Development and validation of protocols for product stewardship in transgenic white clover (Trifolium repens L.): detection of the AMV CP and npt2 transgenes in seeds, herbage and hay

2015 ◽  
Vol 66 (10) ◽  
pp. 1039 ◽  
Author(s):  
S. Panter ◽  
A. Mouradov ◽  
K. F. Smith ◽  
G. Spangenberg
Keyword(s):  
Mosaic Virus ◽  
White Clover ◽  
Trifolium Repens ◽  
Alfalfa Mosaic Virus ◽  
Natural Forms ◽  
Viral Coat ◽  
Ryegrass Hay ◽  
Trifolium Repens L ◽  
Coat Protein Mediated Resistance ◽  
Development And Validation

White clover (Trifolium repens L.) is an important pasture legume in temperate areas throughout the world, providing fodder for grazing animals and improving soil fertility via symbiotic nitrogen fixation. However, the persistence and stress tolerance of white clover is affected by several viruses, chiefly Alfalfa mosaic virus (AMV), Clover yellow vein virus (ClYVV) and White clover mosaic virus (WClMV). Efforts to introgress natural forms of virus resistance from other Trifolium spp. into white clover and lucerne (alfalfa) have had only limited success. This has been addressed by developing white clover germplasm exhibiting viral-coat-protein-mediated resistance to AMV and non-transgenic resistance to ClYVV. This report describes PCR-based assays for detecting the transgenes associated with the H6 transformation event in seeds, fresh leaves, air-dried leaves and mixtures of air-dried herbage of white clover and perennial ryegrass (hay). Although further development is required to convert these assays for use in the field, this paper demonstrates the ability to detect these transgenes in a range of agricultural products associated with the commercial use of white clover.

The effect of white clover mosaic virus on nodulation of white clover (Trifolium repens L. cv. Ladino)

1980 ◽  
Vol 31 (2) ◽  
pp. 307 ◽  
Author(s):  
P Guy ◽  
A Gibbs ◽  
K Harrower
Keyword(s):  
Virus Infection ◽  
Mosaic Virus ◽  
White Clover ◽  
Trifolium Repens ◽  
Effective Strain ◽  
Plant Weight ◽  
Trifolium Repens L ◽  
White Clover Mosaic Virus ◽  
Infected Nodule ◽  
Mosaic Virus Infection

White clover mosaic virus infection of Ladino clover seedlings resulted in decreased plant weight and a 71 % decrease in the number of nodules per plant without the decrease in nodule size which is usually observed when legumes are virus-infected. Nodule numbers decreased both on plants nodulated with an effective strain and on those with an ineffective strain of Rhizobium.

Development and validation of protocols for product stewardship in transgenic white clover (Trifolium repens L.): detection of the AMV CP and npt2 transgenes in pollen, honey and honey bees

2015 ◽  
Vol 66 (5) ◽  
pp. 474 ◽  
Author(s):  
S. Panter ◽  
A. Mouradov ◽  
K. F. Smith ◽  
G. Spangenberg
Keyword(s):  
White Clover ◽  
Honey Bees ◽  
Genetically Modified ◽  
Marker Gene ◽  
Alfalfa Mosaic Virus ◽  
Economic Benefits ◽  
Livestock Farmers ◽  
Trifolium Repens L ◽  
Virus Coat ◽  
Development And Validation

There are no current commercial releases of genetically modified white clover, but several research groups are working on traits such as virus resistance, stress tolerance and bloat safety that are likely to provide large economic benefits for livestock farmers. However, white clover pollen is a common constituent of honey produced by bees foraging white clover flowers. Therefore, there is a need to develop tools to detect the presence of genetically modified pollen in white clover honey. The results presented in this paper describe the development and application of PCR-based techniques to detect the Alfalfa mosaic virus coat protein gene (AMV CP) and the neomycin phosphotransferase 2 selectable marker gene (npt2) in genetically modified white clover pollen, whether this pollen is collected fresh, from honey bees that have been foraging white clover, or from honey. Further research and development will be required to develop ‘field-ready’ tools for the detection and quantification of these transgenes in pollen and honey products. However, this paper demonstrates prospects and principles in pollen and honey from honeybees foraging transgenic white clover.

scholarly journals First Report of Alfalfa mosaic virus Associated with Severe Mosaic and Mottling of Pepper (Capsicum annuum) and White Clover (Trifolium repens) in Oklahoma

Plant Disease ◽  
2012 ◽  
Vol 96 (11) ◽  
pp. 1705-1705 ◽  
Author(s):  
O. A. Abdalla ◽  
A. Ali
Keyword(s):  
United States ◽  
Mosaic Virus ◽  
Capsicum Annuum ◽  
White Clover ◽  
Trifolium Repens ◽  
Alfalfa Mosaic Virus ◽  
The United States ◽  
Morphological Data ◽  
Total Rna ◽  
First Report

Alfalfa mosaic virus (AMV), a member of the genus Alfamovirus, family Bromoviridae (1), has been reported in 44 states in the United States excluding Oklahoma. During a cucurbit survey in the summer of 2010, severe mosaic and mottling symptoms were observed on many peppers (Capsicum annuum) and white clover (Trifolium repens) plants in Tulsa, Oklahoma. Symptomatic leaf samples from 15 pepper and two white clover plants were collected in the Bixby area and analyzed serologically by dot-immunobinding assay (DIBA) using specific polyclonal antibodies against AMV (Agdia, Inc). Seven out of 15 pepper samples and both white clover samples were tested positive by DIBA to AMV. The remaining symptomatic samples were positive to Cucumber mosaic virus (CMV). Total RNA was extracted from DIBA positive AMV samples by Tri-reagent method. A small aliquot of total RNA was tested by reverse transcription (RT)-PCR using specific primers: AMV-F 5′ GTCCGCGATCTCTTAAAT 3′ and AMV-R 5′ GAAGTTTGGGTCGAGAGA 3′ that were designed to amplify 900 bp of the AMV-RNA 3. Analysis of the PCR products on agarose gel electrophoreses showed that all tested samples showed a band of the expected size while DIBA negative AMV samples did not produce any band. The amplified PCR product (900 bp) obtained from pepper and white clover were cleaned with PCR purification kit (Qiagen, Germantown, MD) and directly sequenced bi-directionally using the above primers. Sequence analysis confirmed that this virus shared 97% identity at nucleotide sequence with RNA 3 of AMV isolate from Madison-USA (GenBank Accession No. K02703). For biological and morphological characterization of the virus, eight pepper plants were mechanically inoculated using 0.1 M K2HPO4 buffer (pH 7.2) with total RNA extracted from AMV positive pepper or white clover plant samples. One to two weeks post-inoculation, all inoculated plants produced severe mosaic, mottling, and stunting. Virus-like particles preparations were obtained from these symptomatic plants according to our previously described method (2) and electron microcopy examination showed typical AMV particles. These biological and morphological data further confirmed the presence of AMV infecting pepper and clover in Oklahoma. AMV is a significant pathogen worldwide and infects more than 600 species in 70 families, especially alfalfa, pepper, soybean, and tobacco (3). AMV has a worldwide distribution, including the United States, and particularly the Midwestern U.S. where the incidence of the virus is on the rise recently because of the presence of its vector (Aphis glycines) (4). To our knowledge, this is the first report of AMV infecting crops in Oklahoma, which could pose a threat to other economic crops grown in Oklahoma, especially soybean. References: (1) E. E. Mueller et al. Plant Dis. 91:266, 2007. (2) A. Ali et al. Plant Dis. 96:243, 2012. (3) J. F. Bol. Mol. Plant Path.4:1, 2003. (4) M. Malapi-Nelson et al. Plant Dis.93:1259, 2009.

An SSR and AFLP molecular marker-based genetic map of white clover (Trifolium repens L.)

Plant Science ◽  
2003 ◽  
Vol 165 (3) ◽  
pp. 531-539 ◽  
Author(s):  
Elizabeth S. Jones ◽  
Leonie J. Hughes ◽  
Michelle C. Drayton ◽  
Michael T. Abberton ◽  
Terry P.T. Michaelson-Yeates ◽  
...  
Keyword(s):  
Molecular Marker ◽  
Genetic Map ◽  
White Clover ◽  
Trifolium Repens ◽  
Trifolium Repens L

Validation of in silico-predicted genic SNPs in white clover (Trifolium repens L.), an outbreeding allopolyploid species

2007 ◽  
Vol 277 (4) ◽  
pp. 413-425 ◽  
Author(s):  
N. O. I. Cogan ◽  
M. C. Drayton ◽  
R. C. Ponting ◽  
A. C. Vecchies ◽  
N. R. Bannan ◽  
...  
Keyword(s):  
White Clover ◽  
Trifolium Repens ◽  
In Silico ◽  
Trifolium Repens L

Observations on resistance in white clover (Trifolium repens L.) to the stem nematode (Ditylenchus dipsaci (Kühn) Filipjev)

1988 ◽  
Vol 110 (1) ◽  
pp. 145-154 ◽  
Author(s):  
R. Cook ◽  
D. R. Evans
Keyword(s):  
White Clover ◽  
Trifolium Repens ◽  
Significant Positive Correlation ◽  
Leaf Size ◽  
Nematode Reproduction ◽  
Long Period ◽  
Plant Collections ◽  
Ditylenchus Dipsaci ◽  
Trifolium Repens L ◽  
Selection For

SummaryThe expression of symptoms of stem nematode reproduction on a total of 53 white clovers (26 cultivars, 14 genepools and 13 introductions from plant collections) was studied in a series of field and glasshouse experiments. Seedlings or stolon-tip cuttings were inoculated with nematodes and the clovers classified by the proportion of plants which developed symptoms. Significant differences were found between varieties although in each test the majority was intermediate between more resistant and susceptible extremes. There was significant positive correlation between tests, in spite of different inoculation methods and different average levels of susceptibility. Very large-leaved cv. Aran was more resistant than most other clovers tested, and small-leaved cv. S. 184 was more susceptible. There was no general correlation of leaf size with reaction to stemnematode. Small-leaved cv. Pronitro was also resistant while several large-leaved cultivars were susceptible. In observations of plants exposed to nematodes over a long period, either by sequential inoculations or through perpetuating latent infections, apparently resistant plants eventually succumbed and supported nematode reproduction. It has not been determined whether this was because selection for virulence in the nematodes had occurred.

Sign in / Sign up

Export Citation Format

Share Document