CN106755432A - One upland cotton chromosome segment that can dramatically increase sea island cotton ginning outturn and its SSR label primer and application - Google Patents
One upland cotton chromosome segment that can dramatically increase sea island cotton ginning outturn and its SSR label primer and application Download PDFInfo
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Abstract
The invention discloses a upland cotton chromosome segment that can dramatically increase sea island cotton ginning outturn and its SSR label primer and application, the fragment is named as IL A13 5, comprising 5 SSR signatures：NAU3017245,SHIN‑1462230,JESPR204170,NAU5345160,BNL2571280；The genomic DNAs of Upland Cotton TM 1 are expanded using 5 described SSR label primers, the chromosome segment simultaneously containing 5 molecular labeling sites is upland cotton chromosome segment A13 5.The upland cotton chromosome segment of A13 5 is applied to Molecular breeding in upland cotton, can significantly increase sea island cotton ginning outturn.
Description
Technical field
The invention belongs to field of crop genetic breeding, it is related to the upland cotton chromosome that can dramatically increase sea island cotton ginning outturn
Fragment and its SSR label primer and application.
Technical background
Cotton is global Important Economic crop, is the important materials of textile industry.China is also Chan Mian big countries, cotton growing area
Covering 16 provinces, city (autonomous regions), more than 5,000,000 tons of annual total output.Lint yield main composition factor is that bell is weighed and ginning outturn,
Wherein ginning outturn influences huge to lint yield.
Ginning outturn is influenceed larger by quality hereditary capacity, also receives environmental influence.Domestic and international researcher is to fiber yield
Substantial amounts of QTL Position Research is carried out.Yin Jianmei etc. (2002) screens 4 ginning outturn QTL, explains the 5.9%- of phenotypic variation
25%；Wuetal (2003) detects 1 ginning outturn QTL, explains the 17.2% of phenotypic variation；Zhang Peitong etc. (2005) detects 2
Individual ginning outturn QTL, explains the 22.5% and 29.3% of phenotypic variation；Ten thousand groups (2007) are produced using Upland Cotton intermolecular hybrid
RIL colonies detect 5 ginning outturn QTL in 4 environment, can explain the phenotypic variation of 4.8%-63.8%；Field petrel
(2007) using Upland Cotton intermolecular hybrid F2Colony, detects 1 ginning outturn QTL, explains the 76.93% of ginning outturn variation；Qin Hong
Moral (2007) hands over combine detection to 2 ginning outturn QTL using between Upland Cotton four；Wang etc. (2007) is using between Upland Cotton
The recombinant inbred lines that hybridization is produced detect 4 ginning outturn QTL in the average value under 3 environment, explain 3.88%-
13.04% phenotypic variation；Wang etc. (2013) is detected using sea island cotton Luyuan 343 and upland cotton lumianyan 22
The related QTLs of 24 ginning outturn；Shi etc. (2015) detects 26 QTLs of ginning outturn, wherein 50% QTLs can improve material
The 1.07-2.41% of ginning outturn.
Sea island cotton breeding time is long, and ripe evening, bell is small, and yield is less than upland cotton, but fiber finer is strong, is high-count yarn spun raw material,
Therefore, the yield traits of sea island cotton is improved, it is significant in breeding.Traditional crossbreeding is separated sternly because of extra large land F2 generations
It is difficult to carry out again, therefore, carry out sea island cotton yield traits important means of the improvement as sea island cotton breeding using introgressive line.
Chromosome segment introgressive line (Chromosome segment introgression lines, CSIL) is using miscellaneous
Hand over, the whole base of nurse crop that backcrossing and molecular marker assisted selection (marker-assisted selection, MAS) build
Because of a series of NILs organized.A chromosome segment for homozygosis in its genome only from donor parents, and
The remainder of genome is identical with recurrent parent.It is to carry out genome research, and particularly QTL is positioned and Molecular design breeding
Ideal material.Eshed and Zamir (1995) located using 50 chromosome segment introgressive lines for covering tomato full-length genome
The QTL of individual Quality Characteristics in Tomato.They are had found using chromosome segment introgressive line to tamato fruit total solids content and fruit
Weight carries out Billy when QTL is positioned and to have more 1 times with the QTL numbers that conventional segregating population is positioned；Rae et al. (1999) are grinding
Found when studying carefully crucifer florescence QTL, the QTL numbers Billy identified using introgressive line uses what conventional segregating population was identified
QTL numbers are more.Arbelaez etc. (2015) constructs introgressive line using two wild species rice and a common recurrent parent.In addition
In soybean (Wang etc. 2013；2014), corn (Burton etc. 2015) has related report to Lee etc. in ramie (Liu etc. 2014)
Road.
Wang etc. (2012) constructs 105 Introgressed lines using upland cotton and sea island cotton, is reflected with 278 polymorphism marks
It is fixed, introgressed segment length 333.5cM, the 6.7% of covering coloring body fragment.To each proterties of Introgressed line material and polymorphism mark
Between be associated analysis, there are 40 pairs of SSR markers to be associated with 5 fiber quality correlated traits, average contribution rate：6.31%.Wang
Constructed with upland cotton (TM-1) as background Deng (2012), sea island cotton (H7124) is the Introgressed line of donor parents, has 169
Family, single slice introgressive line has 51, accounts for 30.18%, in BC523 conspicuousness marks, wherein fabric are detected in colony
17 of matter, Yield Components (Single boll weight and ginning outturn) 6.In BC5S127 conspicuousnesses mark, wherein fabric are detected in colony
22 of matter, Yield Components (Single boll weight and ginning outturn) 5.14 conspicuousness marks, wherein fiber are detected in CSIL colonies
12 of quality, Yield Components (Single boll weight and ginning outturn) 2.
Cao etc. (2014) has utilized the dyeing built by upland cotton TM-1 (recurrent parent) and extra large 7124 (nonrecurrent parents)
Body fragment substitution line has carried out the practice of molecular breeding, using 4 introgressive lines containing the excellent fiber quality QTLs of sea island cotton
It is (new that (IL019-D6-1, IL019-A2-6, IL088-A7-3 and IL040-A4-1) improves four Xinjiang precocity cotton variety (being)
Land early 26, new land early 41, new land early 42 and fiber quality 0768).
The content of the invention
It is an object of the invention to provide the upland cotton chromosome segment that can dramatically increase sea island cotton ginning outturn.
It is a further object of the present invention to provide the molecular labeling and its SSR primers of upland cotton chromosome segment A13-5.
The purpose of the present invention can be achieved through the following technical solutions：
One upland cotton chromosome segment A13-5 that can significantly improve island cotton boll weight, this upland cotton chromosome segment
A13-5 comes from Upland Cotton TM-1, comprising 5 SSR signatures：NAU3017245,SHIN-1462230,
JESPR204170,NAU5345160,BNL2571280；Upland Cotton TM-1 genes are expanded using 5 described SSR label primers
Group DNA, the chromosome segment simultaneously containing 5 molecular labeling sites is upland cotton chromosome segment A13-5；Described 5
SSR label primer and the purpose fragment length of amplification are as follows：
NAU3017245Forward primer sequence be SEQ ID NO.1, reverse primer sequences be SEQ ID NO.2, on land
In cotton variety TM-1 it is amplifiable go out length for 245bp DNA fragmentation；
SHIN-1462230Forward primer sequence be SEQ ID NO.3, reverse primer sequences be SEQ ID NO.4, in land
In ground cotton variety TM-1 it is amplifiable go out length be the DNA fragmentation of 250bp；
JESPR204170Forward primer sequence be SEQ ID NO.5, reverse primer sequences be SEQ ID NO.6, in land
In ground cotton variety TM-1 it is amplifiable go out length be the DNA fragmentation of 170bp；
NAU5345160Forward primer sequence be SEQ ID NO.7, reverse primer sequences be SEQ ID NO.8, on land
In cotton variety TM-1 it is amplifiable go out length for 160bp DNA fragmentation.
BNL2571280Forward primer sequence be SEQ ID NO.9, reverse primer sequences be SEQ ID NO.10, in land
In ground cotton variety TM-1 it is amplifiable go out length be the DNA fragmentation of 280bp.
Applications of the described upland cotton chromosome segment A13-5 in fiber yield breeding is improved.
The side of the upland cotton introgressive line IL-A13-5 that ginning outturn is significantly improved is obtained using chromosome segment A13-5 of the present invention
Method, comprises the steps of：
(1) new 25 (♂) of sea of island cotton variety and upland cotton Genetic standard line TM-1 (♀) hybridization produces F1 generation, then with new
Sea 25 is recurrent parent, is returned 3 times, 4 generation BC3S4 seeds of selfing.
(2) BC is planted3S4Seed produces plant, DNA is extracted using CTAB methods, using molecular labeling primer NAU3017245,
SHIN-1462230,JESPR204170,NAU5345160,BNL2571280DNA is entered performing PCR amplification, the primer of each molecular labeling and
The purpose fragment length of amplification is as follows：
NAU3017245Forward primer sequence be SEQ ID NO.1, reverse primer sequences be SEQ ID NO.2, on land
In cotton variety TM-1 it is amplifiable go out length for 245bp DNA fragmentation；
SHIN-1462230Forward primer sequence be SEQ ID NO.3, reverse primer sequences be SEQ ID NO.4, in land
In ground cotton variety TM-1 it is amplifiable go out length be the DNA fragmentation of 250bp；
JESPR204170Forward primer sequence be SEQ ID NO.5, reverse primer sequences be SEQ ID NO.6, in land
In ground cotton variety TM-1 it is amplifiable go out length be the DNA fragmentation of 170bp；
NAU5345160Forward primer sequence be SEQ ID NO.7, reverse primer sequences be SEQ ID NO.8, on land
In cotton variety TM-1 it is amplifiable go out length for 160bp DNA fragmentation.
BNL2571280Forward primer sequence be SEQ ID NO.9, reverse primer sequences be SEQ ID NO.10, in land
In ground cotton variety TM-1 it is amplifiable go out length be the DNA fragmentation of 280bp.
Containing 5 molecular labeling sites, (molecular weight is respectively 245bp simultaneously for selection；250bp；170bp；160bp；
Chromosome segment 280bp) is sea island cotton chromosome segment A13-5；
(3) by the introgressive line IL-A13-5 containing the chromosome segment, Xinjiang was planted in respectively at 2013 and 2014
Aksu Prefecture and Korla City proving ground, plant two rows, and row is long 2 meters, and film is wide 0.75 meter, three repetitions, while planting TM-
1 and Xin Hai 25, treats that cotton collects 25 cotton bolls of every row material, weighs, and then cotton ginning claims gined cotton weight, calculates ginning outturn.By
The data drawn after actual survey calculation are：2 years ginning outturn of introgressive line IL-A13-5 containing upland cotton chromosome segment A13-5
Average value is respectively 39.31% and 37.43%, than compareing new by extra large 25 increase by 7.52% and 10.54% respectively.
Molecular labeling primer of the present invention is screening upland cotton chromosome segment A13-5 to improve fiber yield breeding
In application.
Described 5 SSR label primers amplification Upland Cotton TM-1 genomic DNAs are preferably used, 5 points are contained simultaneously
The as upland cotton chromosome segment A1-6 of the purpose fragment of son mark amplification, selection contains upland cotton chromosome segment A13-5
Material reserve seed for planting.
A kind of method for screening upland cotton chromosome segment A13-5, including the 5 SSR label primers amplification described in use
Upland Cotton TM-1 genomic DNAs, the chromosome segment simultaneously containing 5 purpose fragments of molecular labeling amplification is land
Cotton chromosome fragment A13-5.
Beneficial effect
1st, the one provided by the present invention upland cotton chromosome segment A13-5 that can dramatically increase sea island cotton ginning outturn, and will
It is applied to breeding and obtains a upland cotton introgressive line IL-A13-5 that can dramatically increase sea island cotton ginning outturn, by 2 years 2 points
After 2 repetitions are tested, it is found that the chromosome segment can dramatically increase sea island cotton ginning outturn, 2 points of extra large 25 increasings newer than control respectively in 2 years
Plus 7.52% and 10.54% (table 2).Therefore the sea island cotton chromosome segment in IL-A13-5 chromosome segment introgressive lines can be significantly
Increase sea island cotton ginning outturn, it is significant in sea island cotton breeding.
2nd, 5 SSR molecular markers and its primer of sea island cotton chromosome segment A13-5 provided by the present invention can increase sea
The efficiency of selection (Fig. 1) of island cotton dress point.Molecular labeling auxiliary mark Piece Selection, can accelerate rearing new variety process.
Brief description of the drawings
Amplification site of the 5 SSR primers of Fig. 1 in new sea 25, TM-1 and IL-A13-5
Specific embodiment
Embodiment 1
Agricultural University Of Nanjing's Cotton Research Institute with the new sea 25 of sea island cotton as recurrent parent, upland cotton Genetic standard line TM-1 product
It is donor parents (nonrecurrent parent) to plant, and is returned 3 times, 4 generation BC of selfing3S4Seed.New 25 and TM-1 of sea is big through Nanjing agriculture
After learning Cotton Research Institute introduction, selfing for many years preserves breeding, if other colleagues need, Agricultural University Of Nanjing's Cotton Research Institute can
These germplasm are provided to the public.
Plantation BC3S4Plant, every plant of not deployed blade of collection 3-4 pieces is put into the centrifuge tube of 1.5ml, and adds precooling
Fresh the μ l of Extraction buffer 600, with beveller grind, using CTAB methods (Paterson A H, Brubaker C
L,Wendel J F.A rapid method for extraction of cotton(Gossypium spp.)genomic
DNA suitable for RFLP or PCR analysis[J].Plant Molecular Biology Reporter,
1993,11(2):DNA 122-127.) is extracted, is carried out with SSR primers BNL2440, NAU2437, NAU3901, dPL0526 (table 1)
PCR is expanded, the μ l of amplification system 10, DNA profiling 1 μ l, 94 DEG C of predegenerations 5min, 94 DEG C of denaturation 0.5min, 57 DEG C of renaturation 0.5min,
72 DEG C of extension 1min, after 30 circulations, 72 DEG C re-extend 10min, and amplified production is through native polyacrylamide gel electrophoresis：It is solidifying
Gum concentration is 8%, and electrophoretic buffer is 0.5 times of TBE, 220V constant pressures electrophoresis 1 hour.Selection contains 5 molecular labeling bars simultaneously
Band (is respectively 245bp in TM-1 middle-molecular-weihydroxyethyls；250bp；170bp；160bp；Chromosome segment 280bp) is land cotton
Chromosome fragment A13-5.
SSR molecular marker on the A13-5 chromosome segments of table 1
Note:BNL numbering primer from Research Genetics companies of the U.S. (http://www.resgen.com)；
The primer of NAU numberings developed from EST-SSR sequences by this laboratory (Zhao L, Yuanda L, Caiping C,
et al.Toward allotetraploid cotton genome assembly:integration of a high-
density molecular genetic linkage map with DNA sequence information[J].BMC
genomics,2012,13(1):539；The primer of JESPR numberings is carried by Brian Scheffler and Jodi Scheffler
For；The primer of SHIN numberings comes from Monsanto companies of the U.S. (http://www.monsanto.com).All of primer information
Can be downloaded from the www.cottonmarker.org of website..
By the introgressive line IL-A13-5 containing the chromosome segment, Xinjiang Acker was planted in respectively at 2013 and 2014
Soviet Union area and Korla City proving ground, plant two rows, and row is long 2 meters, and film is wide 0.75 meter, secondary repetition, at the same plant TM-1 and
New sea 25, treats that cotton collects 25 cotton bolls of every row material, weighs, and cotton ginning weighs gined cotton weight, calculates ginning outturn.By actual survey
Measuring the data drawn after calculating is：2 years ginning outturn average values of introgressive line IL-A13-5 containing upland cotton chromosome segment A13-5
Respectively 39.31% and 37.43%, than compareing new by extra large 25 increase by 7.52% and 10.54% respectively, amplification is notable.Therefore A13-5
Upland cotton chromosome segment be applied to Molecular breeding in upland cotton, sea island cotton ginning outturn can be significantly increased.
The introgressive line A13-5 of table 2 ginning outturn performances in different year, varying environment
SEQUENCE LISTING
<110>Agricultural University Of Nanjing
<120>One upland cotton chromosome segment that can dramatically increase sea island cotton ginning outturn and its SSR label primer and application
<160> 10
<210> 1
<211> 20
<212> DNA
<213>Artificial sequence
<213>NAU3017 forward primers
<400> 1
cacccaaacc ctaattcaac 20
<210> 2
<211> 20
<212> DNA
<213>Artificial sequence
<213>NAU3017 is reversely to primer
<400> 2
gcatcttcaa gggaagaaaa 20
<210> 3
<211> 20
<212> DNA
<213>Artificial sequence
<213>SHIN-1462 forward primers
<400> 3
ctagaaagct cgtgggatgg 20
<210> 4
<211> 23
<212> DNA
<213>Artificial sequence
<213>SHIN-1462 reverse primers
<400> 4
gcacagatga cggatattac aga 23
<210> 5
<211> 19
<212> DNA
<213>Artificial sequence
<213>JESPR204 forward primers
<400> 5
ctccaggttc aatggtctg 19
<210> 6
<211> 20
<212> DNA
<213>Artificial sequence
<213>JESPR204 reverse primers
<400> 6
gccatgttgg acaagtagtc 20
<210> 7
<211> 20
<212> DNA
<213>Artificial sequence
<213>NAU5345 forward primers
<400> 7
aaaggcagag aaacctgatg 20
<210> 8
<211> 20
<212> DNA
<213>Artificial sequence
<213>NAU5345 reverse primers
<400> 8
cctccaaagg taaagcaaga 20
<210> 9
<211> 20
<212> DNA
<213>Artificial sequence
<213>BNL2571 forward primers
<400> 9
tcgctatcgc tctgaaatca 20
<210> 10
<211> 20
<212> DNA
<213>Artificial sequence
<213>BNL2571 reverse primers
<400> 10
atgccacgga attagcaaac 20
Claims (6)
1. a upland cotton chromosome segment A13-5 for dramatically increasing fiber yield, it is characterised in that come from upland cotton TM-1,
Positioned at the A13 chromosomes of cotton chromosome group, comprising 5 SSR signatures：NAU3017245,SHIN-1462230,
JESPR204170,NAU5345160,BNL2571280；Upland Cotton TM-1 genes are expanded using 5 described SSR label primers
Group DNA, the chromosome segment simultaneously containing 5 molecular labeling sites is upland cotton chromosome segment A13-5；Described 5
SSR label primer and the purpose fragment length of amplification are as follows：
NAU3017245Forward primer sequence be SEQ ID NO.1, reverse primer sequences be SEQ ID NO.2, in upland cotton product
In kind of TM-1 it is amplifiable go out length be the DNA fragmentation of 245bp；
SHIN-1462230Forward primer sequence be SEQ ID NO.3, reverse primer sequences be SEQ ID NO.4, in upland cotton
In kind TM-1 it is amplifiable go out length for 250bp DNA fragmentation；
JESPR204170Forward primer sequence be SEQ ID NO.5, reverse primer sequences be SEQ ID NO.6, in upland cotton
In kind TM-1 it is amplifiable go out length for 170bp DNA fragmentation；
NAU5345160Forward primer sequence be SEQ ID NO.7, reverse primer sequences be SEQ ID NO.8, in upland cotton product
In kind of TM-1 it is amplifiable go out length be the DNA fragmentation of 160bp；
BNL2571280Forward primer sequence be SEQ ID NO.9, reverse primer sequences be SEQ ID NO.10；In upland cotton
In kind TM-1 it is amplifiable go out length for 280bp DNA fragmentation.
2. 5 SSR molecular marker primers of the sea island cotton chromosome segment A13-5 described in claim 1, it is characterised in that：5
SSR marker is respectively NAU3017245,SHIN-1462230,JESPR204170,NAU5345160,BNL2571280；Each molecular labeling
Primer and amplification purpose fragment length it is as follows：
NAU3017245Forward primer sequence be SEQ ID NO.1, reverse primer sequences be SEQ ID NO.2, in upland cotton product
In kind of TM-1 it is amplifiable go out length be the DNA fragmentation of 245bp；
SHIN-1462230Forward primer sequence be SEQ ID NO.3, reverse primer sequences be SEQ ID NO.4, in upland cotton
In kind TM-1 it is amplifiable go out length for 250bp DNA fragmentation；
JESPR204170Forward primer sequence be SEQ ID NO.5, reverse primer sequences be SEQ ID NO.6, in upland cotton
In kind TM-1 it is amplifiable go out length for 170bp DNA fragmentation；
NAU5345160Forward primer sequence be SEQ ID NO.7, reverse primer sequences be SEQ ID NO.8, in upland cotton product
In kind of TM-1 it is amplifiable go out length be the DNA fragmentation of 160bp.
BNL2571280Forward primer sequence be SEQ ID NO.9, reverse primer sequences be SEQ ID NO.10, in upland cotton
In kind TM-1 it is amplifiable go out length for 280bp DNA fragmentation.
3. applications of the upland cotton chromosome segment A13-5 described in claim 1 in fiber yield breeding is improved.
4. the molecular labeling primer described in claim 2 is educated in screening upland cotton chromosome segment A13-5 with improving fiber yield
Application in kind.
5. application according to claim 4, it is characterised in that the 5 SSR label primers amplification described in usage right requirement 2
Upland Cotton TM-1 genomic DNAs, contain 5 as upland cotton chromosome pieces of the purpose fragment of molecular labeling amplification simultaneously
Section A1-6, material of the selection containing upland cotton chromosome segment A13-5 is reserved seed for planting.
6. a kind of method for screening upland cotton chromosome segment A13-5, it is characterised in that including 5 described in usage right requirement 2
Individual SSR label primer expands Upland Cotton TM-1 genomic DNAs, the purpose fragment for being expanded containing 5 molecular labelings simultaneously
Chromosome segment is upland cotton chromosome segment A13-5.
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Cited By (4)
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|---|---|---|---|---|
| CN108359741A (en) * | 2018-05-22 | 2018-08-03 | 山东棉花研究中心 | The one InDel molecular labeling and its application with the 15th chromosome ginning outturn main effect QTL compact linkage of upland cotton |
| CN110093442A (en) * | 2019-05-09 | 2019-08-06 | 江苏省农业科学院 | SSR molecular marker relevant to the short bar of cotton and high ginning outturn |
| CN110129475A (en) * | 2019-05-09 | 2019-08-16 | 江苏省农业科学院 | The relevant SSR nucleic acid sequence of the high ginning outturn of cotton and its application |
| WO2020224636A1 (en) * | 2019-05-09 | 2020-11-12 | 江苏省农业科学院 | Gossypium anomalum-sourced ssr sequence associated with high lint percentage and drought tolerance and application thereof |
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Cited By (5)
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|---|---|---|---|---|
| CN108359741A (en) * | 2018-05-22 | 2018-08-03 | 山东棉花研究中心 | The one InDel molecular labeling and its application with the 15th chromosome ginning outturn main effect QTL compact linkage of upland cotton |
| CN108359741B (en) * | 2018-05-22 | 2021-07-27 | 山东棉花研究中心 | InDel molecular marker closely linked with major QTL (quantitative trait locus) of chromosome 15 of upland cotton and application thereof |
| CN110093442A (en) * | 2019-05-09 | 2019-08-06 | 江苏省农业科学院 | SSR molecular marker relevant to the short bar of cotton and high ginning outturn |
| CN110129475A (en) * | 2019-05-09 | 2019-08-16 | 江苏省农业科学院 | The relevant SSR nucleic acid sequence of the high ginning outturn of cotton and its application |
| WO2020224636A1 (en) * | 2019-05-09 | 2020-11-12 | 江苏省农业科学院 | Gossypium anomalum-sourced ssr sequence associated with high lint percentage and drought tolerance and application thereof |
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