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作物学报 ›› 2024, Vol. 50 ›› Issue (12): 2962-2970.doi: 10.3724/SP.J.1006.2024.42021

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

过表达水稻OsPIN2通过减弱地上部重力反应增大分蘖角度

吴佳俊(), 涂燃冉, 张秋丽, 邹沁雯, 孙志豪, 王宏, 何光华()   

  1. 西南大学水稻研究所 / 西南大学农业科学研究院 / 作物分子改良重庆市重点实验室, 重庆 400715
  • 收稿日期:2024-04-22 接受日期:2024-08-15 出版日期:2024-12-12 网络出版日期:2024-08-29
  • 通讯作者: *何光华, E-mail: heghswu@163.com
  • 作者简介:E-mail: wjj15310616335@163.com
  • 基金资助:
    重庆市自然科学基金创新群体(cstc2021jcyj-cxttX0004);重庆市现代农业产业技术体系水稻创新团队项目(CQMAITS202301)

Overexpression of OsPIN2 increases tiller angle by reducing shoot gravitropic response in rice

WU Jia-Jun(), TU Ran-Ran, ZHANG Qiu-Li, ZOU Qin-Wen, SUN Zhi-Hao, WANG Hong, HE Guang-Hua()   

  1. Rice Research Institute, Southwest University / Academy of Agricultural Sciences, Southwest University / Key Laboratory of Crop Molecular Improvement, Chongqing 400715, China
  • Received:2024-04-22 Accepted:2024-08-15 Published:2024-12-12 Published online:2024-08-29
  • Contact: *E-mail: heghswu@163.com
  • Supported by:
    National Natural Science Foundation of Chongqing, China(cstc2021jcyj-cxttX0004);Chongqing Modern Agricultural Industry Technology System Rice Innovation Team of Chongqing, China(CQMAITS202301)

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摘要：

地上部向重力性与分蘖角度形成紧密相关, 解析其调控机制有助于合理设计分蘖角度, 从而促进作物株型改良。在水稻生长素输出载体基因OsPIN2过表达株系中鉴定到2个分蘖角度显著增大的株系OE-OsPIN2-1/2。扫描电镜观察显示OE-OsPIN2-1/2分蘖基部的近地端与远地端近乎对称生长。向重力性检测发现OE-OsPIN2-1/2幼苗地上部重力反应降低, 且重力刺激后生长素标志基因OsIAA20和WUSCHEL相关同源盒基因WOX6/11不对称表达减弱, 表明过表达OsPIN2减弱重力作用下的生长素不对称分布。此外, OE-OsPIN2-1/2分蘖基部参与重力反应的正调控基因下调表达而负调控基因上调表达, 进一步表明过表达OsPIN2能够减弱地上部重力反应。本研究结果揭示了OsPIN2通过调控地上部向重力性从而控制水稻分蘖角度的机制, 为深入研究植物地上部重力反应提供理论依据。

关键词: 水稻, OsPIN2, 分蘖角度, 地上部向重力性

Abstract:

Shoot gravitropism is closely related to the formation of the tiller angle, and understanding its regulatory mechanism is crucial for rationally designing tiller angles to improve crop plant architecture. In the rice auxin efflux carrier gene OsPIN2 overexpression lines, two lines with significantly increased tiller angles, OE-OsPIN2-1 and OE-OsPIN2-2, were identified. Scanning electron microscopy revealed that the near-ground and far-ground parts of the tiller bases in OE-OsPIN2-1/2 grew nearly symmetrically. Shoot gravitropism assays indicated that the shoot gravitropic responses of OE-OsPIN2-1/2 seedlings were reduced, and the asymmetric expression of the auxin marker gene OsIAA20 and WUSCHEL RELATED HOMEOBOX6/11 (WOX6/11) was weakened upon gravistimulation. This suggests that overexpression of OsPIN2 attenuates the asymmetric distribution of auxin following gravistimulation. Furthermore, the expressions of positive regulators involved in the gravitropic response at the tiller base in OE-OsPIN2-1/2 lines were downregulated, while the expressions of negative regulators were upregulated. This further indicates that overexpression of OsPIN2 leads to a reduced shoot gravitropic response. This study elucidates the mechanism by which OsPIN2 controls rice tiller angle by regulating shoot gravitropism, providing a theoretical basis for in-depth studies of shoot gravitropic responses.

Key words: rice, OsPIN2, tiller angle, shoot gravitropism

表1

本研究中所用引物"

名称 Name 上游引物 Upstream primer (5′-3′) 下游引物 Downstream primer (5′-3′)
OsPIN2-OE GCAGGATCCCCGGGTACCATGATCACCGGACGCGAC AATGTTTGAACGGAGCTCCTATATCCCAAGAAGCACATAGTAG
OsPIN2-qPCR CAACACCTACTCCAGCCTC TGGACCAGTCAAGAACCTC
UBQ-qPCR GCTCCGTGGCGGTATCAT CGGCAGTTGACAGCCCTAG
OsIAA20-qPCR TGGCGGATATGTGAAGGTGAA TATGAGCCGAGGATGGACAAG
WOX6-qPCR TCCAATAGACTTGCGAGCCAT GCATTAGGATTCCATAGTCGTT
WOX11-qPCR CGGTGTTCATCAACGGAGTG TCTGGAGAGAATGGAGGAGGAT
CRCT-qPCR TTCTGGGTGCCTCAACTCA AACGCTGTCTCAAAGTCCAATC
FucT-qPCR GGAGTCTGCTGTGCTTGCTA ACTGGTATAATGCCTGTCGTTGTG
AGPL1-qPCR TTGATTCCACATGGCAGAGAAC GTTGCTGCTGCTACTTCACT
OsHOX1-qPCR AGCACAACACCCTCAATC GTTCTGGAACCACACCTC
OsHOX28-qPCR CATTGACCACCCTCACAA CATTGACCACCCTCACAA
LPA1-qPCR GCGTATGTATGTAAAGCAAG GAAACGACCTACGAAACTAC
LA1-qPCR GAGATGAACGGCAACAAG TTCCAGCACCAAGTAGTC
LA2-qPCR GAACCAGCAGCCTGTAAGA AGCCATCCTCTCCTTCATTG
LA3-qPCR GGCGTCGCGCCAGCCTCA CTTCGGCGGAGTATCACG
TAC1-qPCR AGATGGCTCTAAAGGTGTTCAA TCTTCCATGGCCTTGTTCTC
TAC4-qPCR AAGGTCGCAAACAAGCAG AACTGCCAGGAGCAGAGAG
HSFA2D-qPCR CAGCAGGCACTTGGCACC TTCTTGTCACGCTTTAGCCTGT
OsARF12-qPCR GTTGGGAGGTCGTTGGACATAA AAGCACATCATTCTCCCTGTCG
OsARF17-qPCR TTTACAAATCGGGAACCTATGG TTTATGCAGGAGACGCTATTCA
OsARF25-qPCR TGACATCTCCAGATTCAGCAGC CGTCTCCACCACGAACCAA

图1

野生型与OE-OsPIN2-1/2的表型分析 A: 野生型与OE-OsPIN2-1/2分蘖期植株对比, 标尺为20 cm; B: 野生型与OE-OsPIN2-1/2分蘖基部OsPIN2表达量分析(n = 3); C: 野生型与OE-OsPIN2-1/2分蘖角度对比, 标尺为4 cm; D: 野生型与OE-OsPIN2-1/2分蘖角度测量(n = 10)。平均值±标准差, 双尾Student’s t检验, **: P < 0.01。"

图2

野生型与OE-OsPIN2-1/2的分蘖基部分析 A~C: 野生型与OE-OsPIN2-1/2抽穗期分蘖基部对比, 标尺为1 cm; D~F: 野生型与OE-OsPIN2-1/2分蘖基部近地端细胞的扫描电镜观察, 标尺为50 μm; G~I: 野生型与OE-OsPIN2-1/2分蘖基部远地端地端细胞的扫描电镜观察, 标尺为50 μm; J~K:野生型与OE-OsPIN2-1/2分蘖基部近地端和远地端细胞长度(J)和宽度(K)的测量(n = 20); L: 野生型与OE-OsPIN2-1/2分蘖基部近地端和远地端单位面积细胞数目统计(n = 3)。NG表示近地端(near-ground part), FG表示远地端(far-ground part)。平均值±标准差, 双尾Student’s t检验, **: P < 0.01。"

图3

野生型与OE-OsPIN2-1/2幼苗的地上部向重力性检测 A: 野生型与OE-OsPIN2-1/2幼苗重力刺激0 h、12 h、24 h、36 h和48 h后植株对比; B: 野生型与OE-OsPIN2-1/2幼苗重力刺激0 h、12 h、24 h、36 h和48 h后植株茎秆弯曲度测量(n = 5)。平均值±标准差。多重比较采用Duncan’s检验, 不同字母(B中小写字母a和b)代表在0.05水平差异显著。"

图4

重力刺激前后OsIAA20、WOX6/11以及分蘖基部分蘖角度相关基因表达量分析 A: 用于表示重力刺激后幼苗茎基部下端(lower site, LS)和上端(upper site, US)的示意图; g代表重力, 箭头代表重力方向; B: 重力刺激0 h和重力刺激6 h, 野生型和OE-OsPIN2-1/2幼苗茎基部下端和上端生长素标志基因OsIAA20和WUSCHEL相关同源盒基因WOX6/11的转录水平分析; C: 野生型和OE-OsPIN2-1/2分蘖期分蘖基部分蘖角度相关基因表达量分析; D: 地上部重力反应调控水稻分蘖角度网络图[14,30]; 红色向上箭头表示该基因上调表达,红色向下箭头表达该基因下调表达。平均值±标准差(n = 3), 双尾Student’s t检验, **: P < 0.01。"

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