Scientia Agricultura Sinica ›› 2023, Vol. 56 ›› Issue (6): 1035-1044.doi: 10.3864/j.issn.0578-1752.2023.06.002

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

Investigation of Low Nitrogen Tolerance of ZmCCT10 in Maize

LI YiPu1,2(), TONG LiXiu3(), LIN YaNan1,2, SU ZhiJun1,2, BAO HaiZhu1,2, WANG FuGui2,4, LIU Jian2,4, QU JiaWei1,2, HU ShuPing2,4, SUN JiYing1,2, WANG ZhiGang1,2, YU XiaoFang1,2(), XU MingLiang3(), GAO JuLin1,2()   

  1. 1 Agricultural College, Inner Mongolia Agricultural University, Hohhot 010018
    2 Region Research Center for Conservation and Utilization of Crop Germplasm Resources in Cold and Arid Areas, Hohhot 010018
    3 College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193
    4 Vocational and Technical College of Inner Mongolia Agricultural University, Baotou 014109, Inner Mongolia
  • Received:2022-12-09 Accepted:2022-12-27 Online:2023-03-16 Published:2023-03-23

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Abstract:

【Objective】 The lack of soil nitrogen impacts the yield and quality of maize, which is a major problem of maize production in China. ZmCCT10 encodes the transcription factor, which is pleiotropic. ZmCCT10 is a very important co-factor regulating the growth, development and responding to abiotic stress of maize. The molecular mechanism of maize tolerance to low nitrogen is the basis for breeding maize varieties with low nitrogen tolerance and high nitrogen efficiency. 【Method】 In this study, we compared the traits those relate to low-nitrogen tolerance, expression pattern of ZmCCT10 and transcriptome results of ZmCCT10 near-isogenic lines under low-nitrogen stress and complete nutrient conditions. To analysis the characteristics of ZmCCT10 in response to low-nitrogen stress and the molecular mechanisms involved in low-nitrogen tolerance were explored. 【Result】 This study indicated that different alleles of ZmCCT10 showed significant differences in root length traits, biomass and nitrogen physiological traits under low nitrogen stress. The Y331-ΔTE haplotype without transposon insertion of ZmCCT10 had significantly longer total root length, main radicle length and lateral root length than Y331 after low nitrogen stress. What is more, root dry weight, shoot dry weight, nitrogen accumulation and nitrate reductase activity were also significantly higher than Y331. The expression levels of ZmCCT10 in roots and leaves were significantly higher than those in the control treatment. The expression level of ZmCCT10 in roots reached a peak at 3 hours after stress treatment. In leaves, the expression of ZmCCT10 continued to increase and peaked 6 hours after stress treatment. Root samples were collected under 0.04 mmol·L-1 low nitrogen stress after 3h for transcriptome sequencing. The correlation coefficients between biological replicates are more than 0.9. GO enrichment analysis showed that the expression levels of amine synthesis process and cellular nitrogen compound catabolic process were significantly different in near-isogenic lines after low nitrogen stress. Combined with the amount and expression pattern of differential genes, ZmCCT10-regulated candidate genes involved in low-nitrogen tolerance were selected. qRT-PCR confirmed that the expression levels of ZmMPK5, ZmNS2 and other genes were significantly different after stress in near-isogenic lines. 【Conclusion】 ZmCCT10 is a candidate gene involved in low nitrogen tolerance in maize and it participates in the low-nitrogen tolerance response of maize as transcriptional regulation.

Key words: maize, ZmCCT10, low nitrogen resistance, transcription factor, transcriptome

Table 1

qRT-PCR primers"

引物名称Primer name 序列Sequence (5′-3′)
ZmCCT10_qRTPCR-F AACGACGACGACCTCATCAG
ZmCCT10_qRTPCR-R ACTGGAACTCGTGCAGGCAC
ZmMPK5_qRTPCR-F ACTGATGGACCGCAAACC
ZmMPK5_qRTPCR-R GGGTGACGAGGAAGTTGG
Zmb-ZIP44_qRTPCR-F TCGATCGTTAACCAGCACGG
Zmb-ZIP44_qRTPCR-R GTTCGACTCCTTGCGCTTG
ZmNS3_qRTPCR-F CTGTTCACGGACCTCGTCAC
ZmNS3_qRTPCR-R GGTAGTTGCCGAAGTAGGGG
ZmNS2_qRTPCR-F TGCTTGCCGTGTACCACC
ZmNS2_qRTPCR-R CTTTTGGTGGAGCGTGTTCG
GAPDH-F ATCAACGGCTTCGGAAGGAT
GAPDH-R CCGTGGACGGTGTCGTACTT

Fig. 1

Y331-ΔTE and Y331 plants and roots after one week of control and low nitrogen stress treatment a: Y331-ΔTE and Y331 plants. Control: CK, LNS: Low nitrogen stress; b: Roots of Y331-ΔTE; c: Roots of Y331. 1: Primary radicle; 2: Main root; Except 1 and 2 from total root is lateral root"

Fig. 2

Traits analysis of low nitrogen stress of Y331-ΔTE and Y331 a: Total root length; b: Primary radicle length; c: Main root length; d: Lateral root length; e: Root dry weight; f: Shoot dry weight; g: SPAD value; h: Nitrogen accumulation; i: NR activity"

Fig. 3

Expression analysis of ZmCCT10 in leaves (a, c) and roots (b, d) of Y331-ΔTE and Y331 under low nitrogen stress"

Fig. 4

Identification and enrichment analysis of DEGs under low nitrogen stress a: Person correlation between samples; b: Venn diagram of DEGs; c: Cluster/heatmap of DEGs; d: Bar chart of GO enrichment analysis"

Fig. 5

qRT-PCR verification results of DEGs"

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