CN107043813A - No. 25 chromosomes of upland cotton SNP marker related to fibre strength - Google Patents

No. 25 chromosomes of upland cotton SNP marker related to fibre strength Download PDF

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CN107043813A
CN107043813A CN201710097667.1A CN201710097667A CN107043813A CN 107043813 A CN107043813 A CN 107043813A CN 201710097667 A CN201710097667 A CN 201710097667A CN 107043813 A CN107043813 A CN 107043813A
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商海红
巩万奎
范森淼
张震
袁有禄
石玉真
吴东
葛群
刘爱英
龚举武
李俊文
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Institute of Cotton Research of Chinese Academy of Agricultural Sciences
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Abstract

No. 25 chromosomes of upland cotton SNP marker related to fibre strength.The invention belongs to Molecular breeding in upland cotton technical field, a kind of SNP marker related to upland cotton fiber intensity and its detection and application are disclosed.Described SNP marker is that the method for resurveying sequence by genome is obtained using the stable RIL colonies of cotton as material.Using these SNP markers disclosed in this invention, molecular mark selection is carried out, breeding cycle is substantially shorter, the breeding efficiency of cotton fiber strength is improved.

Description

No. 25 chromosomes of upland cotton SNP marker related to fibre strength
Technical field
The invention belongs to Molecular breeding in upland cotton technical field, and in particular to a kind of SNP related to upland cotton fiber intensity Molecular labeling and its detection and application.
Background technology
Cotton occupies an important position as a kind of main fibre crops in World Economics.Planted extensively in the whole world In four cotton seeds of training, upland cotton accounts for more than the 90% of World cotton total output in occupation of most important status, its yield.With The growth of people's centering grade textiles demand, the requirement to fiber quality is increasingly improved, but traditional breeding technique is mainly Selected by phenotype, breeding efficiency is low, it is difficult to the need for meeting quality breeding.The development of molecular marking technique makes direct choosing Selecting the genotype of quantitative character becomes possibility.Breeder can be made direct by building the progress QTL positioning of cotton genetic map The genotype of the quantitative characters such as fiber quality is selected, by using F2Fiber quality QTL positioning has been carried out with the mapping population such as RIL Research also achieves great successes.More can be later mark especially with the development and application of third generation labelling technique SNP marker Note assistant breeding lays the first stone.SNP marks are current most potential molecular labelings, because quantity is more in genome, point Cloth is wide and need not be suitable for large-scale automation and quantity according to clip size by DNA points of bands during genetic analysis Huge detection and analysis, has been widely used in the field such as medical science and biology at present.But the research in cotton is also less.
SNP is that organism is most universal, the widest polymorphic difference of distribution.To the gene of 3 natural strains of arabidopsis Group sequencing and resurvey sequence, it has been found that the SNP sites more than 820,000, average about every 1.4 kb just has 1 SNP. The hereditary difference represented between individual of SNP high degrees, is remarkably contributing to carry out special quantitative inheritance character and individual point Son mark correlation analysis (Ossowski S., Schneeberger K., Clark R.M., Lanz C., Warthmann N., and Weigel D., Sequencing of natural strains ofArabidopsis thaliana with short reads, Genome Res., 2008,18 (12):2024-2033);Chu Ying etc. passes through fibre The SNP research and utilization round pcrs of dimension development related gene have separated the tetraploid cotton seed material and two of seven different fiber qualities The Fibre Development related gene fragment of individual diploid progenitors cotton seed, obtains five cotton fiber development related genes in nine cottons altogether The sequence information in material is planted, the SNPs sites upland cotton high-intensity fiber strain are counted, discovery there are four cSNPs There is nonsynonymous mutation in site, there may be phenotypic effect（Chu Ying, SNP researchs and the cotton sugarcane of Fibre Development related gene The clone of sugared synthase gene fragment and expression analysis, master's thesis, Nanjing:Agricultural University Of Nanjing, 2004）；Zheng Weijia etc. By carrying out SSR and SNP marker primer screening in new sea 21 and new land 36, two F of the primer pair filtered out are used2Group Body enters performing PCR amplification, builds genetic linkage mapses, 69 pairs of obtained SSR primers and 11 groups of SNP primer pairs F are screened in colony2 Colony amplifies the polymorphism mark come and carries out genetic linkage analysis, construct genetic linkage map including 90 marker sites and Average distance between 30 linkage groups, mark is 55.9cM, and total length is 3135.84cM, the 62.71% of covering cotton gene group（Zheng It is bright good, the structure of the exploitation of SNP marker and extra large land genetic linkage mapses, Master's thesis, Urumchi:Xinjiang Agricultural Univ, 2013）；He Linchi etc. is discussion affiliation nearer salt tolerant cotton variety CCRI 35 and salt density value cotton variety CCRI 12 salt tolerant othernesses, handle cotton seedling with various concentrations NaCl, investigate this 2 cotton varieties hydrogen peroxide under salt stress The change of the activities of antioxidant enzymes such as enzyme, peroxidase, superoxide dismutase, and to CCRI 35 and CCRI 12 SNP/In-Del carries out Preliminary Identification, it is found that affiliation is close and the significant CCRI 35 of salt tolerant level difference and CCRI 12 have point mutation and insertion/deletion, illustrate that these SNP/InDel may have important adjusting function to salt-tolerance character（What Lin Chi, Wang Kang, Wei little Yun, etc. the activities of antioxidant enzymes of salt tolerant otherness difference cotton variety and SNP/InDel analyses, river Su Nongye journals, 2014,30 (5):980-985）；Zhu constructs full-length genome SNP linkage maps by recombinant inbred lines Spectrum, finds 2618 polymorphism SNP markers, wherein with the presence of in 16 stable two environment of QTLs, 12 QTL are related to many property Shape, these QTLs are mainly distributed on 5,9,10,14,19, and No. 20 chromosomes（Li C,Zhu SJ et al.Genome- Wide SNP Linkage Mapping and QTL Analysis for Fiber Quality and Yield Traits in the Upland Cotton Recombinant Inbred Lines Population.Frontiers in Plant Science,2016,7:218）；Kuang Meng etc. using representational material carry out the genome-wide screening analysis in SNP sites with Comprehensive assessment, based on KASP technological development, 1 set is applied to the core SNP groups that Cotton in China purity of hybrid high flux is detected Close, from 63K cotton full-length genome chip screening obtain the SNP marks with single copy characteristic be respectively 5474 (in Cotton institute TM-1 genomes version) and 1850 (Nanjing agricultural university TM-1 genomes version), finally from every chromosome preferably 1 cenospecies heterozygosis rate height and the preferable core SNP sites of parting effect, add up to 26, will using KrakenTM softwares SNP sites change into KASP marks, carry out SNP parting detections using SNPline platforms, realize the height to a large amount of samples Flux gene parting, the variety that is particularly suitable for use in quick detection is that SNP labelling techniques are identified and fingerprint number in cotton variety Application in terms of being built according to storehouse lays the foundation（Kuang Meng, Ma Zhiying etc., the cotton crossbreed purity based on single copy SNP marks High flux detection technique, Cotton Science Cotton Science 2016,28 (3):227―233）.
Yuan Youlu etc. is built using a Gossypium anomalum high-intensity fiber Introgressed line 7235 and upland cotton Genetic standard line TM-1 for parent F2、F2：3Segregating population, master can be detected in multiple environment such as the different cotton regions and the U.S. of China by identifying one QTL is imitated, more than 30% phenotypic variation can be explained（Yuan Youlu etc., cotton high quality fiber character QTLs molecular marker screening and It is positioned, Acta Genetica Sinica, 2001,28 (12):1151-1161)；Shen Xinlian etc. utilizes 3 upland cotton high-intensity fiber germplasm line structures 3 F are built2Linkage map in kind, detects 38 QTLss relevant with fiber quality, wherein 15 using composite interval mapping method Stable QTLs can be simultaneously in F2And F2:3Detect, at least three QTLs can be expressed in Liang Ge colonies（Shen Xinlian, upland cotton is fine Tie up quality QTL screening, positioning and its application, doctor's thesis, Agricultural University Of Nanjing, 2004)；Beautiful true wait of stone is flowed with the Yellow River Transgenic Cotton Varieties sGK321 and sGK9708 that domain is planted extensively (in 41) are recurrent parent, respectively with harvesting high-quality high-yield product Kind too 121 and high microsteping quality gradually ooze the F of the hybridization of germplasm line 72351Hybridize for material and be returned, be configured with hybridization backcrossing combination Two sets, with 2 SSR markers with an oriented high-intensity fiber QTL close linkage, this 2 are marked at different heredity Background, after excessive generation hybridization, backcrossing and selfing, the effect that can stablize heredity and QTL is stable, and with technique knot Close other means and carry out high-quality, pest-resistant isogenic pyramiding breeding research, fast and effeciently improve existing upland cotton descriptive literature Kind, create high yield, high-quality, pest-resistant cotton new material or new lines (stone beautiful and the chain main effect QTL of cotton fiber strength such as true Applied to cotton molecular mark Molecular Plant Breedings, 2007,5 (4):521-527)；Wang Yiqing etc. utilizes upland cotton It is that male parent constructs F that Shandong cotton, which grinds 21 as maternal, NM03102,2And F2∶3Segregating population, to 195 F2Colony's individual plant analysis is obtained 242 marker sites, detect 20 fiber quality characteristics QTL altogether, wherein QTL and 1 micron value of 1 fibre strength QTL is stabilized in two generations, and this lays a good foundation for marker assisted selection（Wang Yiqing, Li Junwen, Yuan Youlu etc., land Cotton high-quality strain fiber quality characteristics QTL molecular labeling and positioning, Cotton Science, 2010,22 (6):533-538）；Jia Fei Deng utilization using SGK9708 as female parent, 0-153 is 196 upland cotton RIL (F6 that male parent is built:8) construct and include Average distance 4.45cM between 186 marks, overall length 827.84cM, mark, the genetic linkage map of covering cotton gene group 18.6% Spectrum, and QTL positioning and epistatic interaction analysis are carried out to the bell weight and ginning outturn character under 7 environment.It located under multiple environment 5 main effect QTL s (qBW-1-1, qBW-1-2, qLP-2-1, qLP-2-2 and qLP-4-2) of stable expression, detect 4 pairs of bell weights Upland cotton (Gossypium hirsutum under the multi-environments such as epistatic interaction QTLs and 7 couples of ginning outturn epistatic interaction QTLs Jia Fei L.) RIL bell weight and the qtl analysis Molecular Plant Breedings of ginning outturn character, 2011,9 (8):318-326)；Wang Tiankang etc. Utilize and be widely applied kind Shandong cotton with 2 fiber quality elite clone 0-153 and new land morning 24 and 2 and grind 28 and Ji cotton 516 For parent, the double cross F1 colonies of preparation, 4 SSR markers chain to 3 fibre strength main effect QTL s carry out assisted Selection, and Different Q TLs assembly effect is studied.As a result the selection of 4 marks all shows significant hereditary effect（Wang Tiankang Deng cotton fiber strength main effect QTL s molecular marker assisted selections and assembly effect research, Cotton Science, 2014,26 (5): 396-403)；Jamshed utilizes RIL（RIL）Informative population genetic map, navigates to 47 QTL under multiple environment Stable, these QTL are more to be existed in the form of coherent group, controls two or more characters, these QTL are concentrated mainly on 4, 7th, 14, No. 25 chromosome（Jamshed et al.Identification of stable quantitative trait loc (QTLs) for fiber quality traits across multiple environments in Gossypium hirsutum recombinant inbred line population.BMC Genomics,2016,17:197）；Zhang etc. The recombinant inbred lines that are built by two Upland Cottons 0-153 and SGK9708 have 5521 using SLAF sequence constructs Individual single nucleotide polymorphism, dense genetic map (the Zhang et that the total distance of covering is 3259.37cM al.Construction of a high-density genetic map by specific locus amplified fragment sequencing (SLAF-seq) and its application to Quantitative Trait Loci (QTL) analysis for boll weight in upland cotton (Gossypium hirsutum.).BMC Plant Biology,2016,16:79)。
In a word, SNP marker is current most potential molecular labeling, has been widely used at present, but in cotton Application in spending is also less, and majority is to utilize segregating population such as F in the research of forefathers2、BC1, genetic background complexity, or only in list Obtained result is detected under individual environment, therefore lacks reliability and stability, the stable QTL of multi-environment is few, and some researchs are most First purpose is intended merely to carry out the positioning of target gene.
The content of the invention
The technical problems to be solved by the invention are：By filtering out a kind of and upland cotton fiber rheobase because of close linkage And stable SNP marker is showed under multiple environment, these SNP markers are applied to the auxiliary of cotton fiber quality Selection, the fiber quality level of Cotton in China kind can be improved as early as possible.
The technical scheme that the present invention is provided is：A kind of SNP marker with upland cotton fiber high intensity gene linkage, is positioned at In 8 QTL related to intensity, wherein there is 4 to detect stable under multi-environment, these QTL are all located at No. 25 chromosomes, there is 3 Individual can screen obtains the preferable SNP marker of parting；Wherein withqFS-chr25-2Chain SNP marker is CRI- SNP198784、CRI-SNP198785、CRI-SNP198786、CRI-SNP198787、CRI-SNP198788、CRI- SNP198789、CRI-SNP198790、CRI-SNP198791；WithqFS-chr25-3Chain SNP marker is CRI- SNP198792、CRI-SNP198793、CRI-SNP198794、CRI-SNP198795；WithqFS-chr25-5Chain SNP marks Be designated as CRI-SNP198796, CRI-SNP198797, CRI-SNP198798, CRI-SNP198799, CRI-SNP198800, CRI-SNP198801、CRI-SNP198802、CRI-SNP198803、CRI-SNP198804、CRI-SNP198805、CRI- SNP198806, CRI-SNP198807,（Wherein CRI:Cotton Research Institute represent the Chinese Academy of Agricultural Sciences Cotton Research Institute；SNP represents type；Digitized representation marker development order）, described SNP marker is on chromosome Position and mutating alkali yl it is as shown in the table：
Mark title SNP site Mutating alkali yl
CRI-SNP198784 11634304 G/T
CRI-SNP198785 11727497 G/T
CRI-SNP198786 11727737 G/T
CRI-SNP198787 11800162 G/A
CRI-SNP198788 11800115 G/T
CRI-SNP198789 12039916 G/A
CRI-SNP198790 12039883 C/T
CRI-SNP198791 12364008 T/C
CRI-SNP198792 13627793 C/T
CRI-SNP198793 13795574 G/T
CRI-SNP198794 13627853 T/C
CRI-SNP198795 13795623 C/T
CRI-SNP198796 15620852 G/A
CRI-SNP198797 16378403 T/C
CRI-SNP198798 17002845 C/T
CRI-SNP198799 16735882 G/A
CRI-SNP198800 16727879 C/A
CRI-SNP198801 16548321 G/T
CRI-SNP198802 17481142 G/C
CRI-SNP198803 17882165 T/C
CRI-SNP198804 18212914 G/A
CRI-SNP198805 18213630 A/C
CRI-SNP198806 18213276 G/A
CRI-SNP198807 19698812 C/G
In the present invention, QTL names are with reference to McCouch etc.（1997）Naming rule in paddy rice, with q+ characters+linkage group+QTL The form of number is represented.（McCouch SR, Cho YG, Yano M, et al. Report on QTL nomenclature, Rice Genet Newslett.,1997,14:11-13）, for exampleqFS-chr25-2Expression navigates to 25 Number chromosome second QTL related to fibre strength.
SNP marker of the present invention can efficiently differentiate different SNP sites and different bases by the experiment of SNP Genotypings Because of type, so as to be screened to different cotton samples, the high strain of fibre strength can be filtered out, breeding cycle is greatly shortened, Improve the breeding efficiency of cotton fiber strength.
Meanwhile, the present invention also provides a kind of and upland cotton fiber intensity three QTLqFS-chr25-2、qFS-chr25- 3、qFS-chr25-5The screening technique of chain SNP marker, comprises the following steps：
（1）The choosing of hirsutum cultivar CCRI 41 promoted using crop field is SGK9708 and with Asiatic cotton high-intensity fiber base The excellent strain 0-153 of upland cotton of cause is that parent builds F2And F2:3Colony；
（2）F2:3Per generation selfing in colony's family, in F2:6A Single-plant selection is carried out from generation to generation, then plants for two generations to F6:8, F6:8And the later generation carries out multiple years experiment as recombinant inbred lines；
（3）Extract recombinant inbred lines and the DNA of parent；Specific method refers to documents below, and Song Guoli, modified CTAB method is fast Speed extracts cotton DNA, Cotton Science, 1998,10 (5):273-275)；
（4）Build linkage map：Digestion experiment is carried out to each sample genomic DNA of detection, to obtained endonuclease bamhi（SLAF Label）Carry out 3 ' end plus A processing, connect Dual-index sequence measuring joints, PCR amplifications, purifying, sample mixing, cut glue selection purpose piece Section, sequencing, the structure of genetic map is carried out to sequencing result with software（Zhang J, Guo W Z, Zhang T Z. Molecular linkage map of al-lotetraploid cotton (Gossypium hirsutum L. × Gossypium bar-badense L.) with a haploid population. Theor Appl Genet, 2002, 105: 1166–1174）；
（5）Fibre strength QTL is positioned：The stable fibre strength main effect QTL s screenings of multi-environment are carried out, be can obtain above-mentioned more than 3 The fibre strength main effect QTL s and its linked marker of ambient stable.
Beneficial effects of the present invention are as follows：
The site relevant with multi-environment is stable high-intensity fiber major gene resistance of the present invention has 3（qFS-chr25-2、qFS-chr25-3、qFS-chr25-5）, by screening with major gene of high-strength cotton fibre close linkage and under multiple environment Stable SNP marker is showed, these SNP markers are applied to the assisted Selection of cotton fiber quality, QTL positioning results are reliable, The fiber quality level of Cotton in China kind can be improved as early as possible.qFS-chr25-2Can be under 3 environment（(Anyang in 2007, Linqing, Anyang in 2010 in 2008）Detect, explainable phenotypic variation is 4.75-5.57%, additive effect value is 0.58- 0.71cN/tex；qFS-chr25-3Can be under 5 environment（Anyang, Linqing, Quzhou, peace in 2010 in 2009 in 2008 in 2007 Sun, Gaoyi in 2010）Detect, explainable phenotypic variation is 6.43-11.82%, additive effect value is 0.65-1.01cN/ tex；qFS-chr25-5Can be under 6 environment（Anyang in 2008, Anyang in 2009, Aksu in 2009, Gaoyi in 2010, Zhengzhou, Anyang in 2013 in 2010）Detect, the phenotypic variation of explanation is 4.88-14.60%, and additive effect value is 0.56- 1.34cN/tex.The present invention utilizes RIL F6:8（RIL）Filter out stable fibre strength QTLs and its close linkage Molecular labeling, the SNP marker is that the method for resurveying sequence by genome is obtained using the stable RIL colonies of cotton as material Arrive, the strain that fibre strength is improved is gone out using the molecular marker screening with these QTL close linkages, carry out molecular labeling auxiliary Breeding selection is helped, breeding cycle is substantially shorter, the breeding efficiency of cotton fiber strength is improved.
Brief description of the drawings
Fig. 1 is to resurvey total figure that sequence obtains away from the genetic map for 5197.17cM by genome.
Fig. 2 is QTLs chain with intensity on No. 25 chromosomes location drawing, and wherein multi-environment is stable and can screen SNP marker has 3, is respectivelyqFS-chr25-2、qFS-chr25-3、qFS-chr25-5。
Embodiment
Below by embodiment detailed description come the present invention is furture elucidated, but be not to the present invention limit System, is only illustrated.
（1）RIL F6:8Acquisition
The extraction of the field planting and DNA of -2008 years 2007 is see patent application publication number：CN 101613761A, invention name Claim：The patent application document of chain SSR marker with cotton fiber strength major gene resistance.2009 respectively at Earthquake of Anyang station in Henan China It is limited that Cotton Research Institute experiment station of Academy of Agricultural Sciences, Quzhou experiment station of China Agricultural University and the good science and technology of Aksu of Xinjiang's moral plant industry Plant parent and F in experiment station of company6:10Colony.Anyang and Quzhou use single file area, 5 meters of row length, line-spacing be respectively 0.8m and (0.8+0.5) m, often 20 plants of row；Xinjiang uses 6 row areas, and 2 meters of rows are grown, often 15 plants of row.2010 respectively at Gaoyi original seed , experiment station of the Earthquake of Anyang station in Henan Chinese Academy of Agricultural Sciences and Zhengzhou, henan plantation parent and F6:11Colony, Anyang and Zhengzhou are using single Row area, row long 5m, line-spacing 0.8m；Gaoyi uses single file area, long 4 m of row, wide-and narrow-row（0.8+0.6）m.Each above-mentioned pilot is all adopted Full RANDOMIZED BLOCK DESIGN is cannotd be used up, two repetitions are planted.Mid or late September carries out field sampling, and per stirpes receives flower, takes 12g or so Fiber sample carry out fiber quality measure.
（2）Extract recombinant inbred lines and the DNA of parent.
（3）The cotton tetraploid genome sequence provided with the Chinese Academy of Agriculture Science and Technologys Cotton Research Institute is selected to arrange as reference gene Group carries out electronics digestion prediction（Li FG, Fan GY, Lu CR, Xiao GH, Zou CS, Kohel RJ, Ma ZY, Shang HH, Ma XF, Wu JH, et al. Genome sequence of cultivated Upland cotton (Gossypium hirsutum TM-1) provides insights into genome evolution. Nature Biotechnology, 2015, 33(5)）, final choice HaeIII+SspI enzymes, digestion mark rate is 98.61%, is obtained 495.48Mreads, endonuclease bamhi length is SLAF labels in 364-414bp sequence definition.
（4）According to selected most suitable digestion scheme, digestion experiment is carried out to each sample genomic DNA of detection, to obtaining Endonuclease bamhi（SLAF labels）Carry out 3 ' end plus A processing, connection Dual-index be sequenced street corner, PCR amplifications, purifying, sample mixing, Cut glue and choose purpose fragment, be sequenced after library quality inspection is qualified with IlluminaHiseqTM2500.
（5）It is identified using the Dual-index initial data obtained to sequencing, obtains the reads of each sample.It is logical The method clustered between reads is crossed, SLAF labels are developed in parent and filial generation.
（6）By bioinformatic analysis, 321797 SLAF labels are obtained, the wherein SLAF labels of polymorphism have 35300.
（7）Genotype codes are carried out to the SLAF labels of polymorphism, genotype codes rule is 2 general equipotentials of science of heredity Coding rule, such as parent genotype of certain mark are aa（Male parent）And bb（It is maternal）, progeny genotypes ab then represents that the sample exists The type of coding of this mark is heterozygosis, wherein there is a genotype to come from male parent, has a genotype to come from female parent.
（8）To ensure genetic map quality, SLAF labels are sequenced below depth 10x, integrity degree according to Parent and are less than 30%th, the serious (p-value of separation partially<0.05), parent hybrid, filtered while comparing to the conditions of two sets genomes, it is common to sieve The 7958 SLAF labels selected.
（9）SLAF labels are divided into by 26 linkage groups by the positioning with reference gene group, high-quality molecular label is calculated Between LOD value, chain point of group is carried out by LOD value, to each linkage group using HighMap softwares carry out genetic map structure Build, by correction, obtain total figure away from the genetic map for 5197.17cM（As shown in Figure 1）.Wherein HighMap softwares are by Beijing The independent research of hundred mikeys biology scientific ＆ technical corporation.
（10）Sequencing data based on SLAF, is compared by BWA and the reference gene group of two 2 times of body cottons, obtained and parent Between have polymorphism SNP have 44583, after mass filter, on collection of illustrative plates positioning obtain 10440 SNP markers.
（11）Using software QTL IciMapping V4 .0 (http://www .isbreeding .net/ Software/) and software WinQTLCart 2.5, (Anyang in 2007, Anyang in 2008, faced within 2008 by 11 environment Clearly, Quzhou in 2008, Anyang in 2009, Quzhou in 2009, Aksu in 2009, Anyang in 2010, Gaoyi in 2010,2010 Zhengzhou, Anyang in 2013）The phenotypic data and genotype data of fibre strength character, carry out the multi-environment of fibre strength character QTL positioning analysises, are obtained the QTL related to intensity totally 5, wherein multi-environment it is stable and can screen SNP marker have 3 （As shown in Figure 2）, wherein withqFS-chr25-2Chain SNP marker is CRI-SNP198784, CRI-SNP198785, CRI- SNP198786、CRI-SNP198787、CRI-SNP198788、CRI-SNP198789、CRI-SNP198790、CRI- SNP198791；WithqFS-chr25-3Chain SNP marker is CRI-SNP198792, CRI-SNP198793, CRI- SNP198794、CRI-SNP198795；WithqFS-chr25-5Chain SNP marker is CRI-SNP198796, CRI- SNP198797、CRI-SNP198798、CRI-SNP198799、CRI-SNP198800、CRI-SNP198801、CRI- SNP198802、CRI-SNP198803、CRI-SNP198804、CRI-SNP198805、CRI-SNP198806、CRI- SNP198807。

Claims (4)

1. a kind of and cotton fiber high intensity gene linkage SNP marker, it is positioned at 5 related to fibre strength In QTL, wherein there is 4 to detect stable under multi-environment, these QTL are all located on No. 25 chromosomes of upland cotton, have 3 can sieve Choosing obtains the preferable SNP marker of parting, wherein withqFS-chr25-2Chain SNP marker is CRI-SNP198784, CRI- SNP198785、CRI-SNP198786、CRI-SNP198787、CRI-SNP198788、CRI-SNP198789、CRI- SNP198790、CRI-SNP198791；WithqFS-chr25-3Chain SNP marker is CRI-SNP198792, CRI- SNP198793、CRI-SNP198794、CRI-SNP198795；WithqFS-chr25-5Chain SNP marker is CRI- SNP198796、CRI-SNP198797、CRI-SNP198798、CRI-SNP198799、CRI-SNP198800、CRI- SNP198801、CRI-SNP198802、CRI-SNP198803、CRI-SNP198804、CRI-SNP198805、CRI- SNP198806, CRI-SNP198807, it is characterised in that：The position of described SNP marker on chromosome and mutation alkali Base is as shown in the table：
Mark title SNP site Mutating alkali yl CRI-SNP198784 11634304 G/T CRI-SNP198785 11727497 G/T CRI-SNP198786 11727737 G/T CRI-SNP198787 11800162 G/A CRI-SNP198788 11800115 G/T CRI-SNP198789 12039916 G/A CRI-SNP198790 12039883 C/T CRI-SNP198791 12364008 T/C CRI-SNP198792 13627793 C/T CRI-SNP198793 13795574 G/T CRI-SNP198794 13627853 T/C CRI-SNP198795 13795623 C/T CRI-SNP198796 15620852 G/A CRI-SNP198797 16378403 T/C CRI-SNP198798 17002845 C/T CRI-SNP198799 16735882 G/A CRI-SNP198800 16727879 C/A CRI-SNP198801 16548321 G/T CRI-SNP198802 17481142 G/C CRI-SNP198803 17882165 T/C CRI-SNP198804 18212914 G/A CRI-SNP198805 18213630 A/C CRI-SNP198806 18213276 G/A CRI-SNP198807 19698812 C/G
。
2. a kind of screening technique of the SNP marker described in claim 1, comprises the following steps：
（1）The choosing of hirsutum cultivar CCRI 41 promoted using crop field is SGK9708 and with Asiatic cotton high-intensity fiber base The excellent strain 0-153 of upland cotton of cause is that parent builds F2And F2:3Colony；
（2）F2:3Per generation selfing in colony's family, in F2:6A Single-plant selection is carried out from generation to generation, then plants for two generations to F6:8, F6:8And the later generation carries out multiple years experiment as recombinant inbred lines；
（3）Extract recombinant inbred lines and the DNA of parent；
（4）Build linkage map：Digestion experiment is carried out to each sample genomic DNA of detection, to obtained endonuclease bamhi（SLAF Label）Carry out 3 ' end plus A processing, connect Dual-index sequence measuring joints, PCR amplifications, purifying, sample mixing, cut glue selection purpose piece Section, sequencing carries out the structure of genetic map to result；
（5）Fibre strength QTL is positioned：Fibre strength main effect QTL s screenings stable under multiple environment are carried out, are selected strong with fiber The obvious SNP of parting is spent, the fibre strength master of multiple ambient stables of 3 described in claim 1 from upland cotton is obtained Imitate QTLsqFS-chr25-2、qFS-chr25-3、qFS-chr25-5And its chain SNP marker.
3. the SNP marker described in claim 1 can be tested by the Genotyping of SNP marker, so as to efficiently differentiate not With SNP site and different genotype, different cotton samples can be screened, reach the purpose of molecular mark.
4. application according to claim 3, can filter out the high strain of fibre strength, greatly shorten breeding cycle, improve The breeding efficiency of cotton fiber strength.
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