Method for cultivation of genetically-modified plant

Patent metadata

Field	Value
Publication number	US-9353379-B2
Application number	US-200913147436-A
Country	US
Kind code	B2
Filing date	Aug 26, 2009
Priority date	Feb 2, 2009
Publication date	May 31, 2016
Grant date	May 31, 2016

Abstract

The present invention provides a method for cultivation of a genetically-modified plant that can highly produce a desired protein. More specifically, the present invention provides a method for cultivation of a genetically-modified plant comprising: cultivating the genetically-modified plant in a medium,
wherein the genetically-modified plant is transformed by introducing an expression vector comprising a promoter regulating expression of RNA and a seed storage protein isolated from a plant that highly expresses RNA and the seed storage protein under a high nitrogen condition; and a polynucleotide encoding an objective protein, and wherein the medium is adjusted so that nitrate nitrogen is 70 mg/L to 750 mg/L and/or ammonium nitrogen is 70 mg/L to 750 mg/L for a definite period in a period from 30 days before an expected flowering date to a date on or before flowering of the genetically-modified plant.

First claim

The invention claimed is:

1. A method for hydroponic cultivation of a genetically-modified plant comprising: cultivating the genetically-modified plant in a hydroponic medium,
wherein the genetically-modified plant is transformed by introducing an expression vector comprising:
a promoter that regulates expression of RNA expressed in a seed that satisfies the formula (1):
V/W> 1.0  (1),

wherein V and W are determined as follows:
V is an amount of RNA in the seed of a predetermined plant when the predetermined plant is cultivated in a hydroponic medium adjusted so that nitrate nitrogen is 70 mg/L to 750 mg/L and/or ammonium nitrogen is 70 mg/L to 750 mg/L for a period which starts from 30 days before the expected flowering date and ends at the flowering day, and W is an amount of RNA contained in a seed of the predetermined plant when the plant is cultivated in a medium adjusted so that nitrogen is 0 mg/L to 50 mg/L for a period which starts from 30 days before the expected flowering date and ends at the flowering day, wherein the predetermined plant is the same species as the genetically-modified plant, wherein RNA is extracted from the seeds 15 to 25 days after the flowering,
wherein the promoter is a glutelin promoter; and
a polynucleotide located downstream of the promoter and encoding a protein of interest,
wherein the medium for cultivating the genetically-modified plant is adjusted so that a content of nitrate nitrogen is from 70 mg/L to 750 mg/L and a content of ammonium nitrogen is from 50 mg/L to 750 mg/L for a period which starts from 30 days before the expected flowering date and ends at the flowering day of the genetically-modified plant, and
wherein the ratio of the contents of the nitrate nitrogen to the ammonium nitrogen is from 3:1 to 1:3.

2. The method according to claim 1 , wherein the predetermined plant is a poaceous plant and the genetically-modified plant is a poaceous plant.

3. A method for production of a seed comprising cultivating the genetically-modified plant according to the method of claim 1 and collecting the seed.

4. The method according to claim 3 , wherein the plant is a rice plant and the seed is a rice seed.

5. A method for hydroponic cultivation of a genetically-modified plant comprising:
cultivating the genetically-modified plant in a hydroponic medium,
wherein the genetically-modified plant is transformed by introducing an expression vector comprising:
a promoter that regulates expression of RNA expressed in a seed, wherein the promoter is a glutelin promoter; and
a polynucleotide located downstream of the promoter and encoding a protein of interest,
wherein the medium for cultivating the genetically-modified plant is adjusted so that a content of nitrate nitrogen is from 70 mg/L to 750 mg/L and a content of ammonium nitrogen is from 50 mg/L to 750 mg/L for a period which starts from 30 days before the expected flowering date and ends at the flowering day of the genetically-modified plant, and
wherein the ratio of the contents of the nitrate nitrogen to the ammonium nitrogen is from 3:1 to 1:3.

Assignees

• Kasahara Saori
• Wasai Masafumi
• Sugita Koichi
• Shimada Teruhisa
• Jujo Paper Co Ltd

Inventors

• Kasahara Saori
• Wasai Masafumi
• Sugita Koichi
• Shimada Teruhisa

Classifications

• C12N15/8251

  Amino acid content, e.g. synthetic storage proteins, altering amino acid biosynthesis · CPC title

• C12N15/8238

  chemically inducible, e.g. tetracycline · CPC title

• C12N15/8257

  for the production of primary gene products, e.g. pharmaceutical products, interferon · CPC title

• C12N15/8261 Primary

  with agronomic (input) traits, e.g. crop yield · CPC title

• C12N15/8234

  Seed-specific, e.g. embryo, endosperm · CPC title