HU212323B - Process for ergot production - Google Patents

Abstract

FIELD OF THE INVENTION The present invention relates to a novel process for the production of marmosets by the use of Triticale as a host plant for infection with gametes by infection with Claviceps fungus. EN 212 323 A Scope of the description: 6 pages

Description

The present invention relates to a method for growing nutmeg by infecting a host plant with Claviceps fungus by using Triticale treated with a gametocide as the host plant.

Emerald rye is known to contain pharmaceutically important ergolene skeletal peptide alkaloids, such as ergotamine, ergocristine and ergocriptine, which occur naturally in alpha and beta forms. Recently, the proportion of chemically modified ergolene backbone peptide alkaloids in medicine, such as dihydro and bromine derivatives, is increasing.

There are several ways to intensify the production of rye, one of which is to breed new strains of Glaviceps, which partly increase the yield of the rye and partly increase the alkaloid content of the rye (183 700, 185 701, 185 715, 186 825, 188 190, patents). On the other hand, a number of processing techniques are known to intensify the processing of mother rye, with the aim of recovering ergolenic alkaloids more efficiently.

A solution more than 20 years ago is described in U.S. Patent No. 2,048,398. Disclosure document in the Federal Republic of Germany describing the cultivation of partially or wholly genetically modified female roses on male sterile host plants. [In cytoplasmic male sterility, the combination of cytoplasmic, cytoplasmic, genetic factors associated with the mother plant, and certain homozygous recessive alleles, and only these, ensures male sterility. During several cycles of breeding male-sterile populations as seed, the recessive allozyme homozygote may not always be exclusively provided. Individuals of heterozygous (dominant-recessive) genotype for the locus in question, and thus fertile, may mix or remain in the seed population during seed selection. This circumstance explains the practice of partial male sterility. The latter can reach 3040%: 70-60% for fertile: sterile individuals.] The main advantage of this method is that spraying infection as a consequence of male sterility is sufficient instead of costly stinging.

Hungarian Patent Publication No. 195,387 describes a more economical method than male sterile rye, wherein male fertile rye is present at a concentration of 0.5-10% by weight and 0.5-50 kg / ha at or before the infection. treated with hydrazide, thereby achieving about 95% male sterility as compared to commercially available male sterile having only 60-70% sterility. With this process - about 10 years ago - the yield of mother rye was significantly increased.

Gametocidal treatment can be performed with any known gametocide. Such gametocides are described, for example, in GB 1,603,896. U.S. Pat. It is also possible to use 2,3-dichloroisobutyric acid or maleic hydrazide, preferably in the form of a salt thereof.

In the present invention, it has been a long-time object to increase the alkaloid yield of the rye, possibly by increasing its intestinal content or yield.

The female rye was cultivated on a new host plant and it was found that by using an artificial genus of wheat and rye, i.e. Triticale, as the host plant instead of rye, or, where appropriate, barley, which is commonly used, In this way, the amount of nutmeg will be higher, based on the method of cultivation with gametocide.

In accordance with the present invention, the Triticale host plant is used as an artificial genus of wheat and rye [Strauf F. Bruno; Lexicon Biological Volume 4, p. 360, Akadémiai Publishing House, Budapest 1978, Hungary; Áipád Kiss: Triticale (1968) Agricultural Publisher], first produced by W. Rimpau as a genus hybrid, but also in Hungary pioneering work on the development of varieties suitable for public production.

Triticale has several advantages over roze. It produces a slightly wider fruit. like rye and can be grown well on weak sandy soils.

The present invention relates to a novel method of growing rye by infecting a host plant with Claviceps fungus and gametocidal treatment, characterized in that an artificial genus of wheat and rye is used as the host plant.

In a preferred embodiment of the invention, Triticale is infected and the host plant is treated with a gametocide at or before the infection.

According to the invention, the host plant is applied at least once, preferably twice, in two successive phenophases of the host plant, at a rate of 0.5 to 50 kg / ha, preferably 2 to 5 kg / ha, at a dose of 0.5-101%. preferably from 2 to 5% by weight with an aqueous solution of a gametocide. The host plant reaches the first phenophase when more than 50%, preferably 70-80%, of the ears are visible, and the second phenophase begins a few days (2-3 days) before flowering begins. It should be emphasized that the development of the plant should be uniform, as this is the only way to ensure a sufficiently effective gametocidal treatment.

The infection is carried out by spraying or stabbing, preferably after or simultaneously with the gametocidal treatment, using an aqueous suspension of Claviceps fungus.

Spore number of Claviceps mushroom aqueous suspension: 1-15 x 10 ^l2 / ha

The process of the present invention is illustrated by the following examples:

/. example

In a large-plot experiment (0.3 ha), female rye was grown on two fertile Triticale cultivars and a control, partially sterile rye, and on a fertile rye cultivar as shown in Table 1. Cultivation was carried out in accordance with standard operating practice, the increase in gametocidal yields being 195 387

HU 212 323 This was done in accordance with the Hungarian Patent Specification No. Roma. The infection was effected with the strain producing ergocriptine alfa according to U. S. Patent No. 195,226 and the method of preparation of the infectious agent described in the same patent. 6 x 10 ¹² spores / if the infectious agent was applied at levels in the host plant populations such that the first infection with 2 x 10 ¹² spores / ha of inoculum sprayed in the other host plants were applied to the plant host machine puncture for the male-sterile plant host. In the week following the first inoculation, the second inoculation was carried out with a dose of 4 x 10 ¹² spores / ha of infectious agent and the method of application mentioned above. We harvested the rye with a combine harvester.

The results of the experiment and the superiority of the new host crop are summarized in Table 1, when the new host plant was used as a novel procedure concurrently with gametocidal treatment.

Table 1

host Fertilization conditions Area (ha) Gross rye yield (kg / ha) Purity of harvester mix (%) Net rye yield (kg / ha) Alpha cryptin content (mg / g) Yield (Kg / ha) Presto (Triticale) fertile 0.3 1500 98 1470 5.74 8.44 Hyclaro (rose) partially male sterile 0.3 600 95 570 6.00 3.42 Motto (rose) fertile 0.3 563 98 552 5.60 3.09 Monico (Triticale) fertile 0.3 1638 97 1590 6.80 10.81

From the table, it can be concluded that the new process is unexpected in the yields of yield and ergocriptine alpha. (New procedure: Triticale combined with gametocidal treatment.) 25

The results of OMMI (National Agricultural Certification Institute) Table 2, 3-year, small-plot experiments at different sites show, for the sake of reliability, that Triticale varieties and rye varieties do not reliably differ in their grain yields. either on an annual or annual basis. That is, the data show that in Triticale the grain yield (grain yield) does not appear to be the same as that of the rose grain, which would explain the superiority of the Triticale cultivated rye when compared to the rye cultivated rye. .

In the second example, the effect of gametocide on rowan cultivation is shown. 40

Example 2

Rye: Dankovski

Parcel size: 1 ha

The control received no gametocidal treatment, and the sting infection was performed in the usual manner with the same spore number as the other treatments.

Treatments were made with 8 L / ha Malzide (an aqueous solution containing 30% maleic hydrazide).

Time of treatments:

For single treatment: when the ears are approx. 70% visible,

For two treatments: when the ears are approx. 60% of the visible, and again 2-3 days before full flowering.

For three treatments: when 50% of the ears were visible, then when 70-75% of the ears were visible, and again one day before full flowering.

Table 2

Cereal yield data Extracted data from National Institute for Agricultural Certification (OMMI)

Rye varieties 1992 Gyula Tanya 1993 Mosonmagyaró- Castle 1993 Abaújszántó 1995 Mosonmagyaró- Castle 1995 Szombathely Tewo 4970 8570 5000 5480 5510 Presto 5210 7020 4200 5000 4920 Monico 4670 8190 4950 5800 5370 KH M 4760 Novisadi 4610 7310 3900 FA 83 3450 CT 84 4070 6650 2580 Average 4530 7550 4130 5430 5270 (X | 4534.28 7548.00 4126.00 5426.66 5266.00) Rozsfajták Rapid 5770 8550 5030 3700 6370

HU 212 323 A

Rye varieties 1992 Gyula Tanya 1993 Mosonmagyaró- Castle 1993 Abaújszántó 1995 Mosonmagyaró- Castle 1995 Szombathely Motto 5380 6970 3940 4050 5920 Dankovski Z. 5270 Small towns I. 4880 7120 1010 4350 4810 Dankovski N. 4870 Kisvárai al. 4680 6740 4210 3500 5220 Lovaszpatona 4040 7230 3920 3990 4370 Average 4980 7320 4220 3920 5340 (X ₂ 4984.28 7322.00 4222.00 3918.14 5338.23) Stats: ^ empirical 1.46 0.47 0.19 5.48 0.17 'critic (with p = 5% security level) 2.18 2.31 2.31 2.45 2.45 Annual average of Triticale varieties 4534.3 5837.0 5346.0 X ,; 92-95: 5239.1 Annual average of rye varieties 4984.3 5772.0 4628.0 X ₂ ; 92-95: 5128.1 ^ empirical 0.218 'critical (P = 5% safe level) 2.78

Infections were carried out with a spore suspension of a crystalline strain. The infection always occurred at the same time as the last treatment. spore; 4.5 x 10 ¹² / ha.

Table 3

Treatment Mill (kg / ha) Yarrow yield (kg / ha) ergocristine content (% c) yield (kg / ha) Untreated control - 230 6.8 1.56 Once treated 8 282 6.1 1.72 Treated twice 8 = 5 + 3 436 5.9 2.57 He treated me three times 8 = 4 + 2 + 2 560 6.4 3.58

Example 2 (Table 3) shows that the most favorable gametocidal treatment compared to the untreated control was 2.4 times the increase in the yield of the female rye and the increase of ca. 2.3-fold increase in alkaloid yield (ergocristine yield) in our large-plot experiment.

When comparing the data of Example 1 with the data of Example 2, it can be seen that while the gametocidal treatment (Table 3) caused an increase of 2.3 to 2.4 times the alkaloid yield and suckling yield, the Moniko Triticale The yield of gametocidal nutmeg cultivation (Table 1) is 2.8-2.9 times the yield of gametocidal nutmeg cultivation on rye (Hyclaro, Motto), while the yield of the former alkaloid is 3.1-3.5 times that of the latter.

All these show that the combined use of Triticale as a new host plant and gametocidal treatment in sow cultivation has a synergistic effect on both sow yield and alkaloid yield (yield x gut content).

The above synergistic effect is explained by the following: a) Triticalene has a typical 7-8 week growth period of mother rye for a 10-11 week growth period with gametocidal treatment. Rye without gametocide treatment for 6 weeks. Triticale takes 7 weeks for the mat to develop fully. This more than appropriate increase in breeding time of 2 weeks is one of the bases for the growth of both mother rye and alkaloid yield. During this increased breeding season, sclerocytes, meaning the average of their population, thrive and grow significantly.

Example 3

Example 3 demonstrates that gametocide-treated Triticale provides a longer passage time for growth of Claviceps fungus than gametocid4.

EN 212 323 The song is treated with rye and to take advantage of this longer period for the growth of the fungus and the growth of its gut.

The measurements of Example 3 were carried out on the plants corresponding to Example 1.

Table 4

Species Alpha Cryptin Content (% c) From the first infection to the harvest, days passed (days) Sclerocytes "1000 Eye Weight" (G) Motto 5.60 50 28.00 Hyclaro 6.00 52 20.36 Presto 5.74 67 38.60 Monico 6.80 73 42.28

Note: Harvest time is defined as the time when the intestinal content of sclerocytes reaches its maximum during post-ripening analytical measurements. (Because the maturation function of sclerocytes can be approximated by a parabola.)

(b) Triticale is a selected genus hybrid and, as such, exhibits greater morphological variability than rye. The width of the so-called "ear-zone", which plays an important role in the increase of yields, plays a crucial role in the mechanical infection (puncture infection), as a morphological feature of the host plant population. Due to the prolonged effect of gametocidal treatment on its growth (W. Perkow: Wirksubstanzen hoarfrost Pflanzenschutz- und Schadlingsbekakmpfungsmittel. Verlag Paul Parey, 1971/1982 Berlin-Hamburg), the decrease in the "ear zone" is very significant. That is, more corns receive primary infection during a puncture machine infection. As a result, the yield of nutmeg and alkaloids increases.

Example 4

No. 5 Table I below summarizes the measurements made on the plant material of the experiment described in Example 1 and on its border, which is approximately untreated with gametocide.

Table 5

Species Ear length (Cm) Measurement of the ear zone without gametocidal treatment (cm) Degree of corneal zone with gametocidal treatment (cm) Motto 9.2 ± 2.3 31.2 + 4.3 27.2 ± 4.0 Hyclaro 7.5 ± 2.0 28.7 ± 3.9 25.2 ± 3.6 Presto 11.3 ± 3.1 44.4 ± 6.9 28.3 ± 3.6

Species Ear length (Cm) Measurement of the ear zone without gametocidal treatment (cm) Degree of corneal zone with gametocidal treatment (cm) Monico 12.4 ± 3.6 43.2 ± 7.2 27.5 ± 3.1

Note: The "ear zone" is the distance between the lower ear of the lowest ear and the highest ear of the highest ear.

By the length of the ear, within a ear, is the distance between the lowest point of the lower ear and the upper point of the highest ear.

Measurements were performed in a sample area of 1 m ² with 7 replicates per breed.

It follows from the above that the superiority found in Triticale suckling roses is not due to a simple additive effect, but to Triticale being prolonged by gametocide treatment, which is inert or at least indispensable to the yield of the grain, and to Claviceps g. is a realizable advantage. Furthermore, the Triticale genus, which has a higher morphological variability, is more sensitive to morphological variability and more favorable to the rye cultivar due to hybrid gametocidal treatment. As a result of gametocidal treatment, the tapered zone of the Triticale host plant results in an increase in sultana and improved gut content.

Paragraphs (a) and (b) form the basis for the synergistic effect of Triticale and gametocidal treatment in both rye and alkaloid yields.

Claims (5)

PATIENT INDIVIDUAL POINTS A method of producing marmosets by infection of a host plant with Claviceps fungus, wherein said host is a gametocide-treated Triticale. 2. A method according to claim 1, wherein the genetically ferrous host plant is used. The method according to claim 1, wherein the genetically completely or partially male-sterile host is used. 4. The method of claim 1, wherein the host plant is infected at least once with or simultaneously with the gametocide treatment with an aqueous suspension of Claviceps fungus. 5. The method of claim 4, wherein the gametocidal treatment is by spraying and infection by spraying and / or puncture.