The demand for novel natural products effective against the diseases of cultivated plants, as an alternative to synthetic products which are believed to be of greater environmental impact, has stimulated research in the field of the natural products.

In particular, the need was felt for compounds of natural origin capable of replacing copper-based products which, due to their toxicity, the high dosages employed and the age-long use, are creating significant pollution problems to land and water-bearing strata.

KR Pat. 9006522 describes the use of berberine, i. e. a compound comprised in formula I, for the treatment of the pathogen responsible for apple tree canker. However, the description is confined to the effects on individual pathogen and on an individual Apple species, and with a treatment not industrially applicable.

Surprisingly, some compounds of natural origin, indicated in the present invention as piperidine benzoderivatives, have now been found to possess a marked fungicidal activity on a wide range of phytopathogenic fungi.

Hence, an object of the present invention is the use of piperidine benzoderivatives of general formula (I)

wherein represent R1 H, methoxy, ethoxy, propoxy, or, together with R2, a pentatomic dioxole group, R2 H, methoxy, ethoxy, propoxy or, together with R1, a pentatomic dioxole group, R3 H, OH, methoxy, ethoxy, propoxy or, together with R4, a pentatomic dioxole group R4 H, OH, methoxy or, together with R3, a pentatomic dioxole group R5 H, methyl, ethyl, propyl A, a saturated ring or a 10-electron aromatic system with the ring B, in which case the N atom is positively charged and/or salts and/or derivatives thereof, alone or in mixtures thereof as fungicides against phytopathogenic fungi.

It is a further object of the present invention the use as fungicides against phytopathogenic fungi of compounds of formula I wherein the ring A forms a 10- electron aromatic system with the ring B, in which case the N atom is positively charged.

Among these compounds, berberine, 9-ethoxy-13- methyl-berberine, 13-ethyl-berberine, 13-methyl- berberine, columbamine, coptisine, jatrorrhizine,

palmatine and/or worenine alone or in mixtures thereof may be mentioned.

The present invention further relates to the use as fungicides against phytopathogenic fungi of compounds having the formula I wherein the ring A is a saturated ring. Among these compounds, canadine, tetrahydropalmatine, berbine, corydaline and/or corypalmine alone or in mixture thereof may be mentioned.

It is a further object of the present invention the use as fungicides against phytopathogenic fungi of the following compounds : chelerytrine, camptothecin, chelidonine, corycavamine and/or corycavidine.

The compounds used as fungicides against phytopathogenic fungi according the present invention may be used as such or in form of salts thereof and/or derivatives, alone or in mixtures thereof.

Of course, the mixtures may also comprise all the compounds indicated in the present description.

A further subject-matter of the present invention are the fungicidal compositions active against phytopathogenic fungi which comprise at least one compound among the above mentioned ones together with carriers and/or additives commonly used in fungicidal formulations.

The compounds indicated in the present invention are easily available on the market and/or by synthesis or plant extraction with known methods.

In particular, the compounds used according to the present invention are active against fungi belonging to the following classes : Phycomycetes, Ascomycetes, Basidiomycetes, Deuteromycetes. Berberine exhibits a fungicidal activity in particular against fungi belonging to the order Oomycetales, family Peronosporaceae, genus Peronospora, species Plasmopara viticola (Betc) Beri et De Toni, genus Phytophthora ; against fungi of the order Perisporiales, family Erysiphaceae, genera Uncinula, Erysiphe, Sphaerotecha, fungi of the order

Pseudosphaeriales, family Pseudosphaeriaceae, genus Venturia and fungi of the order Helotiales, family Helotiaceae, genera Sclerotinia, Monilia/Monilinia, Botrytis/Botryotinia, fungi of the order Taphrinales, family Taphrinaceae, genus Taphrina ; against fungi of the order Ustilaginales, family Pucciniaceae, genus Puccinia ; against fungi of the order Melanconiales, family Melanconiaceae, genus Gloeosporium and against fungi of the order Hyphales, family Mucedinaceae, genus Magnaporte (Pyricularia) ; family Dematiaceae, genus Cercospora, genus Alternaria, family Tuberculariaceae, genus Fusarium.

SHORT DESCRIPTION OF THE DRAWING A single photographic reproduction is attached to the present description, showing six fruits +6 days from the treatment reported in example 4.

Berberine, whose chemical name is 5-6-dihydro-9, 10- dimethoxybenzo [g]-1, 3-benzodioxolo [5, 6-a]-quinolizinium, is a low-toxicity quaternary alkaloid present in a number of vegetal families like the Berberidacee, Papaveracee, Ranuncolacee, Rutacee. The compound is known and is also described in the Merck Index (XII Ed. hereinafter referred to as Merck Index), under N. 1192 ; the same source also mentions a method for synthesizing berberine (Kametani et col.) J. Chem. Soc. 1969, 2036).

Berberine-containing plants have been used as medicaments in the traditional medical systems of several countries, and are still being used in India, China, other Asian areas and in North America. Its therapeutical activity is explicated against malaria and several bacteria. Moreover, berberine is used as an antipyretic, anthelmintic, to expel several parasites, and it demonstrates bitter-tonic stomachic properties (Harborne JB, Baxter H., 1993).

In Western medicine, berberine is used as antibacterial agent (Bruneton, 1995 ; Reynolds, 1993).

Compounds closely related thereto exhibit a colecynetic

action, as demonstrated by Sanguinarine, which possesses acetylcholinestherase-inhibiting activities (Wichtl, 1994).

Berbine, whose chemical name is 5, 8, 13, 13a- tetrahydro-6H-dibenzo [a, g]-quinolizine is a compound described in Merck Index (N. 1195) as parent substance for naming berberine ; the reported source also mentions a synthesis, but no use thereof whatsoever.

Camptothecin, whose chemical name is (S)-4-ethyl-4- hydroxy-lH-pyrano- [3', 4' : 6, 7] indolizino [1, 2-b] quinoline- 3, 14 (4H, 12H)-dione, is also described in Merck Index (N.

1783). Useful in tumour therapy ; the substance is extracted from the stem wood of the Chinese tree Camptotheca acuminata.

Canadine, whose chemical name is 5, 8, 13, 13a. tetrahydro-9, 10-dimethoxy-6H-benzo [g]-1, 3- benzodioxolo [5, 6-a] quinolizine, described in Merck Index (N. 1785), is also a substance similar to berberine from the plant of Corydalis cava, which may be prepared by reduction of Berberine. No uses thereof are described.

Chelerythrine 1, 2-dimethoxy-12-methyl [1, 3]- benzodioxolo [5, 6-c] phenanthridinium is a substance described in Merck Index (N. 2093) obtained from root of the plant Chelidonium majus ; the same source reports a synthesis thereof and some pharmacological studies, yet not mentioning any use thereof.

Chelidonine (whose (+)-form is described as [5bR- (5ba, 6ß, 12ba)]-5b, 6, 7, 12b, 13, 14-hexahydro-13-methyl [1, 3]- benzodioxolo [5, 6-c]-1, 3-dioxolo [4, 5-i] phenanthridin-6-ol) is also described in Merck Index (N. 2095) which reports its characteristics and some studies, while not mentioning any kind of application thereof.

Columbamine (5, 6-dihydro-2-hydroxy-3, 9, 10- trimethoxydibenzo [a, g]-quinolizinium), described in Merck Index (n° 2556), is obtained from the plant of Berberis Lambertii. A synthesis from Berberine is also possible ; for this substance no application is described.

Coptisine (6, 7-dihydrobis [1, 3]-benzodioxolo [5, 6- a : 4', 5'-g] quinolizinium) is described in Merck Index (N.

2591) as obtained from the root of Copthis japonica, citing a publication related to its chemical characterization. No uses thereof are apparent, at least not from the cited source.

Corycavamine ( (R)-4, 6, 7, 14-tetrahydro-5, 14- dimethylbis [1, 3] benzodioxolo [4, 5-c : 5'. 6'-g] azecin-13 (5H)- one) is disclosed in Merck Index (n° 2607) as present in Corydalis Cava plants. The same source cites works on the structure thereof, yet no use thereof is mentioned.

Corycavidine ( (R)-5, 7, 8, 15-tetrahydro-3, 4-dimethoxy- 6, 15-dimethyl [1, 3] benzodioxolo [5, 6-e] [2] benzazecin- 14 (6H) one) is disclosed in Merck Index (n° 2608) as obtained from plant of Corydalis Cava ; the same source mentions works describing the structure and the synthesis thereof. In this case as well, no use thereof is reported.

Corydaline ( (13S-trans)-5, 8, 13, 13a-tetrahydro- 2, 3, 9, 10-tetramethoxy-13-metil-6H- dibenzo [a, g] quinolizine) is described in Merck Index (N.

2610) as an alkaloid isolated from many species of genus Corydalis. The same source cites structural, synthesis and toxicity studies.

Corypalmine (5, 8, 13, 13a-tetrahydro-2, 9, 10- trimethoxy-6H-dibenzo [g] quinolizin-3-ol) is cited in Merck Index (N. 2615) as present in Corydalis cava and in other plants belonging to genus Corydalis. The same source cites the synthesis thereof and some structural studies, yet not mentioning any use thereof.

Jatrorrhizine (5, 6-dihydro-3-hydroxy-2, 9, 10- trimethoxydibenzo [a, g] quinolizinium) is described in Merck Index (N. 5274), wherein it is indicated to be obtained from root of Jateorhiza palmata ; the same source cites structural studies, not reporting any uses thereof.

Palmatine (5, 6-dihydro-2, 3, 9, 10-tetra- methoxydibenzo [a, g] quinolizinium) according to Merck

Index (N. 7126) was isolated from root of Jateorhiza palmata ; the latter also cites the extraction process and the properties, but no use thereof.

Tetrahydropalmatine (5, 8, 13, 13a-tetrahydro-2, 3, 9, 10- tetramethoxy-6H-dibenzo [a, g] quinolizine) is cited in Merck Index (N. 9354) which also reports structure, synthesis and biosynthesis studies, mentioning the presence of a (d)-and of an (l)-form. The same source cites no use thereof.

Worenine (5, 6-dihydro-14-methylbis [1, 3] benzo- dioxolo [5, 6-a : 5', 6'-g] quinolizinium) is also described Merck Index (n° 10187) as an alkaloid similar to berberine isolated from root of Coptis Japonica. The same source cites structural studies, reporting no use thereof.

The following examples show the methods followed and the results obtained in some tests conducted to assess and quantify berberine's fungicidal activity against phytopathogenic fungi. A similar activity was found in the other compounds. The dosages indicated in the following examples relate to the quantities used in each individual treatment.

Example 1 The first test was conducted on Cucurbitaceae oidium (Erysiphe Cichoracearum DC ; Sphaerotheca Fuliginea SHL) present on gourd (Cucurbita Pepo) plants.

At a 5000 ppm dose, after a mere 24-hour time, the mycelium showed total devitalization, hence further tests with decreasing doses were conducted, with the aim of determining the minimum active dose.

The concentrations adopted were as follows : 2500 ppm, 1250 ppm, 1000 ppm, 500 ppm. Each adopted concentration, apart from the 500 ppm one, had totally devitalized the mycelium within a 48-hour time. The 500 ppm solution, despite the extremely reduced dosage thereof, showed capable of controlling the fungus, though requiring more time to exhibit its activity (about 4/5

days). The fungicidal activity is highly persistent : 14 days after the treatment, plants treated with a 1000 ppm dose exhibited no reinfection symptoms.

The following chemical compounds, commonly used against the cucurbitacea oidium, were used as reference : -Fenarimol-30 ppm -Fenbuconazole-50 ppm -Wettable Sulfur-1600 ppm None of these products, at the doses used, demonstrated a readiness and a persistence of action equalling that of the berberine. The above listed tests were repeated 5 times, always yielding the same results, within the limits of statistical errors.

Example 2 A second test was conducted on Strawberry oidium (Sphaerotheca Macularis Jacz.) ; Strawberry plants located in a nursery cultivated with biological methods were treated with 1000 ppm berberine ; after 48 hours the mycelium appeared devitalized. The reference lot, treated exclusively with 0. 3% wettable sulfur (compliant to the EEC Norms for biological productions) had instead a marked presence of active mycelium.

Example 3 The subsequent test concerned berberine's fungicidal activity towards apple scab (Venturia inaequalis [CKE] Wint = Fusicladium dendriticum [Wallr.] Fck.).

The test was conducted at the end of August in a nursery hosting the apple cultivar denominated Cripps Pink, since, due to the marked susceptibility of such cultivar to the scab, in late summer the pathogen proves difficult to control using the products known by the state of the art. Two apical treatments were carried out : --one on 08/28, at a 1000 ppm dose ; --one on 09/08, at a 1250 ppm dose.

On the same dates, two mixtures of synthetic compounds commonly used against the Apple scab were spread as reference :

--on 08/28 : 1600 ppm Mancozeb + 37 ppm Penconazole ; --and, on 09/08, 1600 ppm Mancozeb + Difeconazole 50 ppm.

On 09/14, the lower side of the first 5 fully- developed leaves located below the apical buds, well- known as the most susceptible to the pathogen, was checked ; the apical bud was not tested, since apple leaflets are scab-resistant.

The observation was conducted on 20 plants, for a total of 100 checked leaves. The yielded results were as follows : Plants treated with berberine : 68 scab spots/100 leaves = 0. 68 spots/leaf ; Plants treated with synthetic fungicides : 804 scab spots/100 leaves = 8. 04 spots/leaf.

The reading of the following tables 1 and 2 highlights the following : The good level of homogeneity of the infection within the individual theses renders the text highly significant ; The berberine-treated plants demonstrate fungal infection rates which are constantly lower with respect to those of the plants treated with synthetic fungicides, and, remarkably, the infection rate is 11. 8 times lower in plants treated with the natural product with respect to those treated with synthetic products ; The levels of infection in the plants treated with berberine are remarkably low, showing next to the initial infection rate (pre-treatment rate, visually assessed), evidence that the natural compound was capable of providing a near-total protection.

Table 1 Berberine Plant N. No. infected 1 4 2 4 3 6 4 3 5 4 6 6 7 7 8 5 9 4 10 1 11 2 12 2 13 3 14 3 15 2 16 0 17 5 18 4 19 1 20 2 Total 68 Table 2 Synthetic fungicides Plant N. No. infected 1 14 2 31 3 15 4 36 5 33 6 52 7 39 8 41 9 44 10 53 11 38 12 29 13 59 14 53 15 29 16 63 17 51 18 48 19 30 20 46 Total 804

In this case as well, berberine, besides possessing a fungicidal capacity far exceeding that of the synthetic products, demonstrated a long persistence of action, as after 10 days from the treatment no new infections emerged.

Example 4 This test was conducted on brown rot caused by Monilinia fructigena [Aderh. Et Ruhl] Honey=Sclerotinia fructigena [pers.] Aderh. Et Ruhl.

The test was conducted on 5 fruits per thesis of the Granny Smith cultivar, artificially inoculated via 3 surface scratches of an approx. length of 3 cm per fruit ; the inoculations were performed over the entire length of the scratch with a pad imbibed each time with Monilinia mycelium. The examined theses were the following : -1000 ppm/1250 ppm/2500 ppm/3000 ppm berberine ; -500 ppm procymidone (Trade name : Sumisclex- Sumitomo chem.) ; -untreated test.

The various products were applied onto the infected area immediately after the inoculation, using a small- size brush.

Later assessments provided the following results : Table 3 Assessment after 4 days from the treatment : Fruit n° Rot spot size # in cm. Thesis a (1) Untreated test 1° 2° 3° (inoculation sites) 1 3* 1 0 * presence of external 2 0. 3 0 0 mycelium 3 5* 4* 4* 4 3* 3* 1 5 5* 5* 5* Thesis b (2) Procymidone 500 ppm 1 0 0 0 2 0 1 0 3 0 0 1 4 0 0 1 5 0. 5 0. 5 0 Thesis c (3) Berberine 1000 PPM 1 1 2 0 2 0 0 0 3 1 0 0 4 0. 5 0. 5 0. 5 5 1 1 0 Thesis d (4) Berberine 1250 PPM 1 0 0 0 2 0 0 0. 5 3 0 0 0 4 0 0 0 5 0 0 0 Thesis e (5) Berberine 2500 PPM 1 0 0 0 2 0 0 0 3 0 0 0 4 0 0 0 5 0 0 0 Thesis f (6) Berberine 3000 PPM 1 0 0 0 2 0 0 0 3 0 0 0 4 0 0 0 5 0 0 0 The enclosed photographic reproduction shows the results of the assessment after 4 days from the treatment : the fruit a is the first fruit (as reported in the preceding table) of thesis 1, the fruit b is the fifth fruit of thesis 2, the fruit c is the first fruit of thesis 3, the fruit d is the second fruit of thesis 4, the fruit e is the first fruit of thesis 5, the fruit f is the first fruit of thesis 6.

Table 4 Assessment after 8 days : Thesis a (1) Untreated test 1 0 total cm 9** **The rot spot size did not 2 0 total cm 5** allow any sorting out any more 3 0 total cm 10** than the individual infection 4 # total cm 9** sites derived from the various 5 0 total cm 12** inoculations Thesis b (2) Procymidone 500 ppm 1 0 0 0 2 0 5 3 3 0 5 2 4 0 6** 5 0 8** Thesis C (3) Berberine 1000 PPM 1 9** 2 4 4 4 3 10** 4 5** 5 7** Thesis d (4) Berberine 1250 PPM 1 0 0 0 2 5** 3 6** 4 0 5** 5 0 0 0 Thesis e (5) Berberine 2500 PPM 1 0 0 0 2 0 0 4 3 0 0 5 4 0 0 0 5 0 0 0

Thesis f (6) Berberine 3000 PPM 1 0 0 0 2 0 0 0 3 0 0 5 4 0 0 0 5 0 0 0 Table 5 Assessment after 16 days Untreated reference 0 5 rotting fruits Thesis a (1) healthy fruits Procymidone 500 ppm 1 4 rotting fruits Thesis b (2) healthy fruit Berberine 1000 ppm 0 5 rotting fruits Thesis c (3) healthy fruits Berberine 1250 ppm 2 3 rotting fruits Thesis d (4) healthy fruits Berberine 2500 ppm 2 3 rotting fruits Thesis e (5) healthy fruits Berberine 3000 ppm 3 1 rotting Thesis f (6) healthy 1 starting to rot fruits The test on Monilinia highlights that berberine has, at a 1250 ppm dose, a fungicidal capacity similar to that of procymidone, whereas at 2500 ppm and 3000 ppm doses the former has a markedly superior activity versus the pathogen.

Further, the existence of a direct correlation between the dosage and the fungicidal activity is demonstrated.

Example 5 This test was conducted against Erysiphe cichoracearum on (red) radicchio (Cichorium spp) in cold greenhouse at temperatures ranging, for the entire period of assessment, between-4°C (min) and +10°C (max). This aspect is surprising, since in the cited temperature range most chemical synthesis products are ineffective against pathogenic fungi. Berberine was applied in a 1000 ppm dose ; after 6 days the mycelium showed devitalization.

Example 6 The object of this test was comparing the effectiveness of berberine towards the apple scab (Venturia Inaequalis) with that of some products commonly used in the fight against such pathogen.

The test, conducted from 19 April 2000 to 3 June 2000 in a REDCHIEF (RED DELICIOUS) apple orchard, concerned the following theses : THESIS 1-untreated THESIS 2-berberine chloride 50 g/hl (equivalent to 500 ppm) THESIS 3-berberine chloride 100 g/hl (equivalent to 1000 ppm) THESIS 4-copper sulfate pentahydrate (Cu So4 x 5 H20 25%, dose 1 Kg/hl-Mancozeb 80% dose 220 g/hl- Pyrimethanil 37. 4 dose 100 ml/hl Each lot, consisting of 5 plants, was sprayed with 3. 75 lt of solution, equivalent to about 1000 lt/ha (commonly used dose) according to the calendar reported hereinafter.

Table 6 Date* Berberine Berberine Reference 50g/hllOOg/hlProducts 2000. 02. 01 Cu So4 X 5 H20 CU S04 X 5 H20 CU S04 x 5 H20 2000. 02. 15 Cu So4 X 5 H20 Cu S04 x 5 H20 CU S04 X 5 H20 2000. 04. 19 B B- 2000. 04. 26 Mancozeb

2000.05.05 B B Mancozeb 2000. 05. 12 Pyrimethanil 2000. 05. 20 B B Pyrimethanil 2000. 06. 01 B Mancozeb 2000. 06. 07 B B Mancozeb 2000. 06. 15 B Mancozeb 2000.06.23 B B Mancozeb 2000. 06. 30 B Mancozeb *year, month, day B= treatment with berberine Total number applications : Berberine 50g/hl : 8 (+ 2 Cu So4 x 5 H20) Berberine 100g/hl : 5 (+ 2 Cu So4 x 5 H20) Reference products : 9 (+ 2 Cu So4 X 5 H2O) On 2000/05/28 the assessments on 200 leaves per thesis, and on 2000/05/28 and 2000/06/15 the assessments on fruits were conducted.

Tables 7 and 8 illustrate the yielded results.

Table 7 Assessments on fruits and leaves (2000-05-28) Scale : Class I : 2-3 spots per fruits/leaves Class II : 4-6 spots per fruits/leaves Thesis Average % of fruits Average % of leaves Scale infected D infected D UNTREATED class I 20 A 15 A class II 10 Berberine chloride class I 6 B 5. 5 B 50g/hl class 11 0 Berberine chloride class I 5 B 7 B 100g/hl class I 0 Reference class I 2 B 1 C Products class II 0 F=10. 22++ F=63. 38++ F = Fisher Test D = Duncan Test

Table 8 Assessments on fruits (2000-06-15) Scale : 1 = no attack 2 = 1-3 spots per fruit 3 = >3 spots per fruit Thesis Scale No. fruits infected Disease index UNTREATED 1 70 208 2 42 3 18 Berberine chloride 50g/hl 1 102 163 2 23 3 5 Berberine chloride 100g/hl 1 111 150 2 18 3 1 Reference products 1 118 144 2 10 3 2 Disease index = no. fruits scale 1 X 1 + no. fruits scale 2 X 2 + no. fruits scale 3X3 The tables highlight a very high activity of berberine towards apple scab.

At a dose of 50 g/hl (equivalent to 500 ppm) dispensed on a weekly basis, Berberine provided results similar to those of the traditional products, especially in fruit protection.

Remarkably, at a dose of 100 g/hl (equivalent to 1000 ppm) similar results were attained, almost halving the number of interventions with respect to the calendar commonly adopted with the traditional products, thus entailing extremely remarkable advantages regarding both the environmental impact and the costs.

Example 7

The following test was conducted to assess the effectiveness of Berberine towards the pathogen Uncinula necator, causing the vines Oidium.

The test was conducted from 2000/05/10 to 2000/08/21 in a Chardonnay grapevine. The theses provided were as follows : Table 9 Treatments TREATMENT Rate/hl Thesis A. I. Form g/hl 1 untreated 2 Berberine chloride Berberine chloride 50 (CoH, $CIN04x2H20) (Fluka) 3 Berberine chloride Berberine chloride 100 (C20Hl8ClNO4x2H2O) (Fluka) 4 Ampelomyces quisqualis AQ10 45 Reference Miclobutanil (i. a. 6. 2) Systhane SC/Xi 35-40 Products S 80% Micronised sulfur 300 Dinocap-miclobutanil Sabithane EC/Xi 60-80 (i. a. 32. 9 +7. 6)

50 g/hl and 100 g/hl are equivalent to 500 and 1000 ppm, respectively.

The calendar of the treatments developed as follows : Table 10 Applications Date* Berberine 50g/hl Berberine lOOg/hi Reference Products 2000. 05. 09 - - AQ10 2000. 05. 10 B B (shoot 15-20 cm. in length) 2000. 05. 18 - - AQ10 2000. 05. 24 B B (blossoming) 2000. 05. 31--MICLOBUTANIL (DOSE 35G/HL) 2000. 06. 07 B B - 2000. 06. 09 - - S 80% 2000. 06. 16 MICLOBUTANIL (DOSE 40G/HL) 2000. 06. 21 B B - 2000. 06. 28 MICLOBUTANIL (DOSE 40G/HL) 2000. 07. 05 B B - 2000. 07. 07--DINOCAP- MICLOBUTANIL (DOSE 80G/HL) 2000. 07. 14 B B - 2000. 07. 21 - - DINOCAP- MICLOBUTANIL (DOSE 80G/HL) 2000. 07. 24 B B - 2000. 08. 02BB- 2000. 08. 03--DINOCAP- MICLOBUTANIL (DOSE 60G/HL) AQ10 2000. 08. 08 B B - *year/month/day

B = treatment with Berberine Each 3 plant-thesis was sprayed with a quantity of solution equivalent to 600 1/ha from April 28, 2000 to May 24, 2000 and with 1000 1/ha thereof from July 6, 2000 to August 8, 2000. The assessments on leaves were conducted prior to the treatment and at the onset of ripening ; for each of the 3 plants per thesis 50 leaves, i. e., a total of 150 leaves, were assessed. The assessments on fruits were conducted on August 8, 2000 on 50 grapes per thesis, and the % of infected grapes was assessed.

The tables reported herebelow illustrate the yielded results.

Table 11 Assessments on leaves DATE untreated Berberine 50 g/hl Berberine 100 g/hl Reference Products P I P I P I P I 2000. 04. 28 no no no no no no no no 2000. 05. 10 no no no no no no no no 2000. 05. 24 no no no no no no no no 2000. 06. 07 no yes no no no no no no 2000. 06. 21 no yes no no no no no no 2000. 07. 05 no yes no no no no no no 2000. 07. 14 no yes no yes no yes no yes 2000. 07. 24 no yes no yes no yes no yes 2000. 08. 02 no yes no yes no yes no yes 2000. 08. 08 no yes no yes no yes no yes Legend : P = Phytotoxicity I = Infection

Table 12 Assessments on untreated thesis leaves DATE % of leaves area affected N. leaves affected/ n. leaves assessed 2000. 06. 07 4. 7 3/150 2000. 06. 21 6. 5 43/150 2000. 07. 05 14. 3 120/150 Table 13 Assessment on leaves (July 14, 2000) Treatment plant % of leaves area Average value D affected m UNTREATED 1 18 2 20 19. 5 A 3 20. 5 Berberine chloride 1 19. 5 50g/hl 2 18 18. 3 A 3 17. 5 Berberine chloride 1 15 100g/hl 2 15. 5 14. 8 B 3 14 Reference Products 1 0. 5 2 0. 5 0. 6 C 3 0. 8 F=734. 43++ Treatment plant N. leaves affected/Average value D n. leaves assessed m UNTREATED 1 50/50 2 50/50 50/50 A 3 50/50 Berberine chloride 1 48/50 50g/hl 2 50/50 48/50 A 3 46/50 Berberine chloride 1 47/50 100hl 2 46/50 48/50 A 3 50/50 Reference Products 1 7//50 2 6/50 7/50 B 3 9/50 F=66. 87++ F = Fisher Test D = Duncan Test Table 14 Assessments on leaves (August 8, 2000) Treatment plant % of leaves area Average value affected m D untreated 160 2 40 50 A 3 50 Berberine 1 40 chloride 2 35 38. 3 A 50 g/hl 3 40 Berberine 1 15 chloride 2 20 15 B 100 hl 3 10 Reference 1 2 Products 2 1 2 c 3 3 F=57.22++ Treatment plant N. leaves affected/Average value n. leaves assessed m D UNTREATED 1 50/50 2 50/50 50/50 A 3 50/50 Berberine 1 50/50 chloride 50g/hl 2 50/50 50/50 A 3 50/50 Berberine 1 47/50 chloride 100hl 2 48/50 48/50 A 3 50/50 Reference 1 15/50 Products 2 10/50 10/50 B 3 5/50 F= 21. 77++ F = Fisher Test D = Duncan Test

Table 15 Assessment on fruits, on August 8, 2000 Treatments plant % berries Average value infected m D UNTREATED 1 55 2 60 58. 3 A 3 60 Berberine chloride 50g/hl 1 50 2 40 41. 6 B 3 35 Berberine chloride 1 20 100 g/hl 2 10 18. 3 C 3 25 Reference Products 1 5 2 2 3 D 3 2 F=69. 42++ F = Fisher Test D = Duncan Test The results demonstrate that the effectiveness of berberine grows in function of the dosage thereof, though in order to obtain a result comparable to those of the traditional products in the initial periods of greater pathogenic virulence a calendar with closer treatments will have to be adopted, as actually performed in the standard reference thesis.

Example 8 In this thesis berberine's activity towards Taphrina deformans, causing the peach leaf curl, was assessed.

In common practice, the fight against leaf curl requires some treatments to be performed in wintertime, with products like ZIRAM and TMTD, highly noxious to man and toxic to the environment.

An intervention was performed on March 30, 2000 with undergoing infections, in order to assess berberine's curative activity.

Two treatments, at a 7-day interval therebetween, were performed at a dose of 100 g/hl berberine equivalent to 1000 ppm ; two weeks after the last treatment the average number of healthy leaves per shoot was assessed.

Table 16 Assessment April 20, 2000 Thesis Average N. of m % activity healthy leaves Untreated 1, 9 2 1. 4 1. 5 1. 7- Berberine 2. 8 2. 6 2. 8 2. 1 2. 56 51 chloride Rate of activity = Untreated plant mean-Treated plant mean <BR> <BR> <BR> ----------------------------------------.---------xlOO Untreated plant mean Results : decrease of intensity of disease of 51% in Berberine chloride thesis Phytotoxicity : absent The results yielded highlight a very high effectiveness of action, hence berberine may constitute a useful means for the ecocompatible fight against Taphrina deformans.

Example 9 The test was conducted in order to assess berberine's effectiveness towards Cercospora bieticola, a pathogen causing serious damages to sugar beet.

The lot examined had already been pretreated with Cu So4 x 5H20 (25% Cu) at a dose of 700 g/hl.

A test was performed on July 16, 2000 according to the following pattern : 1. Untreated reference 2. 100 g/hl Berberine chloride equivalent to 1000 ppm 3. 700 g/hl Cu SO4 x 5H20 Two weeks later, the % of leaves affected was assessed on 30 randomly selected plants per thesis.

The table illustrates the yielded results.

Table 17 Assessment July 29, 2000 Thesis % of leaf area % activity affected m Untreated 10 Berberine 4 60 chloride Reference 2. 5 75 Products Cu S04 x 5 H20 Rate of activity= mean of untreated plants-mean of treated plants <BR> <BR> <BR> x 100 mean of untreated plants Results : decrease of intensity of disease of 75% in Reference Product Thesis and of 60% in Berberine Chloride Thesis Phytotoxicity : absent The outcome of the test highlights how, already at 100 g/hl, berberine is capable of significantly controlling the pathogen.

Example 10 The object of this test was assessing Berberine's activity towards the fungus Plasmopara viticola, causing the Vine Peronospora.

The test was conducted on plants of the variety Sultana, particularly prone to Peronospora attacks.

The theses compared were the following : Untreated reference 100 g/hl berberine chloride, equivalent to 1000 ppm

# 1000 g/hl Cu So4 x 5H2O (Cu 25%) INTERVENTION TIMES On April 26, 2000, with the shoots 15-20 cm in length, the first treatment was performed, followed by other two at a 10-day interval thereamong ; from May 20, 2000 to June 6, 2000 the treatments were performed weekly, according to the scheme reported in table 18.

Table 18 DATE* Thesis untreated Berberine chloride Reference Product 04/24/00 B CuS04 05/01/00 CuS04 05/03/00B 05/08/00 CuS04 05/13/00B 05/15/00 CuS04 05/20/00 B 05/23/00 CuS04 05/27/00 B 05/30/00 CuS04 06/03/00 B 06/06/00 CuS04 06/10/00 B 06/13/00 CuS04

*month/day/year B= treatment with Berberine On June 12, 2000 the test was suspended due to a severe hail storm.

Each thesis consisted of two plants, and on May 20, 2000 and June 10, 2000 on 20 randomly selected shoots per thesis the rate of affected leaves and of the leaf surface affected was assessed.

The data are reported in the following table : Table 19 ASSESSMENTS ON SULTANA 20 May 2000 10 June 2000 Thesis % leaves % % % leaves % % affected leaf surface activity affected leaf surface activity affected affected untreated 80 14. 3-95 24- Berberine 66 9. 5 33. 5 78 12. 8 46. 6 chloride Farmer 35 4. 5 68. 5 40 7. 5 68. 7 Rate activity= mean of untreated plants-mean of treated plants -------------------------------------------------- x 100 mean of untreated plants Results : May 20, 2000 decrease of intensity of disease of 68. 5% in Farmer Thesis and of 33. 5% in Berberine chloride Thesis ; June 10, 2000 decrease of intensity of decrease of 68. 7% in Farmer Thesis and of 46. 6% in Berberine chloride Thesis.

Phytotoxicity : absent The thesis demonstrates berberine, already at 100 g/hl, to be active against the pathogenic agent, despite a flaring back of the infection due to adverse weather condition.

Example 11 The following test aimed at assessing berberine's activity towards Plasmopara viticola (Vine Pernospora) on the species of vine Sangiovese, the latter being of particular economic importance, due to its widespread cultivation.

The test was conducted as follows :

# Untreated reference Berberine chloride 100 g/hl, equivalent to 1000 ppm # Cu S04 x 5H20 (Cu 25%)-1000 g/hl # Plants for Thesis n. 2 APPLICATIONS : The first treatment with berberine was conducted with 20-cm long shoots on May 2, 2000 ; the subsequent treatments were applied every 10 days until August 04, 2000 for a total of 12 treatments.

The treatments with Copper sulfate (Cu So4 x 5H20) started on April 30, 2000 and protracted weekly until August 10, 2000, for a total of 15 treatments as reported in table 20 Table 20 DATE* Thesis Untreated Berberine chloride Reference Products 04/30/00 CuS04 05/02/00 B 05/07/00 CuS04 05/12/00 B 05/14/00 CuSO4 05/21/00 CuSO4 05/22/00 B 05/28/00 CuSO4 05/31/00 B 06/04/00 CuSO4 06/09/00 B 06/11/00 CuSO4 06/18/00 CuSO4 06/19/00 B 06/25/00 CuSO4 06/27/00 B 07/02/00 CuSO4 07/06/00 B 07/09/00 CuS04 07/16/00 B CuS04 07/23/00 CuS04 07/26/00 B 07/30/00CuS04 08/03/00 B 08/06/00 CuS04 08/10/00 B *month/day/year B= Treatment with Berberine On August 10, 2000 the rate of healthy leaves of 20 randomly selected shoots per thesis were counted, and the rate of leaf surface affected was calculated.

No symptom of infection appeared on the fruits.

The data are reported in the following table : Table 21 Assessment on"SANGIOVESE"August 10, 2000 Thesis % leaves affected % leaf surface % activity affected Untreated 20 8. 5 Berberine chloride 16 3. 5 58. 8 Reference Product 9 3. 2 62. 3 Rate activity= mean of untreated plants-mean of treated plants <BR> <BR> <BR> x 100 mean of untreated plants Results : decrease of intensity of disease of 62. 3% in reference product thesis and of 58. 8% in berberine chloride thesis.

Phytotoxicity : absent The test demonstrates Berberine, at a mere 100 g/hl and with treatments far apart, to have, with respect to copper oxychloride, an activity substantially similar to the latter and to be a worthy ecocompatible alternative

against Plasmopara viticola.

Example 12 This test aimed at assessing berberine's effectiveness towards the collar rot of vegetables caused by Sclerotinia sclerotiorum.

Sclerotinia mycelium was grown in a Petri plate containing a dextrose potato Agar medium ; after 15 days the mycelium with the sclerotia was incorporated in the germination loam placed in plastic containers commonly used in the production of young plants.

After 10 days from inoculation the containers were sown with salad (Cichorium Spp). For the sowing, biological seeds, not treated with chemical products, were used.

The theses assessed were as follows : A) INOCULATED SOIL : No treatment Soil treated with 1 g/1 berberine equivalent to 1000 ppm Treated seeds + soil treated with 1 g/1 berberine equivalent to 1000 ppm Soil treated with 2 g/1 berberine equivalent to 2000 ppm Treated seeds + soil treated with 2 g/1 berberine equivalent to 2000 ppm Soil treated with 4 g/1 ROVPAL WP (Iprodion 50%) Treated seeds + soil treated with 4 g/1 ROVRAL WP B) NON-INOCULATED SOIL No treatment Seeds were immersed in the aqueous solution containing the above mentioned products for 5 minutes, and then sown on September 22, 2000.

3 repetitions per thesis were conducted.

On October 17, 2000 the effects of the treatment were assessed on germinated plants, as reported in Table 22.

Table 22 ASSESSMENT October 17, 2000 Thesis scale Disease 0 1 2 3 4 index Test with soil 3 29 32 16 20 221 inoculated Berberine lg/l 10 30 33 12 15 192 treatment in soil Berberine lg/l 17 39 31 8 5 145 treated seeds + treatment in soil Berberine 2g/1 18 52 17 8 5 130 treatment in soil Berberine 2g/1 46 32 18 3 1 81 treated seeds + treatment in soil Rovralw WP 46 20 26 7 1 97 treatment in soil Rovral 60 17 20 3-66 treated seeds + treatment in soil Natural soil 100----0 (not inoculated) Scale : 0 = healthy plant 1 = small brown necrotic lesion on the roots 2= roots partly getting brown 3= many brown roots and brown necrotic lesion on collar 4= dead plant Disease Index N. plants scale 1 X 1 + N. plants scale 2 X 2 + N. plants scale 3 X 3 + N. plants scale 4 x 4 The results clearly demonstrate that :

* Berberine is highly active against the fungus Sclerotinia sclerothiorum Its effectiveness significantly increases by increasing the dosage "At 2g/l, Berberine yields a result nearing that assessed with Rovral RHONE-POULENC, demonstrating to be a valid eco-friendly alternative in the fight against rot of cultivated plants.

Example 13 The example reports the results of the tests conducted against the fungus Phytophthora infestans on Potato.

The tests were conducted at Imola in Emilia Romagna (Italy) and the thesis concerned the following products : untreated plants, plants treated with berberine chloride, and as reference product, copper chloride (3 CuO X CaCl2 X 4 H20 ; Cu 50%) was used.

These applications were performed on a 4-row 25 m2 site, with 100 g/hl (100 ppm) berberine chloride and 400 g/hl copper chloride.

The treatments were performed on May 15, 2000 (at the first signs of illness) and on May 20, 2000, on formerly untreated plants.

The following table 23 reports the treatment calendar.

Table 23 DATE* Thesis untreated berberine chloride Reference Products 05/15/00 Berberine Copper chloride 05/20/00--------Berberine Copper chloride *month/day/year On the date of the first treatment, there were slight signs of illness in every thesis, the rate of affected leaf area was of about the 1%.

After 5 days from the second treatment on 30 randomly selected plants per each thesis the rate of affected leaf area was assessed. Each thesis was assessed for reference with an OEPP-designed colorimetric scale for assessing the effectiveness of the fungicides of the Phytosphora infestans on Potato.

In the subsequent table 24 the mean rate of affected leaf area is reported.

Table 24 Assessment May 20, 2000 Thesis % leaf surface % activity affected (mean) Untreated 10- berberine chloride 1 90 Reference product 1 90 Copper chloride Rate activity= mean of untreated plants-mean of treated plants <BR> <BR> ----------------,--------------------------------x 100 mean of untreated plants Results : a decrease of intensity of disease of 90% was assessed both in the thesis treated with reference product and in that treated with berberine. No signs of

phytotoxicity were detected.

Example 14 The example reports the results of the laboratory testing concerning berberine's effectiveness on mushrooms grown on an artificial substrate.

Within the scope of the study at issue, an analysis of the fungicidal activity against five vegetal pathogen fungi was performed. The ATCC (American Type Culture Collection) fungi employed in the tests are the following : Botrytis cinerea (n° 48339), Fusarium moniliforme (n° 36541), Magnaporte grisea (n° 64413). The following fungi were provided from SIAPA Ltd. : Alternaria sp. and Sclerotinia sclerothiorum using the test : measure of the diameter of growth on plate. The method for finding the MIC (Minimum Inhibiting Concentration of the fungal growth) was that of assaying gradually increasing doses of the substance incorporated in the cultural medium of the fungi themselves. Exact quantities of berberine-HCl (Fluka) were warm dissolved in the medium ; the dosages used started from 10 pg/ml, then increased to 100 g/ml and gradually to 500, 750, 1000, 2000 and 5000 g/ml, equivalent to 10, 100, 500, 750, 1000, 2000 and 5000 ppm, respectively. Having determined low doses to be less effective, and a stop of growth often not being attainable with higher doses, not all the dosages were tested on all fungi.

The tests comprised the following steps : Inoculating the fungus on Petri plates containing agar-enriched medium (Potato Dextrose Agar) and growing the former until attaining a logarithmic growth stage.

Transferring the fungus onto other Petri plates containing the same medium added with berberine HCl at the various dosages thereof.

* Measuring daily the mycelium diameter for 6 days.

* Assessing the growth inhibition at the 6th and last day in comparison to the reference growth.

The results, obtained as the mean of three experiments conducted in double, are reported in the following table : Table 25 Inhibition % of Berberine-Hcl on phytopathogens Doses Botrytis Sclerotinia Alternaria Fusarium Magnaporte tg/ml cinerea sclerotiorum moniliforme grisea 10 10 0. 7 1. 4 3. 2 100 29. 1 21. 4 11. 5 19. 5 500 46. 2 49. 4 38. 7 37. 3 31. 6 750 36. 3 55. 7 _ 1000 43 56------------------------ 2000 54 76. 4 90 53. 3 49 5000 100 100 100 63. 8 52. 5 Ref. : 0 0 0 0 0 The table highlights how, already at 500 ppm, the tested fungi are inhibited for a rate indicatively ranging from 30 to 50%, whereas at 2000 ppm Sclerotinia and Alternaria are reduced of the 76. 4% and 90%, respectively.

At higher doses, the total inhibition of Botrytis, Alternaria and Sclerotinia is attained.

The results of the above reported examples highlight the following : 1) berberine possesses a highly remarkable fungicidal activity, in many instances superior to that of the synthetic fungicides commonly used in agriculture ; 2) the persistence of action is prolonged, though further tests will be needed in order to assess the actual duration thereof ; 3) berberine is a wide-spectrum substance, demonstrating active against the main pathogen fungi belonging to the classes Ascomycetes, Phycomicetes,

Basidiomycetes and Deuteromycetes.

4) at the doses tested biologically active, on the tested plants no signs of phytotoxicity showed.

5) In particular, berberine demonstrated capable of replacing copper-base products, which, due to the high dosages employed and the age-long use thereof, are creating significant pollution problems to land and water-bearing strata.

In light of the above, it can be stated that berberine may be successfully used in the fight against several fungal pathologies of primary importance in agriculture.

It being a natural product, extractable from several plants as well as by synthesis, the use thereof may be provided both in the traditional and in the biological agriculture. A preferred application of the compounds of the present invention and/or of the compounds comprising them is by spraying/nebulizing. This application is particularly suitable for uses in the modern intensive agriculture. Moreover, although in the present description reference has often been made to berberine chloride it is understood that the fungicidal properties pertain to the berberine compound as such.

Berberine is quite a common compound in nature, and of age-long use by man, hence, used in lieu of the traditional synthetic fungicides, it may contribute to reduce the hazards for human health and the environment.

Within the scope of the present description, for classifying the fungi, the classification proposed by Gabriele Goidanich (Manuale di patologia vegetale 2nd reprint 1978, Edizioni Agricole, Bologna, Italy) was followed. However, other classifications based on different criteria were used, among which that of George N. Agrios in Plant Pathology (Academic Press Inc. 3rd Ed., 1988) may be mentioned. By mere way of example and not being limited by any scientific explanation, it may be mentioned that the family of the Peronosporaceae in the classification of Goidanich became the order of the

Peronosporales according to Agrios (Genera Phytophthora, Plasmopara), the family of the Taphrinaceae became the order of the Taphrinales (genus Taphrina), the family of the Erysiphaceae became the order of the Erysiphales (genera Erysiphe, Spaeroteca, Uncinula), the family of the Pseudosphaeriaceae became the order of the Pleosporales (genus Venturia), the family of the Helotiaceae became the order of the Helotiales (genera Monilia/Monilinia, Sclerotinia) and the families Mucedinaceae, Dematiaceae, Tuberculariaceae were merged into the order of the Hyphales or Moniliales (genera Alternaria, Botrytis, Cercospora, Fusarium, Magnaporte or Pyricularia).

An analogous fungicidal activity against phytopathogenic fungi is shown by the other compounds according to the present invention.