Avena genetic resources for quality in

human consumption (AVEQ)Partner 12 IHAR -PIB

Laboratory of Quality Evaluation of Plant Materials

Institute of Plant Breeding and Acclimatization - National Research

Institute,Radzików, PL 05-870, Poland AVEQ Final Meeting – Bucharest October 19-21, 2010

WP 7. Task 1, Sub-task 4 .Analyses for starch and dietary fibre

AVEQ Final Meeting – Bucharest October 19-21, 2010

The objective: Characterise carbohydrates - starch and dietary fibre in genetic resources of Avena.

WP 7. Task 1, Sub-task 4 .Analysis of starch - selected method

-amylase/amyloglucosidase method: ICC Standard no. 128/1and168 or AACC 76-13 or AOAC 996.11

AVEQ Final Meeting – Bucharest October 19-21, 2010

WP 7. Task 1, Sub-task 4 .Analysis of dietary fibre - selected

method

Gas chromatographic-colorimetric-gravimetric procedure

TDF = neutral sugar residues + uronic acids + Klason lignin

Principle1. Measurement of NSP as neutral sugar residues by GLC (Englyst and Cummings procedure, 1988)• starch removal by specific enzymes • precipitation of soluble polymers by ethanol• hydrolysis of polysaccharides to monosaccharides with H2SO4

• reduction and derivatisation

2. Measurement of uronic acid by colorimetry (Scott method, 1979)

3. Measurement of Klason lignin by gravimetric method as the sulphuric acid resistant residue (Theander et al.,1995)

Enzymatic-Chemical Uppsala Method (AOAC 994.13 or AACC 32-25)

III. Lignins

I. Non-starch polysaccharides & oligosaccharides

Cellulose Non-cellulosic polysaccharides

II. Analogous Carbohydrates

Hemicelluloses• arabinoxylans• arabinogalactans• β-glucansPectinsGumsMucilages

IV. Substances associated with NSP &

lignin complex in plants

WaxesPhytates

CutinSuberin

SaponinsTannins

Polyfructoses• inulin• oligofructansGalactooligosugars

Resistant starchSynthesized carbohydr.Indigestible dextrins

Dietary Fibre Constituents

AVEQ Final Meeting – Bucharest October 19-21, 2010

WP 7. Task 1, Sub-task 4 .Analyses for starch and dietary fibre

AVEQ Final Meeting – Bucharest October 19-21, 2010

Material analysed from the year 2008- 60 samples of selected accessions from trial in

Bulgaria- 61 samples of selected accessions from trial

in Estonia- 2 samples of wild accessions from trial in

Estonia- 6 samples of wild accessions from trial in

Poland- 53 samples of standards from trial in Estonia

Total no of accessions: 182

WP 7. Task 1, Sub-task 4 .Analyses for starch and dietary fibre

Range 51.9 – 64.1%; avg – 56.3%; CV – 4.3%

I set - selected 60 accessions from Bulgaria trial

WP 7. Task 1, Sub-task 4 .Analyses for starch and dietary fibre

NSP - range 6.1–14.1%avg - 8.8 %; CV - 13%

5.0

14.1

Uronic acids - range 0.14–0.32%avg – 0.23%; CV - 18%

6.1

2.6

TDF - range 9.4–18.2% avg - 12.0%; CV - 10%

Lignin - range 2.6–5.0%avg - 3.3%; CV - 13%

I set - selected 60 accessions from Bulgaria trial

WP 7. Task 1, Sub-task 4 .Analyses for starch and dietary fibre

Range 48.9 – 64.9%; avg – 58.7%; CV – 5.9%

II set - selected 61 accessions from Estonia trial

WP 7. Task 1, Sub-task 4 .Analyses for starch and dietary fibre

NSP - range 6.5–11.8%avg - 8.2 %; CV - 13%

Uronic acids - range 0.12–0.24%avg – 0.17%; CV - 16%

Lignin - range 1.8–4.4%avg - 2.8%; CV - 22%

II set - selected 61 accessions from Estonia trial

TDF - range 9.0–15.8% avg - 11.2%; CV - 12%

11.8

6.5

1.8

4.4

WP 7. Task 1, Sub-task 4 .Analyses for starch and dietary fibre

NSP - range 6.5–11.8%avg - 8.2 %; CV - 13%

Uronic acids - range 0.12–0.24%avg – 0.17%; CV - 16%

Lignin - range 1.8–4.4%avg - 2.8%; CV - 22%

II set - selected 61 accessions from Estonia trial plus 2 wild Avena samples

TDF - range 9.0–15.8% avg - 11.2%; CV - 12%

WP 7. Task 1, Sub-task 4 .Analyses for starch and dietary fibre

Polish wild species

AVEQ Final Meeting – Bucharest October 19-21, 2010

Accession no NSP UA Lignin TDF Starch

AVE 1426 11.3 0.29 4.5 16.1 47.8

AVE 1758 10.3 0.27 4.6 15.1 48.0

AVE 2671 11.2 0.32 4.8 16.3 46.5

AVE 2804 11.2 0.30 4.6 16.0 48.3

AVE 2781 14.0 0.32 5.0 19.3 43.9

PL 52345 15.1 0.32 4.9 20.3 40.8

WP 7. Task 1, Sub-task 4 .Analyses for starch and dietary fibre -

standards Variety Origin Starch

[DM%] SD CV

Argentina ITA 58.8 0.4 0.7Auteuil FRA 59.2 0.7 1.2Belinda SWE 60.7 0.8 1.3Evora FRA 59.7 0.8 1.3Mina BGR 57.1 0.4 0.6Ivory DEU 63.5 0.2 0.4Jaak EST 60.5 1.5 2.6Saul CZE 63.0 1.3 2.1Krezus POL 62.7 2.1 3.3Genziana ITA 59.5 1.8 3.1Mures ROM 59.8 3.4 5.7

Mean value  60.4 1.2 2.0

WP 7. Task 1, Sub-task 4 .Relationship between NSP on TDF

content

AVEQ Final Meeting – Bucharest October 19-21, 2010

TDF[% DM]

NSP [% DM]

WP 7. Task 1, Sub-task 4 .Analyses for starch and dietary fibre

Plans for the next four months:• Additional standard samples (53 samples of 11 cultivars from field experiment in Estonia) at present are analyzing for dietary fibre and they should be completed within 10 day from now. • Analyses for starch and TDF (NSP only) in 70 selected accessions from trial in Poland (2008) and France, Sweden and Italy (2009).

AVEQ Final Meeting – Bucharest October 19-21, 2010

This work has been carried out by : • Anna Fraś• Olga Jablonka• Danuta Boros

With appreciable help of two technicians:• Magdalena Bodzon• Monika Skonieczna

WP 7. Task 1, Sub-task 4 .Analyses for starch and dietary fibre

AVEQ Final Meeting – Bucharest October 19-21, 2010