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Authors: Daniel Cristina*1,2, Alina-Gabriela Turcu 1, Vasile Mandea 1, Calina-Petruta Cornea 2 and Matilda Ciuca 1

1-National Agricultural Research and Development Institute Fundulea, N.Titulescu street; no.1, Calarasi; Fundulea; 915200; Romania

2-University of Agronomic Sciences and Veterinary Medicine of Bucharest 59 Mărăşti Boulevard, District 1 Bucharest, 011464

*danielcristina89@gmail.com

Molecular characterization of some Romanian wheat cultivars using functional molecular markers for grain weight

13th INTERNATIONALWheat Genetics Symposium23rd-28th April 2017, Tulln / Austria

National Agricultural Research andDevelopment Institute Fundulea

Food production and food security, influenced by climate changes, soil availability and accessibility, soil degradation, increase of the world population and other factors,lead to new challenges for farmers, breeders and scientist worldwide.

Wheat yield is a trait controlled by numerous genes with additive and epistatic effects that are highly interactive with the environment. Larger grains are directly relatedwith higher yield but also have favorable effects on seedling vigor and early growth, thereby promoting and stabilizing yielding ability.Large grain size has been an important trait and it is usually measured in plant breeding practice by one thousand grain weight (TGW). TGW, mainly determined by grainwidth, grain length and grain thickness, but also by grain shape and density, is a complex trait and a more detailed knowledge of its genetic control is useful for breedingprograms and breeding efficiency worldwide.

In this study we focused on the molecular characterization of 24 Romanian wheat cultivars released between 1933–2015 and 2 breeding lines, using functional markersfor genes associated with TGW components such as: TaSUS2-2B (Jiang et al. 2011), TaGW2-6A (Su et al. 2011), TaGS5-3A (Ma et al. 2015), TaTEF-7A (Zheng et al. 2014),TaCwi-A1 (Ma et al. 2012), TaGS-D1 (Zhang et al. 2014), 6-SFT-A2 (Yue et al. 2015).

INTRODUCTION

MATERIALS AND METHODSThe plant material used in this study comprised of 24 Romanian wheat genotypes released between 1933–2015 and 2 breeding lines (F628 and F132). Seeds wereharvested from yield trials performed at Fundulea (South Romania) in 2013, 2014 and 2015.Genomic DNA was extracted from two grains (grounded to fine power with a mortar and pestle). DNA extraction was performed using extraction buffer based on SDS,DNA purification using Chloroform:Isoamyl alcohol mixture (24:1 v/v) and DNA precipitation performed with absolute Ethanol.PCR reactions for analyzed genes were performed in 10 to 20 µl final volume reaction and 50-100ng DNA according to the requirements of each amplification.

Key message: Functional markers for several genes associated with grain weight revealed high genetic variability among 26 Romanian wheat genotypes for most analyzed loci, and suggest presence of other important genes.

The present work was funded through the MINISTRY OF AGRICULTURE AND RURAL DEVELOPMENT – ROMANIA, Research Project ADER116 (2015-2018).

Keywords: one thousand grain weight, TGW, functional molecular markers, wheat yield

The results are synthesized in the following table, favorable haplotypes are highlighted in red.

RESULTS AND DISCUSSION

Our results showed that some of the favorable haplotypes were present in Romanian cultivars since 1933 (A15). Analyzed genotypes showed genetic variability amongall the loci except the TaCWI-A1, where all genotypes carried the favorable haplotype. Results suggest the presence of other important genetic factors withpositive/negative influence on TGW. Also, we found, for 6-SFT-A2 gene, a NEW polymorphism in a breeding line F00628G34-1 (F628). Future research is needed toestablish if this new polymorphism is associated with desirable agronomic traits.

*FFD (Factor Form-Density) = TKW/(Length*width) (Mandea, 2016; Giura and Săulescu, 1996) *1A/1B:1R – wheat -rye translocation

ELECTROPHORETIC PATTERN

ACKNOWLEDGEMENTS

REFERENCES

Cultivar Length width FFD TKW TaGW2-6A TaSUS-2B TaTef-7A TaGS5-3A TaCwi-A1 TaGS-D1 6-SFT-A2 1A/1B:1R RHT PPD-D1 TaSAP1-A1

Diana 7.15 3.20 1.99 45.67 G Hap L Hap7A-3 T A1a A HapIII no B1A/D1A a II

Ariesan 45.20 G Hap L Hap7A-3 G A1a A HapIII no B1A/D1A a II

Lovrin 231 7.3 3.30 1.86 44.83 G Hap L Hap7A-3 T A1a A HapIII no ?/D1A a I-VI

Dropia 6.65 3.40 1.98 44.73 G Hap L Hap7A-1 G A1a A HapIII no B1B/D1A a II

Transilvania 1 7.36 3.22 1.83 43.03 G Hap L Hap7A-1 G A1a A HapIII 1B:1R B1A/D1B a III

Glosa 6.72 3.38 1.86 42.50 G Hap L Hap7A-1 G A1a A HapIII no B1B/D1A a II

Flamura 85 6.54 3.27 1.96 41.90 G Hap L Hap7A-1 G A1a A HapIII no B1B/D1A a II

Boema 1 6.61 3.37 1.88 41.83 G Hap H Hap7A-1 G A1a A no B1B/D1A a II

Ceres 6.69 3.29 1.86 40.83 G H Hap7A-1 H A1a B HapIII no B1A/D1A a III-V

Pitar 6.34 3.33 1.94 40.68 G Hap L Hap7A-1 G A1a A no ?/D1A a II

Fundulea 4 6.6 3.19 1.89 39.93 G Hap L Hap7A-1 G A1a B 1B:1R B1B/D1A b II

Pajura 6.51 3.37 1.8 39.68 A Hap L Hap7A-1 T A1a A HapIII no B1B/D1A a II

Dacia 6.06 3.29 1.97 39.40 G Hap L Hap7A-1 G A1a B HapIII no B1A/D1A b IV

Bezostaia 1 6.65 3.13 1.89 39.37 G Hap L Hap7A-1 G A1a A HapIII no B1A/D1A H III

Iulia 6.58 3.13 1.89 39.07 G Hap L Hap7A-1 T A1a B HapIII no B1A/D1A b II

Izvor 6.26 3.34 1.86 38.87 A Hap L Hap7A-1 T A1a A no B1B/D1A a IV

Litera 6.61 3.35 1.73 38.37 G Hap L Hap7A-1 G A1a A no B1B/D1A a II

F132 38.00 A Hap L Hap7A-1 T A1a A HapIII no B1B/D1A a II

Miranda 6.45 3.15 1.84 37.43 G Hap L Hap7A-1 G A1a A HapIII no B1B/D1A a II

F628 37.00 G Hap L Hap7A-1 G A1a A NEW 1A:1R B1B/D1A a I

Alex 6.86 3.26 1.64 36.90 H Hap L Hap7A-1 H A1a A HapIII no B1A/? a II

Otilia 6.01 3.26 1.88 36.77 A Hap L Hap7A-1 T A1a A no B1B/D1A a II

A15 34.90 G Hap L Hap7A-1 G A1a B HapIII no B1A/D1A b I-IV

Fundulea 133 5.97 3.00 1.93 34.67 G Hap H Hap7A-1 G A1a B HapIII no B1B/D1A b IV

Fundulea 29 6.35 3.15 1.69 33.90 H Hap L Hap7A-1 G A1a A HapIII 1B:1R ?/D1A a II

Doina 6.21 3.23 1.68 33.73 G Hap L Hap7A-3 T A1a A no B1A/D1A a II