植物学报 ›› 2021, Vol. 56 ›› Issue (3): 275-283.DOI: 10.11983/CBB21039
潘晨阳, 张月, 林晗, 陈芊羽, 杨凯如, 姜嘉骥, 李梦佳, 芦涛, 王珂欣, 路梅, 王盛, 叶涵斐, 饶玉春*(), 胡海涛*(
)
收稿日期:
2021-02-22
接受日期:
2021-03-26
出版日期:
2021-05-01
发布日期:
2021-04-30
通讯作者:
饶玉春,胡海涛
作者简介:
haitao-hu@zjnu.cn基金资助:
Chenyang Pan, Yue Zhang, Han Lin, Qianyu Chen, Kairu Yang, Jiaji Jiang, Mengjia Li, Tao Lu, Kexin Wang, Mei Lu, Sheng Wang, Hanfei Ye, Yuchun Rao*(), Haitao Hu*(
)
Received:
2021-02-22
Accepted:
2021-03-26
Online:
2021-05-01
Published:
2021-04-30
Contact:
Yuchun Rao,Haitao Hu
摘要:
为探究叶片水势(LWP)相关基因在水稻(Oryza sativa)抗旱中的作用及其遗传机制, 以热研2号(Nekken2)和华占(HZ)为亲本以及构建的120个重组自交系(RILs)群体为实验材料, 对水稻分蘖期叶片水势进行检测, 并利用前期基于高通量测序构建的分子遗传连锁图谱进行数量性状基因座(QTL)分析。结果表明, 共检测到5个与水稻分蘖期叶片水势相关的QTLs, 分别位于第2、3、4、11和12号染色体上, LOD值均达2.5以上, 其中位于4号染色体物理距离24 066 261- 30 847 136 bp内QTL的LOD值高达5.15。对这些QTL区间内与水势相关的候选基因进行定量分析, 发现LOC_Os02g56630、LOC_Os02g57720、LOC_Os02g57580、LOC_Os04g43730、LOC_Os04g46490、LOC_Os04g44570和LOC_Os04g44060这7个基因在双亲间表达量差异显著。位于4号染色体QTL区间内LOC_Os04g46490基因的表达在两亲本间存在显著差异。对基因LOC_Os04g46490进行测序分析, 发现该基因在两亲本间共存在6处差异, 从而导致氨基酸序列的改变。通过QTL挖掘及相关基因表达分析, 发现这些基因与水稻叶片水势调控相关, 可能间接影响水稻的抗旱性。检测到的QTL位点对水势相关基因精细定位和克隆具有重要参考价值, 有助于进一步理解水稻叶片水势的遗传基础, 并为培育耐旱水稻新品种提供有利的基因资源。
潘晨阳, 张月, 林晗, 陈芊羽, 杨凯如, 姜嘉骥, 李梦佳, 芦涛, 王珂欣, 路梅, 王盛, 叶涵斐, 饶玉春, 胡海涛. 水稻叶片水势的QTL定位与候选基因分析. 植物学报, 2021, 56(3): 275-283.
Chenyang Pan, Yue Zhang, Han Lin, Qianyu Chen, Kairu Yang, Jiaji Jiang, Mengjia Li, Tao Lu, Kexin Wang, Mei Lu, Sheng Wang, Hanfei Ye, Yuchun Rao, Haitao Hu. QTL Mapping and Candidate Gene Analysis on Rice Leaf Water Potential. Chinese Bulletin of Botany, 2021, 56(3): 275-283.
Primer name | Sequence (5′-3′) | Tm (°C) | Length (bp) |
---|---|---|---|
LOC_Os02g56630-F-qrt | GATGCTACGGAGACTGCACA | 60.02 | 182 |
LOC_Os02g56630-R-qrt | GAGCCAGAAGCACATGAACA | 59.99 | |
LOC_Os02g57720-F-qrt | CTGACGTCGTGGTCGTTCTA | 59.90 | 101 |
LOC_Os02g57720-R-qrt | GACTTGTTCACCCCCATCAC | 60.22 | |
LOC_Os04g43730-F-qrt | ACCAAAGTGGGTGTTTCTCG | 60.01 | 153 |
LOC_Os04g43730-R-qrt | TCGAGATCTGGCTTGTGTTG | 59.98 | |
LOC_Os04g44060-F-qrt | CGGTGTTCATGGTTCACTTG | 60.00 | 182 |
LOC_Os04g44060-R-qrt | CCTCAGGACGTACTGGTGGT | 60.03 | |
LOC_Os04g46490-F-qrt | GCCTCGTCCTCCACTACATC | 59.69 | 123 |
LOC_Os04g46490-R-qrt | CCGTGTACACCACCATGAAC | 59.73 | |
LOC_Os02g57580-F-qrt | ACCTCTTCAGATGGGGTGTG | 59.96 | 101 |
LOC_Os02g57580-R-qrt | CCAGTCAGTTTTGCAGACCA | 59.87 | |
LOC_Os04g48230-F-qrt | GGGCACTACAAGTCCGTGAT | 60.00 | 199 |
LOC_Os04g48230-R-qrt | CTTGGTAGCTTCCGATGAGC | 59.98 | |
LOC_Os04g44570-F-qrt | CGCCACCACTGGGTTTACT | 60.96 | 123 |
LOC_Os04g44570-R-qrt | CACGGGAAGCCGAGTATCT | 60.23 |
表1 实时荧光定量PCR的引物序列
Table 1 The primer sequences of real-time quantitative PCR
Primer name | Sequence (5′-3′) | Tm (°C) | Length (bp) |
---|---|---|---|
LOC_Os02g56630-F-qrt | GATGCTACGGAGACTGCACA | 60.02 | 182 |
LOC_Os02g56630-R-qrt | GAGCCAGAAGCACATGAACA | 59.99 | |
LOC_Os02g57720-F-qrt | CTGACGTCGTGGTCGTTCTA | 59.90 | 101 |
LOC_Os02g57720-R-qrt | GACTTGTTCACCCCCATCAC | 60.22 | |
LOC_Os04g43730-F-qrt | ACCAAAGTGGGTGTTTCTCG | 60.01 | 153 |
LOC_Os04g43730-R-qrt | TCGAGATCTGGCTTGTGTTG | 59.98 | |
LOC_Os04g44060-F-qrt | CGGTGTTCATGGTTCACTTG | 60.00 | 182 |
LOC_Os04g44060-R-qrt | CCTCAGGACGTACTGGTGGT | 60.03 | |
LOC_Os04g46490-F-qrt | GCCTCGTCCTCCACTACATC | 59.69 | 123 |
LOC_Os04g46490-R-qrt | CCGTGTACACCACCATGAAC | 59.73 | |
LOC_Os02g57580-F-qrt | ACCTCTTCAGATGGGGTGTG | 59.96 | 101 |
LOC_Os02g57580-R-qrt | CCAGTCAGTTTTGCAGACCA | 59.87 | |
LOC_Os04g48230-F-qrt | GGGCACTACAAGTCCGTGAT | 60.00 | 199 |
LOC_Os04g48230-R-qrt | CTTGGTAGCTTCCGATGAGC | 59.98 | |
LOC_Os04g44570-F-qrt | CGCCACCACTGGGTTTACT | 60.96 | 123 |
LOC_Os04g44570-R-qrt | CACGGGAAGCCGAGTATCT | 60.23 |
QTL | Chromosome | Physical distance (bp) | Position of support (cM) | Likelihood of odd (LOD) |
---|---|---|---|---|
qLpw2-8 | 2 | 34636269-35675126 | 148.48-152.93 | 3.27 |
qLpw3-8 | 3 | 30389734-30481217 | 130.27-130.66 | 2.93 |
qLpw4-8 | 4 | 24066261-30847136 | 103.17-132.23 | 5.15 |
qLpw11-8 | 11 | 2051418-2119803 | 8.79-9.09 | 3.06 |
qLpw12-8 | 12 | 25396321-25626163 | 108.87-109.85 | 3.34 |
表2 水稻分蘖期叶片水势的QTL分析
Table 2 QTL analysis of leaf water potential in rice at tillering stage
QTL | Chromosome | Physical distance (bp) | Position of support (cM) | Likelihood of odd (LOD) |
---|---|---|---|---|
qLpw2-8 | 2 | 34636269-35675126 | 148.48-152.93 | 3.27 |
qLpw3-8 | 3 | 30389734-30481217 | 130.27-130.66 | 2.93 |
qLpw4-8 | 4 | 24066261-30847136 | 103.17-132.23 | 5.15 |
qLpw11-8 | 11 | 2051418-2119803 | 8.79-9.09 | 3.06 |
qLpw12-8 | 12 | 25396321-25626163 | 108.87-109.85 | 3.34 |
Chromosome | Gene | Function | Regulation object | Reference |
---|---|---|---|---|
2 | LOC_Os02g56630 | Play an important regulatory role in the response of plants to abiotic stress | OsWAK24-OsWAK receptor-like protein kinase | |
4 | LOC_Os04g43730 | Play an important regulatory role in the response of plants to abiotic stress | OsWAK51-OsWAK receptor-like protein kinase | |
2 | LOC_Os02g57580 | Maybe involved in anthocyanin vacuole storage, participate in osmotic adjustment | Anthocyanin permease | Aza-González et al., 2013 |
2 | LOC_Os02g57720 | Mediate the transport of water across the membrane, regulate water | Aquaporin | |
4 | LOC_Os04g46490 | Mediate the transport of water across the membrane, regulate water | Aquaporin | |
4 | LOC_Os04g44570 | Mediate the transport of water across the membrane, regulate water | Aquaporin | |
4 | LOC_Os04g44060 | Mediate the transport of water across the membrane, regulate water | Aquaporin | |
4 | LOC_Os04g48230 | Maybe involved in the perception, conduc- tion of plant dehydration and regulation of antidehydration substance synthesis | Dehydration response related protein | Shinozaki and Yamaguchi-Shinozaki, 1997 |
表3 候选基因的功能注释
Table 3 The function annotation of candidate genes
Chromosome | Gene | Function | Regulation object | Reference |
---|---|---|---|---|
2 | LOC_Os02g56630 | Play an important regulatory role in the response of plants to abiotic stress | OsWAK24-OsWAK receptor-like protein kinase | |
4 | LOC_Os04g43730 | Play an important regulatory role in the response of plants to abiotic stress | OsWAK51-OsWAK receptor-like protein kinase | |
2 | LOC_Os02g57580 | Maybe involved in anthocyanin vacuole storage, participate in osmotic adjustment | Anthocyanin permease | Aza-González et al., 2013 |
2 | LOC_Os02g57720 | Mediate the transport of water across the membrane, regulate water | Aquaporin | |
4 | LOC_Os04g46490 | Mediate the transport of water across the membrane, regulate water | Aquaporin | |
4 | LOC_Os04g44570 | Mediate the transport of water across the membrane, regulate water | Aquaporin | |
4 | LOC_Os04g44060 | Mediate the transport of water across the membrane, regulate water | Aquaporin | |
4 | LOC_Os04g48230 | Maybe involved in the perception, conduc- tion of plant dehydration and regulation of antidehydration substance synthesis | Dehydration response related protein | Shinozaki and Yamaguchi-Shinozaki, 1997 |
图4 水稻叶片水势相关候选基因表达量差异 **表示华占与热研2号基因表达量在0.01水平上差异显著。
Figure 4 Differences in the expression level of candidate genes involved in leaf water potential in rice ** indicate significant differences in genes expression level between HZ and Nekken2 at 0.01 level.
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