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Acta Agronomica Sinica ›› 2023, Vol. 49 ›› Issue (7): 1808-1817.doi: 10.3724/SP.J.1006.2023.21050

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

Transcriptome analysis of exogenous 6-BA in regulating young spike development of wheat after low temperature at booting stage

LI Ling-Yu(), ZHOU Qi-Rui, LI Yang, ZHANG An-Min, WANG Bei-Bei, MA Shang-Yu, FAN Yong-Hui, HUANG Zheng-Lai*(), ZHANG Wen-Jing*()   

  1. College of Agronomy, Anhui Agricultural University, Hefei 230036, Anhui, China
  • Received:2022-07-17 Accepted:2022-11-25 Online:2023-07-12 Published:2022-12-01
  • Contact: *E-mail: zhangwenjing79@126.com; E-mail: xdnyyjs@163.com E-mail:171650776@qq.com;xdnyyjs@163.com;zhangwenjing79@126.com
  • Supported by:
    The National Natural Science Foundation of China(31801285);The Anhui Key Research and Development Plan(202204c06020040);The Major Science and Technology Project of Anhui Province(202003a06020014)

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Abstract:

In recent years, due to the frequent occurrence of extreme weather, low temperatures (LT) became one of the main disasters restricting wheat production. LT has negative effects on wheat growth and yield formation, especially at booting stage. Exogenous spraying of 6-benzylamino adenine (6-BA) after LT at booting stage can alleviate the damage caused by LT in wheat, but the related molecular regulation mechanism is still unclear. In this study, transcriptome sequencing technology was used to analyze the molecular mechanism of 6-BA improving cold tolerance in wheat. The LT sensitive variety Wanmai 52 and insensitive variety Yannong 19 were selected as the experimental materials. The experiment was carried out in combination of potted and field planting. Two wheat cultivars were planted in plastic pots at a planting density of ten plants per pot. At booting stage, the pots were moved into an artificial climate chamber for low temperature treatment. At the end of the treatment, 20 mg L-1 6-BA solution was sprayed, and an equal volume of distilled water was sprayed as the control. The morphology of young spike, and the content of soluble sugar and starch in young spikes were determined. Some candidate differentially expressed genes (DEGs) were screened and their relative expression patterns were analyzed by qRT-PCR, and the results were verified by qRT-PCR. After 10 days, compared with the control, the morphological development of young spike was better, fuller and longer. The contents of soluble sugar and starch in young spikes increased after 6-BA treatment. The results showed that 22,770 DEGs were identified in Wanmai 52 and 9866 in Yannong 19, respectively, and 661 genes were up-regulated in the two cultivars. ARF5, AGPL1, 1-SST, SWEET15, and other genes were screened out, which were related to the regulation of plant hormone level, starch synthesis, and sugar metabolism. GO and KEGG enrichment analysis were performed on the selected differential genes. GO annotation revealed that the functions of the differential genes of the two varieties were mainly concentrated in cell structure stability, metabolism, and catalytic activity. KEGG enrichment analysis demonstrated that signal transduction, regulation of endogenous hormone levels, carbon metabolism, the changes of membrane structure and function had significant changes. In conclusion, 6-BA could alleviate cold damage by regulating the metabolism of antioxidant substances, hormone signal transduction, carbohydrate metabolism, osmotic adjustment, and other ways in wheat. The results provide a theoretical basis for exploring the cultivation measures to reduce the damage of low temperature on wheat in spring.

Key words: wheat, 6-BA, low temperatures, young spikes, transcriptome

Table 1

Primers for qRT-PCR used in this study"

基因名称
Gene name		 正向引物
Forward sequence (5′-3′)		 反向引物
Reverse sequence (5′-3′)		 基因ID
Gene ID
GAPDH CCACTAACTGCCTTGCTCCT CCAGTGCTGCTTGGAATGA
ARF5 TCGACAGAGTATGGGTTGGAA CGGAAACATCGGCTGAAG TraesCS6A02G111900
AGPL1 ACATCACGCAGAAACCTACCA AACGGATTCCAACGACACTG TraesCS5D02G484500
SWEET15 TATCAGTTGGTTTGTGTGTTCGA GTCGGGAGTGGTGAAAGGA TraesCS7B02G050500

Fig. 1

Phenotypic changes of young spike after exogenous 6-BA treatment Wanmai 52 CK is Wanmai 52 sprayed with water after low temperature treatment; Wanmai 52 6-BA is Wanmai 52 sprayed with 6-BA solution after low temperature treatment; Yannong 19 CK is Yannong 19 sprayed with water after low temperature treatment; Yannong 19 CK is Yannong 19 sprayed with 6-BA solution after low temperature treatment."

Fig. 2

Soluble sugar and starch content in young spikes"

Fig. 3

Venn plots of differently expressed genes (DEGs)"

Table 2

GO annotation of DEGs after spraying 6-BA under low temperature stress"

GO号
GO ID		 GO功能
GO function		 皖麦52基因数目
及其所占百分比
Gene number and ratio in Wanmai 52		 烟农19基因数目
及其所占百分比
Gene number and ratio in Yannong 19		 GO类别
GO category
GO:0016021 膜的有机成分Integral component of membrane 3990 (23.22%) 1367 (18.73%) C
GO:0005524 ATP结合ATP binding 2195 (12.77%) 764 (10.47%) M
GO:0005515 蛋白质结合Protein binding 2092 (12.17%) 920 (12.61%) M
GO:0016020 细胞膜Membrane 1609 (9.36%) 566 (7.76%) C
GO:0055114 氧化还原过程Oxidation reduction process 1604 (9.33%) 708 (9.70%) B
GO:0005634 细胞核Nucleus 1576 (9.17%) 1076 (14.74%) C
GO:0004672 蛋白激酶活性Protein kinase activity 1503 (8.75%) 428 (5.86%) M
GO:0006468 蛋白磷酸化Protein phosphorylation 1500 (8.73%) 427 (5.85%) B
GO:0003677 DNA结合DNA binding 1410 (8.20%) 1034 (14.17%) M
GO:0006355 调控转录, DNA模板化
Regulation of transcription, DNA templating		 1083 (6.30%) 410 (5.62%) B
GO:0003824 催化活性Catalytic activity 1005 (5.85%) 372 (5.10%) M
GO:0046872 金属离子结合Metal ion binding 953 (5.55%) 432 (5.92%) M
GO:0016491 氧化还原酶活性Oxidoreductase activity 901 (5.24%) 385 (5.28%) M
GO:0055085 跨膜转运Transmembrane transport 799 (4.65%) 266 (3.64%) B
GO:0003700 DNA结合转录因子活性
DNA-binding transcription factor activity		 648 (3.77%) 232 (3.18%) M
GO:0020037 血红素结合Heme binding 617 (3.59%) 301 (4.12%) M
GO:0005975 碳水化合物代谢过程Carbohydrate metabolic process 535 (3.11%) 256 (3.51%) B

Fig. 4

Scatter diagram of KEGG pathway enrichment"

Fig. 5

Analysis of gene expression pattern in IAA signaling pathway The color lump represents the corresponding FPKM value, and the higher the value, the redder the color. The samples from left to right are Wanmai 52 CK, Wanmai 52 6-BA, Yannong 19 CK, and Yannong 19 6-BA."

Fig. 6

Analysis of the expression pattern of carbohydrate accumulation related genes The contents of the colors are the same as those given in Fig. 5."

Fig. 7

Verification of the differentially expressed genes by qRT-PCR"

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