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Acta Agronomica Sinica ›› 2019, Vol. 45 ›› Issue (5): 656-661.doi: 10.3724/SP.J.1006.2019.83058

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

Genetic analysis and causal gene identification of maize viviparous mutant vp-like8

Rui WANG1,Yang-Song CHEN1,Ming-Hao SUN1,2,Xiu-Yan ZHANG3,Yi-Cong DU1,Jun ZHENG1,*()   

  1. 1 Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China
    2 College of Agronomy, Jilin Agricultural University, Changchun 130118, Jilin, China
    3 School of Life Science, China Agricultural University, Beijing 100193, China
  • Received:2018-08-15 Accepted:2019-01-12 Online:2019-05-12 Published:2019-02-22
  • Contact: Jun ZHENG E-mail:zhengjun02@caas.cn
  • Supported by:
    This work was supported by the National Key Research and Development Program of China(2016YFD0101002);the Agricultural Science and Technology Innovation Program of Chinese Academy of Agricultural Sciences.

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Abstract:

The maize mutant vp-like8 shows clear viviparous phenotype and stable inheritance, and genetic analysis showed that the mutant phenotype was controlled by a single recessive gene. Using an F2 segregation population derived from vp-like8 and inbred line Zheng 58, the causal gene was mapped to an interval from 160.4 Mb to 165.6 Mb on chromosome 3 by the BSR-Seq technology. According to the maize genomic database, a previously discovered viviparous gene Vp1 was identified to be in this mapping interval. The test crosses from vp1 and vp-like8 heterozygous plants showed a 3:1 segregation ratio between normal and viviparous kernels. The genomic sequence analysis revealed that vp-like8 mutant had a 343 bp deletion in the second intron and 222 bp insertion in the third intron of Vp1 gene, which is different from vp1 mutation of an only 343 bp deletion in the second intron of Vp1 gene. Further real time PCR analysis revealed that, compared with the normal kernels, the transcript level of Vp1 was significantly decreased both in vp-like8 and vp1 viviparous kernels. Taken together, these evidences suggest that vp-like8 is a new allele mutant of Vp1.

Key words: maize, viviparous, mutant, Vp1, gene mapping

Table 1

Primers used in this study"

引物
Primer		 正向序列
Forward sequence (5°-3°)		 反向序列
Reverse sequence (5°-3°)
VP1-G-F1/R1 GCGAGACCTGAAAACACACA CATGGCGTTCTCTAGCATCA
VP1-G-F2/R2 CGCACTCCCAAGAGAACC ATAGGGTAAGAGCCCGTGGA
VP1-G-F3/R3 GTGGTCGTGAACAGCCAAC GCTCTGCTTCAGCACCTTCT
VP1-G-F4/R4 CATCGCTGTCGAGCAATAGA CTGTACCGCATGTTCCACAC
VP1-G-F5/R5 TAAAATCGGCCATGGATAGG TCTCTGGCCCAGTGGTTAGT
VP1-G-F6/R6 GCTGCTGTTTTCCTCGAATC GCACCTAGCTGCCAAACACT
VP1-G-F7/R7 AGTCCTCCGGATCTCTCGTT AAACGGTTGCGTAGATTTGG
VP1-G-F8/R8 CCAGTGCAATGTCAGTGCTT AATGGCCGAGAGATCAGGTA
VP1-CDS-F1/R1 AGAAGGTGCTGAAGCAGAGC AACGAACAAATTCCCCTGTG
Zm-GAPDH-F/R CCCTTCATCACCACGGACTAC AACCTTCTTGGCACCACCCT

Fig. 1

Viviparous phenotype of vp-like8 mutant A: viviparous and normal kernels on a vp-like8 heterozygous ear at 30 days after self-pollination; B: viviparous and normal kernels on a vp-like8 heterozygous ear at 60 days after self-pollination; C: mature normal (WT) and viviparous kernels (vp-like8); Bar = 1 cm."

Table 2

Segregation of normal and viviparous kernels on vp-like8 self-pollinated heterozygous ears"

年度
Year		 地点
Location		 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)
2014 海南Hainan vp-like8/+ 150 45 195 0.289
361 118 479 0.017
2016 北京Beijing vp-like8/+ 132 46 178 0.030
121 38 159 0.052

Fig. 2

Gene mapping of the vp-like8 mutant by the BSR-Seq strategy"

Fig. 3

Allelism test of vp-like8 with vp1 by heterozygous mutants A: viviparous kernels were emerged on vp-like8 heterozygous ear crossed by the mixed pollen of vp1 heterozygous plants; B: viviparous kernels were emerged on vp1 heterozygous ear crossed by the mixed pollen of vp-like8 heterozygous plants."

Fig. 4

Gene structure of Vp1 and mutation site of two mutants"

Table 3

Test of vp-like8 with vp1"

父母本基因型
Parental genotype		 籽粒表型 Kernel phenotype
正常籽粒
Normal		 穗发芽籽粒
Viviparous		 总数
Total		 χ2
(3:1)
vp-like8/+ × vp1 /+ 208 72 280 0.042
vp1 /+ × vp-like8/+ 158 48 206 0.233

Fig. 5

Gene expression level of Vp1 in normal and viviparous (vp) kernels of vp-like8 and vp1 mutants by quantitative real- time PCR analysis"

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