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3 Biotech

, 10:4 | Cite as

New disease-resistant, seedless grapes are developed using embryo rescue and molecular markers

  • Shasha Li
  • Zhiying Li
  • Yanan Zhao
  • Jiong Zhao
  • Qiangwei Luo
  • Yuejin WangEmail author
Original Article
  • 24 Downloads

Abstract

Improving the operation of embryo rescue technology is a highly efficient way to breed new high-quality cultivars of seedless, disease-resistant grapes when using a stenospermocarpic Vitis vinifera L. (female parents) hybrid with Chinese wild Vitis (male parents). In this study, sampling time had a significant effect on embryo recovery. Four crosses were investigated to improve the embryo rescue efficiency by determining the best sampling time, which was found to be 40, 50, 55, and 57 days after pollination (DAP) for ‘Flame seedless’ × ‘Ruby seedless’, ‘Kunxiang seedless’ × ‘Flame seedless’, ‘Ruby seedless’ self-pollination and ‘Ruby seedless’ × ‘Flame seedless’, respectively. The highest percentage embryo germination ranged from 60.83 to 89.10% for four cross-combinations when the embryo was germinated on woody plant medium (WPM), with 1.0-μM thidiazuron (TDZ, a potent cytokinin). For 11 cross-combinations, the embryo recovery rate varied from 4.8 to 29.6% and the plant development rate varied from 17.1 to 78.9%. ‘Ruby seedless’ was the best female parent followed by ‘Flame seedless’. The Grape Seedless gene Probe 1 (GSLP1) and SCF27 molecular markers were used to assist the identification of the seedless traits of the 388 progeny. A total of 309 plantlets were amplified, with the specific bands at 569-bp and 2k-bp being preliminarily considered as seedless. The S382-615 marker was used to identify the downy mildew resistance of the ‘Ruby seedless’ × ‘Beichun’ cross. Two F1 progeny were amplified, with the specific band at 615-bp being identified as downy mildew resistance.

Keywords

Grapevine Seedlessness Disease resistant Embryo rescue Molecular marker-assisted selection 

Abbreviations

IAA

Indole-3-acetic acid

6-BA

6-Benzyladenine

MS

Murashige and Skoog (1962) medium

WPM

Woody plant medium

AC

Activated charcoal

TDZ

Thidiazuron

PGR

Plant growth regulator

DAP

Days after pollination

SCAR

Sequence characterized amplified region

RAPD

Random amplified polymorphic DNA

Notes

Acknowledgements

This research at the State Key Laboratory of Crop Stress Biology in Arid Areas, Northwest A&F University, was supported by the earmarked fund for China Agriculture Research Systems for the grape industry (Grant No. CARS-29-yc-3). The authors specifically thank Dr Alexander (Sandy) Lang from RESCRIPT Co. (New Zealand) for useful language editing.

Compliance with ethical standards

Conflict of interest

No potential conflict of interest was reported by the authors.

Supplementary material

13205_2019_1993_MOESM1_ESM.doc (1.8 mb)
Supplementary material 1: Supplementary Fig. S1. Analysis of amplification by the GSLP1 marker showing the linkage to seedlessness in parents. M. DNA ladder marker (Trans DNA 2K plus), (1) ‘Beichun’, (2) ‘Shuangyou’, (3) ‘Zixiang’, (4) ‘Centennial seedless’, (5) ‘Crimson seedless’, (6) ‘Flame seedless’, (7) ‘Kunxiang’, (8) ‘Ruby seedless’, (9) ‘Huozhouhongyu’. (+) the 569-bp specific band is present; (−) the 569-bp specific band is absent. Supplementary Fig. S2. Analysis of the amplification by the SCF27 marker showing the linkage to seedlessness in parents. M. DNA ladder marker (Trans DNA 2K plus), (1) ‘Beichun’, (2) ‘Shuangyou’, (3) ‘Zixiang’, (4) ‘Centennial seedless’, (5) ‘Crimson seedless’, (6) ‘Flame seedless’, (7) ‘Kunxiang’, (8) ‘Ruby seedless’, (9) ‘Huozhouhongyu’. (+) the 569-bp specific band is present; (−) the 569-bp specific band is absent. Supplementary Fig. S3. Amplification results for GSLP1 linked to the seedless gene in ‘Flame seedless’ × ‘Shuangyou’ hybrid progeny. M. DNA ladder marker (Trans DNA 2K plus), (1) ‘Flame seedless’, (2) ‘Shuangyou’, (3–25) the 23 hybrid progeny of ‘Flame seedless × Shuangyou’. (+) the 569-bp specific band is present, (−) the 569-bp specific band is absent. Supplementary Fig. S4. Amplification results for SCF27 linked to the seedless gene in ‘Kunxiang seedless’ × ‘Flame seedless’ hybrid progeny. M. DNA ladder marker (Trans DNA 2K plus), (1) ‘Kunxiang seedless’, (2) ‘Flame seedless’, (3–28) the 26 progeny of ‘Kunxiang seedless’ × ‘Flame seedless’. (+) the 2k-bp specific band is present; (−) the 2k-bp specific band is absent. Supplementary Fig. S5. Amplification results for SCF27 linked to the seedless gene in ‘Ruby seedless’ × ‘Flame seedless’ hybrid progeny. M. DNA ladder marker (Trans DNA 2K plus), (1) ‘Ruby seedless’, (2) ‘Flame seedless’, (3–341) the 339 progenies of the ‘Ruby seedless’ × ‘Flame seedless’. (+) the 2k-bp specific band is present; (–) the 2k-bp specific band is absent. Supplementary Fig. S6. Analysis of the amplification by the S382-615 marker showing linkage to disease-resistance in parents. M. DNA ladder marker (Trans DNA 2K plus), (1) ‘Beichun’, (2) ‘Shuangyou’, (3) ‘Zixiang’, (4) ‘Centennial seedless’, (5) ‘Crimson seedless’, (6) ‘Flame seedless’, (7) ‘Kunxiang’, (8) ‘Ruby seedless’, (9) ‘Huozhouhongyu’. (+) the 569-bp specific band is present; (−) the 569-bp specific band is absent. Supplementary Fig. S7. Amplification results for S382-615 linked to disease-resistance in ‘Ruby seedless’ × ‘Beichun’ hybrid progeny. M. DNA ladder marker (Trans DNA 2K plus), (1–34) The 34 progenies of the ‘Ruby seedless’ × ‘Beichun’. (+) the 2k-bp specific band is present; (−) the 2k-bp specific band is absent. (DOC 1978 kb)

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Copyright information

© King Abdulaziz City for Science and Technology 2019

Authors and Affiliations

  1. 1.College of HorticultureNorthwest A&F UniversityYanglingPeople’s Republic of China
  2. 2.State Key Laboratory of Crop Stress Biology for Arid Areas, College of HorticultureNorthwest A&F UniversityYanglingPeople’s Republic of China
  3. 3.Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China, Ministry of Agriculture, College of HorticultureNorthwest A&F UniversityYanglingPeople’s Republic of China
  4. 4.Research Institute of Xinjiang Grape, Melon and FruitShanshanPeople’s Republic of China

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