Pathogen-resistant grape plants

a technology resistant grapes, which is applied in the field of pathogen-resistant grape plants, can solve the problems of limited genetic improvement of grapes through conventional breeding, high heterozygous cultivated gaps, and inability to generally breed true, etc., to achieve enhanced embryogenic culture initiation frequency and facilitate an extraordinarily high frequency of somatic embryo formation

Inactive Publication Date: 2006-02-16
SUBRAMANIAN JAYASANKAR +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0005] We have discovered methods for growing perennial grape embryogenic cultures and for growing large quantities of somatic grape embryos from such perennial embryogenic cultures in a relatively short period using a liquid suspension culture. Several advantages are provided by the present methods. These approaches, for example, facilitate an extraordinarily high frequency of somatic embryo formation and plant regeneration. Such frequencies have not been previously reported for grapevine regeneration of any known cultivar, and render the method useful for large-scale production of clonal planting stock of grape plants. In addition, the methods produce embryos free of such common abnormalities as fusion and fasciations of somatic embryos. The methods of the invention also result in enhanced embryogenic culture initiation frequency, allowing for the production of highly embryogenic cultures that can then be successfully carried through the subsequent stages of the regeneration process to the whole plant level. Because of these advantages, the methods of the invention are especially useful in the application of biotechnology for the genetic improvement of this crop.
[0022] In a fifteenth aspect, the invention features a method for decreasing pathogen-mediated damage to a plant, the method including contacting the plant with a recombinant form of a protein that exhibits increased level of expression following contact with a pathogen.
[0056] By “PR-5” is meant a protein that is substantially identical to VVTL-1 (SP accession no. 004708) and, when overexpressed in a grape plant, confers on the plant increased pathogen resistance.

Problems solved by technology

Current methods for improving grapevines are time-consuming and labor intensive.
For example, genetic improvement in grapes through conventional breeding is severely limited by a number of factors such as long pre-bearing age and varying ploidy levels.
Cultivated gapes are also highly heterozygous and do not generally breed true from seeds.
Moreover, grape breeding programs are expensive, long-term products.

Method used

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Examples

Experimental program
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Effect test

example 1

Perennial Grape Embryonic Culture System

[0088] The following method has proven effective for the production of perennial embryogenic grape cultures, and for the regeneration of grapevine by somatic embryogenesis.

[0089] Explant Tissue and Culture Initiation

[0090] In the culture initiation step, explant material was collected from the field, greenhouse, or in vitro shoot micropropagation cultures of grapevine and placed into in vitro culture. This explant material was typically collected from leaves, anthers, or tendrils, but is also obtained from other vegetative or reproductive tissues of grapevine. Once collected, the explant tissue, if desired, was surfaced sterilized according to standard methods, and then placed on a suitable solid culture initiation medium in a petri plate.

[0091] Any of a number of well known media, e.g., Murashige and Skoog (MS) and Nitsch's medium, may be used. Such media typically include inorganic salts, vitamins, micronutrients, a nitrogen source, and ...

example 2

Production of Highly Embryogenic Grape Cells Using Liquid Suspension or Solid Cultures

[0102] A method has also been developed for the production of large quantities of grapevine somatic embryos using either a liquid cell suspension culture or a solid culture system. These methods are particulary useful for producing highly embryogenic cells that are capable of regenerating into whole plants. Below, a simple protocol for efficient somatic embryogenesis of grapevine using either a liquid cell suspension culture or a solid culture system is presented.

[0103] In general, the method includes a multistage culturing process typically involving (i) culture initiation; (ii) identification and isolation of embryogenic cells or embryogenic cell masses; (iii) production of perennial embryogenic cultures; and (iv) concentration of highly embryogenic cell clusters. The method involves the following steps.

[0104] Explant tissue is placed on a suitable culture initiation medium, as is described he...

example 3

Selection of Disease Resistant Embryogenic Cells and Plants of Grapevine

[0141] The perennial grape embryogenic cultures of the invention can be used for the selection or screening for grape cells having resistance to toxic substances, such as those present in a filtrate produced by a fungal culture. Such pathogens include, without limitation, bacteria and fungi. Plant diseases generally caused by these pathogens are described in Chapters 11-16 of Agrios, Plant Pathology, 3rd ed., Academic Press, Inc., New York, 1988, hereby incorporated by reference. The “Compendium of Grape Diseases” (APS Press (1988) R. C. Pearson & A. C. Goheen, Eds.) describes diseases that affect grape plants. Examples of bacterial pathogens include, without limitation, Agrobacterium vitis, Agrobacterium tumefaciens, Xylella fastidosa, and Xanthomonas ampelina. Examples of fungal pathogens include, without limitation, Plasmopara viticola, Botrytis cinerea, Guignardia bidwellii, Phomophsis viticola, Elsinoe amp...

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Abstract

The invention features a method of producing a grape somatic embryo having resistance to a plant pathogen, the method including the steps of (a) culturing a grape somatic embryo in a first liquid culture medium that includes a plant growth regulator and a phytotoxin from a plant pathogen; (b) exchanging the first liquid culture medium for a second liquid culture medium not including the phytotoxin; (c) recovering a living grape cell or grape cell cluster from the second liquid culture, the living cell or cell cluster being resistant to the pathogen; and (d) culturing the grape cell or grape cell cluster in a third culture medium to produce a grape somatic embryo.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application is a continuation of U.S. Ser. No. 09 / 570,217 filed May 12, 2000, which claims the benefit to U.S. Provisional Application Nos. 60 / 134,275, filed May 14, 1999 and 60 / 148,251, filed Aug. 11, 1999, each of which is hereby incorporated by reference.BACKGROUND OF THE INVENTION [0002] This invention relates to plants having an increased level of resistance to a pathogen and to methods for producing them. [0003] Grapevines are a deciduous temperate fruit crop of ancient origin. Grape production (65×106 metric tons) exceeds that of any other temperate fruit crop, and ranks third after Citrus and banana production. In addition, due to its uses for fresh fruit, juice, jelly, raisins, and wine, grapes surpass all other fruit crops in value. Therefore, successful efforts to improve grapevines are likely to have a major impact on commercial viticulture. [0004] Current methods for improving grapevines are time-consuming and labor in...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12N15/82C12N5/00C12N15/87A01H1/00A01HA01H5/00C07KC07K14/415C12NC12N5/10C12N15/29
CPCC12N15/8282C07K14/415
Inventor SUBRAMANIAN, JAYASANKARLI, ZHIJIANGRAY, DENNIS
Owner SUBRAMANIAN JAYASANKAR
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