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Acta Agronomica Sinica ›› 2024, Vol. 50 ›› Issue (2): 340-353.doi: 10.3724/SP.J.1006.2024.31020

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

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*()   

  1. 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 Online:2024-02-12 Published: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)

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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

Fig. 1

Principal component analysis of RNA-Seq data from wheat under different phosphorus treatments"

Table 1

Information of qRT-PCR primers"

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

Fig. 2

DEGs in roots between transgenic wheat and control in different comparative combinations A: The numbers of DEGs between transgenic wheat and control; B: Venn diagram of DEGs between transgenic wheat and control."

Fig. 3

DEGs in roots between 6 hours and 0 hour under low phosphorus in control"

Fig. 4

DEGs in leaves between transgenic wheat and control in different comparative combinations A: The numbers of DEGs between transgenic wheat and control; B: Venn diagram of DEGs between transgenic wheat and control."

Fig. 5

DEGs in leaves between 72 hours and 0 hour under low phosphorus in control"

Fig. 6

qRT-PCR verification of selected DEGs in root and leaf under different phosphorus treatments 1: TraesCS7A02G173000 (Leaf); 2: TraesCS6A02G058200 (Leaf); 3: TraesCS6A02G059600 (Leaf); 4: TraesCS6A02G052400 (Leaf); 5: TraesCS6B02G465800 (Leaf); 6: TraesCS6A02G059000 (Root); 7: TraesCS6A02G055200 (Root); 8: TraesCS6A02G035000 (Root); 9: novel.11254 (Root); 10: TraesCS6A02G035200 (Root)."

Fig. 7

GO enrichment of root DEGs between transgenic wheat and control in different comparative combinations"

Fig. 8

GO enrichment of root DEGs between 6 hours and 0 hour under low phosphorus in control"

Table 2

KEGG enrichment of root DEGs in different comparative combinations"

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

Fig. 9

KEGG enrichment of root DEGs between 6 hours and 0 hour under low phosphorus in control"

Fig. 10

GO enrichment of leaf DEGs between transgenic wheat and control in different comparative combinations"

Fig. 11

GO enrichment of leaf DEGs between 72 hours and 0 hour under low phosphorus in control"

Table 3

KEGG enrichment of leaf DEGs in different comparative combinations"

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

Fig. 12

KEGG enrichment of leaf DEGs between 72 hours and 0 hour under low phosphorus in control"

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