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Horticulturae, volume 8, issue 11, pages 1083

Efficient Methods for Evaluation on Ploidy Level of Cucurbita pepo L. Regenerant Plants Obtained in Unpollinated Ovule Culture In Vitro

Domblides Elena ¹

Ermolaev Alexey ¹

Belov Sergey ¹

Kan Lyudmila ¹

Skaptsov Mikhail ²

Domblides Arthur ¹

Hide authors affiliations Show authors affiliations: 2 affiliations

Federal State Budgetary Scientific Institution Federal Scientific Vegetable Center (FSBSI FSVC), VNIISSOK, 143072 Moscow, Russia |

South Siberian Botanical Garden, Altai State University, 656049 Barnaul, Russia |

Publication type: Journal Article

Publication date: 2022-11-16

Multidisciplinary Digital Publishing Institute (MDPI)

Journal: Horticulturae

Quartile SCImago

Quartile WOS

Impact factor: 3.1

ISSN: 23117524

DOI: 10.3390/horticulturae8111083

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Plant Science

Horticulture

Abstract

An important stage in doubled haploid (DH) production is to evaluate and to differentiate the ploidy level of regenerant plants at least two–three times during the technology. Therefore, rapid and reliable methods are necessary for particular species taken into the technology. In this study, Cucurbita pepo regenerants obtained through unpollinated ovule culture in vitro were evaluated including three different methods: direct chromosome counting in apical meristems, flow cytometry of the cell nucleus, and estimation of morphological parameters of the abaxial epidermis. Methods were optimized for each of three evaluations, and main criteria were determined for ploidy level differentiation. As a result, four ploidy levels, namely, 2n, 3n, 4n, and 8n, were defined among regenerant plants adapted to ex vitro conditions, while true haploids were only found among plants that remained in the in vitro culture. In total, 32.35%, 26.47%, 33.82%, 4.41%, and 2.94% of regenerant plants of courgette and patisson were diploid, triploid, tetraploid, octaploid, and aneuploid, respectively. According to results of flow cytometry of the cell nucleus, two cytotypes in diploid samples with DNA content of 2C = 1.07 ± 0.03 pg for courgette belonging to subsp. pepo and 2C = 0.95 ± 0.03 pg for patisson samples belonging to subsp. ovifera were revealed. The images of metaphase chromosomes of haploid, triploid, and tetraploid C. pepo specimens obtained using the propion–lacmoid chromosome staining method were presented for the first time. Parameters of abaxial epidermis in diploid samples of courgette and patisson grown in open-field and greenhouse conditions were described and compared. It was shown that the most robust parameter not depending on external factors was the number of chloroplasts in stomatal guard cells, which contained 9.41 to 11.31, 14.84 to 16.3, and up to 17.58 chloroplasts in diploid, triploid, and tetraploid samples, respectively. The application of several methods for estimation enables avoiding the misidentification of ploidy levels in adapted regenerant plants produced with the use of DH technology.

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Euphytica	Euphytica, 1, 33.33% Euphytica 1 publication, 33.33%
Vegetable crops of Russia	Vegetable crops of Russia, 1, 33.33% Vegetable crops of Russia 1 publication, 33.33%
Horticulturae	Horticulturae, 1, 33.33% Horticulturae 1 publication, 33.33%
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Springer Nature	Springer Nature, 1, 33.33% Springer Nature 1 publication, 33.33%
Federal Scientific Center for Vegetable Growing	Federal Scientific Center for Vegetable Growing, 1, 33.33% Federal Scientific Center for Vegetable Growing 1 publication, 33.33%
Multidisciplinary Digital Publishing Institute (MDPI)	Multidisciplinary Digital Publishing Institute (MDPI), 1, 33.33% Multidisciplinary Digital Publishing Institute (MDPI) 1 publication, 33.33%
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Domblides E. et al. Efficient Methods for Evaluation on Ploidy Level of Cucurbita pepo L. Regenerant Plants Obtained in Unpollinated Ovule Culture In Vitro // Horticulturae. 2022. Vol. 8. No. 11. p. 1083.

GOST all authors (up to 50) Copy

Domblides E., Ermolaev A., Belov S., Kan L., Skaptsov M., Domblides A. Efficient Methods for Evaluation on Ploidy Level of Cucurbita pepo L. Regenerant Plants Obtained in Unpollinated Ovule Culture In Vitro // Horticulturae. 2022. Vol. 8. No. 11. p. 1083.

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RIS Copy

TY - JOUR

DO - 10.3390/horticulturae8111083

UR - https://doi.org/10.3390%2Fhorticulturae8111083

TI - Efficient Methods for Evaluation on Ploidy Level of Cucurbita pepo L. Regenerant Plants Obtained in Unpollinated Ovule Culture In Vitro

T2 - Horticulturae

AU - Domblides, Elena

AU - Ermolaev, Alexey

AU - Kan, Lyudmila

AU - Skaptsov, Mikhail

AU - Domblides, Arthur

AU - Belov, Sergey

PY - 2022

DA - 2022/11/16 00:00:00

PB - Multidisciplinary Digital Publishing Institute (MDPI)

SP - 1083

IS - 11

VL - 8

SN - 2311-7524

ER -

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@article{2022_Domblides,

author = {Elena Domblides and Alexey Ermolaev and Lyudmila Kan and Mikhail Skaptsov and Arthur Domblides and Sergey Belov},

title = {Efficient Methods for Evaluation on Ploidy Level of Cucurbita pepo L. Regenerant Plants Obtained in Unpollinated Ovule Culture In Vitro},

journal = {Horticulturae},

year = {2022},

volume = {8},

publisher = {Multidisciplinary Digital Publishing Institute (MDPI)},

month = {nov},

url = {https://doi.org/10.3390%2Fhorticulturae8111083},

number = {11},

pages = {1083},

doi = {10.3390/horticulturae8111083}

}

MLA

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Domblides, Elena, et al. “Efficient Methods for Evaluation on Ploidy Level of Cucurbita pepo L. Regenerant Plants Obtained in Unpollinated Ovule Culture In Vitro.” Horticulturae, vol. 8, no. 11, Nov. 2022, p. 1083. https://doi.org/10.3390%2Fhorticulturae8111083.

Found error?

Publisher

Multidisciplinary Digital Publishing Institute (MDPI)

Journal

Horticulturae

Quartile SCImago

Quartile WOS

Impact factor

3.1

ISSN

23117524 (Electronic)

Labs

Laboratory of reproductive biotechnology in the breeding of agricultural plants

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