植物学报 ›› 2022, Vol. 57 ›› Issue (5): 588-595.DOI: 10.11983/CBB22114
魏和平1,†, 芦涛2,†, 贾绮玮2, 邓飞3, 朱浩3, 岂泽华1, 王玉玺1, 叶涵斐2, 殷文晶2, 方媛2, 穆丹1,*(), 饶玉春2,*(
)
收稿日期:
2022-05-31
接受日期:
2022-07-25
出版日期:
2022-09-01
发布日期:
2022-09-09
通讯作者:
穆丹,饶玉春
作者简介:
ryc@zjnu.cn基金资助:
Wei Heping1,†, Lu Tao2,†, Jia Qiwei2, Deng Fei3, Zhu Hao3, Qi Zehua1, Wang Yuxi1, Ye Hanfei2, Yin Wenjing2, Fang Yuan2, Mu Dan1,*(), Rao Yuchun2,*(
)
Received:
2022-05-31
Accepted:
2022-07-25
Online:
2022-09-01
Published:
2022-09-09
Contact:
Mu Dan,Rao Yuchun
About author:
ryc@zjnu.cn摘要:
水稻(Oryza sativa)抽穗期是决定产量和品质的重要性状, 在育种、制种及引种驯化过程中发挥重要作用。将热研2号(O. sativa subsp. japonica cv. ‘Nekken2’)和华占(O. sativa subsp. indica cv. ‘HZ’)杂交获得F1代, 经连续多代自交得到120个重组自交系(RILs)群体。在常规水肥管理条件下, 对120个RILs株系的抽穗时间进行统计分析。利用已构建好的高密度遗传图谱, 对水稻抽穗期相关性状进行QTL定位分析, 结果共检测到11个QTLs, 分别位于第1、3、4、5、6、8和12号染色体上, 其中1个LOD值高达5.75。通过分析QTLs区间内的候选基因, 筛选出可能影响两亲本抽穗期的相关基因, 并利用实时定量PCR进行基因表达量分析, 发现LOC_Os03g03070、LOC_Os03g50310、LOC_Os03g55389、LOC_Os04g55510、LOC_Os08g07740和LOC_Os08g01670共6个基因在双亲间的表达量差异显著, 其中LOC_Os03g50310在Nekken2中的表达量比HZ高3.6倍。对双亲间候选基因LOC_Os03g50310进行测序分析, 发现该基因在5′UTR、CDS区及3′UTR存在4处差异, 其中CDS区的单核苷酸多态性(SNP)差异引发单个氨基酸改变。研究通过挖掘水稻抽穗期QTL位点为进一步克隆水稻抽穗期相关基因和品种选育提供了新线索。
魏和平, 芦涛, 贾绮玮, 邓飞, 朱浩, 岂泽华, 王玉玺, 叶涵斐, 殷文晶, 方媛, 穆丹, 饶玉春. 水稻抽穗期QTL定位及候选基因分析. 植物学报, 2022, 57(5): 588-595.
Wei Heping, Lu Tao, Jia Qiwei, Deng Fei, Zhu Hao, Qi Zehua, Wang Yuxi, Ye Hanfei, Yin Wenjing, Fang Yuan, Mu Dan, Rao Yuchun. QTL Mapping of Candidate Genes for Heading Date in Rice. Chinese Bulletin of Botany, 2022, 57(5): 588-595.
Primer name | Forward primer (5′-3′) | Reverse primer (5′-3′) |
---|---|---|
OsActin | TGGCATCTCAGCACATTCC | TGCACAATGGATGGGTCAGA |
LOC_Os03g50310 | AGCAGCAGATCGAGCAGAAG | CTTCTCGTCGTCCAAGAAGC |
LOC_Os03g55389 | TGTCAAGCCCCACAATGTTA | AAGGCTCCACATGTCCAAAG |
LOC_Os03g57940 | AGCACCGAAGCAGTAAAGGA | AAGCAACCATATGGGCAGAC |
LOC_Os03g03070 | AAAAGCTGACGCTGATGGTT | TCCTTCCCCTGATGCTAATG |
LOC_Os04g55510 | GTGGCCTAAGGATTGGTGAA | TGAGAGCAGCCACCATGTAG |
LOC_Os08g07740 | GGAAGAGCTATGGGCACTTG | CGACTCCTTGGAGATCTTGG |
LOC_Os08g01420 | GACAGGTCTAGCGAGCAACC | GGCACAGTTCGTTTCGGTAT |
LOC_Os08g01670 | GATGGGGCAGCAGGACTAC | GATGCAGGAGAGGAGGTGAG |
表1 qRT-PCR引物序列
Table 1 qRT-PCR primer sequences
Primer name | Forward primer (5′-3′) | Reverse primer (5′-3′) |
---|---|---|
OsActin | TGGCATCTCAGCACATTCC | TGCACAATGGATGGGTCAGA |
LOC_Os03g50310 | AGCAGCAGATCGAGCAGAAG | CTTCTCGTCGTCCAAGAAGC |
LOC_Os03g55389 | TGTCAAGCCCCACAATGTTA | AAGGCTCCACATGTCCAAAG |
LOC_Os03g57940 | AGCACCGAAGCAGTAAAGGA | AAGCAACCATATGGGCAGAC |
LOC_Os03g03070 | AAAAGCTGACGCTGATGGTT | TCCTTCCCCTGATGCTAATG |
LOC_Os04g55510 | GTGGCCTAAGGATTGGTGAA | TGAGAGCAGCCACCATGTAG |
LOC_Os08g07740 | GGAAGAGCTATGGGCACTTG | CGACTCCTTGGAGATCTTGG |
LOC_Os08g01420 | GACAGGTCTAGCGAGCAACC | GGCACAGTTCGTTTCGGTAT |
LOC_Os08g01670 | GATGGGGCAGCAGGACTAC | GATGCAGGAGAGGAGGTGAG |
QTL | Chromosome | Physical distance (bp) | Position of support (cM) | Likelihood of odd (LOD) |
---|---|---|---|---|
qhd-1 | 1 | 25257953-25481977 | 108.27-109.23 | 3.75 |
qhd-3 | 3 | 13767411-14882947 | 54.53-63.79 | 3.82 |
qhd-4 | 3 | 15628988-15779451 | 66.72-67.64 | 3.09 |
qhd-5 | 3 | 17197291-17357469 | 73.72-74.40 | 3.33 |
qhd-6 | 3 | 28536151-33494350 | 122.32-143.58 | 4.87 |
qhd-7 | 4 | 2572479-3906500 | 11.02-16.74 | 3.17 |
qhd-8 | 4 | 31066905-34944379 | 133.17-149.79 | 5.75 |
qhd-9 | 5 | 26262798-26850392 | 112.58-115.10 | 3.14 |
qhd-10 | 6 | 4227455-4659759 | 18.12-19.97 | 3.97 |
qhd-11 | 8 | 3480879-4936569 | 18.12-19.97 | 5.06 |
qhd-12 | 12 | 26219257-27272729 | 112.39-116.97 | 3.98 |
表2 水稻抽穗期QTL分析
Table 2 Analysis of QTL for heading date in rice
QTL | Chromosome | Physical distance (bp) | Position of support (cM) | Likelihood of odd (LOD) |
---|---|---|---|---|
qhd-1 | 1 | 25257953-25481977 | 108.27-109.23 | 3.75 |
qhd-3 | 3 | 13767411-14882947 | 54.53-63.79 | 3.82 |
qhd-4 | 3 | 15628988-15779451 | 66.72-67.64 | 3.09 |
qhd-5 | 3 | 17197291-17357469 | 73.72-74.40 | 3.33 |
qhd-6 | 3 | 28536151-33494350 | 122.32-143.58 | 4.87 |
qhd-7 | 4 | 2572479-3906500 | 11.02-16.74 | 3.17 |
qhd-8 | 4 | 31066905-34944379 | 133.17-149.79 | 5.75 |
qhd-9 | 5 | 26262798-26850392 | 112.58-115.10 | 3.14 |
qhd-10 | 6 | 4227455-4659759 | 18.12-19.97 | 3.97 |
qhd-11 | 8 | 3480879-4936569 | 18.12-19.97 | 5.06 |
qhd-12 | 12 | 26219257-27272729 | 112.39-116.97 | 3.98 |
Chromosome | Gene name | Function |
---|---|---|
3 | LOC_Os03g50310 | Regulation of rice blast resistance through salicylic acid and ethylene signaling pathways, mediating heading stage |
3 | LOC_Os03g55389 | Delaying flowering time by inhibiting the expression of FT-like genes |
3 | LOC_Os03g57940 | Negative regulation of gibberellin signaling by phosphorylating the DELLA protein SLR1 |
3 | LOC_Os03g03070 | Involved in the regulation of adventitious root formation in rice by regulating auxin transport |
4 | LOC_Os04g55510 | Mediates ubiquitination to degrade Hd1 through a 26S proteasome-mediated pathway |
8 | LOC_Os08g07740 | Controlling the yield, plant height and heading date of rice |
8 | LOC_Os08g01420 | Induction of rice heading by inhibiting Ghd7 |
8 | LOC_Os08g01670 | Regulation of pectin methyl esterification levels |
表3 候选基因功能
Table 3 The function of candidate genes
Chromosome | Gene name | Function |
---|---|---|
3 | LOC_Os03g50310 | Regulation of rice blast resistance through salicylic acid and ethylene signaling pathways, mediating heading stage |
3 | LOC_Os03g55389 | Delaying flowering time by inhibiting the expression of FT-like genes |
3 | LOC_Os03g57940 | Negative regulation of gibberellin signaling by phosphorylating the DELLA protein SLR1 |
3 | LOC_Os03g03070 | Involved in the regulation of adventitious root formation in rice by regulating auxin transport |
4 | LOC_Os04g55510 | Mediates ubiquitination to degrade Hd1 through a 26S proteasome-mediated pathway |
8 | LOC_Os08g07740 | Controlling the yield, plant height and heading date of rice |
8 | LOC_Os08g01420 | Induction of rice heading by inhibiting Ghd7 |
8 | LOC_Os08g01670 | Regulation of pectin methyl esterification levels |
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