Development of embryoids by microspore and anther cultures of red beet (Beta vulgaris L. subsp. vulgaris)

Górecka, Krystyna; Krzyżanowska, D.; Kowalska, Urszula; Kiszczak, Waldemar; Podwyszyńska, Małgorzata

doi:10.5513/jcea01/18.1.1877

Cited by 6 publications

(6 citation statements)

References 10 publications

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“…Sugar beet doubled haploid production through androgenesis has been attempted many times (Aflaki et al, 2017). Although all the androgenic attempts failed, recently androgenesis from sugar beet was tried by Górecka et al (2017). In spite of inducing dozens of androgenic embryoids and calli, none of them regenerated or even survived.…”

Section: The Effect Of Proline On Shoot Proliferationmentioning

confidence: 99%

“…In spite of inducing dozens of androgenic embryoids and calli, none of them regenerated or even survived. A genotypic effect on failure was not refuted and the inefficiency of the protocol was not denied (Górecka et al, 2017). The treated explants of haploid and doubled haploid can be decreased by necrosis (Klimek-Chodacka and Baranski, 2013).…”

Section: The Effect Of Proline On Shoot Proliferationmentioning

confidence: 99%

“…In sugar beet micropropagation, CKs are critical for inducing proliferation and its follow-up propagation (Gürel et al, 2008). However, the concentration of CKs needs to be finetuned for this species; otherwise, applying the hormone at higher concentrations can result in callogenesis, hyperhydricity, difficult rhizogenesis, necrosis, abnormal growth, and inefficient acclimation (Pospíšilová et al, 2000;Klimek-Chodacka and Baranski, 2013;Górecka et al, 2017;Pazuki et al, 2018a). Therefore, to maximize efficiency, CKs should be applied at an optimum level to increase proliferation/propagation while minimizing the side effects.…”

Section: Introductionmentioning

confidence: 99%

“…at higher concentrations can result in callogenesis, hyperhydricity, difficult rhizogenesis, necrosis, abnormal growth, and inefficient acclimation (Pospíšilová et al, 2000;Klimek-Chodacka and Baranski, 2013;Górecka et al, 2017;Pazuki et al, 2018a). Therefore, to maximize efficiency, CKs should be applied at an optimum level to increase proliferation/propagation while minimizing the side effects.…”

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confidence: 99%

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The effects of proline on in vitro proliferation and propagation of doubled haploid sugar beet (Beta vulgaris)

2018

Turk J Bot

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Doubled haploid induction is one of the available methods normally used for sugar beet breeding. Gynogenic haploid explants of sugar beet induced with 1 or 2 mg L -1 6-benzylaminopurine were treated with 5 g L -1 colchicine, then subcultured on a solidified MS medium plus 0.2 mg L -1 kinetin. Colchicine doubled the chromosome number of 27.7% of the treated haploid explants. With the aim of increasing the number of doubled haploid explants, the effects of five levels of proline (0.0, 0.1, 0.2, 0.3, or 0.4 mM) on the explants' proliferation, propagation, and shoot length were compared. With a large effect size (ES), proline at 0.3 mM induced the highest amount of proliferation, while proline-free medium resulted in the lowest amount of it. The highest propagation rates were observed for the explants treated on media with 0.2 and 0.3 mM proline (very large ES). Proline at 0.3 mM induced the shortest shoots (medium ES). A very strong positive correlation between proliferation and propagation, a moderate negative correlation between proliferation and length, and a strong negative correlation between propagation and length were observed. For the first time our results show beneficial effects of proline on in vitro proliferation and propagation of sugar beet.

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Section: The Effect Of Proline On Shoot Proliferationmentioning

confidence: 99%

Section: The Effect Of Proline On Shoot Proliferationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

mentioning

confidence: 99%

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The effects of proline on in vitro proliferation and propagation of doubled haploid sugar beet (Beta vulgaris)

2018

Turk J Bot

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“…At present, the economic importance of red beet requires the introduction of modern biotechnological methods into the breeding process with the possibility of obtaining doubled haploid plants for accelerated synthesis of high-yield hybrids. There is limited, sporadic literature data regarding the application of the gynogenesis and androgenesis method in red beet [48,49]. In addition, these publications provide only data on the induction of the gynogenesis process in in vitro unpollinated ovule culture and androgenesis in microspore culture; however, no information exists regarding obtained DH plants of red beet.…”

Section: Introductionmentioning

confidence: 99%

Production of Gynogenic Plants of Red Beet (Beta vulgaris L.) in Unpollinated Ovule Culture In Vitro

Zayachkovskaya

Домблидес

Zayachkovsky

et al. 2021

Plants

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The unique and balanced components of the biochemical composition, together with high antioxidant activity, make the red beet necessary a dietary vegetable crop, much contributing to healthy food ration. The application of the technology for producing gynogenic plants in vitro increases the genetic diversity and significantly reduces the period of time required to obtain the appropriate homozygous lines used to create the F1 hybrids that are demanded in the market. For induction of gynogenesis, we used IMB medium developed by us with the addition of 55 g/L sucrose, 3 g/L phytogel, 200 mg/L ampicillin, and 0.4 mg/L thidiazuron (TDZ) and cultured at 28 °C in the dark for 4–6 weeks. Shoot regeneration from embryoids and callus was performed on MS medium with 20 g/L sucrose, 3 g/L phytogel, 1 mg/L 6-benzylaminopurine (BAP), and 0.1 mg/L gibberellic acid (GA3). Immersion of the obtained microshoots with 5–7 well-developed leaves for 10–15 s into concentrated sterile indole-3-butyric acid (IBA) solution (50 mg/L) followed by their cultivation on solid medium ½ IMB with 2% sucrose and 3 g/L phytogel was the most efficient method for root formation. The addition of silver nitrate (22 mg/L) to the nutrient medium provoked an increase in the number of induced ovules up to nine per Petri dish (up to 25% of induced ovules). Gynogenic development was produced in six out of 11 genotypes studied, and the plants that were then acclimatized to ex vitro conditions were obtained in three genotypes (Nezhnost’, Dobrynya, b/a 128). The evaluation of ploidy of gynogenic plants that was carried out by flow cytometry and direct counting of chromosomes stained with propion-lacmoide revealed that all obtained gynogenic plants were haploids (2n = x = 9).

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Towards isolated microspores culture of red beet

2022

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Производство удвоенных гаплоидов (УГ) in vitro – ускоренный способ создания чистых линий для селекции коммерческих F1гибридов. Наиболее распространенный метод производства удвоенных гаплоидов у растений рода Beta, главным образом свеклы сахарной, – культура изолированных семязачатков (гиногенез). Недостатки гиногенного способа производства УГ: высокая трудоемкость вследствие ручной изоляции и инокуляции семязачатков на питательную среду и высокая вероятность развития соматических клонов из тканей, окружающих зародышевый мешок семязачатка. Значимая альтернатива для исключения описанных недостатков – культура изолированных микроспор (КИМ). Однако на настоящий момент протоколов рутинного производства УГ свеклы столовой в культуре изолированных микроспор не разработано. Цель исследования: изучение факторов, влияющих на каллусогенез в культуре изолированных микроспор свеклы столовой, и выявление оптимальных условий культивирования микроспор in vitro. В работе нами изучена связь размера бутона, стадии развития микроспор с частотой каллусогенеза методом микроскопирования и культивирования в питательной среде микроспор, выделенных из бутонов различной длины в пределах 1,2–2,7 мм с шагом в 0,3 мм. Культивированием микроспор на питательной среде NLN с добавлением 2,4-Д 0,1 мг/л, НУК 0,1 мг/л, 130 г/л сахарозы и на среде NLN без добавления регуляторов роста, содержащей 130 г/л сахарозы, исследовали влияние питательной среды, температуры культивирования и экспозиции температурной обработки на частоту каллусогенеза изолированных микроспор свеклы столовой. В результате исследования установлены размеры цветковых бутонов 1,2–1,5 мм, содержащие наиболее отзывчивые микроспоры одноядерной стадии развития, и питательная среда NLN с добавлением 130 г/л сахарозы, 0,1 мг/л 2,4-Д и 0,1 мг/л НУК, обеспечивающая наибольший выход каллуса в культуре изолированных микроспор свеклы столовой. Показано, что при примененных условиях температурная обработка изолированных микроспор при разной экспозиции при 32,5 °C не индуцирует эмбриогенез микроспор. Doubled haploids (DH) production in vitro is an accelerated way to create pure lines for breeding commercial F1 hybrids. The most common method for the production of doubled haploids in plants of the genus Beta, mainly sugar beet, is the culture of isolated ovules (gynogenesis). The disadvantages of the gynogenic method for the DH production are high labor intensity due to manual isolation and inoculation of ovules on a nutrient medium and a high probability of development of somatic clones from the tissues surrounding the embryo sac. To eliminate the described disadvantages, the culture of isolated microspores is a significant alternative. However, protocols for the routine DH production of red beet in culture of isolated microspores have not yet been developed. The aim of this study was to study the factors affecting callusogenesis in the culture of isolated red beet microspores and to identify the optimal conditions for their cultivation in vitro. Microspores were isolated from buds 1,2–2,7 mm with a step of 0,3 mm to study the relationship between the bud size and the stage of microspores development with the frequency of callusogenesis and 1,2–1,5 mm to study of the nutrient medium effect, cultivation temperature and the effect of exposure to heat treatment at 32,5 °C on callusogenesis of isolated red beet microspores. Microspores were cultivated on NLN nutrient medium supplemented with 2,4-D 0,1 mg/l, NAA 0,1 mg/l, 130 g/l sucrose and on NLN medium without the growth regulators addition containing 130 g/l sucrose. As a result of the study, the sizes of flower buds were found to be 1,2–1,5 mm, containing the most responsive microspores of the mononuclear stage of development to IMT, and the nutrient medium NLN with the addition of 130 g/l of sucrose, 0,1 mg/l of 2,4-D and 0,1 mg/l NAA, providing the highest yield of microsporogenic callus. It has been shown that under the applied conditions, thermal treatment of isolated microspores at different exposures at 32,5 °C does not induce microspore embryogenesis.

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Development of embryoids by microspore and anther cultures of red beet (Beta vulgaris L. subsp. vulgaris)

Cited by 6 publications

References 10 publications

The effects of proline on in vitro proliferation and propagation of doubled haploid sugar beet (Beta vulgaris)

The effects of proline on in vitro proliferation and propagation of doubled haploid sugar beet (Beta vulgaris)

Production of Gynogenic Plants of Red Beet (Beta vulgaris L.) in Unpollinated Ovule Culture In Vitro

Towards isolated microspores culture of red beet

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