Protocol of European Radish (Raphanus sativus L.) Microspore Culture for Doubled Haploid Plant Production

Kozar, E. V.; Домблидес, Е. А.

doi:10.1007/978-1-0716-1335-1_13

Cited by 4 publications

(3 citation statements)

References 8 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The beakers are placed on the magnetic stirrer for a short time. It is necessary for the magnet to break the integrity of the buds so that the microspores can enter the suspension [13].…”

Section: Microspore Isolation Stagementioning

confidence: 99%

“…The uneven maturation of microspores in the anthers of buds is expressed in different degrees in different cultures, and for such cultures where the unevenness of microspore development is significant, it can play a decisive role in the success of IMC technology [11]. It also leads to a narrowing of the suitable bud size range, which technically complicates the technology [12,13]. There are a number of studies that have attempted to separate only developmental stages responsive to embryogenesis from microspore suspension, but these modifications are difficult to apply in practice and require expensive reagents, limiting their use in extensive practice [14][15][16].…”

Section: Introductionmentioning

confidence: 99%

“…This inevitably occurs due to mechanical effects on bud tissues during isolation of microspores. In standard protocols for obtaining doubled haploids through in vitro microspore culture, isolation of microspores occurs by breaking buds immersed in nutrient medium: in magnetic stirrer boxes [13]; by compressing buds with a rotary motion in test tubes using a piston (syringe) [17]; by grinding buds with a pestle in a mortar [18]; by crushing buds with a glass rod in a test tube [19]; or by crushing buds in a blender [20].…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Effect of the Method of Microspore Isolation on the Efficiency of Isolated Microspore Culture In Vitro for Brassicaceae Family

Kozar

Домблидес

2022

Horticulturae

View full text Add to dashboard Cite

Isolated microspore culture in vitro (IMC) is an advanced technique for producing doubled haploids. We developed a modified microspore isolation method for the Brassicaceae family, which exceeds the results obtained by a standard microspore isolation method. We found that the new method allows an increase in the percentage of microspores at the embryogenic stage of development in the culture. In the spring rapeseed ‘Ratnik’ culture the percentage of microspores increases from 66.7% to 73%, and in the European radish ‘RBK’ from 34% to 61.9%. Moreover, the new method of microspore isolation made it possible to expand the range of linear bud sizes (from 3.5–4.0 to 3.0–4.5 mm for spring rapeseed ‘Ratnik’) suitable for IMC technology. In addition, the new method of microspore isolation reduced the debris in the preparation of spring rapeseed ‘Ratnik’ and European radish ‘RBK’ by 2.4 and 15 times, respectively. The best results were shown on Sareptian mustard No. 72, where the yield of embryoids increased by 7.5 times. Remarkably, the new method of microspore isolation allowed us to obtain the first embryoids of red cabbage No. 428, whereas no embryoids were obtained using the standard method of microspore isolation. In summary, the new method of microspore isolation allows an increase in the efficiency of IMC technology for Brassicaceae family crops.

show abstract

“…The beakers are placed on the magnetic stirrer for a short time. It is necessary for the magnet to break the integrity of the buds so that the microspores can enter the suspension [13].…”

Section: Microspore Isolation Stagementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Effect of the Method of Microspore Isolation on the Efficiency of Isolated Microspore Culture In Vitro for Brassicaceae Family

Kozar

Домблидес

2022

Horticulturae

View full text Add to dashboard Cite

show abstract

Modern trends in the development of selection of vegetable and melon crops

et al. 2022

View full text Add to dashboard Cite

The article reflects the issues of the current state of scientific research in the FSBSI FSVC, directions and prospects for the development of selection and seed production of vegetable crops are shown. Varieties have been created that realize the bioclimatic potential of the cultivation zone, meet market requirements, and are capable of producing high yields under the influence of biotic and abiotic stressors. Selection is carried out in a combination of classical methods with innovative technologies.Biotechnologists have improved the basic protocol for in vitro microspore culture for the Brassicaceae, stages of technology for obtaining doubled haploids in microspore culture in vitro for the Apiaceae are being developed. For the first time, an effective protocol for obtaining double haploids of European radish has been developed. Doubled haploids of vegetable crops of the Cucurbitaceae, Amarantháceae, Amaryllidáceae were obtained in vitro in the culture of unpollinated ovules. The Center conducts research on the increased content of biologically active substances and antioxidants. An important place is occupied by immunological studies on the resistance of vegetable crops to harmful diseases. The institution develops systems of protective measures against pests, diseases and weeds for vegetable crops based on the use of new environmentally friendly preparations. On the basis of fundamental and priority applied research of previous years, the creation of 21 varieties and hybrids of cabbage, table beet, tomato, sweet pepper, onion, chives, leek, cucumber, melon, lettuce, chervil, potato, aster, lychnis has been completed. Along with the creation of new breeding achievements, technologies for their cultivation are being improved through the use of new agricultural practices, microfertilizers, biological products and humates on vegetable crops in various soil and climatic zones of Russia. New agricultural methods are being developed in the cultivation of mushrooms. In the field of seed production, methods are being developed to increase the germination of seeds, seed production, elements of the technology for the production of mother liquors and seeds of various vegetable and melon crops; zonal seed production technologies that increase their yield. In 2021, a team of scientists received an award from the Government of the Russian Federation for scientific work "Development and implementation of innovative technologies for growing vegetables and potatoes to provide the population with environmentally friendly food".

show abstract

Thidiazuron Promoted Microspore Embryogenesis and Plant Regeneration in Curly Kale (Brassica oleracea L. convar. acephala var. sabellica)

et al. 2023

View full text Add to dashboard Cite

Curly kale (Brassica oleracea L. convar. acephala var. sabellica), the most common type of edible kale, characterized by providing rich nutrition and health care functions, is sought after and has been listed as top of the healthiest vegetables in recent trends, and has aroused the interest of breeders in cultivating new varieties. However, it usually takes more than six years to obtain a homozygous kale inbred line for commercial seed production through conventional breeding procedures due to its long growth and development period. The isolated microspore culture (IMC) technique could be a time-saving alternative method for producing doubled haploid (DH) lines that are genetically homozygous. In this study, we successfully utilize the efficient cytokinin thidiazuron (TDZ) to promote microspore embryogenesis and plant regeneration in two curly kale cultivars (‘Winterbor F2’ and ‘Starbor F2’). Compared with the control (0 mg/L TDZ), all tested TDZ concentrations (0.1, 0.2, 0.3, 0.4 mg/L) had no adverse effects on embryogenesis, and 0.2 mg/L TDZ had an optimal effect on embryo survival and plant regeneration of the two genotypes. For ‘Starbor F2’, 0.2 mg/L TDZ treatment achieved the highest embryogenesis rate (1.83-fold higher than the control group) and direct seeding rate (1.61-fold increase), and the lowest mortality rate. Likewise, 0.2 mg/L TDZ increased the embryogenesis rate of ‘Winterbor F2’ by 1.62 times, the direct seeding rate by 1.61 times, and the mortality rate fell to the lowest. A 1/2 Murashige and Skoog (MS) medium with 0.2 mg/L 1-Naphthaleneacetic acid (NAA) can significantly promote the rooting of the regenerated seedlings. These results provide new insights into the practical application of the IMC technique in shortening the breeding cycle of kale.

show abstract

Protocol of European Radish (Raphanus sativus L.) Microspore Culture for Doubled Haploid Plant Production

Cited by 4 publications

References 8 publications

Effect of the Method of Microspore Isolation on the Efficiency of Isolated Microspore Culture In Vitro for Brassicaceae Family

Effect of the Method of Microspore Isolation on the Efficiency of Isolated Microspore Culture In Vitro for Brassicaceae Family

Modern trends in the development of selection of vegetable and melon crops

Thidiazuron Promoted Microspore Embryogenesis and Plant Regeneration in Curly Kale (Brassica oleracea L. convar. acephala var. sabellica)

Contact Info

Product

Resources

About