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Identification and mapping of Tril, a homeodomain-leucine zipper gene involved in multicellular trichome initiation in Cucumis sativus

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Using map-based cloning of Tril gene, we identified a homeodomain-leucine zipper gene involved in the initiation of multicellular trichomes (including the spines of fruit) in cucumber.

Abstract

Fruit spines are a special type of trichome that impacts the quality and appearance of cucumber (Cucumis sativus L.) fruit. Scanning electron microscopy revealed that the trichome-less (tril) mutant originating from European greenhouse cucumber has a completely glabrous phenotype on cotyledons, hypocotyls, young leaves, fruits, and fruit stalks. Genetic analysis revealed that tril was inherited as a recessive allele at a single locus. Using 1058 F2 individuals derived from a cross between cucumber tril mutant CGN19839 and the micro-trichome (mict) mutant 06-2, tril was mapped to chromosome 6, and narrowed down to a 37.4 kb genomic region which carries seven predicted genes. Genetic and molecular analyses revealed that gene Cucsa.045360 is a possible candidate gene for the differentiation of epidermal cells to trichomes. It is a member of the class IV homeodomain-leucine zipper (HD-Zip IV) family and encodes homeodomain and START domain, sharing 66.7 % predicted amino acid sequence identity to PROTODERMAL FACTOR2 (PDF2) and 35.0 % to GLABRA2 (GL2) of Arabidopsis. The homeobox domain had changed amino acid sequence because of an insertion in tril mutant. The results of genetic analysis and transcriptome profiling indicated that the Tril gene had an epistatic effect on the Mict gene in trichome development. Phenotypes of the tril mutant such as glabrous fruits and female flowers at every node could be used in developing new cultivars.

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Acknowledgments

We thank Dr. Yi-qun Weng (Horticulture Department, University of Wisconsin-Madison, USA) for affording information of cucumber genetic map. This work was supported by grants from the National Natural Science Foundation of China (No. 31471156), China 973 Program (No. 2012CB113900), the Science and Technology Commission of Shanghai Municipality (No. 13JC1403600), the Shanghai Graduate Education and Innovation Program (Horticulture), the China Innovative Research Team, Ministry of Education.

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Authors and Affiliations

  1. School of Agriculture and Biology, Shanghai Jiao Tong University, 800 Dongchuan Road, Minhang District, Shanghai, 200240, China

    Yun-Li Wang, Jing-tao Nie, Chun-li Guo, Jian Pan, Huan-Le He, Jun-Song Pan & Run Cai

  2. Hunan Vegetable Research Institute, Hunan Academy of Agriculture Sciences, Changsha, 410125, China

    Hui-Ming Chen

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  1. Yun-Li Wang
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Correspondence to Jun-Song Pan or Run Cai.

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The experiments comply with the current laws of the country in which we were performed.

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The authors declare that they have no conflict of interest.

Additional information

Communicated by M. J. Havey.

Electronic supplementary material

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122_2015_2628_MOESM1_ESM.doc

Fig. S1 The tril cucumber plants show other phenotype. Besides glabrous phenotype, the tril mutant plants show unbranched (a), gynoecious (b, upper red arrow), leaf curvature (b, lower red arrow), and petals opened 1-2 days before flowering (c). (DOC 6235 kb)

122_2015_2628_MOESM2_ESM.doc

Fig. S2 Amino-acids sequence of six predicted genes. A comparison of the tril mutant with non-glabrous type Gy14, 9930, and 06-2 plants. Shaded residues indicate strong conservation. There were no differences between the tril mutant and non-glabrous type plants in the deduced amino acid sequences of these six predicted genes, respectively. (a) Cucsa.045370. (b) Cucsa.045380. (c) Cucsa.045390. (d) Cucsa.045400. (e) Cucsa.045410. (f) Cucsa.045420. (DOC 83 kb)

122_2015_2628_MOESM3_ESM.doc

Fig. S3 Amino acid alignment of the predicted protein of Tril ( Cucsa.045360 ) with seven HD proteins from other species. The two conserved domains, an HD domain and a START domain, are represented by black lines. These two regions are highly conserved among their homologues. At: Arabidopsis thaliana, Cm: Cucumis melo, Cs: Cucumis sativus, Gh: Gossypium hirsutum, Ns: Nicotiana sylvestris, Rt: Populus trichocarpa, Rc: Ricinus communis, and Vv: Vitis vinifera. (DOC 85 kb)

122_2015_2628_MOESM4_ESM.doc

Fig. S4 Comparison of Cucsa.045360 gene structures in wild-type (a) and tril mutant (b). The predicted gene structure revealed that the Cucsa.045360 gene with an insertion should be separated to two transcripts in the tril mutant (b). The START domain still retained its original function in the tril mutant, while the homeobox domain was changed because of a fragment insertion. (DOC 69 kb)

122_2015_2628_MOESM5_ESM.doc

Fig. S5 Expression of Cucsa.045360 of wild-type 9930 and tril mutant using RT-PCR analyses. Polymorphism only appeared in the first part of the transcription between 9930 and tril mutant. M: Markers (DL10000). The sequences of primers used are given in Table S2. (DOC 32 kb)

Table S1. Primers of the six functional predicted genes used for polymorphism (DOC 51 kb)

Table S2. Primers used for RT-qPCR (DOC 35 kb)

Supplementary material 8 (DOC 40 kb)

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Wang, YL., Nie, Jt., Chen, HM. et al. Identification and mapping of Tril, a homeodomain-leucine zipper gene involved in multicellular trichome initiation in Cucumis sativus . Theor Appl Genet 129, 305–316 (2016). https://doi.org/10.1007/s00122-015-2628-4

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