Chinese Journal OF Rice Science ›› 2021, Vol. 35 ›› Issue (3): 259-268.DOI: 10.16819/j.1001-7216.2021.01204
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Xuemei DENG1,#, Peng HU1,#, Yueying WANG1, Yi WEN1, Yiqing TAN1, Hao WU2, Kaixiong WU1, Junge WANG1, Linlin HOU1, Lixin ZHU1, Li ZHU1, Guang CHEN1, Dali ZENG1, Guangheng ZHANG1, Longbiao GUO1, Zhenyu GAO1, Deyong REN1, Qian QIAN1,*(), Jiang HU1,*()
Received:
2020-06-10
Revised:
2020-08-06
Online:
2021-05-10
Published:
2021-05-10
Contact:
Qian QIAN, Jiang HU
About author:
#These authors contributed equally to this work;
邓雪梅1,#, 胡鹏1,#, 王月影1, 文艺1, 谭义青1, 伍豪2, 吴凯雄1, 王俊格1, 侯琳琳1, 朱黎欣1, 朱丽1, 陈光1, 曾大力1, 张光恒1, 郭龙彪1, 高振宇1, 任德勇1, 钱前1,*(), 胡江1,*()
通讯作者:
钱前,胡江
作者简介:
#共同第一作者;
基金资助:
Xuemei DENG, Peng HU, Yueying WANG, Yi WEN, Yiqing TAN, Hao WU, Kaixiong WU, Junge WANG, Linlin HOU, Lixin ZHU, Li ZHU, Guang CHEN, Dali ZENG, Guangheng ZHANG, Longbiao GUO, Zhenyu GAO, Deyong REN, Qian QIAN, Jiang HU. Identification and Fine Mapping of a Grain Width Mutant gw4 in Rice[J]. Chinese Journal OF Rice Science, 2021, 35(3): 259-268.
邓雪梅, 胡鹏, 王月影, 文艺, 谭义青, 伍豪, 吴凯雄, 王俊格, 侯琳琳, 朱黎欣, 朱丽, 陈光, 曾大力, 张光恒, 郭龙彪, 高振宇, 任德勇, 钱前, 胡江. 水稻粒宽突变体gw4的鉴定与基因定位[J]. 中国水稻科学, 2021, 35(3): 259-268.
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URL: http://www.ricesci.cn/EN/10.16819/j.1001-7216.2021.01204
分子标记 | 正向引物序列(5'-3') | 反向引物序列(5'-3') | 实验目的 | |
---|---|---|---|---|
Marker | Forward primer sequence (5'-3') | Reverse primer sequence (5'-3') | Purpose | |
BS6 | AGTACACATGGGGAGTAGTTGG | GATGGCAATTGAAAAGTGACC | 定位Mapping | |
BS11 | TGCAGGTACACACACATACCC | GGAAGTAGCATCCATGCAACTA | 定位Mapping | |
BS15 | TTTGCAGCTGGCTAGCTTA | ACTTCCATAATTACGGGTCCT | 定位Mapping | |
BS16 | GGCAAATAATTAATAGCATGGTCC | TGGATGATTTCTGTTAGTGGTTCA | 定位Mapping | |
BS18 | TGGTTACAACGACACAACTGC | TCACTCATACCAACCTTGCG | 定位Mapping | |
EX49 | GTGCCTCCTTTTCTTGGAGAT | CACAGTCATCTCACTTGCAATC | 定位Mapping | |
BS19 | ACGTGTCTTGGGGCTGTGTC | TCCCTACACTTATTGCCCCTCT | 测序Sequencing | |
BS20 | TCCTCATTACATGTCCCCATCC | CCCTATCTGAACGCAAGTGCT | 测序Sequencing | |
BS21 | GTGGTAGCATGTGCTCTCCCT | AGGGTTGGCAGAGTTGTCAGA | 测序Sequencing | |
BS22 | TACCATGTTGCACCTGAATACCT | CAACCCTCCTGGTGATTCCC | 测序Sequencing | |
BS23 | CCAGTTGTTAGGGCTGTGTCTG | AGATTAGGGAGCCAATGCGA | 测序Sequencing | |
BS24 | GATGCTCTGTTTGGACAGCCT | TGTCGTCAGCATGTTGCCTG | 测序Sequencing | |
BS25 | ACCAACCAACCCCTTGAGTCT | TGGAAATCGGAGAGATTATACGTAC | 测序Sequencing | |
Histon | GGTCAACTTGTTGATTCCCCTCT | AACCGCAAAATCCAAAGAACG | RT-PCR | |
RT-GW2 | ATCGTGGACAAGGGCACCTGC | CCGTCCCGCTCGCGAGGCATG | RT-PCR | |
RT-GW5 | AATCCAGACGGCATTCAGAG | CTTCACCAGCGCCTTGAG | RT-PCR | |
RT-GW7 | CCCTAGCATCGACACCAAGT | TCGAGGACAGAGATGGGACT | RT-PCR | |
RT-GW8 | ACAGCCAGATCCCATGAACT | GCGTGTAGTATGGGCTCTCC | RT-PCR | |
RT-GS2 | TGCGTCCCTTCTTTGATGAGT | ACAGTTGGGTGCCTGAGAATG | RT-PCR | |
RT-GS3 | CGGAAGAACTCCTGATCCATTC | CACTTGCTCTGCACAAACAGC | RT-PCR | |
RT-GS5 | GTTCTCGGTACTGCGTGGAAG | ACTCCACAAACCTCCCAGCA | RT-PCR | |
RT-GL6 | TCCAAACCAGATGTTGGTGA | GCCGTCGACTACATCTCGTT | RT-PCR | |
RT-LOC_Os04g01590 | TTGGTGATGTCCCCATACAA | ACCAAATCTCTTCCCCTGCT | RT-PCR |
Table 1. Markers developed for mapping of gw4 and quantitative real-time PCR (qRT- PCR) analysis.
分子标记 | 正向引物序列(5'-3') | 反向引物序列(5'-3') | 实验目的 | |
---|---|---|---|---|
Marker | Forward primer sequence (5'-3') | Reverse primer sequence (5'-3') | Purpose | |
BS6 | AGTACACATGGGGAGTAGTTGG | GATGGCAATTGAAAAGTGACC | 定位Mapping | |
BS11 | TGCAGGTACACACACATACCC | GGAAGTAGCATCCATGCAACTA | 定位Mapping | |
BS15 | TTTGCAGCTGGCTAGCTTA | ACTTCCATAATTACGGGTCCT | 定位Mapping | |
BS16 | GGCAAATAATTAATAGCATGGTCC | TGGATGATTTCTGTTAGTGGTTCA | 定位Mapping | |
BS18 | TGGTTACAACGACACAACTGC | TCACTCATACCAACCTTGCG | 定位Mapping | |
EX49 | GTGCCTCCTTTTCTTGGAGAT | CACAGTCATCTCACTTGCAATC | 定位Mapping | |
BS19 | ACGTGTCTTGGGGCTGTGTC | TCCCTACACTTATTGCCCCTCT | 测序Sequencing | |
BS20 | TCCTCATTACATGTCCCCATCC | CCCTATCTGAACGCAAGTGCT | 测序Sequencing | |
BS21 | GTGGTAGCATGTGCTCTCCCT | AGGGTTGGCAGAGTTGTCAGA | 测序Sequencing | |
BS22 | TACCATGTTGCACCTGAATACCT | CAACCCTCCTGGTGATTCCC | 测序Sequencing | |
BS23 | CCAGTTGTTAGGGCTGTGTCTG | AGATTAGGGAGCCAATGCGA | 测序Sequencing | |
BS24 | GATGCTCTGTTTGGACAGCCT | TGTCGTCAGCATGTTGCCTG | 测序Sequencing | |
BS25 | ACCAACCAACCCCTTGAGTCT | TGGAAATCGGAGAGATTATACGTAC | 测序Sequencing | |
Histon | GGTCAACTTGTTGATTCCCCTCT | AACCGCAAAATCCAAAGAACG | RT-PCR | |
RT-GW2 | ATCGTGGACAAGGGCACCTGC | CCGTCCCGCTCGCGAGGCATG | RT-PCR | |
RT-GW5 | AATCCAGACGGCATTCAGAG | CTTCACCAGCGCCTTGAG | RT-PCR | |
RT-GW7 | CCCTAGCATCGACACCAAGT | TCGAGGACAGAGATGGGACT | RT-PCR | |
RT-GW8 | ACAGCCAGATCCCATGAACT | GCGTGTAGTATGGGCTCTCC | RT-PCR | |
RT-GS2 | TGCGTCCCTTCTTTGATGAGT | ACAGTTGGGTGCCTGAGAATG | RT-PCR | |
RT-GS3 | CGGAAGAACTCCTGATCCATTC | CACTTGCTCTGCACAAACAGC | RT-PCR | |
RT-GS5 | GTTCTCGGTACTGCGTGGAAG | ACTCCACAAACCTCCCAGCA | RT-PCR | |
RT-GL6 | TCCAAACCAGATGTTGGTGA | GCCGTCGACTACATCTCGTT | RT-PCR | |
RT-LOC_Os04g01590 | TTGGTGATGTCCCCATACAA | ACCAAATCTCTTCCCCTGCT | RT-PCR |
Fig. 1. Grains size of the wild type Zhonghua 11(ZH11) and its mutant gw4. A and B, Mature grain of WT and gw4, bar= 0.25 cm; C-F, Bars represent standard error(n=13). **indicate significant difference between WT and gw4 by t-test(P<0.01). ZH11, Zhonghua 11.
Fig. 2. Phenotype of the wild type Zhonghua 11(ZH11) and its mutant gw4. A and B, Phenotype of Zhonghua 11(ZH11) and gw4, bar=20 cm; C, Internodes, bar=5 cm; D, E and F, Panicles, bar=5 cm; G-L, Bars represent standard error (n=10). ** and * indicate significant difference between WT and gw4 by t-test(P<0.01, P<0.05), respectively.
Fig. 3. Glumes of the wild type(ZH11) and its mutant gw4. A, Glume, bar=0.25 cm; B and C, Outer epidermis of glume, bar=50 μm; D and E, Inner epidermis of glume; bar=50 μm; Bars represent standard error (n=10). **indicate significant difference between WT and gw4 by t-test(P<0.01).
Fig. 4. Fine mapping of gw4. A, gw4 was primarily mapped on chromosome 4; B, gw4 was narrowed to a 31.74 kb genomic region; C, Total six genes was predicted in this region.
Fig. 5. Three-dimensional structures of proteins GW4 and gw4. A and B, Protein domain of GW4; C and D, Protein domain of gw4; B and D, Enlarged size of A and C; The red box indicate the differences in protein folding.
Fig. 6. Expression analysis of LOC_Os04g01590 and grain shape genes. A, Expression analysis of LOC_Os04g01590 in Zhonghua 11(ZH11) and gw4; B, Relative transcript levels of LOC_Os04g01590 in various organs; C, Expression analysis of grain shape genes in the ZH11 and gw4. Set the transcription level in the panicle of ZH11 to 1.0. Bars represent standard deviation (n=3). ** indicates significant difference between ZH11 and gw4 by t-test (P<0.01).
基因 Gene | 功能注释 Function annotation |
---|---|
LOC_Os04g01560 | 表达蛋白Expressed protein |
LOC_Os04g01570 | 蔗糖酶/果胶甲基酯酶抑制剂家族蛋白Invertase/Pectin methylesterase inhibitor family protein |
LOC_Os04g01580 | 表达蛋白Expressed protein |
LOC_Os04g01590 | 精氨酸酶Arginase |
LOC_Os04g01600 | 赤藓酸-4-磷酸脱氢酶结构域蛋白Erythroate-4-phosphate dehydrogenase domain protein |
LOC_Os04g01610 | 表达蛋白Expressed protein |
Table 2 Predicted genes in fine-mapping region.
基因 Gene | 功能注释 Function annotation |
---|---|
LOC_Os04g01560 | 表达蛋白Expressed protein |
LOC_Os04g01570 | 蔗糖酶/果胶甲基酯酶抑制剂家族蛋白Invertase/Pectin methylesterase inhibitor family protein |
LOC_Os04g01580 | 表达蛋白Expressed protein |
LOC_Os04g01590 | 精氨酸酶Arginase |
LOC_Os04g01600 | 赤藓酸-4-磷酸脱氢酶结构域蛋白Erythroate-4-phosphate dehydrogenase domain protein |
LOC_Os04g01610 | 表达蛋白Expressed protein |
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