Production of haploid embryos and plants in Iranian melon (Cucumis melo L.) through irradiated pollen-induced parthenogenesis.

2021 ◽  
pp. 127-133
Author(s):  
Leila Bagheri ◽  
Mahmoud Lotfi ◽  
Mansour Nori
Keyword(s):  
Liquid Culture ◽  
Gamma Ray ◽  
Culture Liquid ◽  
Plant Production ◽  
Haploid Plant ◽  
Culture Methods ◽  
Irradiated Pollen ◽  
Breeding Programmes ◽  
Haploid Embryos ◽  
Haploid Plants

Abstract The irradiated pollen technique (IPT) is the most successful haploidization technique within Cucurbitaceae. The influence of gamma-ray doses (250, 350, 450 and 550 Gy), genotypes and stage of development of embryos obtained by IPT on the induction of haploid embryos were studied in several Iranian melon cultivars as well as their hybrids with alien cultivars. Female flowers were pollinated using pollen that had been irradiated with gamma rays. Different shapes and stages of embryos were excised 21-25 days after pollination and cultured on E20A medium. Direct culture, liquid culture and integrated culture methods were used; integrated culture and liquid culture methods showed advantages in increasing the efficiency of haploid plant production in melon breeding programmes. Results revealed that 550 Gy of gamma irradiation was successful in inducing parthenogenesis and fruit development, whereas lower irradiation doses were not effective in inducing haploid embryos. The percentages of embryos per seed were the highest in 'Samsoori' (1.2%) and 'Saveh' (1.1%) cultivars. Some of the heart-shaped and cotyledon-shaped embryos developed into haploid plants. In total, 52 parthenogenic melon plantlets were recovered from 274 embryos via IPT. Production of haploid embryos and haploid plants was strongly influenced by gamma-ray dose, embryo stage and genotype. Indirect methods and chromosome counting performed on the root cells of regenerated plants showed that these plants were haploid (n = x = 12).

scholarly journals Factors Affecting Doubled Haploid Plant Production Via Maize Technique in Bread Wheat

2015 ◽  
Vol 56 (2) ◽  
pp. 67-73
Author(s):  
Ioannis Xynias ◽  
Antonios Koufalis ◽  
Evdokia Gouli-Vavdinoudi ◽  
Demetrios Roupakias
Keyword(s):  
Bread Wheat ◽  
Doubled Haploid ◽  
Embryo Rescue ◽  
Plant Production ◽  
Haploid Plant ◽  
In Planta ◽  
Maize Pollen ◽  
Doubled Haploid Plants ◽  
Haploid Embryos ◽  
Haploid Plants

Abstract The effect of two in planta factors (growth conditions, genotype) and two in vitro factors (time of embryo rescue, embryo rescue medium) on doubled haploid (DH) plant production in bread wheat via maize technique was investigated in nine F1 hybrids produced after crossing four bread wheat cultivars. During the first year one group of F1 plants was grown in a field and at the proper stage pollinated with maize pollen (sweet corn popu-lation). In parallel, a second group of F1 plants was grown in a growth chamber and pollinated as in the former group. In the second growing season the experiment was repeated but only field-grown plants were used. All the produced haploid embryos were cultured in three different media and the resulting 146 haploid plants were sub-sequently treated with aqueous solution of colchicine. Finally, 86 doubled haploid plants were obtained. We noted that the growing conditions of the parental plants and the intervening time between day of pollination and day of embryo rescue influenced the percentage of haploid embryo production. Culture medium also influenced haploid and doubled haploid plant production. The two media (MS/2, B5) were found equally effective. Most of the haploid embryos originated from the Penios × Acheloos cross, whereas most of the doubled haploid plants were produced from the KVZ × Penios cross. Doubled haploid plants were produced from all crosses.

scholarly journals Study of application times gibberellic acid and 2,4-Dichlorophenoxyacetic acid in the plant regeneration from wheat haploid embryos in chromosome elimination method

2020 ◽  
Author(s):  
Hamed Modirrousta ◽  
Raheleh Khademian ◽  
Reza Bozorgipour
Keyword(s):  
Gibberellic Acid ◽  
Chromosome Elimination ◽  
Plant Production ◽  
Haploid Plant ◽  
Dichlorophenoxyacetic Acid ◽  
Elimination Method ◽  
Significant Difference ◽  
Haploid Embryos ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Haploid Plants

AbstractWheat is one of the most important cereals, which is very valuable in food. Haploid plants are of particular importance in plant breeding. The wheat seeds produced in the crosses between wheat and maize in the chromosome elimination method without of endosperm and are immature embryo, to prevent the abortions haploid embryos, they must have embryo rescue. Increasing production of haploid plants from produced embryos can improve production efficiency. In this study, With attention their effects gibberellic acid and 2,4-Dichlorophenoxyacetic acid on growth, cell size and cell division, Their use in the production of wheat haploid plant were studied. There was a significant difference at level 1% between the not use and use of gibberellic acid in difference times in the production of haploid from embryos, So that the most haploid plant produced in the use of gibberellic acid in the 4 days after pollination. Also, the use of 2,4-Dichlorophenoxyacetic acid in tiller maintenance liquid culture medium was evaluated at times 48 and 72 hours after pollination. There were a significant difference between these treatments at the 1% level and the most was obtained for wheat haploid plant production with application of 2,4-Dichlorophenoxyacetic acid treatment for 72 hours.Highlightproduction of haploid plants plays an important role in wheat breeding. This technique is done to get doubled haploid and absolute homozygous plants in a very short duration of time.

scholarly journals First Report of Obtaining Haploid Plants Using Tissue Culture Techniques in Spinach

HortScience ◽  
2016 ◽  
Vol 51 (6) ◽  
pp. 742-749 ◽  
Author(s):  
Davut Keleş ◽  
Ceren Özcan ◽  
Hasan Pınar ◽  
Atilla Ata ◽  
Nihal Denli ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Anther Culture ◽  
Gamma Ray ◽  
Chromosome Doubling ◽  
Culture Methods ◽  
Hermaphrodite Flower ◽  
Culture Techniques ◽  
Irradiated Pollen ◽  
Tissue Culture Techniques ◽  
Haploid Plants

Monoic, dioic, and hermaphrodite flower types complicate spinach breeding and cultivar development. The availability of haploid plants will definitely accelerate spinach breeding; however, there are currently no reports in the literature about the use of tissue culture techniques to obtain spinach haploids. Therefore, in this study, pollination with irradiated pollen and anther culture methods were used to obtain haploid plants in spinach. For anther culture, three spinach varieties (Koto F1, Favorit F1, and Greenstar F1) and four different nutrient media were tested to obtain haploid embryos. Significant outcomes were not achieved from anther culture, and only two plants were obtained from all the experiments. Gynogenesis studies using irradiated pollen were performed with the same three spinach varieties and six gamma ray doses (100, 150, 200, 300, 400, and 500 Gy) from (cobalt60) Co60. Murashige and Skoog (MS) nutrient medium containing 1 mg·L−1 indoleacetic acid (IAA) was used for embryo germination. Current findings revealed that the varieties produced responded differently to the various doses of radiation. A total of 3414 embryos and 1710 plants were obtained from experiments carried out for 2 years. Considering the numbers of embryos and plants per 100 seeds, the Favorit F1 variety provided better results than the other two varieties. However, significantly different outcomes were not achieved with regard to irradiation doses. Embryos were observed at all doses tested. Flow cytometry analyses that were carried out on regenerated plants revealed whether the plants were diploid or doubled haploid, and molecular analyses revealed that diploids resulted from spontaneous chromosome doubling. The current findings offer significant results for spinach breeding using haploids.

scholarly journals In Vitro Gynogenesis and Flow Cytometry Analysis of the Regenerated Haploids of Black Cumin (Nigella sativa)

HortScience ◽  
2018 ◽  
Vol 53 (5) ◽  
pp. 681-686 ◽  
Author(s):  
Mohammed Elsayed El-Mahrouk ◽  
Mossad K. Maamoun ◽  
Antar Nasr EL-Banna ◽  
Soliman A. Omran ◽  
Yaser Hassan Dewir ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Nigella Sativa ◽  
Plant Production ◽  
Haploid Plant ◽  
Naphthalene Acetic Acid ◽  
Ms Medium ◽  
Flow Cytometry Analysis ◽  
Black Cumin ◽  
Haploid Plants

In vitro ovule culture could be used to generate homozygous lines through the production of haploid plants. The present study reports on in vitro regeneration and production of haploid plants through ovule cultures and identification of the regenerated haploids using flow cytometry. The ovules were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations of 6-benzyladenine (BA), kinetin (Kin), 2,4-dichlorophenoxyacetic acid (2,4-D), and naphthalene acetic acid (NAA) at 0, 0.5, 1, and 2 mg·L−1 for their gynogenesis. Among different plant growth regulators (PGRs) tested, 2,4-D at 2 mg·L−1 produced direct gynogenesis. The highest callogenesis percentage (100%) was obtained on MS medium containing 1 mg·L−1 2,4-D and 2 mg·L−1 NAA. Flow cytometry analysis was used to identify the regenerated haploids. It also confirmed gynogenic occurrence at 1 and 2 mg·L−1 2,4-D with percentages of 21.7% and 41%, respectively. Therefore, 2,4-D proved effective for the induction of haploids in black cumin. The regenerated haploids were developed on MS medium without PGRs. The obtained results of in vitro gynogenesis and haploid plant production can tremendously facilitate breeding programs of black cumin.

Effects of different genotypes and gamma ray doses on haploidization with irradiated pollen technique in watermelon (Citrullus lanatus L.)

2013 ◽  
Vol 93 (6) ◽  
pp. 1165-1168 ◽  
Author(s):  
Hatıra Taşkın ◽  
Namık Kemal Yücel ◽  
Gökhan Baktemur ◽  
Songül Çömlekçioğlu ◽  
Saadet Büyükalaca
Keyword(s):  
Gamma Rays ◽  
Irradiation Dose ◽  
Gamma Ray ◽  
Citrullus Lanatus ◽  
Embryo Rescue ◽  
Genotype 1 ◽  
Irradiated Pollen ◽  
Glass Tubes ◽  
Male Flowers ◽  
Haploid Embryos

Taşkın, H., Yücel, N. K., Baktemur, G., Çömlekçioğlu, S. and Büyükalaca, S. 2013. Effects of different genotypes and gamma ray doses on haploidization with irradiated pollen technique in watermelon ( Citrullus lanatus L.). Can. J. Plant Sci. 93: 1165–1168. Two watermelon genotypes, one commercial watermelon variety (Ustun F1) and five different doses of gamma rays coming from Co60 were tested to develop useful haploidization procedures in watermelon. For this purpose, male flowers collected a day before anthesis were irradiated with 50, 150, 200, 275 and 300 Gy doses of gamma rays, and female flowers were pollinated with irradiated pollen the next day. Seeds extracted from fruits harvested 25 d later were opened individually in a laminar flow hood. Embryos obtained via embryo rescue technique were placed in glass tubes containing CP medium with 30 g L−1 sucrose, 8 g L−1 agar, 0.08 mg L−1 B12, and 0.02 mg L−1 IAA. Sixty haploid embryos were obtained from 43 watermelon fruits in this study. Genotype 1 was found to be the most successful genotype with 3.57 haploid embryos per 100 seeds. Among tested irradiation doses, 275 Gy was better than other doses, with 5.26 haploid embryos per 100 seeds. Considered together with irradiation dose and genotypes, the maximum number of haploid embryos was obtained from Genotype 1 pollinated with 275 Gy irradiation dose, with 6.25 haploid embryos per 100 seeds.

scholarly journals Induction of Parthenogenetic Haploid Embryos after Pollination by Irradiated Pollen in Watermelon

HortScience ◽  
1994 ◽  
Vol 29 (10) ◽  
pp. 1189-1190 ◽  
Author(s):  
N. Sari ◽  
K. Abak ◽  
M. Pitrat ◽  
J.C. Rode ◽  
R. Dumas de Vaulx
Keyword(s):  
In Vitro Culture ◽  
Doubled Haploid ◽  
Chromosome Doubling ◽  
Citrullus Lanatus ◽  
Doubled Haploid Lines ◽  
Irradiated Pollen ◽  
Haploid Embryos ◽  
Haploid Plants

Parthenogenetic haploid embryos of `Crimson Sweet', `Halep Karasi', `Sugar Baby' and `Panonia F1' watermelon [Citrullus lanatus (Thunb.) Matsum. & Nakai] were obtained after pollination with γ-irradiated (200 or 300 Gy) pollen. Some globular and heart-shaped embryos were observed in fruit harvested 2 to 5 weeks after pollination. The number of embryos per 100 seeds was highest for `Halep Karasi'. After in vitro culture, 17 haploid plants were obtained and doubled haploid lines were generated after chromosome doubling using colchicine.

scholarly journals Isolation of Mycobacterium avium Subsp. Paratuberculosis in the Feces and Tissue of Small Ruminants Using a Non-Automated Liquid Culture Method

Animals ◽  
2019 ◽  
Vol 10 (1) ◽  
pp. 20
Author(s):  
Luigi De Grossi ◽  
Davide Santori ◽  
Antonino Barone ◽  
Silvia Abbruzzese ◽  
Matteo Ricchi ◽  
...  
Keyword(s):  
Mycobacterium Avium ◽  
Liquid Culture ◽  
Culture Media ◽  
Small Ruminants ◽  
Culture Method ◽  
Mycobacterium Avium Subsp ◽  
Type I ◽  
Culture Methods ◽  
Specific Pcr ◽  
Mycobacterium Avium Subsp Paratuberculosis

Paratuberculosis is a chronic disease of ruminants caused by Mycobacterium avium subsp. Paratuberculosis (MAP). Since isolation of MAP type I (S) is rarely reported in Italy, our research was aimed at isolating, by an inexpensive liquid culture manual method, this type of MAP isolates. At first, we used an ELISA to point out to serologically positive samples from five flocks. Secondly, we used a fecal direct IS900-qPCR on the ELISA positive samples, in order to detect shedder animals. Feces from IS900-qPCR positive samples were inoculated in solid and liquid culture media. IS900-qPCR was further used to test the growth of MAP isolates in liquid medium, which were further confirmed by f57-qPCR and submitted to typing by specific PCR in order to identify the MAP type. Twenty-eight samples (24 fecal and four tissutal samples) were processed by culture methods, resulting in the isolation of six type I MAP field isolates. Notably, no isolates were recovered by solid media, underlining the utility of this liquid method. Few data about this type of MAP are currently available in Italy, and further analyses should be carried out in order to study the origin and epidemiology of type I strains circulating in Italy.

Haploid Culture and Double Haploid Induction in Medicago sativa L. cv. XinJiangDaYe

2020 ◽  
Author(s):  
Bo Xu ◽  
Rina Wu ◽  
Fang Tang ◽  
Cuiping Gao ◽  
Xia Gao ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Medicago Sativa ◽  
Double Haploid ◽  
Haploid Plant ◽  
Regeneration System ◽  
Forage Crops ◽  
Improvement Programs ◽  
Medicago Sativa L ◽  
Cultivation Methods ◽  
Haploid Plants

Background: Alfalfa (Medicago Sativa), a perennial cross-pollinated plant, is one of the most important forage crops in the world with commercial value and ecological significance. However, due to the complexity of its genome, varietal improvement is difficult. Therefore, generating genetically homozygous materials have greater significance for breeding. In the current study, we aimed to identify the best tissue culture conditions to obtain haploid plants and double haploid plants.Methods: In this study, the haploid plants of alfalfa were obtained by combining tissue culture regeneration system with Flow cytometry. Different concentrations of colchicine were applied to the haploid plants using solid and liquid cultivation methods to determine the optimum conditions to obtain double haploid plants of Medicago Sativa L. cv. ‘XinJiangDaYe’. Result: Among the two colchicine cultivation methods tested, the doubling rate of regenerated plants obtained by liquid cultivation method was higher and the leaves developed under this system have the best doubling effect among the three explants tested. Optimal doubling conditions for alfalfa haploid (Medicago Sativa L. cv. ‘XinJiangDaYe’) were identified. The double haploid plant material generated from the current study could serve as a genetic resource for developing the hybrid combinations and for analyzing genetic linkage in alfalfa improvement programs.

Determination of radiosensitivity of Coffea arabica var. 'Venecia' seeds to gamma-ray irradiation.

2021 ◽  
pp. 320-325
Author(s):  
Reina Céspedes ◽  
Noel Arrieta ◽  
Miguel Barquero ◽  
Ana Abdelnour ◽  
Nielen Stephan ◽  
...  
Keyword(s):  
Seed Germination ◽  
Coffea Arabica ◽  
Gamma Ray ◽  
Raw Materials ◽  
New Technology ◽  
Mutation Breeding ◽  
Mutation Induction ◽  
Rust Disease ◽  
Sources Of Resistance ◽  
Breeding Programmes

Abstract Coffee is one of the most commercially available raw materials, being the tropical product with the highest market value in the world. In Costa Rica it is the third most important product for agricultural exports and provides the main income for many families in the country. However, coffee is under threat due to coffee leaf rust disease (CLR). Mutation breeding in coffee is a promising approach to develop new varieties resistant to CLR. As a new technology for coffee, basic tests related to mutation induction need to be done. The plant material used was Coffea arabica var. 'Venecia' seeds, with a moisture content of 27.3%. The applied irradiation doses were 0, 80, 100, 120, 140, 160 and 180 Gy. For each treatment, three replicates of 200 g were used, with a seed number range of 765-808 units per replicate. The irradiated seeds were planted on the same day. Eighty days after treatment the number of seedlings was quantified, the hypocotyl height and radicle length were measured and the opening of cotyledons was determined for each dose. The effects of the radiation doses on seed germination frequency were recorded. At the dose of 80 Gy, germination was reduced over the control by 9.65%, at 100 Gy by 34.06%, at 120 Gy by 52.76%, at 140 Gy by 60.24%, at 160 Gy by 65.56% and at 180 Gy by 75.40%. Seedling growth was affected and a delay in opening of the cotyledons was observed at higher doses. This radiosensitivity test, based on seed germination as compared with unirradiated control, revealed that the LD50 for the variety tested is in the range 100-120 Gy experimentally, and according to the regression is 125 ± 30 Gy. This dose will be used for further bulk experiments and is of great importance, because the LD50 is considered as the range where the appearance of useful mutations in breeding programmes is favoured. The establishment of these parameters is a necessary advance to continue with measurements of genetic and phenotypical parameters to implement mutation breeding in coffee looking for new sources of resistance against CLR.

Sign in / Sign up

Export Citation Format

Share Document