Chinese Journal OF Rice Science ›› 2023, Vol. 37 ›› Issue (4): 359-367.DOI: 10.16819/j.1001-7216.2023.221113
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LI Gang1, GAO Qingsong2, LI Wei2, ZHANG Wenxia2, WANG Jian1, CHEN Baoshan1, WANG Di1, GAO Hao1, XU Weijun1, CHEN Hongqi3(), JI Jianhui2()
Received:
2022-11-24
Revised:
2023-02-08
Online:
2023-07-10
Published:
2023-07-17
Contact:
*email: chqhzfy@126.com;jijianhui@hytc.edu.cn
李刚1, 高清松2, 李伟2, 张雯霞2, 王健1, 程保山1, 王迪1, 高浩1, 徐卫军1, 陈红旗3(), 纪剑辉2()
通讯作者:
*email: chqhzfy@126.com;jijianhui@hytc.edu.cn
基金资助:
LI Gang, GAO Qingsong, LI Wei, ZHANG Wenxia, WANG Jian, CHEN Baoshan, WANG Di, GAO Hao, XU Weijun, CHEN Hongqi, JI Jianhui. Directed Knockout of SD1 Gene Improves Lodging Resistance and Blast Resistance of Rice[J]. Chinese Journal OF Rice Science, 2023, 37(4): 359-367.
李刚, 高清松, 李伟, 张雯霞, 王健, 程保山, 王迪, 高浩, 徐卫军, 陈红旗, 纪剑辉. 定向敲除SD1基因提高水稻的抗倒性和稻瘟病抗性[J]. 中国水稻科学, 2023, 37(4): 359-367.
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URL: http://www.ricesci.cn/EN/10.16819/j.1001-7216.2023.221113
引物名称 Primer name | 序列(5’-3’) Primer sequence(5'-3') |
---|---|
SD1-P1 | TGTGGAGCCATTCGTGTGGCCGAA |
SD1-P2 | AAACTTCGGCCACACGAATGGCTC |
SD1-F | CACAGCCGCACCAACCAC |
SD1-R | GTGGAAGCCGAAGGAGAGG |
HPTF | GGGTGTCACGTTGCAAGACC |
HPTR | ATGCCTCCGCTCGAAGTAGC |
sgRNA-F | TCCCAGTCACGACGTTGTAA |
sgRNA-R | GGCCATTTGTCTGCAGAAT |
Cas9-F | CACCATCTACCACCTGAGAA |
Cas9-R | CGAAGTTGCTCTTGAAGTTG |
Actin-F | CCCCTCCTGAAAGGAAGTA |
Actin-R | GGTCCGAAGAATTAGAAGCA |
Table 1. Primer sequences used in this study.
引物名称 Primer name | 序列(5’-3’) Primer sequence(5'-3') |
---|---|
SD1-P1 | TGTGGAGCCATTCGTGTGGCCGAA |
SD1-P2 | AAACTTCGGCCACACGAATGGCTC |
SD1-F | CACAGCCGCACCAACCAC |
SD1-R | GTGGAAGCCGAAGGAGAGG |
HPTF | GGGTGTCACGTTGCAAGACC |
HPTR | ATGCCTCCGCTCGAAGTAGC |
sgRNA-F | TCCCAGTCACGACGTTGTAA |
sgRNA-R | GGCCATTTGTCTGCAGAAT |
Cas9-F | CACCATCTACCACCTGAGAA |
Cas9-R | CGAAGTTGCTCTTGAAGTTG |
Actin-F | CCCCTCCTGAAAGGAAGTA |
Actin-R | GGTCCGAAGAATTAGAAGCA |
Fig. 1. Target site for SD1 gene knock-out design, T-DNA structure and mutation identification. A, SD1 gene structure and gene knockout target sites; B, Off target sequence analysis; C, Gene editing vector T-DNA segment structure; D, Sequencing results and corresponding amino acid sequence.
试验小区 Experiment plot | 倒伏比例 Lodging percentage/% | Yield 产量/(t·hm−2) | ||
---|---|---|---|---|
野生型 Wild type | sd1 | 野生型 Wild type | sd1 | |
重复小区1 Repeat 1 | 60 | 0 | 10.52 | 10.31 |
重复小区2 Repeat 2 | 90 | 0 | 11.26 | 11.53 |
重复小区3 Repeat 3 | 75 | 0 | 11.55 | 11.46 |
平均 Mean | 75 | 0 | 11.11 | 11.10 |
Table 2. Yield and lodging result of the wild-type Huai 119 and sd1 mutant.
试验小区 Experiment plot | 倒伏比例 Lodging percentage/% | Yield 产量/(t·hm−2) | ||
---|---|---|---|---|
野生型 Wild type | sd1 | 野生型 Wild type | sd1 | |
重复小区1 Repeat 1 | 60 | 0 | 10.52 | 10.31 |
重复小区2 Repeat 2 | 90 | 0 | 11.26 | 11.53 |
重复小区3 Repeat 3 | 75 | 0 | 11.55 | 11.46 |
平均 Mean | 75 | 0 | 11.11 | 11.10 |
Fig. 4. Plant height of the wild type and the mutant under the exogenous GA treatment. A, GA treatment for 7 days; B, GA treatment for 10 days; *Significant difference at 0.05 level between the wild type and the mutant (n=4).
品种名称Variety name | 苗瘟病级Disease rating of seedling blast | 叶瘟病级Disease rating of leaf blast | 穗颈瘟Neck blast | 抗性 Resistance | ||||||
---|---|---|---|---|---|---|---|---|---|---|
发病率Incidence rate of blast/% | 病级Disease rating | 损失率 Loss rate of blast /% | 病级 Disease rating | 抗性综合指数Comprehensive resistance index | 病级Disease rating | |||||
sd1 | 0 | 1 | 23 | 5 | 6 | 3 | 3.00 | 3 | 中抗Moderately resistant | |
淮119 Huai 119 | 0 | 2 | 82 | 9 | 33 | 7 | 6.25 | 7 | 感病Sensitive |
Table 3. Resistance evaluation of sd1 and Huai 119 to rice blast.
品种名称Variety name | 苗瘟病级Disease rating of seedling blast | 叶瘟病级Disease rating of leaf blast | 穗颈瘟Neck blast | 抗性 Resistance | ||||||
---|---|---|---|---|---|---|---|---|---|---|
发病率Incidence rate of blast/% | 病级Disease rating | 损失率 Loss rate of blast /% | 病级 Disease rating | 抗性综合指数Comprehensive resistance index | 病级Disease rating | |||||
sd1 | 0 | 1 | 23 | 5 | 6 | 3 | 3.00 | 3 | 中抗Moderately resistant | |
淮119 Huai 119 | 0 | 2 | 82 | 9 | 33 | 7 | 6.25 | 7 | 感病Sensitive |
Fig. 6. Phenotype of the wild type Huai 119 and sd1 mutant under the chlorate treatment. A, Seedlings treated with KClO3 during the germination period, Bar= 2 cm; B, Height of seedlings treated with KClO3 during germination, n=10; C, Seedlings treated with KClO3 at seedling stage, scale=5 cm; D and E, Total chlorophyll (D) and carotenoid (E) contents of seedlings after KClO3 treatment at seedling stage.
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