CN102146436B - Method for assisting in screening of salt-resistance soybeans and special primers for the same - Google Patents
Method for assisting in screening of salt-resistance soybeans and special primers for the same Download PDFInfo
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- CN102146436B CN102146436B CN 201010110877 CN201010110877A CN102146436B CN 102146436 B CN102146436 B CN 102146436B CN 201010110877 CN201010110877 CN 201010110877 CN 201010110877 A CN201010110877 A CN 201010110877A CN 102146436 B CN102146436 B CN 102146436B
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Abstract
The invention discloses a method for assisting in screening of salt-resistance soybeans and special primers for the same. The special primers provided by the invention are a specific primer pair consisting of nucleotide represented by a sequence 3 and a sequence 4 in a sequence table. The method provided by the invention comprises the following steps: performing PCR amplification by using the genome DNA soybeans to be tested as a template and by using the specific primer pair, and detecting the products of the amplification, wherein soybeans to be tested, which have 383bp fragments, among the amplification products are salt-resistance soybeans, and soybeans to be tested, which have no 383bp fragment, among the amplification products are salt sensitive soybeans. When the specific primer pair disclosed by the invention is used to assist in screening salt-resistance soybeans or salt-sensitive soybeans, the operation is simple and convenient, and the correction rate is high. The method and the primers can be used in identification of the salt-resistance property of the soybeans and can be used for early screening. When the method and the specific primer pair are used in breeding, the breeding time is reduced, the breeding cost is saved, the breeding success rate is increased and a great practical value is achieved.
Description
Technical field
The present invention relates to a kind of method and primer special thereof of assisting in screening of salt-resistance soybeans.
Background technology
Soybean belongs to the moderate salt-tolerant plant, since the eighties in last century, Shao Guihua etc. carry out Salt-Tolerance Identification to nearly more than the 6000 parts of soybean resources of China, screen bud phase height Salinity tolerance germplasm (1,2 grade) 49 parts, 34 parts of Salt Tolerance at Seedling Stage germplasms, also identify in addition Salinity tolerance germplasm in some (Shao Guihua ，Chang Ru town, an old dance in the time of infertility, soybean salt-tolerance Journal of Sex Research progress. Soybean Science .1993,12 (3): 244-248).Soybean different growing salt tolerance difference, be subject to salt damage larger on the impact of output, therefore seedling stage, the Salt-Tolerance Identification in seedling stage particularly important (Shao Guihua etc., Song Jingzhi, Liu Huiling. Preliminary Studies On The Evaluation of Salt Tolerance In Soybean Varieties. Scientia Agricultura Sinica, 1986, (6): 30-35).Guo Bei etc. utilize the rich No. 7 * UnionF of literary composition
2the proof soybean salt-tolerance gene is that pair of dominant genes is controlled, utilize the RAPD technology, identify the RAPD mark (Guo Bei of Salt Tolerance at Seedling Stage gene in the salt tolerant kind in the time of infertility, Qiu Lijuan, Shao Guihua, the Chang Ru town, Liu Lihong, Xu Zhanyou, Li Xianghua, Sun Jianying. the PCR mark of soybean salt-tolerance gene. Scientia Agricultura Sinica, 2000, 33:10-16), and carry out the screening operation (Guo Bei of salt tolerant resource with this mark, Qiu Lijuan, Shao Guihua, Xu Zhanyou. the molecule marker assistant identification of soybean salt-tolerance gene germplasm and utilization research thereof. Soybean Science, 2002, 21:56-61).Utilize cultivation and wild soybean colony to carry out the discovery of soybean salt-tolerance Journal of Sex Research, a main effect QTL with the soybean salt-tolerance height correlation is arranged between soybean N linkage group same zone, can explain 68.7% of heritable variation at wild soybean, 69% of soluble heritable variation (Lee GJ in the cultivated soybean, Boerma HR, Villagarcia MR, Zhou X, CarterTE, Li Z, Gibbs MO.A major QTL conditioning salt tolerance in S-100 soybeanand descendent cultivars.Theor Appl Genet, 2004, 109:1610-1619, Hamwieh A, Xu DH.Conserved salt tolerance quantitative trait locus (QTL) in wild andcultivated soybeans.Breeding Sci, 2008,58:355-359), illustrate in the cultivated soybean and wild soybean and may have common resistant gene of salt.
Summary of the invention
The method and the primer special thereof that the purpose of this invention is to provide a kind of assisting in screening of salt-resistance soybeans.
The Auele Specific Primer pair that primer special provided by the invention is comprised of Nucleotide shown in the sequence 4 of the sequence 3 of sequence table and sequence table.
The method of assisting in screening of salt-resistance soybeans provided by the invention comprises the steps (a) or step (b):
(a) take the genomic dna of soybean to be measured is template, with described Auele Specific Primer, (primer special) is carried out to pcr amplification, detects amplified production, has the salt tolerant soybean that the soybean to be measured of 383bp fragment is the candidate in amplified production;
(b) detect the DNA fragmentation shown in the sequence 1 that whether there is sequence table in the genomic dna of soybean to be measured, the salt tolerant soybean that the soybean to be measured that has the DNA fragmentation shown in the sequence 1 of sequence table in genomic dna is the candidate.
Specifically, in described step (a), there is the 383bp fragment in amplified production and do not there is the salt tolerant soybean that the soybean to be measured of 263bp fragment is the candidate.Specifically in described step (b), there is the DNA fragmentation shown in the sequence 1 of sequence table in genomic dna and do not there is the salt tolerant soybean that the soybean to be measured of the DNA fragmentation shown in the sequence 2 of sequence table is the candidate.
The method of the responsive soybean of assisting sifting salt provided by the invention comprises the steps (c) or step (d):
(c) take the genomic dna of soybean to be measured is template, with described Auele Specific Primer, (primer special) carried out to pcr amplification, detect amplified production, there is the 263bp fragment in amplified production and do not there is the responsive soybean of salt that the soybean to be measured of 383bp fragment is the candidate;
(d) detect in the genomic dna of soybean to be measured whether there is the sequence 1 of sequence table and the DNA fragmentation shown in sequence 2, there is the DNA fragmentation shown in the sequence 2 of sequence table in genomic dna and do not there is the responsive soybean of salt that the soybean to be measured of the DNA fragmentation shown in the sequence 1 of sequence table is the candidate.
It is the soybean below 2 that described salt tolerant soybean specifically can be the salt tolerant rank.It is the soybean more than 3 that the responsive soybean of described salt specifically can be the salt tolerant rank.
The method of assistant identification soybean salt-tolerance proterties provided by the invention comprises the steps (e) or step (f):
(e) take the genomic dna of soybean to be measured is template, with described Auele Specific Primer, (primer special) is carried out to pcr amplification, detects amplified production; The salt tolerant rank that has the soybean to be measured of 383bp fragment in amplified production is below 2; The salt tolerant rank that has the 263bp fragment in amplified production and do not have a soybean to be measured of 383bp fragment is more than 3;
(f) detect in the genomic dna of soybean to be measured whether there is the sequence 1 of sequence table and the DNA fragmentation shown in sequence 2; The salt tolerant rank that has the soybean to be measured of the DNA fragmentation shown in the sequence 1 of sequence table in genomic dna is below 2; The salt tolerant rank that has the DNA fragmentation shown in the sequence 2 of sequence table in genomic dna and do not have a soybean to be measured of the DNA fragmentation shown in the sequence 1 of sequence table is more than 3.
Specifically, in described step (e), the salt tolerant rank that has the 383bp fragment in amplified production and do not have a soybean to be measured of 263bp fragment is below 2.Specifically, in described step (f), the salt tolerant rank that has the DNA fragmentation shown in the sequence 1 of sequence table in genomic dna and do not have a soybean to be measured of the DNA fragmentation shown in the sequence 2 of sequence table is below 2.
Described salt tolerant is mainly reflected on Salt Tolerance at Seedling Stage.Described salt sensitivity is mainly reflected on salt sensitivity in seedling stage.Other division of salt tolerant level can be with reference to the authentication method (Shao Guihua etc. of Shao Guihua etc., Song Jingzhi, Liu Huiling. Preliminary Studies On The Evaluation of Salt Tolerance In Soybean Varieties. Scientia Agricultura Sinica, 1986, (6): 30-35): according to the individual plant leaf area, divide division as follows the salt tolerant rank of soybean (salt tolerant classification): 0 grade: plant strain growth is normal, blade edge, without the salt damage symptom; 1 grade: plant strain growth is normal, the leaf area of being injured<10% or 4 greenery are arranged; 2 grades: plant strain growth normal, the leaf area of being injured 25% or 3 greenery are arranged; 3 grades: plant strain growth is suppressed, the leaf area of being injured 50% or 2 greenery are arranged; 4 grades: plant strain growth is seriously suppressed, the leaf area of being injured 75% or 1 greenery is arranged; 5 grades: plant is dead or the lobus cardiacus survival is only arranged, and is injured leaf area more than 75%.
Specifically, the reaction system of described pcr amplification is: 1 * PCR Buffer, 1.5mmol/L MgCl
2, 150 μ mol/L dNTPs, the 150pmol/L upstream primer, the 150pmol/L downstream primer, 1U Taq enzyme, the 40ng genomic dna, add water and supply 20 μ l.Specifically, the response procedures of described pcr amplification is: 94 ℃ of denaturation 5min; 94 ℃ of sex change 30sec, 55 ℃ of annealing 30sec, 72 ℃ are extended 1min, move 35 circulations; Last 72 ℃ are extended 5min.
Described Auele Specific Primer is to also belonging to protection scope of the present invention being prepared as follows three kinds of application in test kit: the test kit of (1) assisting in screening of salt-resistance soybeans; (2) test kit of the responsive soybean of assisting sifting salt; (3) test kit of assistant identification soybean salt-tolerance proterties.
The present invention protects following three kinds of test kits simultaneously: the test kit of (1) assisting in screening of salt-resistance soybeans comprises described Auele Specific Primer pair; (2) test kit of the responsive soybean of assisting sifting salt, comprise described Auele Specific Primer pair; (3) test kit of assistant identification soybean salt-tolerance proterties, comprise described Auele Specific Primer pair.
The present invention also protects the DNA molecular relevant to the soybean salt-tolerance proterties, is the DNA fragmentation shown in the sequence 2 of the DNA fragmentation shown in the sequence 1 of sequence table or sequence table.Described DNA molecular is a kind of SCAR mark (sequencecharacterized amplified region).
Apply Auele Specific Primer of the present invention to the responsive soybean of assisting in screening of salt-resistance soybeans or salt, easy and simple to handle, accuracy is high.The present invention can be applied to the evaluation of soybean seedling salt-tolerance character, can be in office when the phase is extracted Soybean genomic DNA is screened.Method of the present invention and Auele Specific Primer, to for the salt tolerant soybean breeder, can be shortened to breeding time, save the breeding cost, increase and be bred as success ratio, there is great actual value.
The accompanying drawing explanation
Fig. 1 is rich No. 7 and Union distinguished sequence difference of literary composition; T: rich No. 7 of literary composition; S:Union.
The electrophorogram of the amplified production that Fig. 2 is literary composition rich No. 7 and Union; M:marker; 1: rich No. 7 of literary composition; 2:Union.
Fig. 3 is the electrophorogram of the specific fragment of the literary composition reclaimed rich No. 7 and Union; 1: rich No. 7 of literary composition; 2:Union.
Fig. 4 is rich No. 7 of literary composition, Union and part F
2electrophorogram for the individual plant pcr amplification product; P1: rich No. 7 of literary composition; P2:Union; 1-15: part F
2for individual plant.
Fig. 5 is Auele Specific Primer to 20 parts of salt tolerant soybean and the 17 parts of responsive soybean amplified production of salt electrophorograms; The swimming lane numbering is corresponding with the sequence number in table 1.
Embodiment
Following embodiment is convenient to understand better the present invention, but does not limit the present invention.Experimental technique in following embodiment, if no special instructions, be ordinary method.Test materials used in following embodiment, if no special instructions, be and purchase available from routine biochemistry reagent shop.
Rich No. 7 of soybean literary composition: derive from country of Chinese Academy of Agricultural Sciences germplasm resource bank, numbering ZDD02611 in storehouse; 1971 by the Jining City in Shandong Province area institute of agricultural sciences by the sexual hybridization seed selection.
Soybean Union: derive from country of Chinese Academy of Agricultural Sciences germplasm resource bank, numbering WDD00617 in storehouse; It is the soybean varieties that Inst., of Breeds of Crops, Chinese Academy of Agriculture introduces from the U.S. in 1984.
Reclaim respectively specific fragment from literary composition rich No. 7 (salt tolerant soybean) and soybean Union (the responsive soybean of salt), and checked order.Obtain the sequence of 767bp in Salt-tolerant Materials, obtained the sequence of 655bp in the salt sensitive material.The pcr amplification of two kind soybean the results are shown in Figure 2, and the electrophorogram of specific fragment is shown in Fig. 3.Order-checking and the comparison result of two specific fragments are shown in Fig. 1.
Sequence alignment is found: Salt-tolerant Materials, than the insertion sequence that a 130bp is arranged in salt sensitive material, also has in addition 3 and deletes fragment (being respectively 10bp, 6bp and 2bp)), also have 2 SNP (1 replacement, 1 transversion).Utilize the conservative region exploitation Auele Specific Primer pair of two sections sequences, the insertion of detection 130bp and 10bp/deletion polymorphism, thus obtain codominant marker's (SCAR mark).
Auele Specific Primer pair:
Upstream primer (21 Nucleotide): 5 '-ACGTAAGTGGTTGAAGGCGTT-3 ' (sequence 3 of sequence table);
Downstream primer (18 Nucleotide): 5 '-GGGCAAGGGATATGAAAA-3 ' (sequence 4 of sequence table).
The association analysis of embodiment 2, specific band and soybean salt-tolerance
One, the rich No. 7 * Union F of literary composition
2the acquisition of colony and Salt-Tolerance Identification
By the rich F obtained with Union hybridization for No. 7 of literary composition
1the plant selfing, obtain the F be comprised of 195 individual plants
2colony.This colony was sowed on May 20th, 1997, and in Shandong Province, Changyi county stud farm carries out the salt tolerance field test.Experimental field belonging to the strong sandy saline-alkali soil of moderate salinization, is the inwelling district, and soil salt content is 0.4%～0.7%.Plant strain growth is to V3 (2～3 compound leaves launch) during period, and it is 14.8dsm that extracting saline groundwater (its composition forms identical with seawater) is deployed into concentration with ground fresh water
-1(saltiness 0.95%; The quality percentage composition) salt water, processed soybean plant strain.The rear 3d of processing has or not according to salting stain on plant leaf, salt tolerance and the brine sensitivity of investigation record individual plant, and growing the later stage, further determine its plant salt tolerant phenotype.F
2it is 148 strains that Salt-tolerant Materials is arranged in colony, and the salt sensitive material is 47 strains.
Two, the association analysis of specific band and soybean salt-tolerance
Sample: rich No. 7 of literary composition, Union and 195 F
2individual plant.
Extract respectively the genomic dna of each sample, the genomic dna of take utilizes the Auele Specific Primer of embodiment 1 exploitation to carrying out pcr amplification as template.
The PCR reaction system: 1 * PCR Buffer, 1.5mmol/L MgCl
2, 150 μ mol/L dNTPs, the 150pmol/L upstream primer, the 150pmol/L downstream primer, 1U Taq enzyme, the 40ng genomic dna, add redistilled water and supply 20 μ l.
PCR response procedures: 94 ℃ of denaturation 5min; 94 ℃ of sex change 30sec, 55 ℃ of annealing 30sec, 72 ℃ are extended 1min, move 35 circulations; Last 72 ℃ are extended 5min.
Reaction adds 6 μ l Loading Buffer after finishing in amplified production, electrophoresis on 2% agarose, each point sample hole loading 10 μ l, 100V constant voltage electrophoresis 40min, scanning under gel imaging system, statistics.
The literary composition banding pattern of rich No. 7: there is the band of 383bp and do not there is the band of 263bp.The banding pattern of Union is: have the band of 263bp, and do not have the band of 383bp.In 195 F2 individual plants, 34 have and the rich No. 7 same banding patterns of literary composition, and 47 have the banding pattern same with Union, 114 remaining bands (heterozygosis banding pattern) that simultaneously have 263bp and 383bp, this has the individual plant of 2 bands simultaneously, is the heterozygote generation of rich No. 7 and Union of literary composition.In 148 salt tolerant individual plants, 34 individual plants show and the rich No. 7 identical banding patterns of literary composition, and 113 individual plants show the heterozygosis banding patterns, 1 individual plant demonstration Union banding pattern.In 47 responsive individual plants of salt, 46 individual plants show banding pattern identical with Union, and 1 individual plant shows the heterozygosis banding pattern.The pcr amplification of part sample the results are shown in Figure 4.
Results suggest: show and the individual plant of civilian rich No. 7 identical banding patterns, one is decided to be the salt tolerant individual plant; Showing the individual plant of heterozygosis banding pattern, is probably the salt tolerant individual plant; Showing the individual plant with the identical banding pattern of Union, is probably salt sensitivity individual plant.
Find two exchange individual plants in Salt-tolerant Materials, the genetic distance of this mark and resistant gene of salt is 1cM.
1/2 hoagland nutritive medium preparation method is as follows: solvent is distilled water, and solute and concentration thereof are as follows: 2mM Ca (NO
3)
24H
2o, 2.5mM KNO
3, 0.5mM NH
4nO
3, 1mM KH
2pO
4, 1mM MgSO
4, 0.1mM FeSO
47H
2o, 0.1mM EDTA-Na
2, 0.025mM H
3bO
3, 0.002mM MnSO
4h
2o, 0.002mM ZnSO
47H
2o, 0.0005mMNa
2moO
42H
2o, 0.0005mM CuSO
45H
2o and 0.001mM CoCl
2.
One, the Salt-Tolerance Identification of each kind soybean
The soybean of 37 kinds, all derive from country of Chinese Academy of Agricultural Sciences germplasm resource bank, and in Germplasm Bank, numbering and salt tolerance are in Table 1.
Salt-Tolerance Identification method (germ plasm resource water culture): identify and carry out in the solarium of Institute of Crop Science, Chinese Academy of Agricultural Science; Seed is sprouted in the germination box of vermiculite is housed, and when cotyledon launches, the seedling of neat and consistent is moved into to the 1/2hoagland nutritive medium, and each kind is established 3 repetitions, and each repeats 5 strains.Every 3 days of nutritive medium is changed once, when being cultured to the seedling true leaf and launching fully, is replaced by the identical nutritive medium that contains 100mM NaCl and carries out Stress treatment.Investigate each individual plant salt tolerant rank during 10d, take the mean as the salt tolerance of each kind.Other divides the authentication method (Shao Guihua etc. with reference to Shao Guihua etc. the salt tolerant level, Song Jingzhi, Liu Huiling. Preliminary Studies On The Evaluation of Salt Tolerance In Soybean Varieties. Scientia Agricultura Sinica, 1986, (6): 30-35): according to the individual plant leaf area, divide division as follows the salt tolerant rank of soybean (salt tolerant classification): 0 grade: plant strain growth is normal, blade edge, without the salt damage symptom; 1 grade: plant strain growth is normal, the leaf area of being injured<10% or 4 greenery are arranged; 2 grades: plant strain growth normal, the leaf area of being injured 25% or 3 greenery are arranged; 3 grades: plant strain growth is suppressed, the leaf area of being injured 50% or 2 greenery are arranged; 4 grades: plant strain growth is seriously suppressed, the leaf area of being injured 75% or 1 greenery is arranged; 5 grades: plant is dead or the lobus cardiacus survival is only arranged, and is injured leaf area more than 75%.The material of salt tolerant rank >=3 is as the salt sensitive material, salt tolerant rank≤2 as Salt-tolerant Materials.
Table 1 is for molecule marker aided verification soybean resource title and salt tolerance
Result shows, 37 kinds comprise 20 parts of salt tolerant kinds and 17 parts of salt sensitive varietieies.
Two, the application specific primer pair carries out somatotype
With this SCAR mark, 20 salt tolerant soybean resources and 17 parts of responsive soybean resources of salt are detected.Extract respectively the genomic dna of each sample, the genomic dna of take utilizes the Auele Specific Primer of embodiment 1 exploitation to carrying out pcr amplification as template.The PCR reaction system: 1 * PCR Buffer, 1.5mmol/L MgCl
2, 150 μ mol/L dNTPs, the 150pmol/L upstream primer, the 150pmol/L downstream primer, 1U Taq enzyme, the 40ng genomic dna, add redistilled water and supply 20 μ l.PCR response procedures: 94 ℃ of denaturation 5min; 94 ℃ of sex change 30sec, 55 ℃ of annealing 30sec, 72 ℃ are extended 1min, move 35 circulations; Last 72 ℃ are extended 5min.Reaction adds 6 μ lLoading Buffer after finishing in amplified production, electrophoresis on 2% agarose, each point sample hole loading 10 μ l, 100V constant voltage electrophoresis 40min, scanning under gel imaging system, statistics.The electrophorogram of each sample is shown in Fig. 5.Repeat three experiments, result is consistent.
20 parts of salt tolerant kind banding patterns are as follows: have the band of 383bp and do not have the band of 263bp.In 17 parts of salt sensitive varietieies: 15 parts of banding patterns are as follows: have the band of 263bp, and do not have the band of 383bp; Other two parts of demonstrations banding pattern identical with the salt tolerant kind.Result shows only to have amplified in two parts of salt sensitive varietieies the peculiar banding pattern of salt tolerant, and other materials genotype and phenotype are in full accord, and identical rate is 94.9%.Therefore, this mark can be widely used in the Salt-Tolerance Identification of Soybean Germplasm.
Results suggest: have the band of 383bp and do not have the soybean of the band of 263bp, one is decided to be the salt tolerant soybean; Having the band of 263bp and do not have the soybean of the band of 383bp, is probably the responsive soybean of salt.
Three, sequence verification
The PCR product is checked order respectively, and result shows: the Nucleotide shown in the sequence 1 that the PCR product of 20 parts of salt tolerant soybean is sequence table; In the responsive soybean of 17 portions of salt, the Nucleotide shown in the sequence 2 that 15 parts of PCR products with soybean of identical banding pattern are sequence table, the Nucleotide shown in the sequence 1 that the PCR product of the soybean of the banding pattern that 2 parts of demonstrations are identical with the salt tolerant kind is sequence table.
Sequence table
<110 > Institute of Crop Science, Chinese Academy of Agricultural Science
<120 > a kind of method of assisting in screening of salt-resistance soybeans and primer special thereof
<130>CGGNARY102095
<160>4
<210>1
<211>383
<212>DNA
<213 > Glycine soybean (Glycine max (L.))
<400>1
acgtaagtgg ttgaaggcgt taaccggtga tgattaaata ccccttcccc ttctaaataa 60
aggccattta taacctcaaa tttatacttt tctttccgta tacttcagac gtaattaata 120
attattaact ttcttctttt tattattatt tttcttgcct tattataact gttgtgtaaa 180
aaaaaaatta cctcaaaata attatcattt taaattttta tttatatttc ttataatatt 240
aataataaat aaaattataa atatattaat aataatataa tgttaatttt gtaatttttt 300
tatctattta ttatattttt tatttatata aaataattga agaagataac agaaaaataa 360
tatttttttc atatcccttg ccc 383
<210>2
<211>263
<212>DNA
<213 > Glycine soybean (Glycine max (L.))
<400>2
acgtaagtgg ttgaaggcgt taacagtgtt tcaccggtga tgattaaata ccccttcccc 60
ttctaaataa aggccattta taacctcatt ttaaattttt atttatattt cttataatat 120
taataataaa taaaattata aatatattaa taataatata atgttaattt tgtaattttt 180
ttatctattt attatttttt tatttatata aaataattga agaagataac agaaaaataa 240
tatttttttc atatcccttg ccc 263
<210>3
<211>21
<212>DNA
<213 > artificial sequence
<220>
<223>
<400>3
acgtaagtgg ttgaaggcgt t 21
<210>4
<211>18
<212>DNA
<213 > artificial sequence
<220>
<223>
<400>4
Claims (5)
1. the Auele Specific Primer pair formed by Nucleotide shown in the sequence 4 of the sequence 3 of sequence table and sequence table.
2. the method for an assistant identification soybean salt-tolerance proterties comprises the steps (e) or step (f):
(e) take the genomic dna of soybean to be measured is template, with Auele Specific Primer claimed in claim 1, to carrying out pcr amplification, detects amplified production; There is the 383bp fragment in amplified production and do not have the 263bp fragment the salt tolerant rank of soybean to be measured be below 2; The salt tolerant rank that has the 263bp fragment in amplified production and do not have a soybean to be measured of 383bp fragment is more than 3;
(f) detect in the genomic dna of soybean to be measured whether there is the sequence 1 of sequence table and the DNA fragmentation shown in sequence 2; The salt tolerant rank that has the DNA fragmentation shown in the sequence 1 of sequence table in genomic dna and do not have a soybean to be measured of the DNA fragmentation shown in the sequence 2 of sequence table is below 2; The salt tolerant rank that has the DNA fragmentation shown in the sequence 2 of sequence table in genomic dna and do not have a soybean to be measured of the DNA fragmentation shown in the sequence 1 of sequence table is more than 3.
3. the described Auele Specific Primer of claim 1 is to being prepared as follows three kinds of application in test kit:
(1) test kit of assisting in screening of salt-resistance soybeans;
(2) test kit of the responsive soybean of assisting sifting salt;
(3) test kit of assistant identification soybean salt-tolerance proterties.
4. the test kit of the responsive soybean of the test kit of assisting in screening of salt-resistance soybeans and/or assisting sifting salt and/or the test kit of assistant identification soybean salt-tolerance proterties, comprise the described Auele Specific Primer pair of claim 1.
5. the DNA molecular relevant to the soybean salt-tolerance proterties is the DNA fragmentation shown in the DNA fragmentation shown in sequence 1 or sequence 2 in sequence table.
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| CN1258746A (en) * | 1998-12-30 | 2000-07-05 | 中国农业科学院作物品种资源研究所 | Molecular mark of soybean salt-tolerance gene and its acquiring method and application |
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| CN1258746A (en) * | 1998-12-30 | 2000-07-05 | 中国农业科学院作物品种资源研究所 | Molecular mark of soybean salt-tolerance gene and its acquiring method and application |
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| Title |
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| Wang D et al.Emergence and seedling growth of soybean cultivars and maturity groups under salinity groups under salinity.《 Plant Soil》.1999,全文. * |
| 张海燕.大豆耐盐基因定位及耐盐相关基因分子标记的开发.《中国优秀硕士学位论文全文数据库》.2005,全文. * |
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