CN116144674B - Rice albino transgenic green leaf mutant gene TWINKLE and application thereof - Google Patents
Rice albino transgenic green leaf mutant gene TWINKLE and application thereof Download PDFInfo
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Abstract
The invention relates to a rice albino-transgenic green leaf mutant gene TWINKLE and application thereof, wherein the CDS sequence of the rice albino-transgenic green leaf mutant gene TWINKLE is shown as SEQ ID No.1, the amino acid sequence of the encoded protein is shown as SEQ ID No.2, the phenotype is albino in seedling stage, and when in 4-5 leaf stage in the field, the leaf starts to turn to be partially green, and the character is continued until the mature stage. Can be used for breeding new rice varieties and identifying the purity of seeds, and provides a new gene and method for breeding pure sterile lines.
Description
Technical Field
The invention relates to the technical field of plant genetic engineering and rice molecular breeding, in particular to a rice albino transgenic green leaf mutant gene TWINKLE and application thereof.
Background
Leaves are main organs for photosynthesis of rice, and have close relation with biological yield and economic yield of rice. Leaf color mutation is a common mutation property of rice, and the mutation types are more. The previous researches on the leaf color mutant materials of the rice are more, and the leaf color mutant mechanism of the rice is discussed through the positioning, cloning and functional researches on the leaf color mutant genes of the rice, and the method has important significance for promoting genetic improvement of the rice, high-yield breeding of the rice and the like.
At present, the following safety problems mainly exist in the production of hybrid rice: firstly, the sterility instability phenomenon caused by the influence of low temperature in the seed production process of the rice photo-thermo-sensitive sterile line affects the purity and the yield of the rice photo-thermo-sensitive sterile line and the two-line hybrid rice. Secondly, in the three-line cross breeding, part of excellent sterile lines are easy to produce fertile pollen during seed production and propagation at a higher temperature, so that a small amount of selfing and maturing are caused, and the purity of the three-line sterile lines and the purity of the hybrid rice are affected.
In breeding practice, rice leaf albino is an ideal marker trait that can be used in breeding. The marker traits currently considered to be applied to hybrid rice production should meet the following conditions: 1. the marking character is obvious, and the identification can be carried out in the seedling stage; 2. the marker character is recessive nuclear gene control; 3. the stable expression of the marker character is not easily influenced by environmental factors; 4. the marker trait expression has no negative effect on the agronomic traits of the sterile line and the hybrid progeny thereof. The leaf color mutant controlled by recessive nuclear genes is utilized, the leaf color mutation characters are led into a sterile line of three lines or two lines by means of hybridization, backcrossing and the like, false seeds can be accurately identified and removed in a seedling stage, and the purity and quality of improved seeds are ensured; the yield per unit area is improved.
Disclosure of Invention
The invention aims to provide a rice albino-transgenic green leaf mutant gene TWINKLE and application thereof, wherein the phenotype of the gene is albino in the seedling stage, the leaves start to turn green partially in the 4-5 leaf stage in the field, the character can be used as a rice breeding mark until the mature stage, false seeds can be accurately identified and removed in the seedling stage, and the purity and quality of improved seeds are further ensured, so that the yield per unit area is improved.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a rice albino-transgenic green leaf mutant gene TWINKLE, the CDS sequence of which is shown as SEQ ID No.1, the amino acid sequence of the coded protein is shown as SEQ ID No.2, the phenotype is albino in seedling stage, and when the leaf stage is 4-5 in the field, the leaf starts to turn green partially, and the character is continued until the mature stage.
The invention also provides application of the albino transgenic green leaf mutant gene TWINKLE in rice breeding.
Furthermore, the application is that the albino transgenic green leaf mutant gene TWINKLE of the rice is used as an early breeding marker, false seed identification and elimination are carried out in the seedling stage, and the purity and quality of improved seeds are ensured.
Furthermore, the application is that the albino green leaf mutation gene TWINKLE of the rice is introduced into a sterile line of three lines or two lines by means of hybridization and backcross, and the albino character of TWINKLE gene is utilized for false seed identification and elimination in the seedling stage, so that the purity and quality of improved seeds are ensured.
Further, the application is that TWINKLE genes are introduced into a rice maintainer line and a sterile line, and three-line and/or two-line sterile lines with albino leaf character in a seedling stage are cultivated; after the hybrid F1 generation obtained by hybrid seed production is identified in the seedling stage, the plants with albino leaves are false hybrids of sterile line selfing, and are removed, so that the seed production risk and loss of three-line and/or two-line hybrid rice can be effectively reduced.
The invention also provides a method for breeding a pure-bred sterile line, which is characterized in that the albino-transgenic green leaf mutant gene TWINKLE of the rice is used as an early breeding marker, false-breeding identification and elimination are carried out in a seedling stage, and the pure-bred sterile line is bred.
Furthermore, the method is to introduce the albino green leaf mutation gene TWINKLE of rice into a three-line or two-line sterile line by means of hybridization and backcross, and to utilize the albino character of TWINKLE gene to perform false seed identification and elimination in the seedling stage so as to select and breed a pure sterile line.
Further, the method comprises the steps of introducing TWINKLE genes into a rice maintainer line and a sterile line, and cultivating a three-line and/or two-line sterile line with albino leaf character in a seedling stage; after the hybrid F1 generation obtained by hybrid seed production is identified in the seedling stage, the plants with albino leaves are false hybrids of sterile line selfing, and are removed, so that the seed production risk and loss of three-line and/or two-line hybrid rice can be effectively reduced.
The invention utilizes a chemical mutagen Ethyl Methylsulfonate (EMS) to mutagenize indica rice western 1B to obtain a genetically stable albino-transgenic green leaf mutant which is named twinkle. Wild Type (WT) plants exhibit green leaves throughout the growth phase, except for the mature phase, and compared to wild type, mutant twinkle leaves whiten during the seedling phase and begin to turn green partially during the 4-5 leaf phase in the field, the trait persisting until the mature phase. Through genetic analysis and gene location, it is determined that rice albino transgenic green leaf trait TWINKLE is recessive gene control, single base substitution occurs to 13 th exon of LOC_Os06g45980 gene, so that coded amino acid Gly is changed into Asp, CDS sequence is shown as SEQ ID No.1, and amino acid sequence of coded protein is shown as SEQ ID No. 2. Further amplified genomic fragments (comprising TWINKLE upstream of promoter, coding sequence, downstream of terminator) from wild type gDNA were transformed into twinkle mutants for complementation verification, and in transgenic plants, the mutant phenotype was completely restored. Further, it was confirmed that albino green leaf mutation of rice was controlled by the mutant gene TWINKLE. Therefore, the albino green leaf mutant gene TWINKLE of rice can be used for rice breeding.
The invention has the beneficial effects that: the invention discloses a rice albino-transgenic green leaf mutant gene TWINKLE, the CDS sequence of which is shown as SEQ ID No.1, the amino acid sequence of the coded protein is shown as SEQ ID No.2, the phenotype is albino in seedling stage, and when in 4-5 leaf stage in field, the leaf starts to turn green partially, and the character is continued until the mature stage. The gene can be used for breeding new varieties of rice and identifying the purity of seeds, and provides a new gene and a new method for breeding pure sterile lines.
Drawings
FIG. 1 is a diagram of the results of a positioning analysis and complementary verification of TWINKLE;
a: TWINKLE locating the analysis result;
B-D: sequencing a complementary positive transgenic plant;
E-G: phenotypic observations of wild type (E), mutant twinkle (F), com-twinkle plants (G).
FIG. 2 shows the results of pigment content analysis.
FIG. 3 shows the results of related gene expression analysis;
TWINKLE expression of the gene;
And B, C, chloroplast development, photosynthesis and pigment metabolism related gene expression.
Detailed Description
The technical scheme of the invention will be further described in detail below with reference to specific embodiments. It is to be understood that the following examples are illustrative only and are not to be construed as limiting the scope of the invention. All techniques implemented based on the above description of the invention are intended to be included within the scope of the invention. It should be noted that, the experimental materials in the examples of the present invention are all commercially available, and the experimental methods without specific conditions are usually performed according to conventional conditions or according to the recommended conditions of the experimental material manufacturer.
EXAMPLE 1 obtaining and morphological observations of albino-transgenic Green leaf mutants of Rice
A genetically stable albino transgenic green leaf mutant named twinkle is obtained by mutagenesis of indica rice western 1B with a chemical mutagen Ethyl Methylsulfonate (EMS). Wild Type (WT) plants exhibit green leaves throughout the growth phase, except for the mature phase, and compared to wild type, mutant twinkle leaves whiten during the seedling phase and begin to turn green partially during the 4-5 leaf phase in the field, the trait persisting until the mature phase.
Example 2TWINKLE Gene mapping
The mutant twinkle is taken as a female parent, Jin is recovered 10 as a male parent, F1 generation is obtained by hybridization, F2 generation is obtained by F1 generation selfing, the character of F2 generation is separated, and chi-square verifies that the normal phenotype and the mutant phenotype accord with the separation ratio of 3:1, which indicates that the mutant character is controlled by a pair of recessive nuclear genes and is named as TWINKLE.
Gene localization was performed using mutant plants in F2 generation, and TWINKLE gene was finally located between chromosome 6 primer markers Chr6-25.40 and Chr6-27.90, with a physical distance of 2.50M (FIG. 1A).
The wild type and mutant twinkle leaf samples are subjected to whole genome sequencing, and are compared by software IGV_2.13.2, and finally single base substitution (G is changed into A) of 13 th exon of LOC_Os06G45980 gene is found in the interval between Chr6-25.40 and Chr6-27.90, so that coded amino acid Gly is changed into Asp (figure 1A), and the TWINKLE gene is primarily proved to be a mutant gene of LOC_Os06G45980, wherein the CDS sequence is shown as SEQ ID No.1, and the amino acid sequence of coded protein is shown as SEQ ID No. 2.
EXAMPLE 3twinkle mutant complementation verification
Amplifying genome fragment (comprising TWINKLE upstream of promoter, coding sequence and downstream of terminator) from wild gDNA, recombining the fragment into pCAMBIA1301 vector, extracting plasmid after positive cloning, transforming twinkle mutant with recombinant vector and pCAMBIA1301 empty vector by agrobacterium transformation method to obtain Com-twinkle and Ve-twinkle positive transgenic plant respectively.
After obtaining transgenic plants, carrying out GUS staining preliminary identification on the transgenic plants by using a GUS reporter gene on a pCAMBIA1301 vector, then extracting DNA of the preliminary identified transgenic plants for amplification and sequencing analysis, and identifying positive transgenic plants Com-twinkle (figure 1) by comparing sequencing results: ① Determining whether the wild-type gene is transferred, and performing reverse sequencing due to poor forward sequencing effect (FIG. 1B); ② Determining whether the wild-type vector is ligated (FIG. 1C); ③ It was determined whether the infected mutant calli were reverse sequenced (FIG. 1D).
Phenotype observation of the positive transgenic plants revealed that Com-twinkle plants restored the overall plant height and leaf phenotype (fig. 1E-G) compared with the mutant phenotype, further demonstrating that TWINKLE gene is a mutant gene of loc_os06G45980 gene.
Example 4 functional analysis
1. Pigment content and photosynthesis analysis
Pigment content measurement is carried out on wild type and mutant leaves at the tillering stage, and the result shows that compared with the wild type, the chlorophyll a, chlorophyll b, carotenoid and total chlorophyll content in the green and white leaves of the mutant are extremely reduced (figure 2); meanwhile, the photosynthetic data analysis shows that compared with a wild type, the net photosynthetic rate Pn and the transpiration rate Tr of the green leaf of the mutant are obviously reduced, and the net photosynthetic rate Pn, the stomatal conductance Gs, the intercellular CO 2 concentration Ci and the transpiration rate Tr of the white leaf of the mutant are extremely obviously reduced (table 1). It is shown that the mutation of TWINKLE gene affects the pigment content of plant leaf and photosynthesis.
TABLE 1 photosynthesis analysis results
2. Expression analysis
Extracting wild-type root, stem, leaf, sheath and spike RNA, reversely transcribing into cDNA, analyzing TWINKLE gene expression condition, finding that the gene is expressed in root, stem, leaf, sheath and spike, belonging to constitutive expression (figure 3A); meanwhile, the analysis of the expression of some chloroplast development-related, photosynthesis-related and pigment metabolism-related genes was carried out, and these genes were affected to different extents in the leaves of the mutants as compared with the wild type (FIGS. 3B, C).
The foregoing description of the preferred embodiments of the invention is not intended to be limiting, but rather to cover all modifications, equivalents, alternatives, and improvements that fall within the spirit and scope of the invention.
Claims (7)
1. The CDS sequence of the rice albino transgenic green leaf mutant gene TWINKLE is shown as SEQ ID No.1, the amino acid sequence of the coded protein is shown as SEQ ID No.2, the phenotype is albino in the seedling stage, and when the leaf stage is 4-5 in the field, the leaf starts to turn green partially, and the character is continued until the mature stage.
2. The application of the rice albino green leaf mutant gene TWINKLE in rice breeding according to claim 1, wherein the application is that the rice albino green leaf mutant gene TWINKLE is used as an early albino character breeding marker, and false seed identification and elimination are carried out in a seedling stage.
3. The use according to claim 2, wherein the use is to introduce the albino green leaf-transformed rice mutant gene TWINKLE into a sterile line of three lines or two lines by means of hybridization and backcrossing, and to perform the identification and elimination of the false seeds in the seedling stage by using the albino trait of the mutant gene TWINKLE.
4. The use according to claim 3, wherein the use is to introduce a mutant gene TWINKLE into a rice maintainer line and a sterile line, and to cultivate a three-line and/or two-line sterile line with a seedling albino leaf trait; after the hybrid F1 generation obtained by hybrid seed production is identified in the seedling stage, the plants with albino leaves are false hybrids of sterile line selfing, and are removed, so that the seed production risk and loss of three-line and/or two-line hybrid rice can be effectively reduced.
5. A method for breeding a pure sterile line is characterized in that the rice albino green leaf-transformed mutant gene TWINKLE of claim 1 is used as an early albino character breeding marker, and false seed identification and elimination are carried out in a seedling stage to breed pure sterile line.
6. The method for breeding a pure sterile line according to claim 5, wherein the method is characterized in that the albino transgenic green leaf mutant gene TWINKLE of the rice in claim 1 is introduced into a sterile line of three lines or two lines by means of hybridization and backcross, and the albino character of the mutant gene TWINKLE is utilized for false seed identification and elimination in a seedling stage to breed the pure sterile line.
7. The method for breeding a pure sterile line according to claim 6, wherein the method is to introduce a mutant gene TWINKLE into a rice maintainer line and a sterile line and cultivate a three-line and/or two-line sterile line with seedling albino leaf character; after the hybrid F1 generation obtained by hybrid seed production is identified in the seedling stage, the plants with albino leaves are false hybrids of sterile line selfing, and are removed, so that the seed production risk and loss of three-line and/or two-line hybrid rice can be effectively reduced.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1402968A (en) * | 2002-10-21 | 2003-03-19 | 浙江大学 | Selective breeding process of hybrid rice with greening leaf color label carried in its seedling stage |
| CN106086036A (en) * | 2016-07-14 | 2016-11-09 | 武汉大学 | Rice seedling blade albefaction character gene Oscaac1 and application thereof |
| CN108913668A (en) * | 2018-07-27 | 2018-11-30 | 西南大学 | Rice albefaction turns albumen and the application of greenery gene VAL1 and its coding |
| CN110283925A (en) * | 2019-05-10 | 2019-09-27 | 武汉双绿源创芯科技研究院有限公司 | A method of for identifying rice green genetic fingerprints |
| WO2022115902A1 (en) * | 2020-12-01 | 2022-06-09 | Commonwealth Scientific And Industrial Research Organisation | Cereal grain with thickened aleurone |
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| US20110131679A2 (en) * | 2000-04-19 | 2011-06-02 | Thomas La Rosa | Rice Nucleic Acid Molecules and Other Molecules Associated with Plants and Uses Thereof for Plant Improvement |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1402968A (en) * | 2002-10-21 | 2003-03-19 | 浙江大学 | Selective breeding process of hybrid rice with greening leaf color label carried in its seedling stage |
| CN106086036A (en) * | 2016-07-14 | 2016-11-09 | 武汉大学 | Rice seedling blade albefaction character gene Oscaac1 and application thereof |
| CN108913668A (en) * | 2018-07-27 | 2018-11-30 | 西南大学 | Rice albefaction turns albumen and the application of greenery gene VAL1 and its coding |
| CN110283925A (en) * | 2019-05-10 | 2019-09-27 | 武汉双绿源创芯科技研究院有限公司 | A method of for identifying rice green genetic fingerprints |
| WO2022115902A1 (en) * | 2020-12-01 | 2022-06-09 | Commonwealth Scientific And Industrial Research Organisation | Cereal grain with thickened aleurone |
Non-Patent Citations (9)
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