three distinct cell types of maize (Zea mays), rice (Oryza sativa), and wheat (Triticum aestivum...

We report here an alternative and easy approach to generate sufficient quantity of minimal gene expression cassette (promoter–open reading frame–terminator) DNA by PCR amplification using high fidelity DNA polymerases. Isolating the minimal gene cassette DNA in large quantities by restriction digestion and gel extraction for biolistic-mediated ‘clean DNA’ transformation, is not only a cumbersom...