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Yaniv, Z., D. Schafferman, M. Zur, and I. Shamir. 1996. Matthiola incana: Source of omega-3-linolenic acid. p. 368-372. In: J. Janick (ed.), Progress in new crops. ASHS Press, Arlington, VA.

Matthiola incana: Source of Omega-3-Linolenic Acid*

Zohara Yaniv, D. Schafferman, M. Zur, and I. Shamir

  1. METHODOLOGY
    1. Cultivation
    2. Lipid Extraction
    3. Direct Transesterification from Seeds
    4. Gas Chromatography of Methylated Fatty Acids
  2. RESULTS AND DISCUSSION
    1. Agronomy
    2. Oil Content and Quality
    3. Potential Yield
  3. REFERENCES
  4. Table 1
  5. Table 2
  6. Table 3
  7. Fig. 1
Current research in nutritional medicine indicates that the omega-3 fatty acids are receiving more and more attention as essential components of the human diet (Adam et al. 1986; Simopoulos 1986). The observed low incidence of arteriosclerosis and chronic inflammatory heart disease in Greenland Eskimos has been attributed to their traditional ethnic diet, consisting in great part of marine foods rich in two omega-3 fatty acids: C22:6 and C20:5.

Dietary fish oils containing omega-3 fatty acids are increasingly recommended for their antithrombic and hypolipidemic effects to persons consuming typical western diets (Phillipson et al. 1985). Additional benefits include improving immunologic function and fighting allergies (Leaf and Weber 1988). Omega-3 fatty acids from vegetable oils could provide health benefits without any concomitant intake of cholesterol (Hunter 1990).

Based on preliminary evaluations (Yaniv et al. 1991) we have chosen to investigate germplasm of Matthiola incana (Brassicaceae) with an average oil content of 20%-24% in the seeds and maximum levels of 65% omega-3-linolenic acid (omega3LA) of the total fatty acids in the oil (Ecker et al. 1992). This is one of the highest known content of omega3LA in a plant species (Table 1). A second important aspect of this vegetable oil is its quality as drying oil, due to the high content of omega3LA. Various Matthiola lines were tested and evaluated as a potential new-oil crop for dietary supplement and industrial uses.

METHODOLOGY

Cultivation

Five lines of M. incana selected from a collection of commercial lines for flower production, having a high content of omega3LA were tested. Plants were grown in the experiment stations at Bet Dagan, Ramat haGolan, and Jerusalem, Israel, representing three different climatic regions, during the 1991/92 growing season. Each line was replicated four times in a random block design. Seeds of each line were sown in small plots of 1.2 m2 consisting of four rows with 30 cm between rows. Basic fertilization was done at the time of soil preparation, at rates of 2N-2P-1K. 'Trifluralin' (2.5 kg/ha) was applied as a herbicide. The plots were irrigated until seedling establishment. Mean monthly temperatures at the three sites during the growing period (Nov. 1991 to June 1992) are presented in Fig. 1.

Observations were made on plant height, percent of fertile plants, and seed yield (number of pods/plant, number of seeds/pod, and weight of 1000 seeds). Plants were harvested at each location according to the time of full maturity. Fully mature seeds from each line were oven-dried overnight at 50°C and analyzed for oil content and fatty acid composition (Yaniv et al. 1991).

Lipid Extraction

Seeds were dried overnight at 50°C and ground into powder in a Moulinex coffee grinder; 5 g of powder was mixed with 100 cc petroleum ether (40° to 60°C), and the lipid fraction was extracted in a Soxhlet apparatus for 16 h at 60°C. The solvent was evaporated, and the lipid fraction residues were weighed (Yaniv et al. 1991).

Direct Transesterification from Seeds

Seeds (200 mg) were dried overnight at 50°C and ground into powder with a mortar and pestle, after which 0.3 cm3 of dichloromethane and 2.0 cm3 of 0.5 N sodium methoxide (MeONa) were added. The tube was heated to 50°C and shaken for 30 min. The reaction was stopped by adding 5.0 cm3 of water containing 0.1 cm3 of glacial acetic acid. The esterified fatty acids were extracted with 2.0 cm3 of petroleum ether (40° to 60°C). The clear fraction was kept at -20°C until further analysis. Samples of 2.0 mm3 were injected into the gas chromatograph for fatty acid analysis.

Gas Chromatography of Methylated Fatty Acids

A Megabore column (DB-23, 0.5 mm film thickness, 30 m x 0.54 mm, J&W Scientific) was used in a gas chromatograph equipped with a flame ionization detector (Varian model 3700 GC) and an automatic area integrator (3390A HP). The flow rate of N2 (carrier gas) was 30 cc/min and the oven temperature was 135° to 200°C, programmed at a rate of 4°C min. The following fatty acids were identified using known standards (Supelco): C16:0, palmitic; C18:0, stearic; C18:1, oleic; C18:2, linoleic; C18:3, linolenic; C20:1, eicosenoic; and C22:1, erucic acid.

RESULTS AND DISCUSSION

Agronomy

Table 2 summarizes the results of the evaluation of five lines of M. incana cultivated in the Bet Dagan (BD), Jerusalem (JM), and Ramat haGolan (RG) experiment stations during autumn 1991 to spring 1992. The best location was BD with all lines showing higher yield potential at that site. Yield per plant was by far the highest at BD (3.5 g vs. 0.9 g at JM and 0.7 g at RG). Number of pods/plant was 73.3 in BD as compared with 36.7 in JM and 26.7 in RG. Number of seeds/pod, plant height, pod length, and 1000 seed weight were also slightly higher at BD than at the two other sites. The best performing lines were ROZ 45 and ROZ 19. At BD, both lines showed the highest yield potential. Under the low temperature conditions prevailing in winter at JM and RG (Fig. 1), ROZ 45 and ROZ 19 maintained the highest yield. ROZ 45 and ROZ 19 had the highest fertility rate (close to 90%) at all sites tested.

Oil Content and Quality

Table 3 summarizes the oil content and fatty acid evaluation of the three test sites. Oil quantity ranged from 21% (V6) to 28%-29% (ROZ 19 and ROZ 46), with the best sites at Jerusalem and Ramat haGolan.

Seeds of ROZ 46 and ROZ 19 accumulated 29% oil in JM and RG, as compared to 25% at BD. Temperatures during seed maturation were lower at JM and RG than at BD (Fig. 1) and it is known that low temperatures during seed development have a positive effect on oil quantity (Canvin 1965; Yaniv et al. 1989). Difference in oil quantity of the three sites is probable due to temperature.

Our main goal is to obtain a high concentration of omega3LA (C18:3) in the seed oil. The levels obtained ranged from 50% to 60%, with ROZ 45, the highest at all three locations. The best locations were JM and BD. Temperature plays a major role in the relative concentration of unsaturated fatty acids of seed oils (Mazliak 1988), and cooler conditions usually favor the production of polyunsaturated fatty acids (Yaniv et al. 1995). When Matthiola were grown under controlled temperature in a Phytotron, the content of seed unsaturated omega3LA was 69% at 12°C night 17°C day as compared to 58% at 22°/27°C (Yaniv et al. 1992). However, in spite of the fact that the coolest temperatures prevailing during seed maturation were in RG (Fig. 1), the highest content of C18:3, was obtained at JM and BD, and did not go above 60% (Table 3). It is important to note that temperatures never dropped below 15deg.C during the maturation period (May-June). It could be that in order to induce a significant increase in the level of C18:3, maturation should occur under a much cooler temperature regime than tested by us in the three sites.

Potential Yield

A yield of 750 kg seeds/ha, calculated on the basis of 17 plants/m2, was obtained at BD. With 20% to 25% oil in the seed, this yield is equivalent to 150 liters oil/ha. A 50% content of omega3LA in the oil will yield 75 liters/ha pure omega3LA. Due to the fact that the research is in an early stage, we can't predict the profit per ha of the producte. This will be evaluated in during the second stage of the research.

REFERENCES

*Contribution No. 1521-E, 1995 series from the Agricultural Research Organization, The Volcani Center, Bet Dagan, Israel.
Table 1. Comparison of the fatty acid composition of five seed lipids.

Fatty acid composition ( %)
Palmitic 16:0 Stearic 18:0 Oleic 18:1 Linoleic 18:2 Linolenic omega318:3
Canola oil 6.0 1.0 62.0 20.0 11.0
Cocoa butter 26.4 31.0 37.7 3.8 T
Safflower oil 7.0 2.3 12.8 78.0 T
Soybean oil 15.0 T 24.0 54.0 7.0
Matthiola oil 9.0 2.0 13.0 11.0 65.0

Table 2. Yield parameters (mean of 4 replications) of Matthiola incana cultivated at three locations in Israel during the 1991/1992 growing season.

Sitez Line Plant ht. (cm) Fertility (%) Pods/plant Seeds/pod 1000 seeds weight (g) Yield/plant (g)
BD V-6 123 dy 37 b 68 c 48 b 1.9 ab 1.8 d
ROZ17 139 b 30 b 87 a 49 b 1.9 ab 3.4 bc
ROZ19 164 a 90 a 79 b 58 ab 2.0 ab 5.2 a
ROZ45 140 b 86 a 66 c 63 a 2.4 a 4.5 ab
ROZ46 132 c 84 a 66 c 55 ab 1.7 b 2.6 c
Avg 140 66 73 54 2.0 3.5
JM V-6 82 c 44 c 26 c 43 c 1.6 b 0.5 b
ROZ17 84 c 49 c 36 b 45 c 1.7 b 0.5 b
ROZ19 98 a 87 a 38 b 62 b 2.0 ab 1.1 ab
ROZ45 96 a 93 a 43 a 59 a 2.3 a 1.4 a
ROZ46 93 ab 68 b 41 a 52 b 1.8 b 0.9 ab
Avg 91 68 37 50 1.9 0.9
RG V-6 56 c 40 b 18 c 40 b 1.2 c 0.3 c
ROZ17 70 b 41 b 26 b 46 ab 1.5 b 0.4 c
ROZ19 83 a 88 a 31 a 54 a 1.7 b 0.8 b
ROZ45 82 a 92 a 27 b 52 a 2.2 a 1.4 a
ROZ46 81 a 81 a 32 a 50 a 1.6 b 0.6 bc
Avg 74 c 66 a 27 c 48 b 1.6 b 0.7 b
zBD = Bet Dagan; JM = Jerusalem; RG = Ramat haGolan
yMean separation in columns by Duncan's multiple range test, 1% level.

Table 3. Fatty acid composition of Matthiola incana seeds cultivated at three locations in Israel (4 replications).

Fatty acid composition (%)
Line C16:0 C18:0 C18:1 C18:2 18:3 % oil
Bet Dagan
V6 9.7 az 3.0 ab 18.4 b 11.9 b 56.3 a 21.1 c
ROZ17 9.5 a 2.5 c 21.1 a 15.6 a 50.7 c 22.5 b
ROZ19 8.6 b 2.6 bc 18.9 b 13.2 b 56.1 ab 24.7 a
ROZ45 8.8 b 3.1 a 17.7 b 12.8 b 57.1 a 25.3 a
ROZ46 8.9 b 3.1 a 19.3 ab 13.0 b 55.1 b 25.6 a
Jerusalem
V6 9.9 a 3.5 a 18.6 a 13.1 b 54.4 c 24.9 c
ROZ17 9.4 ab 2.7 c 16.4 b 14.2 a 56.8 b 25.7 bc
ROZ19 9.1 b 3.0 17.3 ab 14.1 a 55.9 b 28.9 a
ROZ45 9.0 b 3.2 ab 16.0 b 11.5 c 59.8 a 27.5 ab
ROZ46 9.1 b 3.3 ab 17.9 a 13.4 b 55.8 b 29.1 a
Ramat haGolan
V6 11.1 a 4.1 a 21.8 a 20.3 a 41.0 c 21.6 b
ROZ17 10.4 b 3.2 b 18.7 b 20.6 a 46.3 b 26.2 a
ROZ19 9.4 c 3.2 b 18.6 b 17.7 ab 50.4 a 28.2 a
ROZ45 9.2 c 3.3 b 18.2 b 14.9 b 53.6 a 26.0 a
ROZ46 9.1 c 3.5 b 20.0 b 15.3 b 51.4 a 28.8 a
zMean separation in columns by Duncan's multiple range test, 0.01% level.

Fig. 1. Mean temperatures (°C) measured at the Bet Dagan, Jerusalem and haGolan Experiment Stations, during the 1991/1992 growing season.

Last update August 21, 1997 aw