
Chinese Journal OF Rice Science ›› 2025, Vol. 39 ›› Issue (5): 624-634.DOI: 10.16819/j.1001-7216.2025.240106
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SHAO Ye1,2,#, HU Yuanyi1,2,#, PENG Yan1,2,#, MAO Bigang1,2, LIU Huimin1,2, TANG Chanjuan1, LEI Bin1,2, TANG Li1,2, YU Lixia3, LI Wenjian3, LUO Wuzhong3, LUO Zhibin1,2, YUAN Yuantao1,2, LI Yaokui1,2, ZHANG Dan1,2, ZHOU Libin3, BAI Lianyang2,4,*(
), TANG Wenbang1,2,*(
), ZHAO Bingran1,2,*(
)
Received:2024-01-09
Revised:2024-04-17
Online:2025-09-10
Published:2025-09-10
About author:#These authors contributed equally to this work
韶也1,2,#, 胡远艺1,2,#, 彭彦1,2,#, 毛毕刚1,2, 刘慧敏1,2, 唐婵娟1, 雷斌1,2, 唐丽1,2, 余丽霞3, 李文建3, 罗武中3, 罗治斌1,2, 袁远涛1,2, 李曜魁1,2, 张丹1,2, 周利斌3, 柏连阳2,4,*(
), 唐文帮1,2,*(
), 赵炳然1,2,*(
)
作者简介:#共同第一作者
基金资助:SHAO Ye, HU Yuanyi, PENG Yan, MAO Bigang, LIU Huimin, TANG Chanjuan, LEI Bin, TANG Li, YU Lixia, LI Wenjian, LUO Wuzhong, LUO Zhibin, YUAN Yuantao, LI Yaokui, ZHANG Dan, ZHOU Libin, BAI Lianyang, TANG Wenbang, ZHAO Bingran. Directed Improvement of Hybrid Rice Zhuoliangyou 1126 by Heavy Ion Beam Mutagenesis Based on M1TDS Targeted Screening Technology[J]. Chinese Journal OF Rice Science, 2025, 39(5): 624-634.
韶也, 胡远艺, 彭彦, 毛毕刚, 刘慧敏, 唐婵娟, 雷斌, 唐丽, 余丽霞, 李文建, 罗武中, 罗治斌, 袁远涛, 李曜魁, 张丹, 周利斌, 柏连阳, 唐文帮, 赵炳然. 基于M1TDS靶向筛选技术的重离子束诱变定向改良杂交水稻卓两优1126性状的研究[J]. 中国水稻科学, 2025, 39(5): 624-634.
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URL: http://www.ricesci.cn/EN/10.16819/j.1001-7216.2025.240106
| 名称 Name | 序列 Sequence |
|---|---|
| OsNRAMP5-246-8-F | GAAGGTGACCAAGTTCATGCTTGTGGTTGGCTCCGGCTTAA |
| OsNRAMP5-246-8-H | GAAGGTCGGAGTCAACGGATTTGTGGTTGGCTCCGGCTTGC |
| OsNRAMP5-246-8-C | CACAAAATGAAACAGTGGAAACCGATC |
| OsNRAMP5-234-6-F | GAAGGTGACCAAGTTCATGCTCACAAAATGAAACAGTGGAAACCGAT |
| OsNRAMP5-234-6-H | GAAGGTCGGAGTCAACGGATTCACAAAATGAAACAGTGGAAACCGAC |
| OsNRAMP5-234-6-C | CGTACCTCATATCTGTGGTTGGCTC |
| OsNRAMP5-204-6-F | GAAGGTGACCAAGTTCATGCTCACAAAATGAAACAGTGGAAACCGA |
| OsNRAMP5-204-6-H | GAAGGTCGGAGTCAACGGATTCACAAAATGAAACAGTGGAAACCGC |
| OsNRAMP5-204-6-C | CGTACCTCATATCTGTGGTTGGCTC |
| OsNRAMP5-37-7-F | GAAGGTGACCAAGTTCATGCTTGAAGAAGCTAAGAGAGGAAGCAACAA |
| OsNRAMP5-37-7-H | GAAGGTCGGAGTCAACGGATTTGAAGAAGCTAAGAGAGGAAGCAACAG |
| OsNRAMP5-37-7-C | ACTAGCTCTCCAGCTGATGCTC |
| OsBADH2-168-1-F | GAAGGTGACCAAGTTCATGCTGTGTAGTTGGGTTGATCACACCTT |
| OsBADH2-168-1-H | GAAGGTCGGAGTCAACGGATTGTGTAGTTGGGTTGATCACACCTC |
| OsBADH2-168-1-C | TGGGTCATAAATAAATATAAGCGCAGG |
| OsBADH2-224-8-F | GAAGGTGACCAAGTTCATGCTTTCGTCTTTTCTTGACAGCCTGTT |
| OsBADH2-224-8-H | GAAGGTCGGAGTCAACGGATTTTCGTCTTTTCTTGACAGCCTGTC |
| OsBADH2-224-8-C | AGGACTTTTTCCACCAAGTTCCAG |
| OsLOX3-760-7-F | GAAGGTGACCAAGTTCATGCTTATCTTCACCTATGCCACAAGGC |
| OsLOX3-760-7-H | GAAGGTCGGAGTCAACGGATTTATCTTCACCTATGCCACAAGGA |
| OsLOX3-760-7-C | CAGGGTATCATCATCACGTAAGAAC |
| OsLOX3-115-5-F | GAAGGTGACCAAGTTCATGCTGAGAGCAAAGTGTTGAACATGAAC |
| OsLOX3-115-5-H | GAAGGTCGGAGTCAACGGATTGAGAGCAAAGTGTTGAACATGAAG |
| OsLOX3-115-5-C | GGACCGACCCGGTTCTTGAG |
| OsPAO5-549-5-F | GAAGGTGACCAAGTTCATGCTTGCTTGACAGGAATCCACACCTA |
| OsPAO5-549-5-H | GAAGGTCGGAGTCAACGGATTTGCTTGACAGGAATCCACACCTG |
| OsPAO5-549-5-C | GCACCACCATTCCAGATATAGGTG |
| OsPAO5-1028-4-F | GAAGGTGACCAAGTTCATGCTAACTTTCCTCCAGGGTTACCAAAAT |
| OsPAO5-1028-4-H | GAAGGTCGGAGTCAACGGATTAACTTTCCTCCAGGGTTACCAAATC |
| OsPAO5-1028-4-C | GCTTGTCCCATCTTCAACACATAC |
| OsPAO5-84-2-F | GAAGGTGACCAAGTTCATGCTCTCAGCTCAAGAAAATGCTACCAGG |
| OsPAO5-84-2-H | GAAGGTCGGAGTCAACGGATTCTCAGCTCAAGAAAATGCTACCACT |
| OsPAO5-84-2-C | GACAGTACTCAACTGTACATGGTCTG |
| OsSSIIIa-598-7-F | GAAGGTGACCAAGTTCATGCTCCATTTGGTTCAAGGCCTAGAACTC |
| OsSSIIIa-598-7-H | GAAGGTCGGAGTCAACGGATTCCATTTGGTTCAAGGCCTAGAACTG |
| OsSSIIIa-598-7-C | CACTGTTTTCGACGTAGACCATG |
| OsBEIIb-931-5-F | GAAGGTGACCAAGTTCATGCTGTGTGATTGTTATTAAATCTCACCAGGA |
| OsBEIIb-931-5-H | GAAGGTCGGAGTCAACGGATTGTGTGATTGTTATTAAATCTCACCAGGC |
| OsBEIIb-931-5-C | GGCTATCTTAACTTTATGGGAAATGAGTTC |
Table 1. Primer sequence for KASP genotyping
| 名称 Name | 序列 Sequence |
|---|---|
| OsNRAMP5-246-8-F | GAAGGTGACCAAGTTCATGCTTGTGGTTGGCTCCGGCTTAA |
| OsNRAMP5-246-8-H | GAAGGTCGGAGTCAACGGATTTGTGGTTGGCTCCGGCTTGC |
| OsNRAMP5-246-8-C | CACAAAATGAAACAGTGGAAACCGATC |
| OsNRAMP5-234-6-F | GAAGGTGACCAAGTTCATGCTCACAAAATGAAACAGTGGAAACCGAT |
| OsNRAMP5-234-6-H | GAAGGTCGGAGTCAACGGATTCACAAAATGAAACAGTGGAAACCGAC |
| OsNRAMP5-234-6-C | CGTACCTCATATCTGTGGTTGGCTC |
| OsNRAMP5-204-6-F | GAAGGTGACCAAGTTCATGCTCACAAAATGAAACAGTGGAAACCGA |
| OsNRAMP5-204-6-H | GAAGGTCGGAGTCAACGGATTCACAAAATGAAACAGTGGAAACCGC |
| OsNRAMP5-204-6-C | CGTACCTCATATCTGTGGTTGGCTC |
| OsNRAMP5-37-7-F | GAAGGTGACCAAGTTCATGCTTGAAGAAGCTAAGAGAGGAAGCAACAA |
| OsNRAMP5-37-7-H | GAAGGTCGGAGTCAACGGATTTGAAGAAGCTAAGAGAGGAAGCAACAG |
| OsNRAMP5-37-7-C | ACTAGCTCTCCAGCTGATGCTC |
| OsBADH2-168-1-F | GAAGGTGACCAAGTTCATGCTGTGTAGTTGGGTTGATCACACCTT |
| OsBADH2-168-1-H | GAAGGTCGGAGTCAACGGATTGTGTAGTTGGGTTGATCACACCTC |
| OsBADH2-168-1-C | TGGGTCATAAATAAATATAAGCGCAGG |
| OsBADH2-224-8-F | GAAGGTGACCAAGTTCATGCTTTCGTCTTTTCTTGACAGCCTGTT |
| OsBADH2-224-8-H | GAAGGTCGGAGTCAACGGATTTTCGTCTTTTCTTGACAGCCTGTC |
| OsBADH2-224-8-C | AGGACTTTTTCCACCAAGTTCCAG |
| OsLOX3-760-7-F | GAAGGTGACCAAGTTCATGCTTATCTTCACCTATGCCACAAGGC |
| OsLOX3-760-7-H | GAAGGTCGGAGTCAACGGATTTATCTTCACCTATGCCACAAGGA |
| OsLOX3-760-7-C | CAGGGTATCATCATCACGTAAGAAC |
| OsLOX3-115-5-F | GAAGGTGACCAAGTTCATGCTGAGAGCAAAGTGTTGAACATGAAC |
| OsLOX3-115-5-H | GAAGGTCGGAGTCAACGGATTGAGAGCAAAGTGTTGAACATGAAG |
| OsLOX3-115-5-C | GGACCGACCCGGTTCTTGAG |
| OsPAO5-549-5-F | GAAGGTGACCAAGTTCATGCTTGCTTGACAGGAATCCACACCTA |
| OsPAO5-549-5-H | GAAGGTCGGAGTCAACGGATTTGCTTGACAGGAATCCACACCTG |
| OsPAO5-549-5-C | GCACCACCATTCCAGATATAGGTG |
| OsPAO5-1028-4-F | GAAGGTGACCAAGTTCATGCTAACTTTCCTCCAGGGTTACCAAAAT |
| OsPAO5-1028-4-H | GAAGGTCGGAGTCAACGGATTAACTTTCCTCCAGGGTTACCAAATC |
| OsPAO5-1028-4-C | GCTTGTCCCATCTTCAACACATAC |
| OsPAO5-84-2-F | GAAGGTGACCAAGTTCATGCTCTCAGCTCAAGAAAATGCTACCAGG |
| OsPAO5-84-2-H | GAAGGTCGGAGTCAACGGATTCTCAGCTCAAGAAAATGCTACCACT |
| OsPAO5-84-2-C | GACAGTACTCAACTGTACATGGTCTG |
| OsSSIIIa-598-7-F | GAAGGTGACCAAGTTCATGCTCCATTTGGTTCAAGGCCTAGAACTC |
| OsSSIIIa-598-7-H | GAAGGTCGGAGTCAACGGATTCCATTTGGTTCAAGGCCTAGAACTG |
| OsSSIIIa-598-7-C | CACTGTTTTCGACGTAGACCATG |
| OsBEIIb-931-5-F | GAAGGTGACCAAGTTCATGCTGTGTGATTGTTATTAAATCTCACCAGGA |
| OsBEIIb-931-5-H | GAAGGTCGGAGTCAACGGATTGTGTGATTGTTATTAAATCTCACCAGGC |
| OsBEIIb-931-5-C | GGCTATCTTAACTTTATGGGAAATGAGTTC |
| 名称 Name | 序列 Sequence |
|---|---|
| OsNRAMP5-3E-540F | GTTGGTCCTGGATTCATGGTGTC |
| OsNRAMP5-3E-540R | TCCAATCAGAATCACCCAGAGCAG |
| OsNRAMP5-1E-265F | TCTTCGTCTACTTCCAGCTAGCC |
| OsNRAMP5-1E-265R | ACAGTGAACAACTGTCCATGGTG |
| OsBADH-4-471F | TTGATGAAGCAGCATGGGACATG |
| OsBADH-4-471R | GCTGCTAGGTACAATTTGTGAGAC |
| OsBADH-8-357F | CTTCAGCTGCTCCTATGGTTAAGG |
| OsBADH-8-357R | TCTAGCATCCAGCTCAGTTAAGTGC |
| OsLOX3-8-535F | GAGAAGAACCTCGAAGGCCTCAG |
| OsLOX3-8-535R | CCATCACAGCATGTGTGTTGAGC |
| OsPAO-6/7-563F | GCTCTCTTTGATAAGGATGGTCGTC |
| OsPAO-6/7-563R | ATGGCCACCAGTAAGAACATGTTC |
| OsBEIIb-17-359F | GGCATGAGAACCTCACATACTG |
| OsBEIIb-17-359R | GGAAGTACTTGTGGAGCTCTTGG |
Table 2. Primers for Sanger sequencing
| 名称 Name | 序列 Sequence |
|---|---|
| OsNRAMP5-3E-540F | GTTGGTCCTGGATTCATGGTGTC |
| OsNRAMP5-3E-540R | TCCAATCAGAATCACCCAGAGCAG |
| OsNRAMP5-1E-265F | TCTTCGTCTACTTCCAGCTAGCC |
| OsNRAMP5-1E-265R | ACAGTGAACAACTGTCCATGGTG |
| OsBADH-4-471F | TTGATGAAGCAGCATGGGACATG |
| OsBADH-4-471R | GCTGCTAGGTACAATTTGTGAGAC |
| OsBADH-8-357F | CTTCAGCTGCTCCTATGGTTAAGG |
| OsBADH-8-357R | TCTAGCATCCAGCTCAGTTAAGTGC |
| OsLOX3-8-535F | GAGAAGAACCTCGAAGGCCTCAG |
| OsLOX3-8-535R | CCATCACAGCATGTGTGTTGAGC |
| OsPAO-6/7-563F | GCTCTCTTTGATAAGGATGGTCGTC |
| OsPAO-6/7-563R | ATGGCCACCAGTAAGAACATGTTC |
| OsBEIIb-17-359F | GGCATGAGAACCTCACATACTG |
| OsBEIIb-17-359R | GGAAGTACTTGTGGAGCTCTTGG |
| 目标基因 Target gene | 基因编号 Gene ID | 目标表型 Target phenotype | 外显子数 No. of exons | 目标区域覆盖度 Coverage of target area(%) | 平均测序深度 Average sequencing depth |
|---|---|---|---|---|---|
| OsNRAMP5 | Os07g0257200 | 镉低积累 Low cadmium accumulation | 13 | 100 | 53398× |
| OsBADH2 | Os08g0424500 | 香味Aroma | 15 | 100 | 53086× |
| OsLOX3 | Os03g0699700 | 耐储藏Storage-tolerance | 9 | 100 | 50202× |
| OsPAO5 | Os04g0671300 | 耐淹 Submergence-tolerance | 10 | 100 | 54263× |
| OsSSIIIa | Os08g0191433 | 低升糖指数 Low glycemic index (GI) | 14 | 100 | 54510× |
| OsBEIIb | Os02g0528200 | 低升糖指数 Low glycemic index (GI) | 22 | 100 | 52192× |
Table 3. Target gene and targeted sequencing analysis
| 目标基因 Target gene | 基因编号 Gene ID | 目标表型 Target phenotype | 外显子数 No. of exons | 目标区域覆盖度 Coverage of target area(%) | 平均测序深度 Average sequencing depth |
|---|---|---|---|---|---|
| OsNRAMP5 | Os07g0257200 | 镉低积累 Low cadmium accumulation | 13 | 100 | 53398× |
| OsBADH2 | Os08g0424500 | 香味Aroma | 15 | 100 | 53086× |
| OsLOX3 | Os03g0699700 | 耐储藏Storage-tolerance | 9 | 100 | 50202× |
| OsPAO5 | Os04g0671300 | 耐淹 Submergence-tolerance | 10 | 100 | 54263× |
| OsSSIIIa | Os08g0191433 | 低升糖指数 Low glycemic index (GI) | 14 | 100 | 54510× |
| OsBEIIb | Os02g0528200 | 低升糖指数 Low glycemic index (GI) | 22 | 100 | 52192× |
| 目标基因 Target gene | 亲本 Parent | 混池编号 No. of the pool | 突变位置 Mutation site | 野生/突变基因型 Wild/mutant genotype |
|---|---|---|---|---|
| OsNRAMP5 | 卓234S Zhuo 234S | 217-288E | 第3外显子Exon 3 | C/CTTAA |
| 卓234S Zhuo 234S | 217-288W | 第3外显子Exon 3 | C/CTTAA | |
| 卓234S Zhuo 234S | 217-288E | 第3外显子Exon 3 | GA/G | |
| 湘农恢1126 Xiangnonghui 1126 | 145-216S | 第3外显子Exon 3 | GCAGAT/G | |
| 湘农恢1126 Xiangnonghui 1126 | 1-72N | 第1外显子Exon 1 | CTCAATCTCCAT/C | |
| OsBADH2 | 卓234S Zhuo 234S | 145-216E | 第4外显子和第4内含子Exon 4/Intron 4 | CCTTGGTATTTCACATTTTTCT/C |
| 湘农恢1126 Xiangnonghui 1126 | 217-238S | 第8外显子Exon 8 | GTT/G | |
| OsLOX3 | 卓234S Zhuo 234S | 718-789W | 第7外显子Exon 7 | G/GC |
| 湘农恢1126 Xiangnonghui 1126 | 73-144S | 第9外显子Exon 9 | GAAC/G | |
| 湘农恢1126 Xiangnonghui 1126 | 73-144E | 第9外显子Exon 9 | GAAC/G | |
| OsPAO5 | 卓234S Zhuo 234S | 504-575W | 第6外显子和第6内含子Exon 6/Intron 6 | CACTTACTTT/C |
| 卓234S Zhuo 234S | 1004-1040E | 第9外显子Exon 9 | ATGAT/A | |
| 卓234S Zhuo 234S | 991-1040W | 第9外显子Exon 9 | ATGAT/A | |
| 湘农恢1126 Xiangnonghui 1126 | 73-144E | 第9外显子Exon 9 | GTAGCTCC/G | |
| OsSSIIIa | 卓234S Zhuo 234S | 576-647E | 第13外显子Exon 13 | CTCG/C |
| OsBEIIb | 卓234S Zhuo 234S | 863-932W | 第17外显子Exon 17 | GATGT/G |
Table 4. Mutations were captured by mixed-pool targeted gene sequencing
| 目标基因 Target gene | 亲本 Parent | 混池编号 No. of the pool | 突变位置 Mutation site | 野生/突变基因型 Wild/mutant genotype |
|---|---|---|---|---|
| OsNRAMP5 | 卓234S Zhuo 234S | 217-288E | 第3外显子Exon 3 | C/CTTAA |
| 卓234S Zhuo 234S | 217-288W | 第3外显子Exon 3 | C/CTTAA | |
| 卓234S Zhuo 234S | 217-288E | 第3外显子Exon 3 | GA/G | |
| 湘农恢1126 Xiangnonghui 1126 | 145-216S | 第3外显子Exon 3 | GCAGAT/G | |
| 湘农恢1126 Xiangnonghui 1126 | 1-72N | 第1外显子Exon 1 | CTCAATCTCCAT/C | |
| OsBADH2 | 卓234S Zhuo 234S | 145-216E | 第4外显子和第4内含子Exon 4/Intron 4 | CCTTGGTATTTCACATTTTTCT/C |
| 湘农恢1126 Xiangnonghui 1126 | 217-238S | 第8外显子Exon 8 | GTT/G | |
| OsLOX3 | 卓234S Zhuo 234S | 718-789W | 第7外显子Exon 7 | G/GC |
| 湘农恢1126 Xiangnonghui 1126 | 73-144S | 第9外显子Exon 9 | GAAC/G | |
| 湘农恢1126 Xiangnonghui 1126 | 73-144E | 第9外显子Exon 9 | GAAC/G | |
| OsPAO5 | 卓234S Zhuo 234S | 504-575W | 第6外显子和第6内含子Exon 6/Intron 6 | CACTTACTTT/C |
| 卓234S Zhuo 234S | 1004-1040E | 第9外显子Exon 9 | ATGAT/A | |
| 卓234S Zhuo 234S | 991-1040W | 第9外显子Exon 9 | ATGAT/A | |
| 湘农恢1126 Xiangnonghui 1126 | 73-144E | 第9外显子Exon 9 | GTAGCTCC/G | |
| OsSSIIIa | 卓234S Zhuo 234S | 576-647E | 第13外显子Exon 13 | CTCG/C |
| OsBEIIb | 卓234S Zhuo 234S | 863-932W | 第17外显子Exon 17 | GATGT/G |
Fig. 1. KASP genotyping in the mixed pool of target gene mutations A-D represent the genotyping of different mutation sites of OsNRAMP5 gene; E and F represent the genotyping of different mutation sites of OsBADH2 gene, respectively; G and H represent genotyping of different mutation sites of OsLOX3 gene, respectively; I-K represent genotyping of different mutation sites of OsPAO5 gene; L, M represent the genotyping of mutation sites in OsSSIIIa gene and OsBEIIb gene, respectively; In panel A, 246-8 represents the 8th plant in 246 rows of M1 population (the same below); Blue/green dots represent wild genotypes, red dots represent mutant genotypes, and gray dots represent negative controls; FAM and HEX represent fluorescently modified labels of two different colors, respectively.
| 基因 Gene | 品种 Variety | 突变株定位(行-株) Location of mutants (row-plant) | 突变分蘖占比 Proportion of tillers containing mutations |
|---|---|---|---|
| OsNRAMP5 | 卓234S Zhuo 234S | 246-8 | 18/18 |
| OsNRAMP5 | 卓234S Zhuo 234S | 234-6 | 4/18 |
| OsNRAMP5 | 湘农恢1126 Xiangnonghui 1126 | 204-6 | 3/6 |
| OsNRAMP5 | 湘农恢1126 Xiangnonghui 1126 | 37-7 | 1/8 |
| OsBADH2 | 卓234S Zhuo 234S | 168-1 | 3/18 |
| OsBADH2 | 湘农恢1126 Xiangnonghui 1126 | 224-8 | 1/6 |
| OsLOX3 | 卓234S Zhuo 234S | 760-7 | 1/18 |
| OsLOX3 | 湘农恢1126 Xiangnonghui 1126 | 115-5 | 2/6 |
| OsPAO5 | 卓234S Zhuo 234S | 549-5 | 1/18 |
| OsPAO5 | 卓234S Zhuo 234S | 1028-4 | 5/18 |
| OsPAO5 | 湘农恢1126 Xiangnonghui 1126 | 84-2 | 1/6 |
| OsSSIIIa | 卓234S Zhuo 234S | 598-7 | 1/18 |
| OsBEIIb | 卓234S Zhuo 234S | 931-5 | 4/18 |
Table 5. Tiller ratio of mutagenic M1 mutants
| 基因 Gene | 品种 Variety | 突变株定位(行-株) Location of mutants (row-plant) | 突变分蘖占比 Proportion of tillers containing mutations |
|---|---|---|---|
| OsNRAMP5 | 卓234S Zhuo 234S | 246-8 | 18/18 |
| OsNRAMP5 | 卓234S Zhuo 234S | 234-6 | 4/18 |
| OsNRAMP5 | 湘农恢1126 Xiangnonghui 1126 | 204-6 | 3/6 |
| OsNRAMP5 | 湘农恢1126 Xiangnonghui 1126 | 37-7 | 1/8 |
| OsBADH2 | 卓234S Zhuo 234S | 168-1 | 3/18 |
| OsBADH2 | 湘农恢1126 Xiangnonghui 1126 | 224-8 | 1/6 |
| OsLOX3 | 卓234S Zhuo 234S | 760-7 | 1/18 |
| OsLOX3 | 湘农恢1126 Xiangnonghui 1126 | 115-5 | 2/6 |
| OsPAO5 | 卓234S Zhuo 234S | 549-5 | 1/18 |
| OsPAO5 | 卓234S Zhuo 234S | 1028-4 | 5/18 |
| OsPAO5 | 湘农恢1126 Xiangnonghui 1126 | 84-2 | 1/6 |
| OsSSIIIa | 卓234S Zhuo 234S | 598-7 | 1/18 |
| OsBEIIb | 卓234S Zhuo 234S | 931-5 | 4/18 |
Fig. 2. KASP identification of M2 generation and Sanger sequencing of mutant plants A and B, KASP identification of OsNRAMP5 gene in Zhuo 234S and Xiangnonghui 1126 M2 population and Sanger sequencing of mutants. osnramp5246-8 and osnramp537-7 represent mutant plant of OsNRAMP5 gene in Zhuo 234S and Xiangnonghui 1126, respectively; C and D, KASP identification of OsBADH2 gene in Zhuo 234S and Xiangnonghui 1126 M2 population and Sanger sequencing of mutants; osbadh2168-1and osbadh2224-8 represent mutant plant of OsBADH2 gene in Zhuo234S and Xiang Nonghui 1126, respectively. E-G, KASP identification of OsLOX3, OsPAO5 and OsBEⅡb gene in Zhuo 234S M2 population and Sanger sequencing of mutants. oslox3760-7, ospao5549-5 and osbe2b931-5 represented mutant plant of OsLOX3, OsPAO5 and OsBEⅡb gene of Zhuo 234S, respectively. In panel A, red dots represent heterozygous genotype, blue dots represent homozygous mutant genotype, green dots represent wild genotype. In panels B-G, red dots represent heterozygous genotype, green dots represent homozygous mutant genotype, blue dots represent wild genotype. The dots in the red oval box represent the homozygous/heterozygous genotype of target genes.
Fig. 3. Contents of Cd and Mn in homozygous mutant of OsNRAMP5 and control A-D, Contents of Cd and Mn in homozygous mutant Zhuo 234S-osnramp5246-8 and its original control material; E-H, Contents of Cd and Mn in the homozygous mutant Xiangnonghui 1126-osnramp537-7and its original control material; *, **, and *** represent difference was significant at 0.05, 0.01, and 0.001 levels, respectively (t-test).
| 材料 Material | 株高 Plant height (cm) | 有效分蘖数 No. of effective tillers | 主穗长 Panicle length(cm) | 一次枝梗数 No. of primary branches | 二次枝梗数 No. of secondary branches |
|---|---|---|---|---|---|
| 卓两优1126(对照) Zhuoliangyou 1126(CK) | 124.6±3.3 a | 13.2±1.3 a | 28.9±1.4 a | 18.2±1.3 a | 99.4±6.4 a |
| 镉低积累卓两优1126 Zhuoliangyou 1126 with low cadmium accumulation | 124.8±2.7 a | 13.6±1.1 a | 27.9±0.9 a | 19.0±0.7 a | 104.2±5.7 a |
| 材料 Material | 总粒数 Spikelets per panicle | 实粒数 Grains per panicle | 结实率 Seed setting rate(%) | 千粒重 1000-grain weight(g) | |
| 卓两优1126(对照) Zhuoliangyou 1126(CK) | 347.4±39.1 a | 301.6±28.9 a | 87.0±2.0 a | 22.9±0.4 a | |
| 镉低积累卓两优1126 Zhuoliangyou 1126 with low cadmium accumulation | 360.8±24.1 a | 316.8±19.6 a | 88.0±2.0 a | 22.9±0.5 a |
Table 6. Comparison of agronomic traits of Zhuoliangyou 1126 with low cadmium accumulation and its original control varieties
| 材料 Material | 株高 Plant height (cm) | 有效分蘖数 No. of effective tillers | 主穗长 Panicle length(cm) | 一次枝梗数 No. of primary branches | 二次枝梗数 No. of secondary branches |
|---|---|---|---|---|---|
| 卓两优1126(对照) Zhuoliangyou 1126(CK) | 124.6±3.3 a | 13.2±1.3 a | 28.9±1.4 a | 18.2±1.3 a | 99.4±6.4 a |
| 镉低积累卓两优1126 Zhuoliangyou 1126 with low cadmium accumulation | 124.8±2.7 a | 13.6±1.1 a | 27.9±0.9 a | 19.0±0.7 a | 104.2±5.7 a |
| 材料 Material | 总粒数 Spikelets per panicle | 实粒数 Grains per panicle | 结实率 Seed setting rate(%) | 千粒重 1000-grain weight(g) | |
| 卓两优1126(对照) Zhuoliangyou 1126(CK) | 347.4±39.1 a | 301.6±28.9 a | 87.0±2.0 a | 22.9±0.4 a | |
| 镉低积累卓两优1126 Zhuoliangyou 1126 with low cadmium accumulation | 360.8±24.1 a | 316.8±19.6 a | 88.0±2.0 a | 22.9±0.5 a |
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