Use of Soft X-Ray Technique to Detect Haploid Embryos in Immature Seeds of Melon

Sauton, A.; Olivier, Caroline; Chavagnat, A.

doi:10.17660/actahortic.1989.253.13

Cited by 11 publications

(7 citation statements)

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“…The effectiveness of X-ray radiography to detect and identify seeds containing immature cucumber embryos was 32% (six embryos found by X-rays out of 19 embryos found under a dissecting scope). This effi ciency was lower than that indicated by Sauton et al (1989) for melon. A total of 17 seeds were selected with X-ray radiography as containing embryos (Fig.…”

Section: Resultscontrasting

confidence: 60%

Optimization of Cucumber Doubled Haploid Line Production Using In Vitro Rescue of In Vivo Induced Parthenogenic Embryos

Claveria¹,

García-Más²,

Dolcet-Sanjuan³

2005

jashs

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ADDITIONAL INDEX WORDS. Cucumis sativus, Cucurbitaceae, parthenogenesis, haploid, DHL, pure line, hybrid ABSTRACT. Homozygous doubled haploid lines (DHLs) from new cucumber (Cucumis sativus L.) accessions could be useful to accelerate breeding for resistant varieties. DHLs have been generated by in vitro rescue of in vivo induced parthenogenic embryos. The protocol developed involves the following: 1) induction of parthenogenic embryos by pollinating with pollen irradiated with a Co 60 γ-ray source at 500 Gy; 2) in vitro rescue of putative parthenogenic embryos identifi ed by their morphology and localized using a dissecting scope or X-ray radiography; 3) discrimination of undesirable zygotic individuals from the homozygous plants using cucumber and melon SSR markers; 4) determination of ploidy level from homozygous plants by fl ow cytometry; 5) in vitro chromosome doubling of haploids; and 6) acclimation and selfi ng of selected lines. Codominant markers and fl ow cytometry confi rmed the gametophytic origin of plants regenerated by parthenogenesis, since all homozygous lines were haploids. No spontaneous doubled haploid plants were rescued. Chromosome doubling of haploid plants was accomplished by an in vitro treatment with 500 μM colchicine. Rescue of diploid or chimeric plants was shown by fl ow cytometry, prior to their acclimation and planting in the greenhouse. Selfi ng of colchicine-treated haploid plants allowed for the perpetuation by seed of homozygous lines. The high rate of seed set, 90% of the lines produced seed, facilitated the recovery of inbred lines. Despite some limiting factors, parthenogenesis is routinely used in a cucumber-breeding program to achieve complete homozygosity in one generation. Breeding for new commercial hybrid cultivars will be accelerated. DHLs are ideal resources for genomic analyses.

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Section: Resultscontrasting

confidence: 60%

Optimization of Cucumber Doubled Haploid Line Production Using In Vitro Rescue of In Vivo Induced Parthenogenic Embryos

Claveria¹,

García-Más²,

Dolcet-Sanjuan³

2005

jashs

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“…Seeds were removed from the fruits, washed with running tap water, sterilized for 20 min with 20% Clorox (1.05% NaOCl) containing 1% Tween 20, and then rinsed 3 times with sterile water. The seeds were cultured in 10 cm 9 20 mm plastic Petri dishes (*80 seeds/dish) containing 20 ml of liquid E20A medium (Sauton et al 1989) and examined periodically over a light box (Fig. 1a).…”

Section: Recovery Of Plants From Parthenogenetic Embryosmentioning

confidence: 99%

Effect of in vitro and in vivo colchicine treatments on pollen production and fruit set of melon plants obtained by pollination with irradiated pollen

Lim

Earle

2008

Plant Cell Tiss Organ Cult

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Haploid/doubled haploid (DH) technology can aid plant breeding programs by accelerating production of homozygous lines, provided enough viable DH progeny can be obtained from diverse haploid genotypes. In cases where there is a low frequency of spontaneous doubling, chromosome doubling procedures are required to achieve fecundity. We produced 63 parthenogenetic melon plantlets via pollination with c-irradiated pollen, cloned them by nodal cuttings, and tested the effects of in vitro and in vivo colchicine treatment on survival, ploidy, pollen production, and fruit recovery. The most effective procedure was in vitro exposure of 3 cm shoot tip explants to 500 mg/l colchicine for 3 h. This treatment gave 83% survival of explants and 26% conversion to diploidy. Fruit recovery rate was 60% among plants with good pollen production. In vivo exposure of the tops of young plants to 5000 mg/l for 2 and 4 h yielded some fruits but also resulted in less survival and more morphological abnormalities. Strategies for recovery of progeny from parthenogenetic melon plants are recommended. To our knowledge, this study represents the first comprehensive study of recovery of fruits and viable seeds from parthenogenetic melon plants.

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“…Use of the technique in applied breeding programs has been limited by the low percentage of haploid embryos induced, difficulty in detecting and excising these embryos, and the small numbers of haploid plantlets recovered. Savin et al (1988) and Sauton et al (1989) suggested using a soft X-ray technique to detect parthenogenetic embryos within seeds, but the equipment required is not available in many laboratories. Ficcadenti et al (1999) tested gynogenesis from unpollinated ovaries as an alternate approach to recovery of DH but did not compare it with the results from use of irradiated pollen.…”

Section: Introductionmentioning

confidence: 99%

Production of haploid and doubled haploid plants of melon (Cucumis melo L.) for use in breeding for multiple virus resistance

et al. 2003

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We have developed improved procedures for recovery of haploid and doubled haploid (DH) melon plants, using hybrids derived from crosses of lines with multiple virus resistance. Seeds formed after pollination with irradiated pollen were cultured in liquid medium for 10 days before excision of the embryos for further culture. This made it easier to identify the seeds containing parthenogenetic embryos, thereby reducing the effort required and increasing the percentage of plants recovered. The plants obtained (approximately 175) were transferred to a greenhouse for evaluation. Three fertile lines were identified, and selfed seeds were obtained for evaluating virus resistance. Flow cytometry of leaf tissues showed that two of these lines were spontaneous DH and the third was a mixoploid containing haploid and diploid cells. The other plants remained sterile through the flowering stage. Flow cytometry of 20 sterile plants showed that all were haploid. Attempts to induce chromosome doubling by applying colchicine to greenhouse-grown plants were unsuccessful. Shoot tips from the haploid plants were used to establish new in vitro cultures. In vitro treatment of 167 micropropagated haploid shoots with colchicine produced 10 diploid plants as well as 100 mixoploid plants. Pollen from male flowers that formed in vitro on the colchicine-treated plants was examined. High percentages of viable pollen that stained with acetocarmine were found not only in the diploids but also in >60% of the plants scored as mixoploid or haploid by flow cytometry. Efficient recovery of DH from hybrid melon lines carrying combinations of important horticultural traits will be a valuable tool for melon breeders.

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Use of Soft X-Ray Technique to Detect Haploid Embryos in Immature Seeds of Melon

Cited by 11 publications

References 0 publications

Optimization of Cucumber Doubled Haploid Line Production Using In Vitro Rescue of In Vivo Induced Parthenogenic Embryos

Optimization of Cucumber Doubled Haploid Line Production Using In Vitro Rescue of In Vivo Induced Parthenogenic Embryos

Effect of in vitro and in vivo colchicine treatments on pollen production and fruit set of melon plants obtained by pollination with irradiated pollen

Production of haploid and doubled haploid plants of melon (Cucumis melo L.) for use in breeding for multiple virus resistance

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