CN106755325B - A method of utilizing the medium grain weight type rice of molecular marking supplementary breeding - Google Patents
A method of utilizing the medium grain weight type rice of molecular marking supplementary breeding Download PDFInfo
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Abstract
The invention discloses a kind of methods using molecular marking supplementary breeding medium grain weight type rice: by Lemont and raise rice No. 4 and hybridize and construct segregating population, to the single plant in segregating population using 4 molecular labelings such as D140A, D456, D746, D1048 detect respectively 4 with grain weight related QTL:qGW-1YD, qGW-4LE, qGW-7LE, qGW-10LE.The molecular marker gene type identical with No. 4 banding patterns of rice are raised for marking D140A to amplify is named as A, the molecular marker gene type identical with Lemont banding pattern for marking D456, D746, D1048 to amplify respectively is respectively designated as B, C, D.The single plant of the different number of genes types in tetra- genotype of selective polymerization A, B, C, D is needed according to breeding to obtain the rice breed of different grain weights.The present invention is greatly improved the breeding efficiency that selection is carried out for grain principal characteristic shape.
Description
Technical field
The present invention relates to a kind of numbers for making rice grain weight reduction different come selective polymerization using molecular marker assisted selection
It measures character gene seat (QTL), in the method for breeding difference grain weight type rice.
Background technique
Rice is the important subsistence crop of China resident, and Rice Production is the very important composition portion of China's grain-production
Point.All the time, the raising of rice yield is the primary goal of China's Rice Production.The grain weight (grain weight) for improving rice can
To improve rice yield, therefore grain is the character that China rice breeding scientific research personnel extremely pays close attention to again.For a long time, rice
The conventional method of breeder's breeding difference grain weight type rice is carried out by Phenotypic Selection, just right after rice is mature
Grain is selected again.And using molecular labeling to rice grain weight related gene detect can rice seedling i.e. to grain weight into
Row selection, is greatly improved rice breeding efficiency.Currently, scientific research circle is it is known that the grain of rice is a typical quantity again
Character is controlled by multiple quantitative trait locus positions (QTL).Currently, rice scientific research circle has been oriented largely and rice grain weight
Relevant QTL.But how to reduce related QTL again by different in breeding work with grain and be aggregating use, with breeding
The rice material of different grain weights? in this context, the present invention proposes a kind of to polymerize 4 using molecular marker assisted selection and make
The QTL of rice grain weight reduction, in the method for breeding difference grain weight type rice.The present invention can compared to traditional Phenotypic Selection method
Breeding efficiency is greatly improved, breeding process is accelerated.
Summary of the invention
The present invention multiple reduces rice grain weight using selectively polymerizeing using molecular marker assisted selection
QTL, in the method for breeding medium grain weight type rice.Which, which polymerize, present invention mainly solves (a) reduces grain again
QTL? (b) selected using which molecular labeling? both of these problems.The present invention provides a kind of polymerization qGW-1YD, qGW-
4LE, qGW-7LE, qGW-10LE etc. 4 make the QTL of rice grain weight reduction, and utilize D140A, D456, D746, D1048 etc. 4
The method that molecular labeling carries out selection to this 4 QTL respectively.D140A, D456, D746, D1048 is effectively utilized in the present invention
It is chain with 4 QTL such as qGW-1YD, qGW-4LE, qGW-7LE, qGW-10LE respectively Deng 4 molecular labelings, and efficiently use
It is different to have reached accurate polymerization to the effectiveness of selection of this 4 QTL for 4 molecular labelings such as D140A, D456, D746, D1048
Grain weight QTL, with breeding difference grain weight type rice purpose.
The present invention the following steps are included:
(1) rice varieties Lemont and rice varieties rice No. 4 are raised to hybridize and construct segregating population Fn (n >=2)；
(2) segregating population obtained with 4 molecular labeling D140A, D456, D746, D1048 difference PCR amplification step (1)
The leaf DNA of interior different single plants；
(3) difference in the segregating population of 4 molecular labeling difference detecting step (1) descriptions described in step (2) is utilized
Single plant tracks 4 quantitative trait locus related with rice grain weight positions (QTL)；Wherein label D140A is positioned at rice the 1st
Chromosome weighs related QTL:qGW-1YD with grain on the 1st chromosome of rice for tracking；Label D456 is positioned at the dye of rice the 4th
Colour solid weighs related QTL:qGW-4LE with grain on the 4th chromosome of rice for tracking；Label D746 is positioned at the dyeing of rice the 7th
Body weighs related QTL:qGW-7LE with grain on the 7th chromosome of rice for tracking；Label D1048 is positioned at the dyeing of rice the 10th
Body weighs related QTL:qGW-10LE with grain on the 10th chromosome of rice for tracking；By mark D140A amplify with raise rice
The identical molecular marker gene type of No. 4 banding patterns is named as A, the molecule mark identical with Lemont banding pattern that D456 will be marked to amplify
Note genotype is named as B, and the molecular marker gene type identical with Lemont banding pattern for marking D746 to amplify is named as C, will
The molecular marker gene type identical with Lemont banding pattern that label D1048 is amplified is named as D；
(4) whole 4 genotype of A, B, C, D of polymerization procedure (3) description in the segregating population of selection step (1) description
Single plant is to obtain the smallest rice material of average mass of 1000 kernel；Select polymerization procedure (3) description in the segregating population of step (1) description
Tetra- genotype of A, B, C, D in any 3 genotype single plant to obtain average mass of 1000 kernel than polymerization A, B, C, D all 4
The bigger rice material of the single plant of a genotype；Select step (1) description segregating population in polymerization procedure (3) description A, B,
C, the single plant of any 2 genotype in tetra- genotype of D is relatively polymerize in tetra- genotype of A, B, C, D with obtaining average mass of 1000 kernel
Any 3 genotype the bigger rice material of single plant；Step (3) are carried in the segregating population of selection step (1) description to retouch
The single plant of any 1 genotype in tetra- genotype of A, B, C, the D stated relatively polymerize A, B, C, D tetra- to obtain average mass of 1000 kernel
The bigger rice material of the single plant of any 2 genotype in genotype；
(5) the individual plant selfing mostly generation of the different number of genotype of step (4) description will be carried, to obtain different mass of 1000 kernel
Rice homozygosis material.
Further, in the step (1), Lemont is hybridized with raising rice No. 4, Lemont may be selected as maternal
Rice No. 4 are raised as male parent, also be can choose and raised rice No. 4 and hybridized as female parent Lemont as male parent.
Further, in the step (2), using 4 molecular labelings D140A, D456, D746, D1048 come PCR amplification
The number of single plant in segregating population should be actually needed according to breeding and determine；By taking F2 group as an example, to be screened in step (4)
1 plant of single plant for carrying whole 4 genotype of A, B, C, D is obtained, no less than 256 single plant (calculation methods 4 should be theoretically detected
× 4 × 4 × 4=256), in another example, to obtain 1 plant of 3 base carried in tetra- genotype of A, B, C, D in step (4) screening
Because of the single plant of type, it should theoretically detect no less than 64 single plants (calculation method is 4 × 4 × 4=64)；Certainly, example above
It is theoretic supposition, during atual detection, it should determine the single plant number of detection as needed；Generally speaking, it detects
Single plant number it is more, then the chance for obtaining the different desired single plants of step (4) description is just bigger.
Further, in the step (3), the banding pattern of molecular marker gene type A is raising rice No. 4 with using label D140A
The banding pattern that the denaturing polyacrylamide gel electrophoresis that upper PCR amplification passes through 8% concentration again detects is identical, but A is not qGW-
The genotype of 1YD can select qGW-1YD's indirectly by the genotype of selection A due to D140A and qGW-1YD gene linkage
Genotype；It similarly can be chain and the genotype of qGW-4LE is selected by indirect selections 1 B gene type according to D456 and qGW-4LE；
It is chain and the genotype of qGW-7LE is selected by indirect selections C genotype according to D746 and qGW-7LE；According to D1048 with
QGW-10LE is chain and the genotype of qGW-10LE is selected by indirect selections D genotype.
Further, in the step (4), polymerize the average mass of 1000 kernel of the single plant of different number of genotype by from it is small to
Big sequence are as follows: any 3 genotype < in whole 4 genotype < polymerization A, B, C, the D of polymerization A, B, C, D polymerize A, B, C, D
In any 2 genotype < carry A, B, C, D in any 1 genotype；It should be pointed out that above-mentioned sequence size is only fitted
For be planted in the single plant of the segregating population under the same environmental condition to same Fn (n >=2 and n be fixed numbers) in the generation into
Row compares.
Further, in the step (5), selfing algebra should be actually needed according to breeding and determine.
Further, medium grain weight type rice is defined as being planted in the non-decladding after the mature harvest of Hangzhou or Lingshui
Grain dry after rice of the mass of 1000 kernel between 21 grams to 26.5 grams.
Specific embodiment
Embodiment 1: selection carries the single plant of any one genotype in tetra- genotype of A, B, C, D to obtain average thousand
Grain again the F2 less than 25.48 grams (average mass of 1000 kernel between 25.48 grams and 24.42 grams) for rice material
1. rice varieties Lemont is made female parent, rice No. 4 are raised with rice varieties and is hybridized and constructed F2 generation point as male parent
Peeling off, (Lemont is the japonica rice variety from the U.S. to body, raises rice No. 4 as the rice variety from Jiangsu, Lemont and raises rice No. 4
It is provided by country, China Paddy Rice Inst Rice Germplasm Resources mid-term library).
2. with 191 lists in 4 molecular labeling D140A, D456, D746, D1048 difference PCR amplification F2 segregating populations
Strain, this F2 segregating population including 191 single plants are sowed in May, 2011 in the rice in China research of Hangzhou Fuyang area
Tested field；The sequence of this 4 molecular labelings of D140A, D456, D746, D1048 is shown in Zeng Y.X., et
al.Development of 1047 insertion-deletion markers for rice genetic studies
And breeding.Genetics and Molecular Research, 2013, volume 12: 5226-5235.
3. tracking A genotype by label D140A to track from qGW-1YD gene (point of A genotype for raising rice No. 4
Sub- marker bands are the banding pattern that D140A label amplification raises rice No. 4)；1 B gene type is tracked by label D456 to come to track
The qGW-4LE gene (banding pattern that the molecular labeling banding pattern of 1 B gene type is D456 label amplification Lemont) of Lemont；Pass through label
D746 tracks C genotype, and to track the qGW-7LE gene from Lemont, (the molecular labeling banding pattern of C genotype is D746 label
Expand the banding pattern of Lemont)；D genotype is tracked by label D1048 to track qGW-10LE gene (the D base from Lemont
Because of the banding pattern that the molecular labeling banding pattern of type is D1048 label amplification Lemont).
4. the single plant that selection carries any one genotype in tetra- genotype of A, B, C, D is small to obtain average mass of 1000 kernel
In the rice material (table 1) of 25.48 grams (average mass of 1000 kernel is between 25.48 grams and 24.42 grams).
Table 1. is utilized respectively the list for the carrying different genotype that 4 different molecular labelings are selected in F2 from generation to generation
The average mass of 1000 kernel of strain, the F2 group are seeded in Hangzhou Fuyang China Paddy Rice Inst in May, 2011
Embodiment 2: the single plant of any 2 genotype in tetra- genotype of selective polymerization A, B, C, D is to obtain average thousand
Grain again the F2 less than 24.98 grams (average mass of 1000 kernel between 24.98 grams and 22.87 grams) for rice material
1. rice varieties Lemont is made female parent, rice No. 4 are raised with rice varieties and is hybridized and constructed F2 generation point as male parent
Peeling off, (Lemont is the japonica rice variety from the U.S. to body, raises rice No. 4 as the rice variety from Jiangsu, Lemont and raises rice No. 4
It is provided by country, China Paddy Rice Inst Rice Germplasm Resources mid-term library).
2. with 191 lists in 4 molecular labeling D140A, D456, D746, D1048 difference PCR amplification F2 segregating populations
Strain, this F2 segregating population including 191 single plants are sowed in May, 2011 in the rice in China research of Hangzhou Fuyang area
Tested field；The sequence of this 4 molecular labelings of D140A, D456, D746, D1048 is shown in Zeng Y.X., et
al.Development of 1047 insertion-deletion markers for rice genetic studies
And breeding.Genetics and Molecular Research, 2013, volume 12: 5226-5235.
3. tracking A genotype by label D140A to track from qGW-1YD gene (point of A genotype for raising rice No. 4
Sub- marker bands are the banding pattern that D140A label amplification raises rice No. 4)；1 B gene type is tracked by label D456 to come to track
The qGW-4LE gene (banding pattern that the molecular labeling banding pattern of 1 B gene type is D456 label amplification Lemont) of Lemont；Pass through label
D746 tracks C genotype, and to track the qGW-7LE gene from Lemont, (the molecular labeling banding pattern of C genotype is D746 label
Expand the banding pattern of Lemont)；D genotype is tracked by label D1048 to track qGW-10LE gene (the D base from Lemont
Because of the banding pattern that the molecular labeling banding pattern of type is D1048 label amplification Lemont).
4. the single plant of any 2 genotype in tetra- genotype of selective polymerization A, B, C, D is small to obtain average mass of 1000 kernel
In the rice material (table 2) of 24.98 grams (average mass of 1000 kernel is between 24.98 grams and 22.87 grams).
Table 2. utilizes any 2 genotype in 4 Marker-assisted selection polymerization tetra- genotype of A, B, C, D in F2 from generation to generation
Single plant average mass of 1000 kernel, which is seeded in Hangzhou Fuyang China Paddy Rice Inst in May, 2011
Embodiment 3: the single plant of any 3 genotype in tetra- genotype of selective polymerization A, B, C, D is to obtain average thousand
Grain again the F2 less than 24.25 grams (average mass of 1000 kernel between 24.25 grams and 21.43 grams) for rice material
1. rice varieties Lemont is made female parent, rice No. 4 are raised with rice varieties and is hybridized and constructed F2 generation point as male parent
Peeling off, (Lemont is the japonica rice variety from the U.S. to body, raises rice No. 4 as the rice variety from Jiangsu, Lemont and raises rice No. 4
It is provided by country, China Paddy Rice Inst Rice Germplasm Resources mid-term library).
2. with 191 lists in 4 molecular labeling D140A, D456, D746, D1048 difference PCR amplification F2 segregating populations
Strain, this F2 segregating population including 191 single plants are sowed in May, 2011 in the rice in China research of Hangzhou Fuyang area
Tested field；The sequence of this 4 molecular labelings of D140A, D456, D746, D1048 is shown in Zeng Y.X., et
al.Development of 1047 insertion-deletion markers for rice genetic studies
And breeding.Genetics and Molecular Research, 2013, volume 12: 5226-5235.
3. tracking A genotype by label D140A to track from qGW-1YD gene (point of A genotype for raising rice No. 4
Sub- marker bands are the banding pattern that D140A label amplification raises rice No. 4)；1 B gene type is tracked by label D456 to come to track
The qGW-4LE gene (banding pattern that the molecular labeling banding pattern of 1 B gene type is D456 label amplification Lemont) of Lemont；Pass through label
D746 tracks C genotype, and to track the qGW-7LE gene from Lemont, (the molecular labeling banding pattern of C genotype is D746 label
Expand the banding pattern of Lemont)；D genotype is tracked by label D1048 to track qGW-10LE gene (the D base from Lemont
Because of the banding pattern that the molecular labeling banding pattern of type is D1048 label amplification Lemont).
4. the single plant of any 3 genotype in tetra- genotype of selective polymerization A, B, C, D is small to obtain average mass of 1000 kernel
In the rice material (table 3) of 24.25 grams (average mass of 1000 kernel is between 24.25 grams and 21.43 grams).
Table 3. utilizes any 3 genotype in 4 Marker-assisted selection polymerization tetra- genotype of A, B, C, D in F2 from generation to generation
Single plant average mass of 1000 kernel, which is seeded in Hangzhou Fuyang China Paddy Rice Inst in May, 2011
Embodiment 4: selective polymerization A, B, C, D all 4 genotype single plant with obtain average mass of 1000 kernel than carry A, B,
C, the smaller F2 of average mass of 1000 kernel of the single plant of any 3 genotype in tetra- genotype of D is for rice material
1. rice varieties Lemont is made female parent, rice No. 4 are raised with rice varieties and is hybridized and constructed F2 generation point as male parent
Peeling off, (Lemont is the japonica rice variety from the U.S. to body, raises rice No. 4 as the rice variety from Jiangsu, Lemont and raises rice No. 4
It is provided by country, China Paddy Rice Inst Rice Germplasm Resources mid-term library).
2. with 4 molecular labeling D140A, D456, D746, D1048 difference PCR amplification F2 for 191 in segregating population
Single plant, this F2 including 191 single plants are sowed in May, 2011 in Hangzhou Fuyang area rice in China for segregating population
Research institute test site；The sequence of this 4 molecular labelings of D140A, D456, D746, D1048 is shown in Zeng Y.X., et
al.Development of 1047 insertion-deletion markers for rice genetic studies
And breeding.Genetics and Molecular Research, 2013, volume 12: 5226-5235.
3. tracking A genotype by label D140A to track from qGW-1YD gene (point of A genotype for raising rice No. 4
Sub- marker bands are the banding pattern that D140A label amplification raises rice No. 4)；1 B gene type is tracked by label D456 to come to track
The qGW-4LE gene (banding pattern that the molecular labeling banding pattern of 1 B gene type is D456 label amplification Lemont) of Lemont；Pass through label
D746 tracks C genotype, and to track the qGW-7LE gene from Lemont, (the molecular labeling banding pattern of C genotype is D746 label
Expand the banding pattern of Lemont)；D genotype is tracked by label D1048 to track qGW-10LE gene (the D base from Lemont
Because of the banding pattern that the molecular labeling banding pattern of type is D1048 label amplification Lemont).
4. selecting the single plant for carrying whole 4 genotype of A, B, C, D to obtain mass of 1000 kernel as 21.43 grams of rice material
(table 4).
Table 4. utilizes thousand of the single plant of 4 whole four genotype of Marker-assisted selection polymerization A, B, C, D in F2 from generation to generation
Weight, the F2 group are seeded in Hangzhou Fuyang China Paddy Rice Inst in May, 2011
Embodiment 5: selection carries the single plant of any 1 genotype in tetra- genotype of A, B, C, D to obtain average thousand
Grain again the F7 less than 26.14 grams (average mass of 1000 kernel between 26.14 grams and 25.83 grams) for rice material
1. rice varieties Lemont is made female parent, rice No. 4 are raised with rice varieties and is hybridized and constructed F7 generation point as male parent
Peeling off, (Lemont is the japonica rice variety from the U.S. to body, raises rice No. 4 as the rice variety from Jiangsu, Lemont and raises rice No. 4
It is provided by country, China Paddy Rice Inst Rice Germplasm Resources mid-term library).
2. with 4 molecular labeling D140A, D456, D746, D1048 difference PCR amplification F7 for 221 in segregating population
Single plant, this F7 including 221 single plants are sowed in May, 2014 in Hangzhou Fuyang area rice in China for segregating population
Research institute test site；The sequence of this 4 molecular labelings of D140A, D456, D746, D1048 is shown in Zeng Y.X., et
al.Development of 1047 insertion-deletion markers for rice genetic studies
And breeding.Genetics and Molecular Research, 2013, volume 12: 5226-5235.
3. tracking A genotype by label D140A to track from qGW-1YD gene (point of A genotype for raising rice No. 4
Sub- marker bands are the banding pattern that D140A label amplification raises rice No. 4)；1 B gene type is tracked by label D456 to come to track
The qGW-4LE gene (banding pattern that the molecular labeling banding pattern of 1 B gene type is D456 label amplification Lemont) of Lemont；Pass through label
D746 tracks C genotype, and to track the qGW-7LE gene from Lemont, (the molecular labeling banding pattern of C genotype is D746 label
Expand the banding pattern of Lemont)；D genotype is tracked by label D1048 to track qGW-10LE gene (the D base from Lemont
Because of the banding pattern that the molecular labeling banding pattern of type is D1048 label amplification Lemont).
4. the single plant that selection carries any 1 genotype in tetra- genotype of A, B, C, D is small to obtain average mass of 1000 kernel
In the rice material (table 5) of 26.14 grams (average mass of 1000 kernel is between 26.14 grams and 25.83 grams).
Table 5. is utilized respectively the list for the carrying different genotype that 4 different molecular labelings are selected in F7 from generation to generation
The average mass of 1000 kernel of strain, the F7 group are seeded in Hangzhou Fuyang China Paddy Rice Inst in May, 2014
Embodiment 6: the single plant of any 2 genotype in tetra- genotype of selective polymerization A, B, C, D is to obtain average thousand
Grain again the F7 less than 25.43 grams (average mass of 1000 kernel between 25.43 grams and 24.42 grams) for rice material
1. rice varieties Lemont is made female parent, rice No. 4 are raised with rice varieties and is hybridized and constructed F7 generation point as male parent
Peeling off, (Lemont is the japonica rice variety from the U.S. to body, raises rice No. 4 as the rice variety from Jiangsu, Lemont and raises rice No. 4
It is provided by country, China Paddy Rice Inst Rice Germplasm Resources mid-term library).
2. with 4 molecular labeling D140A, D456, D746, D1048 difference PCR amplification F7 for 221 in segregating population
Single plant, this F7 including 221 single plants are sowed in May, 2014 in Hangzhou Fuyang area rice in China for segregating population
Research institute test site；The sequence of this 4 molecular labelings of D140A, D456, D746, D1048 is shown in Zeng Y.X., et
al.Development of 1047 insertion-deletion markers for rice genetic studies
And breeding.Genetics and Molecular Research, 2013, volume 12: 5226-5235.
3. tracking A genotype by label D140A to track from qGW-1YD gene (point of A genotype for raising rice No. 4
Sub- marker bands are the banding pattern that D140A label amplification raises rice No. 4)；1 B gene type is tracked by label D456 to come to track
The qGW-4LE gene (banding pattern that the molecular labeling banding pattern of 1 B gene type is D456 label amplification Lemont) of Lemont；Pass through label
D746 tracks C genotype, and to track the qGW-7LE gene from Lemont, (the molecular labeling banding pattern of C genotype is D746 label
Expand the banding pattern of Lemont)；D genotype is tracked by label D1048 to track qGW-10LE gene (the D base from Lemont
Because of the banding pattern that the molecular labeling banding pattern of type is D1048 label amplification Lemont).
4. the single plant of any 2 genotype in tetra- genotype of selective polymerization A, B, C, D is small to obtain average mass of 1000 kernel
In the rice material (table 6) of 25.43 grams (average mass of 1000 kernel is between 25.43 grams and 24.42 grams).
Table 6. utilizes any 2 genotype in 4 Marker-assisted selection polymerization tetra- genotype of A, B, C, D in F7 from generation to generation
Single plant average mass of 1000 kernel, which is seeded in Hangzhou Fuyang China Paddy Rice Inst in May, 2014
Embodiment 7: the single plant of any 3 genotype in tetra- genotype of selective polymerization A, B, C, D is to obtain average thousand
Grain again the F7 less than 24.80 grams (average mass of 1000 kernel between 24.80 grams and 23.31 grams) for rice material
1. rice varieties Lemont is made female parent, rice No. 4 are raised with rice varieties and is hybridized and constructed F7 generation point as male parent
Peeling off, (Lemont is the japonica rice variety from the U.S. to body, raises rice No. 4 as the rice variety from Jiangsu, Lemont and raises rice No. 4
It is provided by country, China Paddy Rice Inst Rice Germplasm Resources mid-term library).
2. with 4 molecular labeling D140A, D456, D746, D1048 difference PCR amplification F7 for 221 in segregating population
Single plant, this F7 including 221 single plants are sowed in May, 2014 in Hangzhou Fuyang area rice in China for segregating population
Research institute test site；The sequence of this 4 molecular labelings of D140A, D456, D746, D1048 is shown in Zeng Y.X., et
al.Development of 1047 insertion-deletion markers for rice genetic studies
And breeding.Genetics and Molecular Research, 2013, volume 12: 5226-5235.
3. tracking A genotype by label D140A to track from qGW-1YD gene (point of A genotype for raising rice No. 4
Sub- marker bands are the banding pattern that D140A label amplification raises rice No. 4)；1 B gene type is tracked by label D456 to come to track
The qGW-4LE gene (banding pattern that the molecular labeling banding pattern of 1 B gene type is D456 label amplification Lemont) of Lemont；Pass through label
D746 tracks C genotype, and to track the qGW-7LE gene from Lemont, (the molecular labeling banding pattern of C genotype is D746 label
Expand the banding pattern of Lemont)；D genotype is tracked by label D1048 to track qGW-10LE gene (the D base from Lemont
Because of the banding pattern that the molecular labeling banding pattern of type is D1048 label amplification Lemont).
4. the single plant of any 3 genotype in tetra- genotype of selective polymerization A, B, C, D is small to obtain average mass of 1000 kernel
In the rice material (table 7) of 24.80 grams (average mass of 1000 kernel is between 24.80 grams and 23.31 grams).
Table 7. utilizes any 3 genotype in 4 Marker-assisted selection polymerization tetra- genotype of A, B, C, D in F7 from generation to generation
Single plant average mass of 1000 kernel, which is seeded in Hangzhou Fuyang China Paddy Rice Inst in May, 2014
Embodiment 8: the single plant of whole four genotype of selective polymerization A, B, C, D is to obtain average mass of 1000 kernel as 21.63 grams
F7 for rice material
1. rice varieties Lemont is made female parent, rice No. 4 are raised with rice varieties and is hybridized and constructed F7 generation point as male parent
Peeling off, (Lemont is the japonica rice variety from the U.S. to body, raises rice No. 4 as the rice variety from Jiangsu, Lemont and raises rice No. 4
It is provided by country, China Paddy Rice Inst Rice Germplasm Resources mid-term library).
2. with 4 molecular labeling D140A, D456, D746, D1048 difference PCR amplification F7 for 221 in segregating population
Single plant, this F7 including 221 single plants are sowed in May, 2014 in Hangzhou Fuyang area rice in China for segregating population
Research institute test site；The sequence of this 4 molecular labelings of D140A, D456, D746, D1048 is shown in Zeng Y.X., et
al.Development of 1047 insertion-deletion markers for rice genetic studies
And breeding.Genetics and Molecular Research, 2013, volume 12: 5226-5235.
3. tracking A genotype by label D140A to track from qGW-1YD gene (point of A genotype for raising rice No. 4
Sub- marker bands are the banding pattern that D140A label amplification raises rice No. 4)；1 B gene type is tracked by label D456 to come to track
The qGW-4LE gene (banding pattern that the molecular labeling banding pattern of 1 B gene type is D456 label amplification Lemont) of Lemont；Pass through label
D746 tracks C genotype, and to track the qGW-7LE gene from Lemont, (the molecular labeling banding pattern of C genotype is D746 label
Expand the banding pattern of Lemont)；D genotype is tracked by label D1048 to track qGW-10LE gene (the D base from Lemont
Because of the banding pattern that the molecular labeling banding pattern of type is D1048 label amplification Lemont).
4. the single plant of whole 4 genotype of selective polymerization A, B, C, D is to obtain the rice material that average mass of 1000 kernel is 21.63 grams
Expect (table 8).
Table 8. utilizes average the thousand of the single plant of 4 whole 4 genotype of Marker-assisted selection polymerization A, B, C, D in F7 from generation to generation
Grain weight, the F7 group are seeded in Hangzhou Fuyang China Paddy Rice Inst in May, 2014
Embodiment 9: selection carries the single plant of any 1 genotype in tetra- genotype of A, B, C, D to obtain average thousand
Grain again the F10 less than 26.07 grams (average mass of 1000 kernel between 26.07 grams and 25.22 grams) for rice material
1. rice varieties Lemont is made female parent, rice No. 4 are raised with rice varieties and is hybridized and constructed F10 generation point as male parent
Peeling off, (Lemont is the japonica rice variety from the U.S. to body, raises rice No. 4 as the rice variety from Jiangsu, Lemont and raises rice No. 4
It is provided by country, China Paddy Rice Inst Rice Germplasm Resources mid-term library).
2. with 4 molecular labeling D140A, D456, D746, D1048 difference PCR amplification F10 for 221 in segregating population
Single plant, this F10 including 221 single plants are sowed in November, 2015 in Hainan Province's Lingshui county China Water for segregating population
The numerous test center in rice research institute south；The sequence of this 4 molecular labelings of D140A, D456, D746, D1048 is shown in Zeng Y.X., et
al.Development of 1047 insertion-deletion markers for rice genetic studies
And breeding.Genetics and Molecular Research, 2013, volume 12: 5226-5235.
3. tracking A genotype by label D140A to track from qGW-1YD gene (point of A genotype for raising rice No. 4
Sub- marker bands are the banding pattern that D140A label amplification raises rice No. 4)；1 B gene type is tracked by label D456 to come to track
The qGW-4LE gene (banding pattern that the molecular labeling banding pattern of 1 B gene type is D456 label amplification Lemont) of Lemont；Pass through label
D746 tracks C genotype, and to track the qGW-7LE gene from Lemont, (the molecular labeling banding pattern of C genotype is D746 label
Expand the banding pattern of Lemont)；D genotype is tracked by label D1048 to track qGW-10LE gene (the D base from Lemont
Because of the banding pattern that the molecular labeling banding pattern of type is D1048 label amplification Lemont).
4. the single plant that selection carries any 1 genotype in tetra- genotype of A, B, C, D is small to obtain average mass of 1000 kernel
In the rice material (table 9) of 26.07 grams (average mass of 1000 kernel is between 26.07 grams and 25.22 grams).
Table 9. is utilized respectively the carrying different genotype that 4 different molecular labelings are selected in F10 from generation to generation
The average mass of 1000 kernel of single plant, the F10 group were seeded in the numerous test in south of Hainan Lingshui China Paddy Rice Inst in November, 2015
The heart
Embodiment 10: the single plant of any 2 genotype in tetra- genotype of selective polymerization A, B, C, D is to obtain average thousand
Grain again the F10 less than 25.57 grams (average mass of 1000 kernel between 25.57 grams and 24.35 grams) for rice material
1. rice varieties Lemont is made female parent, rice No. 4 are raised with rice varieties and is hybridized and constructed F10 generation point as male parent
Peeling off, (Lemont is the japonica rice variety from the U.S. to body, raises rice No. 4 as the rice variety from Jiangsu, Lemont and raises rice No. 4
It is provided by country, China Paddy Rice Inst Rice Germplasm Resources mid-term library).
2. with 4 molecular labeling D140A, D456, D746, D1048 difference PCR amplification F10 for 221 in segregating population
Single plant, this F10 including 221 single plants are sowed in November, 2015 in Hainan Province's Lingshui county China Water for segregating population
The numerous test center in rice research institute south；The sequence of this 4 molecular labelings of D140A, D456, D746, D1048 is shown in Zeng Y.X., et
al.Development of 1047 insertion-deletion markers for rice genetic studies
And breeding.Genetics and Molecular Research, 2013, volume 12: 5226-5235.
3. tracking A genotype by label D140A to track from qGW-1YD gene (point of A genotype for raising rice No. 4
Sub- marker bands are the banding pattern that D140A label amplification raises rice No. 4)；1 B gene type is tracked by label D456 to come to track
The qGW-4LE gene (banding pattern that the molecular labeling banding pattern of 1 B gene type is D456 label amplification Lemont) of Lemont；Pass through label
D746 tracks C genotype, and to track the qGW-7LE gene from Lemont, (the molecular labeling banding pattern of C genotype is D746 label
Expand the banding pattern of Lemont)；D genotype is tracked by label D1048 to track qGW-10LE gene (the D base from Lemont
Because of the banding pattern that the molecular labeling banding pattern of type is D1048 label amplification Lemont).
4. the single plant of any 2 genotype in tetra- genotype of selective polymerization A, B, C, D is small to obtain average mass of 1000 kernel
In the rice material (table 10) of 25.57 grams (average mass of 1000 kernel is between 25.57 grams and 24.35 grams).
Table 10. utilizes any 2 genes in 4 Marker-assisted selection polymerization tetra- genotype of A, B, C, D in F10 from generation to generation
The average mass of 1000 kernel of the single plant of type, the F10 group are seeded in the numerous examination in south of Hainan Lingshui China Paddy Rice Inst in November, 2015
Test center
Embodiment 11: the single plant of any 3 genotype in tetra- genotype of selective polymerization A, B, C, D is to obtain average thousand
Grain again the F10 less than 25.17 grams (average mass of 1000 kernel between 25.17 grams and 23.30 grams) for rice material
1. rice varieties Lemont is made female parent, rice No. 4 are raised with rice varieties and is hybridized and constructed F10 generation point as male parent
Peeling off, (Lemont is the japonica rice variety from the U.S. to body, raises rice No. 4 as the rice variety from Jiangsu, Lemont and raises rice No. 4
It is provided by country, China Paddy Rice Inst Rice Germplasm Resources mid-term library).
2. with 4 molecular labeling D140A, D456, D746, D1048 difference PCR amplification F10 for 221 in segregating population
Single plant, this F10 including 221 single plants are sowed in November, 2015 in Hainan Province's Lingshui county China Water for segregating population
The numerous test center in rice research institute south；The sequence of this 4 molecular labelings of D140A, D456, D746, D1048 is shown in Zeng Y.X., et
al.Development of 1047 insertion-deletion markers for rice genetic studies
And breeding.Genetics and Molecular Research, 2013, volume 12: 5226-5235.
3. tracking A genotype by label D140A to track from qGW-1YD gene (point of A genotype for raising rice No. 4
Sub- marker bands are the banding pattern that D140A label amplification raises rice No. 4)；1 B gene type is tracked by label D456 to come to track
The qGW-4LE gene (banding pattern that the molecular labeling banding pattern of 1 B gene type is D456 label amplification Lemont) of Lemont；Pass through label
D746 tracks C genotype, and to track the qGW-7LE gene from Lemont, (the molecular labeling banding pattern of C genotype is D746 label
Expand the banding pattern of Lemont)；D genotype is tracked by label D1048 to track qGW-10LE gene (the D base from Lemont
Because of the banding pattern that the molecular labeling banding pattern of type is D1048 label amplification Lemont).
4. the single plant of any 3 genotype in tetra- genotype of selective polymerization A, B, C, D is small to obtain average mass of 1000 kernel
In the rice material (table 11) of 25.17 grams (average mass of 1000 kernel is between 25.17 grams and 23.30 grams).
Table 11. utilizes any 3 genes in 4 Marker-assisted selection polymerization tetra- genotype of A, B, C, D in F10 from generation to generation
The average mass of 1000 kernel of the single plant of type, the F10 group are seeded in the numerous examination in south of Hainan Lingshui China Paddy Rice Inst in November, 2015
Test center
Embodiment 12: the single plant of whole 4 genotype of selective polymerization A, B, C, D is to obtain average mass of 1000 kernel as 23.84 grams
F10 for rice material
1. rice varieties Lemont is made female parent, rice No. 4 are raised with rice varieties and is hybridized and constructed F10 generation point as male parent
Peeling off, (Lemont is the japonica rice variety from the U.S. to body, raises rice No. 4 as the rice variety from Jiangsu, Lemont and raises rice No. 4
It is provided by country, China Paddy Rice Inst Rice Germplasm Resources mid-term library).
2. with 4 molecular labeling D140A, D456, D746, D1048 difference PCR amplification F10 for 221 in segregating population
Single plant, this F10 including 221 single plants are sowed in November, 2015 in Hainan Province's Lingshui county China Water for segregating population
The numerous test center in rice research institute south；The sequence of this 4 molecular labelings of D140A, D456, D746, D1048 is shown in Zeng Y.X., et
al.Development of 1047 insertion-deletion markers for rice genetic studies
And breeding.Genetics and Molecular Research, 2013, volume 12: 5226-5235.
3. tracking A genotype by label D140A to track from qGW-1YD gene (point of A genotype for raising rice No. 4
Sub- marker bands are the banding pattern that D140A label amplification raises rice No. 4)；1 B gene type is tracked by label D456 to come to track
The qGW-4LE gene (banding pattern that the molecular labeling banding pattern of 1 B gene type is D456 label amplification Lemont) of Lemont；Pass through label
D746 tracks C genotype, and to track the qGW-7LE gene from Lemont, (the molecular labeling banding pattern of C genotype is D746 label
Expand the banding pattern of Lemont)；D genotype is tracked by label D1048 to track qGW-10LE gene (the D base from Lemont
Because of the banding pattern that the molecular labeling banding pattern of type is D1048 label amplification Lemont).
4. the single plant of whole 4 genotype of selective polymerization A, B, C, D is to obtain the rice material that average mass of 1000 kernel is 23.84 grams
Expect (table 12).
Table 12. utilizes being averaged for the single plant of 4 whole 4 genotype of Marker-assisted selection polymerization A, B, C, D in F10 from generation to generation
Mass of 1000 kernel, the F10 group are seeded in Hainan Lingshui China Paddy Rice Inst numerous test center in south in November, 2015
From above embodiments 1, embodiment 2, embodiment 3, embodiment 4 as can be seen that being seeded in Hangzhou in May, 2011
In the same F2 group, average mass of 1000 kernel (for 21.43 grams) the < carrying A of the single plant of whole four genotype of polymerization A, B, C, D,
B, the average mass of 1000 kernel of the single plant of any 3 genotype in tetra- genotype of C, D (between 24.25 grams and 21.43 grams)
< carries the average mass of 1000 kernel of the single plant of any 2 genotype in tetra- genotype of A, B, C, D (between 24.98 grams and 22.87
Between gram) < carry tetra- genotype of A, B, C, D in any 1 genotype single plant average mass of 1000 kernel (between 25.48 grams
And between 24.42 grams).
From above embodiments 5, embodiment 6, embodiment 7, embodiment 8 as can be seen that being seeded in Hangzhou in May, 2014
In the same F7 group, average mass of 1000 kernel (for 21.63 grams) the < carrying A of the single plant of whole four genotype of polymerization A, B, C, D,
B, the average mass of 1000 kernel of the single plant of any 3 genotype in tetra- genotype of C, D (between 24.80 grams and 23.31 grams)
< carries the average mass of 1000 kernel of the single plant of any 2 genotype in tetra- genotype of A, B, C, D (between 25.43 grams and 24.42
Between gram) < carry tetra- genotype of A, B, C, D in any 1 genotype single plant average mass of 1000 kernel (between 26.14 grams
And between 25.83 grams).
From above embodiments 9, embodiment 10, embodiment 11, embodiment 12 as can be seen that being seeded in sea in November, 2015
In the same F10 group in south, average mass of 1000 kernel (for 23.84 grams) < of the single plant of whole four genotype of polymerization A, B, C, D
The average mass of 1000 kernel of the single plant of any 3 genotype in tetra- genotype of A, B, C, D is carried (between 25.17 grams and 23.30 grams
Between) < carry any 2 genotype in tetra- genotype of A, B, C, D single plant average mass of 1000 kernel (between 25.57 grams and
Between 24.35 grams) < carry tetra- genotype of A, B, C, D in any 1 genotype single plant average mass of 1000 kernel (between
Between 26.07 grams and 25.22 grams).
Generally speaking, in the same Fn (n >=2 and n the is fixed numbers) segregating population being in the same localities to sowing
For single plant, the more single plant of the number of A, B, C, D different genotype is carried, then the average mass of 1000 kernel of these single plants is smaller；It takes
The fewer single plant of number with A, B, C, D different genotype, then the average mass of 1000 kernel of these single plants is bigger.It should be noted that
The rule is only applicable to the single plant being planted in the same Fn (n >=2 and n the is fixed numbers) segregating population in same place, such as
Single plant in different Fn (n >=2 and n is fixed numbers) group is compared by fruit, such as by the single plant of F2 group and F7 group
Single plant be compared, then be not discussion range of the invention.
Experimental method mentioned by the present invention is summarized as follows: the extracting method of leaf DNA used in PCR amplification is using logical
CTAB method；The PCR amplification program of 4 molecular labelings be 94 DEG C 5 minutes, 35 circulation 94 DEG C 30 seconds, 55 DEG C 30 seconds,
72 DEG C 1 minute, and it is last 72 DEG C 7 minutes；Pcr amplification product uses the denaturing polyacrylamide gel of 8% general concentration
Electrophoresis and silver nitrate method staining are detected.Since these methods are all general routine experiment methods, herein not
It repeats again.
Finally it is pointed out that the present invention be not limited only to more than examples of implementation, 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 (7)
1. a kind of method using the medium grain weight type rice of molecular marking supplementary breeding, which comprises the following steps:
(1) rice varieties Lemont and rice varieties rice No. 4 are raised to hybridize and construct segregating population Fn (n >=2)；
(2) in the segregating population with 4 molecular labeling D140A, D456, D746, D1048 difference PCR amplification step (1) acquisitions
The leaf DNA of different single plants；
(3) the different single plants in the segregating population of detecting step (1) description are distinguished using 4 molecular labelings described in step (2),
Track 4 quantitative trait locus related with rice grain weight positions (QTL)；Wherein label D140A is positioned at the dyeing of rice the 1st
Body weighs related QTL:qGW-1YD with grain on the 1st chromosome of rice for tracking；Label D456 is positioned at the dyeing of rice the 4th
Body weighs related QTL:qGW-4LE with grain on the 4th chromosome of rice for tracking；Label D746 is positioned at the dyeing of rice the 7th
Body weighs related QTL:qGW-7LE with grain on the 7th chromosome of rice for tracking；Label D1048 is positioned at the dyeing of rice the 10th
Body weighs related QTL:qGW-10LE with grain on the 10th chromosome of rice for tracking；By mark D140A amplify with raise rice
The identical molecular marker gene type of No. 4 banding patterns is named as A, the molecule mark identical with Lemont banding pattern that D456 will be marked to amplify
Note genotype is named as B, and the molecular marker gene type identical with Lemont banding pattern for marking D746 to amplify is named as C, will
The molecular marker gene type identical with Lemont banding pattern that label D1048 is amplified is named as D；
(4) single plant for whole 4 genotype of A, B, C, D that polymerization procedure (3) describes in the segregating population of selection step (1) description
To obtain the average the smallest rice material of mass of 1000 kernel；Select step (1) description segregating population in polymerization procedure (3) description A,
B, the single plant of any 3 genotype in tetra- genotype of C, D is to obtain average mass of 1000 kernel 4 bases all than polymerization A, B, C, D
Because of the bigger rice material of the single plant of type；Select A, B, C, D of polymerization procedure (3) description in the segregating population of step (1) description
The single plant of any 2 genotype in four genotype is to obtain average mass of 1000 kernel compared in polymerization tetra- genotype of A, B, C, D
The bigger rice material of the single plant of any 3 genotype；It selects to carry step (3) description in the segregating population of step (1) description
Tetra- genotype of A, B, C, D in the single plant of any 1 genotype relatively polymerize tetra- bases of A, B, C, D to obtain average mass of 1000 kernel
Because of the bigger rice material of the single plant of any 2 genotype in type；
(5) by the individual plant selfing mostly generation of the different number of genotype of polymerization procedure (4) description, to obtain the water of different mass of 1000 kernel
Rice homozygosis material.
2. the method according to claim 1 using the medium grain weight type rice of molecular marking supplementary breeding, feature exist
In, in the step (1), Lemont is hybridized with raising rice No. 4, may be selected Lemont as female parent raise rice No. 4 be used as father
This, also can choose and raise rice No. 4 as female parent Lemont and hybridized as male parent.
3. the method according to claim 1 using the medium grain weight type rice of molecular marking supplementary breeding, feature exist
In, in the step (2), using 4 molecular labelings D140A, D456, D746, D1048 come PCR amplification step (1) describe point
Peel off the number of intracorporal single plant, should be actually needed and be determined according to breeding；By taking F2 group as an example, to be obtained in step (4) screening
1 plant of single plant for carrying whole 4 genotype of A, B, C, D, should theoretically detect no less than 256 single plants (calculation method is 4 ×
4 × 4 × 4=256), in another example, to obtain 1 plant of 3 gene carried in tetra- genotype of A, B, C, D in step (4) screening
The single plant of type should theoretically detect no less than 64 single plants (calculation method is 4 × 4 × 4=64)；Example above is theoretically
Supposition, during atual detection, it should the single plant number detected determine according to actual needs；Generally speaking, the list of detection
Strain number mesh is more, then the chance for obtaining the different desired single plants of step (4) description is just bigger.
4. the method according to claim 1 using the medium grain weight type rice of molecular marking supplementary breeding, feature exist
In in the step (3), the banding pattern of molecular marker gene type A is passed through again with using label D140A raising rice No. 4 upper PCR amplifications
The banding pattern that the denaturing polyacrylamide gel electrophoresis of 8% concentration detects is identical, but A is not the genotype of qGW-1YD, due to
D140A and qGW-1YD gene linkage can select the genotype of qGW-1YD indirectly by the genotype of selection A；It similarly can root
It is chain and the genotype of qGW-4LE is selected by indirect selections 1 B gene type according to D456 and qGW-4LE；According to D746 and qGW-
7LE is chain and the genotype of qGW-7LE is selected by indirect selections C genotype；It is chain and logical according to D1048 and qGW-10LE
Indirect selections D genotype is crossed to select the genotype of qGW-10LE.
5. the method according to claim 1 using the medium grain weight type rice of molecular marking supplementary breeding, feature exist
In polymerizeing the average mass of 1000 kernel of the single plant of different number of genotype by sorting from small to large are as follows: polymerization in the step (4)
A, all 4 genotype < polymerize any 2 bases in any 3 genotype < polymerization A, B, C, D in A, B, C, D by B, C, D
Because type < carries any 1 genotype in A, B, C, D；It should be pointed out that above-mentioned sequence size is only applicable to be planted in together
The single plant in the segregating population of same Fn (n >=2 and n is fixed numbers) generation is compared under one environmental condition.
6. the method according to claim 1 using the medium grain weight type rice of molecular marking supplementary breeding, feature exist
In in the step (5), selfing algebra should be actually needed according to breeding and determine.
7. the method according to claim 1 using the medium grain weight type rice of molecular marking supplementary breeding, feature exist
Thousand after the non-groat that, medium grain weight type rice is defined as being planted in after the mature harvest in Hangzhou or Lingshui dries
Rice of the grain dense medium between 21 grams to 26.5 grams.
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| CN104694626A (en) * | 2015-01-22 | 2015-06-10 | 中国水稻研究所 | Method for molecular marker-assisted breeding of middle-short grain rice |
| CN104694625A (en) * | 2015-01-22 | 2015-06-10 | 中国水稻研究所 | Method for breeding long-grain rice under assistance of molecular markers |
| CN105154531A (en) * | 2015-07-28 | 2015-12-16 | 华南农业大学 | Molecular marker for identifying wild type and mutant of 1000-grain weight gene TGW6 of rice |
| CN105441457A (en) * | 2015-12-16 | 2016-03-30 | 中国农业科学院深圳生物育种创新研究院 | Gene qTGW2-1 related with thousand kernel weight of rice under low nitrogen conditions, and molecular marking method thereof |
| CN103421889B (en) * | 2012-10-09 | 2016-06-15 | 华中农业大学 | A kind of method improveing rice grain shape and grain weight |
| CN105985965A (en) * | 2015-02-06 | 2016-10-05 | 中国科学院遗传与发育生物学研究所 | Gene GW7 for controlling grain shape, exterior quality and yield of rice and applications of gene GW7 |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103421889B (en) * | 2012-10-09 | 2016-06-15 | 华中农业大学 | A kind of method improveing rice grain shape and grain weight |
| CN104694626A (en) * | 2015-01-22 | 2015-06-10 | 中国水稻研究所 | Method for molecular marker-assisted breeding of middle-short grain rice |
| CN104694625A (en) * | 2015-01-22 | 2015-06-10 | 中国水稻研究所 | Method for breeding long-grain rice under assistance of molecular markers |
| CN105985965A (en) * | 2015-02-06 | 2016-10-05 | 中国科学院遗传与发育生物学研究所 | Gene GW7 for controlling grain shape, exterior quality and yield of rice and applications of gene GW7 |
| CN105154531A (en) * | 2015-07-28 | 2015-12-16 | 华南农业大学 | Molecular marker for identifying wild type and mutant of 1000-grain weight gene TGW6 of rice |
| CN105441457A (en) * | 2015-12-16 | 2016-03-30 | 中国农业科学院深圳生物育种创新研究院 | Gene qTGW2-1 related with thousand kernel weight of rice under low nitrogen conditions, and molecular marking method thereof |
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