Microspore Culture and Doubled Haploid Technology

Takahata, Yoshihito; Takahashi, Yu; Tsuwamoto, Ryo

doi:10.1007/978-1-4614-7795-2_4

Cited by 3 publications

(3 citation statements)

References 76 publications

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“…The most basic requirement for the application of the gene gun-mediated method to the genetic transformation of Chinese cabbage is that there is no significant effect on the vigor of new explants after the bombardment of the gene gun, which can induce the production of regenerated plants. There are a series of factors that influence microspores cultures, and we established a mature microspore culture system based on our former study [ 44 , 49 ]. In this experiment, the viability of microspores and the ability of microspores to induce embryo regeneration after gene gun bombardment were identified.…”

Section: Discussionmentioning

confidence: 99%

Efficient transformation of the isolated microspores of Chinese cabbage (Brassica rapa L. ssp. pekinensis) by particle bombardment

Liu,

Zhang,

Zhang

et al. 2024

Plant Methods

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Background The low efficiency of genetic transformation in Chinese cabbage (Brassica rapa L. ssp. pekinensis) is the key problem affecting functional verification. Particle bombardment is a widely used method along with the Agrobacterium-mediated method. As a physical means, it has almost no restrictions on the type of host and a wide range of receptor types, which largely avoids the restriction of explants. The bombardment parameters, which include the number of bombardments, the bombardment pressure, and the bombardment distance, may affect the microspores' genetic transformation efficiency. Results The transformation efficiency was improved using the particle bombardment method under the combination of bombardment shot times (3, 4, 5) × bombardment pressure (900, 1100, 1350 psi) × bombardment distance (3, 6, 9 cm). The average viability of microspores in the treatment group ranged from 74.76 to 88.55%, while the control group was 88.09%. When the number of shot times was 4, the number of embryos incubated in the treatment group ranged from 16 to 236 per dish, and the control group had 117 embryos per dish. When the bombardment parameters of the biolistic method were 4 shot times—1350 psi—3 cm, 4 times—1100 psi—3 cm, and 4 times—900 psi—3 cm, they had high transient expression efficiency, and the average number of transformed microspores was 21.67, 11.67, and 11.67 per dish (3.5 mL), respectively. When the bombardment parameters were 4 times, 900 psi, and 6 cm, the highest genetically transformed embryos were obtained, and the transformation efficiency reached 10.82%. Conclusion A new genetic transformation system with proper parameters for Chinese cabbage microspores was established using particle bombardment. This proper transformation system could provide a useful tool for the improvement of cultivar quality and the investigation of functional genes in Chinese cabbage.

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Section: Discussionmentioning

confidence: 99%

Efficient transformation of the isolated microspores of Chinese cabbage (Brassica rapa L. ssp. pekinensis) by particle bombardment

Liu,

Zhang,

Zhang

et al. 2024

Plant Methods

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“…It was concluded that BBM genes activate developmental pathways leading to plant growth; therefore, they may be the starting point of microspore redifferentiation [71]. The LEAFY COTYLEDON genes (LEC1 and LEC2), EMBRYOMAKER (EMK), FUSCA3, and SHOOTMERISTEMLESS (STM) seem to be related to microspore-derived embryo development; however, they have not been correlated with the induction of microspore embryogenesis [29,[72][73][74][75][76]. Other genes discovered through analysis of differentially expressed genes (DEGs) have been reported to carry unknown function and have never been connected with microspore embryogenesis [77].…”

Section: Gene Expression Changesmentioning

confidence: 99%

Brassica napus Haploid and Double Haploid Production and Its Latest Applications

Starosta,

Szwarc,

Niemann

et al. 2023

CIMB

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Rapeseed is one of the most important oil crops in the world. Increasing demand for oil and limited agronomic capabilities of present-day rapeseed result in the need for rapid development of new, superior cultivars. Double haploid (DH) technology is a fast and convenient approach in plant breeding as well as genetic research. Brassica napus is considered a model species for DH production based on microspore embryogenesis; however, the molecular mechanisms underlying microspore reprogramming are still vague. It is known that morphological changes are accompanied by gene and protein expression patterns, alongside carbohydrate and lipid metabolism. Novel, more efficient methods for DH rapeseed production have been reported. This review covers new findings and advances in Brassica napus DH production as well as the latest reports related to agronomically important traits in molecular studies employing the double haploid rapeseed lines.

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“…It is shown that there is a high correlation between the number of chloroplasts in the stomata guard cells and the ploidy of plants [24]. Thus, the optimization of DHapproach for breeding and seed production remains relevant [25][26][27][28].…”

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confidence: 99%

NEW GENERATION HYBRIDS OF WHITE CABBAGE (Brassica oleracea L. convar. capitata var. alba DC) BASED ON DOUBLED HAPLOIDS

Pivovarov¹,

Бондарева²,

Шмыкова³

et al. 2017

S-h. biol.

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A b s t r a c tPresently, cabbage breeding is mainly focused on F 1 hybrids necessitating constant parental lines to be obtained. Doubled haploid (DH) technology based on isolated microspore in vitro culture is widely used to produce pure lines of brassica crops. This method allows us to rapidly develop homozygous lines, in contrast to time-consuming traditional breeding for heterosis in cross-pollinating crops which takes 7 to 10 years for annuals and 14 to 20 years for biennial plants. One of the objectives of DH technology is to provide the all possible number of doubled haploid plants that allows more fully encompass the spectrum of genetic recombination, including the recessive locus. The aim of our study was to evaluate economically important traits in white cabbage (Brassica oleracea L. convar. capitata var. alba DC) constant doubled haploid lines of late ripening and to improve the technology for producing DH based F 1 hybrids. Eleven breeding lines of late ripening cabbage were used to obtain doubled haploid lines from microspore in vitro culture. Of the obtained lines, twelve doubled haploid genotypes were selected for further use based on evaluation of ploidy and combining ability. Seed progeny was reproduced by hybridization of regenerated plants in a climatic chamber (2014)(2015). We used the schemes of creating self-incompatible lines and two-line-based hybrids. In the field trials (Moscow region, 2014(Moscow region, -2015, the doubled haploids and their hybrid combinations were compared to the standard (Severyanka F 1 ) for the main valuable characteristics (i.e. the content of dry matter, nitrates, and vitamin C). The field resistance to Fusarium wilt, alternariosis, and pest damage were determined at cabbagehead technical maturity. The resistance to clubroot was assessed under artificial infection. There was a direct relationship of the average number of chromosomes to the number of chloroplasts in the stomata guard cells and the length of guard cells. The frequency of spontaneous doubling of the chromosomes numbers varied from 50.0 % to 87.5 % in different geotypes. A total of 11 to 73 % produced lines were high self-incompatible. Their geitonogamic pollination in the topcrosses resulted in 42 hybrid combinations. The model of F 1 hybrid most fully responding to consumer market demands was developed. Ten promising hybrid combinations which matched the model parameters in two-year field testing were recommended for variety testing. Hybrids were characterized by uniformity, high biochemical quality, the resistance to major diseases and pests and the yield of 104.60±8.27 t/ha. The dry matter content reached to 10.5 %, the sugar content was about 4.21-5.10 %, and ascorbic acid level ranged from 21.12 to 38.70 mg%. Both the highest level of ascorbic acid (92.0 mg%) and the smallest nitrate accumulation (33 mg/kg) were characteristic of one hybrid combination.Keywords: white cabbage, Brassica oleracea L., doubled haploid lines, heterosis F 1 hybrids, in vitro isolated microspore culture, self-incomp...

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Microspore Culture and Doubled Haploid Technology

Cited by 3 publications

References 76 publications

Efficient transformation of the isolated microspores of Chinese cabbage (Brassica rapa L. ssp. pekinensis) by particle bombardment

Efficient transformation of the isolated microspores of Chinese cabbage (Brassica rapa L. ssp. pekinensis) by particle bombardment

Brassica napus Haploid and Double Haploid Production and Its Latest Applications

NEW GENERATION HYBRIDS OF WHITE CABBAGE (Brassica oleracea L. convar. capitata var. alba DC) BASED ON DOUBLED HAPLOIDS

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