Part: Aspects of product quality in plant production. Quality of oil, protein and fibre crops J. Vollmann, November 2018

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1 Part: Aspects of product quality in plant production Quality of oil, protein and fibre crops J. Vollmann, November 2018

2 Oil crop species oilseed rape (canola) sunflower oil pumpkin (squash) [ soybean ] olive poppy oilseed flax (linseed) safflower novel oil crops (e.g. crambe, camelina, cuphea, lesquerella etc.) tropical / subtropical crops: sesame, cotton, peanut, coco nut, palm species...

3 Oilseed quality - the example of olive Olive (Olea europaea L.; Oleaceae) "extra-virgin olive oil": best quality, cold-pressed, first pressing only, no additives, less than 1g free fatty acids per 100 grams "virgin olive oil": less than 2 g free fatty acids ordinary virgin olive oil : less than 3.3 g per 100 grams olive oil (without the term "vergin"): oil has been refined (inferior quality, which has been improved by refining) Protected designations: Extra-virgin olive oil = "Natives Olivenöl extra"(d), "extra vergine" (I), "vierge extra" (F), "virgen extra" (Sp). (Obligatory trade specifications of the European Union since 1991, EU regulation No. 2568/91)

4 Austrian example - Rapso quality line

5 Quality of oilseed crops Oil quality: fatty acid composition, largely determined genetically Oil content: crop specific trait, influenced by agronomy and plant breeding

6 Vegetable oil systematics lipids: oils, waxes (long chain fatty acids esterified to higher alcohols), lipoids (phospholipids, glucolipids, sulpholipids, proteolipids...), emulgators (lecithin), vitamins (tocopherol) fatty acid pattern ( iodine value of vegetable oil sample...) essential fatty acids: linoleic, linolenic acid

7 Vegetable fatty acid properties

8 Vegetable oil biosynthesis fatty acid biosynthesis: FAS (fatty acid synthetase) complex on chloroplasts fatty acid modifications: desaturation, addition of functional groups such as hydroxy or epoxy groups in endoplasmatic reticulum (ER) fat (triglyceride) stored in oleosomes (oil bodies) in cotyledons

9 chloroplast Fatty acid biosynthesis (Harwood & Page, 1994)

10 Fig.: Model of oil body development in oilseeds (from: Wältermann and Steinbüchel 2005 / Vollmann and Rajcan 2009)

11 Relative fatty acid content of major vegetable oils (standard composition)

12 Relative fatty acid content of special mutants

13 Oil characteristics of crop species (summary) Brassicaceae: erucic acid (C22:1), eicosenic acid (C20:1) Asteraceae: linoleic acid (C18:2) Leguminosae: oleic acid (C18:1), linoleic acid (C18:2) linseed: linolenic acid (C18:3) Cuphea-species: medium chain fatty acids (C10:0 - C14:0)

14 Oil characteristics of crop species fatty acid patterns of crops alpha-linolenic acid (C18:3) Alpha-linolenic acid is a so-called omega-3 fatty acid.

15 Conflict over linolenic acid Linolenic acid (C18:3) is an essential fatty acid (omega-3 fatty acid). However, low-linolenic acid genotypes are promoted in soybean, rapeseed, linseed... because of the trans-fatty acid issue.

16 Selection for high oil content: The famous Illinois long term selection experiment in maize

17 Breeding for high oil content 2 ways to improve oil content: increased oil content of embryo (more oleosomes or larger oleosomes in cotyledon tissue) reduction of non-oil seed components (achene, seed hull etc.)

18 Breeding for high oil content: Seed tissue low oil Trans-sections of sunflower cotyledon cells of a low oil (upper picture) and a high oil (lower picture) genotype. o: oil body, p: protein body high oil Mantese et al. 2006

19 Breeding for high oil content examples sunflower, safflower: oil content increased from % to % through selection of genotypes with thinner achenes rapeseed: higher oil content in yellow-seeded genotypes (thinner hull) linseed: yellow seeded genotypes with reduced seed coat content contain 4-8 % more oil than genotypes with brown seed poppy: oil content increased by 3-5 % in genotypes with white seeds oil pumpkin: Schalenloser Steirischer Ölkürbis with increased oil content due to thin testa

20 Low- and highoil types of sunflower seed, rapeseed, linseed, poppy and oil pumpkin. High-oil genotypes have thin testa (seed coat) or thinner achene walls (Vollmann & Rajcan 2009)

21 Linseed / oilseed flax (Linum usitatissimum) Yellow seed variety (right): larger seed weight, higher oil content, less mucilage because of thinner seed coat than brown seeded variety

22 Oil content - environmental influences Camelina (Leindotter, false flax): 475 oil content of 10 genotypes considerably affected by environmental conditions. Higher oil content in environments with lower temperature and higher precipitation during seed filling period oil content ( g kg -1 ) GE 93 RE 93 GE 94 RE 94 environment

23 Oil content: correlation between oil content and grain yield In most cases: positive correlation between oil content and grain yield example: camelina 1993/1994, r=0.57** oil content ( g kg -1 ) r = 0.57** seed yield ( kg ha -1 )

24 Oil content - correlation between oil and protein content In nearly all species: highly negative correlation between oil and protein content example: soybean performance trial; r = oil content (g/kg) Problem: Selection for oil and protein seed protein content (g/kg)

25 Oil content - influence of planting density Response of sunflower to plant population (Robinson et al., 1980) Plants per ha Achenes per head achene weight (g) Oil content in % Head diameter in cm Plant height in cm Oil content increased at higher planting density; single plants have larger heads and a larger number of seeds per head at low density, but cannot fill so many seeds with oil. Head moisture decreased from 68 to 50% at early harvest and from 43 to 20% at later harvest, as plant density increased.

27 Oil content and quality Influence of seed position on sunflower quality (Diepenbrock, 1987) Position on the head 1000-achene weight in g Oil content in % Oleic acid in % Linoleic acid in % outer circle mid inner circle Oil content is highest at outer circles of a sunflower head due to a longer seed filling period.

29 Oil content What can a farmer do? selection of a variety with high oil content optimized planting density (too low density reduces oil content) ensure sufficient water supply during seed filling period avoid excessive nitrogen fertilization, which boosts protein content, but reduces oil sulfur fertilization of rapeseed may increase oil content optimize harvesting date and technique

30 Oil quality Rapeseed / Canola genotypes with low (less than 1%) or high (>50%) erucic acid (C22:1) content for human consumption or technical uses, respectively conventional genotypes, 0-genotypes (low erucic), 00-genotypes (=canola quality: low erucic + low content of glucosinolates) genetics: 2 major genes (allotetraploid!) influencing erucic acid content

31 Oil quality Sunflower conventional genotypes with high linoleic acid (50-70% C18:2) content high-oleic -sunflowers with oleic acid (C18:1) of up to 90 % (technical applications, high-temperature tolerant frying oil etc.); one dominant gene for high oleic acid content

32 Oil quality of safflower Genotypes with different levels of stearic (C18:0), oleic (C18:1) and linoleic (C18:2) acid content due to 3 gene loci (Knowles, 1989)

33 Oil quality Camelina variation in erucic acid content (C22:1) from below 2 % up to 6 % in progeny from a mutation breeding experiment erucic acid content (%) of individual M 3 -lines derived from M 2:3 -families a b r = 0.61 ** n = c erucic acid content (%) of M 2:3 -families

35 Oil quality Oil pumpkin (Cucurbita pepo L.) green color of oil due to chlorophyll and phaeophytine (chlorophyll-related constituent) huge differences in vitamin E (=tocopherols) content between pumpkin breeding lines from Styria (Murkovic et al., 1996) differences in linoleic acid content due to location (Schuster et al., 1983)

36 Oil pumpkin (Cucurbita pepo L.) Linoleic acid (C18:2) content (%) of 3 genotypes at different locations (N-S) Location Gießener Ölkürbis Tschermak s Ölkürbis Hybrid 3/76 Groß-Gerau, Germany Graz, Austria Nyiregyhaza, Hungary Novi Sad, Serbia Fundulea, Romania Eskisehir, Turkey Lakhmés, Tunisia (Schuster et al., 1983)

37 Oil pumpkin Content of tocopherols in seeds of different breeding lines (mg/kg) Line alphatocopherol gammatocopherol (Murkovic et al. 1996)

38 Tocopherols in general Tocopherols ( vitamin E ): alpha-, beta-, gamma-, delta-isomers in different species antioxidant effect for vegetable oil (storability / shelf life) and in human body differences in tocopherol content between species (soybean: high, pumpkin: rather low) temperature effect: high temperature = increased tocopherol content

39 Fatty acid pattern What can a farmer do??? selection of a specific variety no significant influence of agronomy on fatty acid composition environment: low temperature during seed filling period reduces saturated fatty acids and increases poly-unsaturated fatty acids (C18:2, C18:3) in many species Optimize harvesting date and reduce mechanical damage of seed to avoid free fatty acids (FFA) and rancidity! storage at reduced moisture content / temperature

40 Protein crop species soybean field pea faba bean common bean lupins lentil chickpea tropical / subtropical legumes: mung bean, cowpea etc. other legumes (alfalfa, clovers, vetches...) crops with protein meal used as a byproduct (e.g. rapeseed, sunflower and other oilseed meals)

41 Protein crop species General quality factors of food legumes for human food and animal feed

42 Source: Vollmann (2016)

43 Pulses (Grain legumes except soybean and peanut)

44 Quality of protein crops Protein quality/composition: largely determined by genetic background: amino acid composition, amino acid sequence, molecule size, 11S/7S etc. Protein content: crop specific character, influenced by agronomy and plant breeding

45 Protein quality concept of the IDEAL PROTEIN : a protein, in which no amino acid is present in a minimum concentration for nutrition (e.g. for pig fattening) biological value of a protein is given according to its distance from the ideal protein protein nutrition: plant proteins are dissolved to amino acids, di-peptides or tri-peptides, which are taken up

46 Amino acid pattern (g/160 g N) and biological value of different protein sources as related to the ideal protein for pig feeding (Boisen, 1997) amino acid BAR WHE MAI SOY PEA RAP SUN FIS Lysine Methionine Methionine + Cystine Threonine Tryptophan Isoleucine Leucine Histidine Phenylalanine Phenylalan. + Tyrosine Valine Arginine biological value

47 Protein functions Protein functions and characteristics enzyme (e.g. lipoxygenase) storage protein taste structural protein solubility (food technology parameter) gelating properties (e.g. tofu) nutritional value antinutritional value / allergenic effect...

48 Protein content environmental and genetic influences on soybean protein content protein content of a set of soybean genotypes grown in Gross Enzersdorf from ; considerable influence of season; higher protein content in seasons with high temperature and low precipitation during the seed filling period; consistent genetic differences

49 Seed protein content (g kg -1 ) Apache Ceresia 450 Proto Ultra a Year b environmental and genetic influences on soybean protein content

50 Protein content - environmental influences 475 variation of protein content in a set of soybean genotypes in 3 environments due to soil fertility differences GE PA GL Location

51 Protein content - environmental influences variation of protein content (g/kg) in a soybean field ( iso-protein lines describing field trends) Row Column

52 Protein content Nattosan control inoculation nitrogen at R1 agronomic influences and genetic differences in soybean protein content similar reaction of a set of soybean genotypes to inoculation (Bradyrhizobium japonicum) or 50 kg/ha nitrogen fertilizer with respect to enhancement of protein content Genotype Proto OAC Vision M6X-111 Apache Nebraska HM-261 Ottawa 68 Birka Sundance Leman Caruso OAC Frontier Dom M6X-89 Ma.Arrow Ceresia Dorena Polanka Leopard Labrador Kalmit Gieso Ottawa 127 Ultra LSD 0.05 mean=375.1 mean=380.0 mean=400.2 LSD 0.05 LSD Seed protein content (g kg -1 )

53 Protein correlations hp_ha 97/ normal cross high protein parent standard Plot 1 Regr 420 seed protein ,5 1,0 1,5 2,0 2,5 3,0 yield estimate Negative correlation between protein content and grain yield (example soybean)

54 Protein correlations oil content (g/kg) seed protein content (g/kg) Negative correlation between protein and oil content (soybean)

55 Rapeseed meal quality Glucosinolate content 00-genotypes: low erucic + low content of glucosinolates; less than 20 µmol of glucosinolates in fat-free meal minimizes negative effects of feeding rapeseed meal sulfur fertilization increases glucosinolate content

56 Protein content What can a farmer do? selection of a variety with high protein content (reduces grain yield most probably!!!) apply late nitrogen fertilization (e.g. fertilization after heading date, ureafertilization of rapeseed after flowering) inoculate soybean/legumes with rhizobial preparations to promote biological nitrogen fixation ( e.g. Bradyrhizobium japonicum strains)

57 BOKU Raasdorf 2005 Pischinger Watzelsdorf 2005

58 Proteingehalt (g/kg) BOKU Pischinger Results from: Protein screening 2 exp., 50 soybean genotypes, locations: BOKU (Raasdorf), Pischinger (Watzelsdorf), 2005 season.

59 Soybean seed inoculated with Bradyrhizobium japonicum nodulation bacteria

60 Comparison: Protein content (%) of crops Species % prot remark soybean trypsin inhibitors pea high starch content faba bean tannins, mycotoxins lupins alkaloids, oil content rapeseed glucosinolate sunflower linseed linamarine (glycoside) cereals 9-15

61 Amino acid composition low genetic variation in most species (selection is very difficult) no significant environmental influence cereals: high in S-amino acids soybean: best source for lysine genetic manipulation of aspartate pathway to increase lysine content (Falco et al., 1995)

62 Aspartate family biosynthetic pathway

63 Grain legumes and their ANFs (anti-nutritional factors) tannins protease inhibitors (e.g. Kunitz- trypsin inhibitor) alkaloids oligosaccharides (raffinose, stachyose) phytate (phosphorus fixed in phytic acid) lipoxygenases haemagglutinins (lectins) allergens many others

64 Grain legumes and their ANFs (anti-nutritional factors)

65 Kunitz trypsin inhibitor of soybean

66 Kunitz trypsin inhibitor of soybean Soybean Kunitz - trypsin inhibitor in SDS PAGE type of electrophoresis (absence of 21.5 kda-protein in lanes 4, 6 and 8)

67 Soybean trypsin inhibitor activity Reduced trypsin inhibitor activity in segregants free of the Kunitz protein band KTitrypsin inhibitor-activity (mg/g) KTi+ Kunitz trypsin inhibitor

68 Quality parameters of soybean oil content protein content methionine concentration trypsin inhibitors (Kunitz TI, Bowman-Birk-TI) hilum color oligosaccharides (raffinose, stachyose) activity of lipoxygenase (fat oxidation/beany flavour) linolenic acid content / saturated fatty acids tocopherol content isoflavone content

69 Food grade (left) and other types of soybean seed

70 Fibre crops Fibres: cellulose, hemicellulose, pectin, lignin Hairy fibres: cotton lint Sclerenchymatic (& xylematic) fibres ( bast fibres ): flax, hemp, stinging nettle, ramie, jute, kenaf Leaf fibres: e.g. sisal agave, fibre banana

71 fibre content fibre strength fibre length color Plant fibre quality cotton grades: color (from white to gray), foreign matter content, smoothness / roughness from harvest technical fibre characteristics

72 Flax

73 Fibre type and oilseed types of flax and their different growth habits (Linum usitatissimum L.)

74 Flax stem cross-section

75 Crosssection of a flax stem Fibrebundles in a flax stem

76 Flax fibre yield Flax (Linum usitatissimum L.) Effect of flax sowing date on fibre yield Sowing date Straw yield (kg/ha) Fibre yield (kg/ha) 26 March April April April Flax: Long-day plant (photoperiodism); late sowing causes poor vegetative development and low fibre yield due to a fixed flowering date

77 Flax fibre quality Water stress: reduction in number of fibre bundles and plant height Nitrogen fertilisation: increase in plant height, fibre length and stem diameter, effect on long fibre content not consistent Genotypes: differences in straw yield, plant height, fibre content and fibre quality

78 Stinging nettle

80 Fibre hemp

82 Fibre content of different hemp cultivars (Kriese et al., 2002)

83 Fibre hemp (Cannabis sativa L.) THC regulations in EU THC: delta-9 tetrahydrocannabinol is the psycho-active component in marijuana EU regulations (EC 1420/98): maximum THC content of 0.2 % allowed in fibre hemp varieties significant differences exist in registered varieties (Mechtler et al., 2004)

84 THC content of different hemp genotypes: range from 0.02 to 4.8 % THC (Kriese et al., 2002)

85 THC content in single plants from different low-thc cultivars grown for fibre in Austria (Mechtler et al., 2004)

Quality of oilseeds, protein crops and fibre plants

Quality of oilseeds, protein crops and fibre plants Aspects of Product Quality in Plant Production ASPECTS OF PRODUCT QUALITY IN PLANT PRODUCTION Oil and protein analytics (Practical experiments) J. Vollmann, November 2016 1. Glucosinolates 2. NIRS for