植物学报 ›› 2023, Vol. 58 ›› Issue (3): 394-403.DOI: 10.11983/CBB22259
金佳怡1, 罗怿婷1, 杨惠敏1, 芦涛1, 叶涵斐1, 谢继毅1, 王珂欣1, 陈芊羽1, 方媛1, 王跃星2(), 饶玉春1()
收稿日期:
2022-11-08
接受日期:
2023-02-09
出版日期:
2023-05-01
发布日期:
2023-05-17
通讯作者:
*E-mail: wangyuexing@caas.cn; ryc@zjnu.cn
基金资助:
Jiayi Jin1, Yiting Luo1, Huimin Yang1, Tao Lu1, Hanfei Ye1, Jiyi Xie1, Kexin Wang1, Qianyu Chen1, Yuan Fang1, Yuexing Wang2(), Yuchun Rao1()
Received:
2022-11-08
Accepted:
2023-02-09
Online:
2023-05-01
Published:
2023-05-17
Contact:
*E-mail: wangyuexing@caas.cn; ryc@zjnu.cn
摘要: 水稻(Oryza sativa)是我国主要粮食作物之一。提高水稻叶片叶绿素含量, 进而提高其光合作用效率是实现高产稳产的重要途径之一。该研究以父系籼稻品种华占(HZ)、母系粳稻品种热研2号(Nekken2)及其构建的120个重组自交系(RILs)为实验材料, 在分蘖期和成熟期分别对亲本及其后代剑叶的叶绿素含量(SPAD值)进行测定, 同时基于已构建的高密度遗传连锁图谱进行QTL定位。结果共挖掘到20个与叶绿素含量相关的QTLs (分蘖期7个, 成熟期13个), LOD值最高达4.77。利用qRT-PCR方法检测QTL区间内与叶绿素含量相关的候选基因的表达, 发现LOC_Os06g11780、LOC_Os06g12360、LOC_Os06g39716、LOC_Os08g42610、LOC_Os02g18500、LOC_Os03g21240、LOC_Os03g21400、LOC_Os03g21780、LOC_Os03g30950及LOC_Os03g40550基因的表达量在双亲间差异显著。结合基因表达量及亲本叶绿素表型数据, 推测LOC_Os06g11780、LOC_Os06g12360和LOC_Os08g42610的高表达极大地提高了水稻叶绿素含量, 进而有效提高植物光合产能。研究结果为筛选和培育高光能利用效率的水稻新品种提供了有利的遗传资源, 并为揭示水稻叶绿素含量的动态变化规律及分子调控机制奠定了重要基础。
金佳怡, 罗怿婷, 杨惠敏, 芦涛, 叶涵斐, 谢继毅, 王珂欣, 陈芊羽, 方媛, 王跃星, 饶玉春. 水稻叶绿素含量QTL定位与候选基因表达分析. 植物学报, 2023, 58(3): 394-403.
Jiayi Jin, Yiting Luo, Huimin Yang, Tao Lu, Hanfei Ye, Jiyi Xie, Kexin Wang, Qianyu Chen, Yuan Fang, Yuexing Wang, Yuchun Rao. QTL Mapping and Expression Analysis on Candidate Genes Related to Chlorophyll Content in Rice. Chinese Bulletin of Botany, 2023, 58(3): 394-403.
Primer name | Sequence (5′-3′) |
---|---|
LOC_Os06g11780-F-qrt | TGTTGGTCCCTGAAAACTCC |
LOC_Os06g11780-R-qrt | TCAAGCAAATTCACCCATGA |
LOC_Os06g12360-F-qrt | ATGCAAAAAGGAGAGGCTGA |
LOC_Os06g12360-R-qrt | ATGCCGATCAGGAATCTCAC |
LOC_Os06g39650-F-qrt | GTGCTCTTCAACCAGTGCAA |
LOC_Os06g39650-R-qrt | AGATTGGCACAGCATGTCAG |
LOC_Os06g39716-F-qrt | CAACCTTTTCGTAGGCATCC |
LOC_Os06g39716-R-qrt | TTGAAATTGACTGGGCTTGA |
LOC_Os08g42610-F-qrt | GCTCGCACACTGAAAACAAA |
LOC_Os08g42610-R-qrt | ATGGGATAGGCCCCAAATAG |
LOC_Os02g18500-F-qrt | AGGAGGAGGTGTTGGGAACT |
LOC_Os02g18500-R-qrt | ATGCAGTTGTTGATGGCGTA |
LOC_Os03g21240-F-qrt | TATGCCTTTGCACCAGAGTG |
LOC_Os03g21240-R-qrt | CAGCATTCGACGACTGAGAA |
LOC_Os03g21400-F-qrt | AGGAAAGCGGAGAAGAAAGG |
LOC_Os03g21400-R-qrt | TTAGACACCAGGCAGCTGTG |
LOC_Os03g21780-F-qrt | CTCCCTTCCACTCCTCCTTC |
LOC_Os03g21780-R-qrt | CTTCTCCTCCTCGACACCAG |
LOC_Os03g30950-F-qrt | TCTTCTTCTCCATGGCCTGT |
LOC_Os03g30950-R-qrt | AGCAGGGAAGGAAGTCGTG |
LOC_Os03g40550-F-qrt | CTTTGCTCCCTGCCTTGTAG |
LOC_Os03g40550-R-qrt | CACGTCTCGGAGCTCTTTTC |
表1 qRT-PCR所用引物序列
Table 1 Primer sequences for qRT-PCR
Primer name | Sequence (5′-3′) |
---|---|
LOC_Os06g11780-F-qrt | TGTTGGTCCCTGAAAACTCC |
LOC_Os06g11780-R-qrt | TCAAGCAAATTCACCCATGA |
LOC_Os06g12360-F-qrt | ATGCAAAAAGGAGAGGCTGA |
LOC_Os06g12360-R-qrt | ATGCCGATCAGGAATCTCAC |
LOC_Os06g39650-F-qrt | GTGCTCTTCAACCAGTGCAA |
LOC_Os06g39650-R-qrt | AGATTGGCACAGCATGTCAG |
LOC_Os06g39716-F-qrt | CAACCTTTTCGTAGGCATCC |
LOC_Os06g39716-R-qrt | TTGAAATTGACTGGGCTTGA |
LOC_Os08g42610-F-qrt | GCTCGCACACTGAAAACAAA |
LOC_Os08g42610-R-qrt | ATGGGATAGGCCCCAAATAG |
LOC_Os02g18500-F-qrt | AGGAGGAGGTGTTGGGAACT |
LOC_Os02g18500-R-qrt | ATGCAGTTGTTGATGGCGTA |
LOC_Os03g21240-F-qrt | TATGCCTTTGCACCAGAGTG |
LOC_Os03g21240-R-qrt | CAGCATTCGACGACTGAGAA |
LOC_Os03g21400-F-qrt | AGGAAAGCGGAGAAGAAAGG |
LOC_Os03g21400-R-qrt | TTAGACACCAGGCAGCTGTG |
LOC_Os03g21780-F-qrt | CTCCCTTCCACTCCTCCTTC |
LOC_Os03g21780-R-qrt | CTTCTCCTCCTCGACACCAG |
LOC_Os03g30950-F-qrt | TCTTCTTCTCCATGGCCTGT |
LOC_Os03g30950-R-qrt | AGCAGGGAAGGAAGTCGTG |
LOC_Os03g40550-F-qrt | CTTTGCTCCCTGCCTTGTAG |
LOC_Os03g40550-R-qrt | CACGTCTCGGAGCTCTTTTC |
图1 水稻重组自交系中叶绿素含量分布 (A) 分蘖期; (B) 成熟期
Figure 1 Distribution of chlorophyll content in recombinant inbred lines of rice (A) Tillering stage; (B) Maturing stage
图2 父本华占与母本热研2号的SPAD值及表型 (A) 分蘖期水稻植株的SPAD值; (B) 分蘖期水稻植株的表型(bar=10 cm); (C) 成熟期水稻植株的SPAD值; (D) 成熟期水稻植株的表型(bar=10 cm)
Figure 2 SPAD value and phenotype of the male parent HZ and the female parent Nekken2 (A) SPAD value of rice plant at tillering stage; (B) Phenotype of rice plant at tillering stage (bar=10 cm); (C) SPAD value of rice plant at maturing stage; (D) Phenotype of rice plant at maturing stage (bar=10 cm)
Trait | QTL | Chromosome | Physical distance (bp) | Position of support (cM) | Likelihood of odd (LOD) |
---|---|---|---|---|---|
Chlorophyll content at tillering stage | qFLCC6.1 | 6 | 6015262-6720585 | 25.79-28.81 | 3.13 |
qFLCC6.2 | 6 | 22337450-23745958 | 95.75-101.79 | 2.92 | |
qFLCC6.3 | 6 | 24051716-24198880 | 103.10-103.73 | 2.13 | |
qFLCC6.4 | 6 | 25991018-26126806 | 111.42-111.99 | 2.12 | |
qFLCC8.1 | 8 | 26687999-27132649 | 114.40-116.31 | 2.50 | |
qFLCC8.2 | 8 | 27513012-27730624 | 117.94-118.87 | 2.28 | |
qFLCC9.1 | 9 | 20982305-21030827 | 89.95-90.15 | 2.09 | |
Chlorophyll content at maturing stage | qFLCC1.1 | 1 | 2407127-2649258 | 10.32-11.36 | 2.63 |
qFLCC2.1 | 2 | 10729809-10871055 | 45.99-46.60 | 2.01 | |
qFLCC3.1 | 3 | 8989609-9140871 | 38.54-39.18 | 2.00 | |
qFLCC3.2 | 3 | 11602077-12505077 | 49.73-53.61 | 4.77 | |
qFLCC3.3 | 3 | 17425033-18170152 | 74.70-77.90 | 2.38 | |
qFLCC3.4 | 3 | 19142858-20859360 | 82.06-89.42 | 2.29 | |
qFLCC3.5 | 3 | 22349012-22880743 | 95.80-98.08 | 2.59 | |
qFLCC3.6 | 3 | 23094070-24507431 | 98.99-105.06 | 3.25 | |
qFLCC4.1 | 4 | 867957-1087636 | 3.72-4.66 | 2.40 | |
qFLCC4.2 | 4 | 2647701-3012559 | 11.35-12.91 | 2.30 | |
qFLCC4.3 | 4 | 6230638-6599549 | 26.71-28.29 | 2.46 | |
qFLCC7.1 | 7 | 9758981-9899583 | 41.83-42.68 | 2.45 | |
qFLCC11.1 | 11 | 3100542-3229790 | 13.29-13.85 | 2.27 |
表2 水稻重组自交系群体叶绿素含量的QTL分析
Table 2 QTL analysis of chlorophyll content of rice recombinant inbred lines
Trait | QTL | Chromosome | Physical distance (bp) | Position of support (cM) | Likelihood of odd (LOD) |
---|---|---|---|---|---|
Chlorophyll content at tillering stage | qFLCC6.1 | 6 | 6015262-6720585 | 25.79-28.81 | 3.13 |
qFLCC6.2 | 6 | 22337450-23745958 | 95.75-101.79 | 2.92 | |
qFLCC6.3 | 6 | 24051716-24198880 | 103.10-103.73 | 2.13 | |
qFLCC6.4 | 6 | 25991018-26126806 | 111.42-111.99 | 2.12 | |
qFLCC8.1 | 8 | 26687999-27132649 | 114.40-116.31 | 2.50 | |
qFLCC8.2 | 8 | 27513012-27730624 | 117.94-118.87 | 2.28 | |
qFLCC9.1 | 9 | 20982305-21030827 | 89.95-90.15 | 2.09 | |
Chlorophyll content at maturing stage | qFLCC1.1 | 1 | 2407127-2649258 | 10.32-11.36 | 2.63 |
qFLCC2.1 | 2 | 10729809-10871055 | 45.99-46.60 | 2.01 | |
qFLCC3.1 | 3 | 8989609-9140871 | 38.54-39.18 | 2.00 | |
qFLCC3.2 | 3 | 11602077-12505077 | 49.73-53.61 | 4.77 | |
qFLCC3.3 | 3 | 17425033-18170152 | 74.70-77.90 | 2.38 | |
qFLCC3.4 | 3 | 19142858-20859360 | 82.06-89.42 | 2.29 | |
qFLCC3.5 | 3 | 22349012-22880743 | 95.80-98.08 | 2.59 | |
qFLCC3.6 | 3 | 23094070-24507431 | 98.99-105.06 | 3.25 | |
qFLCC4.1 | 4 | 867957-1087636 | 3.72-4.66 | 2.40 | |
qFLCC4.2 | 4 | 2647701-3012559 | 11.35-12.91 | 2.30 | |
qFLCC4.3 | 4 | 6230638-6599549 | 26.71-28.29 | 2.46 | |
qFLCC7.1 | 7 | 9758981-9899583 | 41.83-42.68 | 2.45 | |
qFLCC11.1 | 11 | 3100542-3229790 | 13.29-13.85 | 2.27 |
Chromosome | QTL locus | Gene ID | Functional annotation | Reference |
---|---|---|---|---|
6 | qFLCC6.1 | LOC_Os06g11780 | MYB family transcription factor, putative, expressed | |
6 | qFLCC6.1 | LOC_Os06g12360 | Pentatricopeptide, putative, expressed | |
6 | qFLCC6.2 | LOC_Os06g39650 | Pentatricopeptide repeat protein | Jiang et al., |
6 | qFLCC6.2 | LOC_Os06g39716 | Chloroplast 30S ribosomal protein S18, putative, expressed | |
8 | qFLCC8.1 | LOC_Os08g42610 | Pentatricopeptide, putative, expressed |
表3 水稻分蘖期叶绿素含量相关候选基因的功能
Table 3 The function of candidate genes related to chlorophyll content of rice at tillering stage
Chromosome | QTL locus | Gene ID | Functional annotation | Reference |
---|---|---|---|---|
6 | qFLCC6.1 | LOC_Os06g11780 | MYB family transcription factor, putative, expressed | |
6 | qFLCC6.1 | LOC_Os06g12360 | Pentatricopeptide, putative, expressed | |
6 | qFLCC6.2 | LOC_Os06g39650 | Pentatricopeptide repeat protein | Jiang et al., |
6 | qFLCC6.2 | LOC_Os06g39716 | Chloroplast 30S ribosomal protein S18, putative, expressed | |
8 | qFLCC8.1 | LOC_Os08g42610 | Pentatricopeptide, putative, expressed |
Chromosome | QTL locus | Gene ID | Functional annotation | Reference |
---|---|---|---|---|
2 | qFLCC2.1 | LOC_Os02g18500 | Chlorophyllase-2, chloroplast precursor, putative | |
3 | qFLCC3.2 | LOC_Os03g21240 | Transcription factor with a MYB domain; PHOSPHATE RESPONSE 1 | Guo et al., |
3 | qFLCC3.2 | LOC_Os03g21400 | Cytochrome P450 gene; gibberellin 13-oxidase | |
3 | qFLCC3.2 | LOC_Os03g21780 | Pentatricopeptide repeat domain containing protein, putative, expressed | |
3 | qFLCC3.3 | LOC_Os03g30950 | Acyl-desaturase, chloroplast precursor, putative, expressed | |
3 | qFLCC3.5 | LOC_Os03g40550 | pfkB-type carbohydrate kinase family protein |
表4 水稻成熟期叶绿素含量相关候选基因的功能
Table 4 The function of candidate genes related to chlorophyll content of rice at maturing stage
Chromosome | QTL locus | Gene ID | Functional annotation | Reference |
---|---|---|---|---|
2 | qFLCC2.1 | LOC_Os02g18500 | Chlorophyllase-2, chloroplast precursor, putative | |
3 | qFLCC3.2 | LOC_Os03g21240 | Transcription factor with a MYB domain; PHOSPHATE RESPONSE 1 | Guo et al., |
3 | qFLCC3.2 | LOC_Os03g21400 | Cytochrome P450 gene; gibberellin 13-oxidase | |
3 | qFLCC3.2 | LOC_Os03g21780 | Pentatricopeptide repeat domain containing protein, putative, expressed | |
3 | qFLCC3.3 | LOC_Os03g30950 | Acyl-desaturase, chloroplast precursor, putative, expressed | |
3 | qFLCC3.5 | LOC_Os03g40550 | pfkB-type carbohydrate kinase family protein |
图4 水稻叶绿素含量相关基因表达量差异 (A) 分蘖期叶绿素含量相关基因表达量; (B) 成熟期叶绿素含量相关基因表达量。**表示在0.01水平差异显著。
Figure 4 Differences in the expression level of the genes related to chlorophyll content in rice (A) Expression of genes related to chlorophyll content at tillering stage; (B) Expression of genes related to chlorophyll content at maturing stage. ** indicate significant differences at 0.01 level.
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