NL2032897B1 - GENE Gmygl2 RELATED TO SOYBEAN PLANT HEIGHT AND LEAF COLOR, INSERTION-DELETION (InDel) MARKER OF GENE Gmygl2, AND USE THEREOF - Google Patents

Abstract

The present disclosure relates to a gene Gmygl2 related to soybean plant height and leaf color, an insertion-deletion (InDel) marker of the Gmygl2 gene, and use thereof. In the present disclosure, an F2 population is constructed with GL11 as a male parent and a mutant ygl2 with a tall stem and a yellow-green leaf as a female parent, individuals with a tall stem and a yellow-green leaf and individuals with a short stem and a green leaf are selected in the F2 population to construct a yellow-green leaf pool and a green leaf pool, an interval linked with a yellow-green leaf phenotype is located using a bulked segregant analysis (BSA) method and a single sequence repeat (SSR) marker technology, to obtain the gene Gmygl2 related to soybean leaf color, with a nucleotide sequence shown in SEQ ID NO: 1.

Description

GENE Gmygl2 RELATED TO SOYBEAN PLANT HEIGHT AND LEAF

COLOR, INSERTION-DELETION (InDel) MARKER OF GENE Gmygi2, AND

USE THEREOF

TECHNICAL FIELD

[1] The present disclosure belongs to the technical field of crop genetic breeding and molecular biology, and particularly relates to a gene Gmyg!2 related to soybean plant height and leaf color, an insertion-deletion (InDel) marker of the Gmyg/2 gene, and use thereof.

BACKGROUND ART

[2] Soybean is a main source of edible vegetable oils and vegetable proteins, providing the vegetable proteins and fats for human beings. The complex gene structure, low-efficiency genetic transformation and many other factors limit the progress of soybean functional genomics researches.

[3] Soybean leaf color mutants are important materials for the study of gene functions related to regulation on chlorophyll biosynthesis, chloroplast development, and photosynthesis; moreover, these mutants are of great significance for studying soybean photosynthesis, improving soybean photosynthetic efficiency, and promoting soybean production; in addition, these mutants are also important for the authenticity identification of hybrids. Therefore, it is of great theoretical significance to explore stress resistance genes related to photosynthesis, and to explore structures and functions of the genes, for elucidating a regulation mechanism of the photosynthesis under adversity and cultivating soybeans with a high light efficiency.

SUMMARY

[4] A purpose of the present disclosure is to provide a gene Gmyg?2 related to soybean plant height and leaf color, an insertion-deletion (InDel) marker of the

Gmygl2 gene, and use thereof, thereby rapidly screening and identifying a leaf color phenotype and a plant height phenotype of the soybean, to improve a breeding efficiency of soybeans with a high light efficiency.

[5] In the present disclosure, an F2 population is constructed with GL11 as a male parent and a mutant yg/2 with a yellow-green leaf as a female parent; individuals with a yellow-green leaf and individuals with a green leaf are selected in the F2 population to construct a yellow-green leaf pool and a green leaf pool; an interval linked with a yellow-green leaf phenotype is located using a bulked segregant analysis (BSA) method and a single sequence repeat (SSR) marker technology, to obtain the gene

Gmygl2 related to soybean leaf color, with a nucleotide sequence shown in SEQ ID

NO: 1.

[6] In the present disclosure, with a sequence difference between wild soybean

GL11 and a mutant yg/2, primers are designed according to conserved sequences at both ends of an InDel site, such that an InDel marker is successfully developed. The

InDel marker can identify leaf color and plant height phenotypes of the soybean by simple DNA extraction and PCR-specific amplification. Accordingly, a soybean material with a yellow-green leaf color and a desirable plant height can be obtained to accelerate a breeding process.

BRIEF DESCRIPTION OF THE DRAWINGS

[7] To explain the technical solutions in embodiments of the present disclosure or in the prior art more clearly, the accompanying drawings required in the embodiments will be described below in brief.

[8] FIG. 1 shows a leaf color phenotype diagram of seedling stage plants of GL11 and yg/2;

[9] FIG. 2 shows a plant height phenotype diagram of mature stage plants of the

GL11 and the vg/2;

[10] FIG. 3 shows an agronomic character analysis diagram of a yellow-green leaf mutant yg/2; [MI] FIG. 4 shows a linkage map of a gene (;imyg/2 on chromosome 2;

[12] FIG. 5 shows a gene expression characteristic in a leaf between parents in a location interval of the gene Gmygl2,

[13] FIG. 6 shows an alignment diagram of gene sequences and protein sequences of the GL 11 and the yg/2;

[14] FIG. 7 shows a comparison chart of the plant height and the number of nodes of the seedling stage plants of the GL11 and the yg/2;

[15] FIG. 8 shows a cell structure diagram of an internode length of the seedling stage plants of the GL 11 and the yg/2;

[16] FIG. 9 shows a schematic diagram of a leaf chlorophyll content of the seedling stage plants of the GL 11 and the yg/2;

[17] FIG. 10 shows an ultrastructure diagram of leaf chlorophyll of the seedling stage plants of the GL11 and the vg/2;

[18] FIG. 11 shows a verification electropherogram of 45 copies of GL11xvg/2 (F2:3) known genotype material and parents GL11 and yg/2 on a molecular marker;

[19] FIG. 12 shows a verification electropherogram of 40 copies of known genotype materials of domestic and foreign germplasm resources and the parents GL11 and yg/2 on the molecular marker;

[20] FIG. 13 shows a yellow-green leaf phenotype after CRISPR/Cas9 gene knockout; and

[21] FIG. 14 shows an alignment diagram of amino acid sequences of GL11 (SEQ

ID NO: 3), vgi2 (SEQ ID NO: 5), and a gene-edited seedling YGL2-SP1 (SEQ ID

NO:10).

DETAILED DESCRIPTION OF THE EMBODIMENTS

[22] The present disclosure provides a gene Gmyg/2 related to soybean plant height and leaf color, having a nucleotide sequence shown in SEQ ID NO: 1.

[23] In the present disclosure, an F2 population is constructed with GL11 as a male parent and a mutant yg/2 with a tall stem and a yellow-green leaf as a female parent; individuals with a tall stem and a yellow-green leat and individuals with a short stem and a green leaf are selected in the F2 population to construct a yellow-green leaf pool and a green leaf pool; an interval linked with a yellow-green leaf phenotype is located using a bulked segregant analysis (BSA) method and a single sequence repeat (SSR) marker technology, to obtain the gene Gmygi2 related to soybean leaf color. The soybean leaf color-related gene Gmyg/2 is located on chromosome 2 of the soybean gene (accession number Glyma. 02g304700), with a nucleotide sequence shown in SEQ

ID NO: I. The gene Gmyg?/2 related to soybean plant height and leaf color has a nucleotide sequence length of 7,475 bp, including 9 exons and 8 introns; and the gene

Gmygl2 has a total CDS length of 990 bp, with a specific nucleotide sequence shown in SEQ ID NO: 2. A protein encoded by a CDS gene has an amino acid sequence as shown in SEQ ID NO: 3, with a total of 329 amino acids. The results in examples show that the gene Gmyg/2 is a functional gene that causes plant internode elongation and yellow-green leaf phenotypes, and can be used to screen a yellow-green leaf gene with a desirable plant height and provide uses thereof.

[24] A nucleotide sequence formed by conducting substitution, deletion, or addition of bases on the nucleotide sequence shown in SEQ ID NO: 1 and having a same function as the nucleotide sequence shown in SEQ ID NO: 1, the Gmyg/2 gene, or the protein encoded by the Gmyg?2 gene is mainly used for identifying the color of soybean leaves, preparing products capable of identifying the color of soybean leaves, assisting in identifying the color of soybean leaves, preparing products capable of assisting in identifying the color of soybean leaves, and soybean breeding, which are all within the scope of protection of the present disclosure.

[25] The present disclosure further provides use of the gene Gmygl2 related to soybean plant height and leaf color or the protein in any one of B1 to B5:

[26] BI, identifying a leaf color and a plant height of a soybean;

[27] B2, preparing a product capable of identifying the leaf color and the plant height of the soybean;

[28] B3, assisting in identifying the leaf color and the plant height of the soybean;

[29] B4, preparing a product capable of assisting in identifying the leaf color and the plant height of the soybean; and

[30] B5, soybean breeding. [BI] The present disclosure further provides an InDel marker linked to the gene

Gmiygl2 related to soybean plant height and leaf color, where primers of the InDel marker includes an upstream primer InDel-105-F with a nucleotide sequence shown in

SEQ ID NO: 6 and a downstream primer InDel-105-R with a nucleotide sequence shown in SEQ ID NO: 7.

[82] In the present disclosure, the InDel marker is located on chromosome 2 of the soybean genome, specifically at 1.167 Mb upstream of the yg/2 gene on chromosome 2.

With a sequence difference between wild soybean GL 11 and a mutant yg/2, primers are designed according to conserved sequences at both ends of an InDel site, such that an

InDel marker is successfully developed to identify the leaf color phenotype of the soybean.

[33] The nucleotide sequences with an equivalent function formed by conducting substitution, deletion, or addition of bases on the nucleotide sequences shown in SEQ

ID NO: 6 and SEQ ID NO: 7, or the nucleotide sequences with the equivalent function and as derivatives of the nucleotide sequences shown in SEQ ID NO: 6 and SEQ ID

NO: 7 should all fall within the protection scope of the present disclosure.

[34] The present disclosure further provides use of the InDel marker in any one of

Bl to BS: 5 [35] BI, identifying a leaf color and a plant height of a soybean;

[36] B2, preparing a product capable of identifying the leaf color and the plant height of the soybean;

[37] B3, assisting in identifying the leaf color and the plant height of the soybean;

[38] B4, preparing a product capable of assisting in identifying the leaf color and the plant height of the soybean; and

[39] BS, soybean breeding.

[40] The present disclosure further provides a method for screening a soybean with a vellow-green leaf color and an elongated internode, including the following steps: conducting PCR amplification on a genomic DNA of a soybean to be screened using the primers of the InDel marker, and selecting a material with a PCR amplification product of 149 bp, to obtain the soybean with a yellow-green leaf color and a desirable plant height.

[41] In the present disclosure, a PCR amplification system includes preferably: 1 uL to 2 uL of a DNA template (80 ng to 150 ng), 0.6 uL of each of the upstream primer

InDel-105-F and the downstream primer InDel-105-R, 2 uL of a 10*xEasy Taq Buffer (Mg** free), 2 uL of dNTPs (2.5 mM), 0.2 pL of Easy Taq Polymerase (TransGen,

API111), and adding ddH;O to make up to 20 pL. A reaction procedure of the PCR amplification includes preferably: initial denaturation at 95°C for 5 min; denaturation at 95°C for 30 sec, annealing at (Tm-5)°C for 30 sec, and extension at 72°C (1 min/kb), conducting 35 cycles; and extension at 72°C for 8 min. There is no special limitation on a method for obtaining the genomic DNA of the soybean to be selected, and methods well known in the art can be used, such as a CTAB method.

[42] In order to further illustrate the present disclosure, the technical solutions provided by the present disclosure are described in detail below in connection with examples, but these examples should not be understood as limiting the claimed scope of the present disclosure.

[43] Example 1

[44] 1. Construction of genetic populations

[45] In the Beijing Shunyi Experimental Base of the Chinese Academy of

Agricultural Sciences, GL11 was used as a male parent, and a yellow-green leaf mutant vg/2 was used as a female parent (WANG Hao-Rang, ZHANG Yong, YU

Chun-Miao, DONG Quan-Zhong, LI Wei-Wei, HU Kai-Feng, ZHANG Ming-Ming,

XUE Hong, YANG Meng-Ping, SONG Ji-Ling, WANG Lei, YANG Xing-Yong, and

QIU Li-Juan. Fine mapping of yellow-green leaf gene (ygl2) in soybean (Glycine max

L) IJ]. Acta Agronomica Sinica, 2022, 48 (04): 791-800), and a hybrid combination was configured to obtain hybrid grains. In the summer of 2017, F1 generation plants were propagated and harvested at the same location; F2 generation populations were planted in the same location in 2018; in 2019, F3 generation populations were continued to be planted and phenotyped to verify a F2 population genotype.

[46] 2. Fine localization of soybean gene Gmiygl2

[47] In an F2 population of the combination yg/2xGL11, 45 individuals with tall stem and yellow-green leat and individuals with dwarf stem and green leaf were selected to construct a yellow-green leaf pool and a green leaf pool; deep resequencing of the DNA pools of the two materials was conducted on a Hiseq2500 platform. Taking a Williams 82 genome sequence as a reference, single nucleotide polymorphism sites between genome sequences of the two pools were discovered, and yellow-green leaf-related candidate intervals were screened by an ED (Euler distance) association algorithm for subsequent analysis. The genetic analysis of leaf color was conducted on the F2 population to verify an inheritance law of the leaf color; a single-plant genomic

DNA of the F2 population of yg/2xGL11 was extracted, and band patterns of the offspring individuals were amplified using associated markers screened by sequencing, and the intervals linked to the yellow-green leaf phenotype were localized.

[48] In the autumn of 2018, in Shunyi, Beijing, the number of grains per plant, grain weight, 100-grain weight, protein, fat, plant height and node number of yg/2 and

GL11 at maturity were investigated. As shown in FIGs. 1 to 3, where FIGs. | and 2 showed the growth of GL11 and yg/2 from left to right, respectively. According to

FIGs. 1 to 3, compared with GL11, in the yellow-green leat mutant yg/2, except for plant height, 100-grain weight and protein content, the number of grains per plant and the grain weight per plant decreased significantly.

[49] 3. The genetic analysis was conducted using the constructed GL11xyg/2, and the results were shown in Table 1.

[50] Table 1 Segregation of genetic F2 offspring of GL11xyg/2

[51]

Population GL11xygl2 (F2:3)

Phenotype Total Green Green-Yellow Pa p-Value

No. 567 412 155 1.529 0.1988

[82] With the F2 segregated population assembled by vg/2 and GLI, the yellow-green leaf gene was localized on chromosome 2 by the BSA method, which was between markers 02 55 and 02 130 with an interval size of about 1.167 Mb. In the interval, 61 SSR markers were selected for polymorphism screening among parents, and 24 SSR markers with differences among parents were obtained; with these markers, 155 yellow-green F2 individual plants were genotyped, a genetic map was drawn combined with the phenotype of the F3 line to narrow a localized interval, and the localized interval was reduced to 1.167 Mb; the localized interval was mapped to a soybean reference genome to find candidate genes, and the interval included a total of 170 genes (FIG. 4).

[53] By the markers 02 55 and 02 130, 1440 F3 genotypes in the population were identified, and 228 individual plants with different genotypes between the two markers were screened. Another 53 SSR markers were selected between the 02 55 and 02 130, 12 of which were polymorphic between yg/2 and GLII; with these 12 pairs of polymorphism markers, genotypes of the above 228 mutant individuals were analyzed, and 6 key crossover individuals were identified. After recombination analysis, the target gene Gmygl2 was localized between 02 104 and 02 107 (FIG. 4), with an interval size of 56.1 Kb.

[54] By a soybean genome annotation website (https://soybase.org/), a 56.1 kb interval where the Gmyvg/2 gene was located was analyzed in the reference genome, as shown in Table 2.

[55] Table 2 Gene annotation information in the localized interval

[56]

Gene Gene annotation information

Genel Transcriptional regulator sterile apetala-like protein

Creme? Unknown protein

Genel Unknown protein

CGiered F-box-like protein

Genes DNA; homolog subfamily c member protein

Gene6 Non-specific protein-tyrosine kinase/Cytoplasmic protein tyrosine kinase

Gene? Nuclear pore complex protein Nupl133 (NUP133)

Gene8 Calcium-transporting atpase / calcium-translocating p-type protein

Gene Phytochromobilin synthase

[57] The results showed that there were 9 annotated genes in the localized interval, including 7 known protein-encoding genes and 2 genes with unknown functions, and no reported soybean leaf color-related genes had been found in the interval, indicating that the Gmyg/2 (Gene9) leaf color gene localized in the present disclosure was a new gene.

[58] 4. Through a fluorescence quantitative PCR technology, gene expression differences of genes in the candidate interval were compared between leaf tissues of the yellow-green leaf mutant yg/2 and the green leaf near-isogenic line GL11, and the results were shown in FIG. 5. According to FIG. 5, it was seen that Gene9 had the maximum expression level between the two parents with an extremely significant difference, such that the Gene9 (Gmygi2) was a strong candidate gene for soybean leaf phenotypes.

[59] Example 2

[60] Isolation and structural analysis of gene Gmvg/2 in soybean

[61] A total RNA was extracted from a wild-type soybean variety GL11; the total

RNA was used as a template, and PCR amplification was conducted with a forward primer (having a nucleotide sequence shown in SEQ ID NO: 8, 5-ATGGGTTTTAGAATTAGCGGT-3") and a reverse primer (having a nucleotide sequence shown in SEQ ID NO: 9, 5'-CCTAAAATATTCAATAAATTCTCCTC-3') separately. A cDNA fragment of the gene (Gmyg/2 was obtained with a length of 990 bp (SEQ ID NO: 2); a genomic DNA from young leaves of the soybean variety GL11 was extracted, and the Gmyg/2 genomic fragment was amplified using the genomic DNA as a template. The gene encoded 329 amino acids (SEQ ID NO: 3) (FIG. 6).

[62] Example 3

[63] 1. Isolation of soybean gene Ginygl2 mutant

[64] The yg/2 mutant was a natural mutation, with a gene (Gmyg/2 sequence shown in SEQ ID NO: 4; exons of the gene Gmvg/2 mutant had base deletion, and frameshift mutations led to premature termination of protein translation; a protein encoded by this gene had an amino acid sequence mutation shown in SEQ ID NO: 5, with a total of 98 amino acids (FIG. 6).

[65] 2. Phenotypic characteristics of gene (Gmyg/2 in soybean

[66] From 2016 to 2018, field phenotype observation and identification of the yellow-green leaf mutant yg/2 and the wild-type soybean variety GL11 during the entire growth period were conducted at the Beijing Changping Experiment Station of the Institute of Crop Science, Chinese Academy of Agricultural Sciences (referring to the "Soybean Germplasm Description Specifications and Data Standards" edited by

Lijuan Qiu et al).

[67] 3. Physiological characteristics of gene Gmygl2 in soybean

[68] (1) At a seedling stage (V4), the wild-type GL11 and the yellow-green leaf mutant vg/2 that had consistent growth were selected, and the pigment contents of upper, middle, and lower leaves of the plant were determined (Plant Physiology

Experiment Guide [M]. Higher Education Press, Junfeng Gao Editor-in-Chief, 2006); the formulas were as follows:

[69] Chlorophyll a concentration Ca=13.95xDees to 6.88% Deas

[70] Chlorophyll b concentration Cb=24.96xDe49 to 7.32% Des

[71] Carotenoid concentration Cx=(1000xDy70 to 2.05xCa - 114xCb)/245

[72] Chloroplast pigment content (mg/g) = pigment concentration (C) * extract volume (mL)/sample fresh or dry weight (g).

[73] (2) Referring to "Practical Bio-electron Microscopy" (Practical Bio-electron

Microscopy [M]. Liaoning Science and Technology Press, edited by Junan Lin et al, 1989), ultrathin sections of the chloroplast were prepared, observed by an electron microscope, and then photographed for preservation.

[74] The detection results of physiological characteristics of the gene Gmyg/2 in soybean were shown in FIGs. 1 to 2 and 8 to 9. It was seen that the phenotype of the yellow-green leaf mutant yg/2 had yellow-green leaves, the plant showed a yellow-green phenotype of leaves from a seedling stage. Compared with the wild type, the plant had a taller plant, a larger grain size, and a higher 100-grain weight at the mature stage (FIG. 1 and FIG. 2), the internodes were elongated, but the number of nodes remained unchanged. (FIG. 7 and FIG. 8), which might be caused by plant height phenotype; a chlorophyll content of leaves from yg/2 plants was significantly lower than that of the wild-type GL 11 (FIG. 9); compared with wild type, the yg/2 had significantly lower chloroplast grana stacking than the GL11, with reduced chloroplast grana lamellae and irregular size and shape (FIG. 10), which might be responsible for the yellow-green leaf phenotype.

[75] Example 4

[76] Development and utilization of functional markers of gene Gmygi2 in soybean

[77] (1) With a sequence difference between wild soybean GL 11 and a mutant yg/2, primers were designed according to conserved sequences at both ends of an InDel site, such that an InDel marker was successfully developed to identify populations (Table 3).

[78] Table 3 Primer sequences of InDel marker

[79]

Name of marker Primer

F:AAGAGTTTTACTATGACTATGT (SEQ ID NO: 6)

D103

R:ATAAATACAGACAAAGTCAAAT (SEQ ID NO: 7)

[80] The genotypes of 155 F2 recessive individuals (yellow-green leaf) of the yellow-green leaf mutant yg/2 and its wild-type GL11, and 183 recessive individuals (yellow-green leat) isolated from F3 were identified. Specifically, by a CTAB method, genomic DNAs of seedling leaves of the parents and the population were extracted, and PCR amplification was conducted using primer pairs in Table 3. A reaction system included: 1 pL to 2 pL of a DNA template (80 ng to 150 ng), 0.6 uL of each of the upstream primer and the downstream primer, 2 pL of a 10xEasy Taq Buffer (Mg? free), 2 uL of dNTPs (2.5 mM), 0.2 uL of Easy Taq Polymerase (TransGen, AP111), and adding ddH>O to make up to 20 uL. A reaction procedure of the PCR amplification included: initial denaturation at 95°C for 5 min; denaturation at 95°C for 30 sec, annealing at (Tm-5)°C for 30 sec, and extension at 72°C (1 min/kb), conducting 35 cycles; and extension at 72°C for 8 min. PCR amplification products were identified by 0.8% to 2% agarose gel, and after electrophoresis for about 30 min, the products were observed in a gel imaging system and photographed for storage. The results were shown in FIG. 11, namely photographs of the development and identification of InDel markers for 45 individuals out of 338 samples. It was seen from FIG. 11 that the genotypes of the identified 45 samples with yellow-green leaves were all the same, which were consistent with the yellow-green leaf mutant yg/2.

[81] The developed InDel markers were used to identify genotypes of domestic and foreign planting resources, and an identification process was the same as above, as shown in Table 4 and FIG. 12.

[82] Table 4 40 soybean varieties with known leaf color at home and abroad

[83]

No. Variety name Known leaf color Molecular marker identification results 1 vel2 Yellow-green Yellow-green 2 GL11 Green Green 3 Jack Green Green 4 Williams 82 Green Green 5 Yangyandou Green Green 6 Changnong 28 Green Green 7 Changnong 29 Green Green

8 Kenfeng 18 Green Green 9 Kenfeng 19 Green Green

Heinong 33 Green Green 11 Heinong 61 Green Green 12 Dongnong 48 Green Green 13 Dongnong 49 Green Green 14 Dongnong 50 Green Green

Qihuang 34 Green Green 16 Dengke 4 Green Green 17 Dengke 13 Green Green 18 Heihe 18 Green Green 19 Heihe 38 Green Green

Heihe 42 Green Green 21 Heihe 43 Green Green 22 Henong 64 Green Green 23 Henong 65 Green Green 24 Suinong 39 Green Green

Zhonghuang 38 Green Green

26 Zhonghuang 65 Green Green 27 Zhonghuang 73 Green Green 28 Heinong 33 Green Green 29 Heinong 39 Green Green 30 Heinong 41 Green Green 31 Heinong 51 Green Green 32 Zhonghuang 13 Green Green 33 Zhongji 602 Green Green 34 Zhongpin 661 Green Green 35 Kenfeng 18 Green Green 36 Kenfeng 19 Green Green 37 Kenfeng 20 Green Green 38 Changnong 18 Green Green 39 Changnong 19 Green Green 40 Changnong 20 Green Green

[84] According to Table 4 and FIG. 12, it was seen that 39 green leaf genotypes were the same as those of the wild-type GL 11. The identification results of the InDel marker were consistent with the known leat colors of soybean varieties, indicating that the InDel marker was successfully developed.

[85] Examples

[86] Detection and use of editing efficiency of gene Gmvg/2 in soybean

[87] A DNA of soybean GLI11 leaves was extracted, a full length of the gene

Gmygl2 was amplified, and ligated to a plant editing vector pCEPO1 (with a resistance in £. coli of kanamycin resistance, and a resistance in plants of glufosinate-ammonium resistance), to obtain a Gmyg/2-edited vector.

[88] The wild-type GL11 mature soybean seeds with smooth surface, no damage, no disease spots and no cracks were selected and sterilized by chlorine gas for 14 h.

The sterilized seeds were ventilated on an ultra-clean table to completely volatilize the chlorine gas, and then germinated in a germination medium for 6 h. 1/2 hypocotyl of soybean was removed, the soybean was cut longitudinally along the hypocotyl, and the remaining hypocotyl was used as a recipient material for Agrobacterium-mediated transformation.

[89] By Agrobacterium-mediated transformation of soybean cotyledon nodes, a

Gmygl2-knockout gene was introduced into Jack, to obtain a Gmyg/2 mutant explant containing an editing vector of the regulatory gene Gmyg/2; the explant was cultured in a co-culture medium (BS powder: 0.32 gL, MES: 3.9 g L*t, sucrose: 30 g Ll, agar: 5 gL! AS: 40 mg Lt, B5 organic: 112 mg-L™) in the dark at 22°C for 5 d; the explant was cultured in an SI-I medium under strong light for 7d; large shoots of the explant (wild-type GL11) were excised, and cultured in an SI-IT medium under strong light for 14 d; cotyledons and hypocotyls of the explant was excised, and subcultured every 14 din an SE medium; about 3 cm of clump buds were cut off and put into a rooting medium for rooting, the plants with well-developed roots in an RM rooting medium were transferred to soil for planting. Through a Bar resistance test, 30 resistant plants were screened. After 5 months of cultivation in the greenhouse, the pods began to mature, and harvest was completed after 6 months.

[90] The harvested T1 generation seeds were planted in a greenhouse, and it was observed that after the gene Gmyg/2 was knocked out and transformed into Jack, the leaf color of transgenic plants (gene-edited seedling YGL2-SP1) was the same as that of the mutant vg/2 (FIG. 13), and encoded protein were shown in FIG. 15. The results fully demonstrated that the gene Gmyg!2 and its encoded protein were the functional gene and protein that led to the phenotype of yellow-green leaf, which could be used to screen yellow-green leaf genes and provide uses thereof.

[91] Although the above embodiments have described the present disclosure in a thorough manner, it is only some but not all embodiments of the present disclosure,

and other embodiments may be obtained without inventive step according to the present embodiments, all of which fall within the scope of protection the present disclosure.

Sequence Listing Information:

DTD Version: V1 3

File Name: HKJP202112815-sequence listing. xml

Software Name: WIPO Sequence

Software Version: 2.1.2

Production Date: 2022-08-16

General Information:

Current application / Applicant file reference: HKJP202112815

Applicant name: Institute of Crop Sciences, Chinese Academy of Agricultural

Sciences

Applicant name / Language: en

Invention title: GENE Gmygl2 RELATED TO SOYBEAN PLANT HEIGHT

AND LEAF COLOR, INSERTION-DELETION (InDel) MARKER OF GENE

Gmygl2, AND USE THEREOF ( en)

Sequence Total Quantity: 10

Sequences:

Sequence Number (ID): 1

Length: 7475

Molecule Type: DNA

Features Location/Qualifiers: - source, 1..7475 > mol type, other DNA > note, DNA sequence of the gene Gmygl2 > organism, synthetic construct

Residues: aacagagtaa titgctgagc acgtgtgget tgcttggacg agagtgastt gcagtgcata 60 gtgattgaga gcagaagaag ctgcttaagc ttaacttaaa tgggttttag aattagcggt 120 tectcctctt catcatgctt ttgtttgcaa cgtacgcttc ttcctccgtt aacagcaata 180 gccacttcca ctcgtggctt caagaggagg agtaattgca taccgagttg ttcagtatct 240 tatcgcaagt ttgttgagtt tgctttggat gaaaccacac tccacactca cttgatccct 300 tcgcctttac aggtttetct tattccettte tttaattatt gccatatcct tgtgtgtgtt 360 ggacctgtaa tggttgatta gggttagtta gttccactct gttttagaac ttttgtcgtt 420 tgttgacact gaaccctaat ccaaagtgaa ttttccgctt tgtasttttt taaaacttaa 480 tttgcagata tcttcaagcc aatttaggca gtgtttggta tatgtgtcag ctgaaactga 540 caaaacataa ttatgtgaat tctcctttta tgttgcattt tagaaataga acaggactta 600 aagttgaaac aaaggcccat tggggttcta tagaatgtca atgtgtgaaa tcaaggacat 660 ctgaacgaaa tgcaacattt cattctgttt tgtctatata ccaaatgcta ccttacctta 720 ctgatgcatc tgtacaaaac cgaagcaaat tttgataaaa agaatttttt attttatttt 780 aaatgagaga gttatatgtt aggccttttt atttgtaggc ctaaaacttg aactttagta 840 ggcttaccct ttggccgacc ctgaggaaga ggagaaaaag aaattgaatg aactgctcta 900 taaaacaagg tctctgggtt agttctcaaa accaaaagat tgttaccata ctttggatgg 960 cagtgtttaa taaacataaa tgaagtatca ctttggtttg gttaatgttt aataaacatc 1020 ttccagtgaa tttggaagaa aaaaatgaga tgaaaaagag ttttactatg actatgtgtc 1080 atttcatgtc ctcatgtttt aaatttttgt gttgccctgt tgaggatgtg gstgtactgtt 1140 tcaatcacca agtcttggct tctcagaata gaactagctc ttcaattaat aagccattaa 1200 ctcttttgtt tcttcttggt attgagttat tcaactttca atttcattta tattttgtta 1260 tggtattatt ttaaatttag ttaaattgaa cacataattt ctttgcagat tcttaattgt 1320 gattttttgt ccctagttgt tctccactcc tctgctttgt ttgcttaatt tgctctccac 1380 cattatttgt gtgcatagat cttcaatcta tactttttta tgttstcttg ctaggaaaag 1440 tacaatttca tgaattccaa ggatggtaaa ggaactctta gtatgctatc atttgaaggt 1500 gccaaaatta ggcttctacg aagtttgatc attgagacag aaacaatgca ggtetttttt 1560 ttaatgccta tcaattgtca aattgatata tggttggtat ctactactcc ctttgtccca 1620 taatggctga tgtttaaggt tttttcacac agaccaagaa agcaagtatt ccaagagaat 1680 taacatttag taggaaacaa ttttactaaa atatccttac tctctctctt aatttcaata 1740 aatgtactat taagtcttcc aagacaacat tatttattac aagacaaaat tggaatagat 1800 gttttaatat ctcattgaaa agtgagaaca tcagtcaatt tggaattttt tcaaaggtta 1860 aaactgcagt cattttggga catgtcttaa caaaatatgc cttttgtatg gtaccttgta 1920 atttattata gccataacaa tttcaaaatt tgactttgtc tgtatttatt tctgccagtc 1980 tcagttttct ttttctttta ccaacaaagg atccttcatc taaaacaatg catgttatga 2040 tgttaacaga attagtaagc atcctgttgt atctgtttca atatgcaatc tctgatcccc 2100 atgaggtata acacttgttg gaatatctca taacatccat attatatctc agaatatttt 2160 agagtatctt agaaattctc ttaggattag ctaatgtgat ttctcagttt ccattttect 2220 aatgttttat sttttgcaat tccatttcaa caacacatgc aaaaaattga catgtcattg 2280 ctgaagtgtt ctgctgtctt ttgtgacctg tctgtggaaa ggttgtttta caatatgctt 2340 actgcctctt tttttatgtc ttcaacttct tatgcaggtt ttggatttta ctgtctttce 2400 aaaagcagaa tatgacatac ccatattttg tgctaacttt ttcacctctg ctaaaacaaa 2460 cattgttgtg ttgtaagtta gcatcttttt ccactgttgc tatccactta tataaatgta 2520 taaaagatgt tcttatattt aactcttagt tttggaaaag tctagcaatg tgttctatgt 2580 ttgcaatttt taaacttatt cctaaactgc atttcagcta tggaaattta tgtatgacaa 2640 tgcaatattt tcccatccaa aaagaattca ccactaatta tgaaaaccta tcttgggttg 2700 ctttgcaaat cgaagtggga atcattcagt tttaaattaa aaaataagcc agcctctgat 2760 ctatgtggat atatactttc aagaggtaca cacaacatag aacttttatg tcttcatttg 2820 getgtggtaa atgcatatgt ttcattgcct agtaatacac accaaagtag gtgtatcaag 2880 tatcaaccag cctcagcagc aggtatttga ttgtacttgg taaaagaaaa taccatattt 2940 cattagttgt aaatttactt gcataatcac ttgaaaagga tgtttcattc ttttgtaaaa 3000 tgggttcatg tctcttttat ggtatattcc cgattgaata aaagcatgca tttccagaca 3060 agaagattga atgtggagga attgtccaca tgatgaacca aaccgagctg tgaagggcag 3120 agtgaagtta ccattttagg agtagatgtc ctagatctca atgttagttt aagcgagaaa 3180 atagtagttg ttttactcca gccctttggt tttgctattt tttttctatg aacctttaaa 3240 aagatgaagt ttagccctaa tcagcataat aaagatcaca gttctttact ttcagagttg 3300 cagtgcttat tttattctct ttagaatgtg gtcttgtaaa taaaggaccc taaaacctta 3360 gttcagtaaa ttttgctata tttgtcttct taagatatca tgtcatattc tatgtaaagg 3420 agattttagg attgaaagca tctttgattg actatttgaa attatgttat atataccaaa 3480 ttatccttgt tatatccata ttagcgtttg cttttcattt tcttatgtat tatagcttta 3540 caataaaaaa aattagatct gaagttgatg tatactttaa tttgagttta taggcttttg 3600 gatgattggt attgactctt catccaatct tagaaagaac tatttggtga agctgttata 3660 aacttataat aaatgagaaa aaaaatgcct ttctttatca gtaaatagaa actcaaggat 3720 ttaatgtgtg ttgcatgtct actaaaattt gccacctact cagctgtgga agaatattat 3780 agtgtcatcc ttttagataa tctactgtta gctttcttag gtcgcattac ttctccctca 3840 atttaattgt gtttagtcat attgtcagtg catgtgcaaa tttaatggta cttggstgtt 3900 tatacttgtc caaaattaac ataatggaaa agaccaagct ttggcaagaa ataaagccta 3960 aagcaaatcc atgaaaaatc tgtgacagtc tgtagtgctc atttctcccc aatattgttt 4020 acaccttaaa cgtcttggaa atctaatatt tgttcaccga catcactggt aatgtgataa 4080 agtttttcce cactgtgcta tttatagctt caaaattatt taagaacaaa ttactgtatg 4140 taatgcagag cactaacctt gaattaattg tgccagggac cttaacccct tgcatgatat 4200 catcaatcag catgagtaca aggagaagta ctttaaaagc ttaattcctc tcggccttaa 4260 atatgctgag gtaactgcaa atattttagg gaaaattata aggattattg gtagaagtat 4320 attttagaaa aagttaatca ctcgactgca taacaatgtg aaaagttttg ttaataacag 4380 ctacttattg acctaattag ccacatgagt gatgcattgt tctaaaactg gatttacttt 4440 taatatgaat tctatatttt ttacttttcc ttttagtatt gattcattga agtcctcatg 4500 gatgcatctc cgtttttctg taaattatag agattctttc ttaaaggacc tagtgagttg 4560 acttaacaca gactttatgc agctttttcc atggggaggg aagctcacaa gtgagtccat 4620 aaattttttt tcaccaattg tcatctggac aaagtttacc tcaaacccag aaaaatatga 4680 tattctgtat tccgcattta gggaatatta caaggtaata gccccagcat catgatcaac 4740 tttatgaatg acaattcata ttttgctaaa tactctctcc gttcctatat ataagacctt 4800 tgaactaatt cacactcttt aataaaattg gttaatatag ttagtgacat taaatttgtc 4860 aataatttat atttttttat aattttcctt aatgttttta aaattaatca tctctccctt 4920 tccacctaat ttcttttcge ctaattaatt aatgtttggg gatggaataa tgattatgta 4980 atttgtggct actcatcttc tccaatccta ataagagagt ttatctcatt aatttccagc 5040 atttattttt cttggctaaa aagtttgtta attaattaag ggtattttag taaaaaaaat 5100 aatacaagag acaattagaa aagggtctta taaaaaggga caagaaattt tctgaaaatg 5160 gtcttatata taaggatgga gggagtatta atacatttag tgaactttaa taaagttgac 5220 cttgtacatg tgtaattttg ttatcttgat gtctgttacc ctggcaggta tggttgaaat 5280 tgatatgcaa agcagataaa gagacagatg aatctcagat tttccacaac ctcgaagcac 5340 aacatagata tctaacatgg agagttgaaa aggtagttca catttgctaa aattttett 5400 gacagccaat cataaatatg attggtgatc tagccataag caaacttata tttgtcccag 5460 gtataattgg tttaggattt tctaattgat caaggatctt cactcattca gctagcatcg 5520 gttgaagata atttagatga taatattaac aaagatggtt ctgtttgaat atgatgtctt 5580 caaaaaacca acctaacttc agttattttc tgcattcaaa tctcaacaat aaaattggca 5640 aataagtttg aaatgaagtt ttgaacttca aactatataa acaagggaca gccaatccta 5700 tcaggacttt aatttagaat gtatagggcc tgtttggata actctctaaa aaccactcta 5760 ggagagaaat agaaggaaaa atgaaaaagc ttctccataa tctatttcta attcagtttt 5820 aatggacctg gttcatggac atacttgaaa gtaggatcca ggacaagstg ttttgaagaa 5880 gctgattggt gacacacttg ccaaggtagg ccaaatttgt gctcgattta tttaattcct 5940 ctacatccta gttccacaga tcctcacttc ctgttttata ctaggatatg ctgagaagct 6000 ttctctttaa tggagttgat gaactaggaa gcaaaacatt caatgattat tttccacgct 6060 actgctgtca agagggaact ctaaataaaa aaggcaatgt tattgggaag tcctttgaaa 6120 atcgcccatg gaatgctaga ggagaattta ttggtaacag attttcaaat tggcctgtct 6180 aattgctgct ttcttttcat ctctgcttgg cgtgtttttt atcctgctca tctgttatca 6240 ttctgtttat gtcctgcggc cttctattag ggtcttacaa gtaaaaaact atgctgtgat 6300 caatggattc tttcacctgc cttttcttct tttatattaa aattggtcct gcatatttat 6360 ctatggcatt ttgcatgtat atgttctttg attacttgaa gttgcttaac tggcagtttg 6420 atttgagatg cgagagacat tgttgaagcc agtaggactt gaattctatt attcatgatt 6480 aagaatgaga tgatttcagt aatgaaatag ttgtatcgtt tagcaagcta gcaaatattt 6540 ttgtagaagc aagcacttac aagcatacta tatgtagcac aagaaatctt cagaattata 6600 atgatatgtt ggggegtect tgaaaaaaaa aattggactt gtgagaaagt agttcagaac 6660 ttttgtaagt tttttttttc aatttatttt aacaagattt acagagaaac tggttgaagt 6720 aagttttttt ttaacctatt tcagtaattc agttaatttt atgaacaaat ttttatttgg 6780 taggagttta ttaaataast ggttaatgaa attctttatt taaatgcatc cttagtttat 6840 agtaattttt aaggatcctt cagctggatt ggcgttgatg tgtagcttte aggccaaggt 6900 atacttaccg cggtgctgac atttgatctt tagaatattt taggtgaatg actgtacact 6960 ggagaagctt gtgcacacgt ttgcagtcaa acttttacaa ggcgtgccac tggttatatt 7020 ttcggctata gaggttctgc aagcatacaa aagaccgctg taatttttgt agacaagtta 7080 tgccataaga tgcttgtaat atattatgga aagaattttc tgtaacaaat ctcaggctca 7140 gcacctatgg ctctacatgt ttgtaatatt aacattcgta ttgtatcagt caaaaaaaca 7200 ttcgtattgg aagattctag ttatacatta actgcagccg tgtccgatta gcttcaagtc 7260 atagtagaaa tttaagtgta atattttacc ttctgtaaaa taaagtttaa tgttttacct 7320 tttgtaaaaa aaaatgtaat attttagcat gcagtttaga ttaatttgta taacccaaaa 7380 attgtcactc gatttaattt ttgggttaaa tcaagatttt ttttaatacg attgggctct 7440 ttatttggca tgctggctaa attgggtctc tttgt 7475

Sequence Number (ID): 2

Length: 990

Molecule Type: DNA

Features Location/Qualifiers: - source, 1.990 > mol type, other DNA > note, CDS sequence of the gene Gmygl2 > organism, synthetic construct

Residues: atgogtttta gaattagcgg ttcctcctct tcatcatgct tttgtttgca acgtacgctt 60 cttcctccgt taacagcaat agccacttcc actcgtggct tcaagaggag gagtaattgc 120 ataccgagtt gttcagtatc ttatcgcaag tttsttgast ttgctttgga tgaaaccaca 180 ctccacactc acttgatccc ttcgccttta caggaaaagt acaatttcat gaattccaag 240 gatggtaaag gaactcttag tatgctatca tttgaaggtg ccaaaattag gcttctacga 300 agtttgatca ttgagacaga aacaatgcag sttttggatt ttactgtctt tccaaaagca 360 gaatatgaca tacccatatt ttgtgctaac tttttcacct ctgctaaaac aaacattgtt 420 gtgttggacc ttaacccctt gcatgatatc atcaatcagc atgagtacaa ggagaagtac 480 tttaaaagct taattcctct cggccttaaa tatgctgagc tttttccatg gggagggaag 540 ctcacaagtg agtccataaa ttttttttca ccaattgtca tctggacaaa gtttacctca 600 aacccagaaa aatatgatat tctgtattcc gcatttaggg aatattacaa ggtatggtte 660 aaattgatat gcaaagcaga taaagagaca gatgaatctc agattttcca caacctcgaa 720 gcacaacata gatatctaac atggagagtt gaaaaggatc caggacaagg tgttttgaag 780 aagctgattg gtgacacact tgccaaggat atgctgagaa gctttctctt taatggastt 840 gatgaactag gaagcaaaac attcaatgat tattttccac gctactgctg tcaagaggga 900 actctaaata aaaaaggcaa tgttattggg aagtcctttg aaaatcgccc atggaatgct 960 agaggagaat ttattgaata ttttaggtga

Sequence Number (ID): 3

Length: 329

Molecule Type: AA

Features Location/Qualifiers: - source, 1..329 >mol type, protein > note, Amino acid sequence of the protein encoded by the CDS sequence > organism, synthetic construct

Residues:

MGFRISGSSS SSCFCLQRTL LPPLTAIATS TRGFKRRSNC

IPSCSVSYRK FVEFALDETT 60

LHTHLIPSPL QEKYNFMNSK DGKGTLSMLS FEGAKIRLLR

SLIETETMQ VLDFTVFPKA 120

EYDIPIFCAN FFTSAKTNIV VLDLNPLHDI INQHEYKEKY

FKSLIPLGLK YAELFPWGGK 180

LTSESINFFS PIVIWTKFTS NPEKYDILYS AFREYYKVWL

KLICKADKET DESQIFHNLE 240

AQHRYLTWRV EKDPGQGVLK KLIGDTLAKD MLRSFLFNGV

DELGSKTFND YFPRYCCQEG 300

TLNKKGNVIG KSFENRPWNA RGEFIEYFR

329

Sequence Number (ID): 4

Length: 980

Molecule Type: DNA

Features Location/Qualifiers: - source, 1..980 > mol type, other DNA > note, Gmygl2 sequence of the ygl2 mutant > organism, synthetic construct

Residues: atgggtttta gaattagcgg ttcctcctct tcatcatgct tttgtttgca acgtacgctt 60 cttcctccgt taacagcaat agccacttcc actcgtggct tcaagaggag gagtaattgc 120 ataccgagtt gttcagtatc ttatcgcaag tttgttgast ttgctttgga tgaaaccaca 180 ctccacactc acttgatccc ttcgccttta caggaaaagt acaatttcat gaattccaag 240 gatggtaaag gaactcttaa tttgaaggtg ccaaaattag gcttctacga agtttgatca 300 ttgagacaga aacaatgcag gttttggatt ttactgtctt tccaaaagca gaatatgaca 360 tacccatatt ttgtgctaac tttttcacct ctgctaaaac aaacattgtt gtgttggacc 420 ttaacccctt gcatgatatc atcaatcagc atgagtacaa ggagaagtac tttaaaagct 480 taattcctct cggccttaaa tatgctgagc tttttccatg gggagggaag ctcacaagtg 540 agtccataaa ttttttttca ccaattgtca tctggacaaa gtttacctca aacccagaaa 600 aatatgatat tctgtattcc gcatttaggg aatattacaa ggtatggttg aaattgatat 660 gcaaagcaga taaagagaca gatgaatctc agattttcca caacctcgaa gcacaacata 720 gatatctaac atggagagtt gaaaaggatc caggacaagg tgtittgaag aagctgattg 780 gtgacacact tgccaaggat atgctgagaa gctttctctt taatggagtt gatgaactag 840 gaagcaaaac attcaatgat tattttccac gctactgctg tcaagaggga actctaaata 900 aaaaaggcaa tgttattggg aagtcctttg aaaatcgccc atggaatgct agaggagaat 960 ttattgaata ttttaggtga 980

Sequence Number (ID): 5

Length: 98

Molecule Type: AA

Features Location/Qualifiers: - source, 1..98 > mol type, protein > note, Amino acid sequence of the protein encoded by Gmygl2 from the ygl2 mutant > organism, synthetic construct

Residues:

MGFRISGSSS SSCFCLQRTL LPPLTAIATS TRGFKRRSNC

IPSCSVSYRK FVEFALDETT 60

LHTHLIPSPL QEKYNFMNSK DGKGTLNLKV PKLGFYEV

98

Sequence Number (ID): 6 13 Length: 22

Molecule Type: DNA

Features Location/Qualifiers: - source, 1.22

> mol type, other DNA > note, Forward primer for ID 105 > organism, synthetic construct

Residues: aagagtttta ctatgactat gt 22

Sequence Number (ID): 7

Length: 22

Molecule Type: DNA

Features Location/Qualifiers: - source, 1.22 > mol type, other DNA > note, Reverse primer for ID 105 > organism, synthetic construct

Residues: ataaatacag acaaagtcaa at 22

Sequence Number (ID): 8

Length: 21

Molecule Type: DNA 3 Features Location/Qualifiers: - source, 1..21 > mol type, other DNA > note, Forward primer for PCR amplication of RNA from GL 11 > organism, synthetic construct

Residues: atgggtttta gaattagcgg t 21

Sequence Number (ID): 9 13 Length: 26

Molecule Type: DNA

Features Location/Qualifiers: - source, 1.26

> mol type, other DNA > note, Reverse primer for PCR amplication of RNA from GL 11 > organism, synthetic construct

Residues: cctaaaatat tcaataaatt cteete 26

Sequence Number (ID): 10

Length: 63

Molecule Type: AA

Features Location/Qualifiers: - source, 1..63 > mol type, protein > note, Amino acid sequence of the protein encoded by the gene Gmygl2 from YGL2-SPI > organism, synthetic construct

Residues:

MGFRISGSSS SSCFCLQRTL LPPLTAIATS IVASRGGVIA

YRVVQYLASL LSLLWMKPHS 60

TLT

63

END i Alum. overslon=T1.0" encoding=TUTF-Qn Tx 2 <!DOCTYPE ST26SequenceListing PUBLIC "-//WIPO//DTD Sequence Listing 1.3//EN" "ST265equenceListing V1 3.dtd"> 3 <3T268equencelisting drdversion="NVL 3% filoName="HRJIP20Z1128153-sagquances listing. sal? softwaraNanme= WIPO Seguange” softwareVersion="2 i.e oroductionDats="2022-08-18%> & <ApplicantFileReference>HKJP202112815-/ApplicantFileReference> <AppiicantNeme langusselcdse=Men">Institute of Crop Sciences, Chinese Academy of

Agricultural Sciencesd</ApplicantName> & <InpvantlionTitle lanymuiagelods="en">GENE Gmygl2 RELATED TO SOYBEAN PLANT HEIGHT

AND LEAF COLOR, INSERTION-DELETION (InDel) MARKER OF GENE Gmygl2, AND USE

THEREOF </InventionTitle: i <SeqguencaTotaluantity>10</SeguenceTotalQuantity> 5 <SequenceData sequenceIDNumber="iN>

G <INSDSeqd»>

LO <INSDSeq length>7475</IN3DSeqg length» 11 <INSDSeq moltype>DNA</INSDSeg moltype> 12 <INSDSeq divislion»PAT</INSDSeqg division» 12 <IN3D3eq feature-table> 14 <INSDFeature>

Ls <IN3DFeature key>source</IN3DFeature key> 18 <IN3DFeature location>»l..7475</INSDFeature location»

Lj <INSDFsature qualsg> is <INSDuuelifier> 1% <INSDoualifier name>mol type“ /INSDQualifier name> a <INSDQualifisr value>other DNA</INSDQualifier valuer a </INSDOualifier> 22 <INSDOualifier id="g2"> 23 <IN3DQualifier name>note</INSDQualifier name> 24 <INSDQualifiler value>DNA sequence of the gene

Gmygl2/INSSQualifier value» zE «</INSDOQualifier»> es <INSDQualifler id="gl®> 7 “INSDoualifier name>organism</INSDQualifier name> 28 <IN3DQualifier value>synthetic construct</INSDQualifier value» 23 </INSDOQuali fier </INSDFsature guals> 3 </INSDFeature> 32 </INSDSeqg feature-tabhlex <INSDSeq sequencevaacagagtaatttgctgagcacgtgtggcttgcttggacgagagtgagttgcagtg catagtgattgagagcagaagaagctgcttaagcttaacttaaatgggttttagaattageggttectectett catcatgettttgtttgcaacgtacgettettcctcegttaacagcaatagccacttccactecgtggcttcaag aggaggagtaattgcataccgagttgttcagtatcttatcgcaagtttgttgagtttgctttggatgaaaccac actccacactcacttgatccettegcctttacaggtttectcttattcctttetttaattattgccatatccttg tgtgtgttggacctgtaatggttgattagggttagttagttccactetgttttagaacttttgtecgtttgttga cactgaaccctaatccaaagtgaattttcegctttgtagttttttaaaacttaatttgcagatatcttcaagcc aatttaggcagtgtttggtatatgtgtcagctgaaactgacaaaacataattatgtgaattctccttttatgtt gcattttagaaatagaacaggacttaaagttgaaacaaaggcccattggggttctatagaatgtcaatgtgtga aatcaaggacatctgaacgaaatgcaacatttcattectgttttgtctatataccaaatgctaccttaccttact gatgcatctgtacaaaaccgaagcaaattttgataaaaagaattttttattttattttaaatgagagagttata tgttaggcetttttatttgtaggcctaaaacttgaactttagtaggettaccctttggccgaccctgaggaaga ggagaaaaagaaattgaatgaactgctctataaaacaaggtctctgggttagttctcaaaaccaaaagattgtt accatactttggatggcagtgtttaataaacataaatgaagtatcactttggtttggttaatgtttaataaaca tcttccagtgaatttggaagaaaaaaatgagatgaaaaagagttttactatgactatgtgtcatttcatgtect catgttttaaatttttgtgttgccctgttgaggatgtggtgtactgtttcaatcaccaagtecttggcttctcag aatagaactagctcttcaattaataagccattaactecttttgtttettettggtattgagttattcaactttca atttcatttatattttgttatggtattattttaaatttagttaaattgaacacataatttctttgcagattectt aattgtgattttttgtccctagttgtteteccactcctectgetttgtttgettaatttgctctccaccattattt gtgtgcatagatcttcaatctatacttttttatgttgtettgctaggaaaagtacaatttcatgaattccaagg atggtaaaggaactcttagtatgctatcatttgaaggtgccaaaattaggcttctacgaagtttgatcattgag acagaaacaatgcaggtgttttttttaatgcctatcaattgtcaaattgatatatggttggtatctactactce ctttgtcccataatggctgatgtttaaggttttttcacacagaccaagaaagcaagtattccaagagaattaac atttagtaggaaacaattttactaaaatatccttactctectctcettaatttcaataaatgtactattaagtctt ccaagacaacattatttattacaagacaaaattggaatagatgttttaatatctcattgaaaagtgagaacatc agtcaatttggaattttttcaaaggttaaaactgcagtcattttgggacatgtcttaacaaaatatgccttttg tatggtaccttgtaatttattatagccataacaatttcaaaatttgactttgtetgtatttatttctgccagtc tcagttttetttttettttaccaacaaaggatccttcatctaaaacaatgcatgttatgatgttaacagaatta gtaagcatcctgttgtatctgtttcaatatgcaatctctgatccccatgaggtataacacttgttggaatatct cataacatccatattatatctcagaatattttagagtatcttagaaattctcttaggattagctaatgtgattt ctcagtttccattttcctaatgttttatgttttgcaattccatttcaacaacacatgcaaaaaattgacatgtec attgctgaagtgttetgetgtcttttgtgacctgtctgtggaaaggttgttttacaatatgcttactgcctett tttttatgtettcaacttcttatgcaggttttggattttactgtetttccaaaagcagaatatgacatacccat attttgtgctaactttttcacctctgctaaaacaaacattgttgtgttgtaagttagcatctttttccactgtt gctatccacttatataaatgtataaaagatgttcttatatttaactcttagttttggaaaagtctagcaatgtg ttctatgtttgcaatttttaaacttattcctaaactgcatttcagctatggaaatttatgtatgacaatgcaat attttcccatccaaaaagaattcaccactaattatgaaaacctatcttgggttgetttgcaaatcgaagtggga atcattcagttttaaattaaaaaataagccagcctctgatctatgtggatatatactttcaagaggtacacaca acatagaacttttatgtecttcatttgggtgtggtaaatgcatatgtttcattgcctagtaatacacaccaaagt aggtgtatcaagtatcaaccagcctcagcagcaggtatttgattgtacttggtaaaagaaaataccatatttca ttagttgtaaatttacttgcataatcacttgaaaaggatgtttcattettttgtaaaatgggttcatgtetett ttatggtatattcccgattgaataaaagcatgcatttccagacaagaagattgaatgtggaggaattgtccaca tgatgaaccaaaccgagctgtgaagggcagagtgaagttaccattttaggagtagatgtcctagatctcaatgt tagtttaagcgagaaaatagtagttgttttactccagccctttggttttgetattttttttctatgaaccttta aaaagatgaagtttagccctaatcagcataataaagatcacagttctttactttcagagttgcagtgettattt tattctectttagaatgtggtcttgtaaataaaggaccctaaaaccttagttcagtaaattttgctatatttgtec ttcttaagatatcatgtcatattctatgtaaaggagattttaggattgaaagcatctttgattgactatttgaa attatgttatatataccaaattatccttgttatatccatattagecgtttgettttcattttcttatgtattata gctttacaataaaaaaaattagatctgaagttgatgtatactttaatttgagtttataggettttggatgattg gtattgactcttcatccaatcttagaaagaactatttggtgaagctgttataaacttataataaatgagaaaaa aaatgcctttectttatcagtaaatagaaactcaaggatttaatgtgtgttgcatgtctactaaaatttgccacc tactcagctgtggaagaatattatagtgtcatcecttttagataatctactgttagectttcttaggtegcattac ttcetccctcaatttaattgtgtttagtcatattgtcagtgcatgtgcaaatttaatggtacttgggtgtttata cttgtccaaaattaacataatggaaaagaccaagctttggcaagaaataaagcctaaagcaaatccatgaaaaa tctgtgacagtctgtagtgctcatttctccccaatattgtttacaccttaaacgtcttggaaatctaatatttg ttcaccgacatcactggtaatgtgataaagtttttccccactgtgctatttatagcttcaaaattatttaagaa caaattactgtatgtaatgcagagcactaaccttgaattaattgtgccagggaccttaaccccttgcatgatat catcaatcagcatgagtacaaggagaagtactttaaaagcttaattccteteggccttaaatatgctgaggtaa ctgcaaatattttagggaaaattataaggattattggtagaagtatattttagaaaaagttaatcactcgactg cataacaatgtgaaaagttttgttaataacagctacttattgacctaattagccacatgagtgatgcattgtte taaaactggatttacttttaatatgaattctatattttttacttttecttttagtattgattcattgaagtcct catggatgcatctcegtttttctgtaaattatagagattctttcttaaaggacctagtgagttgacttaacaca gactttatgcagctttttccatggggagggaagctcacaagtgagtccataaattttttttcaccaattgtcat ctggacaaagtttacctcaaacccagaaaaatatgatattctgtattccgcatttagggaatattacaaggtaa tagccccagcatcatgatcaactttatgaatgacaattcatattttgctaaatactectctceegttcctatatat aagacctttgaactaattcacactctttaataaaattggttaatatagttagtgacattaaatttgtcaataat ttatatttttttataattttccttaatgtttttaaaattaatcatctetccctttccacctaatttettttegc ctaattaattaatgtttggggatggaataatgattatgtaatttgtggctactcatcttctccaatcctaataa gagagtttatctcattaatttccagcatttatttttcttggctaaaaagtttgttaattaattaagggtatttt agtaaaaaaaataatacaagagacaattagaaaagggtcttataaaaagggacaagaaattttctgaaaatggt cttatatataaggatggagggagtattaatacatttagtgaactttaataaagttgaccttgtacatgtgtaat tttgttatettgatgtctgttaccctggcaggtatggttgaaattgatatgcaaagcagataaagagacagatg aatctcagattttccacaacctcgaagcacaacatagatatctaacatggagagttgaaaaggtagttcacatt tgctaaaattttctttgacagccaatcataaatatgattggtgatctagccataagcaaacttatatttgtccc aggtataattggtttaggattttctaattgatcaaggatcttcactcattcagctagcatcggttgaagataat ttagatgataatattaacaaagatggttctgtttgaatatgatgtcttcaaaaaaccaacctaacttcagttat tttctgcattcaaatctcaacaataaaattggcaaataagtttgaaatgaagttttgaacttcaaactatataa acaagggacagccaatcctatcaggactttaatttagaatgtatagggcctgtttggataactctctaaaaacc actctaggagagaaatagaaggaaaaatgaaaaagcttctccataatctatttctaattcagttttaatggacc tggttcatggacatacttgaaagtaggatccaggacaaggtgttttgaagaagctgattggtgacacacttgcc aaggtaggccaaatttgtgctcgatttatttaattcectctacatcctagttccacagatcctcacttectgttt tatactaggatatgctgagaagctttctctttaatggagttgatgaactaggaagcaaaacattcaatgattat tttccacgctactgctgtcaagagggaactctaaataaaaaaggcaatgttattgggaagtcctttgaaaatcg cccatggaatgctagaggagaatttattggtaacagattttcaaattggcctgtctaattgectgectttetttte atctetgettggegtgttttttatcctgctcatctgttatcattctgtttatgtectgeggccttctattaggg tcttacaagtaaaaaactatgctgtgatcaatggattectttcacctgccttttettettttatattaaaattgg tcctgcatatttatctatggcattttgcatgtatatgttctttgattacttgaagttgecttaactggcagtttg atttgagatgcgagagacattgttgaagccagtaggacttgaattctattattcatgattaagaatgagatgat ttcagtaatgaaatagttgtatcgtttagcaagctagcaaatatttttgtagaagcaagcacttacaagcatac tatatgtagcacaagaaatcttcagaattataatgatatgttggggggtgcttgaaaaaaaaaattggacttgt gagaaagtagttcagaacttttgtaagttttttttttcaatttattttaacaagatttacagagaaactggttg aagtaagtttttttttaacctatttcagtaattcagttaattttatgaacaaatttttatttggtaggagttta ttaaataagtggttaatgaaattctttatttaaatgcatccttagtttatagtaatttttaaggatccttcagc tggattggegttgatgtgtagctttcaggccaaggtatacttaccgeggtgctgacatttgatctttagaatat tttaggtgaatgactgtacactggagaagcttgtgcacacgtttgcagtcaaacttttacaaggcgtgccactg gttatatttteggctatagaggttctgcaagcatacaaaagaccgctgtaatttttgtagacaagttatgccat aagatgcttgtaatatattatggaaagaattttctgtaacaaatctcaggctcagcacctatggctctacatgt ttgtaatattaacattcgtattgtatcagtcaaaaaaacattcgtattggaagattctagttatacattaactg cagcegtgtccgattagcttcaagtcatagtagaaatttaagtgtaatattttaccttctgtaaaataaagttt aatgttttaccttttgtaaaaaaaaatgtaatattttagcatgcagtttagattaatttgtataacccaaaaat tgtcactcgatttaatttttgggttaaatcaagattttttttaatacgattgggectetttatttggcatgctgg ctaaattgggtetetttgt</INSDSeq sequences 34 </INSD&eg>

33 </SequenceData> 38 <Hedgquencelata seguanoelhNumbag="2vs 27 <INSDSeq> 28 <INSDSeq length»990</INSDSeq length» 38 ZINSDSegq molitypes>DNAC/INSDSeq moltyper> <IN3DSeq divisior>PAT</INSDIeqg division»

AL <INSDSeq feature-table> dz <IN3DFesture> 43 <INSDFeature keyvsource“/INSDFeature key> 44 “INSDFeature lecation>l..990</INSDFasature location» <INSDFeature quels» 4a <INSDOualifier> 477 <IN3DQualifier name>mol type</INSDQualifisr name> 48 <INSDQualifiler valuerother DNA</INGDGualifier value» 4% </INSDQualifier> <INSDQualifier id="gs">

SL CINSDQualifisr name>note</INSDQvali fier name> 57 <INSDQualifier value>CDS sequence of the gene

Gmygl2</IN3PQualifier value> 53 </INSDQualifier> 54 <INSDQualiflisr id="g3"> <IN3DQualifier namerorganism“/INSDQuali fier name>

LE <INSDQualifier valuersynthetic construct</INSDQualifier valued 57 </INSDOualiLfier»> 5a </INSDFeaturs guals> 53 </TNSDFeaturer a0 </INBDSeq feature-table> <INSDSex seduenceratgggttttagaattagcggttcctcctettcatcatgettttgtttgcaacgtac gettettcctecgttaacagcaatagccacttccactcegtggcttcaagaggaggagtaattgcataccgagtt gttcagtatcttatcgcaagtttgttgagtttgectttggatgaaaccacactccacactcacttgatccctteg cctttacaggaaaagtacaatttcatgaattccaaggatggtaaaggaactcttagtatgctatcatttgaagg tgccaaaattaggcttctacgaagtttgatcattgagacagaaacaatgcaggttttggattttactgtettte caaaagcagaatatgacatacccatattttgtgctaactttttcacctctgctaaaacaaacattgttgtgttg gaccttaaccccttgcatgatatcatcaatcagcatgagtacaaggagaagtactttaaaagcttaattcctect cggccttaaatatgctgagctttttccatggggagggaagctcacaagtgagtccataaattttttttcaccaa ttgtcatctggacaaagtttacctcaaacccagaaaaatatgatattetgtattccgcatttagggaatattac aaggtatggttgaaattgatatgcaaagcagataaagagacagatgaatctcagattttccacaacctcgaagc acaacatagatatctaacatggagagttgaaaaggatccaggacaaggtgttttgaagaagctgattggtgaca cacttgccaaggatatgctgagaagctttetctttaatggagttgatgaactaggaagcaaaacattcaatgat tattttccacgctactgctgtcaagagggaactctaaataaaaaaggcaatgttattgggaagtcctttgaaaa tcgcccatggaatgctagaggagaatttattgaatattttaggtga-/INSD5ed sequence» a2 </INSDSeg> a3 </SeguencaData> od <SequenceData sequenceIDNumber="3i> eo <INSDSeqr oe <INSDSeq length»>329</IN3DSeq length» 67 <“INSDSeqg moltype>AA</INSDSeg moltype> ad <INSDSeq divislion»PAT</INSDSeqg division» a3 <IN3D3eq feature-table> 0 <INSDEeature>

JL <IN3DFeature key>source</IN3DFeature key> ia <IN3DFeature locaticn>l..329</INSDFeaturs location» 73 <INSDFsature qualsg> 74 <INSDuuelifier>

TE <INSDoualifier name>mol type“ /INSDQualifier name>

TE <INSDQualifier value>protein</INSDQualifier value» 77 </INSDOualifier> in <INSDOuaiifier id="qg8*> ia <IN3DQualifier name>note</INSDQualifier name> 50 <INSDQualifiler valuerAmino acid sequence of the protein encoded by the CDS sequence</INSDQualifier valiue> 51 «</INSDOQualifier»>

SE <INSDQualifler ia="g5"> 83 <INSDQualifier name>organism</INSDQualifier name>

Sá <IN3DQualifier value>synthetic construct</INSDQualifier value» a5 </INSDOQuali fier

Se </INSDFsature guals> 57 </INSDFeature> 53 </INSDSeg feature-tabhle>

<INSDSeq sequence>MGFRISGSSSSSCFCLORTLLPPLTAIATSTRGFKRRSNCIPSCSVSYRKFVEFAL

DETTLHTHLIPSPLQEKYNFMNSKDGKGTLSMLSFEGAKIRLLRSLIIETETMQVLDFTVFPKAEYDIPIFCAN

FFTSAKTNIVVLDLNPLHDIINQHEYKEKYFKSLIPLGLKYAELFPWGGKLTSESINFFSPIVIWTKFTSNPEK

YDILYSAFREYYKVWLKLICKADKETDESQIFHNLEAQHRYLTWRVEKDPGOGVLKKLIGDTLAKDMLRSFLFN

GVDELGSKTFNDYFPRYCCQEGTLNKKGNVIGKSFENRPWNARGEFIEYFR</INSDSeq sequence

SG </INSDSeg>

Gl </Zequencebata>

Gi <SequernceData seguancaTiDNunbhao="4%> 33 <INSDSeqg> 34 <INSDSeq length>980</INSDSeq lengih> <INSDSeq moltype>DNA-/INSDSeg moltype>

Ga ZINSDSeq division>PAT</INSDSeq division»

GY <INSDSeq feabure-table> os <“INSDPearure> ad <INSDFeature key>source</INSDFeature keys

LOG <INSDFeature location>l..980</INSDFesature locations

Ol <INSDFealurse guals> 102 <INSDOQualifier» 142 <IN3DQualifier namedmol type</INSDQualifisr name> 104 <INSDQualifier value>other DNA</IN3DGualifier value> 10% </INSDQuali fier»

Lod <INSDQuaiifler id="g8*>

Ee <INSDOQualifier namernote</INSDQualifiesr name> ies <INSDQualiflisr valverGmygl2 sequence of the ygl2 mutant</INSDQualifier valuer 109 </INSDOualifier> iin <INSDOualiifier id="qg"*> iid <IN3DQualifier name>organism</INSDQualifisr name>

Li <INSDQualifier valuersynthetic construct</INSDQualifier valued

LLS «</INSDOQualifier»> 114 </INSDFeaturs auals> 115 </INSDFsature> 118 “/INSDSeg feature-table> <IN3D3eq sequence>atgggttttagaattagcggttectcctcettcatcatgettttgtttgcaacgtac gcttettecteegttaacagecaatagecacttecactegtggettcaagaggaggagtaattgecatacegagtt gttcagtatcttatcgcaagtttgttgagtttgectttggatgaaaccacactccacactcacttgatccctteg cctttacaggaaaagtacaatttcatgaattccaaggatggtaaaggaactcttaatttgaaggtgccaaaatt aggcttctacgaagtttgatcattgagacagaaacaatgcaggttttggattttactgtctttccaaaagcaga atatgacatacccatattttgtgctaactttttcacctctgctaaaacaaacattgttgtgttggaccttaacc ccttgcatgatatcatcaatcagcatgagtacaaggagaagtactttaaaagcttaattcctcteggeccttaaa tatgctgagetttttccatggggagggaagctcacaagtgagtccataaattttttttcaccaattgtcatctg gacaaagtttacctcaaacccagaaaaatatgatattctgtattcecgcatttagggaatattacaaggtatggt tgaaattgatatgcaaagcagataaagagacagatgaatctcagattttccacaacctcgaagcacaacataga tatctaacatggagagttgaaaaggatccaggacaaggtgttttgaagaagctgattggtgacacacttgccaa ggatatgctgagaagctttctctttaatggagttgatgaactaggaagcaaaacattcaatgattattttccac gctactgctgtcaagagggaactctaaataaaaaaggcaatgttattgggaagtcctttgaaaatcgcccatgg aatgctagaggagaatttattgaatattttaggtga/INSDS=4 zeguence> iis </INSDSeg> iis </SeguenceData> 120 <SeguenceData semuenceIDNumber="Bn> 124 <INSDSeq»

Lal <INSDSeq lengih>988</INSD53eq lengths

Las <INSDSeq moltype>AA</INSDSeq moltype> 124 <INSDSeq division>PAT</INSDSeq division» 125 <INSDSeq feature-iable> ize <INSDFeaturer» 1a <IN3DFeature key>source</IiN3DFeature key»

Le <IN3DFeature lowation>l..98</INSDFeaturs Locations» 12% <INSDFeature guals> 130 <INSDOualifier>

TEL <INSDOQualifier name>mol type</INSDQualifier name>

LS <“INSDoualifier valuesprotein</INSDQualifier value» 133 </INSDOualifier> 134 <INSDOualifier id="qL0%> ijb <IN3DQualifier namesnote</1NSDQualifier name>

Lie <INSDQualifier value>Amino acid sequence of the protein encoded by Gmygl2 from the ygl2 mitant</INSEQualifier value»

RT </INSDOualifier>

L3H <INSDQualifler ia=’gS"> 13% <INSDQualifier name>organism</INSDQualifier name> 140 <IN3DQualifier value>synthetic construct</INSDQualifier value» 14d </INSDOQuali fier 142 </INSDFesature duals» 143 </INSDFeature> 144 </INSDSeg feature-table> <INSDSed sequsnce>MGFRISGSSSSSCFCLQORTLLPPLTAIATSTRGFKRRSNCIPSCSVSYRKFVEFAL

DETTLHTHLIPSPLQEKYNFMNSKDGKGTLNLKVPKLGFYEV</INSDSeqg sequence’ 148 </INSDSec> u 1a </SeguencaData> 145 <SequenceData sequenceIDNumber="6n> ide <INSDSeqr a0 <INSDSeq length»>22</INSDSeqg length»

Lel <INSDSeq moltype>DNA</INSDSeg moltype> isd <INSDSeq divislion»PAT</INSDSeqg division» is <INSDSeg Iearturertabie» 154 <INSDPsaturer 155 <IN3DFeature key>source</IN3DFeature key> ihe <IN3DFeature location>»l..22</INSDFeaturs locations

Ls? <INSDFsature qualsg>

LAE <INSDuuelifier>

LDS <INSDoualifier name>mol type“ /INSDQualifier name> 164 <INSDQualifisr value>other DNA</INSDQualifier valuer 161 </INSDOualifier> 182 <INSDOualifier id="qgiar> 123 <IN3DQualifier name>note</INSDQualifier name> ied <INSDQualifier valuerForward primer for

ID105</INsDgualifier value»

TED </INSDOualifier>

LEE <INSDQualifler ia="gil> 167 <INSDQualifier name>organism</INSDQualifier name> 158 <IN3DQualifier value>synthetic construct /INSDQualifier values

TAQ </THNSDQualifiszr> u 170 </INSDFesature duals»

LIL </INSDFeature>

Tis </INSDSeg feature-table>

HR <INSDSeq sequance>aagagttttactatgactatgt</INSDSeq sequences 174 </INSDS so» 175 </Seguencedata> 1a <SequenceData sequence lDNumben="THs iT <INSDSeq> 178 <INSDSeq lengibh»22</INSDSeq length

Lo <INSDSeq moltype>DNA</INSDSeg moltype»

LEO <INSDSeq division>PAT</INSDSeg division» iel <INSDSeq feature-table>

LS? <INSDFeature> 182 <INSDFeature key>sourcec/INSDFeature key» 184 <INSDFearure location>l..22</INSDFeature location» 185 <INSDFeature guals> isa <INSDOualifien>

RY <INSDQualifier name>mol type</INSDQualiifier name>

LEE <INSDQualifier valuerother DNA</INSDQualifier value» 155 </INSDOualiLfier»> 194 <INSDQualiifler id="glá"> 191 <INSDQualifier name>note</INSDQualifier name> 152 <IN3DQualifier value>Reverse primer for

ID105</INSDCuaiifier value» 193 </INSDQuali fier» u 184 <INSDQuaiifier id="gijx>

Lan <INSDOQualifier namerorganism</INSDQualifier name>

Lan <INSDQualiflsr value>synthetic construct /INSDQualifier value> 197 </INSDOualifier> 138 </IN3DFeature guala> iss </INSDFeaturer u 204 </INSDSegy featurs-table> 201 <INSDSeq sequence>ataaatacagacaaagtcaaat</INSDSey sequences

ZOL </INSDSeg>

“03 </SequenceData>

Zid “<SequenceData segusnceliNumec=NS"> 205 <INSDSeqg> 208 <IN3DSeq length»21</INSDSeq length» 207 ZINSDSegq molitypes>DNAC/INSDSeq moltyper> 208 <IN3DSeq divisior>PAT</INSDIeqg division» 20% <INSDSeq feature-table> 210 <INS3DFesature> all <INSDFeaturs keyrsource</INSDFeaturs key> 217 <INSDFeature location>l..21</IN3DFeature location» zi <INSDhFeature quels» 214 <INSDQualifier> 215 <IN3DQualifier name>mol type</INSDQualifisr name> 21a <INSDQualifiler valuerother DNA</INGDGualifier value» 217 </INSDQualifier> zis <INSDQualifier id="gióx>

PARE CINSDQualifisr name>note</INSDQvali fier name>

ZL <INSDQualifier valus>Forward primer for PCR amplication of RNA from GLl11</INSDQualifier value» 22 </INSDQualifier> u 222 <INSDOualifier Ld=Yglin> 223 <INSDQualifier namerorganism“/INSDQuali fier name> 224 <INSDQualifier valuersynthetic construct</INSDQualifier valued a </INSDOualifiers

LEG </INSDFeaturs guals>

Ze </TNSDFeaturer 228 </INBDSeq feature-table> 22% <IN3DSeqg sequenzs>atgggttttagaattageggt-/INSDSeq sequences 2340 </INSDSeg> 231 </Zequencebatar

EAR <Sequencebata seguenasibNumbar=snats

HSE <IN3D3eq> 234 <INSDSeq length>26</IN3DSeq length» 235 <INSDSeq moltype>DNA</INSDSeq moltype> 238 <IN3DSeq division»PAT</INSD3eq division» 237 <INSDSeq fearure-tabier 238 <INSDFeabture> 230 <INSDFeature key>source“/INSDFeature key>

ZAG <INSDFeature location>l..26</INSDFeature location> “4% <INSDFeaturs quals> u da? <INSDQualifier»> dz <INSDQualifier name>mol type</iNSDQualifier name> 244d <INSDuuelifier value>other DNA</INSDOualifier valued 245 </THSDQualifise> u 2448 <INSDQualifier id="giënx> 247 <INSDQualifier namernote</INSDQualifier named

ZA <INSDQualifier valuerReverse primer for PCR amplication of RNA from GL11</INSDPQualifisr value»

PAG </INSDQualifiers> 250 <INSDOualifier id="qL7n> 251 <IN3DQualifier namerorganism</INSDQualifiesr name> 252 <INSDQualifier valuersynthetic construct</INSDoualifier value» 25% </INSDQualifiers> u 254 </INSDFeature quals> wss </IN3DFeature> u 258 “/INSDSeqg fesature-table> 257 <INSDSeg sequencercctaaaatattcaataaattetecte</INSDSey sequences 258 </INSDSeq> 7 258 </SeguenceDatas 288 <SequenceData zagquencalDNumber="310"> zel <INSDSeqr

EA <INSDSeq length>63</INSD5eq length» “ES <“INSDSeqg moltype>AA</INSDSeg moltype>

Z64 <INSDSeq divislion»PAT</INSDSeqg division» 25h <INSDSeg feature~tablex 260 <INSDFeature> 267 <IN3DFeature key>source</IN3DFeature key> 288 <IN3DFeature location>»l..63</INSDFeaturs locations 28% <INSDFsature qualsg> <INSDuuelifier>

EL <INSDoualifier name>mol type“ /INSDQualifier name>

Eid <INSDQualifier value>protein</INSDQualifier value» 272 </INSDOualifier> 27d <INSDOualifier id="gq20%> 275 <IN3DQualifier name>note</INSDQualifier name> 278 <INSDQualifiler valuerAmino acid sequence of the protein encoded by the gene Gmygl2 from

YGL2-SP1</INSDQuali fier values

Aa </INSDOualifiers

AES <INSDQualiifler id="gLS8*>

Et <INSDQualifier name>organism</iNSDQualifier name> 280 <IN3DQualifier value>synthetic construct</INSDQualifier valued» 281 </INSDQuali fier» 282 </INSDFearure quals>

ZES «/INSDFeaturer

WEE! </INSDSeg feature-table> <INSDSeq sequence>MGFRISGSSSSSCFCLQRTLLPPLTAIATSIVASRGGVIAYRVVOYLASLLSLLWM

KPHSTLT</INSDSeq sequence» 288 </TNSDSeg> 287 </SeguenceDala> 288 </8TZeSequencelisting>

Claims (7)

Conclusions l. Gene Gmygl2 related to soybean plant height and leaf color, having a nucleotide sequence comprising a) or b): a) a nucleotide sequence shown in SEQ ID NO: 1; and b) a nucleotide sequence formed by performing substitution, deletion or addition of bases on the nucleotide sequence shown in SEQ ID NO: 1 and having a similar function to the nucleotide sequence shown in SEQ ID NO: 1. A protein encoded by the gene Grmyg!2 according to claim 1, having an amino acid sequence shown in SEQ ID NO: 3. 3. Insertion-deletion (InDel) marker linked to a gene Gmiygl2 related to soybean plant height and leaf color, where primers of the InDel marker are an upstream primer InDel-105-F and a downstream primer InDel-105-R include; wherein the upstream primer InDel-105-F has a nucleotide sequence comprising any of 1)-3): 1) a nucleotide sequence shown in SEQ ID NO: 6; 2) a nucleotide sequence formed by performing substitution, deletion or addition of bases on the nucleotide sequence shown in SEQ ID NO: 6 and having a similar function to the nucleotide sequence shown in SEQ ID NO: 6; and 3) a nucleotide sequence that is a derivative of the nucleotide sequence shown in SEQ ID NO: 6 and has a similar function to the nucleotide sequence shown in SEQ ID NO: 6; and wherein the downstream primer InDel-105-R has a nucleotide sequence comprising any of 4)-6): 4) a nucleotide sequence shown in SEQ ID NO: 7; 5) a nucleotide sequence formed by performing substitution, deletion or addition of bases on the nucleotide sequence shown in SEQ ID NO: 7 and having a similar function to the nucleotide sequence shown in SEQ ID NO: 7; and 6) a nucleotide sequence that is a derivative of the nucleotide sequence shown in SEQ ID NO: 7 and has a similar function to the nucleotide sequence shown in SEQ ID NO: 7. The InDel marker of claim 3, wherein the InDel marker is located on chromosome 2 of a soybean genome. The InDel marker of claim 3, wherein genes identified by the InDel marker include any of A1-A5: A1, the nucleotide sequence shown in SEQ ID NO: 1; A2, the nucleotide sequence formed by performing substitution, deletion or addition of bases on the nucleotide sequence shown in SEQ ID NO: 1 and having a similar function to the nucleotide sequence shown in SEQ ID NO: 1; A3, a nucleotide sequence comprising the nucleotide sequence shown in SEQ ID NO: 1; and A4, a nucleotide sequence comprising part of the nucleotide sequence shown in SEQ ID NO:1. Use of the gene Gmyyg!2 related to soybean plant height and leaf color according to claim 1 or the protein according to claim 2 or the InDel marker according to any of claims 3-5 in any of B1-B5: B1, identifying a leaf color and a plant height of a soybean; B2, preparing a product capable of identifying the leaf color and plant height of the soybean; B3, to assist in identifying soybean leaf color and plant height; B4, preparing a product that is able to assist in identifying the leaf color and plant height of the soybean; and BS, soybean cultivation. 7. Method for screening a soybean with a yellow-green leaf color and a desired plant height, wherein the method comprises the following steps: carrying out PCR amplification on a genomic DNA of a soybean to be screened using primers from the InDel marker according to any of claims 3-5, and selecting a material with a PCR amplification product of 149 bp, to obtain the soybean with a yellow-green leaf color and a desired plant height.