- Core Citrus Transformation Facility
Transgenic Citrus Order Form
IMPORTANT INFORMATION FOR PROSPECTIVE CLIENTS!
In the fight against “greening” disease (HLB) and Citrus canker transgenic plants remain one of the most promising tools. As a result, demand for transgenic Citrus plants has increased in the last 4-5 years significantly. That is also reflected in the number of orders CCTF received and still does on a regular basis. Considering the magnitude of the impact these two diseases have on Citrus industry, researchers are racing to get potentially useful products out to growers as soon as possible. That has created a shift in what type of plants are being generated by CCTF and how. For the purpose of determining if certain gene (or DNA construct) is beneficial to scion cultivar, researchers are advised to use Duncan grapefruit. This cultivar is very susceptible to both HLB and canker, it is not difficult to transform, and the fruit is seedy which allows CCTF management to have relatively stable source of starting material for co-incubation experiments. Alternative cultivar for projects done on sweet orange is Pineapple sweet orange. If potentially beneficial effect of gene on rootstock is sought, researchers are advised to use Carrizo rootstock. This cultivar is relatively easy to transform and is also very seedy. The orders coming to CCTF in this period have a goal to provide “proof of concept” data and are not destined for pipeline resulting in plants that would be commercialized. For that reason, management of CCTF suggests the use of GFP as a reporter gene. The use of GFP gene, allows timely production of transgenic plants. GUS as reporter gene remains popular with some research groups and, together with nptII gene, was de-regulated by federal government. GUS assays are more labor demanding than GFP. Decision to use GUS as a reporter gene will result in longer delivery time for the placed orders.
CCTF remains dedicated to provide best possible service to any parties involved in projects that would improve profitability of Citrus industry. However, at these testing times the priority has to be given to projects that have to do with production of tolerant/resistant citrus plants to HLB and canker. Above mentioned suggestions are derived from the experience gained by CCTF manager and are based on results of multiple experiments performed in CCTF within last 10 years. It is our hope that prospective clients will accept these suggestions for the mutual benefit of everybody involved.
1) Placing the order
It is up to clients who are placing the order to decide on how the transformation will be done. Here are the ways order can be processed:
TERMS & CONDITIONS
OPTIONS FREE OF INTELLECTUAL PROPERTY ISSUES:
- A) The "transformation ready" Agrobacterium culture is supplied. CCTF will do the transformation.
- B) The gene of interest is inserted anywhere between Right and Left border of binary vector and newly made construct(s) are sent to CCTF. We will mobilize vector(s) into Agrobacterium and do the transformation.
OPTIONS WITH INTELLECTUAL PROPERTY ISSUES:
- C) The gene of interest is sent to CCTF where insertion into binary vector will be done. We will than mobilize binary vector into Agrobacterium and do the transformation.
- D) The DNA fragment of interest is sent to CCTF where we will clone it between promoter and terminator sequences (to make it a gene-sensu stricto), and insert into binary vector. We would than mobilize binary vector into Agrobacterium and do the transformation.
Since in options C and D CCTF would have some intellectual input, ownership of intellectual property of produced plants would need to be predefined prior to initiation of the work.
Management of the CCTF has certain number of binary vectors proven to work well. We recommend utilization of those vectors for transformation experiments for any of the four options listed above.
The price for production of transgenic plants is as follows:
Each “gene” (DNA fragment of interest) per Citrus cultivar is $3,000. This will cover production of 10 plants. For each additional plant produced client will be charged additional $200. Example: if introduction of gene “A” is desired in Hamlin orange that would cost $3,000. If 14 plants are delivered than the price would be 3,000+(4x200)=$3,800. Producing both Hamlin oranges and Flame grapefruit carrying same gene “A” would cost 3,000x2=$6,000.
Clients placing the order will be expected to pay the amount for first 10 plants up-front.
This measure had to be adopted considering our previous experience. Some orders involve “gene(s)” for which effects on putative transgenic plants are not well described. In the case of genes that may have detrimental or inhibitory effects on growth, it may be very difficult and in some cases even impossible to obtain live transgenic plants. CCTF will make five attempts (five experiments with up to 5000 explants) to create desired transgenic plants. Even if we are not successful, work on such orders has to be compensated.
Special arrangements will be made for transformation with a fragment of DNA containing more the one gene.
Prices are quadrupled if insertion of gene of interest without the reporter gene is requested.
Orders coming from clients who are conducting HLB-associated research funded through NAS will be processed with the 50% discount.
To obtain the prices for clients outside of Florida, all the amounts in the table should be multiplied by two.
3) CCTF is responsible for shipping of transgenic material. However, specific permit for each individual order must be furnished prior to shipment.
4) Once this form is signed by the CCTF management, it becomes binding contract. Photocopies of signed contract will be sent back to clients for records. This order form also represents the Material Transfer Agreement. Upon receipt of biological material such as plasmid DNA and/or bacterial strains, management of CCTF becomes responsible for its use. No dissemination of such material or its use for other purposes will be attempted except if specifically permitted by the sender. Any changes to this contract must be made in writing, attached to this copy and signed by both parties.
Orders can be placed online or by mail. If you would like to order by mail, please print this order form and sign it in the box labeled client. Signed form should be sent to:
Dr. Vladimir Orbovic
Citrus Core Transformation Facility
CREC-University of Florida-IFAS
700 Experiment Station Road
Lake Alfred, 33850
When ordering transgenic plants, have in mind that success rate for the transformation varies greatly among Citrus cultivars. The best results are obtained with: Carrizo citrange rootstock cultivar and grapefruit cultivars led by Duncan, followed by Flame, Marsh, and Ruby Red. As a result of “ease” in producing transgenic plants of these cultivars, processing of these orders will take shorter amount of time. If you do not have specific cultivar requirements for research projects and want transgenic plants as soon as possible, than we would suggest picking one of the above mentioned cultivars as your choice. Out of two cultivars of sweet orange that we transform on a regular basis-Valencia and Hamlin, none of them shows large advantage over the other in transformation efficiency.
Be advised that as part of regular activities, management of CCTF presents seminars on the progress of work in the field of transgenic Citrus. Limited amount of data mostly pertaining to efficiency of transformation and factors the affect this process from almost all projects (orders) is presented. None of this information is reported in written form and if any details of particular project (order) are mentioned, names of the persons involved in leadership of the project are mentioned. If you would not like any of the information shared publicly, please indicate that clearly on the hard copy of order form that you will sign and send to CCTF.
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