Chinese Journal OF Rice Science ›› 2023, Vol. 37 ›› Issue (4): 337-346.DOI: 10.16819/j.1001-7216.2023.221004
• Research Papers • Next Articles
REN Zhiqi1,2, XUE Kexin2,3, DONG Zheng2,3, LI Xiaoxiang2,3, LI Yongzhao2,3, GUO Yujing1,2, LIU Wenqiang2,3, GUO Liang2,3, SHENG Xinnian2,3, LIU Zhixi2,3, PAN Xiaowu1,2,3()
Received:
2022-10-20
Revised:
2022-11-22
Online:
2023-07-10
Published:
2023-07-17
Contact:
*email: pxw137@163.com
任志奇1,2, 薛可欣2,3, 董铮2,3, 李小湘2,3, 黎用朝2,3, 郭玉静1,2, 刘文强2,3, 郭梁2,3, 盛新年2,3, 刘之熙2,3, 潘孝武1,2,3()
通讯作者:
*email: pxw137@163.com
基金资助:
REN Zhiqi, XUE Kexin, DONG Zheng, LI Xiaoxiang, LI Yongzhao, GUO Yujing, LIU Wenqiang, GUO Liang, SHENG Xinnian, LIU Zhixi, PAN Xiaowu. Identification and Gene Mapping of Outcurved Leaf Mutant ocl1 in Rice[J]. Chinese Journal OF Rice Science, 2023, 37(4): 337-346.
任志奇, 薛可欣, 董铮, 李小湘, 黎用朝, 郭玉静, 刘文强, 郭梁, 盛新年, 刘之熙, 潘孝武. 水稻外卷叶突变体ocl1的鉴定及基因定位[J]. 中国水稻科学, 2023, 37(4): 337-346.
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URL: http://www.ricesci.cn/EN/10.16819/j.1001-7216.2023.221004
引物名称 Primer | 用途 Usage | 正向引物(5'→3') Forward primer (5'→3') | 反向引物(5'→3') Reverse primer (5'→3') |
---|---|---|---|
RM217 | 基因定位Gene mapping | ATCGCAGCAATGCCTCGT | GGGTGTGAACAAAGACAC |
ID02356 | 基因定位Gene mapping | CTCACGTAGGTCTTGAGGAG | AGAAGAGGGCAGGAGGAG |
RM19570 | 基因定位Gene mapping | CCCAGATATTCTGTGTGATCATGAGG | GAGTGAATGTGAGCCGTCTATTGG |
RM19575 | 基因定位Gene mapping | TCATCACAAGCTCGTAATCAGG | CCAGAGAATAAGAGGACATGACG |
ID02612 | 基因定位Gene mapping | GCAGTTAATTATTCCATGCG | TTTGAACTCTCCCATATTCG |
ID03100 | 基因定位Gene mapping | CCATGGATGACTCTCTCTCT | ACACCTCCACTCCTCCAT |
RM276 | 基因定位Gene mapping | CTCAACGTTGACACCTCGTG | TCCTCCATCGAGCAGTATCA |
cDNA-S | 测序Sequencing | AGAGAGGAAGAGGCAGAGGTAG | AGAAGATGCTGTCGTAGGTGTC |
OsActin | 内参Reference gene | CATTGGTGCTGAGCGTTTCC | AGAAACAAGCAGGAGGACGG |
RL14 | qRT-PCR | CTCTTTCAGGCATTCCATTGATG | CAACACCTTGTCAGCTTTCAAGC |
ZHD1 | qRT-PCR | CGAGAACGAATGCTCTCTCAG | CGGACCCCGGTATGGTAG |
SLL1 | qRT-PCR | GCCTCTGTGATTGCCATCTAAT | CAGGTGTCCAACCATGAGC |
ROC5 | qRT-PCR | CGCAAGAGGAAGAAGCGATAC | GCTCCAGTTGCGTCTTCATC |
SRL1 | qRT-PCR | TCTCCTGCCTCCTCTGTGTG | TAGGAGGGGTGGTGTTGAAG |
NRL1 | qRT-PCR | TCAGTAGTGTAGTGGTGTCGAGTTCA | GCACTCCTTCATGTGAGCTTCA |
LAC17 | qRT-PCR | CTGCAGATTTGGCACTCGAGAACGTC | CATGCTCTTGGTGTTGCACAG |
EXPA2 | qRT-PCR | GGGCACTCCTACTTCAACCT | TAGGAGTTGCTCTGCCAGTT |
EXPA4 | qRT-PCR | GGGCACTCCTACTTCAACCT | CTGGAAGGAGAGGCTCTGG |
XTH11 | qRT-PCR | ACCTTCTACTTGTCGTCGCA | TGCTGTGGGTTCCAGATGAT |
CESA2 | qRT-PCR | GGTATCCTTGAGATGAGGTGG | GCCTTTGAGGTGACAGTGAA |
CESA3 | qRT-PCR | AAGTTCTTCGGTGGGCTCT | TTTCCAGGATGCCAGTAGC |
Table 1. Primers for gene mapping, sequencing and expression analysis.
引物名称 Primer | 用途 Usage | 正向引物(5'→3') Forward primer (5'→3') | 反向引物(5'→3') Reverse primer (5'→3') |
---|---|---|---|
RM217 | 基因定位Gene mapping | ATCGCAGCAATGCCTCGT | GGGTGTGAACAAAGACAC |
ID02356 | 基因定位Gene mapping | CTCACGTAGGTCTTGAGGAG | AGAAGAGGGCAGGAGGAG |
RM19570 | 基因定位Gene mapping | CCCAGATATTCTGTGTGATCATGAGG | GAGTGAATGTGAGCCGTCTATTGG |
RM19575 | 基因定位Gene mapping | TCATCACAAGCTCGTAATCAGG | CCAGAGAATAAGAGGACATGACG |
ID02612 | 基因定位Gene mapping | GCAGTTAATTATTCCATGCG | TTTGAACTCTCCCATATTCG |
ID03100 | 基因定位Gene mapping | CCATGGATGACTCTCTCTCT | ACACCTCCACTCCTCCAT |
RM276 | 基因定位Gene mapping | CTCAACGTTGACACCTCGTG | TCCTCCATCGAGCAGTATCA |
cDNA-S | 测序Sequencing | AGAGAGGAAGAGGCAGAGGTAG | AGAAGATGCTGTCGTAGGTGTC |
OsActin | 内参Reference gene | CATTGGTGCTGAGCGTTTCC | AGAAACAAGCAGGAGGACGG |
RL14 | qRT-PCR | CTCTTTCAGGCATTCCATTGATG | CAACACCTTGTCAGCTTTCAAGC |
ZHD1 | qRT-PCR | CGAGAACGAATGCTCTCTCAG | CGGACCCCGGTATGGTAG |
SLL1 | qRT-PCR | GCCTCTGTGATTGCCATCTAAT | CAGGTGTCCAACCATGAGC |
ROC5 | qRT-PCR | CGCAAGAGGAAGAAGCGATAC | GCTCCAGTTGCGTCTTCATC |
SRL1 | qRT-PCR | TCTCCTGCCTCCTCTGTGTG | TAGGAGGGGTGGTGTTGAAG |
NRL1 | qRT-PCR | TCAGTAGTGTAGTGGTGTCGAGTTCA | GCACTCCTTCATGTGAGCTTCA |
LAC17 | qRT-PCR | CTGCAGATTTGGCACTCGAGAACGTC | CATGCTCTTGGTGTTGCACAG |
EXPA2 | qRT-PCR | GGGCACTCCTACTTCAACCT | TAGGAGTTGCTCTGCCAGTT |
EXPA4 | qRT-PCR | GGGCACTCCTACTTCAACCT | CTGGAAGGAGAGGCTCTGG |
XTH11 | qRT-PCR | ACCTTCTACTTGTCGTCGCA | TGCTGTGGGTTCCAGATGAT |
CESA2 | qRT-PCR | GGTATCCTTGAGATGAGGTGG | GCCTTTGAGGTGACAGTGAA |
CESA3 | qRT-PCR | AAGTTCTTCGGTGGGCTCT | TTTCCAGGATGCCAGTAGC |
Fig. 1. Plant architecture of the wild type and ocl1. A, Plant architecture of WT and ocl1 at the seedling stage, bar=5 cm; B, Plant architecture of WT and ocl1 at the tillering stage, bar=15 cm; C, Leaf morphology of WT and ocl1 at the tillering stage, bar=1 cm; D, Leaf rolling index of WT and ocl1 at the tillering stage; E, Plant architecture of WT and ocl1 at the heading stage, bar=10 cm; F, Seed setting of WT and ocl1, bar=2 cm.
Fig. 2. Cross-sectional and leaf slice observation of WT and ocl1. A, Cross-sectional observation of WT and ocl1, bar=1 mm; B, Leaf slice observation of WT and ocl1, black lines point to bulliform cells of WT and ocl1, bar=100 μm; C, Number of bulliform cells of WT and ocl1; D, Area of bulliform cells of WT and ocl1; NS means there is no significant difference between the mutant and WT; ** Significant difference between the mutant and WT at 0.01 level. ad, Adaxial side; ab, Abaxial side.
材料 Material | 单株穗数 | 每穗总粒数 | 结实率 | 千粒重 | 单株产量 | |
---|---|---|---|---|---|---|
No. of panicles per plant | No. of spikelets per panicle | Seed-setting rate / % | 1000-grain weight / g | Yield per plant / g | ||
WT | 11.9±0.4 | 94.2±1.3 | 74.4±0.8 | 29.6±0.1 | 24.7±1.1 | |
ocl1 | 11.3±0.1 | 92.4±4.0 | 45.7±2.0** | 24.8±0.1** | 11.8±0.7** |
Table 2. Agronomic traits of the WT and ocl1.
材料 Material | 单株穗数 | 每穗总粒数 | 结实率 | 千粒重 | 单株产量 | |
---|---|---|---|---|---|---|
No. of panicles per plant | No. of spikelets per panicle | Seed-setting rate / % | 1000-grain weight / g | Yield per plant / g | ||
WT | 11.9±0.4 | 94.2±1.3 | 74.4±0.8 | 29.6±0.1 | 24.7±1.1 | |
ocl1 | 11.3±0.1 | 92.4±4.0 | 45.7±2.0** | 24.8±0.1** | 11.8±0.7** |
Fig. 3. Analysis of grain morphology between WT and ocl1. A, Grain length of WT and ocl1, bar=1 cm; B, Grain width of WT and ocl1, bar=5 mm; C, Grain plumpness of WT and ocl1, bar=1 cm; D, Statistical analysis of grain length, grain width and grain thickness of WT and ocl1; **Significant difference between the mutant and WT at 0.01 level.
Fig. 4. Map-based cloning of OCL1. A, Mapping of OCL1 gene. Black boxes, white boxes and black lines represent exons, UTR and introns, respectively. Black arrow indicates the mutation site; B, Sequence comparison of WT and ocl1 at the mutation site; C, Electrophoresis detection of cDNA amplification products; D, cDNA sequence alignment of the OCL1 gene between WT and ocl1; E, Alignment of the amino acid sequence between WT and ocl1.
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