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Acta Agronomica Sinica ›› 2020, Vol. 46 ›› Issue (9): 1359-1367.doi: 10.3724/SP.J.1006.2020.03005


Phenotype identification and gene mapping of defective kernel 48 mutant (dek48) in maize

SHI Hui-Min1(), JIANG Cheng-Gong1,2, WANG Hong-Wu1, MA Qing2, LI Kun1, LIU Zhi-Fang1, WU Yu-Jin1, LI Shu-Qiang1, HU Xiao-Jiao1,*(), HUANG Chang-Ling1,*()   

  1. 1 Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, National Engineer Laboratory of Crop Molecular Breeding, Beijing 100081, China
    2 National Engineering Laboratory of Crop Stress Resistance Breeding / School of Life Sciences, Anhui Agricultural University, Hefei 230036, Anhui, China
  • Received:2020-01-20 Accepted:2020-04-15 Online:2020-09-12 Published:2020-04-26
  • Contact: Xiao-Jiao HU,Chang-Ling HUANG E-mail:3041601854@qq.com;huxiaojiao@caas.cn;huangchangling@caas.cn
  • Supported by:
    National Natural Science Foundation of China(31500984);Agricultural Science and Technology Innovation Program at CAAS;Central Public-interest Scientific Institution Basal Research Fund(Y2019CG15)


Maize kernel is not only a main nutrient storage organ, but also a model organ for seed development research of gramineous plants. In this study, a stable defective kernel mutant 48 (dek48) was identified from a library of mutants of the maize inbred line Zheng58 treated with ethylmethane sulfonate (EMS). Compared with wild type, the hundred-kernel weight of the dek48 was decreased greatly due to shrunken appearance and small flat size. Moreover, the dek48 was incapable of growing into a plantlet owning to the severely defective embryo and endosperm. The obvious defective development of the mutant can be observed at 12 days after pollination (DAP), indicating that the mutation occurred at an early stage of kernel development. Microscopic observation by scanning electron microscopy (SEM) revealed that starch granule of the dek48 was significantly smaller than wild type (WT). The observation of the paraffin section demonstrated that the starch granule of dek48 endosperm was partially filled and the aleurone layer cells developed irregularly. Genetic analysis based on kernel form indicated that the mutant trait was controlled by a single recessive gene. Based on genetic F2 population mapping, the gene of the mutant was located between 7.39 Mb-7.52 Mb on chromosome 3. The bioinformation analysis indicated that there were six new open reading frames (ORFs) and unknow genes related to kernel development in this region. Furthermore, candidate gene will be identified through sequencing and gene expression analysis in the future.

Key words: maize, defective kernel mutant, dek48, gene mapping

Table 1

Primer sequences used for gene mapping"

Forward sequence (5′-3′)
Reverse sequence (5′-3′)

Fig. 1

Phenotype of dek48 mutant in maize A: mature M4 ear; B: longitudinal section of wild type kernel; C: longitudinal section of dek48 kernel; D: F2 ear with B73 as the maternal parent; E: F2 ear with Mo17 as the maternal parent; F: F2 ear with C7-2 as the maternal parent.、"

Fig. 2

Hundred-kernel weight of wild type and mutant kernels ** Significant difference at P< 0.01."

Fig. 3

Determination and analysis of WT and dek48 kernel components A: starch content of WT and dek48; B: protein and oil content of WT and dek48. *, ** represent significant difference between the dek48 mutant and WT at the 0.05 and 0.01 probability levels, respectively."

Fig. 4

SEM observation of the endosperm of WT and dek48 A: SEM observation of WT, bar = 10 μm; B: SEM observation of dek48, bar = 10 μm; SG: starch granule; MP: matrix protein."

Fig. 5

Observation of embryo and endosperm of WT and dek48 at different development stages A: the observation and analysis of the embryo, endosperm and whole seed of dek48 at 12, 16, 20, 25, and 30 DAP, bar = 2 mm; B: the observation and analysis of the embryo, endosperm and whole seed of WT at 12, 16, 20, 25, and 30 DAP, bar = 2 mm."

Fig. 6

Paraffin section observation of WT and dek48 kernels at 16DAP A: embryos of WT seeds at 16 DAP, bar = 200 μm; B: embryo of dek48 seeds at 16 DAP, bar = 200 μm; C: aleurone layer of WT seeds at 16 DAP, bar = 50 μm; D: aleurone layer of dek48 seeds at 16 DAP, bar = 50 μm; AL: aleurone."

Table 2

Statistics and analysis of the segregation ratios of WT and dek48 mutants"

Total kernel number
M4 515 108/407 1.20
F2 (B73×dek48) 1019 236/783 1.84
F2 (C7-2×dek48) 1067 259/808 0.30
F2 (Mo17×dek48) 1401 326/1075 2.24

Fig. 7

Number of SNP markers"

Fig. 8

Distribution of SNP-index on whole genome"

Fig. 9

The fine mapping of dek48 mutant N: number of population; Recombinant: number of recombinants."

Table 3

Candidate genes annotation in the location interval"

Gene locus
Gene location
Gene annotation
Zm00001d039532 Chr. 3: 7,399,394-7,401,753 WRKY-transcription factor 56
Zm00001d039533 Chr. 3: 7,446,385-7,450,083 Plant cysteine oxidase 2
Zm00001d039534 Chr. 3: 7,451,813-7,457,371 F-box family protein
Zm00001d039535 Chr. 3: 7,479,692-7,482,979 HXXXD-type acyl-transferase family protein
Zm00001d039536 Chr. 3: 7,484,018-7,486,445 Syntaxin22
Zm00001d039537 Chr. 3: 7,487,245-7,530,464 Protein ALWAYS EARLY 3
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