CN104694625B - A kind of method of utilization molecular marking supplementary breeding length grain type paddy rice - Google Patents
A kind of method of utilization molecular marking supplementary breeding length grain type paddy rice Download PDFInfo
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Abstract
The invention discloses a kind of method of utilization molecular marking supplementary breeding length grain type paddy rice：Rice varieties Lemont is built into segregating populations with No. 4 hybridization of rice are raised, 4 QTLs related to grain length are detected respectively using 4 molecular labelings such as D309, RM3585, D463, D755 to the individual plant in segregating population：qGL‑3‑1YD、qGL‑3‑2LE、qGL‑4YD、qGL‑7LE.D309 No. 4 molecular labeling banding patterns of rice of raising amplified are named as A, the Lemont banding patterns of RM3585 amplifications are named as B, and No. 4 banding patterns of rice of raising of D463 amplifications are named as C, and the Lemont banding patterns of D755 amplifications are named as D.The individual plant of different number of genotype in tetra- molecular marker gene types of selective polymerization A, B, C, D is needed to obtain the rice breed of different grain lengths according to breeding.Patent of the present invention can not by phenotypic evaluation seed selection difference grain length long grain type rice material, to improve breeding efficiency.
Description
Technical field
The present invention relates to it is a kind of using molecular marker assisted selection come selective polymerization it is different make paddy rice grain length increase number
Measure character gene seat (QTL), in the method for the long grain type paddy rice of the different grain lengths of seed selection.
Background technology
Paddy rice is the important cereal crops of China, and Rice Production is related to national economy and national food security.In China,
With steadily improving for rice yield, rice breeding work it is also proposed higher and higher requirement to paddy rice exterior quality.Grain
Length (abbreviation grain length) is an important economical character of relation paddy rice exterior quality.The japonica rice variety of northern China plantation is more
For short round kind, northern resident is also inclined to consume short grain type rice；The long-grained nonglutinous rice planted in south China is generally slender type kind,
Southern resident (the particularly province such as Guangdong, Guangxi, Fujian) the long grain type rice of deviation consumption more.The different grain length water of traditional seed selection
The method of rice be by phenotypic evaluation carry out, it is necessary to be selected after paddy rice is ripe grain length.By molecular labeling to water
The genotype of rice, which carries out selection, to be selected in rice seedling grain length, not only improve the accuracy of selection, Er Qieti
High breeding efficiency.Paddy rice grain length is a quantitative character, is controlled by quantitative trait locus position (QTL).At present, although paddy rice
The substantial amounts of QTL related to rice grain length has been oriented in scientific research circle, also obtains many molecular labelings with its close linkage, but
It is how these QTLs related to grain length being positioned on paddy rice coloured differently body combine and use, using what molecule mark
Different grain length QTL is effectively polymerize by noteAnswer to these problems, can effectively instruct breeding practice, improve
Breeding efficiency simultaneously accelerates breeding process.In this context, the present invention propose one kind polymerize 4 using molecular marker assisted selection
The individual QTL for increasing paddy rice grain length, in the method for seed selection length grain type paddy rice, the present invention has more compared with traditional Phenotypic Selection method
High selection accuracy, and breeding efficiency can be improved.
The content of the invention
The present invention use using molecular marker assisted selection come selectively polymerize it is multiple make paddy rice grain length increase
QTL, in the method for seed selection length grain type paddy rice.Which, which polymerize, present invention mainly solves (a) makes the increased QTL of grain length(b)
Which selected using molecular labelingThe two problems.The invention provides one kind polymerization qGL-3-1YD, qGL-3-2LE,
QGL-4YD, qGL-7LE etc. 4 makes the increased QTL of paddy rice grain length, and utilizes 4 molecules such as D309, RM3585, D463, D755
Mark the method that selection is carried out to this 4 QTL respectively.The present invention effectively make use of qGL-3-1YD, qGL-3-2LE, qGL-
4YD, qGL-7LE etc. 4 makes the QTL of grain length growth additive effect, and effectively make use of D309, RM3585, D463, D755
Deng effectiveness of selection of 4 molecular labelings to this 4 QTL, the different grain length QTL of accurate polymerization is reached, with seed selection length grain type
The purpose of paddy rice.
The present invention comprises the following steps：
(1) rice varieties Lemont and rice varieties are raised into No. 4 hybridization of rice and builds segregating population Fn (n >=2)；
(2) segregating population that PCR amplification steps (1) are obtained is distinguished with 4 molecular labelings D309, RM3585, D463, D755
The leaf DNA of interior individual plant；
(3) using the segregating population of 4 molecular labelings difference detecting step (1) description described in step (2), 4 are followed the trail of
The quantitative trait locus position (QTL) related to rice grain length；Wherein mark D309 is positioned at the chromosome of paddy rice the 3rd, for selecting
Select the QTL relevant with grain length growth positioned at the chromosome of paddy rice the 3rd：qGL-3-1YD；RM3585 is positioned at the chromosome of paddy rice the 3rd,
For selecting the QTL relevant with grain length growth positioned at the chromosome of paddy rice the 3rd：qGL-3-2LE；D463 is positioned at the dye of paddy rice the 4th
Colour solid, for selecting the QTL relevant with grain length growth positioned at the chromosome of paddy rice the 4th：qGL-4YD；D755 is positioned at paddy rice the 7th
Chromosome, for selecting the QTL relevant with grain length growth positioned at the chromosome of paddy rice the 7th：qGL-7LE；D309 is expanded
A is named as with raising No. 4 banding pattern identical molecular marker gene types of rice, what RM3585 was expanded divides with Lemont banding patterns identical
Sub- marker genetype is named as B, and what D463 was expanded is named as C with raising No. 4 banding pattern identical molecular marker gene types of rice, will
What D755 was expanded is named as D with Lemont banding pattern identical molecular marker gene types；
(4) whole four genotype of A, B, C, D that polymerization procedure (3) is described in the segregating population of selection step (1) description
Individual plant to obtain the rice material of maximum grain length；Select step (1) description segregating population in polymerization procedure (3) description A,
The individual plant of any 3 genotype in tetra- genotype of B, C, D is to obtain the rice material that grain length takes second place；Selection step (1) is retouched
The individual plant of any 2 genotype in tetra- genotype of A, B, C, D that polymerization procedure (3) is described in the segregating population stated is to obtain
The grain length rice material shorter compared with the individual plant for polymerizeing any 3 genotype；Select to carry step in the segregating population of step (1) description
Suddenly the individual plant of any 1 genotype in tetra- genotype of A, B, C, D of (3) description relatively polymerize any 2 bases to obtain grain length
Because of the shorter rice material of the individual plant of type；
(5) the individual plant selfing many generations for the different number of genotype for describing polymerization procedure (4), to obtain different grain lengths
Long grain type paddy rice homozygosis material.
Further, in the step (1), Lemont is hybridized with raising rice No. 4, Lemont may be selected and is used as female parent
Rice No. 4 is raised as male parent, can also select to raise rice No. 4 and hybridized as maternal Lemont as male parent.
Further, in the step (2), segregating population is detected with 4 molecular labelings D309, RM3585, D463, D755
The number of interior individual plant should be determined according to actual needs；By taking F2 colonies as an example, want step (4) screening obtain 1 plant carry A,
B, C, D all four genotype paddy rice individual plant, should detect in theory no less than 256 individual plants (computational methods be 4 × 4 × 4 ×
4)；For example, want the paddy rice individual plant of 3 genotype in step (4) screens 1 plant of carrying tetra- genotype of A, B, C, D of acquisition,
No less than 64 individual plants should be detected in theory (computational methods are 4 × 4 × 4)；Certainly, this is theoretic supposition, in reality
In detection process, it should determine the individual plant number of detection as needed, generally speaking, the individual plant number of detection is more, then obtains
The chance of the different desired individual plants of step (4) description is just bigger.
Further, in the step (3), molecular marker gene type A is the molecule for raising rice No. 4 detected using D309
Marker bands, but A is not qGL-3-1YD genotype, can be indirectly by choosing due to D309 and qGL-3-1YD gene linkages
A genotype is selected to select qGL-3-1YD genotype；It can similarly be passed through according to RM3585 and qGL-3-2LE gene linkages
Indirect selections 1 B gene type selects qGL-3-2LE genotype；A selecting is passed through according to D463 and qGL-4YD gene linkages
C genotype is selected to select qGL-4YD genotype；Indirect selections D genotype is passed through according to D755 and qGL-7LE gene linkages
To select qGL-7LE genotype.
Further, in the step (4), it polymerize the average grain length of individual plant of different number of genotype by from big to small
It is ordered as：Any 2 ＞ are comprised only in any 3 ＞ polymerizations A, B, C, D in whole 4 ＞ polymerizations A, B, C, the D of polymerization A, B, C, D
Any 1 in A, B, C, D；It is pointed out that above-mentioned sequence size is only applicable to be planted under same environmental condition to same
Segregating population in the one Fn generations (n is fixed numbers) is compared.
Further, in the step (5), selfing algebraically should be actually needed determination according to breeding.
Further, the non-groat length that the long grain type paddy rice that patent of the present invention is mentioned is defined as not including awns is more than
9mm or so paddy rice.
Embodiment
Embodiment 1：Selection carries the individual plant of any one genotype in tetra- genotype of A, B, C, D to obtain average grain
The long F2 more than 9.45mm is for rice material
1. rice varieties Lemont is made into maternal, raise rice No. 4 with rice varieties and make male parent and hybridized and build F2 to separate
(rice varieties are raised rice No. 4 and carried with rice varieties Lemont by the national Rice Germplasm Resources mid-term storehouse of China Paddy Rice Inst for colony
For).
2. distinguish 191 lists in PCR amplification F2 segregating populations with 4 molecular labelings D309, RM3585, D463, D755
Strain, this F2 colony including 191 individual plants including sows in May, 2011 and tried in China Paddy Rice Inst of Hangzhou Fuyang City
Test farm；D309, D463, D755 flag sequence are shown in Zeng Y.X., et al.Development of
1047insertion-deletion markers for rice genetic studies and breeding.Genetics
And Molecular Research, 2013, volume 12：5226-5235；RM3585 flag sequence is from http：//
Www.gramene.org/ public databases are obtained.
3. qGL-3-1YD genes (the molecule mark of A genotype for raising rice No. 4 is followed the trail of by D309 marks tracking A genotype
Note banding pattern is that D309 marks the banding pattern for expanding and raising rice No. 4)；Follow the trail of 1 B gene type to follow the trail of Lemont's by RM3585 marks
QGL-3-2LE genes (the molecular labeling banding pattern of 1 B gene type is the banding pattern that RM3585 marks amplification Lemont)；Marked by D463
Follow the trail of C genotype come follow the trail of raise rice No. 4 qGL-4YD genes (the molecular labeling banding pattern of C genotype be D463 mark amplification raise rice
The banding pattern of No. 4)；Follow the trail of D genotype to follow the trail of the Lemont qGL-7LE genes (molecular labeling of D genotype by D755 marks
Banding pattern is the banding pattern that D755 marks amplification Lemont).
4. the individual plant that selection carries any 1 genotype in tetra- genotype of A, B, C, D can obtain grain length in F2 generations
More than the rice material (table 1) of 9.45mm (average value).
Table 1 selects the individual plant of obtained carrying different genotype in F2 using 4 different molecular labelings progress from generation to generation
Grain length average (n represents to carry the individual plant number of corresponding gene type)
| Genotype (mark used) | Grain length (mm) |
| Carry A (D309) | 9.45 ± 0.45 (n=37) |
| Carry B (RM3585) | 9.48 ± 0.43 (n=41) |
| Carry C (D463) | 9.47 ± 0.41 (n=59) |
| Carry D (D755) | 9.45 ± 0.45 (n=40) |
5. generation more than the paddy rice individual plant selfing that will be selected in, to obtain the rice strain that other economical characters are stable.
Embodiment 2：The individual plant of any 2 genotype in tetra- genotype of selective polymerization A, B, C, D is to obtain average grain
The long F2 more than 9.61mm is for rice material
1. rice varieties Lemont is made into maternal, raise rice No. 4 with rice varieties and make male parent and hybridized and build F2 to separate
(rice varieties are raised rice No. 4 and carried with rice varieties Lemont by the national Rice Germplasm Resources mid-term storehouse of China Paddy Rice Inst for colony
For).
2. distinguish 191 lists in PCR amplification F2 segregating populations with 4 molecular labelings D309, RM3585, D463, D755
Strain, this F2 colony including 191 individual plants including sows in May, 2011 and tried in China Paddy Rice Inst of Hangzhou Fuyang City
Test farm；D309, D463, D755 flag sequence are shown in Zeng Y.X., et al.Development of 1047insertion-
deletion markers for rice genetic studies and breeding.Genetics and Molecular
Research, 12 in 2013：5226-5235；RM3585 flag sequence is from http：The public numbers of //www.gramene.org/
Obtained according to storehouse.
3. qGL-3-1YD genes (the molecule mark of A genotype for raising rice No. 4 is followed the trail of by D309 marks tracking A genotype
Note banding pattern is that D309 marks the banding pattern for expanding and raising rice No. 4)；Follow the trail of 1 B gene type to follow the trail of Lemont's by RM3585 marks
QGL-3-2LE genes (the molecular labeling banding pattern of 1 B gene type is the banding pattern that RM3585 marks amplification Lemont)；Marked by D463
Follow the trail of C genotype come follow the trail of raise rice No. 4 qGL-4YD genes (the molecular labeling banding pattern of C genotype be D463 mark amplification raise rice
The banding pattern of No. 4)；Follow the trail of D genotype to follow the trail of the Lemont qGL-7LE genes (molecular labeling of D genotype by D755 marks
Banding pattern is the banding pattern that D755 marks amplification Lemont).
4. the individual plant of any 2 genotype in tetra- genotype of selective polymerization A, B, C, D can obtain grain length and be more than
9.61mm (average value) rice material (table 2).
Table 2 utilizes any 2 among 4 molecular labeling difference selective polymerization tetra- genotype of A, B, C, D from generation to generation in F2
The grain length average of the individual plant of genotype (n represents to carry the individual plant number of corresponding gene type)
| D309 | RM3585 | D463 | D755 | Grain length (mm) |
| Carry A | Carry B | - | - | 9.67 ± 0.42 (n=10) |
| Carry A | - | Carry C | - | 9.61 ± 0.34 (n=10) |
| Carry A | - | - | Carry D | 9.64 ± 0.57 (n=9) |
| - | Carry B | Carry C | - | 9.65 ± 0.42 (n=12) |
| - | Carry B | - | Carry D | 9.67 ± 0.34 (n=12) |
| - | - | Carry C | Carry D | 9.74 ± 0.40 (n=15) |
5. generation more than the paddy rice individual plant selfing that will be selected in, to obtain the rice strain that other economical characters are stable.
Embodiment 3：The individual plant of any 3 genotype in tetra- genotype of selective polymerization A, B, C, D is to obtain average grain
The long F2 more than 9.8mm is for rice material
1. rice varieties Lemont is made into maternal, raise rice No. 4 with rice varieties and make male parent and hybridized and build F2 to separate
(rice varieties are raised rice No. 4 and carried with rice varieties Lemont by the national Rice Germplasm Resources mid-term storehouse of China Paddy Rice Inst for colony
For).
2. distinguish 191 lists in PCR amplification F2 segregating populations with 4 molecular labelings D309, RM3585, D463, D755
Strain, this F2 colony including 191 individual plants including sows in May, 2011 and tried in China Paddy Rice Inst of Hangzhou Fuyang City
Test farm；D309, D463, D755 flag sequence are shown in Zeng Y.X., et a1.Development of 1047insertion-
deletion markers for rice genetic studies and breeding.Genetics and Molecular
Research, 12 in 2013：5226-5235；RM3585 flag sequence is from http：The public numbers of //www.gramene.org/
Obtained according to storehouse.
3. qGL-3-1YD genes (the molecule mark of A genotype for raising rice No. 4 is followed the trail of by D309 marks tracking A genotype
Note banding pattern is that D309 marks the banding pattern for expanding and raising rice No. 4)；Follow the trail of 1 B gene type to follow the trail of Lemont's by RM3585 marks
QGL-3-2LE genes (the molecular labeling banding pattern of 1 B gene type is the banding pattern that RM3585 marks amplification Lemont)；Marked by D463
Follow the trail of C genotype come follow the trail of raise rice No. 4 qGL-4YD genes (the molecular labeling banding pattern of C genotype be D463 mark amplification raise rice
The banding pattern of No. 4)；Follow the trail of D genotype to follow the trail of the Lemont qGL-7LE genes (molecular labeling of D genotype by D755 marks
Banding pattern is the banding pattern that D755 marks amplification Lemont).
It is big that 4. the individual plant of any 3 genotype in tetra- genotype of selective polymerization A, B, C, D can obtain average grain length
In 9.8mm rice material (table 3).
Table 3 utilizes any 3 among 4 molecular labeling difference selective polymerization tetra- genotype of A, B, C, D from generation to generation in F2
The grain length average of the individual plant of genotype (n represents to carry the individual plant number of corresponding gene type)
| D309 | RM3585 | D463 | D755 | Grain length (mm) |
| Carry A | Carry B | Carry C | - | 9.80 ± 0 (n=2) |
| Carry A | Carry B | - | Carry D | 9.90 ± 0.14 (n=2) |
| Carry A | - | Carry C | Carry D | 10.00 ± 0.17 (n=3) |
| - | Carry B | Carry C | Carry D | 9.85 ± 0.31 (n=6) |
5. generation more than the paddy rice individual plant selfing that will be selected in, to obtain the rice strain that other economical characters are stable.
Embodiment 4：Selection carries the individual plant of any one genotype in tetra- genotype of A, B, C, D to obtain average grain
The long F7 more than 9.64mm is for rice material
1. rice varieties Lemont is made into maternal, raise rice No. 4 with rice varieties and make male parent and hybridized and build F7 to separate
(rice varieties are raised rice No. 4 and carried with rice varieties Lemont by the national Rice Germplasm Resources mid-term storehouse of China Paddy Rice Inst for colony
For).
2. distinguish 222 lists in PCR amplification F7 segregating populations with 4 molecular labelings D309, RM3585, D463, D755
Strain, this F7 colony including 222 individual plants including sows in May, 2014 and tried in China Paddy Rice Inst of Hangzhou Fuyang City
Test farm；D309, D463, D755 flag sequence are shown in Zeng Y.X., et al.Development of 1047insertion-
deletion markers for rice genetic studies and breeding.Genetics and Molecular
Research, 2013, volume 12：5226-5235；RM3585 flag sequence is from http：//www.gramene.org/ data
Storehouse is obtained.
3. qGL-3-1YD genes (the molecule mark of A genotype for raising rice No. 4 is followed the trail of by D309 marks tracking A genotype
Note banding pattern is that D309 marks the banding pattern for expanding and raising rice No. 4)；Follow the trail of 1 B gene type to follow the trail of Lemont's by RM3585 marks
QGL-3-2LE genes (the molecular labeling banding pattern of 1 B gene type is the banding pattern that RM3585 marks amplification Lemont)；Marked by D463
Follow the trail of C genotype come follow the trail of raise rice No. 4 qGL-4YD genes (the molecular labeling banding pattern of C genotype be D463 mark amplification raise rice
The banding pattern of No. 4)；Follow the trail of D genotype to follow the trail of the Lemont qGL-7LE genes (molecular labeling of D genotype by D755 marks
Banding pattern is the banding pattern that D755 marks amplification Lemont).
4. the individual plant that selection carries any 1 genotype in tetra- genotype of A, B, C, D can obtain grain length in F7 generations
More than the rice material (table 4) of 9.64mm (average value).
Any 1 gene of the table 4 in F7 carries tetra- genotype of A, B, C, D using different Marker-assisted selections from generation to generation
The grain length average of the individual plant of type (n represents to carry the individual plant number of corresponding gene type)
| Genotype (mark used) | Grain length (mm) |
| Carry A (D309) | 9.64 ± 0.76 (n=139) |
| Carry B (RM3585) | 9.75 ± 0.77 (n=94) |
| Carry C (D463) | 9.79 ± 0.70 (n=113) |
| Carry D (D755) | 9.72 ± 0.76 (n=77) |
5. generation more than the paddy rice individual plant selfing on demand that will be selected in, to obtain the paddy rice strain that other economical characters are highly stable
System.
Embodiment 5：The individual plant of any 2 genotype in tetra- genotype of selective polymerization A, B, C, D is to obtain average grain
The long F7 more than 9.84mm is for rice material
1. rice varieties Lemont is made into maternal, raise rice No. 4 with rice varieties and make male parent and hybridized and build F7 to separate
(rice varieties are raised rice No. 4 and carried with rice varieties Lemont by the national Rice Germplasm Resources mid-term storehouse of China Paddy Rice Inst for colony
For).
2. distinguish 222 lists in PCR amplification F7 segregating populations with 4 molecular labelings D309, RM3585, D463, D755
Strain, this F7 colony including 222 individual plants including sows in May, 2014 and tried in China Paddy Rice Inst of Hangzhou Fuyang City
Test farm；D309, D463, D755 flag sequence are shown in Zeng Y.X., et al.Development of 1047insertion-
deletion markers for rice genetic studies and breeding.Genetics and Molecular
Research, 2013, volume 12：5226-5235；RM3585 flag sequence is from http：//www.gramene.org/ data
Storehouse is obtained.
3. qGL-3-1YD genes (the molecule mark of A genotype for raising rice No. 4 is followed the trail of by D309 marks tracking A genotype
Note banding pattern is that D309 marks the banding pattern for expanding and raising rice No. 4)；Follow the trail of 1 B gene type to follow the trail of Lemont's by RM3585 marks
QGL-3-2LE genes (the molecular labeling banding pattern of 1 B gene type is the banding pattern that RM3585 marks amplification Lemont)；Marked by D463
Follow the trail of C genotype come follow the trail of raise rice No. 4 qGL-4YD genes (the molecular labeling banding pattern of C genotype be D463 mark amplification raise rice
The banding pattern of No. 4)；Follow the trail of D genotype to follow the trail of the Lemont qGL-7LE genes (molecular labeling of D genotype by D755 marks
Banding pattern is the banding pattern that D755 marks amplification Lemont).
4. the individual plant of any 2 genotype in tetra- genotype of selective polymerization A, B, C, D can obtain grain length in F7 generations
More than the rice material (table 5) of 9.84mm (average value)；
Grain length average (the n of the individual plant of any 2 genotype of the table 5 in F7 from generation to generation polymerization tetra- genotype of A, B, C, D
Represent the individual plant number of carrying corresponding gene type)
| D309 | RM3585 | D463 | D755 | Grain length (mm) |
| Carry A | Carry B | - | - | 9.88 ± 0.80 (n=56) |
| Carry A | - | Carry C | - | 9.90 ± 0.68 (n=71) |
| Carry A | - | - | Carry D | 9.89 ± 0.77 (n=50) |
| - | Carry B | Carry C | - | 10.07 ± 0.81 (n=42) |
| - | Carry B | - | Carry D | 9.84 ± 0.87 (n=40) |
| - | - | Carry C | Carry D | 9.98 ± 0.78 (n=37) |
5. generation more than the paddy rice individual plant selfing on demand that will be selected in, to obtain the paddy rice strain that other economical characters are highly stable
System.
Embodiment 6：The individual plant of any 3 genotype in tetra- genotype of selective polymerization A, B, C, D is to obtain average grain
The long F7 more than 10.04mm is for rice material
1. rice varieties Lemont is made into maternal, raise rice No. 4 with rice varieties and make male parent and hybridized and build F7 to separate
(rice varieties are raised rice No. 4 and carried with rice varieties Lemont by the national Rice Germplasm Resources mid-term storehouse of China Paddy Rice Inst for colony
For).
2. distinguish 222 lists in PCR amplification F7 segregating populations with 4 molecular labelings D309, RM3585, D463, D755
Strain, this F7 colony including 222 individual plants including sows in May, 2014 and tried in China Paddy Rice Inst of Hangzhou Fuyang City
Test farm；D309, D463, D755 flag sequence are shown in Zeng Y.X., et al.Development of 1047insertion-
deletion markers for rice genetic studies and breeding.Genetics and Molecular
Research, 2013, volume 12：5226-5235；RM3585 flag sequence is from http：//www.gramene.org/ data
Storehouse is obtained.
3. qGL-3-1YD genes (the molecule mark of A genotype for raising rice No. 4 is followed the trail of by D309 marks tracking A genotype
Note banding pattern is that D309 marks the banding pattern for expanding and raising rice No. 4)；Follow the trail of 1 B gene type to follow the trail of Lemont's by RM3585 marks
QGL-3-2LE genes (the molecular labeling banding pattern of 1 B gene type is the banding pattern that RM3585 marks amplification Lemont)；Marked by D463
Follow the trail of C genotype come follow the trail of raise rice No. 4 qGL-4YD genes (the molecular labeling banding pattern of C genotype be D463 mark amplification raise rice
The banding pattern of No. 4)；Follow the trail of D genotype to follow the trail of the Lemont qGL-7LE genes (molecular labeling of D genotype by D755 marks
Banding pattern is the banding pattern that D755 marks amplification Lemont).
4. the individual plant of any 3 genotype in tetra- genotype of selective polymerization A, B, C, D can obtain grain length in F7 generations
More than the rice material (table 6) of 10.04mm (average value)；
The grain length of the individual plant of any 3 genotype of the table 6 in F7 generation selective polymerizations tetra- genotype of A, B, C, D is averaged
Number (n represents to carry the individual plant number of corresponding gene type)
| D309 | RM3585 | D463 | D755 | Grain length (mm) |
| Carry A | Carry B | Carry C | - | 10.27 ± 0.77 (n=26) |
| Carry A | Carry B | - | Carry D | 10.20 ± 0.83 (n=22) |
| Carry A | - | Carry C | Carry D | 10.20 ± 0.67 (n=26) |
| - | Carry B | Carry C | Carry D | 10.04. ± 1.05 (n=18) |
5. generation more than the paddy rice individual plant selfing on demand that will be selected in, to obtain the paddy rice strain that other economical characters are highly stable
System.
Embodiment 7：The individual plant of whole four genotype of selective polymerization A, B, C, D is to obtain average grain length as 10.53mm's
F7 is for rice material
1. rice varieties Lemont is made into maternal, raise rice No. 4 with rice varieties and make male parent and hybridized and build F7 to separate
(rice varieties are raised rice No. 4 and carried with rice varieties Lemont by the national Rice Germplasm Resources mid-term storehouse of China Paddy Rice Inst for colony
For).
2. distinguish 222 lists in PCR amplification F7 segregating populations with 4 molecular labelings D309, RM3585, D463, D755
Strain, this F7 colony including 222 individual plants including sows in May, 2014 and tried in China Paddy Rice Inst of Hangzhou Fuyang City
Test farm；D309, D463, D755 flag sequence are shown in Zeng Y.X., et al.Development of 1047insertion-
deletion markers for rice genetic studies and breeding.Genetics and Molecular
Research, 2013, volume 12：5226-5235；RM3585 flag sequence is from http：//www.gramene.org/ data
Storehouse is obtained.
3. qGL-3-1YD genes (the molecule mark of A genotype for raising rice No. 4 is followed the trail of by D309 marks tracking A genotype
Note banding pattern is that D309 marks the banding pattern for expanding and raising rice No. 4)；Follow the trail of 1 B gene type to follow the trail of Lemont's by RM3585 marks
QGL-3-2LE genes (the molecular labeling banding pattern of 1 B gene type is the banding pattern that RM3585 marks amplification Lemont)；Marked by D463
Follow the trail of C genotype come follow the trail of raise rice No. 4 qGL-4YD genes (the molecular labeling banding pattern of C genotype be D463 mark amplification raise rice
The banding pattern of No. 4)；Follow the trail of D genotype to follow the trail of the Lemont qGL-7LE genes (molecular labeling of D genotype by D755 marks
Banding pattern is the banding pattern that D755 marks amplification Lemont).
It is 10.53mm that 4. the individual plant of whole four genotype of selective polymerization A, B, C, D can obtain average grain length in F7 generations
Rice material (table 7)；
Grain length average of the table 7 in the individual plant of whole 4 genotype of F7 generations selective polymerization
| D309 | RM3585 | D463 | D755 | Grain length (mm) |
| Carry A | Carry B | Carry C | Carry D | 10.53 ± 0.82 (n=11) |
5. generation more than the paddy rice individual plant selfing on demand that will be selected in, to obtain the paddy rice strain that other economical characters are highly stable
System.
It can be seen that, be seeded in the same F2 colonies in May, 2011 from above example 1, example 2, example 3, is polymerize
In average grain length ＞ polymerization tetra- genotype of A, B, C, D of the individual plant of any 3 genotype in tetra- genotype of A, B, C, D
The average grain length ＞ of the individual plant of any 2 genotype carries the individual plant of any 1 genotype in tetra- genotype of A, B, C, D
Average grain length.It can be seen that, be seeded in the same F7 colonies in May, 2014 from example 4, example 5, example 6, example 7, gathered
Close any 3 bases in average grain length ＞ polymerization tetra- genotype of A, B, C, D of the individual plant in whole four genotype of A, B, C, D
Because of the average grain length of the individual plant of any 2 genotype in average grain length ＞ polymerization tetra- genotype of A, B, C, D of the individual plant of type
＞ carries the average grain length of the individual plant of any 1 genotype in tetra- genotype of A, B, C, D.If by different Fn colonies (n
To change numerical value) for example F2 colonies are compared with F7 colonies, then it is not the discussion scope of the present invention.
7 used leaf DNA extracting methods of examples of implementation PCR amplifications of the above use general CTAB methods；4 points
The PCR amplification programs of son mark for 94 DEG C 5 minutes, 94 DEG C of 35 circulations 30 seconds, 55 DEG C 30 seconds, 72 DEG C 1 minute, and last 72
DEG C 7 minutes；Pcr amplification product is detected using general denaturing polyacrylamide gel electrophoresis method and silver nitrate method staining.
Because these methods are all general normal experiment methods, therefore repeat no more herein.
Finally it is pointed out that the present invention is not limited only to the examples of implementation of the above, those skilled in the art are from this hair
Bright disclosure direct derivation or all flexible situations associated, are considered protection scope of the present invention.
Claims (6)
1. a kind of method of utilization molecular marking supplementary breeding length grain type paddy rice, it is characterised in that comprise the following steps：
(1) rice varieties Lemont and rice varieties are raised into n >=2 in rice No. 4 hybridization structure segregating populations Fn, the Fn；
(2) distinguished with 4 molecular labelings D309, RM3585, D463, D755 in the segregating population that PCR amplification steps (1) are obtained
The leaf DNA of individual plant；
(3) using the segregating population of 4 molecular labelings difference detecting step (1) description described in step (2), 4 and water are followed the trail of
The related quantitative trait locus position of grain of rice length；Wherein mark D309 is positioned at the chromosome of paddy rice the 3rd, for selecting to be located at paddy rice
The QTL relevant with grain length growth of 3rd chromosome：qGL-3-1YD；RM3585 is positioned at the chromosome of paddy rice the 3rd, for selecting position
In the QTL relevant with grain length growth of the chromosome of paddy rice the 3rd：qGL-3-2LE；D463 is positioned at the chromosome of paddy rice the 4th, for selecting
Select the QTL relevant with grain length growth positioned at the chromosome of paddy rice the 4th：qGL-4YD；D755 is positioned at the chromosome of paddy rice the 7th, is used for
With grain length growth relevant QTL of the selection positioned at the chromosome of paddy rice the 7th：qGL-7LE；D309 is expanded with raising No. 4 bands of rice
Type identical molecular marker gene type is named as A, RM3585 is expanded with Lemont banding pattern identical molecular marker genes
Type is named as B, and what D463 was expanded is named as C with raising No. 4 banding pattern identical molecular marker gene types of rice, and D755 amplifications are arrived
Be named as D with Lemont banding pattern identical molecular marker gene types；
(4) list for whole four genotype of A, B, C, D that polymerization procedure (3) is described in the segregating population that selection step (1) is described
Strain is to obtain the rice material of maximum grain length；Select step (1) description segregating population in polymerization procedure (3) description A, B, C,
The individual plant of any 3 genotype in tetra- genotype of D is to obtain the rice material that grain length takes second place；Select step (1) description
The individual plant of any 2 genotype in tetra- genotype of A, B, C, D that polymerization procedure (3) is described in segregating population is to obtain grain length
Relatively it polymerize the shorter rice material of the individual plant of any 3 genotype；Select to carry step in the segregating population of step (1) description
(3) individual plant of any 1 genotype in tetra- genotype of A, B, C, D of description relatively polymerize any 2 genes to obtain grain length
The shorter rice material of type；
(5) the individual plant selfing many generations for the different number of genotype for describing polymerization procedure (4), to obtain the long grain of different grain lengths
Type paddy rice homozygosis material.
2. the method for utilization molecular marking supplementary breeding length grain type paddy rice according to claim 1, it is characterised in that described
In step (1), Lemont is hybridized with raising rice No. 4, selection Lemont raises rice No. 4 as male parent, or selection as female parent
Rice No. 4 is raised as maternal Lemont as male parent to be hybridized.
3. the method for utilization molecular marking supplementary breeding length grain type paddy rice according to claim 1, it is characterised in that described
In step (3), molecular marker gene type A is the molecular labeling banding pattern for raising rice No. 4 detected using D309, but A is not
QGL-3-1YD genotype, due to D309 and qGL-3-1YD gene linkages, can be selected indirectly by selection A genotype
QGL-3-1YD genotype；Similarly can according to RM3585 and qGL-3-2LE gene linkages by indirect selections 1 B gene type come
Select qGL-3-2LE genotype；Selected according to D463 and qGL-4YD gene linkages by indirect selections C genotype
QGL-4YD genotype；Select qGL-7LE's by indirect selections D genotype according to D755 and qGL-7LE gene linkages
Genotype.
4. the method for utilization molecular marking supplementary breeding length grain type paddy rice according to claim 1, it is characterised in that described
In step (4), it polymerize the average grain length of individual plant of different number of genotype by being ordered as from big to small：It polymerize A, B, C, D complete
Any 2 ＞ comprise only in A, B, C, D any 1 in any 3 ＞ polymerizations A, B, C, D in 4, portion ＞ polymerizations A, B, C, D；Need
It is noted that above-mentioned sequence size is only applicable to be planted under same environmental condition to the segregating population in the same Fn generations
It is compared, n is fixed numbers in the Fn.
5. the method for utilization molecular marking supplementary breeding length grain type paddy rice according to claim 1, it is characterised in that described
In step (5), selfing algebraically should be actually needed determination according to breeding.
6. the method for utilization molecular marking supplementary breeding length grain type paddy rice according to claim 1, it is characterised in that long grain
Type paddy rice is defined as not including paddy rice of the non-groat length more than 9mm of awns.
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| CN108103230B (en) * | 2018-01-24 | 2021-03-23 | 中国水稻研究所 | Specific PCR molecular marker for detecting long and thin grain allele on rice grain shape QTLqGL35.1 |
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| CN110684858B (en) * | 2019-11-04 | 2023-09-26 | 广东省农业科学院水稻研究所 | Molecular marker of rice slender grain type gene and application thereof |
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| CN103289972A (en) * | 2012-03-02 | 2013-09-11 | 中国科学院上海生命科学研究院 | Rice long-grain related gene and application for same |
| CN103882145A (en) * | 2014-04-15 | 2014-06-25 | 江苏省农业科学院 | PCR (Polymerase Chain Reaction) molecular marking method for identifying allele mutation of rice long-grain gene qGL3 |
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| CN103289972A (en) * | 2012-03-02 | 2013-09-11 | 中国科学院上海生命科学研究院 | Rice long-grain related gene and application for same |
| CN103882145A (en) * | 2014-04-15 | 2014-06-25 | 江苏省农业科学院 | PCR (Polymerase Chain Reaction) molecular marking method for identifying allele mutation of rice long-grain gene qGL3 |
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