Abnormal Seedlings Emerged during Embryo Rescue and Its Remedy for Seedless Grape Breeding

Eighteen seedless grape genotypes differing in ripening season (early, mid and late) and in seed trace size (small, medium and large) were harvested at 6, 10, 14, 18 and 22 weeks past bloom (wpb). Using embryo rescue techniques it was studied if embryo do abort as the fruit matures and what percent embryos remain viable at later stages. The size of seed trace was also investigated to determine its influence on embryo viability during maturation. It was found that genotype have great influence on embryo culture traits. Late maturing genotypes showed fewer rescued embryos, germinated embryos and transplantable plants than early and mid season ones. The best culture time for grape embryo rescue is 6 and 10 wpb. At these dates, the largest number of embryos, germinated embryos and transplantable plants were obtained. Genotypes with the largest ratio for seed trace weight/seed trace length (i.e., largest density) showed the greatest tendency to have the largest number of ovules with embryos, more germinated embryos and more transplantable plants. The study also showed that it is possible to recover plants from mature fruit harvested late, although at a much reduced rate.

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In-vitro embryo rescue in table-grape breeding for semiarid tropical regions of Brazil

Acta Horticulturae ◽

10.17660/actahortic.2019.1248.26 ◽

2019 ◽

pp. 179-186

Author(s):

P.C.S. Leão ◽

N.F. de Melo ◽

B.T.G. Nunes ◽

E.R. da Silva

Keyword(s):

Embryo Rescue ◽

Table Grape ◽

Tropical Regions ◽

Grape Breeding

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Breeding new seedless grape by means of in vitro embryo rescue

Genetics and Molecular Research ◽

10.4238/2013.march.26.1 ◽

2013 ◽

Vol 12 (1) ◽

pp. 859-869 ◽

Cited By ~ 9

Author(s):

W. Ji ◽

Z.-Q. Li ◽

Q. Zhou ◽

W.-K. Yao ◽

Y.-J. Wang

Keyword(s):

Embryo Rescue ◽

Seedless Grape

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Effect of culture medium, genotype, and year of cross on embryo development and recovery from in vitro cultured ovules in breeding stenospermocarpic seedless grape varieties

Australian Journal of Agricultural Research ◽

10.1071/ar07165 ◽

2008 ◽

Vol 59 (2) ◽

pp. 175 ◽

Cited By ~ 8

Author(s):

S. M. Liu ◽

S. R. Sykes ◽

P. R. Clingeleffer

Keyword(s):

Embryo Development ◽

Culture Medium ◽

Embryo Rescue ◽

Casein Hydrolysate ◽

Basal Medium ◽

Parental Genotype ◽

Genotypic Differences ◽

Embryo Recovery ◽

Seedless Grape ◽

Grape Varieties

A highly efficient embryo rescue technique using in ovulo culture is critical for maximising success when using stenospermocarpic female parents to breed new seedless grape varieties. The effects of medium composition, pollen parent, and year of cross on embryo development and recovery in cultured ovules were investigated to improve in ovulo embryo rescue from stenospermocarpic parents commonly used in breeding seedless grapes for the Australian table and dried grape industries. Increasing CaCl2 concentrations in the culture medium improved embryo recovery, but embryo emergence from ovules and germination rates were unaffected by varying FeEDTA concentrations. A casein hydrolysate supplement in the culture medium improved embryo recovery, emergence, and germination. Embryo recovery varied with parental genotype and ranged from 5% to 14% in 4 self-pollinated genotypes. Mean embryo recoveries from ovules cultured 50 days after controlled cross-pollination from berries of the pollen-sterile, stenospermocarpic variety ‘Carina’ were 36% and 26% where the pollen parents were seeded (11 crosses) and seedless (6 crosses), respectively. Embryo recovery and germination were affected by the year in which a cross was made and there was a significant cross × year interaction. The results indicated that in ovulo embryo rescue can be improved by increasing CaCl2 concentrations and incorporating a casein hydrolysate supplement in the basal medium, by exploiting genotypic differences through the use of seedless parents that yield greater proportions of rescued hybrids, and possibly by understanding environmental effects on female parents to maximise the numbers of hybrids produced.

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THE BREEDING PROGRAM OF SEEDLESS GRAPES, THE EXISTING PROBLEMS IN REPUBLIC AND A PROSPECTS OF INTRODUCING THE GEN TECHNOLOGIES

EPRA International Journal of Research & Development (IJRD) ◽

10.36713/epra3957 ◽

2020 ◽

pp. 176-181

Author(s):

Sadulla Abdullaevich Abdullaev ◽

Mukhammadlatif Marufjon o’gli Nazirov ◽

Abduvokhid Abdullaevich Bolqiev ◽

Shakhnoza Akhatullaevna Sultonova ◽

Khurshida Abdullaevna Ubaydullaeva ◽

...

Keyword(s):

Molecular Markers ◽

Breeding Program ◽

Technology Skills ◽

Gene Technology ◽

Hybrid Progeny ◽

Cross Combination ◽

Seedless Grape ◽

Seedless Grapes ◽

Grape Varieties ◽

Grape Breeding

This article discusses problems in the breeding programs of the seedless grapes, the way of introducing of the biotechnology and gene technology skills to resolve its issues, and the advantages of molecular markers to select seedlessness among the hybrid progeny of grapes. Uzbekistan is famous for its seedless grapes, but on the other hand, the grape breeding is oriented on the conventional breeding method. In other words, seedless grapes have a recessive character, a male form is using only as a pollinator, and all these factors limit to create new seedless varieties and forms. Nowadays the word grape breeders, especially seedless grape breeders use advantages of the biotechnology and gene technology skills to resolve above listed issues. As a result, except for obtaining hybrid progeny by crossing between ♂seedless × seeded ♀ that was an unchangeable algorithm of the cross combination in the traditional grape breeding. But today, researchers argued that crossing between ♂seedless × seedless ♀ is also possible to obtain a hybrid progeny through “embryo rescue technique”. Thus introducing this novel breeding program directly to grape breeding of Uzbekistan leads to an increase the diversity of the new seedless grape varieties and also for increasing the export potential of the grape industry. KEYWORDS: Seedless grape varieties, variety, cross combination, embryo culture, molecular markers, DNA.

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In vitro embryo rescue for the production of hypotetraploids after cross between hypotetraploid and tetraploid grape cultivars

Notulae Botanicae Horti Agrobotanici Cluj-Napoca ◽

10.15835/nbha48111795 ◽

2020 ◽

Vol 48 (1) ◽

pp. 503-508

Author(s):

Si-Hong KIM ◽

Joon-Ho KWON ◽

Young-Sik PARK ◽

Jae-Yun HEO

Keyword(s):

Embryo Rescue ◽

Low Frequency ◽

Cross Combination ◽

Cross Pollination ◽

Embryo Formation ◽

Seedless Grape ◽

Basal Media ◽

The Cross ◽

Grape Cultivars

Consumer demand for seedless grape with high quality and large berry has been increasing. Breeding of hypotetraploid grape was suggested as one of promising methods to satisfy it, but low frequency of hypotetraploid occurrence and low seed germination by abortive embryo were indicated as the major problem to hamper the development of hypotetraploid grape. Hence, this study was carried out to evaluate the basic efficiency of in ovulo embryo culture after the cross between hypotetraploid (‘Hanareum’) and tetraploid (‘Honey Black’ and ‘Kyoho’) grape cultivars on the establishment of hypotetraploid grapes. Embryos and plantlets were hardly obtained in ovules cultured at six after the cross pollination (WAP), but ovules inoculated at 10 WAP produced more embryos as well as plantlets regardless of cross combination. Furthermore, we found that embryo formation was not affected by the basal media in ovules cultured at six WAP, but utilization of specific medium can be more beneficial for embryo formation when ovules were cultured at 10 WAP. A total of 17 plants were obtained from ovules cultured at 10 WAP, and above 50% of plants were identified as hypotetraploid grapes. These results indicate that in vitro embryo rescue after cross pollination between hypotetraploid and tetraploid grape can enhance the efficiency for the breeding of hypotetraploid grapes.

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Improved in ovulo embryo culture for stenospermocarpic grapes (Vitis vinifera L.)

Australian Journal of Agricultural Research ◽

10.1071/ar03053 ◽

2003 ◽

Vol 54 (9) ◽

pp. 869 ◽

Cited By ~ 18

Author(s):

S. M. Liu ◽

S. R. Sykes ◽

P. R. Clingeleffer

Keyword(s):

Embryo Rescue ◽

Vitis Vinifera L ◽

Embryo Survival ◽

Recovery Rates ◽

Continuous Growth ◽

Embryo Recovery ◽

Plantlet Formation ◽

Seedless Grape

In ovulo embryo rescue techniques have been used to recover new hybrids from seedless × seedless grape crosses. This study was conducted to increase efficiency by investigating effects of genotype, medium, and ovule removal age on ovule elongation, embryo recovery, growth, and plantlet formation. Ovules from self-pollinated berries of seedless varieties Sunmuscat, Merbein Seedless, and Marroo Seedless were cultured at 30, 43, 60, and 70 days after flowering (DAF) in a range of media, some of which were supplemented with gibberellic acid (GA3) and indole-3-acetic acid (IAA). The effect of activated charcoal (AC) in media on rescued embryos was also investigated. Ovules exhibited continuous growth in vivo and in vitro. The most vigorous growth was observed for ovules cultured at 30 and 43 DAF, but more embryos were recovered from ovules cultured at 60 and 70 DAF. Ovule growth and embryo production in vitro were improved in Bouquet and Davis (BD) and Nitsch and Nitsch (NN) media. Supplementation with GA3 increased embryo recovery rates. Highest embryo recovery rates were 18.1%, 9.6%, and 12.2% for Sunmuscat, Merbein Seedless, and Marroo Seedless, respectively, when ovules were excised and cultured at 60 or 70 DAF in either BD or NN media. In vitro embryo survival and plantlet formation were higher for torpedo-shaped embryos, and improved greatly in 6-benzyladenine (BA)-supplemented woody plant (WP) medium containing 0.3% AC. Embryo recovery was improved by excising and culturing ovules at 60 DAF in BD or NN media and then by transferring embryos to WP medium supplemented with BA and AC.

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Embryo Rescue of Cold-hardy Table Grapes

HortScience ◽

10.21273/hortsci15850-21 ◽

2021 ◽

pp. 1-8

Author(s):

Laise S. Moreira ◽

Matthew D. Clark

Keyword(s):

Embryo Rescue ◽

Casein Hydrolysate ◽

Basal Medium ◽

Sampling Time ◽

Breeding Program ◽

The Other ◽

Media Composition ◽

Significant Difference ◽

Cold Hardy ◽

Grape Breeding

Seedlessness is one of the most desirable traits for table and raisin grapes. Stenospermocarpic cultivars are desirable because they have large berries with superior quality. Embryo rescue techniques have been widely used to get progeny seedling populations from crosses using seedless mother plants. Selection of the female parent, sampling time, and the growth medium are the most crucial to the success of this technique. This study investigated the effect of best sampling time and media composition on embryo rescue efficiency in a cold-hardy hybrid grape breeding program. We sampled ovules 5 to 9 weeks after flowering, and we tested four media compositions. The greatest percentages of embryo germination and normal seedlings were obtained when ovules were harvest at 8 weeks after flowering, indicating that it is suitable to harvest ovules at veraison, when the extraction of ovules is easier as a result of softer berry flesh. For the media composition experiment, all ovules were harvested at 8 weeks after flowering. Nitsch & Nitsch culture medium had very low germination, and the resulting seedlings performed the lowest compared with the other treatments. Lloyd & McCown Woody Plant Basal Medium (WPM) increased the number of embryos germinated significantly, and a number of normal seedlings and plantlets developed. Although there was no significant difference among the other three media containing WPM supplemented with different doses of plant regulators, the WPM Plus medium [with cytokinin (6-benzlaminopurine), indole-3-butyric acid, gibberellin, and casein hydrolysate] promoted the greatest percentage of established plants (46.98%). Therefore, the 8-weeks-after-flowering harvest time and the WPM Plus medium were selected for use in the embryo rescue protocol at the University of Minnesota grape breeding program.

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Embryo Rescue and Moleclar Marker-Assisted Selection of Hybrid Seedless Grape

10.21203/rs.3.rs-1185496/v1 ◽

2021 ◽

Author(s):

Yong Wang ◽

Yuling Li ◽

Feng Sun ◽

Guohong Wu ◽

Aizezi Sulaiman ◽

...

Keyword(s):

Molecular Markers ◽

Embryo Rescue ◽

Female Parent ◽

Breeding Cycle ◽

Phenotypic Traits ◽

Hybrid Female ◽

Hybrid Offspring ◽

Hybridization Rate ◽

Seedless Grape ◽

Low Efficiency

Abstract Seedless grapes play an important role in fresh food and dry production. New varieties breeding by hybridization with seedless varieties as female parents is the most effective way to cultivate seedless varieties. However, the embryos of Seedless varieties can not develop normally, so it is difficult to obtain hybrid offspring as hybrid female parent. Moreover, grape is a perennial tree species with highly heterozygous genes, with long breeding cycle and low efficiency. In this study, embryo rescue technology was used to cultivate hybrid offspring by crossing with ‘Ruby Seedless’ as female parent and ‘Hongqitezao’ as male parent, so as to solve the problem that seedless varieties can not be female parent; and molecular technology was used to carry out assisted breeding research to solve the problems of long cycle and low efficiency. TP-M13-SSR technique was used to carry out authenticity breeding. SCAR marker SCF27-2000 was used to detect the seedless traits of hybrid plants, phenotypic traits was used to verify the results of molecular markers, and Seedless trait-related SSR markers VMC7F2, VrSD10 and P3_VvAGL11 was used to detect and verify the genotypes of individual plants with inconsistent detection results by the two methods. In this study, a total of 384 hybrid offspring were finally obtained, and the hybridization rate was 84.43%. A total of 163 fruit-bearing plants were identified, and the phenotypes of their seeds were identified. The coincidence rate of genotypic and phenotypic analyses was 93.88%. Additionally, 305 F1 plants were detected using the SCF27-2000 marker, and the abortion rate was 64.92%. We speculate that the inconsistent results were caused by parthenocarpy, SCF27 marker limitation, among other factors. Overall, this study shows that embryo rescue is an effective method for breeding seedless grape cultivars, and the application of molecular markers could facilitate the early identification of hybrid traits,and improve breeding efficiency.

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New Seedless Table Grape Cultivars for the South

HortScience ◽

10.21273/hortsci.35.4.561a ◽

2000 ◽

Vol 35 (4) ◽

pp. 561A-561

Author(s):

J.R. Clark ◽

J.N. Moore

Keyword(s):

United States ◽

The United States ◽

Table Grape ◽

The South ◽

Cultural System ◽

Seedless Grape ◽

Disease Region ◽

Large Clusters ◽

Grape Cultivars ◽

Grape Breeding

Two new seedless grape cultivars were released in 1999 from the grape breeding program at the Univ. of Arkansas. `Jupiter' is the fifth release from the program. `Jupiter' is blue-fruited, has large berries, non-slipskin texture, and a mild muscat flavor. Yields of `Jupiter' were very good in replicated trials, and hardiness is also adequate for production in all areas of the South. `Jupiter' ripens 5 days later than `Venus', but earlier than `Mars' or `Reliance'. `Neptune' is the sixth release and first white-fruited cultivar from the program. It has medium-sized berries, large clusters, non-slipskin texture, and a mild, fruity flavor. Yields of `Neptune' were moderate in replicated trials. `Neptune' ripens 17 days later than `Venus' and 3 days earlier than `Mars'. Both cultivars were developed and evaluated with a commercial cultural system including routine fungicide applications, and fungicides will be required to reliably produce these cultivars. Neither of these cultivars has been tested in a Pierce's Disease region of the United States, and it is not anticipated that either will have resistance to this disease.

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