PLANTS HAVING INCREASED OIL QUALITY

• US 20200131523A1
• Filed: 01/26/2018
• Published: 04/30/2020
• Est. Priority Date: 01/27/2017
• Status: Active Grant

• Current Assignee: Board of Trustees of The University of Illinois, Regents of The University of Minnesota (University of Minnesota)
• Original Assignee: Regents of The University of Minnesota (University of Minnesota)
• Inventors: MARKS, Michael David, SEDBROOK, John C., WYSE, Donald L., DORN, Kevin
• Granted Patent: US 11,396,657 B2
• Time in Patent Office: Days
• CPC Class Codes: A01H 1/06 Processes for producing mutations, e.g. treatment with chemicals or with radiation specific mutations prepared by genetic engineering on plant cell or plant tissues C12N15/00 ; process for producing transgenic plants C12N15/82
• CPC Class Codes: A01H 5/10 Seeds
• CPC Class Codes: A01H 6/20 Brassicaceae, e.g. canola, broccoli or rucola
• CPC Class Codes: C12N 15/8213 Targeted insertion of genes into the plant genome by homologous recombination
• CPC Class Codes: C12N 15/8247 involving modified lipid metabolism, e.g. seed oil composition
• CPC Class Codes: C12N 9/1029 transferring groups other than amino-acyl groups (2.3.1)
• CPC Class Codes: C12N 9/1085 transferring alkyl or aryl groups other than methyl groups (2.5)
• CPC Class Codes: C12Y 205/01 transferring alkyl or aryl groups, other than methyl groups (2.5.1)

Abstract

This document provides materials and methods for generating oilseed (e.g., pennycress) plants that having low levels of erucic acid. For example, oilseed plants having reduced expression levels of one or more polypeptides involved in erucic acid metabolism (e.g., fatty acid elongase 1 (FAE1)), as well as materials and methods for making and using oilseed plants having low levels of erucic acid are provided.

Claims

1-43. -43. (canceled)

44. A method for generating an oilseed plant having low levels of erucic acid as compared to a corresponding wild type oilseed plant, the method comprising:

introducing into an oilseed plant cell a nuclease and a guide sequence, wherein said guide sequence comprises a nucleic acid sequence specific to the fatty acid elongase 1 (FAE1) gene under conditions where said nuclease introduces a loss-of-function modification into the FAE1 coding sequence of said FAE1 gene;

selecting an oilseed plant cell having low levels of erucic acid as compared to a wild oilseed plant; and

regenerating an oilseed plant having low levels of erucic acid from said selected oilseed plant cell.

45. The method of claim 44, wherein said oilseed plant is a pennycress plant.

46. The method of claim 44, wherein said nuclease is a CRISPR associated (Cas) nuclease.

47. The method of claim 46, wherein said Cas nuclease is a Cas9 nuclease.

49. The method of claim 44, wherein said guide sequence comprises SEQ ID NO:

15.

51. The method of claim 44, wherein said loss-of-function modification in said FAE1 coding sequence comprises a 4 base-pair deletion as set forth in SEQ ID NO:

3.

54. The method of claim 44, wherein said loss-of-function modification in said modified FAE1 coding sequence encodes a truncated FAE1 polypeptide as set forth in SEQ ID NO:

4 or as set forth in SEQ ID NO;

6.

56. The method of claim 44, wherein said loss-of-function modification in said modified FAE1 coding sequence comprises a single base-pair adenine (‘

A’

) insertion as set forth in SEQ ID NO;

5.

62. The method of claim 44, wherein said low levels of erucic acid comprise less than about 2% erucic acid.

48. (canceled)

50. (canceled)

52. (canceled)

53. (canceled)

55. (canceled)

57-61. -61. (canceled)

63. An oilseed plant having low levels of erucic acid as compared to a corresponding wild type oilseed plant, said oilseed plant comprising:

a genome edited in a site-specific manner to modify the coding sequence of the fatty acid elongase 1 (FAE1) gene;

wherein said modified FAE1 coding sequence is a loss-of-function modification effective to cause low levels of erucic acid as compared to a wild type oilseed plant.

64. The oilseed plant of claim 63, wherein said oilseed plant is a pennycress plant.

65. The oilseed plant of claim 63, wherein seed oil from said low levels of erucic acid comprises less than about 5% erucic acid.

66. The oilseed plant of claim 65, wherein seed oil from said low levels of erucic acid comprises less than about 2% erucic acid.

68. The oilseed plant of claim 63, wherein said loss-of-function modification comprises a modified FAE1 coding sequence comprising a 4 base-pair deletion as set forth in SEQ ID NO:

3.

71. The oilseed plant of claim 63, wherein said modified FAE1 coding sequence encodes a truncated FAE1 polypeptide.

72. The oilseed plant of claim 71, wherein said loss-of-function modification comprises a modified FAE1 coding sequence encoding a truncated FAE1 polypeptide as set forth in SEQ ID NO:

4 or SEQ ID NO;

6.

73. The oilseed plant of claim 63, wherein said loss-of-function modification comprises a modified FAE1 coding sequence comprising a single base-pair adenine (‘

A’

) insertion as set forth in SEQ ID NO;

5.

79. The oilseed plant of claim 63, wherein seed oil from said oilseed plant comprises less than 2% eicosenoic acid.

81. The oilseed plant of claim 63, wherein seed oil from said oilseed plant comprises about 25% to about 55% oleic acid.

83. The oilseed plant of claim 63, wherein seed oil from said oilseed plant comprises about 20% to about 40% linoleic acid.

85. The oilseed plant of claim 63, wherein seed oil from said oilseed plant comprises about 13% to about 30% linolenic acid.

87. A seed produced by the oilseed plant of claim 63.

88. Seed oil produced by the oilseed plant of claim 63.

67. (canceled)

69. (canceled)

70. (canceled)

74-78. -78. (canceled)

80. (canceled)

82. (canceled)

84. (canceled)

86. (canceled)