EP4366512A2

EP4366512A2 - New processed food product based on wheat grains, manufacturing methods and applications

Info

Publication number: EP4366512A2
Application number: EP22754899.7A
Authority: EP
Inventors: Pierre BERBEZY; Bruno Viallis; Bernard Duperrier; Jean-Baptiste Laffaire
Original assignee: Limagrain Ingredients SAS
Current assignee: Limagrain Ingredients SAS
Priority date: 2021-07-09
Filing date: 2022-07-08
Publication date: 2024-05-15
Also published as: WO2023281229A2; FR3125063A1; WO2023281229A3

Abstract

The invention aims to provide a new food product based on processed wheat grains that are rich in sugars. To this end, the invention relates to a food product comprising processed wheat grains, characterised in that these wheat grains have a milling yield RM, achieved in accordance with standard NF EN ISO 27971, expressed as wt.% of dry matter in relation to the whole wheat grains, of 0.5 ≤ RM ≤ 10.10. The invention also relates to a processing method for obtaining the food product according to the invention, as well as to the mutant wheat used in the invention and the applications of this food product.

Description

Novel food product processed from wheat grains, manufacturing methods and applications

Field of the invention [0001] The invention relates to the field of food, in particular human food.

[0002] In particular, the invention relates to a new processed food product based on wheat grains and its manufacturing processes.

The invention also relates to food applications from these processed wheat grains.

Technological background of the invention

[0004] Wheat designates cereals of the genus Triticum. The main outlet for wheat in the food industry is the transformation of wheat into flour (soft wheat) or semolina (durum wheat), which is the basic ingredient for all bread and pastry products and for pasta.

[0005] There are also food products composed of whole or crushed wheat grains.

[0006] As an example, we can cite bulgur which is a staple food in the countries of the Middle East itself and which consists essentially of dry wheat grains which are parboiled until they become tender. , then dried and stripped of their outer layers of bran, and finally possibly crushed.

[0007] Furthermore, European patent EP0561092B1 discloses precooked and shelled durum wheat grains, manufactured by implementing the following steps: cleaning, calibration, hydration for subsequent gelatinization of the wheat starch (preheating - hot rest - cooking), drying, cooling, mechanical hulling, expansion heat treatment - preferably rewetting. These pre-cooked durum wheat grains must be cooked in water for at least 10 minutes.

In this technical line, the French patent FR2862891B1 describes a method of manufacturing a product intended for human consumption consisting of durum wheat grains. In a first step, wheat grains are harvested in an immature state (green wheat grains: humidity level between 50 and 70% - albumen sugars not yet completely transformed into starch [raffinose<0.4% dry/dry versus 1.2% dry/dry mature wheat]- milky or pasty appearance). A pretreatment of freezing at - 20°C or heating at 200 - 400°C for 1 to 10 minutes until a water content = 10 - 55%, can follow the harvest of immature grains. The glumes and lemmas of the wheat grains are then removed by threshing or brushing. The wheat grains are then brought to humidity levels of between 67 and 74% by bringing their temperature between -80 and 0°C and/or between 70°C and 200°C for 1-10 min and/or 10 -120 mins.

These wheat grains can be eaten cold or hot after cooking in a microwave oven or over low heat.

Their sugar content (% s/s) is given in Table 1 below.

[0009] [Table 1]: sugar content of immature durum wheat grains

[0010] Wheat grains according to FR2862891 B1 always include starch

"popped", most often extrinsically sweet, and usually intended to be eaten at breakfast. These puffed wheat grains lack crispness and have a floury taste. This requires the exogenous addition of sugars.

[0012] The article Genes Genet. System (2006) 81, p. 361-365, Nakamura et al "Sweat wheat" which describes a sweet wheat comprising a mutation of the sslla gene rendering it deficient, which results in a high amylose (HA) phenotypic characteristic of wheat, and a mutation of the gbssl gene rendering it deficient, resulting in a Waxy (Wx) phenotypic trait without amyloidosis. This double mutated SW wheat is characterized by a high milling yield (>50%) and by an endosperm with a high maltose and sucrose content. This article refers to juices extracted from the grains of double mutated wheat, to flour made from these grains of double mutated wheat and to breads obtained from this flour. WO03/041490A2 relates to methods for producing and identifying different isoamylase genes from individual genomes of a polyploid plant. These methods make it possible to produce, by screening, mutation and selection strategies, polyploid plants lacking a functional isoamylase gene for each plant genome. These plants have high levels of free sugars (such as glucose). Furthermore, D2 discloses numerous isoamylase genes (three isoforms), proteins and cDNA sequences from different species of polyploid plants including hexaploid wheat. There is no mention of low milling yields for the wheats described in this PCT patent application.

Objectives of the invention

In this context, the invention aims to satisfy at least one of the following objectives:

[0014] - providing a new food product based on processed wheat grains with low milling yield.

[0015] - provide a new food product based on processed wheat grains rich in soluble sugars and more particularly sucrose, the reference sugar in sweetening power.

[0016] - providing a new food product based on processed wheat grains rich in sugars and without added sugars;

[0017] - providing a new food product based on processed wheat grains rich in fibre;

- provide a new food product based on processed wheat grains with improved organoleptic properties; - provide a new food product based on processed wheat grains with masking properties of undesirable tastes;

- provide a new food product based on processed wheat grains consisting of an integral flour of high milling value, high baking value, high nutritional value and high commercial value. [0021] to provide a transformation process making it possible to obtain the food product referred to in at least one of the above objectives, in an industrial, simple and economical way. Brief description of the invention

The invention satisfies at least one of the above objectives and relates, according to a first aspect, to a food product comprising processed wheat grains having a Milling Yield RM produced according to standard NF EN ISO 27971, expressed in % by mass of dry matter relative to whole wheat grains, such as, in increasing order of preference:

- 0.5 ≤ RM ≤ 10;

- 1 ≤ MR ≤ 5;

- 1.5≤RM≤3. dietary plan due to the presence of fiber from the pericarp (bran) and the valorization of this wheat in integral flour.

[0024] These processed wheat grains, in particular in the form of integral flours, constitute very interesting food ingredients, even at low percentages of incorporation, for more elaborate food compositions, precisely because of the qualities they carry. , in particular, presence of high sucrose within a complex matrix and high fiber content.

[0025] This new product can also benefit from the advantageous commercial claim: “without added sugars” or “reduced in added sugars”.

The nutritional qualities of these new processed wheat grains relate in particular to the presence:

- sucrose in a concentration 3 to 20 times, preferably 6 to 12 times greater, for example, than in the wheat grains known until then; - and fibers in a concentration 2 to 3 times higher, for example, than in the wheat grains known until now.

[0027] These new processed wheat grains also offer significant organoleptic benefits, in particular by allowing the bitterness of the bran to be smoothed and a softer texture for “whole” bread products. They thus bring a gourmet flavor to the food products, e.g. industrial ones, in which they are incorporated.

These new processed wheat grains have a low milling yield reflecting a reduced starch content or even the absence of starch. These new grains of processed wheat form whole grain products without a mealy and crispy taste.

[0029] Moreover, these grains of wheat perform remarkably well with regard to their agricultural production. They are characterized by very good germination and the formation of very beautiful plants.

The new food product according to the invention is also unique in that these wheat grains have at least one of the following characteristics:

(-2.1-) Fiber Content (FI), expressed as % by mass of material such as, relative to whole wheat grains, such as, in increasing order of preference:

-10≤TF≤45;

-15≤TF≤40;

- 20 ≤ TF ≤ 30;

(-2.2-) Ash content (TC), expressed in % by mass of material such as , relative to whole wheat grains, such as, in increasing order of preference:

- 1.6 ≤ CT ≤ 3;

- 1.8 ≤ CT ≤ 2.8;

- 2 ≤ CT ≤ 2.5;

(-2.3-) Protein content (TP), expressed as % by mass of material such as relative to whole wheat grains, such as, in ascending order of preference:

- 13≤TP≤35;

- 15 ≤ TP ≤ 30;

- 18 ≤ TP ≤ 25;

(-2.4-) Total Sugar Content (TST), expressed in %, by mass of material such as in relation to whole wheat grains, such as, in increasing order of preference:

- 1.5 ≤ TST ≤ 40;

- 2 ≤ TST ≤ 30;

- 2.5≤TST≤15;

(-2.5-) Sucrose content (ST), expressed as % by mass of matter such as, relative to whole wheat grains, such as, in increasing order of preference:

- 4 ≤ ST ≤ 30;

- 5 ≤ ST ≤ 20;

- 6 ≤ TS ≤ 16;

- 7 ≤ TS ≤ 15; - 8 ≤ TS ≤ 14.

(-2.6-) phenotypic appearance of the crumpled type.

The new food product according to the invention has the particularity that the wheat grains which it comprises, have a deficiency in at least one enzymatic activity involved in the biosynthesis and/or the structuring of amylopectin which opposes the formation of starch granules, this deficiency preferably being this deficiency preferably resulting from:

- a deficiency, total or partial, in the activity per se of at least one enzyme involved in the biosynthesis and/or the structuring of amylopectin, which interferes with the formation of starch granules, and/or

- a deficiency, total or partial, in the expression of at least one gene coding for this

(these) enzyme(s).

Preferably, the processed wheat grains of the food product according to the invention present on at least one of their genomes, preferably on at least 2 of their genomes, and, more preferably still, on all their genomes, at least least one mutation of the iso-1 gene corresponding to the protein/enzyme ISO-1 and/or of the sh-2 gene corresponding to SH-2 corresponding to the protein/enzyme SH-2, this mutation or these mutations leading to a deficiency, total or partial, of the expression of the iso-1 gene and/or a deficiency, total or partial, of the expression of the sh-2 gene corresponding to SH-2, respectively.

According to a preferred embodiment of the invention, the deficiency results from a genetic variation identified within a Tilling population by means of specific markers. According to alternatives of this privileged mode, the deficiency in enzyme(s) necessary for the formation of the starch granule can also be obtained in particular:

- by identifying mutants deficient in enzymes necessary for the formation of the starch granule, - in collections of soft or durum wheat that can be obtained by transgenesis, mutagenesis,

- by new plant selection techniques (New Breeding Techniques),

- or by implementing the “Clustered Regularly Interspaced Short” technology Palindromie Repeats” (“Short clustered and regularly spaced palindromic repeats”), more commonly referred to as CRISPR.

It is understood that individual mutants identified in one collection can be combined with other mutants from other collections to achieve the desired phenotype.

In a first remarkable embodiment, the food product according to the invention comprises wheat grains which are immature and which have a humidity H°, expressed in% by mass, such that, in increasing order of preference:

- 16 ≤ H° ≤ 80;

- 45 ≤ H° ≤ 75;

- 55 ≤ H° ≤ 70.

In this first embodiment, the processing of immature wheat grains consists of at least one of the following treatments: canning, freezing, drying, steaming, heating to a temperature between 150 and 300° C., preferably between 170 and 250° C. at a heating rate chosen to allow the splitting of the grains (popping).

In a second remarkable embodiment, the food product according to the invention comprises wheat grains which are mature and which have a measured humidity H°, expressed in% by mass, such that, in ascending order of preference :

- 2 ≤ H° ≤ 16;

- 5 ≤ H° ≤ 16;

- 8 ≤ H° < 15 .

In this second embodiment, and according to a first possibility, the transformation of mature wheat grains consists of at least one of the following treatments:

(a) heating to a temperature of between 150 and 300°C, preferably between 170 and 250°C at a heating rate chosen to allow the splitting of the grains (popping);

(b) puffing;

(c) Toasting;

(d) Frying at a temperature of between 160 and 200° C., preferably of the order of 180° C. for 5 to 60 s, preferably 10 to 30 s;

(e) Flaking; (f) Soaking in water;

(g) Steaming;

(h) Boiling;

(i) Extrusion. In this second embodiment, and according to a second possibility, the processing of mature wheat grains consists of grinding to obtain semolina or integral flour, and a possible heat treatment of this semolina or this flour.

[0040] According to a ^second of its aspects, the invention relates to bread products obtained from flour or semolina resulting from the processing referred to in paragraph [0040].

[0041] According to a ^third of its aspects, the invention relates to pastry products obtained from flour or semolina resulting from the processing referred to in paragraph [0040]. [0042] According to a ^fourth of its aspects, the invention relates to a sourdough, in particular a liquid sourdough, obtained from flour or semolina resulting from the processing referred to in paragraph [0040].

According to a ^fifth of its aspects, the subject of the invention is a transformation process making it possible to obtain the food product according to the first embodiment referred to above, characterized in that it consists:

-1- to use immature wheat grains whose humidity H°, expressed in % by mass, is such that: 16 < H° ≤ 80;

-2- in subjecting these wheat grains to at least one of the following treatments: canning, freezing, drying preferably at a temperature between 30 and 60° C., preferably between 40 and 50° C., for 5 to 48 hours preferably 20 to 30 hours.

-3- and, possibly, to condition them.

According to a ^6th of its aspects, the subject of the invention is a transformation process making it possible to obtain the food product according to the second embodiment referred to above, characterized in that it consists of:

-1- to implement mature wheat grains whose humidity H °, expressed in% by mass, is such that: 2 ≤ H ° ≤ 16; -2- subjecting these wheat grains to at least one of the following treatments:

(a) heating to a temperature between 150 and 300°C, preferably between 170 and 250°C at a heating rate chosen to allow the expansion of the grains (popping).

(b) Puffing

(c) Toasting

(d) frying at a temperature of between 160 and 200° C., preferably of the order of 180° C. for 5 to 60 s, preferably from 10 to 30 s.

(e) flaking;

(f) soaking in water.

(g) steaming.

(h) extruding.

-3- possibly conditioning them;

-4- and possibly canning them.

According to a ^7th of its aspects, the subject of the invention is a wheat characterized in that it has on at least one of its genomes, preferably on at least 2 of its genomes, and, even more preferentially, on all their genomes, at least one mutation of the iso-1 gene coding for the ISO-1 protein/enzyme and/or of the sh-2 gene coding for the SH-2 protein/enzyme, this mutation or these mutations resulting in a deficiency, total or partial, of the expression of the iso-1 gene and/or a deficiency, total or partial, of the expression of the sh-2 gene corresponding to SH-2, respectively.

Preferably, the wheat according to the invention is characterized in that the mutations are chosen from: ApC-5022; ApaA-2164; ApC-; Apa-1119; ApA-0866; and their combinations.

Brief description of the drawings

This description is made with reference to the appended figures illustrating non-limiting examples of embodiment. In this brief description of the figures and throughout the remainder of this presentation and in the appended figures, "Isowheat" or "Isowheat sweet wheat" designates the wheat used in the food product according to the invention or the wheat according to the invention. and “Apache” or “standard wheat” corresponds to a control wheat which serves as a reference in the examples.

Fig. 1

[0048] [Fig. 1] Figure 1 is a photograph of the crumpled mature wheat grains constituting the food product according to the invention.

Fig. 2

[0049] [Fig. 2] Figure 2 is a diagram showing the position of the genetic variation G→A (ApC-5022) on the iso-1 gene located on chromosome 7 AS of wheat Triticum aestivum of example 1.

Fig. 3

[0050] [Fig. 3] Figure 3 is a diagram showing the position of the genetic variations G→A (ApA-2164) and G→A (ApA-1119) on the iso-1 gene, located on the chromosome

7BS of Triticum aestivum wheat from Example 1.

Fig. 4

[0051] [Fig. 4] Figure 4 is a diagram showing the position of the genetic variation G→A (ApA-0866) on the iso-1 gene, located on the 7DS chromosome of wheat Triticum aestivum of example 1.

Fig. 5

[0052] [Fig. 5] FIG. 5 represents the projection of the position of each genetic variation identified in the sequences of paragraphs [0244] [0245] [0246] below, on the reference protein sequence of isoamylase lso-1 SEQ ID N °1, these projections being predicted from the sequences SEQ ID No. 2 to 5 of the iso-1 7AS, 7BS and 7DS gene of the wheat Isowheat Triticum aestivum according to the invention (example 1).

Fig. 6

[0053] [Fig. 6] Figure 6 consists of photographs representing a plot of Isowheat wheat, ears of immature Isowheat wheat grains (70.2% humidity) and reference Apache wheat (56.1% humidity) harvested at the same date (June 17, 2020). [Example 1].

Fig. 7

[0054] [Fig. 7] Figure 7 consists of photographs of immature ears of Isowheat wheat (sweet wheat) and Apache wheat (standard wheat) harvested on June 17, 2020, and their grains after threshing. [Example 1]. Fig. 8

[0055] [Fig. 8] Figure 8 consists of photographs of immature ears of Isowheat wheat (sweet wheat) and Apache wheat (standard wheat) harvested on June 23, 2020, and their grains after threshing. [Example 1]. Fig. 9

[0056] [Fig. 9] Figure 9 shows immature Isowheat and Apache grains harvested on June 23, 2020 after steaming for 20 minutes in a kitchen steamer. [Example 2].

Fig. 10 [0057] [Fig. 10] Figure 10 consists of photographs of immature ears of wheat

Isowheat and Apache harvested on July ¹ , 2020, and their grains after threshing. [Example 2] ·

Fig. 11

[0058] [Fig. 11] Figure 11 shows immature Isowheat and Apache grains harvested July ¹ , 2020 after steaming for 25 minutes in a kitchen steamer. [Example 2].

Fig. 12

[0059] [Fig. 12] Figure 12 shows Isowheat and Apache grains harvested on two different dates, June 23, 2020 (stage 1) and July ¹ , 2020 (stage 2). [Example 2]. Fig. 13

[0060] [Fig. 13] Figure 13 shows photos of immature Isowheat and Apache beans harvested June 17, 2020, June 23, 2020, and July ¹ , 2020 canned. [Example 2]

Fig. 14

[0061] [Fig. 14] Figure 14 shows native and popped Isowheat grains for 35, 40, and 45 seconds. [Example 2].

Fig. 15

[0062] [Fig. 15] Figure 15 shows Isowheat grains fried in oil for 30 seconds at 180°C. [Example 2].

Fig. 16

[0063] [Fig. 16] Figure 16 shows the diagram of the puffing process. [Example 2]. Fig. 17

[0064] [Fig. 17] Figure 17 shows native and puffed Apache and Isowheat grains. [Example 2] ·

A: Native Apache grains, B: Apache grains puffed at 10 bars and for 10 seconds, C: Apache grains puffed at 21 bars and for 25 seconds, D: Native Isowheat grains, E: Isowheat grains puffed at 10 bars and for 10 seconds , F: Isowheat grains puffed at 21 bars and for 25 seconds, G: Isowheat grains puffed at 12 bars and for 12 seconds

Fig. 18

[0065] [Fig. 18] Figure 18 is a photograph showing the total flours from Chopin Dubois milling of Apache (reference) and Isowheat wheat grains from Example 6.

Fig. 19

[0066] [Fig. 19] Figure 19 is a photograph showing the sounds from the Chopin Dubois milling of Apache (reference) and Isowheat wheat grains from example 6.

Fig. 20 [0067] [Fig. 20] Figure 20 is a photograph showing the remoulding flours from the Chopin Dubois milling of Apache (reference) and Isowheat wheat grains from Example 6.

Detailed description of the invention

In general, any singular designates a plural and vice versa. [0069] Food product: General

The food product according to the invention is based on processed wheat grains.

[0070] "Transformed wheat grains" are, for example, within the meaning of the invention, whole wheat grains no longer having the capacity to germinate and develop into a plant, harvested in the mature state or the immature state, and having undergone at least one transformation, preferably industrial, chosen from the group comprising - ideally composed of -:

. grinding,

. beading (removal of at least part of the envelope and/of at least part of the germ of the grain of wheat), . grind,

. driving, . crushing,

. cooking (in particular steam and/or water and/or dry, and/or in an oven),

. extrusion,

. toasting. blowing "popping",

. burst "puffing",

. freezing,

. canning,

. pasteurization, . conditioning,

. and their combinations.

[0071] Within the meaning of the invention, “transformation” of the grains designates, for example, an industrial transformation or a physiological transformation followed by an industrial transformation. A physiological transformation can be, for example, that induced by the sowing, cultivation and harvesting of grains.

An industrial transformation can be, for example, at least one of those listed in the previous paragraph.

In the case where the grains are harvested in the immature state, the transformation is advantageously a physiological and industrial transformation.

[0073] By “whole wheat grains”, is meant, for example, according to the invention, wheat grains comprising the brush, the envelopes (pericarp, seminal integument and hyaline band), the albumen (aleurone layer, albumen) and the germ or embryo (coleoptile, gemmule, embryonic axis, radicle, coleorhize). By "immature wheat grains" is meant, for example, according to the invention:

^* wheat grains which have a humidity H°, expressed in % by mass, such as:

16 < H° ≤ 80;

^* and/or grains which ferment under ambient conditions (eg 20°C, 760 mm Hg) and thus become unfit for consumption. By "mature wheat grains" is meant, for example, according to the invention:

^* wheat grains which have a humidity H°, expressed in % by mass, such as:

2 < H° ≤ 16; ^* and/or grains that can be stored under ambient conditions (for example 20°C, 760 mm Hg), without fermenting and therefore without risk of becoming and thus becoming unfit for consumption.

By “dry wheat grains”, is meant, for example, according to the invention, wheat grains which have a humidity H°, expressed in% by mass, less than or equal to 16.

The mass percentages used in the present presentation are expressed relative to the total mass of the whole wheat grains considered, including the water included in these wheat grains. In the event that percentages by weight on a dry basis are used, the mention “on a dry basis” will be expressly mentioned. The humidity H° (or water content) is measured according to standard NF EN ISO 712: 2009.

This humidity H° between 2 and 16% by mass is the humidity of the grains of wheat harvested in the mature state after desiccation of the ear on the plant or of the grains of wheat harvested in the immature state and dried up to H ° between 2 and 16% by mass.

The wheat grains of the food product according to the invention have a miller yield RM achieved with dry wheat grains, according to standard NF EN ISO 27971 and expressed in % by mass relative to whole wheat grains, such as , in ascending order of preference:

- 0.5 ≤ RM ≤ 10;

- 1 ≤ MR ≤ 5; - 1.5 ≤ RM ≤ 3.

This miller yield characteristic RM applies to dry wheat grains or to mature wheat grains within the meaning of the invention.

Another remarkable and potential phenotypic characteristic of the wheat grains specific to the food product according to the invention is that these wheat grains are free or almost free of starch, and more precisely of starch granules, and this , whatever the humidity H of these grains.

Without wishing to be bound by theory, it is possible that the total carbohydrate content -2.4- and/or the sucrose content -2.5- characteristic(s) of the wheat grains of the food product according to the invention, is (in) t linked to the absence or virtual absence of starch in these wheat grains.

[0082] This absence or this starch deficiency can be demonstrated by an observation such as the person skilled in the art can do, under an optical microscope, for example magnification 40 times, of a ground wheat grain suspended, optionally stained with iodine. This deficiency is highlighted in Example 6 below, by a milling yield of less than 3% for Isowheat wheat, instead of 54% in the standard signifying a virtual absence of white flour due to a deficiency in starch which is its main constituent (70%). The wheat grains of the food product according to the invention are also distinguished in that they have at least one of the characteristics -2.1-2.2-2.3-2.4-2.5-2.6- mentioned above.

The characteristic (-2.1-) of Fiber Content (TF), between 10 and 45% by mass relative to whole wheat grains, corresponds to a fiber concentration measured according to the AOAC 985.29 standard.

Thus, the TF can be close to double or more than in Apache-type reference wheat grains.

The characteristic (-2.2-) of ash content (TC) of between 1.6 and 2.5% by mass relative to whole wheat grains corresponds to an ash concentration measured according to the ISO 2171 standard: 2007.

The characteristic (-2.3-) of Protein Content (TP) of between 15 and 30% by mass relative to whole wheat grains, corresponds to a protein concentration measured according to standard NF EN ISO 16634 - V18- 130PR - February 2004.

The characteristic (-2.4-) of Total Sugar Content (TST) between 1.5 and 40% by mass corresponds to a percentage of total sugars calculated by summing the contents of the following sugars: glucose, fructose , maltose, lactose and sucrose measured according to the sugar assay method by high performance anion exchange chromatography coupled with detection by pulsed amperometry (HPAEC-PAD). For this sugar assay method, refer to the following sugar assay reference documents: glucose, fructose, maltose, lactose and sucrose:

❖ AOAC 980.13 Method - Fructose, Glucose, Lactose, Maltose, and Sucrose in Milk Chocolate - Liquid Chromatography Method - Final Action,

❖ AOAC Method 982.14 - Glucose, Fructose, Sucrose, and Maltose in Presweetened Cereals - Liquid Chromatography Method - First Action 1982 -

Final Action 1983,

❖ Labo-Stat, Validation guide for analysis methods, Max Feinberg,

LAVOISIER 2009. The characteristic (-2.5-) of sucrose content (ST) of between 4 and 30% by weight corresponds to a sucrose concentration measured in the grains or in the immature grains.

According to a reference method, the assay is carried out according to the method for assaying sugars by high performance anion exchange chromatography coupled with detection by pulsed amperometry (HPAEC-PAD).

For the assay method, refer to the reference documents for the assay of sugars: glucose, fructose, maltose, lactose and sucrose:

^* AOAC Method 980.13 - Fructose, Glucose, Lactose, Maltose, and Sucrose in Milk Chocolate - Liquid Chromatography Method - Final Action,

^* AOAC Method 982.14 - Glucose, Fructose, Sucrose, and Maltose in Presweetened Cereals - Liquid Chromatography Method - First Action 1982 - Final Action 1983,

^* Labo-Stat, Validation guide for analysis methods, Max Feinberg, LAVOISIER 2009.

The sugars present in the test sample are extracted with vigorous stirring, after addition of an internal standard, with a water/ethanol mixture at a temperature of 85°C. The extract is then clarified by Carrez reagents, then filtered and finally diluted before being injected into the high-performance anion exchange chromatography system coupled with detection by pulsed amperometry. Sugars are quantified by comparing the areas of the peaks with those of an internal or external calibration curve established with standard solutions.

The characteristic (-2.6-): crumpled phenotype, is specific to mature wheat grains or to dry wheat grains, within the meaning of the invention. Characteristic -2.6- is optional but nevertheless interesting. This is a phenotypic characteristic, relating to the wrinkled "shrunken" appearance of the wheat grains shown in the appended figure 1.

Preferably, the wheat grains of the product according to the invention have a deficiency in at least one enzymatic activity involved in the biosynthesis and/or the structuring of amylopectin, which thwarts the formation of starch granules.

This deficiency, total or partial, of this or these enzymatic activities may result from:

- a deficiency, total or partial, in the activity per se of at least one enzyme involved in the biosynthesis and/or the structuring of amylopectin, which interferes with the formation of starch granules, and or

- a deficiency, total or partial, in the expression of at least one gene coding for this (these) enzyme(s).

According to a preferred embodiment, the wheat grains of the food product according to the invention present on at least one of their genomes, preferably on at least 2 of their genomes, and, more preferably still, on all their genomes, at least one mutation in the sequence of the iso-1 gene coding for the ISO-1 protein/enzyme and/or in the sequence of the sh-2 gene coding for the SH-2 protein/enzyme, this mutation or these mutations causing a deficiency, total or partial, of the expression of the iso-1 gene and/or a deficiency, total or partial, of the expression of the sh-2 gene.

This deficiency in the expression of genes is linked to the alteration of their sequence which is likely to cause transcription and/or translation anomalies. The mutation in the coding sequence of the gene can also lead to a total or partial deficiency of the enzymatic activity. [0094] The "deficiency" means, in the sense of the invention, e.g. in relation to a reference wheat which can be for example an APACHE wheat

According to a first embodiment of the invention, the deficient enzymatic activity results:

- a total or partial deficiency in isoamylase 1 (ISO-1) enzymatic activity; and or

- a deficiency, total or partial, in at least one protein corresponding to ISO-1; and or

- a deficiency, total or partial, in the expression of the iso-1 gene coding for the ISO-1 protein. According to a second embodiment of the invention, the deficient enzymatic activity results:

- a deficiency, total or partial, of ADP-glucopyrophosphorylase-2 or shrunken-2 (SH-2) enzymatic activity; and or

- a deficiency, total or partial, in at least one protein corresponding to SH-2; and or

- a total or partial deficiency in the expression of the sh-2 gene coding for the SH-2 protein.

According to a third embodiment of the invention, the deficient enzymatic activity results: - a deficiency, total or partial, of isoamylase 1 (ISO-1) enzymatic activity and/ of ADP-glucopyrophosphorylase-2 or shrunken-2 (SH-2) enzymatic activity; and or

- a deficiency, total or partial, in at least one protein corresponding to ISO-1 and in at least one protein corresponding to SH-2; and/or - a total or partial deficiency in the expression of the iso-1 gene coding for the ISO-1 protein and in the expression of the sh-2 gene coding for the SH-2 protein.

According to an advantageous possibility of the invention, this deficiency is obtained by genetic variation identified in at least one of the genomes A, B, D of wheat (genus Triticum), preferably all three, within at at least one tilling population, for example a wheat population of the APACHE line, by means of specific markers.

According to a remarkable characteristic of the invention, the genetic variation is identified according to the following conventional steps for identifying genetic variation: From the DNA extracted from the individuals of the tilling population or populations, amplicons are generated at using pairs of specific primers, preferably genome (A, B, D) specific. Amplicons are sequenced and analyzed to select genetic variations of interest. Genotyping and selection using specific markers of individuals from all or part of the pools is carried out to identify the M1 mutant individuals include the selected genetic variations;

The genotyping and selection process is carried out on individual M2 mutant offspring. Individuals exhibiting at least one genetic variation on at least one genome are crossed to generate F1 individuals exhibiting double genetic variations. A series of crosses is carried out to generate an F2 progeny comprising double homozygous genetic variations and then, from this, to obtain a progeny comprising individuals presenting triple homozygous genetic variations.

The genetic variations of interest are those which make it possible to obtain a deficiency, total or partial, of the expression of the iso-1 gene and/or a deficiency, total or partial, of the expression of the gene sh-2 or a a deficiency, total or partial, of the enzymatic activity of the proteins encoded by these genes.

[0100] Food product: Example of making wheat

In a preferred embodiment of the invention, the wheat grains are derived from wheat plants exhibiting genetic variations in the iso-1 gene called Isowheat. The genetic variations in the iso-1 gene are advantageously chosen from: ApC-5022; ApaA-2164; Apa-1119; ApA-0866; and their combinations.

ApC-5022; ApaC-2164; Apa-1119; ApA-0866 are defined by their respective context sequence, SEQID No. 2, 3, 4, 5. Each context sequence can be compared to the reference sequences SEQ ID No. 6, 7 and 8 of the iso-17AS gene of genome A , iso-1 7BS gene from the B genome and iso-1 7DS gene from the D genome of Triticum aestivum, in order to position the genetic variation in the gene sequence.

The sequence of the iso-1 gene of the wheat grains transformed according to the invention also encompasses any similar gene sequence which exhibits, in addition to the genetic variations according to the invention, genetic variations other than those defined according to the invention. This includes sequences homologous to the reference sequences SEQ ID Nos. 6, 7 and 8, knowing that the term "homologous" preferably means nucleic acid sequences having at least 90% identity, and preferably at least 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity with the reference sequences SEQ ID Nos. 6, 7 and 8.

[0104] FIG. 5 represents the projection of the position of each genetic variation identified in the sequences [0244] [0245] [0246] and in table 2 below, on the reference protein sequence of isoamylase ISO-1 SEQ ID No. 1 predicted from the sequences SEQ ID No. 6, 7 and 8 of the iso-1 7AS, 7BS and 7DS genes of Isowheat wheat according to the invention.

The predicted ISO-1 protein sequence of wheat grains transformed according to the invention also encompasses any similar protein sequence in which the locations of the mutations described above would be identical, but the nature of the mutations would be different. This also encompasses the homologs of this ISO-1 protein sequence described above, knowing that by "homolog" is meant, preferably, protein sequences having at least 90% identity, and preferentially at least 91%, 92% , 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity with the reference sequence SEQ ID No. 1.

[0106] Percent identity is determined by calculating the number of positions that match on aligned amino acid sequences, dividing the number of positions that match by the total number of amino acids, and multiplying by 100 A match position refers to a position for which identical amino acids are present at the same position in the alignments of amino acid sequences. For example, amino acid sequences can be aligned using CD-hit (http://weizhongli-lab.org/cd-hit/).

[0107] Food product: first embodiment with immature wheat grains

In this first embodiment, the processed wheat grains are immature, which translates into a humidity H° greater than 16%, preferably between 16 and 80%.

[0109] The transformations that can be made on these immature wheat grains are in particular canning, freezing, drying, steaming, popping... [0110] Within the meaning of the invention, popping is, for example, subjecting a grain of whole dry wheat set in motion by air to a temperature of 240° C. for 30 seconds. Under the effect of heat, the water contained in the wheat grain passes into vapor form creating sufficient internal pressure to expand the structure of the wheat grain without exploding (not like puffing). Freezing can, for example, take place just after (e.g. 1-5 hours) threshing of the harvested wheat grains, at a temperature between -50 and -20°C (e.g. -30°C) for a period of between 1 to 5 hours (e.g. 2h).

The grains can be dried in an oven, at a temperature of between 30 and 60° C., for example 45° C., for 12 to 48 hours, for example 24 hours. After drying, wheat grains remarkably exhibit a wrinkled “shrunken” type phenotypic appearance [Figure 1] similar to that of mature grains. The absence of starch does not allow the wheat grain to have a “full” structure due to a lack of material that cannot completely fill the grain envelope.

Advantageously, the drying of the grains is complete when their weight no longer evolves while being placed in an oven between 30 and 60°C, typically their humidity is then between 5 and 16%.

[0113] According to another possibility, the steam cooking of the immature grains is carried out using a steam cooker with cooking for 20 minutes

According to another possibility, immature grains dried in an oven so that their humidity H is between 5 and 16% are subjected to heating under air flow at a temperature between 150 and 300° C., preferably between 170 and 250°C at a heating and air flow rate chosen to allow the bursting of the grains (popping).

[0115] Food product: second embodiment with mature wheat grains

In this second embodiment, the processed wheat grains are mature, which results in a humidity H greater than or equal to 2% and less than 16%.

[0117] A phenotypic characteristic of these mature wheat grains is their wrinkled appearance, as defined above and shown in the appended FIG.

[0118] The possibilities of transformation are the treatments (a)(b)(c)(d)(e)(f)(g)(h)(i) which are mentioned above and which are detailed below . (a) Popping: Mature grains are subjected in centrifugal motion by a flow of air, to heating at a temperature of between 150 and 300° C., preferably between 170 and 250° C. at a heating rate chosen to allow the splitting of the grains.

b) Puffing: The wheat grains are sent to an expansion chamber heated to a temperature of 95°C. After introducing the grains, the chamber is closed and put under pressure, by injecting steam, between 10 and 25 bars. After a residence time of 10 to 30 seconds, the chamber is opened, releasing the grains which, under the effect of the depression, will expand (puffer).

[0121] (c) Toasting: Lees grains are placed in a frying pan and heated by stirring them on a thermostat 8 induction hob, until they take on a filled form.

[0122] (d) Frying: The dry grains placed in a tight-mesh strainer are immersed in a frying bath at a temperature of 180° C. for 15 seconds.

(e): Flaking: The whole seed is steamed for 20-30 minutes. Once cooked the seeds, at a humidity of 16 to 18% are passed through a roller to be reduced to a thickness of less than 1 mm. To preserve their shelf life, the flakes are stabilized by drying in the oven or in the air until a humidity of 3% is reached.

(f) Soaking in water: The grains are placed in a container and covered with water at room temperature, at 40°C or at 4°C, the Isowheat grains rehydrate, swell and acquire a crunchy, non-mushy texture.

[0123] (g) Steaming: The wheat grains are placed in a kitchen steamer, SEB® minicompact, and subjected to steam for a period varying from 10 to 90 minutes, allowing rehydration and cooking of the grain. The texture of the cooked grain is crunchy and not pasty.

[0124] (h) Boiling: Wheat grains are placed in a saucepan and covered with water. The water is brought to a boil and the grains are cooked for a period varying from 10 to 90 minutes and have a filled appearance and a crunchy texture, not pasty.

[0125] (i) Extrusion: The introduction of ground Isowheat wheat grains into a single-screw or twin-screw extruder makes it possible to obtain billets with a sweet taste.

[0126] Food product: third embodiment with immature & mature wheat grains [0127] In this 3 ^rd embodiment, mixtures of immature wheat grains processed according to the first embodiment and wheat grains are envisaged. mature wheat processed according to the 2nd ^embodiment .

[0128] Food product: 4 ^th embodiment with mature and/or immature wheat grains reduced to a humidity of less than 20% [0129] In this embodiment, the wheat grains according to the invention, mature and/or immature eggs reduced to a humidity of less than 20% are transformed by grinding to obtain semolina, preferably integral/complete semolina, or integral/complete flour.

These transformation processes are conventional processes such as those commonly used in grinding operations in the cereal food industry using e.g. hammer mills.

[0130] Advantageously, semolina or wholemeal flour can, conventionally, be heat-treated, to increase its shelf life by destroying microorganisms and by inactivating endogenous enzymes, with the possibility of modifying the rheological parameters of the flour (viscosity) . For this, the flour must be subjected to a heat treatment at a temperature above 60°C, by steaming, hydrothermal treatment, extrusion cooking or by drying cylinders for example.

[0131] Food compositions obtained from the food product according to the invention [0132] The applications of the Wheat Grains Processed According to the Invention (GBTSI) can be, among others:

- Whole GBTSI as a garnish on breads or pastries, - Whole GBTSI as ingredients incorporated into bread or pastry products as a marking agent or crisping agent,

- GBTSI in cans,

- GBTSI in cooked dishes, - GBTSI as products to be cooked in water such as pasta,

- expanded but not burst whole GBTSI (popped type) as a nutritional/health/dietetic product,

- Expanded whole GBTSIs but not popped (popped type), by frying as a tasty product, - Expanded whole GBTSIs as a snack as is or in a salty or sweet coating,

- whole GBTSI as ready-to-eat cereals: e.g. breakfast cereals,

- whole GBTSI as ingredients in cereal bars,

- Flour/semolina, in particular integral or complete flour/semolina, from GBTSI,

- Bread or pastry product made from this flour/semolina from GBTSI,

- Leaven, in particular liquid leaven, obtained from this flour/semolina by mixing with water, for example at 30-35° C. from GBTSI. Such leaven can be prepared by carrying out several successive fermentations at temperatures between 25 and 35° C., for several hours (8 hours-24 hours). At each fermentation, a new flour/water mixture is added to the mixture resulting from the previous fermentation. The sourdough obtained is stored in the cold.

The advantages induced by the food product according to the invention in these compositions are in particular:

^* a sweetening power allowing the use of the nutritional claim "no added sugar" or "less X% sugar" or of the "Nutri-Score" type in complete finished products, with the same sweetness and less sugar;

^* an enrichment in fibers with strong nutritional and dietary virtues and which can enable a product to achieve the nutritional claim “source of fibre” or “rich in fibre”; * a particularly interesting technological auxiliary role in terms of water absorption, texture, fermentability, in particular in bread products and in particular in sourdough bread;

^* an organoleptic enrichment providing a less rough tasting sensation, a smoothing of the bitterness of the sounds and a softer texture on the complete products, a masking effect which can be interesting in pastry, better acceptance by the consumer of products that are nutritionally interesting but not very palatable for the consumer.

Examples

[01341 Example 1: OBTAINING DEFICIENT WHEAT

A tilling population of 5351 individuals from the Apache lineage was used. 56 DNA pools were constructed from the DNA extracted from the 5351 individuals in the population. A second tilling population of 912 individuals from the Arche lineage was used. Ten DNA pools were constructed from the DNAs extracted from the 912 individuals. Amplicons were generated from the 66 pools using genomic primers located in exons 2 to 18 of the iso-1 gene of the Chinese Spring reference sequences (Triticum aestivum L.) SEQ ID N°6, 7 or 8 Amplicons were sequenced by Illumina Solexa sequencing. Of the 106 genetic variations or mutations selected, 46 were confirmed by genotyping of the M2 families. Crosses are carried out in order to obtain wheat plants comprising two genetic variations each on two different genomes (double mutant) or three genetic variations each on three different genomes (triple mutant). Table 2 below represents the genotyping results of the triple mutants thus selected and exhibiting the ISOWHEAT phenotype. The IS017-001 and IS017-002 lines are used for the experiments described in the following examples.

Table 2 below summarizes the genotyping of the two lines IS017-001 and IS017-002 with respect to each of the 4 ApC-5022 genetic variations; ApaA-2164; Apa-1119; ApA-0866. The position of each variation on the corresponding iso-1 SEQ ID 6, 7 or 8 gene sequence is given in FIGS. 2 to 5.

[0139] [Table 2]: ISQ17-001 and ISQ17-002 genotyping [0140] Example 2: HARVEST OF IMMATURE GRAINS

The wheat grains according to the invention, hereinafter referred to as sweet wheat or Isowheat, used in this example were obtained by the method described above. The standard reference wheats are APACHE A commercial variety of soft wheat suitable for bread-making, produced and marketed by LIMAGRAIN LG Semences.

[0142] The Isowheat and Apache grains are produced in test plots with the cultivation route practiced by the person skilled in the art (the breeder) in Limagne.

[0143] Samples at different stages of immature ears are taken in order to be able to evaluate them.

[0144] ^1st direct debit: June 17, 2020

[0145] First sampling of immature wheat from Apache (standard) and Isowheat reference wheat ears (see photos in appended FIG. 4).

The stage of immaturity of this sample, stage 1, corresponds to a humidity H for Isowheat of 70.2% and for Apache: 56.1%. The test plot shows Isowheat sweet wheat plants and ears at a slightly earlier stage than the Apache standard as suggested by a greener color and higher humidity.

The threshing of Isowheat and standard Apache sweet wheat ears is carried out using a WINTERSTEIGER LD 210 type laboratory threshing machine. The threshing speed is set to 7 for Apache ear threshing and 6 for Isowheat and fan speed set to 80% for both ear types. These parameters are adjusted according to the samples to limit the breakage of immature grains as much as possible. With these conditions there were no visible broken grains.

Starting from 4.7 kg of Isowheat wheat ears, 3.7 kg of grains were obtained and from 3.3 kg of Apache standard wheat ears 2.4 kg of grains. See attached figure 6.

After threshing, the grains were subjected to freezing, using a CONS-0010 PANEM® BG30-F44 2000-508A freezer deep freezer at a temperature of -30°C for 1 hour, harvested around 10:45 a.m. and frozen at 12:45 p.m., before storage at -18°C. Grains were also left to dry at 45° C., using an oven, until completely dry (approximately 24 hours).

At the taste level, the native Isowheat wheat grains clearly appear sweeter than the native Apache wheat grains. [0151] ^2nd direct debit: June 23, 2020

The stage of immaturity of this sample, stage 2, corresponds to a humidity H for Isowheat of 69.2% and for Apache of 52%.

The harvest of immature ears gave 5.2 kg for Apache and 5.9 kg for Isowheat for a quantity of grains of 4.12 and 4.4 kg respectively.

[0153] During the threshing carried out under the same conditions as for the June 17, 2020 sampling, the Isowheat stalks are noted as being particularly sticky. Isowheat grain is greener, at an earlier stage, larger with nicer ears than standard Apache (see attached figure 3). [0154] Isowheat native immature grain compared to Apache has a distinctly sweeter taste.

[0155] After steaming, using a SEB® mini-compact kitchen steamer, for 20 minutes, the sweet wheat grains are judged to be sweeter, crunchier and better than the standard Apache grains.

Figure 9 shows these soft and standard immature beans harvested on June 23 after steaming for 20 minutes.

As in the previous stage, the grains are frozen and a sample is left to dry at 45° C. for 24 hours.

[0158] 3rd ^direct debit: July 01, 2020

The stage of immaturity of this sample, stage 3, corresponds to a humidity H for Isowheat of 61.8% and for Apache of 41.9%.

[0160] The Isowheat grains compared to the Apache standard are at a less advanced stage of maturity, with the presence of a mixture of green and brown grains, whereas the standard has almost no green grains as shown in Figure 10.

[0161] After threshing under the same conditions as for the previous tests on June 17 and 23, 2020, 3.5 kg of Isowheat immature grains and 2.8 kg of standard are available. See figure 10.

The immature grains harvested on July ¹ are subjected to steam cooking for 25 minutes using an SEB mini-compact kitchen steam cooker. Isowheat grains are considered sweeter, fresher, more tender, crunchier and less mealy than standard Apache immature grains, although the latter are appreciated. See figure 11. As in the other stages, most of the grains are frozen and another part is left to dry at 45° C. for 24 hours.

[0164] In tasting, the Isowheat native immature grains are judged for each harvest on June 17, 23 and July ¹ , 2021 to be sweeter with a fresher taste and a crunchier texture than the standard Apache native grain. It should be noted that the samples of Isowheat native grains have sweet flavors of different intensities depending on the harvest date, the most intense being obtained for the samples harvested on June 17, then on July ¹ and the least intense on June 23.

According to the years, on the basis of the climatic conditions, it will be necessary to adapt the date of harvest to obtain the optimal organoleptic qualities of the grains, which knows how to gauge the man of the art, wheat producer according to the characteristics of the invention.

[0165] Example 3 APPLICATIONS [0166] IMMATURE GRAINS [0167] The wheat grains according to the invention, hereinafter referred to as sweet wheat or Isowheat, used in this example were obtained by the method described above.

The standard reference wheats come from the APACHE variety, a commercial variety of soft winter wheat suitable for bread-making, produced and marketed by LIMAGRAIN LG Semences.

The Isowheat and Apache grains are planted in test plots with the cultivation route practiced by the person skilled in the art (the breeder) in Limagne in a conventional breeding program. Two stages of maturity were tested with grains harvested on June 23, 2020 and July ¹ , 2020. The earliest stage, grains harvested on June 23 (stage 1: humidity H for Isowheat of 69.2% and for Apache of 52 %) shows green grains for both varieties (attached figure 8) before cooking whereas for the other stage, grains harvested on July ¹ (stage 2: humidity H for Isowheat of 61.8% and for Apache of 41, 9% ) present for the two varieties a mixture of green and brown grains (figure 12 appended). It should be noted that the examination of the plants in plots as well as of the grains suggest a less advanced physiological stage on both dates for Isowheat compared to the Apache standard, as reflected for the Isowheat grains by a higher humidity and a greener color (stage 1) or with fewer brown grains (stage 2), as shown in Figure 12 attached. Isowheat native grains appear sweeter when tasted than the standard Apache wheat harvested on the same dates. Comparing Isowheat beans at both stages, they appear sweeter for stage 1 compared to stage 2.

[0169] Application: Water/Steam Cooking [0170] The grains are steam cooked using a SEB mini-compact kitchen steam cooker.

[0171] At stage 1 (humidity H for Isowheat of 69.2% and for Apache of 52%), after steaming for 20 minutes, the Isowheat sweet wheat grains are judged to be sweeter, more crunchy and better tasting. than standard Apache grains. See attached figure 9.

[0172] At stage 2 (humidity H for Isowheat of 61.8% and for Apache of 41.9%, immature Isowheat grains subjected to steam cooking for 25 minutes are judged to be sweeter, fresher, more tender, more crunchy and less mealy than standard immature grains even if the latter are appreciated (see figure 11 appended).

[01731 Application: Canned sweet corn type

[0174] Cans were made for the immature Isowheat and Apache grains harvested on the dates of June 17, 2020, June 23, 2020 and July ¹ , 2020. The grains are put in glass jars until they are three-quarters full. Brine, consisting of 1 teaspoon of salt for 1 liter of water, is then added to the top of the jars. The jars are hermetically sealed and sterilized at 115° C. for 30 minutes (see attached figure 13).

The following observations were made during the tasting:

Grains harvested on June 17, 2020: the product obtained with immature Isowheat grains is a little green, with an unpalatable appearance, the grain is well softened. The product has a correct texture in the mouth and can be consumed as is. The Apache control beans harvested on the same date are much drier, less pleasant and less sweet.

Grains harvested on June 23, 2020: the product obtained with immature Isowheat grains is much more palatable than that obtained with grains harvested on June 17, 2020, and has a more golden appearance with a good consistency in the mouth. They are juicier and considered very pleasant to taste. Apache control beans harvested on the same date are softer, less pleasant and less sweet. Grains harvested on July ¹ , 2020: the product obtained with immature Isowheat grains has firmer grains that stay in the mouth for a long time, but with a less interesting texture and taste than the samples harvested on June 23, 2020. The Apache control grains harvested by same date have a less pleasant texture in the mouth, more floury with a lot of chewiness.

The hydration of the Apache grains is more difficult than for the Isowheat grains, harvested on the same date, and are therefore all stuck together.

The tests indicate that whatever the harvest date, the canned Isowheat grains are judged to be more pleasant to taste, with a crispier texture and a sweeter taste than the control Apache grains. Preserves made with Isowheat grains harvested on June 23 are considered better than those made with Isowheat grains harvested on June 17 and July ¹ , 2020.

As indicated above, those skilled in the art will be able to adapt the harvest date to obtain the optimal organoleptic qualities. [0177] Application: Drying then poppaqe

The immature grains are dried in a conventional laboratory oven, for example of the Memmert® brand, at 45° C. for 24 hours. Drying gives them a crumpled appearance similar to that of mature grains. These dried immature grains can be treated like dry grains. Popping, carried out with a LISTO L3 kitchen popcorn machine (power of 1200W), restores the wrinkled Isowheat grain to a standard wheat grain shape and gives a product in all respects similar to that of mature grains popped with similar applications ( see attached figure 14).

[0179] Isowheat processed wheat harvested at an immature stage is suitable for applications tested on mature grains, after having undergone a stage of drying the immature grains to a moisture content equal to or less than 20% (see below). This allows the recovery of immature fruits deemed unsuitable for a specific immature grain application such as preserves for example. Another advantage is to also allow a delayed harvest of Isowheat grains, compared to that of a classic mature wheat allowing an advance treatment which is advantageous.

[0180] Application: Fried Grains

The immature Isowheat grains dried in a conventional laboratory oven, for example of the Memmert® brand, at 45° C. for 24 hours can be immersed in an oil at a temperature of 180° C. for 15 seconds. This operation will give them a puffy appearance and a crispy texture. [0182] MATURE GRAINS

[0183] Without treatment

[0184] Inclusion in laboratory bars. Prepare lab bread dough as indicated below: weigh 50g of flour, 32g of water, 1g of salt, 1g of yeast. Put the water first in the Kitchenaid® 4.3 liter 5K45SSEWH type mixer, then add the flour, salt and yeast. Switch on the kneader at speed 3 for 1 minute then switch to speed 5 for 5 minutes. Take the dough out, incorporate the Isowheat wheat grains into the dough, then ball it on the bench to form a very smooth ball before putting it in a kitchen paper mold (WORDIA white paper kitchen case, diameter 54mm, height 40mm). Put the dough to rise in the oven at 37°C for 40 minutes. Put the dough in the oven at 200°C for 15 minutes. After baking, let the lab bread cool for 1 hour at room temperature.

The Isowheat wheat grains incorporated into lab bread doughs swell during cooking and develop a crispy structure. [0186] Applications: o Topping on bread or pastry o Incorporation into bread-making or pastry products as a marking agent or providing crispness.

[0187] Application: Cooking with water or steam [0188] Steam cooking, for example by means of a SEB mini-compact kitchen steam cooker, causes the Isowheat grains to swell, which acquire a shape similar to that of a grain of standard wheat (see figures 9 and 11). The grains thus cooked have a crunchy texture with a fresh and sweet taste. They are popular with tasters, with some comparing them to Freekeh used in Middle Eastern cuisine. [0189] Applications: o Preserves o Cooked meals o Ebly® type (with mature grains) [0190] Application: Poppaqe LISTO L3® kitchen popcorn machine (power of 1200 W), the temperature measured in the machine during the test is 240° C., the duration of the popping varying from 35 to 45 seconds.

[0192] The popped Isowheat grains swell and change from a wrinkled shape to a standard wheat grain shape (see appended figure 14). They are crispy and sweet. Standard beans subjected to the same process are harder and have an overly starchy taste. Reviews from tasters are very positive. The duration of the process makes it possible to modulate the desired toasted perception according to the applications.

[0193] Possible uses: - Nutritional/health product

Snack: as is or salty or sweet coated.

- Topping: to add crispiness.

Breakfast cereals: incorporated into muesli for example Cereal bars

[01941 Application: Frying

The Isowheat grains are immersed in an oil at a temperature of 180° C. for 15 seconds. The same behavior is observed as during popping, the Isowheat grains swell, take the form of standard wheat grains and acquire a crispiness combined with a sweet taste (see appended figure 15). In terms of taste, being fried gives them great palatability. Same applications as for popped grains.

[0196] Application: Puffinq

The Isowheat and Apache wheat grains are sent to an expansion chamber at a temperature of 95°C, which is closed and pressurized to a given value by injecting steam. After a determined residence time, the chamber is opened, releasing the grains which, under the effect of the depression, will expand (puffer) (See diagram in appended figure 16). Depending on the pressure and residence time chosen, the grains will be more or less expanded.

Different tests are carried out with different pressure rises and residence times (see appended FIG. 17).

A: Native Apache grains, B: Apache grains puffed at 10 bars and for 10 seconds, C: Apache grains puffed at 21 bars and for 25 seconds, D: Native Isowheat grains, E: Isowheat grains puffed at 10 bars and for 10 seconds , F: Puffed Isowheat grains at 21 bars and for 25 seconds, G: Isowheat grains puffed at 12 bars and for 12 seconds.

The first test is carried out at a pressure of 10 bars for the Isowheat grains and for the Apache grains with a residence time of 10 seconds in the expansion chamber (see appended figure 17). The two types of Isowheat and Apache grains have a similar expanded appearance after puffing, which under the test conditions preserves the integrity of the grain. The expansion is much more marked for the Isowheat grains due to a native grain that is less filled (wrinkled) than the standard Apache grains. The second test is carried out at a pressure of 21 bars for the Isowheat grains and for the Apache grains with a residence time of 25 seconds in the expansion chamber (see appended figure 17). Under these conditions the expansion of the Apache grains results in a destructuring of the grains, the pericarp is no longer visible since under the effect of the pressure it has exploded to reveal the starchy albumen. Conversely, under the same conditions, the Isowheat grains did not expand, the envelope of the pericarp is still intact but under the effect of the residence time of 25 seconds at 95°C the grains burned. This lack of expansion is a reflection of the lack of starch in Isowheat wheat kernels. Another test with Isowheat grains is carried out at a pressure of 12 bars for 12 seconds at 95° C. and shows expanded grains with an intact envelope, without being burned (see appended figure 17).

[0201] Example 4 TEST PREFERENCE TEST ISOWHEAT BREAD vs COMPLETE BREAD CONTROL -30% SUGAR

[0202] ISOWHEAT

[0203] Preference test: The preference test is organized with a panel of tasters recruited on the site. It consists of the presentation of two products for which 4 questions are asked:

Question n°1: Take the sample between your hands, which sample do you prefer for its texture?

Question n°2: Put the sample in your mouth, which sample do you prefer for its texture?

Question n°3: Put the sample in your mouth, which sample do you prefer for its taste?

Question n°4: Put the sample in your mouth, which sample do you prefer for its aftertaste? For each question, tasters are asked to comment. Responses and comments are analyzed to establish the preferences expressed and their reasons.

[0204] A first evaluation was made between an Isowheat sandwich bread, that is to say prepared with integration in the recipe of 13% Isowheat integral flour and 30% less added sugar, compared to a control bread of bread full.

[0205] The test of preference between Isowheat sandwich bread and wholemeal control bread brought together 31 participants:

[0206] In terms of texture, Isowheat bread is preferred with 19 positive comments, 10 finding it softer, two comments indicate a bread that is too soft, which associated with softness is rather positive because the current consumer trend is for bread softer targeting teenagers and children.

[0207] In terms of texture in the mouth, 23 tasters prefer it, i.e. 74%; the control wholemeal bread is rated as rough and dry in comparison. [0208] In terms of taste, 19 tasters prefer it, i.e. 60%. Participants accustomed to consuming wholemeal bread preferred the control because Isowheat bread was, for them, not distinctive enough, without, however, making any negative comments about Isowheat bread. On the other hand, for the other tasters, the Isowheat bread is preferred to the complete control bread. No negative comments on the Isowheat bread were made by the panel of tasters, on the other hand, 5 negative comments were made on the control wholemeal bread. The Isowheat sandwich bread with 30% less sugar compared to the control wholemeal bread is considered more neutral, more pleasant, less rough than the control and 2 tasters find it even sweeter.

[0209] In terms of aftertaste, Isowheat sandwich bread is 75% preferred. Isowheat sandwich bread stays longer in the mouth with a sweeter, softer impression. The control is mentioned as having a bitter, sour, hay-like aftertaste that Iso does not leave behind.

[0210] At the level of overall preference: 70% of the participants prefer Isowheat sandwich bread with 30% less sugar than wholemeal control bread.

[0211] A second evaluation was made between Isowheat sandwich bread, that is to say prepared with integration in the recipe of 13% Isowheat wholemeal flour and 30% less added sugar compared to a control bread of classic white crumb recipe. The comparison test between Isowheat sliced bread and the white sliced yaws control brought together 33 participants. [0212] Visually, due to the integration of the whole flour, the Isowheat sandwich bread appears with a slightly gray crumb compared to the white crumb of the control sandwich bread.

[0213] In terms of texture to the touch, Isowheat sandwich bread is estimated to have the same level of preference as white sandwich bread, it is judged to be softer by 6 tasters and softer to the touch by 4 tasters.

[0214] In terms of texture in the mouth, the tasters did not note any notable differences, the Isowheat sandwich bread appears softer, for 4 tasters, than the control sandwich bread. It is important to note that Isowheat sliced bread is not found to be drier even though it is whole grain sliced bread compared to white sliced bread.

[0215] In terms of taste and aftertaste, the tasters issued very positive assessments for the Isowheat sandwich bread, 19 prefer it against 12 for the control sandwich bread. Isowheat sandwich bread is found to be sweeter, less bitter, softer than the control white sandwich bread. [0216] In terms of aftertaste, the Isowheat sandwich bread is considered to be more neutral, the sweet sensation persists longer despite 30% less added sugar compared to the control white sandwich bread.

[0217] In terms of overall preference, Isowheat sandwich bread recovers its initial visual impairment with 57% of tasters preferring it to control white sandwich bread.

[0218] Example 5 SUCROSE CONTENT IN MATURE ISOWHEAT AND

IMMATURE

[0219] Nutritional analyses, presented in Table 3 below, were carried out on mature grains of Isowheat wheat grains and Apache reference wheat grains, having the same genetic background, harvested over several years: 2017, 2019 & 2020. [0220] The fiber and protein contents are higher, approximately double for fiber and between 35 and 75% for protein in Isowheat wheat grains compared to Apache reference wheat. This can be explained by a deficiency in the biosynthesis of starch increasing de facto their contents.

These results show, in particular, a particularly high content of sucrose TS in the mature Isowheat grains. Indeed, depending on the harvest year, TS varies between 5.9% and 10.1%, i.e. 7 to 12 times more than the respective controls (always less than 1%). [0222] [TABLE 3]: Apache and Isowheat standard wheat nutritional analyzes over 3 harvest years

[0223] Sucrose content measurements were made on immature Isowheat grains sampled on three different dates, June 17 and 23 and July 1, 2020 and Apache harvested on June 17. The water contents of the Isowheat samples vary according to the harvest dates between 61% to 70%. The Apache control has a water content of 56%.

The differences in sucrose content expressed in matter as such MTQ (fresh matter) are between 4 to 6 times higher compared to the immature Apache control and between 6 and 9 times higher when expressed in dry matter.

[0225] [TABLE 41: sucrose content in immature Apache and Isowheat wheat grains

[0226] DM: dry matter [0227] MTQ: matter as such (fresh matter) [0228] Example 6 ISOWHEAT GRAIN GRINDING YIELD EVALUATION

[0229] Milling yield measurements are carried out according to standard NF EN ISO 27971 on three samples of Isowheat grains, two samples from two different lines harvested in 2019, Isowheat ISO 17-001 R19 and Isowheat ISO 17-002 R19, and one line harvested in 2020, Isowheat ISO 17-001 R20 PRUNS G2. Milling yields are compared to a reference made up of Apache grains with the same genetic background as the Isowheat lines except for the sweet wheat character, harvested in 2020.

These analyzes are carried out on a Chopin Dubois mill after wetting the wheat grains at a humidity of 16%. The wetting of wheat grain is a grain preparation operation, carried out before milling, with the aim of better separating the pericarp and the endosperm. This operation consists of bringing the grain to a moisture content of 16% by weight, to make the pericarp more flexible and the endosperm less brittle. Wetting consists, from a given batch of grain, in measuring its humidity to determine by calculation the quantity of water to add, to bring the humidity rate to 16% in relation to the total mass of the wheat grains. . To implement this wetting step, it is first necessary to determine the moisture content of the wheat grains used. The method used to determine the moisture content of wheat grains is the near infrared INFRATEC 1241 FOSS method, a predictive model (calibration) provided by the manufacturer FOSS. Knowing the mass of wheat grains to be wetted and their moisture content, the mass of water necessary for hydration is calculated. Advantageously, the wetting is carried out using devices known per se allowing the homogeneous distribution of the water of hydration within the grains of wheat. These devices are, for example, rotating containers (e.g. “BI-MIX” or “HYGROS-TEC III” from GOLFETTO SANGATI) or wetting screws.

[0232] A quantity of 800 g of wheat grains is placed in a container and the calculated quantity of water, to bring the grain to 16% humidity, is measured then divided into two equal quantities and is added twice . After each addition of water, the container is hermetically closed and placed on a rotary mixer for 30 minutes allowing homogenization of the water/grain mixture (mixing). The grain thus wetted is left to stand at room temperature for at least 24 hours.

[0233] Milling with the Chopin Dubois mill and according to standard NF EN ISO 27971 comprises two successive stages, grinding of the grains and conversion of the semolina. The grinding of wheat grains is carried out using grooved cylinders, resulting in fractions of large bran, semolina and grinding flour. The semolina fractions obtained during the grinding are taken up for a new passage in the mill between smooth cylinders. This operation, the conversion, is carried out in order to reduce the semolina into finer particles, distributed in remoulding (mixture of fatty flour and very fine bran) and conversion flours, separated by sieving. The number of conversions is repeated twice if the mass of semolina from the first grinding is greater than 48% of the initial grain weight.

To establish the milling yield, a measurement of the weights of the grinding and conversion flour fractions is made. These flours are combined into a total flour fraction whose humidity is determined to reduce the total weight of flour in dry matter. The milling yield is calculated by establishing the ratio between the weight, in dry matter, of the total flour produced and the weight, in dry matter, of the initial grain. The different fractions of the grinds of Apache and Isowheat grains are presented in figures 18, 19 and 20. [0235] This calculation is made according to the equation:

[0236] Milling yield = [((100- flour moisture) x total flour weight) / ((100- grain moisture) x grain weight)] x 100;

The results are presented in Table 5 below:

[0238] [TABLE 51: Grinding characteristics of Apache and Isowheat beans

It can be observed that the quantities of grinding and converting flours obtained from the Isowheat grains are extremely low compared to the Apache control. Indeed, on the three Isowheat samples, the grinding flour fraction does not exceed 2.5 g compared to 174.5 g for the control, and the weight of the converting flour reaches a maximum of 22 g whereas for the flour controls the weight is 569.7g.

This results in a much lower grinding yield for the Isowheat grains, less than 3%, compared to the control for which the grinding yield is 67.64%, ie 23 times higher. This difference is explained by a virtual absence of albumen in Isowheat wheat grains, giving them a crumpled appearance, and by the same, a virtual absence of starch granules which constitute on average 70% of white flour. .

[0241] This low grain filling, explaining a very low milling yield, can also be expressed indirectly through the “thousand grain weight” or PMG determined by the method according to standard NF EN ISO 520 January 2011. A “thousand grain weight” thousand grains” is a parameter that describes the accumulation capacity of reserve substances. A lower PMG, such as that measured in the Isowheat samples compared to the Apache control, may signify a deficit of starch synthesis (table

6).

[0242] [TABLE 6]: Thousand Grain Weight (PMG) of Apache and Isowheat grains

[0243] Another way of expressing the low filling of the grain can be indicated by the specific weight (SP) or apparent density of the grains, expressed in kilograms per hectolitre, and measured according to the method given in standard NF EN ISO 7971 -3:2019. A lower PS can be an indication of a starch deficiency as in the Isowheat samples for which the PS is around 56 kg/hl compared to 82 kg/hl for the Apache control line (Table 7).

[0244] [TABLE 7]: Specific weight (SP) of Apache and Isowheat grains

[0245] The flour obtained from Isowheat grains has a dark color due to the presence of contaminants from the fine bran fraction as also shown by the high ash content, between 2.1 and 2.64% compared to control 0, 57% (see table 5 above and photos of figures 18 and 20).

[0246] Sequence List

[0247] Sequence of the iso-1 gene located on chromosome 7AS of wheat Triticum aestivum of example 1, in which the genetic variation G→A (ApC-5022) appears, on exon 11:

[0248] Sequence of the iso-1 gene located on chromosome 7BS of wheat Triticum aestivum of example 1, in which the genetic variation G→A (ApA-2164) appears, on exon 4, and the genetic variation G →A (ApA-1119), on exon 9:

[0249] Sequence of the iso-1 gene located on chromosome 7DS of wheat Triticum aestivum of example 1, in which the genetic variation G→A (ApA-0866) appears, on exon 11

Claims

[Claim 1] Food product comprising processed wheat grains characterized in that these wheat grains have a Meunier Yield RM produced according to standard NF EN ISO 27971, expressed in % by mass of dry matter relative to whole wheat grains, such as, in ascending order of preference:

- 0.5 ≤ RM ≤ 10;

- 1 ≤ MR ≤ 5;

- 1.5 ≤ RM ≤ 3; and, preferably, a wrinkle-like phenotypic appearance.

[Claim 2] Food product according to claim 1, characterized in that these wheat grains have at least one of the following characteristics:

(-2.1-) Fiber content (FI), expressed as % by mass of material such as relative to whole wheat grains, such as, in increasing order of preference:

-10≤TF≤45;

-15≤TF≤40;

- 20≤ TF≤ 30;

(-2.2-) Ash content (TC), expressed in % by mass of material such as relative to whole wheat grains, such as, in increasing order of preference:

- 1.6 ≤ CT ≤ 3;

- 1.8 ≤ CT ≤ 2.8;

- 2 ≤ CT ≤ 2.5;

(-2.

3-) Protein content (TP), expressed in % by mass of material such as relative to whole wheat grains, such as, in ascending order of preference:

- 13≤TP≤35;

- 15 ≤ TP ≤ 30;

- 18 ≤ TP ≤ 25;

(-2.

4-) Total Sugar Content (TST), expressed as % by mass of matter such as relative to whole wheat grains, such as, in ascending order of preference:

- 1.5 ≤ TST ≤ 40;

- 2 ≤ TST ≤ 30;

- 2.5 ≤ TST ≤ 15.

- 4 ≤ ST ≤ 30;

- 5 ≤ ST ≤ 20; - 6 ≤ TS ≤ 16;

- 7 ≤ TS ≤ 15;

- 8 ≤ TS ≤ 14.

(-2.6-) phenotypic appearance of the crumpled type. [Claim 3] Food product according to Claim 1 or 2, characterized in that the wheat grains have a deficiency in at least one enzymatic activity involved in the biosynthesis and/or the structuring of amylopectin, which thwarts the formation of granules of starch, this deficiency preferably resulting from:

- a deficiency, total or partial, in the Persian activity of at least one enzyme involved in the biosynthesis and/or the structuring of amylopectin, which interferes with the formation of starch granules, and/or

- a deficiency, total or partial, in the expression of at least one gene coding for this (these) enzyme(s). [Claim 4] Food product according to at least one of Claims 1 to 3, characterized in that the wheat grains have on at least one of their genomes, preferably on at least 2 of their genomes, and, more preferably still, on all their genomes, at least one mutation of the iso-1 gene corresponding to the ISO-1 protein/enzyme and/or of the sh-2 gene corresponding to the SH-2 protein/enzyme, this mutation or these mutations resulting in a deficiency, total or partial, of the expression of the iso-1 gene and/or a deficiency, total or partial, of the expression of the sh-2 gene corresponding to SH-2, respectively.

[Claim 5] Food product according to Claim 3 or 4, characterized in that the deficiency results from a genetic variation identified within a Tilling population by means of specific markers.

[Claim 6] Food product according to at least one of the preceding claims, characterized in that it comprises wheat grains which are immature and which have a humidity H°, expressed in % by mass, such that, in ascending order preferably: - 16≤H°≤80;

- 45 ≤ H° ≤ 75;

- 55 ≤ H° ≤ 70.

[Claim 7] Food product according to at least one of Claims 1 to 5, characterized in that it comprises grains of wheat which are mature and which have a humidity H° measured, expressed in % by mass, such that, in increasing order of preference:

- 2 ≤ H° ≤ 16;

- 5 ≤ H° ≤ 16;

- 8 ≤ H° ≤ 15 .

[Claim 8] Food product according to Claim 6, characterized in that the transformation of the immature wheat grains consists of at least one of the following treatments: canning, freezing, drying, steaming, heating to a temperature of between 150 and 300° C., preferably between 170 and 250° C. at a heating rate chosen to allow the splitting of the grains (popping).

[Claim 9] Food product according to Claim 7, characterized in that the transformation of mature wheat grains consists of at least one of the following treatments:

(a) heating to a temperature between 150 and 300°C, preferably between 170 and 250°C at a heating rate chosen to allow the bursting of the grains (popping).

(b) Puffing.

(c) Toasting.

(d) Frying at a temperature between 160 and 200°C, preferably around 180°C for 5 to 60s, preferably 10 to 30s. (e) Flaking.

(f) Soaking in water.

(g) Steaming.

(h) Boiling

(i) Extrusion.

[Claim 10] Food product according to claim 7, characterized in that the transformation of the mature wheat grains consists of grinding to obtain semolina or integral flour, and optional heat treatment of this semolina or this flour.

[Claim 11] Bread products obtained from flour or semolina according to claim 10.

[Claim 12] Pastry products obtained from flour or semolina according to claim 10.

[Claim 13] Leaven, in particular liquid leaven, obtained from flour or semolina according to claim 10.

[Claim 14] Process of transformation making it possible to obtain the food product according to claim 6 or 8, characterized in that it consists: -1- in implementing immature wheat grains according to claim 6:

-3- and, possibly, to condition them.

[Claim 15]

Transformation process making it possible to obtain the food product according to at least one of claims 1 to 5, 7,9,10, characterized in that it consists of:

-1- to implement wheat grains according to claim 7;

-2- subjecting these wheat grains to at least one of the following treatments: (a) heating to a temperature of between 150 and 300°C, preferably between 170 and

250°C at a heating rate chosen to allow grain expansion (popping).

(b) Puffing.

(c) Toasting.

(d) frying at a temperature of between 160 and 200° C., preferably of the order of 180° C. for 5 to 60 s, preferably 10 to 30 s.

(e) flaking.

(f) soaking in water.

(g) steaming.

(h) extruding. -3- possibly conditioning them;

-4- and possibly canning them.

[Claim 16]

Wheat characterized in that it has on at least one of its genomes, preferably on at least 2 of its genomes, and, more preferably still, on all their genomes, at least one mutation of the iso-1 gene coding for the protein /ISO-1 enzyme and/or the sh-2 gene encoding the SH-2 protein/enzyme, this mutation or these mutations leading to a deficiency, total or partial, of the expression of the iso-1 gene and/or deficiency, total or partial, of the expression of the sh-2 gene corresponding to SH-2, respectively.