Skip to main content
Log in

Chromosome doubling of pepper haploids via chemical or high-temperature treatment

  • Published:
Euphytica Aims and scope Submit manuscript

Abstract

In pepper haploid breeding, the low natural doubling rate is a major bottleneck. Artificially inducing haploids chromosome doubling can obtain pepper diploids and improve haploid breeding efficiency. While there is no effective way reported in pepper up to now. This study aimed to investigate the relationship between pepper microsporogenesis and flower development to guide diploid induction and also explore suitable conditions for treatments using chemical reagents and high temperature. The results showed that flower buds developed to stage 2 was most suitable for chemical and high-temperature treatment, when the corolla and anther were green, and the corolla slightly exposed the bracts, and the meiosis were in the zygotene–diplotene phase; the highest induction rate reached 30% when growth points of pepper haploids were treated with 0.006% pendimethalin for 2 days, which is higher than the traditional method of colchicine treatment; the highest induction rate for high-temperature treatment was 14.29%, when the flower buds were subjected to high-temperature treatment at 38 °C for 2 h. The study first report that high-temperature treatment is a feasible method for chromosome doubling of pepper haploids and provide an efficient and practical chemical treatment method for pepper chromosome doubling.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

Abbreviations

APM:

Amiprohpos methyl

DH:

Doubled haploid

HAP:

Hydroponics after pollination

PMC:

Pollen mother cell

References

  • Allum JF, Bringloe DH, Roberts AV (2007) Chrmosome doubling in a Rosa rugosa Thunb hybrid by exposure of in vitro no-desto oryzalin: the effect of nodelength, oryzalin concentration and exposure time. Plant Cell Rep 26(11):1977–1984

    Article  CAS  PubMed  Google Scholar 

  • Bosland PW, Votava EJ (2000) Peppers: vegetable and spice Capsicum. CABI Publishing, New York

    Google Scholar 

  • Bouvier L, Guerif P, Djulbic M, Durel CE, Chevreau E, Lespinasse Y (2002) Chromosome doubling of pear haploid plants and homozygosity assessment using isozyme and microsatellite markers. Euphytica 123:255–262

    Article  CAS  Google Scholar 

  • Bradshaw HD, Stettler RF (1993) Molecular genetics of growth and development in Populus. I. Triploidy in hybrid poplars. Theor Appl Genet 86(2–3):301–307

    Article  PubMed  Google Scholar 

  • Chakraborti SP, Vijayan K, Roy BN, Qadri SMH (1998) In vitro induction of tetraploidy in mulberry (Morus alba L). Plant Cell Rep 17(10):799–803

    Article  CAS  PubMed  Google Scholar 

  • Chen ZZ, Snyder S, Fan ZG, Loh WH (1994) Efficient production of doubled haploid plants through chromosome doubling of isolated mircrospores in Brassica napus. Plant Breeding 113:217–221

    Article  Google Scholar 

  • Djian-Caporalino C, Fazari A, Arguel MJ (2007) Root-knot nematode (Meloidogyne spp.) Me resistance genes in pepper (Capsicum annuum L.) are clustered on the P9 chromosome. Theor Appl Genet 114(3):473–486

    Article  CAS  PubMed  Google Scholar 

  • Doer C (1986) Evaluation ploidy level in plants of a population of brussels sprouts (Brassica oleareea L.ssp.gemmifera) derived from anther culture. Agronomie 6(9):797–801

    Google Scholar 

  • Ewald D, Ulrich K, Naujoks G, Schroeder MB (2009) Induction of tetraploid poplar and black locust plants using colchicine: chloroplast number as an early marker for selecting polyploids in vitro. Plant Cell Tiss Organ Cult 99(3):353–357

    Article  Google Scholar 

  • Fahleson J, Dixelius J, Sundberg E, Glimelius K (1988) Correlation between flow cytometric determination of nuclear DNA content and chromosome number in somatic hybrids within Brassicaceae. Plant Cell Rep 7(1):74–77

    Article  CAS  PubMed  Google Scholar 

  • Ferrie AMR, Caswell KL (2011) Isolated microspore culture techniques and recent progress for haploid and doubled haploid plant production. Plant Cell Tiss Organ Cult 104:301–309

    Article  Google Scholar 

  • Gao P, Lin W, Kang X (2004) Pollen chromosome doubling of Eucommia ulmoides induced by colchicine. J Beijing Univ 26:39–42

    Google Scholar 

  • Guo LQ, Xu WT, ZhangY ZJF, Wei Z (2017) Inducing triploids and tetraploids with high temperature in Populus sect Tacamahaca. Plant Cell Rep 36:313–326

    Article  CAS  PubMed  Google Scholar 

  • Hansen NJP, Andersen SB (1996) In vitro chromosome doubling potential of colchicine, oryzalin, trifluralin and APM in Brassica napus microspore culture. Euphytica 88(2):159–164

    Article  CAS  Google Scholar 

  • Harder LD, Thomson JD (1989) Evolutionary options for maximizing pollen dispersal of animal pollinated plants. Am Nat 133(3):323–344

    Article  Google Scholar 

  • Jia ZH, Zhao ZL, Xu M, Guo JW (2011) The correlation between microspore developmental stage and bud shape in linear pepper. J China Capsicum 2:24–27

    Google Scholar 

  • Kang XY (1996) Chromosome count and shape of poplar. J Gansu Agric Univ 31(1):67–70

    Google Scholar 

  • Kang XY, Zhu ZT, Lin HB (1999) Study on the effective treating period for pollen chromosome doubling of Populus tomentosa 9 P. bolleana. Sci Silvae Sin 35(4):21–24

    Google Scholar 

  • Kang XY, Zhu ZT, Zhang ZY (2000a) Breeding of triploids by the reciprocal crossing of Populus alba 9 P. glandulosa and P. tomentosa 9 P. bolleana. J Beijing Univ 22(6):8–11

    Google Scholar 

  • Kang XY, Zhu ZT, Zhang ZY (2000b) Suitable period of tomentosa × P. bolleana. J Beijing Univ 22:1–4

    Google Scholar 

  • Kang XY, Zhang PD, Gao P, Zhao F (2004) Discovery of a new way of poplar triploids induced with colchicine after pollination. J Beijing Univ 26(1):1–4

    Google Scholar 

  • Kulkarni M, Orse TB (2010) Induced polyploidy with gigas expression for root traits in Capsicum annuum L. Plant Breeding 129:461–464

    Google Scholar 

  • Li GP, Huang QC (2006) Ontogeny of pollen and pollination in keteleeria fortunei. Sci Silvae Sin 42(5):42–47

    Google Scholar 

  • Li SL, Lijuan X (2014) Induction of tetraploidy in non-heading Chinese cabbage (Brassica campestris ssp. chinensis Makino) by colchicine treatment increases the ascorbic acid concentration. J Horticult Sci Biotechnol 89(1):53–60

    Article  Google Scholar 

  • Liu G, Li Z, Bao M (2007) Colchicine-induced chromosome doubling in Platanus acerifolia and its effect on plant morphology. Euphytica 157:145–154

    Article  Google Scholar 

  • Lu M, Zhang PD, Kang XY (2013) Induction of 2n female gametes in Populus adenopoda Maxim by high temperature exposure during female gametophyte development. Breed Sci 63(1):96–103

    Article  PubMed  PubMed Central  Google Scholar 

  • Lu M, Zhang P, Wang J (2014) Induction of tetra ploidy using high temperature exposure during the first Zygote division in Populus adenopoda Maxim. Plant Growth Regul 72(3):279–287

    Article  CAS  Google Scholar 

  • Magnard JL, Yang M, Chen YCS, Leary M, McCormick S (2001) The Arabidopsis gene Tardy Asynchronous Meiosis is required for the normal pace and synchrony of cell division during male meiosis. Plant Physiol 127(3):1157–1166

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Mashkina OS, Burdaeva LM, Belozerova MM, Vyunova LN (1989) Method of obtaining diploid pollen of woody species. Lesoved 1:19–25

    Google Scholar 

  • Mityko J, Fari M (1997) Problems and results of doubled haploid plant production in pepper (capsicum annuum L.) via anther and microspore culture. Acta Hort 447:281–287

    Article  Google Scholar 

  • Moller C, Iqbal MCM, Robbelen G (1994) Efficient production of doubled haploid Brassica napus plants by colchicine treatment of microspores. Euphytica 75(1–2):95–104

    Article  Google Scholar 

  • Pozzobon MT, Vails JFM (2000) Cytogeography and variation of stomatal size of Paspalum glaucescens (Gramineae; Paniceae) in southern Brazil. Euphytica 116(3):251–256

    Article  Google Scholar 

  • Randolph LF (1932) Some effects of high temperature on polyploidy and other variations in maize. Proc Natl Acad Sci 18:222–229

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Risso-Pascotto C, Pagliarini MS, Do Valle CB, Jank L (2004) Asynchronous meiosis in an interspecific hybrid of Brachiaria ruziziensis and B brizantha. Plant Cell Rep 23(5):304–310

    Article  CAS  PubMed  Google Scholar 

  • Seguí-Simarro JM, Corral-Martínez P, Parra-Vega V, González-García B (2011) Androgenesis in recalcitrant solanaceous crops. Plant Cell Rep 30:765–778

    Article  PubMed  CAS  Google Scholar 

  • Sun QR, Sun HS, Li LG, Bell RL (2009) In vitro colchicine-induced polyploid plantlet production and regeneration from leaf explants of the diploid pear (Pyrus communis L.) cultivar ‘Fertility.’ J Hortic Sci Biotechnol 84:548–552

    Article  CAS  Google Scholar 

  • Tavoletti S (1994) Cytological mechanisms of 2n egg formation in a diploid genotype of Medicago sativa subsp. falcate. Euphytica 75(1–2):1–8

    Article  Google Scholar 

  • Tosca A, Pandoifi R, Citterio S, Fasoli A, Sgorbati S (1995) Determination by flow cytometry of the chromosome doubling capacity of colchicine and oryzalin in gynogenefic haploids of Gerbera. Plant Cell Rep 14:455–458

    Article  CAS  PubMed  Google Scholar 

  • Wan Y, Duncan DR, Rayburn AL, Petolion JF, Widholm JM (1991) The use of antimicrotubule herbicides for the production of doubled haploid plants from anther-derived maize callus. Thero Appl Genet 81(2):205–211

    Article  CAS  Google Scholar 

  • Wang J, Kang XY, Li DL, Jing YC, Zhang ZH (2006) Meiosis and chromosome behavior of pollen mother cell in Populus simonii. Acta Bot Boreali Occident Sin 26(11):2231–2238

    Google Scholar 

  • Wang J, Kang XY, Li DL (2008) Chromocenter number and relationship between it and ploidy level in Populus. Acta Bot Boreali Occident Sin 28(10):1968–1971

    Google Scholar 

  • Wang J, Kang XY, Li DL, Chen HW, Zhang PD (2010) Induction of diploid eggs with colchicine during embryo sac development in Populus. Silvae Genet 59(1):40–48

    Article  Google Scholar 

  • Wang J, Kang XY, Li DL (2012a) High temperature-induced triploid production during embryo sac development in Populus. Silvae Genet 61(3):85–93

    Article  Google Scholar 

  • Wang J, Li DL, Kang XY (2012b) Induction of unreduce megaspores with high temperature during megasporogenesis in Populus. Ann Forest Sci 15:1–9

    CAS  Google Scholar 

  • Wang J, Shi L, Song SY, Tian J, Kang XY (2013) Tetraploid production through zygotic chromosome doubling in Populus. Silva Fenn 47(2):1–12

    Article  Google Scholar 

  • Wei YX, Zhu MZ, Qiao HY, Li F, Zhang SJ, Zhang SF, Zhang H, Sun RF (2018) Characterization of interspecific hybrids between flowering Chinese cabbage and broccoli. Sci Hortic 240:552–557

    Article  Google Scholar 

  • Wyatt R (1982) Inflorescence architecture: how flower number, arrangement, and phenology affect pollination and fruit set. AmJ Bot 69(4):585–594

    Article  Google Scholar 

  • Xi XJ, Li D, Xu WT, Guo LQ, Zhang JF, Li BL (2012) 2n egg formation in Populus 9 euramericana (Dode) Guinier. Tree Genet Genomes 8(6):h1237-1245

    Article  Google Scholar 

  • Yang J, Yao PQ, Li Y, Mo JY (2016) Induction of 2n pollen with colchicine during microsporogenesis in Eucalyptus. Euphytica 210:69–78

    Article  CAS  Google Scholar 

  • Yang XM, Cao ZY, An LZ, Wang YM, Fang XW (2006) In vitro tetraploid induction via colchicine treatment from diploid somatic embryos in grapevine (Vitis vinifera L.). Euphytica 152(2):217–224

    Article  Google Scholar 

  • Zhao JP, Simmonds DH (1995) Application of trifluralin to embryogenic microspore cultures to generate doubled haploid plants in Brassica napus. Physiol Plant 95(2):304–309

    Article  CAS  Google Scholar 

  • Zhao JP, Simmonds DH, Newcomb W (1996) High frequency production of doubled haploid plants of Brassica napus cv Topas derived from colchicine-induced micropore embryogenesis without heat shock. Plant Cell Rep 15(9):668–671

    Article  CAS  PubMed  Google Scholar 

  • Zhang XF, Geng SS, Chen B (2009) Relationship between chromosome ploidy and stomatal characters of leaf in pepper. Jiangsu J Agr Sci 25:339–342

    Google Scholar 

  • Zhang XZ, Liu GJ, Yan LY, Zhao YB, Chang RF, Wu LP (2003) Creating triploid germplasm via induced 2n pollen in Capsicum annuum L. J Hortic Sci Biotechnol 78(1):84–88

    Article  Google Scholar 

Download references

Acknowledgements

This work was supported by General Program of National Natural Science Foundation of China (32072566), the Program of Beijing Municipal Science and Technology Committee (Z191100004019010), Innovation Ability Construction Project of Beijing Academy of Agriculture and Forest Sciences (KJCX20200113).

Author contribution statement

XFZ, SSG and QYL designed experiments. LNW performed experiments, analyzed results and wrote the manuscript. BC, HSD and XPC participated in experimental design and statistical collection. All authors read and approved the final manuscript.

Author information

Authors and Affiliations

Authors

Corresponding authors

Correspondence to Qingyun Li, Sansheng Geng or Xiaofen Zhang.

Ethics declarations

Conflict of interest

All authors report no declaration of interest.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Wang, L., Chen, B., Du, H. et al. Chromosome doubling of pepper haploids via chemical or high-temperature treatment. Euphytica 217, 146 (2021). https://doi.org/10.1007/s10681-021-02864-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s10681-021-02864-2

Keywords

Navigation