Production of doubled haplois in Brassica purpuraria

Abstract

Relevance. In recent years vegetable crop Brassica rapa ssp. chinensis var. purpuraria (synonyms: Brassica campestris L. var.purpurea Bailey; Brassica rapa L. ssp. chinensis var. purpurea) is gaining popularity as an object of genetic and molecular researches, and as an economically valuable vegetable plant due to the high content of biologically active compounds and distinctive economically valuable traits. Effective technology for development DH-plants to accelerate the breeding process for this culture has not been developed yet, so research in this area is relevant.Materials and methods. The study included two varieties from the collection of Vavilov AllRussian Research Institute of Plant Industry (VIR): No. 1301 (China) and No. 1357(Netherlands). Both protocols standard unmodified and with addition of silver nitrate (AgNO3) in the medium for embryogenesis induction were used in experiments for production of DH plants from isolated microspore in vitro. Direct chromosome counting in meristem cells and flow cytometry were used to determine the ploidy of regenerating plants.Results. As a result of the study embryogenesis in B. purpuraria culture can develop with the use of a standard protocol as well as with the addition of silver nitrate that showed a positive effect on the induction of embryogenesis. The yield of embryoids varied depending on the genotype of the individual plant within the variety accession. The highest yield of embryoids was 40 embryoids/petri dish. The main problem at the stage of regeneration is that about half of the regenerating plants occurred to be albinos and were not viable. We show a high degree of spontaneous chromosome doubling in regenerated plants (all analyzed plants were doubled haploids). In total 38 regenerated plants were obtained from accession No. 1301. It was shown that four DH-plants had self-incompatibility after self-pollination, but seed progeny from other plants was obtained. The created material was taken for genetics study and breeding work.

Haploid Embryogenesis in Isolated Microspore Culture of Carrots (Daucus carota L.)

“…The appearance of albino plants in dicotyledonous plants formed from microspores is a rather rare phenomenon. We are aware of the formation of albino seedlings obtained in an isolated culture of Brassica purpuraria microspores [ 58 ]. However, in our experiments, there were 63% albinos in Nantskaya 4 carrots.…”

Androgenesis of Red Cabbage in Isolated Microspore Culture In Vitro

“…The basic technology protocol for producing doubled haploids using isolated microspore culture in vitro, developed in the early 1980s [ 7 ] for rapeseed, was subsequently adapted by scientists for different members of the genus Brassica L. [ 8 , 9 , 10 ]. In the Russian Federation, this technique has been successfully applied to members of the Brassicaceae family, and F1 hybrids based on doubled haploid cabbage lines of white cabbage, Chinese cabbage, broccoli, and kohlrabi were obtained [ 11 , 12 , 13 , 14 , 15 , 16 , 17 , 18 , 19 , 20 ]. As early studies show, the protocol is not standard and needs to be adapted to the specific species and genotype.…”

The patriarch of Russian vegetable growing has a centenary

“…В 90-е годы проведены первые исследования по получению DH-растений овощных культур [19,20], на основе которых в дальнейшем были получены удвоенные гаплоидные растения сортообразцов моркови различного происхождения -НИИОХ 336, Витаминная, Московская зимняя А-515, Лосиноостровская 13, Леандр, Шантанэ 2461, Напе, Рондо, гибриды F1 Каратан, Калисто и другие [21]. В настоящее время разработаны технологии получения удвоенных гаплоидов через культуру микроспор in vitro для семейства капустные [22], включая капусту белокочанную [23,24], брокколи [25], репу [26], капусту пурпурную [27], горчицу сарептскую, индау посевной [28], редис европейский [29,30]. Разработан базовый протокол культуры микроспор in vitro для сортов салата (Lactuca sativa L.) [31].…”