# Patent

## Title
Cereal flour composition containing wheat flour from low-temperature gelatinized wheat and food product using the same

## Abstract

An object of the present invention is to provide a cereal flour raw material that can provide products having superb texture. A further object of the present invention is to provide a food product manufactured using such a cereal flour raw material. The present invention provides a cereal flour composition that contains wheat flour prepared from a type of wheat that does not express any two of the three wheat starch synthase II proteins, and also does not express the three wheat granule bound starch synthase proteins, and another type of cereal flour; and a food product manufactured using such a cereal flour composition.

## Background
TECHNICAL FIELD
The present invention relates to a cereal flour composition containing wheat flour prepared from a novel wheat, and to a food product manufactured using such a cereal flour composition.
BACKGROUND ART
Wheat flour prepared from wheat is used in a wide range of applications, and a variety of processed foods are provided thereby. The characteristics of wheat flour are determined by many factors, but they are most strongly affected by the properties of the gluten and starch stored in the grains. In particular, starch generally accounts for 60 to 70% of the content of wheat flour, and its pasting properties have a large effect on the processing characteristics, appearance, and texture of the processed food. Starch originating in cereal grains can also come from rice and corn. In such plants searches for mutant genes involved in starch synthesis have been conducted, and mutants that store starch with properties different from the wild type have been discovered. However, because the chromosomes in wheat are hexaploid, mutations in the genes do not easily appear as a phenotype, and therefore very few such mutants are known in wheat.
Starch consists of two types of components: amylose wherein glucose is connected in a straight chain by an α-1,4 linkage, and amylopectin wherein glucose is connected in a branching structure by an α-1,6 linkage. Starch in wheat is a mixture of the two components of amylose wherein glucose is connected in a straight chain by an α-1,4 linkage, and amylopectin wherein glucose is connected in a branching structure by an α-1,6 linkage. These components are synthesized by the actions of various enzymes, and are stored in plants in the form of granules in the endosperm of the grains. With the addition of water and heat the starch granules gradually swell, and at a certain predetermined temperature (peak gelatinization temperature) the crystal structure breaks down, and the starch forms an amorphous paste (pasting). Subsequently, the gelatinized starch gradually increases in viscosity as it cools, and finally it gels (retrogradation).
It is known that such pasting properties and the amylose to amylopectin ratio vary greatly depending on the type of plant from which the starch is obtained. The common type of starch found in wheat has an amylose content of about 30%, and low-amylose wheat lines with a content of about 20% are also known. Low-amylose wheat starch is considered to be superior to conventional wheat starch as flour for making udon and other types of noodles, and it has been widely cultivated on a commercial scale. Types of rice and corn that store glutinous mochi (waxy) starch, which has an extremely low amylose content, were already known, and glutinous mochi (waxy) wheat was first bred by Nakamura et al. (see Patent document 1). Known enzymes in wheat that synthesize amylose are granule bound starch synthase-AI, granule bound starch synthase-BI, and granule bound starch synthase-DI, and Nakamura et al. produced waxy wheat by selecting a line in which none of these enzymes were expressed. This waxy wheat has unique processing characteristics and texture in comparison with conventional wheat types, and it is also considered to have improved retrogradation resistance and the like (see Patent documents 2 to 8). However, there have been few examples of practical application thereof because of problems with the processing characteristics or other texture problems.
On the other hand, wheat lacking starch synthase II-A1, starch synthase II-B1, and starch synthase II-D1, which are enzymes involved in the synthesis of amylopectin, has been reported (see Non-patent documents 1 and 2). It is stated that the amylose content in this type of wheat is markedly higher than in the wild type, but this wheat also has not reached practical application because it is not suited for processing into bread and other products.
[Patent document 1] Japanese Patent Application Laid-open No. H6-125669
[Patent document 2] Japanese Patent Application Laid-open No. H9-191818
[Patent document 3] Japanese Patent Application Laid-open No. H9-191819
[Patent document 4] Japanese Patent Application Laid-open No. H9-191842
[Patent document 5] Japanese Patent Application Laid-open No. H10-66511
[Patent document 6] Japanese Patent Application Laid-open No. H10-66527
[Patent document 7] Japanese Patent Application Laid-open No. H10-66529
[Patent document 8] Japanese Patent Application Laid-open No. H10-66530
[Non-patent document 1] Yamamori et al., Theor. Appl. Genet., 101:21-29 (2000)
[Non-patent document 2] Shimbata et al., Theor App. Genet., 111(6):1072-9 (2005)
DISCLOSURE OF THE INVENTION
An object of the present invention is to provide a cereal flour raw material that can provide a food product with superb texture. A further object of the present invention is to provide a food product manufactured using such a cereal flour raw material.
The inventors conducted extensive and intensive research to solve the above problems, and consequently they discovered that an excellent melt-in-the-mouth sensation, in addition to the previous characteristics of waxy wheat, is imparted by using flour from a type of wheat that does not express any two of the three wheat starch synthase II proteins, and also does not express the three wheat granule bound starch synthase proteins, and another type of cereal flour together, thus completing the present invention.
Therefore, the present invention is a cereal flour composition characterized in that it contains wheat flour prepared from a type of wheat that does not express any two of the three wheat starch synthase II proteins, and also does not express the three wheat granule bound starch synthase proteins, and also another type of cereal flour. In this case, the term wheat flour refers to a powder obtained by processing wheat, and it refers to a flour obtained by pulverizing whole wheat grains to produce a powder, or a flour classified by a difference in particle size obtained by passing that powder through a sieve, or a flour obtained by processing wheat into powder through a conventional flour milling process, and then mixing flour powders at a desired ratio, etc.
In the aforementioned cereal flour composition of the present invention, the content of wheat flour prepared from a type of wheat specified above is generally 1 to 50 wt %, preferably 1 to 30 wt %, and more preferably 2 to 20 wt %.
Therefore, modes for carrying out the present invention include the aforementioned cereal flour composition containing 1 to 50 wt % of the wheat flour prepared from a type of wheat that does not express any two of the three wheat starch synthase II proteins, and also does not express the three wheat granule bound starch synthase proteins; and the aforementioned cereal flour composition containing 1 to 30 wt % of the wheat flour prepared from a type of wheat that does not express any two of the three wheat starch synthase II proteins, and also does not express the three wheat granule bound starch synthase proteins. Furthermore, modes for carrying out the present invention include the aforementioned cereal flour composition containing 2 to 20 wt % of the wheat flour prepared from a type of wheat that does not express any two of the three wheat starch synthase II proteins, and also does not express the three wheat granule bound starch synthase proteins.
An example of the wheat flour used in the present invention that is prepared from a type of wheat that does not express any two of the three wheat starch synthase II proteins, and also does not express the three wheat granule bound starch synthase proteins is wheat flour prepared from a type of wheat that does not express the wheat starch synthase II-A1 protein and the wheat starch synthase II-B1 protein among the three wheat starch synthase II proteins, and also does not express the three wheat granule bound starch synthase proteins.
Therefore, one mode for carrying out the present invention is a cereal flour composition containing a wheat flour prepared from a type of wheat that does not express the wheat starch synthase II-A1 protein and the wheat starch synthase II-B1 protein among the three wheat starch synthase II proteins, and also does not express the three wheat granule bound starch synthase proteins, and also another type of cereal flour.
A different mode for carrying out the present invention is one wherein the aforementioned other type of cereal flour can be selected from the group consisting of wheat flour from non-waxy wheat, rice flour, starch, buckwheat flour, barley flour, corn flour, and oat flour. Among these, the use of wheat flour from non-waxy wheat is preferred.
The cereal flour composition of the present invention can be used in the manufacture of a variety of food products.
Therefore, the present invention is also a process for producing a food product that includes the use of the aforementioned cereal flour composition. The present invention is intended for the process of producing a food product that includes admixing a wheat flour prepared from a type of wheat that does not express any two of the three wheat starch synthase II proteins, and also does not express the three wheat granule bound starch synthase proteins together with another type of cereal flour. For the mix ratio of (A) a wheat flour prepared from a type of wheat that does not express any two of the three wheat starch synthase II proteins, and also does not express the three wheat granule bound starch synthase proteins, and (B) another type of cereal flour, a suitable ratio by weight for A:B is a range of 1:99 to 50:50, preferably 1:99 to 30:70, and more preferably 2:98 to 20:80.
The present invention is also intended for a food product manufactured using the aforementioned cereal flour composition. Examples of the above food product include bakery goods, noodles, deep fried foods, foods cooked on a griddle, roux and sauces, and products comprising a paste of meat or fish.
By combining the wheat flour that is prepared from a type of wheat that does not express any two of the three wheat starch synthase II proteins, and also does not express the three wheat granule bound starch synthase proteins together with another type of cereal flour in accordance with the present invention, it is possible to impart a moist, chewy texture, and also a soft, melt-in-the-mouth sensation.
The aforementioned superb texture is retained after the food product is manufactured, and even after it is stored at room temperature, or in a refrigerator or freezer.



BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 shows the pasting properties of wheat flour prepared from wheat lacking expression of the wheat starch synthase II-A1 and wheat starch synthase II-B1 proteins, and also lacking expression of the wheat granule bound starch synthase-A1, wheat granule bound starch synthase-B1, and wheat granule bound starch synthase-D1 proteins (LTG-SD wheat), and wheat flours prepared from other types of wheat.



BEST MODE FOR CARRYING OUT THE INVENTION
The wheat flour used in the present invention is wheat flour prepared from a type of wheat that does not express any two of the three wheat starch synthase II proteins, and also does not express the three wheat granule bound starch synthase proteins, i.e., it is wheat flour obtained from wheat (hereinafter referred to as LTG wheat) lacking expression of two proteins from among the wheat starch synthase II-A1, wheat starch synthase II-B1, and wheat starch synthase II-D1 proteins, and also lacking expression of the wheat granule bound starch synthase-A1, wheat granule bound starch synthase-B1, and wheat granule bound starch synthase-D1 proteins.
The above wheat can be any wheat lacking expression of two proteins from among the wheat starch synthase II-A1, wheat starch synthase II-B1, and wheat starch synthase II-D1 proteins, and also lacking expression of the wheat granule bound starch synthase-A1, wheat granule bound starch synthase-B1, and wheat granule bound starch synthase-D1 proteins. Preferably, the above wheat is lacking expression of the wheat starch synthase II-A1 and wheat starch synthase II-B1 proteins, and also lacking expression of the wheat granule bound starch synthase-A1, wheat granule bound starch synthase-B1, and wheat granule bound starch synthase-D1 proteins (hereinafter referred to as LTG-SD wheat).
In this case, the term “lacking expression” means that due to a mutation occurring in the genomic DNA thereof, synthesis of mRNA originating in the gene and synthesis of the enzyme protein itself do not occur. The above term also includes cases wherein, even if the protein is synthesized, a substitution or insertion, deletion, or other mutation arises in the amino acid sequence of the protein due to a mutation in the genomic DNA thereof, and all or part of the enzymatic function inherent in the wheat granule bound starch synthase proteins and wheat starch synthase II proteins, i.e., “the transfer of the glucose residue from the ADP-glucose substrate to the non-reducing end of the amylose recipient” or “the transfer of the glucose residue from the ADP-glucose substrate to the non-reducing end of the amylopectin recipient” is lost.
The above wheat can be obtained using any publicly known method of production and selection. As one example, a method can be noted wherein a type of wheat lacking expression of any two of the above three wheat starch synthase II proteins, and also lacking expression of the three wheat granule bound starch synthase proteins is selected from grains obtained by self-fertilization of F1 and subsequent generations of a type of wheat that was obtained by crossing wheat lacking the wheat starch synthase II proteins in accordance with the method described in Japanese Patent Application Laid-open No. 2005-333832 and a type waxy wheat obtained by the method described in Japanese Patent Application Laid-open No. H6-125669 following the methods described in Japanese Patent Application Laid-open Nos. 2005-333832 and H6-125669.
The present invention does not limit the crossing and selection method, and wheat obtained by an irradiation treatment or a chemical mutagenesis treatment, etc., or wheat obtained by gene recombination can be used. Furthermore, individuals among wheat lines that were raised using the above wheat parent crossing wheat lines and that lack expression of the above proteins can also be used. The wheat is selected using a method for detecting a mutation in the DNA thereof, or a method for qualitative and quantitative determination of the mRNA thereof by a reverse transcription reaction followed by PCR, or a method for qualitative or quantitative determination of the proteins contained in the grains thereof by SDS-PAGE, etc.
Wheat can also be used that is obtained by selecting the aforementioned type of wheat from progeny obtained by crossing the wheat obtained above and the wheat of another useful variety.
For example, it is possible to verify whether or not these proteins are expressed by detecting the presence or absence of the genes encoding the above three wheat starch synthase II proteins and the three wheat granule bound starch synthase proteins, or a mutation therein. Any method known to persons skilled in the art can be used as the method of verifying the expression of these proteins provided it is a method that can detect a mutation in the gene sequence, and examples thereof include methods such as PCR and the like.
The type of PCR method is not particularly limited herein, and various publicly known modified methods can be used. One example involves the mixing of a primer pair and template (test specimen) DNA together with reagents such as Tris-HCl, KCl, MgCl2, various dNTP, and TaqDNA polymerase to make the PCR reaction solution. One cycle of DNA PCR consists of the three steps of heat denaturation, primer annealing, and a DNA synthesis reaction by DNA polymerase. Because each step requires a different reaction temperature and reaction time, suitable ranges are established depending on the base sequence and length of the DNA region to be amplified. A thermal cycler for such a procedure is commercially available. Detection sensitivity can be increased by an investigation of preferred PCR conditions such as the concentrations of TaqDNA polymerase and MgCl2, the number of reaction cycles, etc., or the use of nested PCR.
The PCR reaction product can be identified using an immune reaction or can be identified by any other method, but if electrophoresis is performed, and clear bands can be recognized in an electrophoresis image using positive controls and negative controls whenever necessary, it is possible to verify the presence of the item to be detected (a gene encoding a protein and wheat carrying a genetic mutation) in the test sample.
The method described in Nakamura et al., Genome 45:1150-1156 (2002) can be used as the method for detecting mutations in the genes encoding the wheat granule bound starch synthase-A1, wheat granule bound starch synthase-B1, and wheat granule bound starch synthase-D1 proteins.
Examples of methods of detecting a mutation in genes encoding the wheat starch synthase II-A1, wheat starch synthase II-B1, wheat starch synthase II-D1, wheat granule bound starch synthase-A1, wheat granule bound starch synthase-B1, and wheat granule bound starch synthase-D1 proteins other than PCR include methods for qualitative or quantitative determination of the mRNA thereof by LAMP, NASBA, LCR, SDA, RCR, TMA, or RT-PCR.
The method described by Yamamori et al., Theor. Appl. Genet., 101:21-29 (2000) is one example of a method for verifying that the aforementioned wheat starch synthase II-A1, wheat starch synthase II-B1, and wheat starch synthase II-D1 proteins are not expressed.
The method described in Japanese Patent Application Laid-open No. H6-125669 can be used as the method for verifying that the aforementioned wheat granule bound starch synthase-A1, wheat granule bound starch synthase-B1, and wheat granule bound starch synthase-D1 proteins are not expressed.
The aforementioned LTG wheat produced in this manner has a lower gelatinization onset temperature and gelatinization peak temperature, and a higher maximum viscosity than wheat flour or starch obtained from publicly known waxy wheat. In addition, the energy value necessary for the gelatinization thereof is lower. Furthermore, when pasting is carried out by adding water to starch purified from LTG wheat and heating, and then the starch paste obtained thereby is stored long-term at a low temperature (4° C.), there is less change in viscosity of the starch paste and the clarity thereof can be retained longer than in a starch paste obtained from publicly known waxy wheat. Moreover, the branching chains constituting the amylopectin are shorter than in previous types of waxy wheat.
Therefore, wheat flour obtained from LTG wheat can be cooked at a lower temperature than the wheat flour obtained from publicly known waxy wheat, and its resistance to retrogradation is greater. The retrogradation of the product can be reduced thereby, and a superb melt-in-the-mouth sensation can be imparted to the product.
As a result, an entirely different processing method can be used for wheat flour obtained from LTG wheat than for wheat flour obtained from wheat previously known as waxy wheat, and a product with considerably different properties, texture and the like can be manufactured therefrom.
Examples of the wheat flour obtained from the aforementioned wheat used in the present invention include flour obtained by pulverizing whole wheat grains to produce a powder, or flour classified by a difference in particle size obtained by passing the powder through a sieve, or flour obtained by processing wheat into powder through a conventional flour milling process, and then mixing powders at a desired ratio. Or the wheat flour of the present invention also includes the starch purified therefrom.
The wheat flour obtained from LTG wheat can be made into a cereal flour composition with other cereal flours, and this cereal flour composition can be used for the manufacture of various foods. Moreover, the wheat flour obtained from the LTG wheat can be used for the manufacture of various foods along with other cereal flours.
The cereal flour composition of the present invention generally contains 1 to 50 wt % of the wheat flour obtained from LTG wheat and preferably 1 to 30 wt %. An even more preferred range is 2 to 20 wt %.
In the present invention, examples of cereal flours to be used with the wheat flour prepared from the LTG wheat include wheat flour from non-waxy wheat, rice flour, starch, buckwheat flour, barley flour, corn flour, and oat flour and the like. Wheat flour from non-waxy wheat includes general, commercially available hard flour, semi-hard flour, soft flour, durum semolina, or whole grain flour and the like. Among the above, the use of wheat flour from non-waxy wheat and durum semolina is preferred.
When food is manufactured, a suitable mix ratio of wheat flour (A) obtained from LTG wheat and other cereal flours (B) by weight is an A:B range of 1:99 to 50:50, a range of 1:99 to 30:70 is more suitable, and a range of 2:98 to 20:80 is even more suitable.
The foods manufactured in accordance the present invention are not particularly limited herein, and examples thereof include bakery goods, noodles, deep fried foods, foods cooked on a griddle, roux and sauces, and products comprising a paste of meat or fish.
Bakery goods refer to items that are produced by causing dough prepared with a base of wheat flour, etc., to rise using yeast, etc., and that are normally produced and sold in a bakery. Examples include breads such as conventional white bread, French bread, rolls, and pastries; deep-fried bread such as yeast doughnuts; steamed buns; pizza such as pizza pies; cakes such as sponge cake; and baked snacks such as cookies, biscuits, and the like.
Noodles refer to items produced by mixing salt, water, etc., with a base of wheat flour, etc., to obtain a kneaded dough, and after cutting off and shape-cutting a sheet thereof produced by rolling the dough with a rolling apparatus, etc., and depending on the intended use, processing by boiling, steaming, etc. Examples include udon noodles, Chinese soba, pasta, somen noodles, hiyamugi (cold) noodles, gyoza (pot sticker) wraps, shumai (steamed dumpling) wraps, and the like.
Moreover, the wheat flour obtained from the LTG wheat and other cereal flour can be mixed and used as a coating and batter for deep fried foods.
Deep fried foods refer to items wherein salt, flavoring, leavening agent, egg yolk, water and the like are mixed together and sprinkled on base ingredients such as vegetables, meat, seafood, and the like, which is then deep fried in oil. Examples include various types of Japanese deep fried foods such as tempura, kara-age, kaki-age, furai, tatsuta-age, and the like.
Additionally, examples of foods cooked on a griddle include items such as ohban-yaki, tako-yaki, okonomi-yaki, dora-yaki and the like wherein wheat flour is mixed with thickener, leavening agent, egg yolk, water, and the main ingredients, etc., as needed, to make a liquid dough, which is then cooked on a griddle, etc.
Moreover, it is possible to mix the wheat flour obtained from the LTG wheat and another cereal flour and, add that mixture as a thickener to roux and sauces. Examples of types of roux and sauces include curry sauce roux, hayashi (hash) sauce roux, stew sauce roux, and white sauce. Moreover, wheat flour obtained from the LTG wheat and other cereal flour can be mixed together and used as the binder for products comprising a paste of meat or fish. Examples of such products comprising a paste of meat or fish include patties, and boiled fish paste sticks such as chikuwa and kamahoko.
In bakery goods, it is suitable to use the cereal flour composition containing wheat flour obtained from the LTG wheat in a range of 1 to 50 wt %, preferably a range of 1 to 30 wt %, and even more preferably 2 to 20 wt %. In conventional white bread, for example, a preferred texture (softness, moistness, chewiness, fluffiness, etc.) and superb melt-in-the-mouth sensation can be obtained by using a cereal flour composition containing wheat flour obtained from LTG wheat especially in the range of 1 to 30 wt %, and more preferably 2 to 20 wt %. In sponge cakes, the preferred texture (chewiness, etc.) and a superb melt-in-the-mouth sensation can be obtained by using a cereal flour composition containing wheat flour obtained from LTG wheat especially in the range of 1 to 30 wt %, and more preferably 2 to 20 wt %. In cookies, the preferred texture (light texture, etc.) and a superb melt-in-the-mouth sensation can be obtained by using a cereal flour composition containing wheat flour obtained from LTG wheat especially in the range of 1 to 50 wt %, preferably 1 to 30 wt %, and even more preferably 2 to 20 wt %.
In general, a cereal flour composition containing wheat flour obtained from LTG wheat in the range of 1 to 50 wt %, preferably 1 to 30 wt %, and even more preferably 2 to 20 wt % can be used in noodles. In the case of udon noodles, for example, a texture with a balance of softness and viscoelasticity can be obtained by using a cereal flour composition containing wheat flour obtained from LTG wheat in the range of 1 to 50 wt %, preferably 1 to 30 wt %, and even more preferably 2 to 20 wt %.
In general, in foods cooked on a griddle it is suitable to use a cereal flour composition containing wheat flour obtained from LTG wheat especially in the range of 1 to 30 wt %, and more preferably 2 to 20 wt %. In the case of ohban-yaki, for example, the preferred texture (chewiness, lightness, etc.) and a superb melt-in-the-mouth sensation can be obtained by using a cereal flour composition containing wheat flour obtained from LTG wheat especially in the range of 1 to 30 wt %, and more preferably 2 to 20 wt %. In the case of tako-yaki, the preferred texture (chewiness, softness, aroma, etc.) and a superb melt-in-the-mouth sensation, and a creamy inside as well, can be obtained by using a cereal flour composition containing wheat flour obtained from LTG wheat in the range of 1 to 40 wt %, preferably 1 to 30 wt %, and even more preferably 2 to 20 wt %
The processes for producing various foods can be performed following conventional methods for those types of foods. All kinds of foods can be manufactured by properly selecting processing conditions other than inclusion of the aforementioned cereal flour such as the types of auxiliary ingredients to be used, types of additives to be used, mix ratio of the raw materials, processing temperature, processing time, and the like, and by using the manufacturing methods and equipment that are normally used.
Bakery goods, for example, can be produced by mixing various auxiliary ingredients that are normally used in the manufacture of bakery goods such as yeast, a chemical expander such as baking soda, yeast food, salt, sugar, oil or fat, egg, powdered milk, water and the like into the cereal flour composition of the present invention and kneading the same to prepare the dough. This is followed by either baking and frying the dough without rising or by letting the dough rise through fermentation, etc., and then baking or frying.
In the case of noodles, for example, dough is prepared by adding salt, water, and other auxiliary ingredients to the cereal flour composition of the present invention and kneading. The dough is made into a sheet by rolling the dough with a roller or rod-shaped item, and then the sheet is cut into a suitable size and shape. In the case of noodles such as udon, Chinese soba and the like, the dough is boiled. Gyoza and shumai are manufactured by passing through the steps of wrapping the ingredients in the dough, and suitably frying, steaming, or boiling.
Deep fried foods are manufactured by the steps of suitably mixing auxiliary ingredients such as starch, salt, leavening agent and the like into the cereal flour composition of the present invention, coating the main ingredients such as vegetables, meat, seafood, and the like with the mixture, and then deep frying in oil at a high temperature. Foods cooked on a griddle are manufactured by mixing salt, sugar, starch, leavening agent, thickener, coloring, egg, the main ingredients, etc., into the cereal flour composition of the present invention to make a liquid dough, and then cooking the same on a heated griddle or hot plate. Roux and sauces are manufactured by mixing butter and margarine with the cereal flour composition of the present invention, frying, adding soup base or milk thereto, and then boiling the mixture down.
EXAMPLES
[Test Materials]
1. Preparation, Selection, and Milling of LTG-SD Wheat
The LTG-SD wheat used for the present invention was prepared as follows. First three wheat lines, i.e., the generally known wheat line Kanto 79 (a wheat line lacking the wheat starch synthase II-B1 protein) and the foreign varieties Turkey 116 (a wheat line lacking the wheat starch synthase II-D1 protein) and Chosen 57 (a wheat lines lacking the wheat starch synthase II-A1 protein), were crossed in succession. A wheat line lacking all wheat starch synthase II proteins (but expressing all wheat granule bound starch synthase proteins) was selected from the progeny thereof, and that was used as one parent line. On the other hand, the Mochi-otome line was crossed with the imported varieties, and a waxy wheat (not expressing any of the wheat granule bound starch synthase proteins, but expressing all wheat starch synthase II proteins) selected from the F5 generation thereof was used as the other parent line, and as a comparative example.
These two wheat lines were crossed as parent lines, and the F1 generation was obtained. This was self-fertilized, F2 and subsequent generations were obtained, and expression of the wheat starch synthase II proteins in these progeny was verified by PCR (see Japanese Patent Application Laid-open No. 2005-333832). Moreover, following the method shown in Japanese Patent Application Laid-open No. H6-125669, expression of the wheat granule bound starch synthase proteins was verified by two dimensional electrophoresis, and a line was selected that does not express the wheat starch synthase II-A1 and wheat starch synthase II-B1 proteins, and does not express any of the wheat granule bound starch synthase proteins. Cultivation and crossing of all the wheat followed conventional methods.
A sufficient amount of these selected grains were propagated and used for the raw material in the tests. Water was added so that the water content of the harvested wheat reached 14%, and after it was let stand overnight, it was milled in a experimental mill made by Buhler Inc. All the powdered flour obtained from outlets 1B, 2B, 3B, 1M, 2M, and 3M were mixed together to make a straight flour. The yield rate of this milling was 61%. The flour obtained thereby was used in the tests.
2. Commercially Available Wheat Flour
EagleTM hard flour (Nippon Flour Mills), HeartTM soft flour (Nippon Flour Mills), ClubTM soft flour (Nippon Flour Mills), and Sanuki KikuTM flour (Nippon Flour Mills)
3. Wheat Flour from Norin 61 First Class Wheat (N61)
The Norin 61 wheat (a type expressing all wheat granule bound starch synthase proteins and all wheat starch synthase II proteins, hereinafter referred to as N61 wheat) was produced in Gunma Prefecture in 2006.
This was milled using the same procedure noted above for the LTG-SD wheat. The yield was 63%.
4. Wheat Flour from Waxy Wheat
The above waxy wheat cultivated at the National Agricultural Research Center for Tohoku Region was milled using the same procedure described above for the LTG-SD wheat. The yield was 58%.
5. Waxy Cornstarch
This product is widely used as waxy starch.
[Measurement of Pasting Properties]
The pasting properties of the flour powders prepared as described above were measured using a rapid visco analyzer (RVA). An RVA-4 manufactured by NEWPORT SCIENTIFIC was used, and the measurement method followed the standard method stipulated by the American Association of Cereal Chemists (AACC Method 76-21). First 3.5 g dry weight of powder from the prepared wheat flour was weighed out, 25 mL of 1 mM silver nitrate was added, and after this was stirred ten times by up and down movement with a stirrer, it was set in the apparatus. The measurement conditions were follows: After the speed was raised once to 960 rpm at a sample temperature of 50° C., the speed was reduced to 160 rpm while maintaining the temperature at 50° C., and those conditions were continued for 1 minute. Then the temperature was raised to 95° C. at a rate of 3° C./min while maintaining the speed, and after 10 min at 95° C., the sample was cooled to 50° C. at a rate of 3° C./min. The change in viscosity (cP) over this time was measured, and a graph was prepared therefrom. FIG. 1 shows the graph. The gelatinization peak temperature of each sample is shown below.













Peak temperature (° C.)



















N61 wheat
95.1



Waxy wheat
71.3



LTG-SD wheat
64.2










From these results it can be seen that the pasting properties of the LTG-SD wheat clearly show a lower gelatinization temperature than conventional N61 wheat and waxy wheat. It was known that the gelatinization peak temperature is lower in wheat flour from previously known waxy wheat than in conventional types of wheat, but these results reveal that the gelatinization peak temperature in the wheat flour from LTG-SD wheat is even lower. Moreover, the viscosity at the gelatinization peak temperature was also considerably higher. If the gelatinization peak temperature is lower, processing at a lower temperature becomes possible, and products with a very different texture and the like can be manufactured by selecting a processing method in accordance with those properties. For example, the effect of heating on the other ingredients can be reduced, or foods with a more advanced degree of gelatinization than previous products can be prepared.
[Measurement of Changes in Starch Paste During Storage Period]
The starch from the wheat flour obtained from the aforementioned waxy wheat and the LTG-SD wheat was purified, the starch was pasted, and retrogradation resistance was investigated.
Starch was purified from the straight flour powder obtained by milling each sample in the aforementioned Buhler experimental mill. To 100 g of the straight flour powder was added 48 mL of distilled water, and dough was kneaded at room temperature. This dough was rolled flat, after it had been divided into pieces of a suitable size, it was immersed in distilled water for one hour. The dough was folded together in the distilled water and gently squeezed until most of the starch was dispersed in the water. The suspension containing the starch was placed in a 500 mL centrifuge tube, and it was centrifuged for 10 min at 2500 rpm in a centrifuge (CS210 rotor No. 17N) manufactured by TOMY SEIKO CO., LTD. The supernatant was discarded, the remaining starch suspension was added, the procedure was carried out again in the same manner, and the precipitate was collected. Impurities such as pentosan that precipitated in the upper layer of the obtained precipitate were removed with a spatula, distilled water was added again and starch peret was suspended, and centrifugation was carried out. As many impurities as possible were removed by repeating this procedure, and the starch fraction was obtained. This starch fraction was once again suspended in a small amount distilled water, and after freezing at −80° C., it was lyophilized to obtain purified starch. After the water content of the purified starch was measured, a mathematically converted sample equivalent to 3.0 g with a water content of 14% was placed in a cup, distilled water was added to make a total weight of 28 g, and pasting of the starch was carried out by the RVA. For the pasting conditions, after the speed was raised once to 960 rpm at a sample temperature of 50° C., the speed was reduced to 160 rpm while maintaining the temperature at 50° C., and those conditions were continued for 1 minute. Then the temperature was raised to 95° C. at a rate of 3° C./min while maintaining the speed, and after 10 min at 95° C., the sample was cooled to 50° C. at a rate of 3° C./min. This starch paste was collected in a 50 mL plastic tube made by GREINER BIO-ONE, and stored sealed at 4° C. The viscosity of a gelatinized sample immediately after pasting and a gelatinized sample that had been stored for 4 days at a low temperature (4° C.) was measured using a B-type viscometer (TOKIMEC VISCOMETER model B8L). Measurement was carried out at a speed of 30 rpm using a No. 4 rotor. After being maintained at 25° C. for 1 hour, the samples were set in the instrument so that the rotor would be submerged in the sample up to 3.5 cm from the tip, the rotor was turned on, and the viscosity was measured after 30 sec. The results are shown below.














Immediately after pasting
Day 4




















Waxy wheat
9,700 mPa · s
10,780 mPa · s



LTG-SD wheat
8,040 mPa · s
 8,160 mPa · s










From the above results, it is clear that after pasting the increase in viscosity of the paste is lower in starch from the LTG-SD wheat than in starch from waxy wheat even after long-term storage at a low temperature.
Moreover, when the cloudiness of samples pasted in the same manner and stored at a low temperature (4° C.) was observed with the naked eye over time, the paste starch from LTG-SD wheat clearly was more transparent starting immediately after pasting. Additionally, in the starch paste from waxy wheat the extent of cloudiness increased with the passage of time in low temperature storage, but the starch paste from LTG-SD wheat maintained a high level of transparency. In samples stored 120 days after pasting, the starch paste from waxy wheat had become completely cloudy, and it had very little transparency, but the starch paste from the LTG-SD wheat retained a high level of transparency. In general, it can be said that starch becomes cloudy with retrogradation, and from the above findings it is clear that LTG-SD wheat is very resistant to retrogradation.
Additionally, using samples pasted in the same manner, the transparency immediately after pasting and after storage for 5 days at a low temperature (4° C.) was investigated.
An adequate amount of each sample was placed in a 1.5 mL tube, water was added at 100 times the amount of the sample, a good suspension was formed, and optical absorption at 372 nm was measured using a cell with a 1 cm optical path. The optical absorbance immediately after pasting was subtracted from the absorbance on day 5 to determine the increase in absorbance during low-temperature storage. The results are shown below.













Difference in absorbance



















Waxy wheat
0.099



LTG-SD wheat
0.050










From these results it is clear that the increase in optical absorbance during the low-temperature storage period was suppressed in the starch paste from LTG-SD wheat, and the starch paste retained its transparency.
The above results show that the starch paste from the LTG-SD wheat clearly has greater retrogradation resistance than starch from the waxy wheat. The extent of retrogradation resistance has a considerable effect on the texture of food products. Moreover, the deterioration of texture in food products can be prevented, and even greater long-term storage is enabled thereby.
Example 1
Manufacture of Conventional White Bread
Conventional white bread was manufactured using the following recipe (parts by weight).
















Wheat flour
100



Yeast food
0.1



Sugar
5



Salt
2



Skimmed milk
2



Shortening
5



Yeast
2



Water
68










The wheat flour in the above recipe was formulated using commercial wheat flour, wheat flour from LTG-SD wheat prepared as described above, wheat flour from N61 wheat, wheat flour from waxy wheat, and waxy cornstarch in the compositions (weight percent) shown in Table 1 below.
In this test EagleTM hard flour (Nippon Flour Mills) was used as the commercial flour. In the formulation table, the ingredients other than shortening were mixed in a mixer (SK MIXER CO., LTD., Type SK21C) for 2 min at low speed, 3 min at medium speed, and 1 min at high speed (27° C.). After the mixer was stopped and shortening was added, mixing was performed again for 1 min at low speed, 3 min at medium speed, and 5 min at high speed, and the kneaded dough was let rise for 60 min at 27° C. and 75% humidity. After the dough was punched down, rising was continued for 30 min under the same conditions, the dough was divided into 230 g portions, rolled into balls, and let stand for a bench time of 25 min. After the dough was reshaped in a molder, it was put into a shaped pan for final proofing until the dough had enlarged to about 80% of the pan in a rising chamber at 38° C. and 85% humidity, and then baked (205° C., 35 min). The baked bread was cooled by letting it stand for 1 hour at room temperature, placed into a plastic bag, let stand for 16 hours at room temperature, and sliced into 25 mm thick slices. Then an organoleptic evaluation was carried out.
The flavor and texture of the bread produced under the above conditions was evaluated on a 5-step scale by a ten member panel based on the following parameters and evaluation criteria, and an average score was calculated for each item.
Aroma


    5. Fragrant and strong aroma
    4. Rather strong aroma
    3. Aroma can be sensed
    2. No aroma
    1. Unpleasant aroma

Melt-in-the-Mouth Sensation

    5. Very good melt-in-the-mouth sensation
    4. Good melt-in-the-mouth sensation
    3. Average melt-in-the-mouth sensation
    2. Somewhat poor melt-in-the-mouth sensation
    1. Poor melt-in-the-mouth sensation, remains in the mouth

Hardness

    5. Soft
    4. Rather soft
    3. Average
    2. Rather hard
    1. Hard

Texture

    5. Extremely chewy
    4. Very chewy
    3. Flaky, but rather chewy
    2. Rather flaky
    1. Flaky












TABLE 1









Wheat flour content in formula (wt %)
Evaluation parameter





















Waxy

Melt-in-the-





Commercial
LTG-SD
N61
Waxy
corn-

mouth


Example
wheat
wheat
wheat
wheat
starch
Aroma
sensation
Hardness
Texture



















1-1
100
—
—
—
—
2.4
3.2
3.1
3.0


1-2
99.5
0.5
—
—
—
2.8
3.0
3.0
2.8


1-3
99
1
—
—
—
3.1
3.4
3.2
3.3


1-4
98
2
—
—
—
3.2
3.5
3.2
3.4


1-5
95
5
—
—
—
3.2
3.7
3.6
3.8


1-6
90
10
—
—
—
3.0
4.1
3.8
4.3


1-7
80
20
—
—
—
3.5
4.4
4.0
4.5


1-8
70
30
—
—
—
3.7
4.7
4.2
4.8


1-9
60
40
—
—
—
3.8
3.7
4.5
5.0


 1-10
50
50
—
—
—
4.1
2.1
5.0
5.0


 1-11
0
100
—
—
—
4.1
1.4
5.0
5.0


 1-12
90
—
10
—
—
2.2
2.6
2.7
2.6


 1-13
90
—
—
10
—
2.1
2.4
3.6
3.5


 1-14
90
—
—
—
10
2.0
2.2
3.4
3.3









After a portion of the above sliced bread was stored under refrigeration at 4° C. for 24 hours, the hardness of the bread surface was measured using a texture analyzer (STABLE MICRO SYSTEMS, TA, XT, plus). The machine was set so that an aluminum plunger (diameter 36 mm) would approach near the center of the sliced bread, and the bread was compressed with the plunger at a speed of 100 mm/min at a compression of 10 mm (40%). The load on the plunger was measured at a compression of 6.25 mm (25%). The measurement was performed on each sample 3 times, and the average values are summarized in Table 2. A lower numerical value indicates greater softness.










TABLE 2









Wheat flour content in formula (wt %)
















LTG-


Waxy




Commercial
SD
N61
Waxy
corn-


Example
wheat
wheat
wheat
wheat
starch
Hardness
















1-1 
100
—
—
—
—
554


1-6 
90
10
—
—
—
374


1-12
90
—
10
—
—
628


1-13
90
—
—
10
—
486


1-14
90
—
—
—
10
658









Bread using wheat flour from LTG-SD wheat had a much softer texture than bread containing conventional wheat flour, or bread wherein waxy wheat flour and waxy cornstarch were added to conventional wheat flour. Despite the emphasis on a chewy sensation, the bread had an extremely good mixture of melt-in-the-mouth sensation and flakiness in comparison with bread made using waxy wheat and waxy cornstarch. In addition, this tendency is retained even after storage under refrigeration for 24 hours. In particular, a content of 1 wt % or greater of flour from LTG-SD wheat with respect to the total amount of wheat flour is preferred, and from the standpoint of a suitable amount of softness, a content up to 30 wt % is most suitable. Even more preferable, in a formulation of 2 wt % or more, which makes these characteristics even clearer, a content up to 20 wt % is most suitable from the standpoint of no sticky sensation in the mouth.
Example 2
Manufacture of Sponge Cake
Sponge cake was manufactured using the following recipe (parts by weight).
















Wheat flour
100



Granulated sugar
125



Whole egg
175










The wheat flour in the above recipe was formulated using commercial wheat flour, wheat flour from LTG-SD wheat prepared as described above, wheat flour from N61 wheat, wheat flour from waxy wheat, and waxy cornstarch in the compositions (weight percent) shown in Table 3 below.
In this test HeartTM soft flour (Nippon Flour Mills) was used as the commercial flour. The process is as follows. The amount of whole egg necessary for the test was beaten beforehand and kept warm at 25° C. To 175 g of whole egg was added 125 g of granulated sugar and the mixture was mixed at low speed for 1 min, at high speed for 10 min, and again at low speed for 1 min. Following the compositions in Table 3, flour that had been mixed and sifted beforehand was added, and mixing was carried out at low speed for 30 sec. After the flour clinging to the walls was scraped down into the mixing bowl, mixing was resumed at low speed for 30 sec. The dough (360 g) was poured into a mold and baked for 30 min with a top setting of 185° C. and a bottom setting of 180° C. After the baked sponge cake was removed from the mold and let stand at room temperature for 30 min to cool, it was let stand at room temperature, and an organoleptic evaluation was performed the next day. The organoleptic evaluation was carried out by ten panelists based on the following parameters and evaluation criteria, and an average score was calculated for each item. The same evaluation was carried out after the manufactured sponge cake had been stored in a refrigerator at 4° C. for 24 hours. Table 3 shows the results after storage at room temperature, and Table 4 shows the results after storage under refrigeration.
Aroma


    5. Fragrant and strong aroma
    4. Rather strong aroma
    3. Aroma can be sensed
    2. No aroma
    1. Unpleasant aroma

Melt-in-the-Mouth Sensation

    5. Very good melt-in-the-mouth sensation
    4. Good melt-in-the-mouth sensation
    3. Average melt-in-the-mouth sensation
    2. Somewhat poor melt-in-the-mouth sensation
    1. Poor melt-in-the-mouth sensation, remains in the mouth

Sweetness

    5. Extremely sweet
    4. Very sweet
    3. Rather sweet
    2. Slightly sweet
    1. Sweetness lacking

Texture

    5. Soft and very chewy
    4. Very chewy
    3. Rather chewy
    2. Slightly chewy
    1. Not chewy












TABLE 4









Wheat flour content in formula (wt %)
Evaluation parameter
















Examples




Waxy

Melt-in-the-




stored in
Commercial
LTG-SD
N61
Waxy
corn-

mouth


refrigerator
wheat
wheat
wheat
wheat
starch
Aroma
sensation
Sweetness
Texture



















2-1
100
—
—
—
—
2.5
2.1
2
2


2-2
99.5
0.5
—
—
—
2.2
2.3
2
2.1


2-3
99
1
—
—
—
2.1
2.3
1.9
2.3


2-4
98
2
—
—
—
2.3
2.5
2.2
2.5


2-5
95
5
—
—
—
2.3
2.6
2.1
2.7


2-6
90
10
—
—
—
2.4
2.6
2.3
3


2-7
80
20
—
—
—
2.6
2.8
2.4
3.1


2-8
70
30
—
—
—
2.4
2.9
2.3
3.2


2-9
60
40
—
—
—
2.7
3
2.6
3.2


 2-10
50
50
—
—
—
2.9
2.8
2.7
3.3


 2-11
0
100
—
—
—
3.2
1.5
3
2.4


 2-12
90
—
10
—
—
2.1
1.9
2.2
1.8


 2-13
90
—
—
10
—
2.2
1.5
2.3
2.5


 2-14
90
—
—
—
10
2
1.4
2.5
2.3


















TABLE 3







Examples
Wheat flour content in formula (wt %)
Evaluation parameter
















stored




Waxy

Melt-in-the-




at room
Commercial
LTG-SD
N61
Waxy
corn-

mouth


temp.
wheat
wheat
wheat
wheat
starch
Aroma
sensation
Sweetness
Texture



















2-1
100
—
—
—
—
3
3
2
2


2-2
99.5
0.5
—
—
—
3.1
3
1.9
2.2


2-3
99
1
—
—
—
3.1
3.2
2
2.3


2-4
98
2
—
—
—
3
3.4
2.3
2.8


2-5
95
5
—
—
—
3.4
3.4
2.5
3.2


2-6
90
10
—
—
—
3.5
3.6
2.8
3.7


2-7
80
20
—
—
—
3.3
3.7
2.8
3.9


2-8
70
30
—
—
—
3.5
3.7
3
4.2


2-9
60
40
—
—
—
3.3
3.6
3.1
4.5


 2-10
50
50
—
—
—
3.5
2.7
3.3
4.7


 2-11
0
100
—
—
—
4.1
1.5
4.2
3.1


 2-12
90
—
10
—
—
2.8
2.5
2.1
1.8


 2-13
90
—
—
10
—
3.6
2.1
2.5
3.6


 2-14
90
—
—
—
10
2.7
2
2.7
3.1









From these results it is clear that when wheat flour from the LTG-SD wheat, waxy wheat flour, and waxy cornstarch are used, the LGT-SD wheat imparts greater chewiness and softer texture than in previous products. When waxy wheat flour and waxy cornstarch are used, the cake forms lumps in the mouth, and the melt-in-the-mouth sensation is poor. However, when wheat flour from the LTG-SD wheat is used, the texture is soft and the melt-in-the-mouth sensation is good. This tendency is retained even after 24-hour storage under refrigeration.
Although the chewiness increases together with the content of wheat flour from LTG-SD wheat, the melt-in-the-mouth sensation becomes poorer, and the volume of the product also becomes smaller. From this viewpoint, a mix with a range of 1 to 40 wt % of wheat flour from LTG-SD wheat, and especially 1 to 30 wt % with respect to the total amount of wheat flour is preferred, and a mix with a range of 2 to 20 wt % is even more preferred.
Example 3
Manufacture of Cookies
Cookies were manufactured using the following recipe (parts by weight).
















Wheat flour
150



Granulated sugar
75



Nonfat powdered milk
5



Salt
1



Shortening
50



Baking soda
0.6



Ammonium bicarbonate
0.8



Water
30










The wheat flour in the above recipe was formulated using commercial wheat flour, wheat flour from LTG-SD wheat prepared as described above, wheat flour from N61 wheat, wheat flour from waxy wheat, and waxy cornstarch in the compositions (weight percent) shown in Table 5 below.
In this test HeartTM soft flour (Nippon Flour Mills) was used as the commercial flour. The process is as follows. The required amounts of granulated sugar, nonfat powdered milk, salt, shortening, baking soda, ammonium bicarbonate, and water were combined and mixed in a mixer at low speed for 1 min. Then mixing was carried out at high speed for 4 min, and after the flour clinging to the walls was scraped down into the mixing bowl, mixing was resumed for 4 min. After the weighed out amount of wheat flour was added and mixing was carried out at low speed for 30 seconds, the flour clinging to the walls was scraped down into the mixing bowl, and mixing was resumed at low speed for 30 seconds. The dough was removed from the mixer, folded together, and shaped into a rod. This rod-shaped dough was divided into six parts and each part was spread out on an aluminum plate to a thickness of 6 mm. Cookies were cut in round shapes 6 cm in diameter and baked for 12 minutes (top setting 200° C., bottom setting 210° C.). After the cookies were let stand to cool at room temperature for 15 min/side, they were placed in a plastic bag and stored at room temperature. An organoleptic evaluation was carried out the next day. Evaluation on a 5-step scale was carried out by a ten member panel based on the following parameters and evaluation criteria, and an average score was calculated for each item. The results are shown in Table 5.
Aroma


    5. Fragrant and strong aroma
    4. Rather strong aroma
    3. Aroma can be sensed
    2. No aroma
    1. Unpleasant aroma

Melt-in-the-Mouth Sensation

    5. Very good melt-in-the-mouth sensation
    4. Good melt-in-the-mouth sensation
    3. Average melt-in-the-mouth sensation
    2. Somewhat poor melt-in-the-mouth sensation
    1. Poor melt-in-the-mouth sensation, remains in the mouth

Flavor

    5. Considerably sweet in addition to normal flavor
    4. Sweetness slightly emphasized in addition to normal flavor
    3. Normal flavor
    2. Rather weak flavor
    1. Weak flavor

Consistency

    5. Considerably soft
    4. Slightly soft
    3. Right amount of hardness
    2. Slightly hard
    1. Hard and crumbly












TABLE 5









Wheat flour content in formula (wt %)
Evaluation parameter





















Waxy

Melt-in-





Commercial
LTG-SD
N61
Waxy
corn-

the-mouth


Examples
wheat
wheat
wheat
wheat
starch
Aroma
sensation
Flavor
Consistency



















3-1
100
—
—
—
—
3.2
3
3.1
3.1


3-2
99.5
0.5
—
—
—
3.2
3.1
3
3.1


3-3
99
1
—
—
—
3.2
3.2
3.1
3.2


3-4
98
2
—
—
—
3.4
3.5
3.5
3.4


3-5
95
5
—
—
—
3.6
3.7
3.6
3.7


3-6
90
10
—
—
—
3.5
4.1
3.7
3.9


3-7
80
20
—
—
—
3.8
4.1
3.8
4.2


3-8
70
30
—
—
—
3.9
4.3
3.7
4.4


3-9
60
40
—
—
—
3.9
4.4
4.1
4.6


 3-10
50
50
—
—
—
3.8
4
4.3
4.2


 3-11
40
60



3.9
3.8
4.3
4.3


 3-12
30
70



3.9
3.2
4.4
4.3


 3-13
0
100
—
—
—
3.7
2.8
4.3
4.5


 3-14
90
—
10
—
—
3
3.1
2.9
3.2


 3-15
90
—
—
10
—
2.9
2.7
3.2
3.6


 3-16
90
—
—
—
10
3.1
2.5
3.3
3.2









From these results it is clear that when wheat flour from the LTG-SD wheat is used, the product has a lighter texture, the inside is moister, and the melt-in-the-mouth sensation is better than when conventional waxy flour is used. However, when the content of flour from LTG-SD wheat with respect to the total amount of wheat flour became too large, the cookies had a conspicuous watery texture. From this viewpoint, a mix with a range of 1 to 50 wt %, and especially 1 to 30 wt % with respect to the total amount of wheat flour is preferred, and a mix with a range of 2 to 20 wt % is even more preferred.
Example 4
Manufacture of Ohban-Yaki (a Small Round Pancake Filled with Sweet Bean Paste)
Ohban-yaki were manufactured using the following recipe (parts by weight).
















Wheat flour
89



Sugar
3



Glucose
5



Baking powder
3



Water
120










The wheat flour in the above recipe was formulated using commercial wheat flour, wheat flour from LTG-SD wheat prepared as described above, wheat flour from N61 wheat, wheat flour from waxy wheat, and waxy cornstarch in the compositions (weight percent) shown in Table 6 below.
In this test ClubTM soft flour (Nippon Flour Mills) was used as the commercial flour. The process is as follows. The ingredients in the above recipe were mixed together and stirred with a whisk to make batter. The batter was poured onto the griddle of an ohban-yaki maker set to 180° C., the bean jam was placed over the batter, and after cooking for 5.5 min, the griddle was flipped over and cooking was continued for another 5.5 min. An organoleptic evaluation was carried out on the cooked ohban-yaki immediately after preparation and on ohban-yaki that were placed in a refrigerator for 24 hours after cooking and then warmed in a microwave oven. The organoleptic evaluation was carried out using a 5-step scale by a ten member panel based on the following parameters and evaluation criteria, and an average score was calculated for each item. Table 6 shows the results immediately after cooking, and Table 7 shows the results after storing under refrigeration.
Aroma


    5. Fragrant and strong aroma
    4. Rather strong aroma
    3. Aroma can be sensed
    2. No aroma
    1. Unpleasant aroma

Melt-in-the-Mouth Sensation

    5. Very good melt-in-the-mouth sensation
    4. Good melt-in-the-mouth sensation
    3. Average melt-in-the-mouth sensation
    2. Somewhat poor melt-in-the-mouth sensation
    1. Poor melt-in-the-mouth sensation, remains in the mouth

Crispiness (Feels Good to Bite)

    5. Good crispiness
    4. Rather good crispiness
    3. Average crispiness
    2. Rather lacking in crispiness and a little doughy.
    1. Lacking in crispness and doughy.

Chewiness

    5. Soft and very chewy
    4. Very chewy
    3. Rather chewy
    2. Not very chewy
    1. Not chewy at all











TABLE 6







Examples
Wheat flour content in formula (wt %)
Evaluation parameter
















immediately




Waxy

Melt-in-




after
Commercial
LTG-SD
N61
Waxy
corn-

the-mouth


cooking
wheat
wheat
wheat
wheat
starch
Aroma
sensation
Crispiness
Chewiness



















4-1
100
—
—
—
—
4.1
3.1
3.2
3.2


4-2
99.5
0.5
—
—
—
3.9
3.0
3.0
3.1


4-3
99
1
—
—
—
4.0
3.2
3.3
3.4


4-4
98
2
—
—
—
3.9
3.3
3.2
3.5


4-5
95
5
—
—
—
3.9
3.5
3.4
3.7


4-6
90
10
—
—
—
4.0
3.6
3.0
3.9


4-7
80
20
—
—
—
4.0
3.8
2.9
4.2


4-8
70
30
—
—
—
4.1
3.4
2.5
4.5


4-9
60
40
—
—
—
4.2
2.7
1.9
4.6


 4-10
50
50
—
—
—
3.8
1.2
1.4
5.0


 4-11
0
100
—
—
—
3.5
1.1
1.2
4.7


 4-12
90
—
10
—
—
3.8
2.5
2.8
2.7


 4-13
90
—
—
10
—
4.0
2.3
2.6
3.7


 4-14
90
—
—
—
10
3.8
2.4
2.5
3.3


















TABLE 7







Examples
Wheat flour content in formula (wt %)
Evaluation parameter
















after




Waxy

Melt-in-




storage in
Commercial
LTG-SD
N61
Waxy
corn-

the-mouth


refrigerator
wheat
wheat
wheat
wheat
starch
Aroma
sensation
Crispiness
Chewiness



















4-1
100
—
—
—
—
3.1
2.3
2.1
2.4


4-2
99.5
0.5
—
—
—
2.8
2.2
2.3
2.2


4-3
99
1
—
—
—
2.6
2.5
2.2
2.4


4-4
98
2
—
—
—
2.5
2.5
2.3
2.6


4-5
95
5
—
—
—
2.2
2.7
2.5
3.0


4-6
90
10
—
—
—
2.3
2.9
2.5
3.2


4-7
80
20
—
—
—
2.5
2.8
2.3
3.5


4-8
70
30
—
—
—
2.4
2.6
2.1
3.7


4-9
60
40
—
—
—
2.7
2.2
2.0
3.8


 4-10
50
50
—
—
—
2.5
1.8
1.7
2.5


 4-11
0
100
—
—
—
2.2
1.2
1.3
1.6


 4-12
90
—
10
—
—
2.4
2.0
2.1
1.9


 4-13
90
—
—
10
—
2.8
2.1
2.3
2.9


 4-14
90
—
—
—
10
2.5
2.3
2.4
2.1









From the above results it is clear that when flour from the LTG-SD wheat was included the product becomes very chewy and has a good melt-in-the-mouth sensation. In addition, this tendency is retained even after storage of the food under refrigeration. A content of 1 to 30 wt %, and more preferably a content of 2 to 20 wt %, with respect to the total wheat flour was considered to be very suitable.
Example 5
Manufacture of Tako-Yaki (a Dumpling Made of Batter, Diced Octopus, and Flavorings)
Tako-yaki were manufactured using the following recipe (parts by weight).
















Wheat flour
92



Gluocse
3



Granulated sugar
2



Salt
2



Powdered dried bonito
1



Water
210



Egg
5










The wheat flour in the above recipe was formulated using commercial wheat flour, wheat flour from LTG-SD wheat prepared as described above, wheat flour from N61 wheat, wheat flour from waxy wheat, and waxy cornstarch in the compositions (weight percent) shown in Table 8 below.
In this test ClubTM soft flour (Nippon Flour Mills) was used as the commercial flour. The process is as follows. The ingredients in the table were mixed together and stirred with a whisk to make batter. The batter was poured onto the griddle of a tako-yaki maker set to 180° C., and after cooking for 4 min, a hot shaping griddle heated to 180° C. was placed on top and cooking was continued for another 1 min. An organoleptic evaluation was carried out on the cooked tako-yaki immediately after preparation and on tako-yaki that were placed in a freezer at −20° C. for 24 hours after cooking and then warmed in a microwave oven. The organoleptic evaluation was carried out using a 5-step scale by a ten member panel based on the following parameters and evaluation criteria, and an average score was calculated for each item. Table 8 shows the results immediately after cooking, and Table 9 shows the results after frozen storage.
Aroma


    5. Fragrant and strong aroma
    4. Rather strong aroma
    3. Aroma can be sensed
    2. No aroma
    1. Unpleasant aroma

Melt-in-the-Mouth Sensation

    5. Very good melt-in-the-mouth sensation
    4. Good melt-in-the-mouth sensation
    3. Average melt-in-the-mouth sensation
    2. Somewhat poor melt-in-the-mouth sensation
    1. Poor melt-in-the-mouth sensation, remains in the mouth

Chewiness

    5. Soft and very chewy
    4. Very chewy
    3. Rather chewy
    2. Not very chewy
    1. Not chewy at all











TABLE 8







Examples
Wheat flour content in formula (wt %)
Evaluation parameter















immediately




Waxy

Melt-in-



after
Commercial
LTG-SD
N61
Waxy
corn-

the-mouth


cooking
wheat
wheat
wheat
wheat
starch
Aroma
sensation
Chewiness


















5-1
100
—
—
—
—
3.3
3.1
2.5


5-2
99.5
0.5
—
—
—
3.1
3.0
2.7


5-3
99
1
—
—
—
3.1
3.3
2.8


5-4
98
2
—
—
—
3.2
3.5
3.1


5-5
95
5
—
—
—
3.0
3.7
3.4


5-6
90
10
—
—
—
3.3
3.8
3.9


5-7
80
20
—
—
—
3.1
4.1
4.5


5-8
70
30
—
—
—
3.4
3.7
4.6


5-9
60
40
—
—
—
3.3
3.1
4.9


 5-10
50
50
—
—
—
2.9
2.7
4.6


 5-11
0
100
—
—
—
3.0
1.8
4.8


 5-12
90
—
10
—
—
3.1
2.9
2.8


 5-13
90
—
—
10
—
3.1
2.9
3.4


 5-14
90
—
—
—
10
2.8
2.7
3.1


















TABLE 9







Examples
Wheat flour content in formula (wt %)
Evaluation parameter















after




Waxy

Melt-in-the



frozen
Commercial
LTG-SD
N61
Waxy
corn-

mouth


storage
wheat
wheat
wheat
wheat
starch
Aroma
sensation
Chewiness


















5-1
100
—
—
—
—
2.8
2.4
2.2


5-2
99.5
0.5
—
—
—
2.4
2.3
2.4


5-3
99
1
—
—
—
2.5
2.3
2.3


5-4
98
2
—
—
—
2.1
2.4
2.6


5-5
95
5
—
—
—
2.4
3.1
2.8


5-6
90
10
—
—
—
2.5
3.0
3.1


5-7
80
20
—
—
—
2.7
2.8
3.2


5-8
70
30
—
—
—
2.6
2.5
3.3


5-9
60
40
—
—
—
2.8
2.1
3.6


 5-10
50
50
—
—
—
2.8
1.7
3.7


 5-11
0
100
—
—
—
3.0
1.5
3.6


 5-12
90
—
10
—
—
2.5
2.2
2.0


 5-13
90
—
—
10
—
2.6
2.2
2.3


 5-14
90
—
—
—
10
2.4
2.0
2.1









From the above results it is clear that when flour from LTG-SD wheat was included, the product becomes very chewy and has a good melt-in-the-mouth sensation. In addition, this tendency is retained even after frozen storage. However, when the content of flour from LTG-SD wheat with respect to the total amount of wheat flour became too large, shape retention tended to be poor because the voids formed inside the tako-yaki grew too large and then contracted too much after cooking, etc. From this viewpoint, a mix with a range of 1 to 40 wt %, and preferably 1 to 30 wt %, of wheat flour from LTG-SD wheat with respect to the total amount of wheat flour is suitable. Even more preferred is a mix with 2 to 20 wt % thereof.
Example 6
Manufacture of Udon Noodles
Udon noodles were manufactured using the following recipe (parts by weight).
















Wheat flour
500



Salt
10



Water
170










The wheat flour in the above recipe was formulated using commercial wheat flour, wheat flour from LTG-SD wheat prepared as described above, wheat flour from N61 wheat, and in the compositions (weight percent) shown in Table 10 below.
In this test Sanuki KikuTM flour (Nippon Flour Mills) was used as the commercial flour. The process is as follows. Water with the salt dissolved therein was added to the wheat flour in a mixer with stirring, and the mixture was kneaded for 5 min. The small crumbs of dough were shaped by passing them through a noodle roll. Compounding was carried out by folding the formed dough and passing it through the noodle roll again. Then the dough was spread until it reached a thickness of 2.5 mm. The sheet was then cut using a No. 10 square bladed roller and stored at 4° C. Water with an adjusted pH was boiled, and after the noodles were cooked in the boiling water for 21 min, the noodles were unraveled in cold water, placed on a bamboo basket, covered with plastic wrap, and let stand 30 min at room temperature. An organoleptic evaluation of the udon prepared in the above manner was carried out by a ten member panel based on the following parameters and evaluation criteria using example 6-1, which had a commercial flour content of 100 wt % as the flour component, as a reference standard. An average score was calculated for each item. Table 10 shows the results.
Softness


    5. Soft
    4. Rather soft
    3. Medium soft
    2. Rather hard
    1. Hard

Viscoelasticity

    5. Excellent visoelasticity
    4. Rather good visoelasticity
    3. Medium visoelasticity
    2. Rather poor visoelasticity
    1. Poor visoelasticity

Smoothness

    5. Excellent smoothness
    4. Rather good smoothness
    3. Medium smoothness
    2. Rather poor smoothness
    1. Poor smoothness












TABLE 10









Wheat flour content in formula (wt %)
Evaluation parameter















Commercial
LTG-SD
N61
Waxy

Visco-



Examples
wheat
wheat
wheat
wheat
Softness
elasticity
Smoothness

















6-1
100
—
—
—
3.0
3.0
3.0


6-2
99
1
—
—
3.1
3.1
3.1


6-3
98
2
—
—
3.1
3.3
3.1


6-4
95
5
—
—
3.3
3.7
3.2


6-5
90
10
—
—
3.4
3.8
3.4


6-6
80
20
—
—
3.7
3.9
3.3


6-7
70
30
—
—
3.8
4.1
3.5


6-8
60
40
—
—
4.0
4.0
3.2


6-9
50
50
—
—
4.2
3.8
3.1


 6-10
40
60
—
—
4.5
3.4
2.9


 6-11
0
100
—
—
4.7
2.1
2.5


 6-12
90
—
10
—
3.1
2.9
3.1


 6-13
90
—
—
10
3.2
3.6
3.0









From the above results it is clear that when flour from LTG-SD wheat was included, the product becomes chewy and has a good balance of viscoelasticity and softness. However, when the content became too great the noodles tended to become too soft. As a result, a mix with a range of 1 to 50 wt % of the wheat flour from LTG-SD wheat with respect to the total amount of wheat flour, and from the viewpoint of superior elasticity, preferably 1 to 30 wt %, and more preferably 2 to 20 wt %, is suitable.

## Claims

1. A cereal flour composition that contains wheat flour prepared from a type of wheat that does not express any two of the three wheat starch synthase II proteins, and also does not express the three wheat granule bound starch synthase proteins, and another type of cereal flour.

2. The cereal flour composition according to claim 1 that contains 1 to 50 wt % of the wheat flour prepared from a type of wheat that does not express any two of the three wheat starch synthase II proteins, and also does not express the three wheat granule bound starch synthase proteins.

3. The cereal flour composition according to claim 1 that contains 1 to 30 wt % of the wheat flour prepared from a type of wheat that does not express any two of the three wheat starch synthase II proteins, and also does not express the three wheat granule bound starch synthase proteins.

4. The cereal flour composition according to claim 1 that contains 2 to 20 wt % of the wheat flour prepared from a type of wheat that does not express any two of the three wheat starch synthase II proteins, and also does not express the three wheat granule bound starch synthase proteins.

5. The cereal flour composition according to claim 1, wherein the wheat flour prepared from a type of wheat that does not express any two of the three wheat starch synthase II proteins, and also does not express the three wheat granule bound starch synthase proteins is a wheat flour prepared from a type of wheat that does not express the wheat starch synthase II-A1 protein and the wheat starch synthase II-B1 protein among the three wheat starch synthase II proteins, and also does not express the three wheat granule bound starch synthase proteins.

6. The cereal flour composition according to claim 1, wherein the other cereal flour is selected from the group consisting of wheat flour from non-waxy wheat, starch, rice flour, buckwheat flour, barley flour, corn flour, and oat flour.

7. A process for producing a food product, comprising mixing auxiliary food ingredients with the cereal flour composition according to claim 1.

8. A food product manufactured by mixing auxiliary food ingredients with the cereal flour composition according to claim 1.

9. The food product according to claim 8, wherein the food product is selected from the group consisting of bakery goods, noodles, deep fried foods, foods cooked on a griddle, roux and sauces.