Scientia Agricultura Sinica ›› 2023, Vol. 56 ›› Issue (16): 3051-3061.doi: 10.3864/j.issn.0578-1752.2023.16.001

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

Function of Maize ZCN7 in Regulating Drought Resistance at Flowering Stage

LI Yan1(), TAO KeYu2(), HU Yue3, LI YongXiang3, ZHANG DengFeng3, LI ChunHui3, HE GuanHua3, SONG YanChun3, SHI YunSu3, LI Yu3, WANG TianYu3, ZOU HuaWen1(), LIU XuYang3()   

  1. 1 College of Agriculture, Yangtze University, Jingzhou 434000, Hubei
    2 College of Agriculture Resources & Environment, Heilongjiang University, Harbin 150080
    3 Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081
  • Received:2023-03-09 Accepted:2023-05-18 Online:2023-08-16 Published:2023-08-18

Abstract:

【Objective】The main producing areas of maize is mostly located on the arid or semi-arid region that relying on the rainfed farming in China. The maize production losses caused by drought is a great threaten to food security. As a cross-pollinating crop, maize is mostly sensitive to water stress during flowering time. Drought at flowering stage will lead to asynchronous development between the male and female flower and cause massive grain yield loss. Thus, mining drought resistance related genes at flowering stage is important for maize drought resistance improvement and breeding. 【Method】In the present study, the phylogenic tree of 24 ZCN genes in maize genome, which is homologs of Arabidopsis FT gene, was build. The gene expression patterns of ZCN7 were analysis using qRT-PCR and in vivo GFP fluorescence imaging. A maize natural population consisting of 118 diverse inbred lines were planted in three environments, Beijing in 2021 and 2022 and Urumqi in 2022, to identify the flowering time related traits under different water treatments. The genomic variants around ZCN7 were detected by PCR and Sanger sequencing. The candidate gene association analysis was performed based on mixed linear model and the significant associated variants with drought induced anthesis-silking interval was obtained. The gene expression level of ZCN7 in natural population at flowering time was also measured by qRT-PCR. The differences of drought resistance traits and ZCN7 expression were compared between different haplotypes of significant associated variant. The Ubi1:ZCN7 overexpression transgenic maize were obtained, and the phenotypic performance was identified under different water treatments. 【Result】The 24 ZCN genes in maize genome included 15 FT like genes, 6 TFL1 like genes and 3 MFT like genes. The protein sequence of ZCN genes varied from 111 nn to 193 nn. The ZCN7 showed close relationship with ZCN8 and the protein sequence identity was 83.3% between the two genes. ZCN7 showed highest gene expression in the leaf blade at V12 stage. And the ZCN7-promoter:GFP vector was transformed to Arabidopsis and the GFP showed enriched signal at the blade edge of mature leaf. The candidate gene association analysis revealed a SNP variant at 1001 bp upstream of ZCN7 start codon had highest association signal with drought induced anthesis-silking interval under drought. The A/A and G/G haplotypes of SNP-1001 included 78 and 27 inbred lines, respectively. The anthesis-silking interval of A/A haplotype lines were significantly lower than G/G lines. And the ZCN7 gene expression of A/A haplotype lines were significantly higher than G/G lines. In addition, the ZCN7 overexpression transgenic lines showed significantly decreased anthesis-silking interval than wild type lines. Under drought, the anthesis-silking intervals of OE1 and OE2 were 2.3 and 2.6 days shorter than wild type lines. And the grain yield per plant and kernel number per plant of transgenic lines were significantly higher than wild type lines under drought, while the hundred kernel weight, kernel length and kernel width showed no significant difference. 【Conclusion】The maize ZCN7 played positive role in drought resistance and its overexpression improved grain yield by reducing anthesis-silking interval under drought.

Key words: maize (Zea mays L.), drought resistance, flowering time, ZCN

Fig. 1

The maize ZCN gene family The left panel is phylogenic tree of 24 ZCN genes in maize. The middle panel is the conserved motif of ZCN proteins. Boxes in different colors present conserved motifs. The right panel is gene structure of ZCN genes. The green boxes are UTR and red boxes are coding sequence"

Fig. 2

The gene expression patterns of ZCN7 a: The gene expression level of ZCN7 and ZCN8 in different tissues; b: The in vivo GFP fluorescence image"

Fig. 3

The candidate gene association analysis of ZCN7 a: The candidate gene association analysis of ZCN7 with anthesis-silking interval based on mixed linear model. The zero point of X axis is the start codon (ATG) of ZCN7. The gene structure of ZCN7 is plotted. The white boxes, black boxes and lines are UTR, coding sequence and introns. b-e: The anthesis-silking interval (ASI), days to anthesis (DTA), days to silking (DTS), and ZCN7 gene expression between different haplotypes of SNP-1001 under drought. f-i: The anthesis-silking interval (ASI), days to anthesis (DTA), days to silking (DTS), and ZCN7 gene expression between different haplotypes of SNP-1001 under well-watered conditions. The statistical analysis is performed by Students’ t-tes"

Fig. 4

The flowering time traits of ZCN7 overexpression transgenic maize The morphology of ZCN7 overexpression transgenic lines (OE1 and OE2) and wild type lines (WT) under drought (a) and well-watered (b) conditions. The ASI (c), DTA (d), and DTS (e) of transgenic and wild type lines under drought. The ASI (f), DTA (g), and DTS (h) of transgenic and wild type lines under well-watered conditions. The statistical analysis is performed by Students’ t-test"

Fig. 5

The grain yield traits of ZCN7 overexpression transgenic maize The grain yield per plant (a), hundred kernel weight (b), kernel length (c), kernel width (d) and kernel number per plant (e) of transgenic (OE1 and OE2) and wild type (WT) lines under drought. The grain yield per plant (f), hundred kernel weight (g), kernel length (h), kernel width (i), and kernel number per plant (j) of transgenic and wild type lines under drought. The statistical analysis is performed by Students’ t-test"

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