Developing Methods for Genome Editing in Grape and Apple
Research project 29
Project researchers: Dr. Etti Or, Dr. Leor Eshed-Williams and Dr. Moshe Flaishman

Genetic editing in fruit trees is of unique value since it allows specific modification of valuable traits while keeping cultivar without modifying cultivar identity. However, the use of stable transformation for such genetic editing is problematic, in light of the long generation time and the high heterozygosity, which does not allow the preservation of cultivar identity and the removal of a transgene by crossing. Hence, genomic editing using a transient expression of CRISPR tools appears to be the preferred approach to meet the needs of genetic editing in fruit trees.

Toward achieving such an ambitious target, our initial goals are to establish reliable protocols for efficient transformation, regeneration and genome editing via CRISPR/Cas9 technology, using table grape and apple as models. Once the genetic transformation, plant transformation and CRISPR/Cas – mediated editing in grapevine and apple are well established, different approaches for transient CRISPR/Cas expression will be examined. In grapevine, we will use pre-embryonic culture to allow editing from a single cell. Because the transient transformation of protoplast by Cas9 protein is an attractive alternative for single-cell editing, we will try to establish protocols for grapevine protoplast transformation, transient editing and plantlet vine regeneration.

In grapevine, our objective is to generate dominant mutation in the VvDELLA1 gene via CRISPR-directed gene editing to produce the compact grapevine with alternative growth and yield patterns. In apple, we will attempt to disrupt dormancy-associated MdDAM genes to generate low chill apple cultivars.

Grapevine (Vitis Vinifera, cv. Prime) pre-embryogenic cells following 35Sp:GFP agro-transformation.
Grape protoplasts isolated from pre-embryogenic cells.
Apple shoots propagated for apple transformation, cv. Galaxy.
Grape pre-embryogenic culture regeneration, cv. Prime.

Dr. Etti Or 
Project researcher

Plant Sciences, Fruit Tree Sciences,
Agricultural Research Organization, Volcani Institute.

אתי אור

Research areas:

  • Grapevine molecular physiology. Grapevine bud development, mechanism of grapevine bud dormancy release and regulation of grapevine fruitfulness.
  • Regulation of berry size.

Dr. Leor Eshed-Williams 
Project researcher

Faculty of Agriculture, Food and Environment,
The Hebrew University of Jerusalem.

Research areas:

  • Meristem development and the mechanism of regeneration, mainly from the chromatin perspective.

Dr. Moshe Flaishman 
Project researcher

Plant Sciences, Fruit Tree Sciences,
Agricultural Research Organization, Volcani Institute.

Research areas:

  • Physiological and molecular approaches toward understanding flower and fruit development in deciduous fruit trees.
  • Development of biotechnological improvements of Apple, pear and fig fruits.

Research partners:

Dr. Elena Marchevka

Dr. Elena Marchevka
Ph.D. – Plant genome editing

Email: elenam@volcani.agri.gov.il

Research areas:
Plant genome editing.

Institution:
Plant Sciences, Fruit Tree Sciences, Agricultural Research Organization, Volcani Institute.

לילך פרקש-חיים

Lilach Farkash-Haim
M.Sc. – Plant tissue culture specialist

Research areas:
Plant tissue culture.

Institution:
Plant Sciences, Fruit Tree Sciences,
Agricultural Research Organization, Volcani Institute.

Tatyana Okun

Tatyana Okun
Plant tissue culture specialist

Research areas:
Plant tissue culture.

Institution:
Plant Sciences, Fruit Tree Sciences,
Agricultural Research Organization, Volcani Institute.

מיי קלסובסקי

May Klasovsky
M.Sc. student

Research areas:
Protoplast production, transformation and regeneration.

Institution:
The Hebrew University of Jerusalem.