CN114752593B - SSR (simple sequence repeat) marker for predicting white wax heterosis and application thereof - Google Patents

SSR (simple sequence repeat) marker for predicting white wax heterosis and application thereof Download PDF

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CN114752593B
CN114752593B CN202210365754.1A CN202210365754A CN114752593B CN 114752593 B CN114752593 B CN 114752593B CN 202210365754 A CN202210365754 A CN 202210365754A CN 114752593 B CN114752593 B CN 114752593B
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燕丽萍
吴德军
王因花
任飞
李丽
刘翠兰
贺英俊
张波
张子通
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Shandong Huabo Genetic Engineering Co ltd
Shandong Academy of Forestry
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Abstract

The invention provides an SSR marker for predicting white wax heterosis and application thereof, belonging to the technical field of crop breeding. The SSR molecular marker primer group for predicting the heterosis of the white wax provided by the invention comprises 14 pairs of primers with nucleotide sequences shown as SEQ ID NO.1-SEQ ID NO. 28. According to the genetic distance analysis method based on the molecular level, the hybrid combination of the white wax parents with the genetic distance of 1.1-1.5 is rapidly screened out by adopting SSR markers, parent selection and matching are rapidly obtained, offspring hybrid vigor is obvious, the tree breeding period is shortened by 4-5 years, and the breeding efficiency is remarkably improved.

Description

SSR (simple sequence repeat) marker for predicting white wax heterosis and application thereof
Technical Field
The invention belongs to the technical field of crop breeding, and particularly relates to an SSR marker for predicting white wax heterosis and application thereof.
Background
Heterosis refers to the phenomenon that the first generation of hybrids is superior to the parent in all aspects of body type, growth rate fertility and behavioral characteristics. At present, cross breeding by utilizing heterosis is one of main ways for improving the yield, the resistance and the quality of crops. However, the genetic character of the hybrid F1 generation generated after the hybridization of the parents (male parent and female parent) has uncertainty, not all the hybrid offspring have hybrid advantages, the hybrid advantages of the offspring generated by different parent combinations are quite different, and some hybrid advantages are obvious, some are not obvious, and even degeneration occurs.
One of the most critical factors in utilizing heterosis is the selection of hybrid combinations that give rise to strong vigour. In practical application of woods hybrid vigor breeding, a large number of combinations are prepared and then field phenotype screening is carried out, so that a combined variety with hybrid vigor compared with a control is bred, and the defects of long period, large workload, strong blindness, low efficiency and the like exist, and a plurality of uncertainties of breeding results are caused. And because the tree hybridization breeding period is long and the genetic background is complex, the hybrid parents are blindly selected, so that the utilization of the tree hybridization advantages is lower. The core of cross breeding is the utilization of heterosis, while the selection and matching of parents is a key link in obtaining heterosis. In the past, how to quickly and simply obtain hybrid seeds with new characters or excellent characters meeting production requirements, and how to predict potential heterosis of combined parents in early generation of breeding or in the innovation initial stage of hybrid parents are the most concerned problems in hybridization breeding.
White wax is a generic term for plants of the genus Fraxinus of the family Oleaceae, and is also called Fraxinelly, and is also known as a sand-fixing tree species because of the stocking of Fraxinelly sinensis on the tree. White wax wood is tough and waterproof, and is used in making furniture, farm tools, plywood, etc. and its branches may be basket woven, and the bark is called "cortex Fraxini" and Chinese medicine is used as heat clearing medicine. At present, the prior art does not disclose how to quickly and simply obtain the white wax hybrid with new characters or excellent characters meeting the production requirements.
Disclosure of Invention
Therefore, the invention aims to provide an SSR marker for predicting the hybrid vigor of white wax, which adopts the SSR marker to rapidly screen out the hybrid combination with the genetic distance between parents of 1.1-1.5, rapidly obtain parent selection and match, has obvious offspring hybrid vigor and shortens the tree breeding period by 4-5 years.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides an SSR molecular marker primer group for predicting white wax heterosis, which comprises 14 pairs of primers with nucleotide sequences shown as SEQ ID NO.1-SEQ ID NO. 28.
The invention also provides application of the SSR molecular marker primer group in improved white wax seed selection or parent selection.
The invention also provides application of the SSR molecular marker primer group in the prediction of the hybrid vigor of the white wax.
The invention also provides a method for predicting the heterosis of the white wax, which comprises the following steps: and (3) performing PCR amplification on the parent materials by using the primer group, calculating the genetic distance between the parents by using a Nei genetic distance formula, and screening out hybridization combinations with the genetic distance between the parents being more than 1.
Preferably, the genetic distance is in the range of 1.1-1.5.
Preferably, the white wax includes a downy white wax and a us fraxinus.
Preferably, the heterosis comprises tree height and thoracodiameter.
Preferably, the PCR reaction program is 94 ℃ pre-denaturation for 3min,94 ℃ denaturation for 1min, 54-58 ℃ annealing for 1min,72 ℃ extension for 35 cycles; finally, the reaction was terminated by extension at 72℃for 10min and at 4℃for 10 min.
Preferably, the PCR reaction system comprises the following components in parts by weight: 25mmol/L Mg 2+ 0.8 part, 10 mu mol/L primer 0.2 part, 10mmol/L dNTP 0.3 part, 5U/. Mu.L Taq enzyme 0.05 part, 5-10 ng/. Mu.L DNA template 2.00 parts, 10 XPCR buffer 1.0 part, ddH 2 And 5.45 parts of O.
The invention has the beneficial effects that:
according to the genetic distance analysis method based on the molecular level, the hybrid combination of the white wax parents with the genetic distance of 1.1-1.5 is rapidly screened out by adopting SSR markers, parent selection and matching are rapidly obtained, offspring hybrid vigor is obvious, the tree breeding period is shortened by 4-5 years, and the breeding efficiency is remarkably improved.
Drawings
FIG. 1 is a graph showing the linear correlation of genetic distance with heterosis and mating force.
Detailed Description
The invention provides an SSR molecular marker primer group for predicting white wax heterosis, which comprises 14 pairs of primers with nucleotide sequences shown as SEQ ID NO.1-SEQ ID NO. 28.
In the present invention, the names and specific sequences of the 14 pairs of primer sequences are shown in Table 1:
TABLE 1 SSR molecular marker primer set of the present invention
The invention also provides application of the SSR molecular marker primer group in improved white wax seed selection or parent selection; application of the SSR molecular marker primer group in the prediction of white wax heterosis.
The invention also provides a method for predicting the heterosis of the white wax, which comprises the following steps: and (3) performing PCR amplification on the parent materials by using the primer group, calculating the genetic distance between the parents by using a Nei genetic distance formula, and screening out hybridization combinations with the genetic distance between the parents being more than 1.
In the present invention, the heterosis preferably includes tree height and chest diameter, and the fraxinus chinensis preferably includes fraxinus velutina and fraxinus. The specific calculation method of the Nei genetic distance formula is not particularly limited, and the genetic distance between the parents is calculated through SAS/STAT software. In the present invention, the genetic distance is preferably in the range of 1.1 to 1.5.
The invention researches the relativity of genetic distance between the white wax parents and filial generation heterosis and mating force, adopts 7 white wax parents to carry out incomplete double-row hybridization, configures 10 combinations altogether, calculates general mating force, special mating force and heterosis of the parents, carries out genetic diversity analysis on 7 white wax parent clones by combining SSR markers, and further analyzes the genetic distance of the parentsCorrelation between the true and true vigour of progeny hybrids and the parent mating force. The result shows that the tree height, breast diameter and heterosis of the filial generation of 2 groups of white wax hybrid combinations have larger differences between different hybrid combinations, and reach obvious or extremely obvious levels. The analysis of the coordination force shows that the general coordination force and the special coordination force of the white wax have obvious differences among different characteristics of different parents. The 14 microsatellite markers all show high polymorphism in parents, 49 alleles are detected totally, the polymorphic site percentage is 87.72%, the Nei' S gene effective allele index (Ne) is 2.6656, the Shannon information diversity index (I) is 1.0666, and the genetic diversity level is higher. The Genetic Distance (GD) between different parents is 0.0565-1.7996, and the correlation analysis finds that the genetic distance is equal to the high growth amount of the filial generation tree, the high heterosis of the tree, the general mating force (GCA) of the high father of the tree P2 ) The sum of the general tree-height mating forces and the special tree-height mating force SCA show obvious positive correlation relations, and the correlation coefficients are respectively 0.637, 0.695, 0.700, 0.642 and 0.703. But the correlation with the genetic parameters of the general mating force and chest diameter of the high female parent of the tree is not obvious. The SSR markers of the invention are used for predicting the heterosis of the white wax, and provide reference basis for white wax fine variety breeding and parent selection.
In the invention, when the primer group is used for carrying out PCR amplification on a parent material, the reaction program of the PCR is preferably 94 ℃ pre-denaturation for 3min,94 ℃ denaturation for 1min, 54-58 ℃ annealing for 1min and 72 ℃ extension for 1min, which are 35 cycles in total; finally, the reaction was terminated by extension at 72℃for 10min and at 4℃for 10 min. The reaction system of the PCR is preferably as follows in parts by weight: 25mmol/LMg 2+ 0.8 part, 10 mu mol/L primer 0.2 part, 10mmol/L dNTP 0.3 part, 5U/. Mu.L Taq enzyme 0.05 part, 5-10 ng/. Mu.L DNA template 2.00 parts, 10 XPCR buffer 1.0 part, ddH 2 And 5.45 parts of O. The specific sources of the above components are not particularly limited, and products commercially available in the art can be used.
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
Example 1
In 3 months 2013, 10 groups of hybridization combinations (see Table 1) are obtained by adopting discontinuous factorial mating designs and taking Lu wax No. 3 (Fraxinus velutina 'Lula San hao'), R36-1 (Fraxinus velutina 'R36-1'), R37 (Fraxinus velutina 'R37') as female parents, lu wax No. 4 (Fraxinus velutina 'Lula Si hao'), lu wax No. 5 (Fraxinus pennsylvanica 'Lula Wu hao'), J2 (Fraxinus velutina 'J2') and J5 (Fraxinus velutina 'J5') as male parents, wherein, the Lu Chi 3 (variety weight number 20100013) and the Lu Chi 4 (variety weight number 20100014) are new varieties of salt-tolerant Fraxinus velutina (Fraxinus velutina) selected by Shandong province forestry science institute, and the Lu Chi 5 (variety weight number 20100015) is a new variety of fast-growing American red fraxinus (Fraxinus pennsylvanica) selected by Shandong province forestry science institute; r36-1, R37, J2 and J5 are excellent Fraxinus velutina novel germplasm collected and bred in the research.
Harvesting hybrid seeds in 11 middle of 2013, and simultaneously collecting half-sibling family seeds of 3 female parents as a control. Seedling raising is carried out in a test nursery of Shandong forestry science institute at the end of 3 months in 2014. Soaking the hybrid seeds in water at 35-40 deg.c for 5-7 d, and changing water once every other day. Sowing and seedling raising are carried out by adopting a light matrix non-woven fabric container, wherein the diameter of the container is 12cm, and the height of the container is 15cm. The matrix comprises grass carbon: vermiculite V/v=3: 1, the substrate was sterilized with 0.15% carbendazim. The substrate in the container is irrigated thoroughly one day before sowing, 2-3 seeds are sowed in each container, holes are dug in advance, the depth of each hole is 1-1.5 cm, vermiculite with the thickness of 1-2 cm is used for covering after sowing, and watering is carried out immediately.
4 months 2015, the hybrid seedlings bred by the light matrix non-woven fabric container are subjected to complete random block test design, 10 plants are planted in each cell, the line spacing is 3m multiplied by 4m, the hybrid seedlings are planted in a life light test base of Shandong forestry science research institute, water is poured in time, soil is covered after water permeates, and conventional tending management is adopted in test forests. The test forest land is located at the east longitude of 118 degrees 32 '-119 degrees 10', the north latitude of 36 degrees 41 '-37 degrees 19', the annual average air temperature of 13.2 ℃, the annual average precipitation of 708.4 mm, the standing condition and the like. After the tree leaves fall at the bottom of 10 months in 2019, the tree height and breast diameter of the half sibling family progeny of the 5-year 10 hybrid combined progeny and 3 parents are investigated plant by plant.
Hybrid vigor calculating highwayFormula (Li Yiliang, etc., 2012) is: h= (F) i -MP)/MP×100%, where F i The average value of the characteristics of any combined hybrid offspring is obtained, and MP is the average value of the characteristics of the free pollination offspring of each parent. The analysis methods of parameters such as linear model of each trait analysis of variance, analysis of mating force analysis of variance and genetic force are referred to (Dong Hong, etc., 2017). SAS/STAT software was used for each trait analysis of variance and for the force of variance analysis.
The tree height and breast diameter of 5-year-old test forest at the longevity test station are investigated in order to select hybrid combinations excellent in comprehensive properties. The height and the chest diameter of a 5-year tree of a full-sibling family of 10 hybrid combined filial generations and a female parent R37, R36 and R36-1 half-sibling family are shown in Table 1, the height of a No.2 hybrid combination R37 XLu wax No. 5 tree between the Fraxinus velutina and the Fraxinus velutina is 6.42m at the highest, a No.1 hybrid combination R37 XLu wax No. 4 in the Fraxinus velutina is 5.93m, and the minimum combination is a No. 16 Fraxinus velutina inner combination R36-1 XLu wax No. 4 is 4.68m; the maximum difference between the trees Gao Bianfu, 42m and 4.68m is 1.74m, which shows that the tree height variation between different hybridization combinations is larger. The maximum combination of the chest diameters is the number 2 between seeds, the minimum combination is the number 7 in seeds, the amplitude is 8.98 cm-6.51 cm, and the maximum difference is 2.47cm. The standard deviation of the height of each hybrid combined tree is 0.01 m-0.59 m, the variation coefficient is 0.20-11.72%, the standard deviation of the breast diameter is 0.03 cm-0.97 cm, and the variation coefficient is 0.42-13.59%. Thus, there is a wide variation in progeny growth traits among the various hybrid combinations, and it is feasible to select superior hybrid combinations.
TABLE 1 analysis of filial generation traits
Whether these variations among the filial generations of the hybrid combination reach significant levels or not needs to be verified by analysis of variance, and the analysis of variance results of the individual traits in table 1 are shown in table 2. As can be seen from the table, the differences between the hybridization combinations of the tree height and breast diameter traits all reach an extremely significant level (P < 0.01).
TABLE 2 analysis of variance for individual traits of hybrid combinations
From the above analysis, there was a wide variation in the height and diameter of the chest between the hybrid combinations and very significant levels were reached. Whether these variations exist between hybridization combinations due to genetic or environmental factors, needs to be demonstrated by the parameter genetic strength. Genetic transmission refers to the ratio of genetic variance to phenotypic variance. The genetic control of the character is strong, and the influence of the environment is weak. As is clear from Table 2, the genetic power of the tree height and chest diameter traits was all greater than 0.7 or more. Therefore, the growth characteristics of the white wax hybrid progeny are mainly controlled by genetic factors and less affected by the environment.
The results of multiple comparison and heterosis analysis of the traits are shown in Table 3, the tree height, chest diameter and other hybridization combinations of the hybridization combinations of No.2 reach extremely significant differences of 1%; the height and the chest diameter of the 6 # hybrid combination reach 5% of the obvious difference with other hybrid combinations; the height of the No.1 hybrid combination reaches 5% of the significant difference with other hybrid combinations, and the difference of the chest diameters is not significant. Indicating that there is a significant difference in growth traits between the progeny of each hybrid combination, it is feasible to select superior hybrid combinations to improve hybrid vigor utilization.
From the results of heterosis in Table 3, it can be seen that there is a large difference in heterosis between the different combinations. The hybridization advantages of the height and the diameter of the tree in the Fraxinus velutina seed are varied from-6.92 to 15.37 percent and-7.26 to 7.43 percent. The hybrid vigor of the height and chest diameter of the fraxinus velutina and the fraxinus velutina is changed to-1.25% -24.9% and-5.84% -25.77%, and the hybrid vigor between the seeds is superior to that of the seed. The hybrid vigor of the hybrid combination tree with the height and the chest diameter of the No.2 is highest, and the No. 6 shows that the 2 groups of hybrid combinations can create basic groups with rich variation for the breeding of the next generation through the hybridization among parents, and the hybrid offspring is easy to obtain higher hybrid vigor.
TABLE 3 hybrid seedling growth trait manifestation and heterosis
Note that: the spss test shows that the difference is significant (P < 0.05) in the lower case letters of the same column.
The results of the growth trait mating force analysis are shown in Table 4. The general coordination force effect values of the white wax have obvious differences among different characters of different parents, and the general coordination force effect values of the high 7 parent trees are that Lu wax No. 5 is more than R37 = Lu wax No. 3 is more than J2 is more than Lu wax No. 4 is more than J5 is more than R36-1 in sequence; the general coordination force effect values of the chest diameters of the 7 parents are that Lu wax No. 5 > R37 > Lu wax No. 3> J2 > Lu wax No. 4 > R36-1 > J5 in sequence. The genetic characteristics of the parents are different, wherein the tree height and the chest diameter general mating force effect value of the female parent R37 and the male parent Lu-wax No. 5 are both larger, so that the female parent R37, the Lu-wax No. 3 and the male parent Lu-wax No. 5 are the parents with higher breeding values, and the hybrid combination of the parents is easier to obtain excellent filial generation as the preferential parents in future hybrid breeding.
Analysis of the special mating force effect of the tree height and the ground diameter of 10 hybrid combined whole-sibling families shows that the results in table 4 show that the differences of the various characters of the tree height and the breast diameter of different hybrid combined families are larger, and the special mating force effect value amplitude of the tree height and the breast diameter is respectively-0.66 to 1.04 and-0.42 to 0.95. The R37 XLu wax No. 5 combined family has maximum special coordination force effect values of 1.04 and 0.95 on the tree height and the ground diameter, and the R37 XLu wax No. 3 XLu wax No. 5 combined family has special coordination force effect values of 0.79 and 0.80 on the tree height and the ground diameter, so that the higher the special coordination force effect value is, the more obvious the heterosis is, and therefore the hybridization of R37 XLu wax No. 5 and Lu wax No. 3 XLu wax No. 5 can be used as the preferable combination of white wax hybridization breeding.
TABLE 4 hybrid seedling trait analysis
Example 2
The 7 parent materials described in example 1 were subjected to PCR amplification using 14 pairs of primer sequences shown in Table 5, specifically: collecting current-year tender leaves of the 7 parents, extracting DNA and carrying out SSR marker test. The genome DNA is extracted by adopting a CTAB method, and the genome DNA is slightly improved, and 2 percent of beta-mercaptoethanol and 2 percent of PVP are added into an extraction buffer solution. The concentration and purity of the DNA were determined by UV spectrophotometry and diluted to 30 ng/. Mu.l.
SSR-PCR employed 10. Mu.l reaction system: 25mmol/l Mg 2+ 0.8. Mu.l, 10. Mu. Mol/l primer 0.2. Mu.l, 10mmol/l dNTP 0.3. Mu.l, 5U/. Mu.l Taq enzyme 0.05. Mu.l, 5-10 ng/. Mu.l DNA template 2.00. Mu.l, 10 XPCR buffer 1.0. Mu.l, ddH 2 O5.45. Mu.l; the PCR reaction conditions are 94 ℃ pre-denaturation for 3min,94 ℃ denaturation for 1min, 54-58 ℃ annealing for 1min,72 ℃ extension for 1min, and 35 cycles are total; finally, the reaction was terminated by extension at 72℃for 10min and at 4℃for 10 min.
And (3) polyacrylamide gel electrophoresis detection: the PCR products were detected by 6% denaturing PAGE electrophoresis, and after the electrophoresis was completed, the results were observed by staining with silver nitrate. The amplified bands were read according to the PAGE gel electrophoresis results, each SSR amplification site was regarded as 1 gene site, each site was identified as AA, BB, CC, AB, BC, AC, and so on, constituting the original data. Using the obtained SSR data, the polymorphic site percentage (PPL), shannon information diversity index (I), nei's gene diversity index (HE) was calculated using popkene software.
As a result, as shown in Table 5, the number of amplified loci per pair of primers was varied from 76 to 187, the allele factors per locus were 2 to 4, 49 alleles were detected in total, and the average allele factor per locus was 3.5. The total number of amplified bands of 14 pairs of SSR primers in the parent population is 57, the average number of amplified bands of each pair of primers is 4.07, the number of polymorphic bands is 50, the polymorphic site percentage (PPL) is 87.72%, the Nei' S gene effective allele index (Ne) is 2.6656, and the Shannon information diversity index (I) is 1.0666.
TABLE 5 SSR primer amplification number of sites and polymorphism information content
Genetic distances (Dg) between parents were calculated by SAS/STAT software using the Nei genetic distance formula, and the results are shown in table 6: the genetic distance amplitude between 6 parents (R37, lu wax No. 3, R36-1, lu wax No. 4, J5 and J2) in the Fraxinus velutina is 0.0565-0.9444, and the average genetic distance is 0.6472; the genetic distance between the Fraxinus velutina parent (R37, fraxinus velutina No. 3 and R36-1) and the Fraxinus velutina parent (Fraxinus velutina No. 5) is 1.1985-1.7996, the average genetic distance is 1.4753, and the coefficient of variation is 40.74%. The genetic distance between the Fraxinus velutina and the parents of the Fraxinus velutina species is far greater than the genetic distance between the parents of the Fraxinus velutina species. The genetic distances among the parents of the 10 hybridization combinations are greatly different, the maximum genetic distance between R36-1 and Lu wax No. 5 is 1.7996, and the minimum genetic distance between R36-1 and Lu wax No. 4 is 0.0565. Wherein the parental genetic distance between the R37×Lu wax No. 5 combination and the Lu wax No. 3×Lu wax No. 5 combination with obvious heterosis is 1.1985-1.4277.
TABLE 6 genetic distance between parents
Example 3
And (5) correlation analysis of heterosis and parent genetic distance. And constructing a scatter diagram of the variation of the offspring heterosis along with the parent mating force and the genetic distance in Microsoft Office Excel, detecting the fitting degree of the offspring heterosis, the parent mating force and the genetic distance unitary linear regression model through software, and establishing a corresponding unitary linear regression equation. Genetic distances among parents are respectively related to offspring heterosis and coordination force by using spps 21.0 statistical analysis software, and the results are shown in table 7.
The SSR marker genetic distance and the high growth amount of the filial generation tree, the high heterosis of the tree, the general mating force of the high male parent and the general mating force of the tree are all in obvious positive correlation, the correlation coefficient is between 0.642 and 0.703, and the correlation with the general mating force of the high female parent is not obvious. SSR marker genetic distance and chest diameterThe correlation analysis of the genetic parameters of the SSR marker shows that the correlation is not obvious, the comparison of the correlation coefficients shows that the SSR marker genetic distance is highest in correlation coefficient with the special mating force, and the male parent is inferior in general mating force. Comparing the SSR mark genetic distance with the correlation coefficient of tree height and breast diameter, finding that the correlation coefficient of the SSR mark genetic distance with the general mating force and the special mating force of the male parent is the highest, and the correlation coefficient is between 0.544 and 0.703; but the correlation coefficient of the general coordination force with the female parent is lower than 0.241-0.244. Further by regression analysis it was found that: genetic distance between parents, tree vigour, chest diameter vigour, parent GCA P1+P2 And SCA (Table 7). The significance test result shows that the parent genetic distance and the tree vigour are high, and the parent GCA P1+P2 And SCA reach significant or very significant levels, while none of the other linear equation tests reach significant or very significant levels.
TABLE 7 correlation of white wax genetic distance with heterosis and coordination force
The correlation of SSR genetic distances between parents and progeny growth traits can be more intuitively reflected in the scatter plot of parent genetic distances and progeny growth traits (fig. 1). With the increase of the genetic distance between parents, the tree has high hybrid vigour and the parents GCA P1+P2 And SCA are significantly increased, tending to correlate positively when the genetic distance is greater than 1. The scatter diagram shows that the correlation coefficient of the parent genetic distance and the tree height special mating force is 0.4936 at the maximum, and the correlation coefficient of the parent genetic distance and the tree height male parent general mating force GCA is 0.4899, and the scatter diagram is matched with the regression line comparison analysis. Further by regression analysis it was found that: genetic distance between parents, tree vigour, chest diameter vigour, parent GCA P1+P2 And SCA (Table 7). The significance test result shows that the parent genetic distance and the tree vigour are high, and the parent GCA P1+P2 And SCA reach significant or very significant levels, while none of the other linear equation tests reach significant or very significant levels.
The female parent 'R37', 'Lu wax 3' and the male parent 'Lu wax 5' can be obtained through the above embodiment, generally the combination force is higher, wherein the R37 XLu wax 5 and the Lu wax 3 XLu wax 5 combination families have the maximum special combination force effect value on the tree height and the ground diameter, and the combination hybridization advantage is also strongest. The higher the general fitness of the parent, the higher the specific coordination effect value, thus indicating that the general fitness of the parent is one of the important factors affecting the vigour of the combined hybrid. Therefore, the parents with higher general combining force are firstly selected for matching, and then the combination with high special combining force is selected, so that the hybrid vigor of the combination can be effectively improved, and the hybrid combination with strong vigor is screened.
The matching force has stronger prediction capability on the growth character expression and the heterosis of the offspring, and can be used as an important basis for selecting parent and predicting the heterosis of the offspring. The linear correlation result of genetic distance and heterosis and coordination force shows that GD (genetic distance) and offspring growth character phenotype value, heterosis and parent GCA P1+P2 (sum of general binding forces of parents), GCA P2 (father general mating force), SCA (Special mating force) show a significant positive correlation, GD and GCA P1+P2 The offspring phenotype value and heterosis have the strongest correlation, and GCA P2 Next, GCA P1 At the weakest, GCA is used for selective breeding P1+P2 Prediction of sub-representative values and heterosis should be compared to GCA P1 And GCA P2 The accuracy is higher. Therefore, in breeding, GCA can be preferentially adopted for parent selection of established forests P1+P2 Predicting the phenotype value of the growth trait of the offspring, and predicting the heterosis of the growth trait of the offspring by adopting SCA; and for parent selection of the non-constructed forest, the prediction of the offspring growth character phenotype value and the hybrid vigor can be realized more accurately based on genetic distance analysis of a certain level.
The result of the embodiment of the invention shows that as the genetic distance between parents increases, the tree height and thoracodiameter hybrid vigor both increase, and when the genetic distance is greater than 1, the forward correlation relation tends to be towards, and the vigor of the hybrid vigor with the tree height increases remarkably. I.e. the greater the probability that a hybrid combination with a distant genetic distance of the parents will result in a strong hybrid vigour. The practical possibility of carrying out the analysis of white wax genetic diversity, the prediction of heterosis and the like by adopting SSR molecular markers is shown. However, the correlation coefficient between the genetic distance and the heterosis obtained in the study is 0.695, and the comparison study of 10 combined heterosis and the genetic distance shows that the larger the genetic distance is, the stronger the heterosis effect is, but the 2 hybrid combinations with the genetic distance ranging from 1.1985 to 1.4277 show strong advantages. Therefore, the genetic distance is assembled among parents within a certain range, which is more beneficial to the expression of heterosis. Therefore, the parent with the genetic distance between the female parent and the male parent being more than 1 is screened, and obvious heterosis can be obtained. Particularly, the invention carries out genetic distance analysis based on molecular level, adopts SSR markers to rapidly screen out hybridization combinations with genetic distances of 1.1-1.5 between parents, can obtain parent selection without constructing offspring to measure forests, has obvious offspring heterosis, can greatly shorten the forest breeding period, and shortens the forest breeding period by 4-5 years.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Sequence listing
<110> Shandong province forestry science institute
SHANDONG HUABO GENETIC ENGINEERING Co.,Ltd.
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Claims (9)

1. An SSR molecular marker primer group for predicting white wax heterosis is characterized by comprising 14 pairs of primers with nucleotide sequences shown in SEQ ID NO.1-SEQ ID NO. 28.
2. The application of the SSR molecular marker primer set in the improved white wax seed selection or parent selection as claimed in claim 1.
3. The use of the SSR molecular marker primer set of claim 1 in fraxinus chinensis hybrid vigour prediction.
4. A method for predicting white wax heterosis comprising the steps of: PCR amplification is performed on parent materials by using the primer set as set forth in claim 1, genetic distances among the parents are calculated by using a Nei genetic distance formula, and hybridization combinations with genetic distances among the parents being more than 1 are screened.
5. The method of claim 4, wherein the genetic distance is in the range of 1.1-1.5.
6. The method of claim 4 wherein said white wax comprises a downy white wax and a fraxinello.
7. The method of claim 4, wherein the heterosis comprises tree height and thoracodiameter.
8. The method of claim 4, wherein the PCR is performed at 94℃for 3min,94℃for 1min,54℃to 58℃for 1min, and 72℃for a total of 35 cycles; finally, the reaction was terminated by extension at 72℃for 10min and at 4℃for 10 min.
9. The method according to claim 4, wherein the PCR reaction system comprises the following components in parts by weight: 25mmol/LMg 2+ 0.8 part, 10 mu mol/L primer 0.2 part, 10mmol/L dNTP 0.3 part, 5U/. Mu.LTaq enzyme 0.05 part, 5-10 ng/. Mu.L DNA template 2.00 parts, 10 XPCR buffer 1.0 part, ddH 2 And 5.45 parts of O.
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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104046697A (en) * 2014-07-04 2014-09-17 山东省林业科学研究院 SSR (Simple Sequence repeats) primer group based on Fraxinus Velutina Torr. transcriptome sequencing information development and application of primer group in germplasm identification
CN105861643A (en) * 2015-12-03 2016-08-17 山东农业工程学院 Construction method for fraxinus velutina SSR marked molecular genetic linkage map
CN106244700A (en) * 2016-08-24 2016-12-21 山东省林业科学研究院 A kind of method utilizing SSR marker technology that Fraxinus velutina and American Red cenospecies are carried out Rapid identification
CN109652411A (en) * 2018-12-28 2019-04-19 山东省林业科学研究院 Fluorescent SSR primer combination and application thereof in construction of molecular fingerprint of new variety of white wax

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Publication number Priority date Publication date Assignee Title
CN104046697A (en) * 2014-07-04 2014-09-17 山东省林业科学研究院 SSR (Simple Sequence repeats) primer group based on Fraxinus Velutina Torr. transcriptome sequencing information development and application of primer group in germplasm identification
CN105861643A (en) * 2015-12-03 2016-08-17 山东农业工程学院 Construction method for fraxinus velutina SSR marked molecular genetic linkage map
CN106244700A (en) * 2016-08-24 2016-12-21 山东省林业科学研究院 A kind of method utilizing SSR marker technology that Fraxinus velutina and American Red cenospecies are carried out Rapid identification
CN109652411A (en) * 2018-12-28 2019-04-19 山东省林业科学研究院 Fluorescent SSR primer combination and application thereof in construction of molecular fingerprint of new variety of white wax

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中国引种桉树种质资源的遗传多样性分析;刘果;谢耀坚;陈鸿鹏;吴志华;;西北植物学报(第11期);全文 *

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