CN114752593A - SSR marker for predicting white wax heterosis and application thereof - Google Patents
SSR marker for predicting white wax heterosis and application thereof Download PDFInfo
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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 white wax heterosis provided by the invention comprises 14 pairs of primers with nucleotide sequences shown as SEQ ID NO.1-SEQ ID NO. 28. The invention is based on a molecular level genetic distance analysis method, adopts SSR markers to quickly screen out the hybrid combination with the genetic distance between the fraxinus chinensis parents of 1.1-1.5, quickly obtains the parent matching, has remarkable progeny hybrid vigor, shortens the forest breeding period by 4-5 years, and obviously improves the breeding efficiency.
Description
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 is a phenomenon that the first generation of hybrids is superior to the parents in body type, growth rate, fertility and behavior characteristics. At present, crossbreeding by utilizing heterosis is one of the main ways for improving the yield, resistance and quality of crops. However, genetic traits of the F1 hybrid generation generated after hybridization of parents (male parent and female parent) are uncertain, not all filial generations have hybrid vigor, the hybrid vigor of the progeny generated by different parent combinations is also different, some hybrid vigors are obvious and some are not obvious, and even degeneration occurs.
One of the most critical factors in utilizing heterosis is the selection of a strong heterosis hybrid combination. In the practical application of forest heterosis breeding, a large number of combinations are prepared and then subjected to field phenotype screening, so that a combined variety with heterosis compared with a contrast is bred, and the defects of long period, large workload, strong blindness, low efficiency and the like exist, so that various uncertainties of breeding results are caused. Moreover, as the tree crossbreeding period is long, the genetic background is complex, and the crossbreeding parents are selected blindly, the utilization of the crossbreeding advantages of the tree breeding is low. The core of hybrid breeding is to utilize heterosis, and the selection and matching of parents are key links for obtaining the heterosis. Conventionally, it has been a problem of great concern in hybrid breeding that how to obtain a hybrid having a new trait or an excellent trait satisfying production requirements quickly and simply and how to predict the potential heterosis of a combination parent in an early generation of breeding or in an early stage of innovation of the hybrid parent.
Fraxinus chinensis is a generic name of plant of Fraxinus of Oleaceae, also called Fraxinus negra, and is called Fraxinus chinensis, which is also called as species of Fraxinus rhynchophylla. The white wax wood is tough and waterproof, can be used for manufacturing furniture, farm implements, plywood and the like, branches can be woven into baskets, bark is called ash bark, and the white wax wood is used as a heat clearing medicine in traditional Chinese medicine. At present, the prior art does not disclose how to rapidly and simply obtain a white wax hybrid having a new or excellent trait meeting the production requirements.
Disclosure of Invention
In view of the above, the invention aims to provide an SSR marker for predicting the white wax heterosis, wherein a hybrid combination with genetic distance between parents of 1.1-1.5 is rapidly screened by the SSR marker, the parents are rapidly obtained for matching, the heterosis of progeny is remarkable, and the forest breeding period is shortened 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 white wax fine variety breeding or parent selection.
The invention also provides application of the SSR molecular marker primer group in the prediction of the white wax heterosis.
The invention also provides a method for predicting the white wax heterosis, which comprises the following steps: and carrying out 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 the hybridization combination with the genetic distance between the parents being more than 1.
Preferably, the genetic distance is in the range of 1.1 to 1.5.
Preferably, the white wax comprises fluffy white wax and Fraxinus americana.
Preferably, the heterosis includes tree height and breast diameter.
Preferably, the reaction program of the PCR is pre-denaturation at 94 ℃ for 3min, denaturation at 94 ℃ for 1min, annealing at 54-58 ℃ for 1min, and extending lmin at 72 ℃, and the reaction program comprises 35 cycles; finally, extension is carried out at 72 ℃ for 10min, and the reaction is terminated at 4 ℃ for 10 min.
Preferably, the reaction system of the PCR comprises the following components in parts by weight: 25mmol/L Mg2+0.8 parts, 0.2 parts of 10 mu mol/L primer, 0.3 parts of 10mmol/L dNTP, 0.05 parts of 5U/mu L Taq enzyme, 2.00 parts of 5-10 ng/mu L DNA template, 1.0 parts of 10 XPCR buffer solution, ddH2And 5.45 parts of O.
The invention has the beneficial effects that:
the invention is based on a genetic distance analysis method of molecular level, adopts SSR markers to quickly screen out the hybrid combination with the genetic distance between fraxinus chinensis parents of 1.1-1.5, quickly obtains parent matching, has remarkable progeny hybrid vigor, shortens the forest breeding period by 4-5 years, and obviously improves the breeding efficiency.
Drawings
FIG. 1 is a graph showing a linear correlation between genetic distance and heterosis and combining ability.
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 invention
The invention also provides application of the SSR molecular marker primer group in fine variety breeding or parent selection and matching of Fraxinus chinensis; and the application of the SSR molecular marker primer group in the prediction of the heterosis of the white wax.
The invention also provides a method for predicting the white wax heterosis, which comprises the following steps: and carrying out 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 the hybridization combination with the genetic distance between the parents being more than 1.
In the present invention, the heterosis preferably includes tree height and breast diameter, and the fraxinus comprises fraxinus tomentosa and fraxinus spicata. The specific calculation method of the Nei genetic distance formula is not specially limited, and the genetic distance between 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 correlation between the genetic distance between the white wax parents and the heterosis and the combining ability of the filial generation, specifically adopts 7 white wax parents to carry out incomplete double-row hybridization, totally configures 10 combinations, calculates the general combining ability, the special combining ability and the heterosis of the parents, and combines SSR marks to carry out genetic diversity analysis on the clone of the 7 white wax parents, thereby analyzing the correlation between the genetic distance of the parents and the heterosis and the combining ability of the filial generation. The results show that the phenotypic values of the tree height and the breast diameter of the filial generation of the 2 groups of fraxinus chinensis hybrid combinations and the heterosis have larger differences among different hybrid combinations and reach obvious or extremely obvious levels. The combining ability analysis finds that the general combining ability and the special combining ability of the white wax have obvious difference between different properties of different parents. The 14 microsatellite markers are highly polymorphic in parents, 49 alleles are detected in total, the percentage of polymorphic sites is 87.72 percent, the effective allele index (Ne) of the Nei' S gene 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 correlation analysis shows that the genetic distance and the hybrid progeny tree have high growth quantity, high hybrid vigor and high male parent General Combining Ability (GCA) P2) The sum of general combining ability of the tree heights and the special combining ability SCA of the tree heights are in a positive correlation, and the correlation coefficients are 0.637, 0.695, 0.700, 0.642 and 0.703 respectively. The correlation between the general combining ability of the high female parent of the tree and the genetic parameters of the diameter at breast height is not obvious. The SSR marker is feasible to predict the hybrid vigor of the fraxinus chinensis, and provides reference basis for fine variety breeding and parent selection of the fraxinus chinensis.
In the present invention, when the above primer set is used to perform PCR amplification on the parent material, the reaction procedure of the PCR is preferably pre-denaturation at 94 ℃ for 3min and denaturation at 94 ℃ for 1min, annealing at 54-58 ℃ for 1min, extending at 72 ℃ for l min, and 35 cycles; finally, extension is carried out at 72 ℃ for 10min, and the reaction is terminated at 4 ℃ for 10 min. The reaction system of the PCR is preferably as follows according to the parts by weight: 25mmol/LMg2+0.8 part, 0.2 part of 10 mu mol/L primer, 0.3 part of 10mmol/L dNTP, 0.05 part of 5U/mu L Taq enzyme, 2.00 parts of 5-10 ng/mu L DNA template, 1.0 part of 10 XPCR buffer solution, ddH2And 5.45 parts of O. The specific sources of the above components are not particularly limited in the present invention, and any commercially available product that is conventional in the art may 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 of 2013, selecting Shandong province forest industry No. 3 (Fraxinus velutina ' Lu la San hao '), R36-1(Fraxinus velutina ' R36-1 '), R37(Fraxinus velutina ' R37 ') as female parent, Shandong wax No. 4 (Fraxinus velutina ' Lu la Si hao '), Shandong wax No. 5(Fraxinus pennsylvania ' Lu la Wu hao '), J2(Fraxinus velutina ' J2 ') and J5(Fraxinus velutina ' J5 ') as male parent, and adopting discontinuous mating to obtain 10 groups of hybrid combinations (shown in Table 1), wherein Shandong wax No. 3 (variety No. 20100013) and Shandong province forest industry No. 4 (Fraxinus velutina ' Lu J49376) are new varieties of Shandong forest industry research, and the Shandong province forest industry No. 5 is new variety of Shandong forest industry No. 5 scientific Shandong forest industry No. 5(Fraxinus sanyu Shandong forest industry No. 5) as new variety research; r36-1, R37, J2 and J5 are new excellent Fraxinus velutina germplasm collected and bred in the research.
In 11 th month in 2013, hybrid seeds are collected, and meanwhile, semi-sib family seeds of 3 female parents are collected as a reference. The 3 th month in 2014 is at the end of the trial of the forest industry scientific research institute in Shandong province for nursery seedling culture. The hybrid seeds are soaked in water of 35-40 ℃ for 5-7 days, and the water is changed every other day. Seeding and seedling raising are carried out by adopting a light-matrix non-woven fabric container, and the diameter of the container is 12cm, and the height of the container is 15 cm. The matrix comprises the following components: vermiculite V/V is 3: 1, sterilizing the substrate with 0.15% carbendazim. And (3) thoroughly watering the substrate in the container one day before sowing, sowing 2-3 seeds in each container, digging holes in advance with the hole depth of 1-1.5 cm, covering the seeds with vermiculite with the thickness of 1-2 cm after sowing, and immediately watering.
In 2015 for 4 months, hybrid seed seedlings bred in a light-matrix non-woven fabric container are subjected to complete random block test design, 10 plants are planted in each cell for 4 times, the plant spacing and the row spacing are 3m multiplied by 4m, the hybrid seed seedlings are planted in a shogay test base of Shandong province forestry science research institute, water is timely irrigated to penetrate water, soil is covered after water penetrates, and test forests are subjected to conventional tending management. The test forest land is at 118 degrees 32 'to 119 degrees 10' of east longitude, 36 degrees 41 'to 37 degrees 19' of north latitude, 13.2 ℃ of annual average temperature, 708.4 mm of annual average precipitation, and medium conditions of land standing. After the tree leaves fall in 2019 in the bottom of 10 months, the tree height and the breast height are investigated one by one for 10 hybrid combined filial generations in 5 years and half-sib family filial generations of 3 parents.
The heterosis calculation formula (Liyiliang et al, 2012) is: h ═ Fi-MP)/MP X100%, wherein, FiThe average value of the characters of any one combined hybrid filial generation, and the MP is the average value of the characters of the freely-pollinated filial generation of each parent. Parametric analysis methods such as an analysis of variance for each property linear model, analysis of variance for combining ability, and heritability are referred to (report on the subject, etc., 2017). SAS/STAT software is adopted for analysis of variance of each character and analysis of variance of combining ability.
The tree height and breast diameter of the 5-year test stand of the longevity light test stand are investigated, so as to select a hybrid combination with excellent comprehensive character performance. The height and diameter at breast height of 5-year trees of 10 hybrid combination progeny hologynes and female parents R37, R36 and R36-1 half-sib families are shown in Table 1, the height of the No. 2 hybrid combination R37 XU wax No. 5 trees in Fraxinus rhynchophylla is 6.42m at most, the height of the No. 1 hybrid combination R37 XU wax No. 4 in Fraxinus rhynchophylla is 5.93m, and the minimum combination is the No. 16 internal hybrid combination R36-1 XU wax No. 4 in Fraxinus rhynchophylla is 4.68 m; the tree height variation is 6.42 m-4.68 m, and the maximum difference is 1.74m, which indicates that the tree height variation among different hybridization combinations is large. The maximum combination of the breast diameters is No. 2 between seeds, the minimum combination is No. 7 in the seeds, the amplitude is 8.98 cm-6.51 cm, and the maximum difference is 2.47 cm. The standard deviation of the height of each hybridization combination tree is 0.01-0.59 m, the coefficient of variation is 0.20-11.72%, the standard deviation of the breast diameter is 0.03-0.97 cm, and the coefficient of variation is 0.42-13.59%. Therefore, there is wide variation in the progeny growth traits among the respective hybridization combinations, and it is feasible to select a good hybridization combination.
TABLE 1 analysis of hybrid progeny traits
Whether the variation existing among filial generation of the hybrid combination reaches a remarkable level needs to be verified by anova, and the anova result of each character in the table 1 is shown in a table 2. As can be seen from the table, the differences between the hybridization combinations of the tree height and breast diameter traits reach a very significant level (P < 0.01).
TABLE 2 analysis of variance of individual traits for hybridization combinations
From the above analysis, it can be seen that there is wide variation in tree height and breast diameter between hybridization combinations, and it reaches a very significant level. Whether these variations between hybridization combinations are caused by genetic or environmental factors requires the demonstration of the heritability parameter. Heritability refers to the ratio of genetic variance to phenotypic variance. The heritability of the character is large, which shows that the character is strongly controlled by heredity and is weakly influenced by environment. As can be seen from Table 2, the heritability of the tree height and diameter at breast height is greater than 0.7. Therefore, the growth traits of the fraxinus chinensis hybrid progeny are mainly controlled by genetic factors and are less influenced by the environment.
The results of multiple comparison and heterosis analysis of various characters are shown in table 3, the tree height and breast diameter of No. 2 hybrid combination are very different from those of other hybrid combinations by 1%; the tree height and breast diameter of the No. 6 hybrid combination are all 5% different from those of other hybrid combinations; the tree height of the hybridization combination No. 1 and other hybridization combinations reach 5 percent of obvious difference, but the chest diameter difference is not obvious. The obvious difference of growth performance among filial generations of each hybridization combination is shown, and the selection of excellent hybridization combination to improve the utilization of heterosis is feasible.
As can be seen from the heterosis results in Table 3, the heterosis differences between the different combinations are large. The hybrid dominance change of the internal tree height and the breast diameter of the Fraxinus velutina is-6.92% -15.37% and-7.26% -7.43%. The hybrid vigor of the height and the diameter at breast height between species of Fraxinus velutina and Fraxinus rhynchophylla is changed from-1.25 to 24.9 percent and from-5.84 to 25.77 percent, and the hybrid vigor between species is superior to the hybrid vigor in species. The hybrid vigor of the No. 2 hybrid combination tree is the highest and the hybrid vigor of the breast diameter is the next No. 6, which shows that the 2 hybrid combinations can create basic groups with rich variation for the breeding of the next generation through the hybridization between parents, and the filial generation can easily obtain higher hybrid vigor.
TABLE 3 expression of growth traits of hybrid seedlings and heterosis
Note: the different lowercase letters in the same column indicate significant difference (P < 0.05) by the sps method test.
The results of the growth trait binding capacity analysis are shown in Table 4. The general combining ability effect value of the white wax is obviously different among different parent different properties, and the general combining ability effect value with the height of 7 parent trees is 5 # R37 ═ Rouzu 3 # J2 # Rouzu 4 # J5 # R36-1 in sequence; the general combining force effect values of 7 parent breast diameters are 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 chest diameter general combining ability effect values of the female parent R37 and the male parent Lu wax No. 5 are larger, so that the female parent R37, the Lu wax No. 3 and the male parent Lu wax No. 5 are parents with higher breeding values, and are used as preferred parents in future crossbreeding, and excellent filial generation can be obtained more easily by the crossbreeding combination.
The special combining ability effect of 10 hybridization combination holomorphic family trees in height and ground diameter is analyzed, and the results in Table 4 show that the differences of the tree heights and the breast diameters of different hybridization combination families in all characters are large, and the special combining ability effect values of the tree heights and the breast diameters are respectively-0.66-1.04 and-0.42-0.95. The R37 XLu wax 5 combined family line has the maximum special combining force effect values of 1.04 and 0.95 on the height and the ground diameter of a tree, and the second Lu wax 3 XLu wax 5 combined family line has the special combining force effect values of 0.79 and 0.80 on the height and the ground diameter of a tree, the higher the special combining force effect value is, the more obvious the heterosis is, so the R37 XLu wax 5 and Lu wax 3 XLu wax 5 hybrid can be used as the preferable combination for white wax hybrid breeding.
TABLE 4 analysis of hybrid trait
Example 2
The PCR amplification was performed on 7 parent materials described in example 1 using 14 primer sequences shown in Table 5, specifically: collecting the current year tender leaves of the 7 parents, extracting DNA and carrying out SSR marker test. The CTAB method is adopted to extract genome DNA, and is slightly improved, 2 percent of beta-mercaptoethanol and 2 percent of PVP are added into an extraction buffer solution. The concentration and purity of the DNA was determined by UV spectrophotometry and diluted to 30 ng/. mu.l.
SSR-PCR adopted a 10. mu.l reaction system: 25mmol/l Mg2+0.8. mu.l, 0.2. mu.l of 10. mu. mol/l primer, 0.3. mu.l of 10mmol/l dNTP, 0.05. mu.l of 5U/mu.l Taq enzyme, 2.00. mu.l of 5-10 ng/mu.l DNA template, 1.0. mu.l of 10 XPCR buffer, ddH2O5.45 μ l; the PCR reaction conditions are pre-denaturation at 94 ℃ for 3min, denaturation at 94 ℃ for 1min, annealing at 54-58 ℃ for 1min, and extension at 72 ℃ for l min for 35 cycles; finally, extension is carried out at 72 ℃ for 10min, and the reaction is terminated at 4 ℃ for 10 min.
And (3) detecting by polyacrylamide gel electrophoresis: PCR products were detected by 6% denaturing PAGE, and the results were visualized by silver nitrate staining after electrophoresis. Reading the amplified spectral bands according to the result of PAGE gel electrophoresis, and regarding each SSR amplification site as 1 gene site, wherein each site is marked as AA, BB, CC, AB, BC, AC and the like to form original data. The obtained SSR data are used for calculating the polymorphic site percentage (PPL), the Shannon information diversity index (I) and the Nei's gene diversity index (HE) by using POPGENE software.
As shown in Table 5, the number of amplified loci of each primer pair was varied from 76 to 187, with an allelic factor of 2 to 4 for each locus, and 49 alleles were detected in total, with an average allelic factor of 3.5 for each locus. The total number of bands amplified by 14 pairs of SSR primers in a parent population is 57, 4.07 bands are averaged for each pair of primers, 50 polymorphic bands are obtained, the percentage of polymorphic sites (PPL) is 87.72%, the effective allele index (Nei' S) is 2.6656, and the Shannon information diversity index (I) is 1.0666.
TABLE 5 SSR primer amplification site number and polymorphism information content
Genetic distance between parents (Dg) was calculated by SAS/STAT software using the Nei genetic distance equation, and the results are shown in Table 6: the genetic distance variation among 6 parents (R37, Lu wax No. 3, R36-1, Lu wax No. 4, J5 and J2) in the Fraxinus velutina species is 0.0565-0.9444, and the average genetic distance is 0.6472; the genetic distance between the Fraxinus velutina parents (R37, Lu wax No. 3 and R36-1) and the Fraxinus rhynchophylla parents (Lu wax 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 villosus and the parent of the fraxinus rhynchophylla is far greater than the genetic distance between the parents of the fraxinus villosus and the parent of the fraxinus rhynchophylla. The genetic distance between the parents of 10 hybrid combinations is greatly different, the genetic distance between R36-1 and Lu wax No. 5 is 1.7996 at most, and the genetic distance between R36-1 and Lu wax No. 4 is 0.0565 at least. Wherein the parent genetic distance of the R37 XU wax No. 5 combination and the U wax No. 3 XU wax No. 5 combination with remarkable heterosis is 1.1985-1.4277.
TABLE 6 genetic distance between parents
Example 3
And (4) carrying out correlation analysis on heterosis and parent genetic distance. Constructing a scatter diagram of the filial generation heterosis changing along with the parent combining ability and the genetic distance in Microsoft Office Excel, detecting the fitting degree of the filial generation heterosis and the parent combining ability and the genetic distance unary linear regression model through software, and establishing a corresponding unary linear regression equation. The genetic distance between parents was analyzed by using sppss21.0 statistical analysis software in association with the heterosis and the combining ability of the offspring, respectively, and the results are shown in table 7.
The SSR marker genetic distance has a significant positive correlation with the sum of the hybrid progeny tree high growth amount, the tree high hybrid vigor, the tree high male parent general combining ability and the tree high general combining ability, the tree high special combining ability, the correlation coefficient is between 0.642 and 0.703, and the correlation with the tree high female parent general combining ability is not significant. The correlation analysis of the SSR marker genetic distance and the genetic parameters of the breast diameter shows that the correlation is not obvious, the correlation coefficient is compared to find that the SSR marker genetic distance has the highest correlation coefficient with the special combining ability, and the male parent has the second highest combining ability. Comparing the SSR marker genetic distance with the correlation coefficients of the tree height and the breast diameter, finding that the correlation coefficients of the SSR marker genetic distance and the tree height and the breast diameter are the highest with the correlation coefficients of the common combining ability and the special combining ability of the male parent, wherein the correlation coefficients are between 0.544 and 0.703; however, the correlation coefficient of the general combining force with the female parent is lower between 0.241 and 0.244. Further by regression analysis it was found that: genetic distance between parents, tree height heterosis, breast diameter heterosis and parent GCAP1+P2And SCA showed a linear correlation (Table 7). The significance test result shows that the genetic distance of the parent, the tree height heterosis and the GCA of the parentP1+P2And SCA, while none of the other linear equation tests reached a significant or very significant level.
TABLE 7 correlation of white wax genetic distance with heterosis and combining ability
The correlation between the SSR genetic distance between parents and the growth traits of filial generations can be reflected more intuitively in a scatter diagram of the genetic distance between the parents and the growth traits of the filial generations (figure 1). As the genetic distance between parents increases, the tree is highly miscellaneousSpecies dominance, parental GCAP1+P2And SCA, and when the genetic distance is more than 1, the genetic distance tends to be positively related. The scatter diagram shows that the correlation coefficient of the genetic distance of the parents and the special combining ability of the tree heights is 0.4936 at most, the correlation coefficient of the genetic distance of the parents and the general combining ability of the tree heights and the male parent GCA is 0.4899, and the scatter diagram is matched with the regression straight line through comparison analysis. Further by regression analysis it was found that: genetic distance between parents, tree height heterosis, breast diameter heterosis and parent GCAP1+P2And SCA show linear correlation (Table 7). The significance test result shows that the genetic distance of the parent and the tree height heterosis, the parent GCAP1+P2And SCA, while none of the other linear equation tests reached a significant or very significant level.
The examples can obtain that the female parent 'R37', 'Lu wax No. 3' and the male parent 'Lu wax No. 5', have generally higher combining ability, wherein the R37 XLu wax No. 5 and the Lu wax No. 3 XLu wax No. 5 combined family line has the maximum special combining ability effect values on the height and the diameter of the tree, and the combined hybrid superiority is also strongest. The higher the general combining ability of the parent, the higher the effect value of the special combining ability, thereby showing that the general combining ability of the parent is one of the important factors influencing the heterosis of the combination. Therefore, parents with higher combining ability are selected for matching, and then a combination with high special combining ability is selected, so that the heterosis of the combination can be effectively improved, and the hybrid combination with strong heterosis is screened out.
The combining ability has strong prediction ability on the growth character expression of filial generation and the heterosis, and can be used as an important basis for matching parents and predicting the heterosis of the filial generation. The linear correlation result of genetic distance, heterosis and combining ability shows that GD (genetic distance) and filial generation growth character phenotype value, heterosis and parental GCAP1+P2(sum of general affinities of parents), GCAP2The male parent general combining ability and SCA (special combining ability) are in obvious positive correlation, GD and GCAP1+P2The correlation between the phenotype value of the offspring and the heterosis is the strongest, GCAP2Second, GCAP1Weakest, GCA is used in selective breedingP1+P2The sub-representative value and heterosis are predicted to be more than GCAP1And GCAP2Has better accuracy. Therefore, GCA can be preferentially adopted for parent matching of constructed measure forest in breedingP1+P2Predicting the phenotype value of the growth traits of the filial generation, and predicting the heterosis of the growth traits of the filial generation by adopting the SCA; and for the parent matching of the non-constructed survey forest, the phenotype value of the growth traits of the filial generation and the heterosis can be more accurately predicted based on certain level genetic distance analysis.
The results of the embodiment of the invention show that the tree height and the breast diameter heterosis are increased along with the increase of the genetic distance between parents, and when the genetic distance is more than 1, the positive correlation tends to be realized, and the heterosis with the tree height is obviously increased. I.e., the probability of obtaining strong heterosis by the hybridization combination with the parents having far genetic distance is higher. The practical possibility of adopting SSR molecular markers to carry out white wax genetic diversity analysis, heterosis prediction and the like is shown. However, the correlation coefficient between the genetic distance and the heterosis obtained in the research is 0.695, and the comparison research of the 10 combined heterosis and the genetic distance shows that the heterosis effect is stronger when the genetic distance is larger, but 2 hybrid combinations with the genetic distance ranging from 1.1985 to 1.4277 show strong heterosis. Therefore, parents with genetic distance within a certain range are matched, and the heterosis is more favorably expressed. Therefore, the parents with the genetic distance between the female parent and the male parent larger than 1 are screened, and the obvious heterosis can be obtained. Particularly, the invention carries out genetic distance analysis based on molecular level, adopts SSR markers to quickly screen out the hybridization combination with the genetic distance between parents of 1.1-1.5, can obtain the matching of the parents without constructing a progeny survey forest, has remarkable progeny heterosis, can greatly shorten the forest breeding period, and shortens the forest breeding period by 4-5 years.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Sequence listing
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SHANDONG HUABO GENETIC ENGINEERING Co.,Ltd.
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Claims (9)
1. An SSR molecular marker primer group for predicting white wax heterosis, which is characterized by comprising 14 pairs of primers with the nucleotide sequences shown as SEQ ID NO.1-SEQ ID NO. 28.
2. The SSR molecular marker primer group of claim 1 is applied to white wax fine variety breeding or parent selection.
3. The use of the SSR molecular marker primer set of claim 1 for white wax heterosis prediction.
4. A method of predicting white wax heterosis, comprising the steps of: carrying out PCR amplification on parent materials by using the primer group of claim 1, calculating the genetic distance between the parents by using a Nei genetic distance formula, and screening out the hybridization combination with the genetic distance between the parents being more than 1.
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 the white wax comprises Fraxinus velutina and Fraxinus americana.
7. The method of claim 4, wherein said heterosis comprises tree height and breast diameter.
8. The method of claim 4, wherein the PCR reaction program is pre-denaturation at 94 ℃ for 3min, denaturation at 94 ℃ for 1min, annealing at 54-58 ℃ for 1min, and extension at 72 ℃ for l min for 35 cycles; finally, extension is carried out at 72 ℃ for 10min, and the reaction is terminated at 4 ℃ for 10 min.
9. The method of claim 4, wherein the reaction system of the PCR comprises the following components in parts by weight: 25mmol/LMg2+0.8 part, 0.2 part of primer of 10 mu mol/L, 0.3 part of LdNTP of 10mmol/L, 0.05 part of 5U/mu LTaq enzyme, 2.00 parts of DNA template of 5-10 ng/mu L, 1.0 part of 10 XPCR buffer solution, ddH2And 5.45 parts of O.
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Citations (4)
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| 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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| 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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