Сельскохозяйственная биология, 2010, № 4, с. 105-109.

УДК 636.087.7:582.998:57.033

ACCUMULATION OF XANTHOPHYLLS IN SOME SPECIES OF Tagetes L. GENUS IN THE CONDITIONS OF BELGORODSKAYA OBLAST’

M.Yu. Tret’yakov, V.N. Sorokopudov, V.I. Deineka

The authors investigated the Tagetes erecta and T. patula varieties on capability for industrial growing as source of xanthophylls used during manufacturing of feed addition in poultry keeping and in pharmaceutical industry. It was established that in the conditions of Belgorodskaya oblast’ the T. erecta variety with the most intensive orange blossoms (Siera orange, Rodos, Orange snow) were more preferential. The petals of the Siera orange variety contain the maximal quantity of xanthophylls diethers (3.7 mg/g, which is comparable with accumulation of xanthophylls during of growing in other world regions). «The useful mass» per one inflorescence in the T. erecta variety is 5-6 times as great as in the T. patula variety. The growing of the T. erecta plants is more efficient as far as the flowers blossom out on the plants practically simultaneously in contrast to T. patula the blossoming of which is greatly prolonged. The content of carotenoid in petals is reducing to the end of September in connection with lowering of average daily air temperature.

Key words: feed supplement, poultry, lutein, marigold (African).

The genus Tagetes is one of several genera (15 or 23, to opinion of different authors) included into the tribe Tageteae – the member of the subfamily Asteroideae, which is one of 11 those in the large family Compositae (Asteraceae) (1). In the New World, there are about 50 species of the genus Tagetes growing in the Americas from Argentina to Arizona (2, 3). Local populations of pre-Columbian era used different species of marigold as medicinal and ritual materials, and later - as an aromatic spice (4). At present time, Mexican tarragon (T. lucida) is particularly popular as a spice with a small narcotic effect, in the west Caucasus - the adventitious dwarf species T. signata (with finely-segmented fern-like leaves and lemon odour) and T. minuta (the spice imeretian saffron). In Russia and Ukraine, T. patula (local name - barkhatsy and chornobrivtsi) are widely distributed as ornamental flowering plants. In recent years, the traditional marigolds (T. patula) with dark red petals of a flower were largely replaced by varieties with different coloration and highly decorative appearance (T. erecta). T. tenuifolia, the rich-flowering low curb marigold variety, is highly appreciated as well.

Marigolds are annuals suitable for cultivation in both open flowerbeds and greenhouses. This is a very decorative and unpretentious plant, though, in contrast to Latin America, marigold doesn’t perform self-seeding in climatic conditions of our region. The practical aspect of interest to marigold is explained by an outstandingly high accumulation in its flowers of lutein diesters compared with other plants, which makes the genus Tagetes the major industrial source of this valuable xanthophyll (5). Lutein is the main pigment of the macula (yellow spot) - the area of eye retina performing most accurate focusing of light rays and the best perception of light stimuli (6). It has been established, that lutein provides a natural protection of eye retina from too intense light, and applying lutein in therapeutic purposes require the combined use of lutein and zeaxanthin at the ratio of approximately 4:1. The conventional marigold varieties contain no more than 8-10% zeaxanthin in petals; in the U.S., it has been patented the method of obtaining transgenic marigold with an increased ratio of zeaxanthin and lutein (7).

Marigolds are grown not only as an ornamental, medicinal plants and the lutein-containing raw materials for pharmaceutics, but also for producing the food additives for poultry – ORO-GLO (“Kemin Europa NV”, Belgium), Avizant Yellow (“Lohmann Animal Health” , Germany), ORI-Yellow (“Eramelko”, Spain), etc. The Russian analogue of these products - the feed additive Barfiz – was developed by the authors as a balanced combination of main carotenoid components (lutein and zeaxanthin) derived from marigold flowers (source of lutein) and calyxes of decorative physalis (source of zeaxanthin) (8).

The purpose of this work was studying the bioproductivity of Tagetes patula L. and T. erecta L. on the accumulation of xanthophylls in flowers in conditions of the Belgorod province.

Technique. The marigold species T. patula L. (the varieties Mandarin, Garmoniya, Oranzhevoye Plamya Tangerine, Harmony, Orange Flame) and T. erecta L. (the varieties Limonnyje, Rodos, Oranzhevyi Sneg, Siera Oranzhevyi, Kolanda Abrikosovyi and Antigua Lemon, Rhodes, Orange Snow, Sierra Orange, Colando Apricot and Antigua) were grown on two plots located in the city of Belgorod and in the Belgorod province in 2005-2007. The seeds were sown directly in soil (an extra close sowing) at 70 cm distance between rows. The microallotment experiments (area of plots 10 m²) were carried out in a 3-fold replication. The seeds of T. patula L. and T. erecta L. used in the experiment had been harvested in 2005 in the same locations. In 2007, the variety Sierra Orange was also grown on the 1 ha field of All-Russia Research and Development Institute of Medicinal and Aromatic Plants (Moscow); the sowing date - April, 24, collecting inflorescences - late July.

The content of xanthophylls (calculated into lutein equivalents) was determined by spectrophotometry (9).

To assess the influence of growth conditions on growth and development of the genus Tagetes, the morphometric analysis was performed. As biometrics criteria, the parameters affecting productivity and suitability for harvesting were used: number of inflorescences (n), mass of inflorescences and flowers (respectively, m and m1), stem height (h) and the number of plants per 1 m unit length (NL).

The statistical processing of data was performed by standard methods (determination of simple average, standard deviation and confidence interval) (10) using Microsoft Excel 2003 software.

Results. During the first stage of the research, it has been revealed that the highest content of lutein was characteristic for the varieties with orange color of flowers (a few milligrams per 1 g fresh flowers) (9). In the varieties with yellow, especially lemon-yellow color (in the experiment – the variety Lemon  T. erecta L.) the content of xanthophylls decreased by more than an order (while maintaining the only decorative function of plants). The dark-cherry spots typical for the variety Harmony T. patula is provided by anthocyanins, which are stable only under acidic conditions. The presence of anthocyanins in flower petals didn’t affect the content of xanthophylls that are sensitive to low pH. For the varieties with orange-colored flowers, the content of xanthophylls was comparable in both T. patula L. and T. erecta L., but the last one has much larger flowers, which makes T. erecta more convenient for industrial technology.

In 2006-2007, the highest productivity, level and dynamics of lutein accumulation compared with other varieties were detected in Sierra Orange, and Rhodes of T. erecta L. The sowing dates of marigold T. erecta L. in 2006: the varieties Sierra Orange and Colando Apricot – April 27 (entered a phase of flowering on July 24 and 20, respectively), Rhodes and Lemon – May 27, Orange Snow – May 30 (start of blossom - August 2, 17 and 23, resp.), and T. patula L. the variety Orange Flame - May 27 (early flowering occurred in August 2). Consequently, it took about 3 months from sowing until the first flowers, although it was a little over 2 month for the variety Rhodes at the late planting. In the cultivars of T. patula L., flowering began 2-3 weeks earlier than in T. erecta L. while the longer vegetation period. The highest average content of lutein was found in fresh flowers of the variety Sierra Orange - about 3,7 mg/g (Fig. 1), and almost the same - in Rhodes, with no fundamental differences between xanthophylls composition in these varieties (9).

Fig. 1. The content of xantophylls (calculated into lutein equivalents) in fresh petals of marigold  Tagetes L. the varieties: 1-7 – respectively, Sierra Orange, Rhodes, Orange Snow, Colando Apricot, Orange Flame (central flowers), Harmony, Lemon (Belgorod province, 2006). Note: abscissa – Variety, ordinate – Xantophylls content, mg/g

Dynamics of xanthophylls accumulation (Fig. 2) showed the significant reduce in content of carotenoids by the end of September (probably due to the decline of average daily temperature usual for Belgorod province in this period). The variation of this parameter in previous period was most likely caused by changes in moisture content in flowers. The content of xanthophylls about 4 mg/g is the excellent level even slightly exceeding surpassing the data obtained at growing marigold in other regions of the world (10-12).

Fig.2. Dynamics of xantophylls accumulation in petals of two varieties (A, 2006) and in two species (B, 2007) of marigold: 1 – Sierra Orange, 2 – Kolando Apricot, 3 – Antigua (Tagetes erecta), 4 – Tangerine (T. patula).   Note: А Б – A B abscissa – Date A: denotation under abscissa (from left to right) – June 21, Jule 1, Jule 11, Jule 21, Jule 31, August 10, August 20, August 30, September 9, September 19, September 29 B: denotation under abscissa (from left to right) – Jule 21, Jule 31, August 10, August 20, August 30, September 9, September 19 ordinate - Xantophylls content, mg/g

For marigold, the total productivity is determined not only by the level of xanthophylls accumulation, but also by mass of flowers, number of inflorescences per plant and by closeness of planting. Owing to this fact, in 2007 the authors performed the complex evaluation of productivity of T. erecta L. (the varieties Sierra Orange, Colando Apricot, Antigua) and T. patula L. (Tangerine). Seeds were sown on April 24; Sierra Orange, Colando Apricot and Antigua started flowering on, respectively, July 14, 16 and 20, Tangerine – on July 11.

In 2007, all the studied varieties maintained the regularities earlier shown in 2006. The relatively stable content of xanthophylls in flowers of Tagetes (see Fig. 2) allows harvesting its inflorescences from early blossom till the first frost. The optimal time - mid-August, when marigold has the highest proportion of flowering plants and the best content of lutein diesthers.

Flowers provide 50-60% weight of inflorescences in all varieties of T. erecta L. and T. patula L.; the "useful weight" of flowers of the first species exceeded the second one in 5-6 times. After keeping flowers in a cool dry place without light for 1 week, their mass significantly decreased and amounted to 14-17% of the initial in T. erecta L. and 23% - in T. patula L (Table 1). The maximum mass of flowers has been established in the cultivars Colando Apricot and Sierra Orange of T. erecta L., and Sierra Orange was considered the most promising variety, because the content of lutein diesters in its petals was higher by 30-50%.

The bioproductivity and some morphological indicators of Sierra Orange (T. erecta L.) and Tangerine (T. patula L.) (growing seasons 2006 and 2007, Table 2) suggest the possibility of their use as an industrial raw material for feed additives production.

At the industrial type of sowing T. erecta L. (Sierra Orange) in 2007, 1 ton of inflorescences was harvested (about 1/3 maximum possible yield).

Thus, for obtaining the marigold grown in the Belgorod region as a raw material for production of lutein-containing food additives, there should be selected the cultivars with flowers the most intensely colored orange (T. erecta L. - Sierra Orange, Rhodes, Orange Snow; T. patula L. - Tangerine). T. erecta L. and T. patula L. perform most similar bioproductivity, though the first species is more preferable for industrial cultivation - it is technologically easier to harvest the larger inflorescences which start blossoming almost simultaneously on a whole plant (flowering of T. patula L. is strongly prolonged). Besides, a productivity increases in the cultivars having the thicker inflorescences with higher value of the ratio  “mass of flowers / mass of inflorescence”.

The evaluation of the studied marigold varieties allows recommending Sierra Orange (T. erecta L.) for commercial cultivation in the Belgorod province.

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