不同磷胁迫处理转OsPHR2小麦的转录组学分析

doi:10.3724/SP.J.1006.2024.31020

作物学报 ›› 2024, Vol. 50 ›› Issue (2): 340-353.doi: 10.3724/SP.J.1006.2024.31020

• 作物遗传育种·种质资源·分子遗传学 • 上一篇下一篇

不同磷胁迫处理转OsPHR2小麦的转录组学分析

李艳(), 方宇辉, 王永霞, 彭超军, 华夏, 齐学礼, 胡琳, 许为钢^*()

河南省作物分子育种研究院 / 河南省麦类种质资源创新与改良重点实验室 / 神农种业实验室, 河南郑州 450002

收稿日期:2023-03-20 接受日期:2023-06-29 出版日期:2024-02-12 网络出版日期:2023-07-17
通讯作者: *许为钢, E-mail: xuwg1958@163.com
作者简介:E-mail: liyanly7812@163.com
基金资助:
国家自然科学基金项目(31701510);神农种业实验室“一流课题”项目(SN01-2022-01)

Transcriptomics profile of transgenic OsPHR2 wheat under different phosphorus stress

LI Yan(), FANG Yu-Hui, WANG Yong-Xia, PENG Chao-Jun, HUA Xia, QI Xue-Li, HU Lin, XU Wei-Gang^*()

Henan Academy of Crops Molecular Breeding / Key Laboratory for Innovation and Improvement of Triticeae Germplasm Resources of Henan Province / Shennong Laboratory, Zhengzhou 450002, Henan, China

Received:2023-03-20 Accepted:2023-06-29 Published:2024-02-12 Published online:2023-07-17
Contact: *E-mail: xuwg1958@163.com
Supported by:
National Natural Science Foundation of China(31701510);‘First-class Project’ of Shennong Laboratory(SN01-2022-01)

摘要/Abstract

摘要：

PHR基因是磷信号调控体系的核心转录因子, 负责启动下游部分应对磷饥饿的适应性反应。本课题前期获得了磷高效转OsPHR2小麦纯系, 但OsPHR2提高小麦磷吸收利用效率的分子机制尚不清楚。为揭示该机制, 本研究以前期获得的磷高效转OsPHR2小麦纯系为研究材料, 采用水培试验, 小麦长至四叶一心时进行低磷胁迫处理, 分别在低磷胁迫0、6、24和72 h, 利用RNA-Seq进行转录组测定, 分析转基因小麦与对照之间根部和叶片的差异表达基因(differentially expression gene, DEG), 并分别对根部和叶片DEG进行GO和KEGG功能富集分析。结果显示: 低磷胁迫处理0、6、24和72 h转基因系与对照根部有22个共同的DEG, 叶片有9个共同的DEG。转基因小麦与对照根部DEG数量在低磷胁迫处理0 h最多, 其次为6 h。GO和KEGG富集分析显示, 低磷胁迫0 h和6 h, 根部DEG主要富集在糖代谢、苯丙素生物合成等生物学过程, 以及养分贮存器活性、ATP酶活性等分子功能。转基因小麦与对照叶片DEG数量在低磷胁迫72 h最多, 主要富集在糖代谢、有机酸生物合成等生物学过程, 以及与糖基转移酶活性、纤维素合酶活性等有关的分子功能。与对照相比, 转基因系OsT5-28根部血红素过氧化物酶、谷胱甘肽S-转移酶等防御系统关键酶基因, 以及叶片磷酸丙糖转运体家族基因在低磷胁迫前后均上调表达。转OsPHR2小麦与对照对低磷胁迫的响应程度具有一定的差异性, 低磷胁迫下转基因小麦较对照具有较强的磷素吸收利用能力, 主要是OsPHR2调控了小麦中相关基因表达。

关键词: 低磷胁迫, 转基因小麦, 转录组, 磷素吸收利用效率, 差异表达基因

Abstract:

The PHR gene is the core transcription factor in the phosphorus signaling regulatory system, responsible for initiating the adaptive response of downstream parts to phosphorus starvation. At the early stage, the transgenic OsPHR2 wheat pure lines with high phosphorus efficiency were obtained, but the molecular mechanism of OsPHR2 improving the phosphorus absorption and utilization efficiency of wheat is still unclear. In order to reveal the molecular mechanism of OsPHR2 improving the phosphorus uptake and utilization efficiency in wheat, transgenic OsPHR2 wheat pure line with high phosphorus efficiency earlier as the experimental material in this study. Transgenic OsPHR2 wheat and the control were treated with low phosphorus stress when they grew to 4 leaves and 1 heart in hydroponics experiment. Transgenic OsPHR2 wheat and control under low phosphorus stress for 0, 6, 24, and 72 h were used for transcriptomes analysis by RNA-seq. The differentially expression genes (DEGs) in roots and leaves of transgenic wheat and control were analyzed. There were 22 common DEGs in the roots of transgenic wheat and control under low phosphorus stress for 0, 6, 24, and 72 h, and there were nine common DEGs in the leaves under four treatments. The functional and pathway enrichments of differentially expressed genes in roots and leaves were also performed by GO and KEGG analysis. The result showed that the number of DEGs in the root of transgenic wheat and control was the highest under low phosphorus stress for 0 h, followed by 6 h. GO and KEGG enrichment analysis suggested that DEGs were mainly clustered into biological processes such as glucose metabolism, phenylpropanoid biosynthesis, and molecular functions such as nutrient storage activity, ATPase activity, etc. The number of DEGs in the leaves of transgenic wheat and the control was the highest under low phosphorus stress for 72 h. DEGs were mainly clustered into biological processes such as glucose metabolism, organic acid biosynthesis, as well as molecular functions related to glycosyltransferase activity and cellulose synthase activity. Compared with the control, the key enzyme genes of the defense system such as heme peroxidase and glutathione S-transferase in the root of the transgenic line OsT5-28, as well as the trisphosphate transporter family genes in the leaf were up-regulated before and after low phosphorus stresses. The response of transgenic OsPHR2 wheat to low phosphorus stress was different from that of the control. Transgenic wheat had stronger phosphorus absorption and utilization ability than the control under low phosphorus stress, mainly because OsPHR2 regulated the relative expression level of related genes in wheat.

Key words: low phosphorus stress, transgenic wheat, transcriptomics, phosphorus absorption and utilization efficiency, differentially expressed genes

李艳, 方宇辉, 王永霞, 彭超军, 华夏, 齐学礼, 胡琳, 许为钢. 不同磷胁迫处理转OsPHR2小麦的转录组学分析[J]. 作物学报, 2024, 50(2): 340-353.

LI Yan, FANG Yu-Hui, WANG Yong-Xia, PENG Chao-Jun, HUA Xia, QI Xue-Li, HU Lin, XU Wei-Gang. Transcriptomics profile of transgenic OsPHR2 wheat under different phosphorus stress[J]. Acta Agronomica Sinica, 2024, 50(2): 340-353.

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基因名称 Gene ID	上游引物序列 Forward sequence (5′-3′)	下游引物序列 Reverse sequence (5′-3′)
TraesCS7A02G173000 (叶Leaf)	CAACAAGTTCTTGCCGACCA	CTATCCCTTGCATGCACGTC
TraesCS6A02G058200 (叶Leaf)	TGTTTCTCTGTCAGGGCCAA	TTGTTTCTTGCTCGTCGGTG
TraesCS6A02G059600 (叶Leaf)	GGAGTACCCCAACCTGTTCA	CCTTTCTCTGTCATGTGGCG
TraesCS6A02G052400 (叶Leaf)	GTCCGGTTTAGTTGGGGAGA	TTCACCACCATGTACTCCCC
TraesCS6B02G465800 (叶Leaf)	CACGGTGCCTAGAGTTGTTG	TTCAACGTTTCCAGTGCCTG
TraesCS6A02G059000 (根Root)	GACAACATCCAGGGCATCAC	GGATGACGTTCTCCAGGAAG
TraesCS6A02G055200 (根Root)	CAACATCCAGGGCATCAC	GGATGACGTTCTCCAGGAAG
TraesCS6A02G035000 (根Root)	AGAAGGGCGTGGAGACCTAC	GTGATGGTGGGCTTCTTGTT
novel.11254 (根Root)	CAGCAGTTCAAGATCCACGA	CCAATCCAATACCTCCCTGA
TraesCS6A02G035200 (根Root)	TGCCAAGGACAACAAGAAGA	GGAGTTGATGTTGGGGATCA
Actin	ATGTTGTTCTCAGTGGAGGTTC	CTGTATTTCCTTTCAGGTGGTGC

基因ID Gene	名称 Description	R_0h_T vs R_0h_C up	R_0h_T vs R_0h_C Down	R_6h_T vs R_6h_C up	R_6h_T vs R_6h_C Down
dosa00592	α-亚麻酸代谢Alpha-Linolenic acid metabolism	1	6	0	0
dosa00480	谷胱甘肽代谢Glutathione metabolism	0	0	8	2
dosa00564	甘油磷脂代谢Glycerophospholipid metabolism	2	11	0	0
dosa00940	苯丙类生物合成Phenylpropanoid biosynthesis	12	22	9	1
dosa04075	植物激素信号转导Plant hormone signal transduction	3	21	0	0

基因ID Gene ID	名称 Description	L_0 h_T vs L_0 h_C Down	L_72 h_T vs L_72 h_C up
dosa00062	脂肪酸伸长率Fatty acid elongation	0	5
dosa00941	类黄酮生物合成Flavonoid biosynthesis	0	5
dosa00360	苯丙氨酸代谢Phenylalanine metabolism	0	5
dosa00940	苯丙类生物合成Phenylpropanoid biosynthesis	0	11
dosa00196	光合作用-天线蛋白Photosynthesis-antenna proteins	2	0

不同磷胁迫处理转OsPHR2小麦的转录组学分析

Transcriptomics profile of transgenic OsPHR2 wheat under different phosphorus stress

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