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Acta Agronomica Sinica ›› 2024, Vol. 50 ›› Issue (11): 2712-2719.doi: 10.3724/SP.J.1006.2024.43008

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

Cloning and functional analysis of viviparous mutant vp2 in maize

ZHANG Xin-Yue1,2(), QIN Yang2, LI Rui3, HUANG Quan-Sheng4, WANG Yi-Ru2,*(), ZHENG Jun1,2,*()   

  1. 1College of Agronomy, Qingdao Agricultural University, Qingdao 266109, Shandong, China
    2Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China
    3College of Agriculture, Gansu Agricultural University, Lanzhou 730070, Gansu, China
    4Institute of Nuclear Technology and Biotechnology, Xinjiang Academy of Agricultural Sciences, Urumqi 830091, Xinjiang, China
  • Received:2024-02-05 Accepted:2024-06-20 Online:2024-11-12 Published:2024-07-11
  • Contact: *E-mail: zhengjun02@caas.cn; E-mail: wangyiru@caas.c
  • Supported by:
    Open Project of Key Laboratory, Xinjiang Uygur Autonomous Region(2023D04070)

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Abstract:

Maize vivipary, the precocious germination of seeds on the ear, significantly impacts maize yield and quality. Developing vivipary-resistant maize varieties through the discovery of novel genes is crucial for agricultural production in China. In this study, the maize mutant vp2 exhibited a clear viviparous phenotype with stable inheritance, controlled by a single recessive gene. Genome sequence analysis of the vp2 mutant revealed deletions in two coding genes (Zm00001d015355 and Zm00001d015356), with Zm00001d015356 encoding p-hydroxypyruvate dioxygenase (ZmHPPD1). The hppd1 mutant also displayed a viviparous phenotype. Furthermore, test crosses between vp2 and hppd1 heterozygous plants showed a 3:1 segregation ratio between normal and viviparous kernels, suggesting that ZmHPPD1 is the candidate gene for vp2. To further investigate the mechanism by which ZmHPPD1 regulates maize vivipary, we analyzed endogenous hormone and metabolite content in the ABA synthesis pathway. The results indicated a significant decrease in ABA levels, a substantial accumulation of octahydro lycopene, and a notable reduction in purple xanthophyll, zeaxanthin, and lutein in viviparous kernels. ZmHPPD1 disrupts ABA synthesis by affecting the conversion of octahydro lycopene to lycopene, leading to the loss of dormancy and early germination of maize kernels. These findings provide valuable genetic resources for breeding vivipary-resistant maize.

Key words: maize, viviparous, plant hormones, abscisic acid

Fig. 1

Viviparous phenotype of vp2 mutant A: ear and kernels phenotypes of the vp2 heterozygous mutant at 25 days after pollination, scale bar: 1 cm; B: longitudinal sections of normal kernels (WT) and viviparous kernels (vp2) harvested at 18 d, 21 d, and 25 d; bar: 500 μm."

Table 1

Primers used in this study"

引物
Primer		 引物序列
Primer sequences (5′-3′)
vp2-F1 CGCAGCCAGATACAAACGTT
vp2-R1 GGGACATGGGAGCTCGAATT
TIR8.1 GAAGCCAACGCCAWCGCCTCYATTTCGTCGAAT
vp2-F2 ACTACGGGCTGAGCAGGTT
vp2-R2 TGAAGATTTGGAGCAGCACG
GAPDH-F CCCTTCATCACCACGGACTAC
GAPDH-R AACCTTCTTGGCACCACCCT

Table 2

Segregation of normal and viviparous kernels on vp2 self-pollinated heterozygous ears"

植株基因型
Plant genotype		 籽粒表型 Kernel phenotype
正常籽粒
Normal		 穗发芽籽粒
Viviparous		 总数
Total		 χ2
(3:1)
B73×vp2/+ 162 48 210 0.406
128 37 165 0.454

Fig. 2

Vp2 encodes p-hydroxyphenylpyruvate dioxygenase ZmHPPD1 A: gene mapping of vp2 mutant genes by BSR-Seq; B: gene differences between B73 and vp2 mutant materials; C: expression of ZmHPPD1 in vp2 mutants."

Fig. 3

Allelic test of hppd1 with vp2 by heterozygous mutants A: gene structure of ZmHPPD1 and mutation site of hppd1; B: identification of Mu transposon insertion in hppd1; C: ear and kernels phenotypes of hppd1 heterozygous mutants; D: the ear and kernels phenotypes of the hybrid between hppd1 and vp2."

Table 3

Separation of normal and viviparous kernels on hppd1 self-pollinated heterozygous ears and the crosses of vp2 and hppd1 heterozygous ears"

植株基因型
Plant genotype		 籽粒表型 Kernel phenotype
正常籽粒
Normal		 穗发芽籽粒
Viviparous		 总数
Total		 χ2
(3:1)
hppd1/+ 96 31 127 0.003
128 44 172 0.008
hppd1/+×vp2/+ 171 64 235 0.512
156 57 213 0.264

Fig. 4

Analysis of endogenous hormone in the kernels of WT and vp2 A: ABA; B: ABA-GE; C: SA; D: IAA; E: SL; F: JA; G: CK; H: ETH. ** indicates significant correlation at the P < 0.01."

Fig. 5

Content of related metabolites in the ABA synthesis pathway A: (E/Z)-phytoene; B: α-carotene; C: lutein; D: β-carotene; E: violaxanthin; F: zeaxanthin. *, ** indicate significant correlation at the P < 0.05 and P < 0.01, respectively."

Fig. 6

ABA biosynthetic pathway of WT and vp2 A: the ABA biosynthesis pathway of WT; B: the ABA biosynthesis pathway of vp2 mutant. In the metabolic pathway, green indicates a decrease of metabolite, and red indicates an increase of metabolite."

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