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Acta Agronomica Sinica ›› 2018, Vol. 44 ›› Issue (10): 1433-1441.doi: 10.3724/SP.J.1006.2018.01433


Cloning and Function Analysis of ZmNAOD Gene in Maize

Chen-Yu MA,Wei-Min ZHAN,Wen-Liang LI,Meng-Di ZHANG,Zhang-Ying XI()   

  1. Agronomy College, Henan Agricultural University, Zhengzhou 450046, Henan, China
  • Received:2018-03-13 Accepted:2018-06-12 Online:2018-10-10 Published:2018-07-02
  • Contact: Zhang-Ying XI E-mail:xizhangying@163.com
  • Supported by:
    This study was supported by the National Natural Science Foundation of China(31371629)


The development of kernels and the photoperiod characters are key elements that impact yield in maize. A N-acetylornithine deacctylase (NAOD) gene named ZmNAOD was cloned from maize inbred line Chang 7-2 by the method of RT-PCR in our work. The length of CDS (coding DNA sequence) in ZmNAOD gene is 1344 bp, coding a polypeptide of 447 amino acids. The expression of ZmNAOD was abundant in tassel, kernel, leaf, stem and root successively, and was up-regulated in seed from 0 to 15 d, and down-regulated after 15 d. The over-expression recombinant plasmid, pCAMBIA1304-ZmNAOD, was transformed into Arabidopsis using Agrobacterium mediated method, and homogeneous transgenic lines were obtained. The ZmNAOD gene expression was abundant in roots of transgenic Arabidopsis, and the root length was significantly longer than that of the wild type (WT) after a 10-day dark treatment. Flowering days of transgenic lines were earlier, the seeds were longer and the thousand seed weight was heavier than those of the WT. These results demonstrated that the ZmNAOD might be involved in seed development and photoperiod.

Key words: maize, gene cloning, genetic modification, seed development, photoperiod

Fig. 1

DNA coding sequence and amino acid alignments of ZmNAOD"

Fig. 2

Phylogenetic analysis of NAOD proteins in plants"

Table 1

Prediction cis-elements of ZmNAOD promoter with database analysis"

Element (core sequence)
胚乳表达元件 cis-acting regulatory element required for endosperm expression Skn-1_motif (GTCAT), GCN4_motif (TGTGTCA)
昼夜节律控制元件 cis-acting regulatory element involved in circadian control Circadian (CAANNNNATC)
光响应相关元件 cis-acting regulatory element involved in light responsiveness G-box (CACGTC), Sp1 (CC(G/A)CCC)
与光响应相关保守 DNA模块
Part of a conserved DNA module involved in light responsiveness
部分光响应相关元件 Part of a light responsive element

脱落酸响应元件 cis-acting element involved in the abscisic acid responsiveness ABRE (AGTACGTGGC)
cis-acting regulatory element involved in auxin responsiveness
AuxRR-core (GGTCCAT)
cis-acting regulatory element involved in the MeJA-responsiveness
CGTCA-motif (CGTCA), TGACG-motif (TGACG)
水杨酸响应元件 cis-acting element involved in salicylic acid responsiveness TCA-element (TCAGAAGAGG)
热响应相关元件 cis-acting element involved in heat stress responsiveness HSE (AAAAAATTTC)
低温响应元件 cis-acting element involved in low-temperature responsiveness LTR (CCGAAA)
MYB结合位点 MYB binding site involved in drought-inducibility MBS (TAACTG)
逆境响应元件 cis-acting element involved in defense and stress responsiveness TC-rich repeats (ATTTTCTTCA)

Fig. 3

Relative expression of ZmNAOD gene in different organs in Chang 7-2"

Fig. 4

Relative expression of ZmNAOD gene in kernel at different days after pollination"

Fig. 5

Relative expression of ZmNAOD gene in wild type (WT) and transgenic lines"

Fig. 6

Relative expression of ZmNAOD gene in different organs of transgenic plant"

Table 2

Flowering time in wild type and transgenic lines"

野生型 Wild type T3-17 T3-23 T3-26
第1朵可见花序的天数 Days of the first buds visible (d) 37.60±0.47 36.08±0.42* 34.80±1.08** 36.00±0.53*
第1朵花开花的天数 Days of the first flower visible (d) 48.23±0.55 46.96±0.64* 43.00±0.72** 46.00±0.65**

Fig. 7

Flowering phenotype observed in transgenic lines and wild type lines at 45 days after sowing 1, 2: transgenic lines T3-23; 3, 4: wild type lines."

Table 3

Analysis of seed phenotype of wild type and transgenic lines"

Wild type
Transgenic T3-17
Transgenic T3-23
Transgenic T3-26
粒长 Seed length (mm) 0.7857±0.0380 0.8936±0.0449** 0.9004±0.0527** 0.9029±0.0564**
粒宽 Seed width (mm) 0.6682±0.0259 0.6798±0.0128 0.6656±0.0315 0.6646±0.0220
千粒重 Thousand-seed weight (mg) 13.353±0.122 14.233±0.031** 14.249±0.137** 14.222±0.152**
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