CN114438101A - Allele with transparent rice appearance and low amylose content and application thereof - Google Patents

Abstract

The invention discloses an allele with transparent rice appearance and low amylose contentDu1 ^ΔA121 Belonging to the field of biotechnology. The gene is in riceDu1The 361 st to 363 th nucleotides of the 1 st exon of the gene coding region are deleted, and the gene coding region contains a gene sequence shown as SEQ ID No.1 and a coding region sequence shown as SEQ ID No. 2. The invention also discloses aDu1 ^ΔA121 The encoded protein comprises an amino acid sequence shown as SEQ ID No. 3. The amylose content control gene can also be applied to breeding of new rice varieties with transparent rice appearance and low amylose content. The present invention is portableDu1 ^ΔA121 Mutant of genetas1The AC is about 11.0%, which is reduced by about 4.5%, compared with the soft rice control Nanjing 9108 (the AC is 8.7%), the kernel transparency is obviously better.

Description

Allele with transparent rice appearance and low amylose content and application thereof

Technical Field

The invention relates to an allele Du1 with transparent rice appearance and low amylose content^ΔA121And the application thereof, belonging to the field of molecular genetics.

Background

Rice is an important grain crop in the world, and plays a significant role in guaranteeing grain safety. In recent 30 years, the rice yield of China is continuously improved through the implementation of plans such as high-yield breeding, ultrahigh-yield breeding, super rice breeding and green super rice breeding, but the problem of rice quality is more and more prominent. In recent years, with the improvement of the living standard of people in China, the main contradiction between rice production and consumption is changed from the shortage of total amount to the surplus of structure. Under the new trend, the cultivation of new high-quality rice varieties and the development of high-quality rice industries, and the meeting of the increasing demands of people on good life become important tasks for serving three farmers. The development of high-quality rice research meets the current market requirements.

The relationship between the taste quality of rice and the starch content is tight. There are two main types of starch that constitutes rice, namely amylose and amylopectin, among which amylose is considered as the most important determinant of the quality of cooked taste of rice (Zhu Da Wei et al, 2015). The rice Wx gene encodes granular starch synthase (GBSS), which is a major gene controlling amylose synthesis and directly affects the amylose content in rice endosperm and pollen. In addition, the Du1(dull endosperm 1) gene encodes a pre-mRNA splicing factor which is directly involved in regulating the splicing efficiency of pre-mRNA of the starch synthesis related gene in a complex form to influence the transcription quantity of mature mRNA. To date, 8 loci have been found to cause dark endosperm mutations, such as du-1, du-2, du-3, du-4, du5, du2035, du (2120), du (EM47) (Isshiki et al 2000). Studies have shown that the dark endosperm mutation only reduces AC and has no effect on amylopectin properties, e.g. the length of the amylopectin is unchanged. Studies have shown that du-1 and du-2 do not form normally spliced protein factors, which makes the splicing process of Wxb pre-mRNA abnormal, and thus reduces the amylose content of the mutants (Isshiki et al, 2000).

A novel rice variety with lower Amylose Content (AC) (7-12 percent) is commonly called soft rice, and the rice of the novel rice variety has the advantages of softness without rotting, sweetness, dainty taste, good swelling property, high elasticity, difficult hardening after cooling, small retrogradation degree and the like, and is the typical characteristic of high-quality rice in most areas of China. In recent years, low-AC japonica rice varieties such as Yunjun 29, Longjing 38, Nanjing 46, Shanghai 1212 and the like in Yunnan, Heilongjiang, Jiangsu and Shanghai have outstanding taste and quality, and are popular with consumers and widely planted. The consumption group of low AC japonica rice is expanding year by year, and the market prospect of related products is becoming wide.

Although the low AC japonica rice is popular and touted in the market in taste, the two defects are not well solved for a long time: 1. most of the existing low AC varieties have poor appearance quality. At present, most of japonica rice varieties with low AC (about 9 percent) in production are cloudy and poor in transparency, are commonly called semi-glutinous rice or stiff rice, reduce the commodity value of the rice, and are difficult to compete with rice in northeast China and Japan (transparent and well-sold) in appearance quality. The transparency of rice can be improved by increasing the water content of the rice to more than 17%, but the rice is often not storable and even mildewed. When the AC is 10-13%, the rice can keep good taste quality, and the appearance quality of the rice is obviously improved and is transparent. However, such germplasm and gene resources are lacking in current rice breeding. 2. The available rice is transparent in appearance, the germplasm of the available rice is lack of low AC, and the available genotype is single. In recent 20 years, the low AC variety bred in Yangtze triangle area mainly utilizes the allelic variation gene Wx-mp of Guandong 194 of Japanese soft rice variety, and the AC content is about 9%. In recent years, no new soft rice gene resource has been discovered, and low AC germplasm with excellent appearance quality and high breeding utilization value is lacking, so that market demand is difficult to meet. Therefore, by digging and utilizing regulatory genes and materials of soft rice with excellent appearance quality, the change of the current situation that the appearance of the existing low AC does not reach the standard is urgent need for high-quality breeding.

There is no report of allelic variation of Du1 gene.

Disclosure of Invention

The technical problem is as follows: the technical problem to be solved by the invention is to provide a novel allele Du1 related to the transparent appearance of rice and low amylose content^ΔA12。

The technical problem to be solved by the invention is to provide the allele Dul^ΔA12The encoded protein.

The technical problem to be solved by the invention is to provide the application of the allele in controlling the apparent transparency and/or low amylose content of rice.

The technical problem to be solved in the end of the present invention is to provide a method for obtaining or identifying rice with low amylose content which is transparent in appearance.

The technical scheme is as follows: in order to solve the above technical problems, the present invention provides an allele Dul^ΔA12The allele Du1^ΔA12The gene related to the low amylose content and transparent rice appearance is a gene related to the low amylose content of rice, the gene has the accession number of XM-015758339.2, and the 361 st to 363 th nucleotides of the 1 st exon of the coding region of the rice Du1 gene are deleted, and the gene comprises a gene sequence shown as SEQ ID No.1 and a coding region sequence shown as SEQ ID No. 2.

Specifically, the invention utilizes Jiangsu province rice variety Su reclamation 118 as background material to carry out ionizing radiation to obtain more than ten thousand parts of M₀Mutant, selfing and breeding. And identifying and screening by taking the grain with transparent appearance and 8% -12% of AC as target characters. A mutant which has AC of 11.0% and transparent rice appearance is obtained by screening under the condition that the water content of rice is 12%, and is named as tas1 (transparent-Appearance and soft 1), the mutant tas1 is preserved in China general microbiological culture Collection center (CGMCC) at 22 months at 2022, with the preservation number of CGMCC No.24030 and the classified name of rice (Oryza sativa) with the preservation address of No.2 Hosier No.1 of the sunward area of Beijing city.

Specifically, the Du1 gene of the mutant is found to have non-frameshift weak mutation through research, the mutant tas1 has deletion of nucleotides 361 to 363 of the 1 st exon of the coding region of the rice Du1 gene with the gene accession number of XM _015758339.2 (figure 5), and the sequence is shown as SEQ ID NO. 1. Compared with the wild type, the mutation causes the deletion of alanine at position 121 in the protein coded by the mutant gene (figure 5), the sequence is shown as SEQ ID NO.3, and other amino acids have no mutation. Therefore, Du1 gene of mutant tas1 is a new allelic variation occurring on the basis of wild type Du1 and is named as Du1^ΔA121。

The present invention also includes the allele Du1^ΔA12The amino acid sequence of the encoded protein is shown as SEQ ID No. 3.

The present disclosure also includes expression cassettes, recombinant vectors or cells containing the alleles.

The present invention also includes the allele Du1^ΔA12The protein, the expression cassette, the recombinant vector or the cell are applied to rice cross breeding and variety improvement.

Wherein, the application comprises the application of controlling the appearance transparency of the rice and/or the low amylose content.

The present invention also includes a method for obtaining rice with a low amylose content that is transparent in appearance, comprising the steps of:

1) the rice contains the allele Du1^ΔA12(ii) a Or

2) The rice expresses the allele Du1^ΔA12The encoded protein.

Wherein the method comprises the steps of transgenosis, hybridization, backcrossing or asexual propagation.

The present invention also includes a method for identifying rice plants comprising said allele Du1^ΔA12The rice expressing the protein or the rice obtained by the method comprises the following steps:

1) identifying whether said rice comprises said allele; or the like, or, alternatively,

2) identifying whether said rice expresses said protein.

Has the advantages that: compared with the prior art, the invention has the following advantages:

1) the invention obtains a low AC related protein with transparent rice appearance and a gene Du1 for coding the protein^ΔA121. The gene encodes a pre-mRNA splicing factor Du1^ΔA121By influencing the splicing efficiency of the precursor mRNA of the AC major regulatory gene Wx, the expression of Wx is reduced and AC is reduced. Compared with wild plants, the wild plants carry homozygous Du1 by about 15.5 percent of AC^ΔA121The mutant tas1 of the gene has the AC of about 11.0 percent, is reduced by about 4.5 percent, and has obviously better kernel transparency compared with the soft rice control Nanjing 9108 (the AC is 8.7 percent). Cloning a new low AC regulation gene with transparent rice appearance by using a new low AC resource with transparent rice appearanceThe rice appearance transparent low AC regulation gene library and the analysis of the corresponding friendship phenotype formation mechanism have important theoretical and practical significance. The AC has proper reduction value, obvious trend and obvious effect, and can effectively improve the palatability and the taste quality.

2) The rice disintegration value (BDV) of the rice mutant tas1 material obtained by the invention is 1321.7 +/-87.7 cP, and compared with the wild type 989.7 +/-111.8 cP, the BDV is increased by a proper value and has obvious difference; the rice recovery value (CSV) of the rice mutant tas1 material discovered by the invention is 777.0 +/-16.8 cP, and compared with 1112.3 +/-21.9 cP of a wild type, the CSV has proper reduction value and obvious difference; the rice peak time (PeT) of the rice mutant tas1 material discovered by the invention is 5.7 +/-0.1 min, and compared with the wild type 5.9 +/-0.1 min, the PeT is reduced by a proper value and has obvious difference. The RVA spectrum viscosity characteristics of the rice represented by the 3 indexes in the mutant are obviously superior to those of a wild type control, and the mutant can be used for cultivating cooking taste quality improvement plants.

3) The invention utilizes F obtained by hybridization of the mutant tas1 and the conventional japonica rice variety Nanjing 51₂In the population, 24 single plants are randomly selected for rice transparency observation, AC detection and Du1 gene sequencing, and the result shows that the rice transparency is better in all the plants, and all the plants with the AC of more than 14 percent are Du1Du1 or Du1Du1^ΔA121Genotype, Du1 for all plants with AC less than 14%^ΔA121Du1^ΔA121Genotype, Du1^ΔA121The gene was completely co-segregating with a phenotype of less than 14% of AC. The gene is transferred into other conventional rice varieties by conventional technical means such as hybridization, backcross and the like, and a new low AC material with transparent rice appearance can be cultivated.

Drawings

FIG. 1 phenotypic analysis; a: amylose Content (AC) assay; b: observing the appearance transparency of polished rice; SK188 is Suzhou reclamation 118, tas1 is mutant tasl, NG9108 is Nanjing 9108;

FIG. 2 shows the overall variation of the RVA characteristic values; NG46 is Nanjing 46;

FIG. 3 genetic analysis;

FIG. 4 alignment of coding region sequences of Wx genes; SK188 is Suzhou reclamation 118, tas1 is mutant tas 1;

FIG. 5 Du1 gene structure and Du1^ΔA121Schematic representation of allelic sequence differences;

FIG. 6 Wx Gene expression analysis.

Detailed Description

Embodiments of the present invention will be described in detail below with reference to examples, but those skilled in the art will appreciate that the following examples are only illustrative of the present invention and should not be construed as limiting the scope of the present invention. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products commercially available.

Example 1: acquisition and phenotypic characterization of mutant tas1

1. Obtaining mutant tas1

The background material selected by the invention is the reclamation of Suzhou 118 (purchased from Jiangsu reclamation seed industry Co., Ltd.), the variety is a new late-maturing Zhongjing variety bred by the food crop research institute of the agricultural academy of sciences of Jiangsu province, has about 155 days of the whole growth period, is suitable for planting in Suzhou province and Ningzhengyangyang hilly areas of Jiangsu province, has excellent comprehensive agronomic characters, is popularized and applied in large area in production, and is deeply popular in the market. The Su-cultivated 118 plants are compact in type, strong in tillering force, light green in leaves, good in group uniformity, good in lodging resistance, good in color change in the mature period, about 15.5% of amylose content, and transparent in rice appearance. The mutant material with transparent appearance and properly reduced AC of the rice is created by using the variety through a physical-chemical mutagenesis technology, is used as a research object, is analyzed from a biochemical and molecular level, and is finally applied to new variety breeding, and the strategy is a feasible way for obtaining a low AC product with good appearance quality of the rice.

The invention utilizes Jiangsu province rice variety Su reclamation 118 as background material to carry out ionizing radiation, and more than ten thousand M is obtained₀Mutant, selfing and breeding. And identifying and screening by taking the grain with transparent appearance and 8% -12% of AC as target characters. A mutant, designated as tas1 (transparent-apple), was selected in which AC was 11.0% and rice appearance was clear when a portion of rice with a water content of 12% was obtainedarance and soff 1), the biological material is preserved in China general microbiological culture collection center (CGMCC) at 22 months and 2 months in 2022, the preservation number is CGMCC No.24030, the biological material is classified and named as rice (Oryza sativa), and the preservation address is Beijing city Shangyang Wen Luo No.1 Hospital No.2 of the sunward district.

2. AC determination and rice appearance observation

The mutant tas1 obtained from wild type variety Su Ke 118 (conventional japonica rice) and low AC control variety Nanjing 9108 (Wx)^mpGenotype) was determined with reference to the standard issued by the ministry of agriculture NY147-88, 4 reference standard samples (AC: 1.5%, 10.6%, 16.4% and 25.6%) were purchased from the rice institute of china. The results are shown in figure 1A with an AC of threo 118 of 15.5% ± 0.1 and an AC of mutant tas1 of 11.0% ± 0.2, significantly lower than its wild type, but still higher than southern japonica 9108(AC of 8.7% ± 0.1).

The transparency is one of the main indexes for measuring the appearance quality of rice. The invention uses wild type soda 118 (transparent) and Nanjing 9108 (in cloud form) polished rice as a control to observe the appearance of the polished rice of the mutant. Rice is hulled by a huller (SY88-TH, Korean Bilong) to obtain coarse rice, refined by a small-sized refined rice machine (BLH-3120, Burley constant, Taizhou) to obtain refined rice, and water content of the refined rice is measured by a water analyzer (Mettler, Switzerland) to ensure consistent water content of the test sample. As shown in FIG. 1B, the rice of Su Ken 118 was transparent at a water content of 12%, but Nanjing 9108 was cloudy and poor in transparency, and mutant tas1 was transparent as in Su Ken 118. The mutant is shown to have a significantly reduced AC compared with the wild type, but still can keep better transparency in appearance.

Thus, tas1 can be considered a whole new piece of rice appearance transparent low AC material.

3. Rice RVA spectrum viscosity determination

The Cooking and Cooking Quality (ECQ) is a direct factor influencing the selection of consumers, and is the most important evaluation index in the rice Quality constitution. Although China has gone out of the national standard of sensory evaluation method for rice cooking edible quality (GB/T15682-2008), the rice quality cannot be accurately identified by a manual tasting mode because the influence of subjective factors cannot be completely eliminated. There is a close relationship between the RVA profile characteristic of rice and the quality of cooked flavour of rice, as measured using a rapid viscosity analyzer (RVA Super 4, NEWPORT SCIENTIFIC, Australia) with parameter settings made in accordance with the American Association of cereal chemists AACC61-01 and 61-02 protocols. Previous studies have shown that rice with good taste quality tends to have a large disintegration value (BDV), rice with relatively non-retrogradation in cold rice tends to have a small recovery value (CSV), and in addition, the peak time PeT refers to the time taken for the sample to reach the peak viscosity, and generally the smaller the PeT, the better the swelling and breakage of the starch grains. Results as shown in table 1, BDV of mutant tas1 was 1321.7 ± 87.7cP, significantly greater than 989.7 ± 111.8cP of wild type; the CSV of mutant tas1 is 777.0 + -16.8 cP, which is significantly less than the 1112.3 + -21.9 cP of wild type; PeT of the mutant tas1 is 5.7 +/-0.1 min, which is obviously less than 5.9 +/-0.1 min of the wild type. This data shows that mutant tas1 has better taste quality compared to wild type, cold rice is relatively non-retrograded, and starch granules have better expansion and breakage during cooking. Overall variation of RVA characteristics as shown in figure 2, the RVA profile of mutant tas1 differs from the wild type, which shows terminal upwarp due to FV greater than PV, but the mutant tended to low AC control variety nanjing 46, i.e. FV less than PV, showing no terminal upwarp.

TABLE 1 RVA spectral eigenvalues

Example 2: genetic analysis and cloning of target genes

1. Genetic analysis

Using Su-zao 118 as female parent and mutant tas1 as male parent to prepare hybrid combination to obtain F₂The population (n 140) was subjected to genetic analysis. As shown in FIG. 3, low AC individuals (AC < 14%): the separation ratio of high AC single plant (AC > 14%) tends to 1: 3 (chi)²＝1.61＜χ² _0.05，1) It was shown that the low AC phenotype in mutant tas1 was controlled by a stealth monogene.

2. Cloning of genes

The Wx gene is a main effective gene for regulating and controlling the starch content of rice, and is firstly identified by utilizing a target gene resequencing technology. Extracting genome DNA of leaf blades of the Suzhou reclamation 118 and the mutant tas1 respectively by using a CTAB method, and respectively carrying out PCR amplification by using the extracted genome DNA as a template. The upstream primer used for amplification is 5'-cggtgcccaacagaaaccaca-3', and the downstream primer is 5'-cacccagaagagtacaacat-3'. The PCR system was a DNA template (20 ng. mu.L)^-1)2μL，10×PCR buffer 2μL，MgCl₂(5mmol L^-1)2μL，dNTP(2mmol L^-1)2 μ L, forward primer (2 μmol L)^-1)2 μ L, downstream primer (2 μmol L)^-1)2 μ L, Taq DNA polymerase (5U μ L)^-1)0.2μL，ddH₂O7.8. mu.L. The amplification condition is 94 ℃ for 5 min; 30s at 94 ℃, 30s at 55 ℃, 4min at 72 ℃ and 35 cycles; extension was carried out at 72 ℃ for 10min to terminate the reaction. PCR was performed in an Eppendorf Mastercycle thermocycler. Separating the amplified product by agarose gel electrophoresis, cutting gel, recovering target band, and sequencing by Nanjing Ongzhike Biotechnology Co. Alignment of the Wx gene of Nipponbare, whose gene accession number is EU770319 in Suzhou 118, mutant tas1 and databases was carried out using BioXM2.6 software. As shown in FIG. 4, Wx sequence in Sunao 118 is identical to Nipponbare and is Wx^bAllelic type, no variation in the Wx gene in mutant tas1 compared to wild type thresh 118, indicates that the mutant phenotype is caused by other genetic variations.

Secondly, a target gene re-sequencing technology is utilized to identify the reported Du1 gene, the upstream primer used for amplification is 5'-CGACTAATCACAAGCGTCTT-3', and the downstream primer is 5'-CCATCCCAGTTCACTACCC-3'. The PCR system was as described in the previous paragraph. The amplification condition is 94 ℃ for 5 min; 30s at 94 ℃, 30s at 55 ℃, 5min at 72 ℃ and 35 cycles; extension was carried out at 72 ℃ for 10min to terminate the reaction. PCR was performed in an Eppendorf Mastercycler thermocycler. Gel cuts were recovered and sequenced as described in the previous paragraph. Alignment of the rice Du1 gene with sovereign 118, mutant tas1 and database under the accession number XM _015758339.2 was performed using the bioxm2.6 software. The results show that the Du1 sequence in Suzhou 118 is identical to Nipponbare, as shown in FIG. 5, Du1 gene in mutant tas1 is compared with that in NipponbareWild type thresh 118, which has a deletion of nucleotides 361 to 363 of exon 1 of the coding region, the mutation resulting in the deletion of alanine 121 in the encoded protein, and no mutation of other amino acids. No report of this allelic variation of Du1 gene exists at present. Therefore, Du1 gene of mutant tas1 is a new allelic variation occurring on the basis of wild type Du1 and is named as Du1^ΔA121。

Example 3 Co-separation and preliminary functional analysis

1. Coseparation analysis

To verify that the AC reduction phenotype in mutant tas1 was due to the Du1 gene mutation, cosegregation analysis was performed genetically. Utilizing F in example 2₂Individual plants of the population (n 140) were subjected to AC assay and genotype determination. The AC determination is described in example 1. The genotype identification is carried out by a method of directly sequencing a PCR product, wherein an upstream primer used for amplification is 5'-CCTCCTCCCTGCTACTCCAC-3', and a downstream primer is 5'-TAGTCCCCAATTTCAGGTATGCTT-3'. The PCR system is shown in example 2. The amplification condition is 94 ℃ for 5 min; 30s at 94 ℃, 30s at 62 ℃, 50s at 72 ℃ and 34 cycles; extension was carried out at 72 ℃ for 3min to terminate the reaction. PCR was performed in an Eppendorf Mastercycler thermal cycler. The gel cut recovery and sequencing method is described in example 2.

As shown in Table 2, in the above-mentioned F₂38 individuals of the population (n 140) identified wild-type genotypes, all exhibiting high AC (AC > 14%); obtaining 74 heterozygous individuals, wherein all the individuals show high AC (AC is more than 14%); obtaining homozygous Du1^ΔA12128 individuals of the mutant genotype, all showed low AC (AC < 14%), indicating Du1^ΔA121And low AC phenotype, namely, the AC reduction phenotype in the mutant tas1 is proved to be caused by Du1 gene mutation. The specific individual AC assay and gene identification are shown in Table 3.

TABLE 2 coseparation analysis

TABLE 3 Individual AC assay and Gene identification

WT represents a wild type; he represents a heterozygote type; ho stands for homozygous mutant

2. Wx Gene expression level detection

Previous studies showed that the Du1 gene encodes a pre-mRNA splicing factor that can decrease Wx expression and AC by affecting the splicing efficiency of pre-mRNA of the AC major regulatory gene Wx. In the present invention, the expression of the Wx gene in wild type threo reclamation 118 and mutant tas1 were further compared. Endosperm RNAs of threo reclamation 118 and mutant tas1 at 6 days, 9 days and 12 days after flowering were extracted and reverse-transcribed into cDNAs, and the obtained cDNAs were used as templates for fluorescent quantitative PCR detection. OsActin-1 is used as an internal reference gene (an upstream primer is 5'-CCAAGGCCAATCGTGAGAAGA-3', a downstream primer is 5'-AATCAGTGAGATCACGCCCAG-3'), and the expression level of the Wx gene (an upstream primer is 5'-ATTCCTTCAGTTCTTTGTCTATCTCA-3', and a downstream primer is 5'-ATGGTGGTTGTCTAGCTGTTGC-3') is identified by a fluorescent quantitative PCR method. The amplification system was 5. mu.L of cDNA template (1ng of RNA obtained by reverse transcription and diluted 3-fold), 10. mu.L of 2 XChamQ SYBR qPCR Master Mix (Novozan Biotech Co., Ltd.), and upstream primer (10. mu. mol L)^-1) 0.4. mu.L, downstream primer (10. mu. mol L)^-1)0.4μL，ddH₂O4.2. mu.L. The reaction condition is 95 ℃ for 30 s; 10s at 95 ℃, 30s at 60 ℃ and 40 cycles; the reaction was terminated at 95 ℃ for 15s, 60 ℃ for 60s, and 95 ℃ for 15 s. PCR was performed in a fluorescent quantitative PCR instrument (Roche Applied Science LightCycler 480), and data were derived and analyzed using the self-contained derivation function. The results are shown in fig. 6, where the expression of Wx gene in mutant tas1 was reduced in endosperm at 6, 9 and 12 days post anthesis compared to wild type threw 118, only 58.4%, 60.3% and 64.8% of wild type respectively. The above results indicate that the deletion of the 361 st to 363 th nucleotides of the Du1 gene in the mutant results in the reduction of the Wx gene expression level in endosperm during the filling stageAnd ultimately a reduction in AC.

Example 4 Breeding applications

In order to utilize the novel low-AC material tas1 with transparent rice appearance in breeding, the mutant tas1 is hybridized with the conventional japonica rice variety Nanjing 51 (carrying the wild Du1 gene and having transparent appearance and AC of 16.0% + -0.2). In the obtained F₂In the population, 24 single plants are randomly selected for rice transparency observation, AC detection and Du1 gene sequencing, and the result shows that the rice transparency of all detected plants is better; furthermore, as shown in Table 4, all plants showing high AC (> 14%) were wild-type (Du1Du1) or heterozygous (Du1Du1)^ΔA121) Genotype, all plants showing low AC (AC < 14%) were homozygous mutant genotypes (Du 1)^ΔA121Du1^ΔA121)，Du1^ΔA121The gene was completely cosegregated with a phenotype of less than 14% of AC, indicating that Du1 was carried by the genotype screen^ΔA121Du1^ΔA121The selection of low AC phenotype in breeding early generation materials can be realized by using a single plant of the genotype. The specific individual AC assay and gene identification are shown in Table 5. The results further show that the gene can be transferred into other conventional rice varieties by conventional technical means such as hybridization, backcross and the like, and a novel low-AC material with transparent rice appearance can be cultivated.

TABLE 4 genotype and AC detection analysis

TABLE 5 Individual plant AC assay and Gene identification

WT represents a wild type; he represents a heterozygote type; ho stands for homozygous mutant

Sequence listing

<110> agricultural science and academy of Jiangsu province

<120> allele with low amylose content and transparent rice appearance and application thereof

<160> 3

<170> SIPOSequenceListing 1.0

<210> 1

<211> 4592

<212> DNA

<213> tas1 mutant (transparent-apearance and soft 1)

<400> 1

attcttcctc acgacctcaa aaacccaaac caatctactc cgccgccgcc gccgcgatgg 60

tgttcgtccg cgcgccggac gggaggaccc accacgtcga cctcgacccc tccaccgcca 120

cgctcgccga cctcacggcc tccgcctccc gcgtctgcgg cggcgtcccg ccggagcagc 180

tgcggctcta cctcgcccac cgccgcctcc tcccggccga gccgtccccg ctgctgtcct 240

ccctccgggt ctcggcctcc tcctccctgc tactccacct ccccctgctc ggagggatga 300

ccggcccgac gacgaccccc gcggcacccc cgcccccgcc gccgccgtcg gcgcagccgc 360

ccgcccgccc cgcgcgctac gacttcctca actccaagcc gcccccgaac tacgtgggtc 420

tggggcgtgg cgccaccggg ttcaccaccc gttcggatat cgggccggcc cgcgcggcgc 480

ccgatctgcc tgaccggtcc gccgccgccg ccgccgcccc cgccgtcggg cgcggccgtg 540

ggaagccacc cggggacgac gacggcgacg acgatggcgg cgacgaggag aaggggtacg 600

acgagaacca gaagttcgac gagttcgagg gcaacgacgc cgggctgttc tccaacgccg 660

actacgacga cgacgaccgc gaggcggatg cggtctggga gagcatcgac cagaggatgg 720

actctcgccg gaaggatcgg cgggaggcgc ggctgaagca ggagatcgag aagtaccgtg 780

cttccaaccc taagatcacc gagcaattcg ctgatttgaa gcgtaagttg gtcgatttgt 840

cggcgcagga gtgggaaagc atacctgaaa ttggggacta ctcgctgcgc aacaagaaga 900

agcgatttga gagcttcgtt cccgtgccgg acaccctgct cgagaaggct cggcaggagc 960

aggagcatgt cacggcactg gatcccaaga gccgtgcagc tggtggcacc gagacgccat 1020

gggcgcagac tccggttacc gatctgacgg ctgtgggcga aggtcgtggc accgtgctct 1080

ccttgaagct ggacaggttg tcggattcgg tatctggtct tactgttgtt gatccaaagg 1140

gttacttgac ggacctgaaa agtatgaaga ttactagtga tgctgagatt tctgacatta 1200

aaaaggcgcg attgttgctt aagtcagtga cacagacaaa cccgaagcat ccaccaggat 1260

ggattgctgc tgctaggctt gaagaggttg ctggcaagct tcaggttgct cggcagctta 1320

tccagcgtgg ctgtgaggag tgccccacaa atgaggatgt ttgggtcgag gcatgccggc 1380

tggccagccc agacgaggca aaggcagtga ttgctagggg cgtgaaggca attcccaatt 1440

ctgtgaagct gtggttgcag gcagcaaagt tggaaactag tgatttgaat aagagcaggg 1500

ttttgagaaa agggttggaa cacattcctg attcagtcag actgtggaaa gcagtagtag 1560

agcttgcaaa tgaggaggat gcaagactgt tgcttcacag ggctgtggag tgctgcccac 1620

tccatgtgga actgtggctt gccctagcaa ggctggagac atatgaccaa gcaaagaagg 1680

tacttaacaa ggcaagagaa aagcttccta aggaacctgc catctggatt acagctgcaa 1740

agctggagga agctaatgga aacacccagt cagtaatcaa ggtgattgag agaagtataa 1800

aaactttaca gagagaagga ttggatattg acagggaggc atggctaaag gaagcagaag 1860

ctgctgagcg tgctggatct gtattgactt gccaggctat tgttaagagc actattggca 1920

ttggtgttga tgaggaagac agaaaacgca catgggttgc cgatgctgag gaatgcaaga 1980

agcgtggttc aattgagaca gcccgtgcca tctatgcgca tgcactcagt gtcttcgttt 2040

ccaagaagag tatttggctg aaagcggctc agcttgagaa gagccatgga accaaggagt 2100

ctctttataa tctcctcaga aaggctgtta cctacaatcc acgtgcagaa gttttatggc 2160

ttatgagtgc aaaggagaaa tggctggctg gagatgtccc ggctgcccga gccattcttc 2220

aggaagctta tgcttctctc cccaattcag aggagatctg gctagctgcc ttcaagcttg 2280

agtttgagaa caatgaacca gagagagcaa gaattctttt gtcaaaggcc agggaaagag 2340

gaggcactga gagggtctgg atgaaatctg cgattgttga aagggagtta gggaatgtag 2400

acgaagaaag gaagctgttg gaggaaggtc tgaagttatt cccctcattc ttcaagctgt 2460

ggttaatgct tggacaaatg gaagaccggc ttggccatgg atccaaggca aaggaggttt 2520

acgagaatgc actgaagcac tgcccgagtt gcatccctct ttggctctct ctagctaatc 2580

tagaggagaa gataaatggc ttgagcaagt cacgtgctgt cctcaccatg gcaagaaaga 2640

agaacccagc tacacctgaa ctctggcttg cagcagttag ggctgaattg agacatggga 2700

acaagaagga agctgatgct ctactagcca aggcattaca ggaatgcccg acaagtggta 2760

ttttgtgggc tgcagctata gagatggtgc cacgtcccca gcgtaaagca aagagctcag 2820

atgctataaa acgatgtgac catgatcccc atgtcattgc agctgtggcc aaacttttct 2880

ggcatgatag gaaggttgat aaagctagaa gttggttgaa tagagctgtt actcttgctc 2940

cagacattgg agatttttgg gccttgtact acaaatttga actgcaacat ggaaatgctg 3000

atacacaaaa ggatgtccta caaagatgtg ttgcagcaga accaaagcat ggagagagat 3060

ggcaagcaat aacaaaggct gttgagaact cacatctgtc aattgaggcc cttctgaaga 3120

aagctgtgtt ggctcttggc caggaagaaa atccaaatgc tgcagatccc tagtttgtct 3180

cacttttaac ttttgataag gtattgcaat ctgttatcat tatactcttc tgataaagaa 3240

ctttgctatt gtgttcccgt atttccatgc tttatgatgt ctcatattga aatgcttttc 3300

agtgtctatt ctattggtca gctataagat cttaatattt gagtatcatg taataaatat 3360

tgcgaagagt tcttaatatt tgagttgcaa ttaatttgtt tgagacaagc agcattatca 3420

tttattttgt tggttatcat taagtaaacc ttagcttaaa tctactaggt gcatgggtag 3480

tctaaaaata tgagttctgt atgtaaacta tgcagaatct gttcatgctg tattaatctg 3540

ctgtgcacat atgcccagtt atctatgaca tataaattat aatcatgttg atgctgtcat 3600

ggccttaaga ttgaaagaat atacttgttg ccttcagctt gatttatgtt ttggttgaag 3660

aattggtgtt gttttactcc ttgattgact gtatcacctt gactgaagta tttggggaat 3720

gtggcaattc tatgttgaag tgtgtcaatg ctaacattga ttactgagtt gcaagctgac 3780

tttctgctcc aaccaactct tgtgaatgtg catttttttg ccacaatagc ggtcagacta 3840

tttaattctg ctaagcaacc ctattctatc ctctcctgat tccaactgta gagcaataga 3900

agatcgataa aatgtctatg cggatcaaaa caccaccttt ggaagcataa tttttctttt 3960

tcttaccagt aattttgtgt ttctgtaaca aaacaagtaa ataacatatg ttactgctcg 4020

tctactgatt cacgggtatc ttttttagtt tcctatgtgc ttggattata attgtcttct 4080

tggatatccc cagactaaag ttttctttca actcctcagt ctggtcacag gtctaattct 4140

tctgcgcatg ctggcaatgg aatatataga gaaagaaaca cagttgggta gtgaactggg 4200

atggactcga gctgcgacct cgtgattctg tgtaccaccg aactgattgc tgcactcctc 4260

caaccatgaa gccttacctg aagaagaatc aggcatgcga ttactacaat ttgtatcggc 4320

gatcaccagt taaaacctgt atcgtttgta tgccctaatt gccagcaata cttctgtact 4380

accatgagca tgtttattcc tccagatgca taccacaaat tcttagatgg gtgtatttgc 4440

tagccgcgac ttgggatgat gtaatttttc ttgggttcgg tttattctca gagcactggc 4500

gtctgtatct acgactgtaa gatctgcctg aatgtcggct taatatatga gataatgccc 4560

catttttcag cacaggctgt gagcatttct tc 4592

<210> 2

<211> 3117

<212> DNA

<213> tas1 mutant (transparent-aspect and soft 1)

<400> 2

atggtgttcg tccgcgcgcc ggacgggagg acccaccacg tcgacctcga cccctccacc 60

gccacgctcg ccgacctcac ggcctccgcc tcccgcgtct gcggcggcgt cccgccggag 120

cagctgcggc tctacctcgc ccaccgccgc ctcctcccgg ccgagccgtc cccgctgctg 180

tcctccctcc gggtctcggc ctcctcctcc ctgctactcc acctccccct gctcggaggg 240

atgaccggcc cgacgacgac ccccgcggca cccccgcccc cgccgccgcc gtcggcgcag 300

ccgcccgccc gccccgcgcg ctacgacttc ctcaactcca agccgccccc gaactacgtg 360

ggtctggggc gtggcgccac cgggttcacc acccgttcgg atatcgggcc ggcccgcgcg 420

gcgcccgatc tgcctgaccg gtccgccgcc gccgccgccg cccccgccgt cgggcgcggc 480

cgtgggaagc cacccgggga cgacgacggc gacgacgatg gcggcgacga ggagaagggg 540

tacgacgaga accagaagtt cgacgagttc gagggcaacg acgccgggct gttctccaac 600

gccgactacg acgacgacga ccgcgaggcg gatgcggtct gggagagcat cgaccagagg 660

atggactctc gccggaagga tcggcgggag gcgcggctga agcaggagat cgagaagtac 720

cgtgcttcca accctaagat caccgagcaa ttcgctgatt tgaagcgtaa gttggtcgat 780

ttgtcggcgc aggagtggga aagcatacct gaaattgggg actactcgct gcgcaacaag 840

aagaagcgat ttgagagctt cgttcccgtg ccggacaccc tgctcgagaa ggctcggcag 900

gagcaggagc atgtcacggc actggatccc aagagccgtg cagctggtgg caccgagacg 960

ccatgggcgc agactccggt taccgatctg acggctgtgg gcgaaggtcg tggcaccgtg 1020

ctctccttga agctggacag gttgtcggat tcggtatctg gtcttactgt tgttgatcca 1080

aagggttact tgacggacct gaaaagtatg aagattacta gtgatgctga gatttctgac 1140

attaaaaagg cgcgattgtt gcttaagtca gtgacacaga caaacccgaa gcatccacca 1200

ggatggattg ctgctgctag gcttgaagag gttgctggca agcttcaggt tgctcggcag 1260

cttatccagc gtggctgtga ggagtgcccc acaaatgagg atgtttgggt cgaggcatgc 1320

cggctggcca gcccagacga ggcaaaggca gtgattgcta ggggcgtgaa ggcaattccc 1380

aattctgtga agctgtggtt gcaggcagca aagttggaaa ctagtgattt gaataagagc 1440

agggttttga gaaaagggtt ggaacacatt cctgattcag tcagactgtg gaaagcagta 1500

gtagagcttg caaatgagga ggatgcaaga ctgttgcttc acagggctgt ggagtgctgc 1560

ccactccatg tggaactgtg gcttgcccta gcaaggctgg agacatatga ccaagcaaag 1620

aaggtactta acaaggcaag agaaaagctt cctaaggaac ctgccatctg gattacagct 1680

gcaaagctgg aggaagctaa tggaaacacc cagtcagtaa tcaaggtgat tgagagaagt 1740

ataaaaactt tacagagaga aggattggat attgacaggg aggcatggct aaaggaagca 1800

gaagctgctg agcgtgctgg atctgtattg acttgccagg ctattgttaa gagcactatt 1860

ggcattggtg ttgatgagga agacagaaaa cgcacatggg ttgccgatgc tgaggaatgc 1920

aagaagcgtg gttcaattga gacagcccgt gccatctatg cgcatgcact cagtgtcttc 1980

gtttccaaga agagtatttg gctgaaagcg gctcagcttg agaagagcca tggaaccaag 2040

gagtctcttt ataatctcct cagaaaggct gttacctaca atccacgtgc agaagtttta 2100

tggcttatga gtgcaaagga gaaatggctg gctggagatg tcccggctgc ccgagccatt 2160

cttcaggaag cttatgcttc tctccccaat tcagaggaga tctggctagc tgccttcaag 2220

cttgagtttg agaacaatga accagagaga gcaagaattc ttttgtcaaa ggccagggaa 2280

agaggaggca ctgagagggt ctggatgaaa tctgcgattg ttgaaaggga gttagggaat 2340

gtagacgaag aaaggaagct gttggaggaa ggtctgaagt tattcccctc attcttcaag 2400

ctgtggttaa tgcttggaca aatggaagac cggcttggcc atggatccaa ggcaaaggag 2460

gtttacgaga atgcactgaa gcactgcccg agttgcatcc ctctttggct ctctctagct 2520

aatctagagg agaagataaa tggcttgagc aagtcacgtg ctgtcctcac catggcaaga 2580

aagaagaacc cagctacacc tgaactctgg cttgcagcag ttagggctga attgagacat 2640

gggaacaaga aggaagctga tgctctacta gccaaggcat tacaggaatg cccgacaagt 2700

ggtattttgt gggctgcagc tatagagatg gtgccacgtc cccagcgtaa agcaaagagc 2760

tcagatgcta taaaacgatg tgaccatgat ccccatgtca ttgcagctgt ggccaaactt 2820

ttctggcatg ataggaaggt tgataaagct agaagttggt tgaatagagc tgttactctt 2880

gctccagaca ttggagattt ttgggccttg tactacaaat ttgaactgca acatggaaat 2940

gctgatacac aaaaggatgt cctacaaaga tgtgttgcag cagaaccaaa gcatggagag 3000

agatggcaag caataacaaa ggctgttgag aactcacatc tgtcaattga ggcccttctg 3060

aagaaagctg tgttggctct tggccaggaa gaaaatccaa atgctgcaga tccctag 3117

<210> 3

<211> 1038

<212> PRT

<213> tas1 mutant (transparent-apearance and soft 1)

<400> 3

Met Val Phe Val Arg Ala Pro Asp Gly Arg Thr His His Val Asp Leu

1 5 10 15

Asp Pro Ser Thr Ala Thr Leu Ala Asp Leu Thr Ala Ser Ala Ser Arg

20 25 30

Val Cys Gly Gly Val Pro Pro Glu Gln Leu Arg Leu Tyr Leu Ala His

35 40 45

Arg Arg Leu Leu Pro Ala Glu Pro Ser Pro Leu Leu Ser Ser Leu Arg

50 55 60

Val Ser Ala Ser Ser Ser Leu Leu Leu His Leu Pro Leu Leu Gly Gly

65 70 75 80

Met Thr Gly Pro Thr Thr Thr Pro Ala Ala Pro Pro Pro Pro Pro Pro

85 90 95

Pro Ser Ala Gln Pro Pro Ala Arg Pro Ala Arg Tyr Asp Phe Leu Asn

100 105 110

Ser Lys Pro Pro Pro Asn Tyr Val Gly Leu Gly Arg Gly Ala Thr Gly

115 120 125

Phe Thr Thr Arg Ser Asp Ile Gly Pro Ala Arg Ala Ala Pro Asp Leu

130 135 140

Pro Asp Arg Ser Ala Ala Ala Ala Ala Ala Pro Ala Val Gly Arg Gly

145 150 155 160

Arg Gly Lys Pro Pro Gly Asp Asp Asp Gly Asp Asp Asp Gly Gly Asp

165 170 175

Glu Glu Lys Gly Tyr Asp Glu Asn Gln Lys Phe Asp Glu Phe Glu Gly

180 185 190

Asn Asp Ala Gly Leu Phe Ser Asn Ala Asp Tyr Asp Asp Asp Asp Arg

195 200 205

Glu Ala Asp Ala Val Trp Glu Ser Ile Asp Gln Arg Met Asp Ser Arg

210 215 220

Arg Lys Asp Arg Arg Glu Ala Arg Leu Lys Gln Glu Ile Glu Lys Tyr

225 230 235 240

Arg Ala Ser Asn Pro Lys Ile Thr Glu Gln Phe Ala Asp Leu Lys Arg

245 250 255

Lys Leu Val Asp Leu Ser Ala Gln Glu Trp Glu Ser Ile Pro Glu Ile

260 265 270

Gly Asp Tyr Ser Leu Arg Asn Lys Lys Lys Arg Phe Glu Ser Phe Val

275 280 285

Pro Val Pro Asp Thr Leu Leu Glu Lys Ala Arg Gln Glu Gln Glu His

290 295 300

Val Thr Ala Leu Asp Pro Lys Ser Arg Ala Ala Gly Gly Thr Glu Thr

305 310 315 320

Pro Trp Ala Gln Thr Pro Val Thr Asp Leu Thr Ala Val Gly Glu Gly

325 330 335

Arg Gly Thr Val Leu Ser Leu Lys Leu Asp Arg Leu Ser Asp Ser Val

340 345 350

Ser Gly Leu Thr Val Val Asp Pro Lys Gly Tyr Leu Thr Asp Leu Lys

355 360 365

Ser Met Lys Ile Thr Ser Asp Ala Glu Ile Ser Asp Ile Lys Lys Ala

370 375 380

Arg Leu Leu Leu Lys Ser Val Thr Gln Thr Asn Pro Lys His Pro Pro

385 390 395 400

Gly Trp Ile Ala Ala Ala Arg Leu Glu Glu Val Ala Gly Lys Leu Gln

405 410 415

Val Ala Arg Gln Leu Ile Gln Arg Gly Cys Glu Glu Cys Pro Thr Asn

420 425 430

Glu Asp Val Trp Val Glu Ala Cys Arg Leu Ala Ser Pro Asp Glu Ala

435 440 445

Lys Ala Val Ile Ala Arg Gly Val Lys Ala Ile Pro Asn Ser Val Lys

450 455 460

Leu Trp Leu Gln Ala Ala Lys Leu Glu Thr Ser Asp Leu Asn Lys Ser

465 470 475 480

Arg Val Leu Arg Lys Gly Leu Glu His Ile Pro Asp Ser Val Arg Leu

485 490 495

Trp Lys Ala Val Val Glu Leu Ala Asn Glu Glu Asp Ala Arg Leu Leu

500 505 510

Leu His Arg Ala Val Glu Cys Cys Pro Leu His Val Glu Leu Trp Leu

515 520 525

Ala Leu Ala Arg Leu Glu Thr Tyr Asp Gln Ala Lys Lys Val Leu Asn

530 535 540

Lys Ala Arg Glu Lys Leu Pro Lys Glu Pro Ala Ile Trp Ile Thr Ala

545 550 555 560

Ala Lys Leu Glu Glu Ala Asn Gly Asn Thr Gln Ser Val Ile Lys Val

565 570 575

Ile Glu Arg Ser Ile Lys Thr Leu Gln Arg Glu Gly Leu Asp Ile Asp

580 585 590

Arg Glu Ala Trp Leu Lys Glu Ala Glu Ala Ala Glu Arg Ala Gly Ser

595 600 605

Val Leu Thr Cys Gln Ala Ile Val Lys Ser Thr Ile Gly Ile Gly Val

610 615 620

Asp Glu Glu Asp Arg Lys Arg Thr Trp Val Ala Asp Ala Glu Glu Cys

625 630 635 640

Lys Lys Arg Gly Ser Ile Glu Thr Ala Arg Ala Ile Tyr Ala His Ala

645 650 655

Leu Ser Val Phe Val Ser Lys Lys Ser Ile Trp Leu Lys Ala Ala Gln

660 665 670

Leu Glu Lys Ser His Gly Thr Lys Glu Ser Leu Tyr Asn Leu Leu Arg

675 680 685

Lys Ala Val Thr Tyr Asn Pro Arg Ala Glu Val Leu Trp Leu Met Ser

690 695 700

Ala Lys Glu Lys Trp Leu Ala Gly Asp Val Pro Ala Ala Arg Ala Ile

705 710 715 720

Leu Gln Glu Ala Tyr Ala Ser Leu Pro Asn Ser Glu Glu Ile Trp Leu

725 730 735

Ala Ala Phe Lys Leu Glu Phe Glu Asn Asn Glu Pro Glu Arg Ala Arg

740 745 750

Ile Leu Leu Ser Lys Ala Arg Glu Arg Gly Gly Thr Glu Arg Val Trp

755 760 765

Met Lys Ser Ala Ile Val Glu Arg Glu Leu Gly Asn Val Asp Glu Glu

770 775 780

Arg Lys Leu Leu Glu Glu Gly Leu Lys Leu Phe Pro Ser Phe Phe Lys

785 790 795 800

Leu Trp Leu Met Leu Gly Gln Met Glu Asp Arg Leu Gly His Gly Ser

805 810 815

Lys Ala Lys Glu Val Tyr Glu Asn Ala Leu Lys His Cys Pro Ser Cys

820 825 830

Ile Pro Leu Trp Leu Ser Leu Ala Asn Leu Glu Glu Lys Ile Asn Gly

835 840 845

Leu Ser Lys Ser Arg Ala Val Leu Thr Met Ala Arg Lys Lys Asn Pro

850 855 860

Ala Thr Pro Glu Leu Trp Leu Ala Ala Val Arg Ala Glu Leu Arg His

865 870 875 880

Gly Asn Lys Lys Glu Ala Asp Ala Leu Leu Ala Lys Ala Leu Gln Glu

885 890 895

Cys Pro Thr Ser Gly Ile Leu Trp Ala Ala Ala Ile Glu Met Val Pro

900 905 910

Arg Pro Gln Arg Lys Ala Lys Ser Ser Asp Ala Ile Lys Arg Cys Asp

915 920 925

His Asp Pro His Val Ile Ala Ala Val Ala Lys Leu Phe Trp His Asp

930 935 940

Arg Lys Val Asp Lys Ala Arg Ser Trp Leu Asn Arg Ala Val Thr Leu

945 950 955 960

Ala Pro Asp Ile Gly Asp Phe Trp Ala Leu Tyr Tyr Lys Phe Glu Leu

965 970 975

Gln His Gly Asn Ala Asp Thr Gln Lys Asp Val Leu Gln Arg Cys Val

980 985 990

Ala Ala Glu Pro Lys His Gly Glu Arg Trp Gln Ala Ile Thr Lys Ala

995 1000 1005

Val Glu Asn Ser His Leu Ser Ile Glu Ala Leu Leu Lys Lys Ala Val

1010 1015 1020

Leu Ala Leu Gly Gln Glu Glu Asn Pro Asn Ala Ala Asp Pro

1025 1030 1035

Claims (10)

1. AlleleDu1 ^ΔA12 Characterised in that the allele isDu1 ^ΔA12 Is wild riceDu1The 361 st to 363 th nucleotides of the 1 st exon of the coding region of the gene are deleted.

2. The allele according to claim 1Du1 ^ΔA12 Characterized in that said allele isDu1 ^ΔA12 The nucleotide sequence of (A) is shown as SEQ ID No.1Shown in the figure.

3. The allele according to claim 1 or 2Du1 ^ΔA12 Characterised in that the allele isDu1 ^ΔA121 The sequence of the coding region is shown as SEQ ID No. 2.

4. The allele according to any one of claims 1 to 3Du1 ^ΔA12 The coded protein is characterized in that the amino acid sequence of the coded protein is shown as SEQ ID No. 3.

5. An expression cassette, recombinant vector or cell comprising the allele of any one of claims 1 to 3.

6. The allele according to any one of claims 1 to 3Du1 ^ΔA12 The protein of claim 4, the expression cassette of claim 5, the recombinant vector or the cell, and the use thereof in rice cross breeding and variety improvement.

7. Use according to claim 6, wherein the use comprises controlling the apparent transparency, and/or low amylose content of rice.

8. A method for obtaining rice with a low amylose content and a transparent appearance, comprising the steps of:

1) allowing rice to contain the allele according to any one of claims 1 to 3Du1 ^ΔA12 (ii) a Or

2) Expressing the allele of claim 4 in riceDu1 ^ΔA12 The encoded protein.

9. The method according to claim 8, characterized in that it comprises a transgenic, crossing, backcrossing or asexual propagation step.

10. Method for identifying rice, whichWherein the rice plant comprises the allele according to any one of claims 1 to 3Du1 ^ΔA12 The rice of (1), rice expressing the protein of claim 4 or rice obtained by the method of claim 8 or 9, comprising the steps of:

1) identifying whether the rice comprises an allele according to any one of claims 1 to 3; or the like, or, alternatively,

2) identifying whether the rice expresses the protein of claim 4.

Images