CN110999781A

CN110999781A - Method for rapidly screening high-quality beer barley resistant to barley yellow mosaic disease

Info

Publication number: CN110999781A
Application number: CN201911271554.4A
Authority: CN
Inventors: 徐肖; 沈会权; 栾海业
Original assignee: JIANGSU LITTORAL AGRICULTURE SCIENCE RESEARCH INSTITUTE
Current assignee: JIANGSU LITTORAL AGRICULTURE SCIENCE RESEARCH INSTITUTE
Priority date: 2019-12-02
Filing date: 2019-12-02
Publication date: 2020-04-14

Abstract

The invention relates to a method for rapidly screening high-quality beer barley resistant to barley yellow mosaic disease, which comprises the following steps: 1) collecting high-quality malting barley varieties (lines) for planting, investigating the disease resistance level of malting barley, examining the plant height (cm), the ear length (cm), the grain number (grains) of a single ear, the grain number (grains) of a single plant, the grain weight (g) of a single plant and the thousand grain weight (g), and measuring the Korla value (%); 2) and (4) carrying out correlation analysis on 6 agronomic traits and disease grades of the n germplasms, and researching the correlation among the traits. 3) And (3) performing cluster analysis and principal component analysis by taking 8 main traits as indexes, calculating an M value, performing comprehensive evaluation on the barley germplasm of the beer, and screening the germplasm with excellent comprehensive traits. The method is relatively comprehensive, easy to scale, program and informationize, and lays a foundation for the application of the remote sensing measurement technology in barley germplasm screening and identification of high-generation breeding materials.

Description

Method for rapidly screening high-quality beer barley resistant to barley yellow mosaic disease

Technical Field

The invention relates to a method for rapidly screening high-quality beer barley resistant to barley yellow mosaic disease, and belongs to the technical field of agriculture.

Background

The coastal region of Jiangsu has flat terrain, mild climate, moderate rainfall and sufficient illumination, the unique ecological environment is particularly suitable for the production of beer barley, the production area of the three beer barley in China is one of the three, the annual planting of the beer barley is about 150 ten thousand mu, and the produced beer barley accounts for more than one third of the supply of the domestic beer barley. The yellow mosaic disease of barley occurs in large area in the middle and lower reaches beer barley production area of Yangtze river in China, and is the most important virus disease affecting the yield and quality of beer barley.

The beer barley germplasm resource is the basis of beer barley germplasm innovation and biological research, reasonable multivariate analysis and evaluation are carried out on the genetic diversity of the beer barley germplasm resource, the development of high-quality beer barley germplasm resisting barley yellow mosaic disease is facilitated, the excellent genes of the beer barley germplasm resource are fully utilized, the unicity of breeding backbone parents is prevented, and the breeding process is accelerated.

The genetic diversity of barley germplasm has been studied at different levels by phenotypic trait identification, molecular markers, biochemical markers, and the like. The evaluation method of the phenotypic character is most intuitive and is easy to master by breeders, and because different properties of the barley are related to each other to different degrees, people pay attention to how to comprehensively evaluate the barley germplasm by using multiple properties. Zhou Wei utilizes the principal component to analyze and screen important components of agronomic characters of Erlenmeyer barley germplasm, obtains specific germplasm by a systematic clustering method, and can be used as breeding parents or intermediate materials of different types (Zhou Wei, Hebei agricultural science). The method is characterized in that the poplar bright identifies and clusters the agronomic characters of the malting barley varieties in different regions at home and abroad, and the diversity distribution of the malting barley in different regions is considered to have obvious ecological regionality (poplar bright, a plant genetic resource bulletin). In recent years, two-dimensional sequencing analysis based on main components is gradually introduced into germplasm identification and comprehensive evaluation of crops such as hemp, garlic, wheat, cotton and the like, and character performances of various qualities and differences among the qualities are revealed, and the analysis method is not reported in beer barley. The method is characterized in that the multi-character of the germplasm is comprehensively evaluated by combining the principal component analysis and the multiple linear regression construction of the evaluation equation of the core germplasm of the crops such as millet, rice, upland cotton and the like, but the method is not applied to the germplasm evaluation of beer barley.

Disclosure of Invention

The invention aims to provide a technology for rapidly screening quality barley malus domestica germplasm for resisting malus domestica mosaic disease, and aims to establish a simple, feasible, stable and reliable method for rapidly screening malus domestica mosaic disease-resistant beer barley with excellent comprehensive properties.

The invention adopts the following technical scheme:

the test uses n selected beer barley varieties (lines) from home and abroad as test materials, and the materials are planted in a barley yellow mosaic disease identification garden for many years by adopting a random block design, wherein each material is sown with 1 row, the row length is 1.2m, the row spacing is 0.2m, the plant spacing is 3cm, each row is sown with 40 grains, and the steps are repeated for 3 times.

And (4) performing barley yellow mosaic disease resistance investigation in 2-3 months of the next year, wherein the investigation method refers to the standard of Huangpeloyal and the like. After the seeds are mature, 6 agronomic characters of plant height (cm), spike length (cm), single spike number (grain), single plant grain weight (g) and thousand grain weight (g) are examined, and 1 quality character of a Kulbah value (%) is measured.

And (3) carrying out clustering and principal component analysis by using SPSS 21.0 statistical software, wherein in the clustering process, the inter-germplasm distance is Euclidean distance, and the clustering method adopts a dispersion square sum method. The total nitrogen and soluble nitrogen contents of the malt were measured by Kjeldahl method.

6 agronomic character phenotypes of the planted n beer barley germplasms are analyzed, genetic diversity indexes of the n beer barley germplasms are researched, 6 agronomic characters and disease levels of the n beer barley germplasms are subjected to related analysis, and the correlation among main characters of the n beer barley germplasms is researched.

And (3) performing cluster analysis on the planted n Erlenmeyer barley germplasms by using R language based on a Mahalanobis distance class average method by taking 8 main traits as indexes. In order to eliminate the influence of correlation among characters, the germplasm evaluation is carried out by adopting principal component analysis. And extracting the first N main components according to the principle that the accumulated contribution rate is greater than 70%. Carrying out standardization treatment on 8 main character numerical values of the beer barley germplasm resources to obtain first N principal component scores, carrying out normalization treatment on the N principal component scores, calculating each principal component weight coefficient according to the contribution rate, and carrying out comprehensive evaluation on various materials by using the sum M value of the products of the score values of the first N principal components and the corresponding weights as a comprehensive index, wherein the higher the M value is, the better the comprehensive character performance is, and the germplasm with excellent comprehensive characters is screened out.

The invention is relatively comprehensive, easy to scale, program and informationize, and lays a foundation for the application of the remote sensing measurement technology in beer barley germplasm screening and high-generation breeding material screening and identification.

Drawings

FIG. 1 beer barley germplasm cluster map

Detailed Description

The following examples are provided to illustrate the practice of the present invention.

Example (b):

86 collected and screened malting barley varieties (lines) from home and abroad are taken as test materials in the research. The test materials are planted in the Nanyang test farm of the agricultural institute in the coastal region of Jiangsu in autumn sowing 2017, a random block design is adopted, 1 row of materials is sown, the row length is 1.2m, the row spacing is 0.25m, the plant spacing is 3cm, 40 seeds are sown in each row, and the sowing is repeated for 3 times. And planting the test material on a Nanyang test farm of an agricultural institute in coastal areas of Jiangsu in autumn sowing in 2018, planting the test material in the same specification in 2017, and performing conventional cultivation management.

The barley yellow mosaic disease resistance is investigated in 2018 in 2-3 months, and the disease grade is divided into four grades of 1, 2, 3 and 4 according to the investigation method by referring to the standard of Huangpeloy and the like to evaluate the barley yellow mosaic disease resistance:

grade 1-normal green leaves without spots;

level 2-leaf color is basically normal with yellow spots, but spots are not connected into lines;

grade 3-spots of leaf lesions are connected into lines, the leaves are yellow, but the plants are not dwarfed;

grade 4-large yellow spots appear on leaves, leaves yellow, plants wither, dwarfed and even die.

And investigating the disease level in the same way in months 2-3 in 2019.

In 2018, after the barley is mature, 5 competitive plants are taken from each material, and 6 agronomic characters such as plant height (cm), spike length (cm), single spike grain number (grain), single plant spike number (grain), single plant grain weight (g) and thousand grain weight (g) are recorded according to barley germplasm resource description standards and data standards. 6 traits were investigated in the same way in 5 months in 2019.

The total nitrogen and soluble nitrogen contents of the malt were measured by Kjeldahl method.

The 86 beer barley germplasms are analyzed for 6 main agronomic character phenotypes, genetic diversity indexes are calculated, and the analysis results are listed in table 1.

TABLE 1 analysis of genetic diversity index of main traits in barley germplasm resources for beer

6 agronomic characters H' of the reference material show that the genetic diversity index of the spike length is the highest (2.05), the genetic diversity index of the thousand seed weight is the lowest (1.87), and the amplitude variation is 1.87-2.05. The genetic diversity of the beer barley germplasm resources is relatively rich.

The correlation analysis results among 7 main traits of 86 parts of beer barley germplasm are shown in a table 2, and the table shows that the plant height is extremely obviously positively correlated with the ear length, the thousand grain weight, the single ear grain number and the single plant grain weight average, and is extremely obviously negatively correlated with the disease grade; the ear length is in extremely obvious positive correlation with the grain number of a single ear, the grain number of a single plant, the thousand grain weight, the grain weight of a single plant and the disease grade; the single ear number is in extremely obvious negative correlation with the single plant ear number, the thousand grain weight and the single plant weight average, and is in extremely obvious positive correlation with the disease grade; the number of the single plant ears is in extremely obvious positive correlation with thousand grain weight and in extremely obvious negative correlation with disease grade; thousand seed weight is in extremely obvious positive correlation with single plant seed weight, is in extremely obvious negative correlation with disease grade, and single plant seed weight is in extremely obvious negative correlation with disease grade.

TABLE 2 correlation of the major traits of Erlenmeyer's wort germplasm resources

Note: and represent significance at the 0.05 and 0.01 levels, respectively.

The 86 beer barley germplasms were subjected to cluster analysis using R language based on the Mahalanobis distance's class-averaging method using 8 major traits as indices, and the results are shown in FIG. 1. 86 Erlenmeyer germplasm can be grouped into 4 classes at Mahalanobis distance 10.

The I category comprises 5 germplasms, 1430R, Yangyuan 02, Zhou Chong No. 1, high-quality No. 1 and Hu 01-2946, and is characterized by high stalk, long spike, moderate spike grain number, more single plant spike, single plant grain weight, thousand grain weight and Kulbar Ha value, and general comprehensive character performance.

The II class comprises 20 germplasm which is respectively from 9 parts of Jiangsu, 4 parts of Henan, 2 parts of Hubei, 2 parts of Zhejiang, 1 part of Shanghai, 1 part of Sichuan and 1 part of Fujian, and mainly breeds popularized in each province in recent years. The germplasm is characterized by short stalk, more single plant ears, higher thousand grain weight and single plant grain weight, moderate ear length, moderate Kulbah value and better comprehensive performance.

The III category comprises 24 germplasm, 7 parts of Jiangsu, 5 parts of Hubei, 2 parts of Zhejiang, Yunnan, Heilongjiang and Henan germplasm, 1 part of Shaanxi and Fujian germplasm, 2 parts of Japan and 1 part of UK germplasm. The germplasm is characterized by moderate plant height, more grains per spike, single plant, thousand grain weight and single plant grain weight, shorter spikes, higher Kulbah value, and moderate comprehensive performance, which is close to the first analogy.

The IV class comprises 37 parts of germplasm, which is mainly spring barley, and has the advantages of plant height, spike length, central spike number of each plant, lower thousand grain weight, less spike grain number, low grain weight of each plant, higher Kulbah value and poorer comprehensive performance in Jiangsu ecological conditions.

In order to eliminate the influence of correlation among characters, the germplasm evaluation is carried out by adopting principal component analysis. According to the principle that the cumulative contribution rate is more than 70%, most of biological information of all characters can be summarized by extracting the first 4 main components (Table 3).

The contribution rate of the first main component is 32.80%, the characteristic vector values of plant height, ear length, single ear grain number, thousand grain weight and single plant grain weight are the largest and are positive values, the condensed biological information is directly related to yield and can be summarized as a yield factor, the ear length, the ear grain number increase and the grain weight increase are beneficial to high yield, and the first main component is good.

The contribution rate of the second principal component is 23.30%, and the value of the characteristic vector of the disease grade is the largest and is a negative value; the number of ears per plant is large and positive. The biological information is summarized as a resistance factor, and the second principal component should be properly biased to be better.

The contribution rate of the third main component is 15.98%, the characteristic vector value of the number of ears per plant is the largest and positive value, so the biological information covered by the main component is the tillering factor, and according to the relationship of the three factors of yield, the increase of the number of ears per plant affects the improvement of thousand kernel weight and the number of ears per plant, and the disease grade is also positive value, so the third main component should be slightly smaller.

The contribution rate of the fourth principal component is 12.17%, the characteristic vector value of the Kolbar-Ha value is the largest and positive value, biological information of the fourth principal component is summarized as the quality factor, the Kolbar-Ha value which is too high or too low affects the filtration of yeast and wort, the stability of foam and the flavor of beer to different degrees, generally, the Kolbar-Ha value of malt should be kept around 40% -46%, and the fourth principal component should be moderate and good.

TABLE 3 eigenvalues and contribution rates of germplasm resources of Erlenmeyer barley

The method comprises the steps of standardizing 8 main agronomic character numerical values of the germplasm resources of the Erlenmeyer barley to obtain the first 4 principal component scores, normalizing the 4 principal component scores, calculating the weight coefficients of the principal components to be 0.3893, 0.2766, 0.1897 and 0.1444 respectively according to the contribution rate, and comprehensively evaluating various materials by using the sum M value of the products of the scores of the first 4 principal components and the corresponding weights as a comprehensive index, wherein the higher the M value is, the better the comprehensive character performance is. The Erlenmeyer barley germplasm materials arranged in the first ten positions are respectively as follows: no. 4 suo beer (83), No. 3 suo beer (49), No. 12 yangnuo beer (33), No. 7 suo beer (59), No. 5 resident barley (61), No. 7 hua barley (81), No. 1154(6), No. 2 yangnuo beer (5), No. 6 yangnuo beer (42) and No. 2 yunnan beer (85). Wherein Hu 1154 fails to pass the approval and promotion because of its low yield, and the others are barley varieties for beer bred in Jiangsu and other regions. The germplasm has better comprehensive performance and can be directly utilized in production.

Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.

Claims

1. A method for rapidly screening high-quality beer barley germplasm for resisting barley yellow mosaic disease comprises the following steps:

A) the collected and screened n kinds of beer barley varieties (lines) from home and abroad are planted in a barley yellow mosaic disease identification garden for many years, barley yellow mosaic disease resistance investigation is carried out in 2-3 months of the next year, after grains are mature, 6 agronomic characters such as plant height (cm), spike length (cm), single spike grain number (grains), single spike number (grains), single plant grain weight (g) and thousand grain weight (g) are examined, and 1 quality character of a Kuerbaha value (%) is measured.

B) 6 agronomic character performances of the planted n portions of beer barley germplasm are analyzed, and the genetic diversity index of the N portions of beer barley germplasm is researched; 6 agronomic traits and disease levels of the n beer barley germplasms are subjected to correlation analysis, and the correlation among 7 traits is researched. And (3) performing cluster analysis on the germplasm of the n parts of the beer barley planted by taking 8 main traits as indexes. In order to eliminate the influence of correlation among characters, the germplasm evaluation is carried out by adopting principal component analysis. And extracting the first N main components according to the principle that the accumulated contribution rate is greater than 70%.

C) The method comprises the steps of standardizing 8 main character values of the barley germplasm for beer to obtain first N principal component scores, normalizing the N principal component scores, calculating weight coefficients of the principal components according to contribution rates, and comprehensively evaluating various materials by using the sum M of the products of the first N principal component scores and the corresponding weights as a comprehensive index, wherein the higher the M is, the better the comprehensive character performance is, namely the barley germplasm for high-quality barley for beer with resistance to barley yellow mosaic disease can be directly utilized in production.