Method for breeding mianrou type distiller's yeast peavariety and application thereof

Abstract

The application provides a breeding method and application of a soft type liquor koji making pea variety, and relates to the technical field of soft type liquor making raw material breeding.The application provides a breeding method of a soft type liquor koji making pea variety, which comprises the following steps: crossing Su Gua No.3 as a female parent and an unnamed pea No.7 as a male parent to obtain F1 generation; continuously performing multiple generations of self-cross breeding from the F1 generation until a pea line with stable grain variety indexes is obtained; then making koji with the pea line, and screening the soft type liquor koji making pea variety according to koji making product indexes.The breeding method can be adapted to planting in three core ecological zones in Jiangsu, improve the cold resistance, waterlogging resistance and heat resistance of the pea variety, significantly improve the yield in different ecological zones, has excellent yield stability, and improves the overall stability of koji making, and breaks the technical barrier between breeding and the liquor making industry.

Description

Technical Field

[0001] This invention belongs to the field of breeding technology for raw materials for mellow-type baijiu brewing, specifically involving a method for breeding and applying a pea variety for mellow-type baijiu koji making. Background Technology

[0002] Mellow-style baijiu, with Daqu (a type of starter culture) as its core, is characterized by its smooth entry, lack of harshness, and long-lasting aftertaste. The Daqu used in mellow-style baijiu is generally made at a medium temperature. Peas are a core ingredient in the quality and style of mellow-style baijiu Daqu, playing an irreplaceable triple core role in the Daqu brewing system: Nutritional supply function: Its rich protein can be hydrolyzed by the protease secreted by mold during the koji-making process to generate a variety of amino acids, providing a high-quality nitrogen source for yeast fermentation; its starch can provide the main carbon source for alcoholic fermentation after saccharification and degradation.

[0003] Structural regulation: The unique viscous proteins and polysaccharides in peas can optimize the physical structure of the koji blank, balance its water retention and air permeability, stabilize the temperature rise and fall rhythm of the koji, and create a microenvironment inside the koji suitable for the stable growth of ester-producing microorganisms such as Rhizopus, Aspergillus, and Mucor.

[0004] Flavor contribution: The aromatic amino acids such as phenylalanine and tyrosine produced by the decomposition of pea protein can generate complex aroma substances such as ethyl acetate and ethyl hexanoate through Maillard reaction, laying the foundation for the unique aroma and taste of mellow baijiu.

[0005] However, existing pea varieties suffer from three major defects: poor stability in koji production, uneven cold, waterlogging, and heat resistance, and poor yield and flavor in mellow-style baijiu. These defects have become the core technical bottleneck restricting the quality stability of mellow-style baijiu koji. Among them, Suwan No. 3 has excellent nutritional indicators, but its seeds have a low content of sticky substances. During koji production, the excessive aeration of the koji blanks leads to rapid moisture loss and insufficient post-fermentation, easily resulting in quality problems such as underdeveloped koji blanks and excessively rapid annealing. Fanglv No. 1 has excellent commercial traits, but as a fresh-eating variety, its moisture content and dry-basis starch content cannot meet the raw material requirements for baijiu koji production. This creates a technical information barrier between the breeding and brewing fields, resulting in the industry problem of a disconnect between raw material breeding and baijiu koji production processes. Therefore, there is an urgent need to provide a new pea variety breeding method specifically adapted to the mellow-style baijiu koji production process to break down the technical barriers between the breeding and brewing industries. Summary of the Invention

[0006] To address the aforementioned technical problems, the primary objective of this invention is to provide a method for breeding pea varieties for producing mellow-tasting baijiu (Chinese liquor). This method is adaptable to planting in the three core ecological zones of Jiangsu Province, improves the cold resistance, flood resistance, and heat resistance of pea varieties, significantly increases yields in different ecological zones, exhibits excellent yield stability, and enhances the overall stability of koji production, thus breaking down technical barriers in the breeding and brewing industries.

[0007] The second objective of this invention is to provide the application of the above-mentioned breeding method in screening cold-resistant, flood-resistant and heat-resistant pea varieties.

[0008] A third objective of this invention is to provide the application of the above-described breeding method in screening high-yield pea varieties.

[0009] The fourth objective of this invention is to provide the application of the above-mentioned breeding method in improving the quality and / or stability of Daqu (a type of starter culture) for mellow-type Baijiu.

[0010] To achieve the above-mentioned objectives, the technical solution of this invention is as follows: This invention provides a method for breeding pea varieties for making mellow-type baijiu (Chinese liquor) starter culture, comprising the following steps: using Suwan No. 3 as the female parent and Wumingwan No. 7 as the male parent to cross and obtain the F1 generation; starting from the F1 generation, continuously carry out multiple generations of self-pollination and breeding until a pea line with stable seed varietal indicators is obtained; then, the pea line is used to make starter culture, and mellow-type baijiu starter culture pea varieties are selected based on the indicators of the starter culture product. The grain varietal indicators include crude protein content, crude starch content, and 20-mesh fine flour ratio; The product indicators of the koji-making process include the saccharification power, fermentation power, saccharification power / fermentation power ratio, and koji-forming rate of the koji blank.

[0011] In one implementation, the multi-generational self-pollination breeding consists of 5 generations, namely F2, F3, F4, F5 and F6.

[0012] In one embodiment, the crude protein content is 18-24%, the crude starch content is ≥45%, and the 20-mesh fine powder rate is 20-32%.

[0013] In one embodiment, the saccharification power of the Daqu (a type of starter culture) is ≥600 mg glucose / (g·h), the fermentation power of the Daqu is ≥1.2 g CO2 / (g·48 h), the saccharification power / fermentation power ratio is 2.0~3.5, and the forming rate of the starter culture is ≥95%.

[0014] In one implementation method, the conditions for making koji are: temperature of 30-35°C and humidity of 70%-80% for days 1-4; and temperature of 40-45°C and humidity of 60%-70% for days 5-12.

[0015] In one implementation method, the weight ratio of wheat, barley and pea seed lines in the koji-making process is (4~6):(3~5):(0.5~1.5).

[0016] As one implementation method, the product indicators for koji production also include the coefficient of variation of output in different regions, wherein the coefficient of variation of output in different regions is ≤5%, and the number of different regions is 3 or more.

[0017] The present invention also provides the application of the above-mentioned breeding method in screening cold-resistant, flood-resistant and heat-resistant pea varieties.

[0018] The present invention also provides the application of the above-mentioned breeding method in screening high-yield pea varieties.

[0019] The present invention also provides the application of the above-mentioned breeding method in improving the quality and / or stability of Daqu (a type of starter culture) for mellow-type Baijiu.

[0020] The advantages of this invention compared to existing technologies are as follows: This invention provides a method for breeding pea varieties for producing mellow-type baijiu (Chinese liquor) koji (fermentation starter). This method integrates seed variety indicators with koji product indicators into the pea breeding evaluation system, achieving a technological upgrade from general grain breeding to targeted breeding specifically for brewing koji, breaking down technical barriers between the breeding and brewing industries. This invention specifically targets the complex climate of Jiangsu, characterized by "plum rain-high temperature-cold humidity," and the bred varieties are adaptable to planting in Jiangsu's three core ecological zones, exhibiting excellent yield stability, with dry seed yields reaching 180-200 kg per mu (approximately 0.067 hectares), representing an increase of over 10%, and an annual yield variation coefficient ≤15%. The saccharification power variation coefficient of the mellow-type baijiu koji pea varieties obtained through this method is reduced from 18%-25% of traditional varieties to below 10%, the koji formation rate is increased from 85% to over 95%, and the overall stability of koji production is improved by over 10%, addressing the industry pain point of large batch-to-batch quality fluctuations in mellow-type baijiu from the source. Detailed Implementation

[0021] This invention provides a method for breeding pea varieties for making mellow-type baijiu (Chinese liquor) starter culture, comprising the following steps: using Suwan No. 3 as the female parent and Wumingwan No. 7 as the male parent to crossbreed to obtain the F1 generation; starting from the F1 generation, continuously carry out multiple generations of self-pollination and breeding until a pea line with stable seed varietal indicators is obtained; then, the pea line is used to make starter culture, and mellow-type baijiu starter culture pea varieties are selected based on the starter culture product indicators of the starter culture.

[0022] This invention selects Suwan 3 as the female parent. Suwan 3 is a variety bred by the Jiangsu Yangtze River Agricultural Research Institute. It has a spring-sown growth period of 80 days, is a medium-maturing variety, has a crude starch content of 46.83%, and exhibits excellent basic quality for koji making. This invention selects the stress-resistant donor Wumingwan 7 as the male parent. Wumingwan 7 is a local northern germplasm, scientifically known as... Pea Sativum L. The unified number is G0000337, published in the China Crop Germplasm Information Network. It has excellent drought resistance, salt tolerance, disease resistance and cold resistance characteristics, and can effectively improve the ecological adaptability of offspring varieties.

[0023] In this invention, hybrid seeds are obtained by crossbreeding Suwan No. 3 and an unnamed pea variety No. 7, both characterized by plump grains, bright color, and freedom from discoloration, mold, and insect infestation. The F1 generation is then selected based on seed variety indicators, including crude protein content, crude starch content, and 20-mesh fine flour percentage. In this invention, the crude protein content is 18-24%, the crude starch content is ≥45%, and the 20-mesh fine flour percentage is 20-32%. Optionally, the crude protein content is determined according to NY / T 3-1982 "Determination of Crude Protein in Cereal and Legume Seeds (Semi-micro Kjeldahl Method)," and the crude starch content is determined according to NY / T11-1985 standard method. The pea seeds are ground into powder using common equipment such as a pulverizer, and the 20-mesh fine flour percentage is determined according to "Grain and Oil Inspection: Determination of Fineness of Flour" (GB / T 5507-2008).

[0024] This invention involves continuous self-pollination across multiple generations starting from the F1 generation until a pea line with stable seed quality indicators is obtained. In this invention, the multi-generation self-pollination involves five generations: F2, F3, F4, F5, and F6. From each generation, this invention selects individual plants exhibiting strong disease resistance, high yield per plant, and vigorous growth, and then screens them based on seed quality indicators, including crude protein content, crude starch content, and 20-mesh fine flour percentage. In this invention, the crude protein content is 18-24%, the crude starch content is ≥45%, and the 20-mesh fine flour percentage is 20-32%. Ultimately, superior individual plants with excellent agronomic traits, meeting starch and protein standards, and achieving a satisfactory 20-mesh fine flour percentage are obtained. This invention establishes a three-dimensional evaluation index system based on the adaptability of koji-making process, encompassing starch characteristics, protein characteristics, and pulverization adaptability. Starch index screening identifies the minimum process requirement of saccharification power ≥600 mg glucose / (g·h) for mellow-type baijiu koji. Protein index achieves a balance between enzyme production by molds and nitrogen supply by yeast; levels below 18% result in insufficient enzyme production, while levels above 24% easily produce off-flavors, affecting the mellow style of the liquor. Pulverization adaptability index optimizes the ratio of "skin, grits, and flour" in the koji blank, achieving a balance between air permeability and water retention, ensuring a stable koji-making process, and identifying pea varieties suitable for koji-making.

[0025] In this invention, the pea strain is then used to make koji. The koji making process of this invention is carried out in accordance with the mellow type of baijiu preparation process. The mellow type of baijiu preparation process of this invention includes the following steps: wheat, barley and peas are mixed in a weight ratio of (4~6):(3~5):(0.5~1.5), crushed and mixed with water, and then pressed into koji blanks for fermentation and cultivation.

[0026] In this invention, the preferred weight ratio of wheat, barley, and peas is 5:4:1. The raw materials are mixed and then pulverized to obtain a mixture, which is passed through a 30-60 mesh sieve, preferably a 40-50 mesh sieve. After pulverization, water is added to the mixture and mixed thoroughly. The weight ratio of the mixture to water is (1.5-2.5):1, preferably 2:1. After mixing, the mixture is pressed into koji blanks, with a koji blank forming rate ≥95%. The koji blank forming rate = (number of koji blocks meeting appearance and size standards / total number of pressed koji blocks) × 100%. The koji blanks are pressed until they do not crumble when left to stand naturally, and then fermented to complete the koji production. In this invention, the fermentation time is 12 days; the fermentation conditions are: temperature 30-35℃ and humidity 70%-80% for days 1-4; and temperature 40-45℃ and humidity 60%-70% for days 5-12.

[0027] After the koji (fermentation starter) is made in this invention, pea varieties for making mellow-type baijiu (Chinese liquor) are selected based on the koji product indicators. In this invention, the koji product indicators include the saccharification power, fermentation power, and saccharification power / fermentation power ratio of the koji blank. Specifically, the saccharification power, fermentation power, and saccharification power / fermentation power ratio of the koji are determined and calculated according to QB / T 4257-2011 "General Analytical Methods for Brewing Koji". In this invention, pea varieties for making mellow-type baijiu are selected based on the saccharification power, fermentation power, saccharification power / fermentation power ratio, and koji blank formation rate, wherein the saccharification power of the koji is ≥600 mg glucose / (g·h), the fermentation power is ≥1.2 g CO2 / (g·48 h), the saccharification power / fermentation power ratio is 2.0~3.5, and the koji blank formation rate is ≥95%.

[0028] In this invention, the product indicators for koji production also include the coefficient of variation in yield across different regions, and the method for calculating the coefficient of variation in yield is as follows: 150 (%) = Standard deviation (%) S ) / average value( In the formula, S and The standard deviation and average yield of pea varieties at various regional test sites are given; the coefficient of variation of yield in different regions is ≤5%, and the number of different regions is more than 3. In this invention, the different regions include the Yangtze River area of ​​Nantong, Jiangsu Province, the Lixiahe area of ​​Yangzhou, and the Huanghuai area of ​​Suqian.

[0029] This invention also provides the application of the above-described breeding method in screening cold-resistant, flood-resistant, and heat-resistant pea varieties. The pea varieties screened by the breeding method of this invention possess cold-resistant, flood-resistant, and heat-resistant characteristics, and can be used to screen for cold-resistant, flood-resistant, and heat-resistant pea varieties.

[0030] This invention also provides the application of the above-described breeding method in screening high-yielding pea varieties. The pea varieties screened using the breeding method of this invention exhibit significantly increased yields and can be used for screening high-yielding pea varieties.

[0031] This invention also provides the application of the above-mentioned breeding method in improving the quality and / or stability of mellow-type baijiu koji (fermentation starter). The pea varieties screened by the breeding method of this invention improve the quality and stability of mellow-type baijiu koji after koji production, and can be used to improve the quality and / or stability of mellow-type baijiu koji. In this invention, the quality of mellow-type baijiu koji includes yield, mellowness, and purity. The stability refers to the consistency of the quality of mellow-type baijiu koji prepared from peas from different geographical regions.

[0032] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be described in detail below with reference to the embodiments, but these should not be construed as limiting the scope of protection of this invention.

[0033] Unless otherwise specified, the materials, reagents, etc. used in the following examples are commercially available. Unless otherwise specified, they are generally used under conventional conditions or under conditions recommended by the company.

[0034] Suwan No. 3: Spring sowing growth period of 88 days, crude starch 46.83%, crude protein 21.47%, strong lodging resistance.

[0035] Unnamed Pea No. 7: Spring-sown, with a growth period of 81 days, crude starch content of 49.12%, crude protein content of 25.33%, and excellent disease resistance, cold resistance, and drought resistance.

[0036] Experimental Example 1 1. Field phenotypic screening Crude starch content was determined according to the standard method of NY / T11-1985, and crude protein content was determined according to NY / T 3-1982 "Determination of Crude Protein in Cereal and Legume Seeds (Semi-micro Kjeldahl Method)". The 20-mesh fine powder ratio was determined by grinding pea seeds into powder using common pulverizing equipment and according to "Grain and Oil Inspection: Determination of Fineness of Flour" (GB / T 5507-2008).

[0037] In the spring of 2021, Suwan No. 3, a variety with plump, bright-colored seeds free from discoloration, mold, and insect infestation, was selected as the female parent, and an unnamed pea variety No. 7 as the male parent for hybridization. An experimental field was set up in Siyang County, Suqian City, Jiangsu Province to conduct germplasm resource screening. Each experimental population consisted of no fewer than 2000 individual plants, using a randomized block design with three biological replicates. The parents were isolated by striping at the early flowering stage, and artificial pollination was performed during the flowering and maturity period to complete hybridization. Hybrid seeds were harvested after the pods matured. Then, using a three-dimensional evaluation index of crude starch, crude protein, and 20-mesh fine flour content, 23 F1 generation seeds with a crude starch content ≥45%, crude protein content 18%–24%, and 20-mesh fine flour content 20%–32% were obtained from the hybrid seeds.

[0038] From winter 2021 to spring 2023, 23 F1 generation seeds were used as parents for self-pollination. A population of 1100 vigorous plants with strong lodging and disease resistance, a single-plant yield ≥15 g, excellent grain uniformity, a thousand-grain weight ≥220 g, and a plant height of 60-80 cm were selected as the F2 generation. The F2 generation was then used as parents for continuous self-pollination in Suqian and Hainan until the F6 generation. From the F3, F4, F5, and F6 generations, trait screening was conducted based on a three-dimensional evaluation index system: crude starch content ≥45%, crude protein content 18%-24%, and 20-mesh fine flour rate 20%-32%. Ultimately, 57 superior plants with excellent agronomic traits, meeting starch and protein standards, and a qualified 20-mesh fine flour rate were retained, representing an overall retention rate of 5.2%. These 57 superior plants exhibited characteristics such as a plant height of 65-75 cm, good lodging resistance, stable pod-setting traits, and high grain uniformity.

[0039] 2. Re-screening during koji-making process In the autumn of 2023, a second screening was conducted to assess the compatibility of F6 generation pea varieties with Daqu (fermentation starter) prepared using the standardized process for producing mellow-style baijiu. A 28-day Daqu-making experiment was carried out simulating the local climate conditions of Jiangsu province: 57 stable varieties were prepared using the standardized Daqu-making process for mellow-style baijiu, with three replicates for each variety and 50 kg of Daqu material per replicate. The temperature was maintained at 30-35℃ and relative humidity at 70%-80% during the early stage of Daqu production; the temperature was maintained at 40-45℃ and relative humidity at 60%-70% during the later stage of Daqu production, and the mixture was cultured under constant temperature and humidity for 28 days.

[0040] After the koji making process was completed, the stability of each koji strain was evaluated using the following indicators: saccharification power of 685 mg glucose / (g·h), fermentation power of 1.45 g CO2 / (g·48h), saccharification power / fermentation power ratio of 2.38, and koji formation rate. Three stable strains with excellent koji-making adaptability were selected. Among them, the strain "Yangmianwan No. 1" showed the best overall performance, with a saccharification power of 685 mg glucose / (g·h), fermentation power of 1.45 g CO2 / (g·48h), saccharification power / fermentation power ratio of 2.38, and a koji formation rate of 98%, with no quality defects such as bulging, underdeveloped core, or cracks.

[0041] Experimental Example 2 From 2024 to 2025, a multi-site regional trial of "Yangmianwan No. 1" was conducted for two consecutive years in three typical ecological zones: the Yangtze River Delta region of Nantong, the Lixiahe region of Yangzhou, and the Huanghuai region of Suqian, Jiangsu Province. Each trial site had at least three replicates. Using Suwan No. 3 as the control variety, unified field management standards were applied at all trial sites. The yield stability and koji-making stability of each variety were comprehensively evaluated. The results are shown in Table 1 below. Table 1. Results of yield and koji-making stability in different regions

[0042] The experimental results show that "Yangmianwan No. 1" can grow and bear fruit normally in the three major ecological zones of Jiangsu Province, with stable stress resistance and a yield variation coefficient of only 4.0%. For two consecutive years, all koji-making suitability indicators have been within the qualified threshold range. It has excellent stable yield in the three major ecological zones, and the saccharification power variation coefficient of koji is 8.7%, which is much better than that of the control variety Suwan No. 3. The raw material yield and koji quality stability have been greatly improved, and it fully meets all the requirements of the national standard GB / T10460—2008 "Pea".

[0043] Experimental Example 3 In the winter of 2025, pilot production verification was carried out on the production line of a mellow-style baijiu production enterprise in Jiangsu. Special koji was prepared using "Yangmianwan No. 1" peas and "Suwan No. 3" peas and used for baijiu brewing. Specifically, wheat, barley and Yangmianwan No. 1 peas (or Suwan No. 3 peas) were mixed in a weight ratio of 5:4:1 and crushed through a 40-mesh sieve. After crushing, water was added to the mixture in a weight ratio of 2:1 and mixed evenly. Then, the mixture was pressed into koji blanks. Fermentation and cultivation were carried out under the conditions of 30-35℃ and 70%-80% humidity from day 1 to day 4, and 40-45℃ and 60%-70% humidity from day 5 to day 12 to complete the koji making. Production results show that, compared with the Suwan No. 3 pea raw material, the "Yangmianwan No. 1" of this invention increases the alcohol yield of mellow-type baijiu from 29.8% to 30.5%, an increase of 2.3%, and the premium alcohol yield from 23.6% to 24.9%, an increase of 5.6%. It also effectively reduces the content of fusel alcohols in the liquor, with isoamyl alcohol decreasing from 263.5 mg / L to 242.3 mg / L and n-butanol decreasing from 582.6 mg / L to 528.1 mg / L. The mellowness and purity of the liquor are significantly improved, and the sensory score increases from 95.3 to 98.8, an increase of 3.5 points (out of 100). The style characteristics of mellow-type baijiu are more prominent and stable.

[0044] In summary, the bred varieties obtained by the breeding method of this invention fully comply with the national standard GB / T10460—2008 "Pea". The appearance, moisture and impurities meet the standards. Without modifying the existing koji-making equipment and production process, it can be seamlessly connected to the existing mellow-type baijiu koji-making production supply chain, improve the mellowness and purity of the liquor, and make the style characteristics of mellow-type baijiu more prominent and stable.

[0045] The above description is merely an embodiment of the present invention and does not limit the patent scope of the present invention. Any equivalent structural or procedural transformations made based on the content of the present invention specification, or direct or indirect applications in other related technical fields, are similarly included within the patent protection scope of the present invention.

Claims

1. A method for breeding a pea variety for making mellow-tasting baijiu (Chinese liquor) starter, characterized in that, Includes the following steps: The F1 generation was obtained by crossing Suwan No. 3 as the female parent and Wumingwan No. 7 as the male parent; Starting from F1, multiple generations of self-pollination and breeding were carried out until a pea line with stable seed varietal indicators was obtained; then, the pea line was used to make koji, and pea varieties for making mellow baijiu were selected based on the indicators of the koji product. The grain varietal indicators include crude protein content, crude starch content, and 20-mesh fine flour ratio; The product indicators of the koji-making process include the saccharification power, fermentation power, saccharification power / fermentation power ratio, and koji-forming rate of the koji blank.

2. The breeding method according to claim 1, characterized in that, The multi-generation self-pollination breeding consists of 5 generations, namely F2, F3, F4, F5 and F6.

3. The breeding method according to claim 1, characterized in that, The crude protein content is 18-24%, the crude starch content is ≥45%, and the 20-mesh fine powder rate is 20-32%.

4. The breeding method according to claim 1, characterized in that, The saccharification power of the Daqu (a type of starter culture) is ≥600 mg glucose / (g·h), the fermentation power of the Daqu is ≥1.2 g CO2 / (g·48 h), the saccharification power / fermentation power ratio is 2.0~3.5, and the forming rate of the starter culture is ≥95%.

5. The breeding method according to claim 1, characterized in that, The conditions for making koji are: temperature 30-35℃ and humidity 70%-80% for days 1-4; and temperature 40-45℃ and humidity 60%-70% for days 5-12.

6. The breeding method according to claim 1, characterized in that, The weight ratio of wheat, barley and pea varieties in the koji-making process is (4~6):(3~5):(0.5~1.5).

7. The breeding method according to any one of claims 1 to 6, characterized in that, The product indicators for koji making also include the coefficient of variation of output in different regions, wherein the coefficient of variation of output in different regions is ≤5%, and the number of different regions is 3 or more.

8. The application of the breeding method according to any one of claims 1 to 7 in screening cold-resistant, flood-resistant and heat-resistant pea varieties.

9. The application of the breeding method according to any one of claims 1 to 7 in screening high-yield pea varieties.

10. The application of the breeding method according to any one of claims 1 to 7 in improving the quality and / or stability of Daqu (a type of starter culture) for mellow-style Baijiu.

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