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Theoretical and Applied Genetics

, Volume 132, Issue 1, pp 113–123 | Cite as

A mutation in class III homeodomain-leucine zipper (HD-ZIP III) transcription factor results in curly leaf (cul) in cucumber (Cucumis sativus L.)

  • Fuxi Rong
  • Feifan Chen
  • Li Huang
  • Jiayu Zhang
  • Chaowen Zhang
  • Dong Hou
  • Zhihui Cheng
  • Yiqun Weng
  • Peng ChenEmail author
  • Yuhong LiEmail author
Original Article
  • 575 Downloads

Abstract

Key message

We identified two curly-leaf (cul) mutants in cucumber. Map-based cloning revealed that both mutants are due to allelic mutations in the CsPHB gene, a homolog of the Arabidopsis PHABULOSA which encodes a class III homeodomain-leucine zipper (HD-ZIP III) transcription factor.

Abstract

Leaf rolling is an important agronomic trait in crop breeding. Moderate leaf rolling minimizes shadowing between leaves, leading to improved photosynthetic efficiency. Although a number of genes controlling rolled leaf have been identified from rice and other plant species, none have been mapped or cloned in cucurbit crops. In this study, we identified and characterized two curly leaf (cul) mutants, cul-1 and cul-2 in cucumber. With map-based cloning, we show that cul-1 and cul-2 are allelic mutations and CsPHB (Csa6G525430) was the candidate gene for both mutants. The CsPHB gene encoded a class III homeodomain-leucine zipper (HD-ZIP III) transcription factor. A single non-synonymous mutation in the fourth and fifth exons of the CsPHB was responsible for the cul-1 and cul-2 mutant phenotypes, respectively. The single-nucleotide substitutions in cul-1 and cul-2 were both located in cs-miRNA165/166 complementary sites of CsPHB. The expression level of CsPHB gene in multiple organs of cul-1 and cul-2 mutants was higher than that in the wild type, while the expression of cs-miRNA165/166 in the two genotypes showed the opposite trend. We speculate that disruption of the binding between the mutant allele of CsPHB and cs-miRNA165/166 leads to the curly-leaf phenotype. This is the first report to clone and characterize the CsPHB gene in the family Cucurbitaceae. Taken together, these results support CsPHB as an important player in the modulation of leaf shape development in cucumber.

Notes

Acknowledgements

Work in YL’s laboratory was supported by the National Natural Science Foundation of China (31471891and 31772300). Work in YW’s laboratory was supported by the Agriculture and Food Research Initiative competitive grant no. 2017-67013-26195 of the USDA National Institute of Food and Agriculture. Work in ZC’s laboratory was supported by the National Key R&D Project (2016YFD0101705). Work in DH’s laboratory was supported by the National Natural Science Foundation of China (31860557).

Author contribution statement

FR performed the research and prepared a draft of the manuscript. FC, LH, CZ and JZ participated in the research. ZC and DH provided technical help. YW participated in data analysis and manuscript writing. PC and YL designed the experiments, supervised this study and wrote the manuscript. All authors have read and approved the manuscript.

Compliance with ethical standards

Conflict of interest

On behalf of all authors, the corresponding author states that there is no conflict of interest.

Supplementary material

122_2018_3198_MOESM1_ESM.pptx (1.7 mb)
Fig. S1 Morphological characterization of the leaves from the wild type (CCMC/wt, A) and the mutant (C1561/cul-1, B) (PPTX 1730 kb)
122_2018_3198_MOESM2_ESM.pptx (2.8 mb)
Fig. S2 Floral morphological characterization from wild-type (CCMC) and mutant (C1561/cul-1 and C1134/cul-2) plants. A. Heterozygote mutant (C1134/cul-2), female flower (A1, A2), male flower (A3) (bar = 1 cm). B. Homozygote mutant (C1561/cul-1), female flower (B1, B2), male flower (B3) (bar = 1 cm). C. Heterozygote mutant (C1561/cul-1), female flower (C1, C2), male flower (C3) (bar = 1 cm). D. Wild type (CCMC), female flower (D1, D2), male flower (D3) (bar = 1 cm) (PPTX 2846 kb)
122_2018_3198_MOESM3_ESM.pptx (708 kb)
Fig. S3 Morphological characterization of leaves and petioles from heterozygote mutant C1561/cul-1 (A), C1134/cul-2 (B) and wild-type CCMC (C) (bar = 1 cm) (PPTX 708 kb)
122_2018_3198_MOESM4_ESM.pptx (2.3 mb)
Fig. S4 Morphological characterization of wild-type and curly-leaf mutant plants in cucumber. A. Curly-leaf mutant (C1561/cul-1), heterozygote. B. Curly-leaf mutant (C1561/cul-1), homozygote. C. Curly-leaf mutant (C1134/cul-2), heterozygote. D. Wild type (bar = 8 cm) (PPTX 2315 kb)
122_2018_3198_MOESM5_ESM.pptx (65 kb)
Fig. S5 Genetic mapping of cul-1 and cul-2 loci in cucumber. A. Preliminary mapping with 94 F2 plants of C1561 × 9930 placed cul-1 in Chromosome 6, which was narrowed down to a region of 222 kb through fine mapping (B) with 1135 C1561 × 9930 F2 plants. C. Preliminary mapping with 240 F2 of C1134 × Gy14 also located cul-2 at cucumber Chromosome 6 (PPTX 65 kb)
122_2018_3198_MOESM6_ESM.pptx (5.1 mb)
Fig. S6 CAPS assay of natural cucumber populations with CAPS-C1561-01 derived from the causal SNP (SNP28494600) within the CsPHB candidate gene (PPTX 5253 kb)
122_2018_3198_MOESM7_ESM.pptx (3.9 mb)
Fig. S7 CAPS assay of natural cucumber populations with dCAPS-C1134-01 derived from the causal SNP (SNP28494713) within the CsPHB candidate gene (PPTX 3996 kb)
122_2018_3198_MOESM8_ESM.pptx (34 kb)
Fig. S8 Domain structure of CsPHB which is typically comprising of the homeobox, bzip, start and mekhla domains. Amino acid substitution from G in wild type to E in the C1134 mutant (a) and from P in wild type to L in the C1561 mutant (b) (PPTX 33 kb)
122_2018_3198_MOESM9_ESM.xlsx (32 kb)
Supplementary material 9 (XLSX 31 kb)
122_2018_3198_MOESM10_ESM.pdf (470 kb)
Supplementary material 10 (PDF 469 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Fuxi Rong
    • 1
  • Feifan Chen
    • 1
  • Li Huang
    • 1
  • Jiayu Zhang
    • 1
  • Chaowen Zhang
    • 1
  • Dong Hou
    • 2
  • Zhihui Cheng
    • 1
  • Yiqun Weng
    • 3
    • 4
  • Peng Chen
    • 5
    Email author
  • Yuhong Li
    • 1
    Email author
  1. 1.College of HorticultureNorthwest A&F UniversityYanglingChina
  2. 2.Vegetable Research InstituteGansu Academy of Agricultural SciencesLanzhouChina
  3. 3.Horticulture DepartmentUniversity of WisconsinMadisonUSA
  4. 4.Vegetable Crops Research UnitUSDA-ARSMadisonUSA
  5. 5.College of Life ScienceNorthwest A&F UniversityYanglingChina

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