Assessment of different anther culture approaches to produce doubled haploids in cucumber (Cucumis sativus L.)

Asadi, Abouzar; Zebarjadi, Alireza; Abdollahi, M.R.; Seguí-Simarro, José M.

doi:10.1007/s10681-018-2297-x

Cited by 17 publications

(29 citation statements)

References 23 publications

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“…У культур семейства Cucurbitaceae также имеются сообщения об успешном использовании SSR-маркеров для оценки растений-регенерантов, полученных в культуре изолированных пыльников огурца [134] и неопыленных завязей/семяпочек дыни [116] и огурца Cucumis sativus L. [107].…”

Section: использование молекулярного анализа для оценки происхождения/гомозиготности растений полученных с использованием Dh-технологийunclassified

“…Микросателлитные праймеры оказались удобными для оценки растений, полученных не только в культуре изолированных неопыленных завязей/семяпочек, но и в культуре изолированных пыльников огурца. Из 15 протестированных SSR-праймеров два (CMAG59 и CMCTT144) оказались полиморфными и, следовательно, полезными для идентификации DH среди популяции растений [134]. Как и ожидали авторы, согласно оригинальному описанию Danin-Poleg et al [135], растения-доноры имели три полосы амплификации для CMAG59, тогда как растения, полученные из пыльников, представили одну/две (для DH) или три исходные полосы (для соматических регенерантов).…”

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Obtaining doubled haploids of Cucurbita pepo L.

2021

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Doubled haploids have been widely used worldwide in breeding programs and fundamental research as valuable homozygous material for about 100 years. The species Cucurbita pepo L. are represented by a huge variety of forms, include highly productive vegetable crops and have a wide distribution in the world. Despite the great economic importance, the creation of effective protocols to ensure stable production of doubled haploids in this species remains an urgent task. DH plants are of interest not only because of the acceleration of the breeding process, but also because of the realization of the huge potential of gametoclonal variability inherent in this highly polymorphic species. In this review, we analyzed the main technologies used for obtaining doubled haploids in vegetable crops of C. pepo: parthenogenesis in situ stimulated by treated/irradiated pollen, gynogenesis in vitro (unpollinated ovule culture in vitro) and androgenesis in vitro (anther/microspore culture in vitro). An analysis is presented of the research carried out from the beginning of the discovery of haploid plants to the current advances and evaluation of the prospects in the field of DH plant production. The main critical factors influencing the efficiency of each technology and its individual steps are considered. The developed technology of doubled haploids obtaining using non-pollinated ovary culture in vitro is presented. This technology allows to obtain up to 55 embryoids per one cultivated ovary (28 embryoids/ 100 cultivated ovules) To introduce haploid technologies into the breeding process it is necessary to evaluate the obtained plants for ploidy level. The use of direct counting of chromosomes in apical cells may present a certain difficulty in this species due to their large number (2n=40) and their small size. Depending on the level of laboratory equipment, ploidy determination using flow cytometry of cell nuclei and counting the number of chloroplasts in stomatal guard cells in the epidermis of the abaxial side of the leaf may be more convenient methods. The prospects for the use of molecular markers for assessment for homozygosity in DH technologies used, including C. pepo, are discussed in the review.

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Obtaining doubled haploids of Cucurbita pepo L.

2021

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“…Stress is considered as a kind of mutation on microspore embryos: unconstrained microspores form flower powder along the normal binding pathway (Touraev et al, 1997). Among various stress treatments, heat stress is widely used to initiate microspore embyogenesis (ME) in many crops (Asadi et al, 2018;Bhatia et al, 2018;Cimò et al, 2017;Dubas et al, 2014). The anthers of sweet pepper cultured under heat stress can be induced to develop into haploid embryos with complete functions instead of mature pollen (Bárány et al, 2005).…”

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confidence: 99%

Exploring differentially expressed genes of microspore embryogenesis under heat stress in sweet pepper

Yan¹,

Yan-sheng²,

Ru-yi³

et al. 2020

Afr. J. Biotechnol.

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Stress is considered to be the inducer of microspore embyogenesis (ME), and heat stress is indispensible in the ME of sweet pepper. The aim of the study was to explore differentially expressed genes of microspore embryogenesis under heat stress in sweet pepper. The swollen rate of microspore was significantly affected by heat stress, while no green plant could be acquired without heat pretreatment. Anthers with or without heat stress were used for whole transcriptome analysis by RNA sequencing to provide new insights on how cells adapt to stress. A total of 5031 differentially expressed genes were identified, among which 2657 differentially expressed genes were up-regulated and 2374 differentially expressed genes were down-regulated in the early stage of heat stress. KEGG pathway analysis identified "plant hormone signal transduction" (67; 11.20%), followed by starch and sucrose metabolism (63; 10.54%). RNA-Seq data and quantitative real-time polymerase chain reaction showed that 224 genes related to glutathione metabolism, starch and sucrose metabolism, plant hormone signal transduction and phenylpropanoid biosynthesis were the most likely specific genes in ME under heat stress. This research provides new insights into molecular regulation during the early stage of ME in sweet pepper under heat stress.

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“…Эмбриоиды и морфогенные каллусы получают в культуре пыльников или микроспор на питательных средах, содержащих различные концентрации 6-бензиламинопурина (БАП/BAP), 2,4-дихлорфеноксиуксусной кислоты (2,4-Д/2,4-D) и кинетина (КТ). За андрогенезом следует стадия дифференцировки, для которой не желательно присутствие в питательной среде 2,4-Д, поэтому в питательные среды обычно добавляют (различные варианты по концентрации и соотношению между собой) БАП, 1-нафталинуксусной кислоты (НУК/NAA) и KТ (Kumar & Murthy, 2004;Kumar et al al., 2003;Song et al, 2007;Abdollahi et al, 2016;Asadi et al, 2018).…”

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“…Kumar et al, 2003; AshokKumar and Murthy, 2004;Song et al, 2007;Zhan et al, 2009;Rakha et al, 2012;Hamidvand et al, 2013; Abdollahi et al, 2015Abdollahi et al, , 2016Kurtar et al, 2016;Asadi et al, 2018;Amirian et al, 2019). Однако выход растений-регенерантов остается достаточно низким и очень сильно зависит от генотипа донорного растения(Abdollahi et al, 2016;Kumar & Murthy, 2004;Kumar et al, 2003;Song et al, 2007) Asadi et al, 2018…”

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Production of Doubled Haploids in cucumber

et al. 2019

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Implementation of cell technologies has essentially improved the plant breeding process in agricultural crops in the world. The production of pure lines in cultivated crops, particularly among cross-pollinated species such as cucumber (Cucumis sativus L.) requires much time, labor and expense. Thus, the use of DH-plants for production of fully homozygous lines for one year becomes a very promising method for near cucumber breeding program. The major factor limiting the wide use of DH is a lack of effective protocol for large-scale plant production. In this review the historical facts with description of three main methods of DH-plant production were presented. By now these three methods have been such as parthenogenesis in situ induced by pollination with irradiated or chemically treated pollen; androgenesis in vitro including anther and isolated microspore cultivation in vitro; gynogenesis through ovule cultivation in vitro. Comparative analysis of published data with regard to the efficiency of the technology for DH-plant production was shown as well as advantages and limitations of each technology were described.

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Assessment of different anther culture approaches to produce doubled haploids in cucumber (Cucumis sativus L.)

Abstract: AA performed all the experimental work and analyzed the results. AZ, MRA and JMSS designed the experimental work and analyzed the results. JMSS wrote the manuscript.

Cited by 17 publications

References 23 publications

Obtaining doubled haploids of Cucurbita pepo L.

Obtaining doubled haploids of Cucurbita pepo L.

Exploring differentially expressed genes of microspore embryogenesis under heat stress in sweet pepper

Production of Doubled Haploids in cucumber

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