Modulation of sugar and amino acid content (SULTR3) in plants

A plant and plant cell technology, applied in plant genetic improvement, botanical equipment and methods, plant peptides, etc., can solve problems such as limited opportunities for tobacco products and limited types of tobacco

Pending Publication Date: 2022-05-17
PHILIP MORRIS PROD SA
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, the limited variety of tobacco available for commercial production means that there are limited opportunities to develop tobacco products with different flavors and aromas

Method used

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  • Modulation of sugar and amino acid content (SULTR3) in plants
  • Modulation of sugar and amino acid content (SULTR3) in plants
  • Modulation of sugar and amino acid content (SULTR3) in plants

Examples

Experimental program
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Effect test

preparation example Construction

[0215] 6. Preparation, screening and hybridization of modified plants

[0216] NtSULTR3 or NtSULTR3 and NtSUS polynucleotides prepared from individual plants, plant cells or plant material may optionally be combined to expedite screening for mutations in plant populations derived from mutagenized plant tissues, cells or material. One or more subsequent generations of plants, plant cells or plant material can be screened. The size of the optionally pooled cohort depends on the sensitivity of the screening method used.

[0217] After the samples are optionally pooled, they can be subjected to polynucleotide-specific amplification techniques, such as PCR. Any one or more primers or probes specific for the gene or for sequences immediately adjacent to the gene can be used to amplify the sequences within the optionally pooled samples. Suitably, one or more primers or probes are designed to amplify regions of the locus where useful mutations are most likely to occur. Most prefera...

Embodiment 1

[0320] Example 1 - Materials and methods

[0321] DNA extraction and plant genotyping

[0322] Leaf samples were extracted using BioSprint 96 (Qiagen, Hilden, Germany) and the BioSprint 96 DNA Plant Kit (Qiagen, Hilden, Germany). DNA samples were used in TaqMan reactions to determine plant genotypes. Taqman was performed using the ABI PRISM 7900HT Sequence Detection System (Applied Biosystems, Life Technologies, Foster City, CA, USA) and TaqMan Rapid Advanced Master Mix (Applied Biosystems, Foster City, CA, USA).

[0323] Measuring Free Amino Acid Content

[0324] Amino acid content can be measured using various methods known in the art. One such method is Method MP 1471 rev 5 2011, Resana, Italy: Chelab Silliker S.r.l, Mérieux NutriSciences Company. To determine the amino acids in dried plant leaves, the dried leaves were dried at 40°C for 2-3 days after removal of the midrib, if necessary. The tobacco material was then ground to a fine powder (approximately 100 uM) prio...

Embodiment 2

[0327] Analysis of the expression of embodiment 2-NtSULTR gene

[0328] Table 1 shows that NtSULTR3;3-T is expressed throughout plant tissues, especially in petals. Interestingly, copies of NtSULTR3;3-S are not expressed in N. virginica, but in some other tobaccos, such as TN90. Apparently, the NtSULTR3;3-S genomic sequence was not identified in the Virginia tobacco genome or was altered in Virginia tobacco and dark tobacco (Sierro et al. (2014) Nat Commun. May 8;5:3833, see Table 3 and 4). The NtSULTR3;3-S genome and polypeptide sequences were deduced from the TN90 sequencing library. Other SULTR3 genes expressed in petals were NtSULTR3;1A-S, NtSULTR3;1A-T and NtSULTR3;1B-S. NtSULTR3;4A-T is apparently more specific for stems. NtSULTR3;2-S is expressed in sepals and roots. Interestingly, several SULTR3 genes were not expressed or weakly expressed in green leaves, namely, NtSULTR3; 1A-S, NtSULTR3; 1A-T, NtSULTR3; 3-S, NtSULTR3; 4A-S, NtSULTR3; 4B-S, NtSULTR3; 4B-T, NtSUL...

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Abstract

Described herein is a plant cell comprising: (i) a polynucleotide comprising at least one polynucleotide associated with SEQ ID NO: 1 (NtSULTR3; 1 A-S), SEQ ID NO: 3 (NtSULTR3; 1 A-T), SEQ ID NO: 5 (NtSULTR3; 1B-S), SEQ ID NO: 7 (NtSULTR3; 1B-T), SEQ ID NO: 15 (NtSULTR3; 3-T), SEQ ID NO: 17 (NtSULTR3; 4A-S), SEQ ID NO: 19 (NtSULTR3; 4A-T) or SEQ ID NO: 23 (NtSULTR3; 4B-T), consisting or consisting essentially of a sequence having at least 60% sequence identity; (ii) a polypeptide encoded by the polynucleotide set forth in (i); (iii) a polypeptide comprising at least one polypeptide associated with SEQ ID NO: 2 (NtSULTR3; 1A-S) has at least 87% sequence identity or is at least 87% sequence identity to SEQ ID NO: 4 (NtSULTR3; 1A-T) has at least 87% sequence identity or is at least 87% sequence identity to SEQ ID NO: 6 (NtSULTR3; 1B-S) has at least 87% sequence identity or is at least 87% sequence identity with SEQ ID NO: 8 (NtSULTR3; 1B-T) has at least 88% sequence identity or is at least 88% sequence identity with SEQ ID NO: 16 (NtSULTR3; 3-T) has at least 70% sequence identity or is at least 70% sequence identity with SEQ ID NO: 18 (NtSULTR3; 4A-S) having at least 84% sequence identity to SEQ ID NO: 20 (NtSULTR3; 4A-T) having at least 79% sequence identity; or at least one of SEQ ID NO: 24 (NtSULTR3; 4B-T), consisting or consisting essentially of a sequence having at least 87% sequence identity; or (iv) a construct, vector or expression vector comprising the isolated polynucleotide set forth in (i) wherein the plant cell comprises at least one modification as compared to a control plant cell in which the expression or activity of the polynucleotide or polypeptide is not modified, the at least one modification modulates (a) the expression or activity of the polynucleotide or (b) the expression or activity of the polynucleotide or the polypeptide.

Description

[0001] Cross References to Related Applications [0002] This application claims priority from European Patent Application No. 19200856.3 filed October 1, 2019, the disclosure of which is hereby incorporated by reference. technical field [0003] The present invention relates to plant cells and the like having regulated expression or activity of a chloroplast sulfate transporter (SULTR3). Background technique [0004] To make tobacco products, different types of tobacco are mixed in various ratios to create a blend with certain flavor characteristics. Flue-cured tobacco (eg, Virginia) is the most widely grown tobacco and is characterized by a high sugar to nitrogen ratio, but has limited flavor profiles. Other tobacco types—such as air-dried (e.g., Burley, Maryland, and Galpao) or cured (e.g., Dark) tobacco types—offer alternative flavors feature. These different flavor profiles are very important in the production of blended tobacco products. [0005] Flavor profiles a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/84C07K14/415C12N15/29A01H5/00A01H6/82
CPCC12N15/8251C12N15/8218C12N15/8245C07K14/415C12N15/8246A01H6/823A24B15/24A24B3/10
Inventor L·博维特
Owner PHILIP MORRIS PROD SA
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