CN114292942B - Main effect QTL for regulating and controlling corn leaf senescence, molecular marker and application thereof - Google Patents
Main effect QTL for regulating and controlling corn leaf senescence, molecular marker and application thereof Download PDFInfo
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Abstract
The invention relates to a major QTL for regulating and controlling corn leaf senescence, and a molecular marker and application thereof, belonging to the field of molecular genetic breeding. The main effective sites Stg3 and Stg7 have the function of regulating and controlling the senescence of corn leaves, are respectively positioned on the No.3 chromosome and the No.7 chromosome of corn, and have molecular markers which are respectively M36, A-3-11, A-7-6 and A-7-7. The molecular marker has important application value in screening and identifying corn resources with leaf senescence delay characteristics and finely positioning target genes in QTL, and provides important candidate sites for prolonging the photosynthesis functional period of crop leaves in molecular genetic breeding.
Description
The application is a divisional application with the application number of 2020111152910 and the invention name of ' two main effect QTLs for regulating and controlling corn leaf senescence ' and molecular markers and application thereof '.
Technical Field
The invention relates to a major QTL for regulating and controlling corn leaf senescence, and a molecular marker and application thereof, belonging to the field of molecular genetic breeding.
Background
Senescence is an important process of nutrient transfer in the final stages of leaf development, playing a vital role in ensuring crop yield. Initiation of leaf senescence is associated with leaf age (jin, H.C., hille, J., dijkwel, P.P.), ageing in plants conserved strategies and novel pathway.plant Biology,2003,5,455-464), in addition to which various biotic or abiotic stresses, such as pathogenic bacterial infection, drought stress and darkness stress, can also induce leaf senescence (Guo, Y., gan, S.S., convergence and divergence in gene expression profiles induced by leaf senescence and, 27 senesce-promoting hormonal, pathological and environmental stress events.plant Cell and Environment,2012,35,644-655). This process is accompanied by a number of physiological and biochemical and molecular level changes, such as chlorophyll degradation, disruption of the photosynthetic membrane system, changes in the relationship of the source bank and expression of thousands of age-related genes (SAGs). Delaying leaf senescence in good time is an effective means for prolonging leaf functional period and improving crop yield. It has been found that delaying leaf senescence can yield average corn at 0.29 tons/hectare (Zhang j., fengler k.a., van Hemert j.l., gupta, r., mongar, n., sun, j., allen, w.b., wang, y., weers, b., mo, h., lafitte, r., hou, z., bryant, a., ibraheme, f., arp, j., swaminothan, k, moose, s.p., li, b., shen, b., identification and characterization of a novel stay-green QTL that increases yield in mail ze.plant Biotechnology Journal,2019,17,2272-2285), average rice yield 10% (Mao, c, lu, s, lv, b, zhang, b, shen, j, he, j, luo, l, xi, d, chen, x, min, f, A rice NAC transcription factor promotes leaf senescence via ababiosynthesis.plant Physiology,2017,174,1747-1763), average wheat yield 6-28% (Christopher, j.t., manschadi, a.m., hammer, g.l., borrell, a.k., developmental and physiological traits associated with high yield and stay-green phenotype in wshellian Journal of Agricultural Research,2008,59,354-364). Therefore, elucidation of the molecular mechanism of leaf senescence is of great importance for crop breeding.
Corn (Zea Mays) is a major food crop in our country and is also an important industrial raw material and feed source. Frequent occurrence in extreme weather induces premature senility or death of the leaves, resulting in yield reduction or even absolute yield. Therefore, cultivation of the corn germplasm with delayed senescence is an important guarantee of stable yield. The development of sequencing technology and molecular marker assisted breeding technology lays a foundation for deep excavation of major QTL for controlling corn leaf senescence and application thereof.
To date, researchers have mapped 268 leaf senescence-associated on 10 chromosomes of maize by constructing multiple types of genetic populations (recombinant inbred, backcrossed, and doubled haploid populations, etc.) and developing multiple molecular markersMajor QTL, which are capable of interpreting 0.47-24.32% of leaf senescence phenotype variation (Almeida, g.d., nair, s., borem, a, cairns, j, tranchel, s, ribaut, j.m., banzier, m., prasanna, b.m., cross, j, babu, r., molecular mapping across three populations reveals a QTL hotspot region on chromosome 3for secondary traits associated with drought tolerance in tropical maize.Molecular breeding,2014,34,701-715; beavis, w.d., smith, o.s., grant, d., fincher, r., identification of quantitative trait loci using a small sample of topcrossed and F) 4 progeny from maize.Crop Science,1994,34,882-896；Belícuas,P.R.,Aguiar,A.M.,Bento,D.A.V.,T.M.M.,de Souza Junior,C.L.,Inheritance of the stay-green trait in tropical maize.Euphytica,2014,198,163-173；Khanal,R.,Navabi,A.,Lukens,L.,Linkage map construction and quantitative trait loci(QTL)mapping using intermated vs.selfed recombinant inbred maize line(Zea mays L.).Canadian Journal of Plant Science,2015,95,1133-1144；Messmer,R.,Fracheboud,Y.,/>M.,Stamp,P.,Ribaut,J.M.,Drought stress and tropical maize:QTLs for leaf greenness,plant senescence,and root capacitance.Field Crops Research,2011,124,93-103；Trachsel,S.,Sun,D.,SanVicente,F.M.,Zheng,H.,Atlin,G.N.,Suarez,E.A.,Babu,R.,Zhang,X.,Identification of QTL for early vigor and stay-green conferringtolerance to drought in two connected advanced backcross populations in tropical maize(Zea mays L.).PLoS One,2016,11,e0149636；Wang,A.,Li,Y.,Zhang,C.,QTL mapping for stay-green inmaize(Zea mays).Canadian Journal of Plant Science,2012,92,249-256；Yang,Z.,Li,X.,Zhang,N.,Wang,X.,Zhang,Y.,Ding,Y.,Kuai,B,Huang,X.,Mapping and validation of the quantitative trait loci for leaf stay-green-associated parameters in maize.Plant Breeding,2017,136,188-196；Zheng,H.J.,Wu,A.Z.,Zheng,C.C.,Wang,Y.F.,Cai,R.,Shen,X.F.,Xu, r.r., liu, p., kong, l.j., dong, s.t., QTL mapping of size (Zea mays) stand-green traits and their relationship to yield.plant Breeding,2009,128,54-62). The molecular marker is very important for researching the aging characteristic of the corn leaves at the molecular level.
Disclosure of Invention
The first object of the invention is to provide two molecular markers linked with the corn leaf senescence main effect QTL locus.
Another object of the invention is to provide the use of molecular markers linked to the major QTL for maize leaf senescence.
A primer pair for amplifying a molecular marker linked to a major site Stg3 and Stg7 of maize leaf senescence, wherein:
the forward primer 1 sequence of M36 is SEQ ID NO.1, the forward primer 2 sequence is SEQ ID NO.2, and the reverse primer sequence is SEQ ID NO.3;
the forward primer 1 sequence of A-3-11 is SEQ ID NO.4, the forward primer 2 sequence is SEQ ID NO.5, and the reverse primer sequence is SEQ ID NO.6;
the forward primer sequence of A-7-6 is SEQ ID NO.7, and the reverse primer sequence is SEQ ID NO.8;
the forward primer sequence of A-7-7 is SEQ ID NO.9, and the reverse primer sequence is SEQ ID NO.10.
A molecular marker linked to the major site Stg3 and Stg7 of maize leaf senescence, characterized in that the major Stg3 site is located on chromosome 3 of maize and the molecular markers linked to the major Stg3 site are M36 and a-3-11; the main effect Stg7 locus is positioned on a corn chromosome 7, and molecular markers linked with the main effect Stg7 locus are A-7-6 and A-7-7; the specific primer sequences for the amplification of each molecular marker are as follows:
the sequence of the forward primer 1 of M36 is SEQ ID NO.1, the sequence of the forward primer 2 is SEQ ID NO.2, the sequence of the reverse primer is SEQ ID NO.3, and the base of the molecular marker amplification product in the stay-green strain is G;
the sequence of the forward primer 1 of the A-3-11 is SEQ ID NO.4, the sequence of the forward primer 2 is SEQ ID NO.5, the sequence of the reverse primer is SEQ ID NO.6, and the base of the molecular marker amplification product in the green-keeping strain is A;
the forward primer sequence of A-7-6 is SEQ ID NO.7, the reverse primer sequence is SEQ ID NO.8, and the molecular marker amplification product in the stay-green strain is a DNA fragment of 145 bp;
the forward primer sequence of A-7-7 is SEQ ID NO.9, the reverse primer sequence is SEQ ID NO.10, and the molecular marker amplified product is a 144bp DNA fragment in the stay green strain.
The stay-green strain is a near isogenic line NIL-Stg3 constructed by the stay-green strain Si-144 and the aging strain Si-287 Si-144 、NIL-Stg7 Si-144 And NIL-Stg3 Si-144 /Stg7 Si-144 。
The invention is realized by the following technical scheme:
two molecular markers linked with major QTL (Stg 3 and Stg 7) of maize leaf senescence, wherein the major sites Stg3 and Stg7 are respectively positioned on chromosome 3 and chromosome 7 of maize, and F is constructed on early senescence strain Si-287 and green-keeping strain Si-144 of leaves 2 In the population, the contribution rates of Stg3 and Stg7 to the senescent phenotype were 7.40% and 8.41%, respectively. The molecular markers linked to the major sites Stg3 and Stg7 are M36, A-3-11 (Stg 3) and A-7-6, A-7-7 (Stg 7), respectively, and the primer sequences of the molecular markers and the target band lengths or bases amplified in the stay-green strain Si-144 are as follows:
the sequence of the forward primer 1 of M36 is SEQ ID NO.1, the sequence of the forward primer 2 is SEQ ID NO.2, the sequence of the reverse primer is SEQ ID NO.3, and the base detectable in the stay green strain Si-144 is G;
the forward primer 1 of A-3-11 has the sequence of SEQ ID NO.4, the forward primer 2 has the sequence of SEQ ID NO.5, the reverse primer has the sequence of SEQ ID NO.6, and the base detectable in the stay green strain Si-144 is A;
the forward primer sequence of A-7-6 is SEQ ID NO.7, the reverse primer sequence is SEQ ID NO.8, and the DNA fragment with the band size of 145bp can be amplified in the stay green strain Si-144;
a-7-7 has the forward primer sequence of SEQ ID NO.9 and the reverse primer sequence of SEQ ID NO.10, and DNA fragments with the band size of 144bp can be amplified in the stay green strain Si-144.
The application of a molecular marker linked with the main effect sites Stg3 and Stg7 of maize leaf senescence comprises the following steps:
(1) The application in positioning the main effect sites Stg3 and Stg7 of maize leaf senescence;
(2) The application in identifying the senescence character of corn leaves;
(3) The application in screening and identifying the corn senescence-delaying strain.
Extracting corn genome DNA by the application, and amplifying by adopting a primer pair M36, wherein the forward primer 1 sequence is SEQ ID NO.1, the forward primer 2 sequence is SEQ ID NO.2, and the reverse primer sequence is SEQ ID NO.3; or alternatively
The forward primer 1 sequence of A-3-11 is SEQ ID NO.4, the forward primer 2 sequence is SEQ ID NO.5, and the reverse primer sequence is SEQ ID NO.6; or alternatively
The forward primer sequence of A-7-6 is SEQ ID NO.7, and the reverse primer sequence is SEQ ID NO.8; or alternatively
The forward primer sequence of A-7-7 is SEQ ID NO.9, and the reverse primer sequence is SEQ ID NO.10;
wherein: the sequence of the forward primer 1 of M36 is SEQ ID NO.1, the sequence of the forward primer 2 is SEQ ID NO.2, the sequence of the reverse primer is SEQ ID NO.3, and the base of the molecular marker amplification product in the stay-green strain is G; indicating that the main effect site Stg3 of the maize leaf senescence Si-144 The existence of alleles, the strain has the corn leaf senescence delay character, and is a corn leaf senescence delay strain;
the sequence of the forward primer 1 of the A-3-11 is SEQ ID NO.4, the sequence of the forward primer 2 is SEQ ID NO.5, the sequence of the reverse primer is SEQ ID NO.6, and the base of the molecular marker amplification product in the green-keeping strain is A; indicating that the main effect site Stg3 of the maize leaf senescence Si-144 The existence of alleles, the strain has the corn leaf senescence delay character, and is a corn leaf senescence delay strain;
a-7-6 has the forward primer sequence of SEQ ID NO.7, the reverse primer sequence of SEQ ID NO.8, and the molecular marker amplification product in the stay-green strain is 145bpA DNA fragment; indicating that the main effect site Stg7 of the maize leaf senescence Si-144 The existence of alleles, the strain has the corn leaf senescence delay character, and is a corn leaf senescence delay strain;
the forward primer sequence of A-7-7 is SEQ ID NO.9, the reverse primer sequence is SEQ ID NO.10, and the molecular marker amplified product in the stay-green strain is a DNA fragment of 144 bp; indicating that the main effect site Stg7 of the maize leaf senescence Si-144 The existence of the allele, the strain has the corn leaf senescence delay property, and the strain is a corn leaf senescence delay strain.
Specifically, the molecular marker provided by the invention is obtained by the following method:
(1) Constructing a hybridization population by taking a maize premature senility variety Si-287 as a female parent and a green-keeping variety Si-144 as a male parent to obtain a hybridization F 1 And (3) replacing. F (F) 1 Further selfing to obtain F consisting of 207 individual plants 2 At 30DAP (Days After Pollination), leaf yellowing areas of individual plants were investigated and counted, and genotypes of individual plants were detected using 101 KASP markers with polymorphisms between parents. The obtained genotype and phenotype data were imported into QTL icomapping software for QTL analysis, two major sites were obtained, which were located on chromosome 3 (Stg 3) and chromosome 7 (Stg 7) of maize, respectively, with interpretation rates (PVE) of 7.40% and 8.41% for the phenotype, respectively, and LOD values of 3.40 and 4.04, respectively.
(2) At F 2 Selecting a green-retaining single plant as a male parent, continuously hybridizing for three generations by taking Si-287 as a recurrent parent, and selfing for 1 generation to obtain BC 3 F 2 A population. The population was planted in beijing in summer in 2015 and leaf senescence phenotypes were investigated, 30 extreme green-keeping individuals and 30 extreme senescence individuals were selected according to the phenotypes, and genomic DNA was extracted for QTL-seq analysis. The results show that there is one major QTL on chromosome 3 and on chromosome 7, respectively, which are located in the 145.66-161.01Mb and 167.12-170.68Mb intervals on the corresponding chromosomes, respectively, which further verifies the two sites Stg3 and Stg7 obtained by QTL mapping.
(3) To further narrow down the candidate region, we then planted BC 3 F 3 And BC (binary code) 3 F 4 The same 30DAP method is used for investigation and statistics of leaf senescence phenotype of the population, 30 newly developed molecular markers with polymorphism between parents are used for fine localization of target segments, the finally determined candidate regions are 5.86Mb (Stg 3) and 380Kb (Stg 7), and the molecular markers linked with leaf senescence major QTL are determined to be M36, A-3-11 (Stg 3) and A-7-6, A-7-7 (Stg 7).
The beneficial effects of the invention are as follows:
(1) The two leaf senescence QTLs (Stg 3 and Stg 7) positioned by the method are respectively positioned on the No.3 chromosome and the No.7 chromosome of corn, and the physical distance between the two leaf senescence QTLs and the molecular markers linked with the two leaf senescence QTLs is 5.86Mb and 380Kb respectively. The method lays a foundation for further positioning leaf senescence-associated genes and using the leaf senescence-associated genes to carry out molecular marker-assisted selection of senescence-delayed corn varieties in the future.
(2) The invention utilizes F 2 The population (Si-287×Si-144) analyzes the corn leaf senescence-associated QTL and finds two major QTL's, stg3 and Stg7, respectively, the major site Stg3 being able to interpret a phenotypic variation of 7.40% and the major site Stg7 being able to interpret a phenotypic variation of 8.41%.
(3) The molecular marker has important application value in screening or identifying senescence-delaying maize strains, cloning or further finely positioning senescence-associated genes contained in QTL. The invention can be applied to molecular marker assisted breeding, can carry out early generation screening on corn germplasm, overcomes the environmental influence and improves the breeding and selection efficiency.
Drawings
FIG. 1 is the localization of two major QTLs for leaf senescence; wherein,
panel A is a schematic representation of the population construction process for identification of leaf senescence candidate sites. Mainly taking Si-287 as recurrent parent to construct backcross population;
b is F 2 A histogram of the senescent phenotype of the leaves of the population;
panel C is the identification of the leaf senescence regulatory site Stg 3. PVEs stand for F 2 Interpretation rate of candidate sites on phenotypic variation in populations by traditional QTL localizationThe Delta SNP index map of the QTL-seq further localizes Stg3 to the 145.66-161.01Mb region on chromosome 3 (Chr 3), and the candidate region is localized between markers M36 and A-3-11 by linkage analysis of the localization population using molecular markers;
panel D is the identification of the leaf senescence regulatory site Stg7. Delta SNP index map of QTL-seq Stg7 was further determined at the region of 167.12-170.68Mb on chromosome 7 (Chr 7), and the mapped population was subjected to linkage analysis using molecular markers to map this site to the region between A-7-6 and A-7-7.
FIG. 2 field performance of an improved line of Jidan27 obtained by the replacement lines of Stg3 and Stg 7; wherein,
graph A shows the proportion of reduced leaf greenness of each plant line under drought treatment compared with normal watering conditions;
panel B shows the spike weight reduction ratio for each line under drought conditions as compared to normal watering conditions.
Detailed Description
Embodiments of the present invention will be described in detail below with reference to the accompanying drawings, and the embodiments are implemented on the premise of taking the present invention as a technology, and detailed implementation processes and methods are given. The scope of protection of the invention is not limited to this embodiment. The scope of the invention is defined by the appended claims.
The biomaterials used in the examples described below are all commercially available.
Example 1 determination method of molecular markers closely linked to major QTL for maize leaf senescence
Hybridizing aged corn Si-287 as female parent and green corn Si-144 as male parent in F 2 The generation selects green-holding single plant as male parent, si-287 as female parent to be continuously backcrossed for three generations to generate BC 3 F 1 Obtaining BC by selfing the first generation 3 F 2 Self-mating to produce BC after phenotypic and genotypic identification 3 F 3 And BC (binary code) 3 F 4 Population (figure 1).
To record and calculate leaf senescence phenotypes, we divided maize leaf senescence degree (YDL) into five classes of 0, 25%, 50%, 75% and 100%. The number of Leaves (LN) was recorded, and 3 or more individuals per family. Then, the green holding degree (RGL) of the leaf was calculated as rgl=1- (Total YDL)/LN.
F 2 QTL localization of the generation was performed using QTL IciMapping software, using the complete interval mapping method (Inclusive Composite Interval Mapping, ICIM), and the LOD threshold for each site was determined by 1000 permutation tests at a significance level of p=0.05. Analysis found that there was a candidate site on chromosomes 3 and 7 with LOD values of 3.40 and 4.04, respectively, and explained genetic variations of 7.40% and 8.41%, respectively (FIG. 1).
To further verify these two candidate sites, at BC 3 F 2 Instead, we selected 30 individuals with extreme phenotypes (senescence and stay green), extracted genomic DNA by CTAB method and subjected to mixed pool sequencing (senescence DNA pool C1 and stay green DNA pool C2), high quality reads obtained by sequencing were aligned to a reference genome, SNP variation data were extracted and SNP-index was calculated, sites with sequencing depth less than 7 and SNP-index less than 0.3 were removed, 4817475 variation sites were finally obtained, and delta (SNP-index) of all sites was calculated by the following formula: delta (SNP-index) =snp-index (C2) -SNP-index (C1). At a 99% level of significance, major QTLs were detected on chromosome 3 (145.66-161.01 Mb) and chromosome 7 (167.12-170.68 Mb), respectively, which overlapped the major regions of Stg3 and Stg7, the QTL mapping derived sites, indicating that these two sites have important roles in leaf senescence regulation. To further narrow the target segment we utilized the newly developed 30 molecular marker pairs BC 3 F 3 And BC (binary code) 3 F 4 The population was examined, and Stg3 was finally located between molecular markers M36 and A-3-11, and Stg7 was located between molecular markers A-7-6 and A-7-7. The two pairs of molecular markers closely linked with the candidate sites lay a foundation for screening senescence-delaying strains.
In Stg3, the forward primer sequences 1 and 2 of KASP/M36 are SEQ ID NO.1 and SEQ ID NO.2 respectively, the reverse primer sequence is SEQ ID NO.3, and in green-keeping strains Si-144 and NIL-Stg3 Si-144 And NIL-Stg3 Si-144 /Stg7 Si-144 In (3) can be amplified to obtainThe base of (2) is G; the forward primer sequence 1 and the forward primer sequence 2 of KASP/A-3-11 are SEQ ID NO.4 and SEQ ID NO.5 respectively, the reverse primer sequence is SEQ ID NO.6, and the forward primer sequence and the reverse primer sequence are respectively used for preparing the sequence of the KASP/A-3-11 in green-keeping strains Si-144 and NIL-Stg3 Si-144 And NIL-Stg3 Si-144 /Stg7 Si-144 The amplified base is A (Table 1); in Stg7, inDel/A-7-6 has a forward primer sequence of SEQ ID NO.7 and a reverse primer sequence of SEQ ID NO.8, and is used in green-keeping strains Si-144 and NIL-Stg7 Si-144 And NIL-Stg3 Si-144 /Stg7 Si-144 In the method, a DNA fragment with a band size of 145bp can be amplified; the forward primer sequence of InDel/A-7-7 is SEQ ID NO.9, the reverse primer sequence is SEQ ID NO.10, and the InDel/A-7-7 is a green-keeping strain Si-144 and NIL-Stg7 Si-144 And NIL-Stg3 Si-144 /Stg7 Si-144 In (2), a DNA fragment with a band size of 144bp was amplified (Table 2).
TABLE 1 genotyping of molecular markers of the invention closely linked to Stg3 in different strains
TABLE 2 genotyping of molecular markers of the invention closely linked to Stg7 in different lines
Example 2 modification of Gemini 27 with a substitution line carrying QTL fragments Stg3 and Stg7
Using the two pairs of molecular markers M36, A-3-11 and A-7-6, A-7-7 pairs BC in the present invention 3 F 4 Genotyping the individual plants in the population to obtain 6 individual plants with a phenotype and a genotype of stay green type (aging delay), including two NIL-Stg3 Si-144 Two NIL-Stg7 Si-144 And two NIL-Stg3 Si-144 /Stg7 Si-144 . The background of these lines was examined using 104 KASP markers and showed that their background recovery (Si-287 background) was 90.38% -98.1%. The 6 strains are respectively taken as female parent, andhybridization is carried out by taking Si-144 as male parent to obtain improved hybrid F 1 Generation (modified line of Jidan 27).
The hybrid seeds and Jidan27 (hybrid F obtained by hybridization with Si-287 as female parent and Si-144 as male parent) are planted in Jinta county (39 DEG 57'N,98 DEG 22' E) and inner Mongolian Bayan (40 DEG 75'N,107 DEG 42' E) in Gansu province in 2017 1 Generation), these two sites are drought and rain-free during the growing season of the plants. Setting two test areas of normal watering and drought treatment, wherein the normal watering area is managed according to a local field management mode, the drought treatment area stops watering from one week before flowering to 45 days after flowering, and other management modes are consistent with the normal watering area. Investigation of leaf senescence degree in the field was conducted 45 days after flowering, and NIL-Stg3 was compared with the normally watered area Si-144 ×Si-144、NIL-Stg7 Si-144 X Si-144 and NIL-Stg3 Si-144 /Stg7 Si-144 The blade retention green reduction ratio of x Si-144 was lower than that of jican 27 (fig. 2A), and the spike weight (EW) reduction ratio also showed a similar trend (fig. 2B). This shows that the two QTLs Stg3 and Stg7 not only play an important role in improving the green retention of the leaves, but also contribute to stable yield under drought conditions.
The foregoing description is only illustrative of the present invention and is not intended to limit the scope of the invention, and all equivalent structures or equivalent processes or direct or indirect application in other related arts are included in the scope of the present invention.
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Claims (4)
1. The application of a molecular marker linked with a main effect site Stg7 of maize leaf senescence in positioning the main effect site Stg7 of maize leaf senescence, wherein the main effect site Stg7 is positioned on a chromosome 7 of maize, and the molecular marker linked with the main effect site Stg7 is A-7-6 or A-7-7; the specific primer sequences for the amplification of each molecular marker are as follows:
the forward primer sequence of A-7-6 is SEQ ID NO.7, and the reverse primer sequence is SEQ ID NO.8;
the forward primer sequence of A-7-7 is SEQ ID NO.9, and the reverse primer sequence is SEQ ID NO.10;
the corn is near isogenic line NIL-Stg7 constructed by stay-green strain Si-144 and stay-green strain Si-287 Si-144 。
2. Application of a molecular marker linked with a main effect site Stg7 of maize leaf senescence in identifying maize leaf senescence traits;
the main effect site Stg7 is positioned on a corn chromosome 7, and a molecular marker linked with the main effect site Stg7 is A-7-6 or A-7-7; the specific primer sequences for the amplification of each molecular marker are as follows:
the forward primer sequence of A-7-6 is SEQ ID NO.7, and the reverse primer sequence is SEQ ID NO.8;
the forward primer sequence of A-7-7 is SEQ ID NO.9, and the reverse primer sequence is SEQ ID NO.10;
the corn is near isogenic line NIL-Stg7 constructed by stay-green strain Si-144 and stay-green strain Si-287 Si-144 。
3. Application of a molecular marker linked with a main effect site Stg7 of maize leaf senescence in screening and identifying maize leaf senescence delay lines;
the main effect site Stg7 is positioned on a corn chromosome 7, and a molecular marker linked with the main effect site Stg7 is A-7-6 or A-7-7; the specific primer sequences for the amplification of each molecular marker are as follows:
the forward primer sequence of A-7-6 is SEQ ID NO.7, the reverse primer sequence is SEQ ID NO.8,
the forward primer sequence of A-7-7 is SEQ ID NO.9, and the reverse primer sequence is SEQ ID NO.10;
the corn is near isogenic line NIL-Stg7 constructed by stay-green strain Si-144 and stay-green strain Si-287 Si-144 。
4. A use according to any one of claims 1-3, characterized in that: extracting corn genome DNA, amplifying by using the following primer pair,
the forward primer sequence of A-7-6 is SEQ ID NO.7, and the reverse primer sequence is SEQ ID NO.8; or alternatively
The forward primer sequence of A-7-7 is SEQ ID NO.9, and the reverse primer sequence is SEQ ID NO.10;
the forward primer sequence of A-7-6 is SEQ ID NO.7, the reverse primer sequence is SEQ ID NO.8, and the molecular marker amplification product in the stay-green strain is a DNA fragment of 145 bp; indicating that the main effect site Stg7 of the maize leaf senescence Si-144 The existence of alleles, the strain has the corn leaf senescence delay character, and is a corn leaf senescence delay strain;
the forward primer sequence of A-7-7 is SEQ ID NO.9, the reverse primer sequence is SEQ ID NO.10, and the molecular marker amplified product in the stay-green strain is a DNA fragment of 144 bp; indicating that the main effect site Stg7 of the maize leaf senescence Si-144 The existence of the allele, the strain has the corn leaf senescence delay property, and the strain is a corn leaf senescence delay strain.
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| CN101519692A (en) * | 2009-02-25 | 2009-09-02 | 上海市农业科学院 | Molecular mark closely interlocked with green-keeping main effect QTL of corn leaf blade |
| WO2016139657A1 (en) * | 2015-03-01 | 2016-09-09 | Y. E. Vigor. Corn | Markers for high yield in maize |
| CN109295248A (en) * | 2018-10-26 | 2019-02-01 | 河南省农业科学院作物设计中心 | For detecting primer, kit, detection method and the application of the molecular labeling of control corn stem intensity main effect QTL linkage |
| CN109836482A (en) * | 2017-11-28 | 2019-06-04 | 中国农业大学 | Corn gene KRN2 and application thereof |
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| CN101519692A (en) * | 2009-02-25 | 2009-09-02 | 上海市农业科学院 | Molecular mark closely interlocked with green-keeping main effect QTL of corn leaf blade |
| WO2016139657A1 (en) * | 2015-03-01 | 2016-09-09 | Y. E. Vigor. Corn | Markers for high yield in maize |
| CN109836482A (en) * | 2017-11-28 | 2019-06-04 | 中国农业大学 | Corn gene KRN2 and application thereof |
| CN109295248A (en) * | 2018-10-26 | 2019-02-01 | 河南省农业科学院作物设计中心 | For detecting primer, kit, detection method and the application of the molecular labeling of control corn stem intensity main effect QTL linkage |
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