CHEMOTAXONOMY. OF MOZAMBICOFFEA. A. SANTA RAM, M. S. SREENIVASAN AND S. VISHVESHWARA. Central Coffee Research Institute.
J. Coffee Res. 12: (2) 42- 46, 1982
CHEMOTAXONOMY
OF MOZAMBICOFFEA
A. SANTA RAM, M. S. SREENIVASAN AND S. VISHVESHWARA Central Coffee Research Institute Coffee Research Station 577 117 Chikmagalur District, Kamataka. SUMMARY Chemical Moore,
investigated appropriately very closely Within while
constitution
C. kapakata
by simple placed
of four
Hirsch.,
chemical
Coftea
C. eugenioides,
C. selvetri x stands
of lour
tests.
in Mozambicoffec:
related to the genus
Mozambicoffea
species
C. racemosa
The than
Mozambicoffea and
results
C.
suggest
Erythrocoffea.
l.
in general
C
racemose
viz.
salvatrix that C.
Coffea
Swvnn .
euqenioides: et Phil.
C. eugenioides kapakata
appears
and to Mozambicoffea and
C. kapakata
form
was
is more to
be-
in particular. a
complex
apart.
INTRODUCTION Mozambicoffea, a subsection in the genus Coffea includesC. eugenioides and C. kapakata the two species which are very controversial in their systematic position (Chevalier 1947, Carvalho and Monaco 1967). Since C. eugenioides was indicated as one of theprobable ancestors of C. arabica (Narasimhaswamy and Vishveshwara 1961, 1967) it naturally attracts the attention of botanists. The object of the present investigation is to ascertain and confirm the relationshipof C. eugenioides and C. kapakata on certain chemical parameters. MATERIALS AND METHODS Fresh leaf and sapwood samples of C. eugenioides, C. recemoes, C. kapakata and C. salvatrix were collected from the blocks of Central' Coffee Research Institute for the various tests described by Gibbs. (1974) for Chemotaxonomic work. About 50g of the fresh plant material was washed, dried between the folds of clean blotting paper and macerated with 95% Et OH. This was boiled under reflux for one hour and was then cooled and filtered. The filtrate was utilized for Phenol, Flavonoid, Shinoda, Labat. Indole, Alkaloid and Salkowski tests. Methanolic extract was prepared in the same manner for Ellagic acid and Lieberman-Burchard tests. 42
43
RAM ET AL : CHEMOTAXONOMY
OF MOZAMBICOFFEA
Besides the above, fresh plant material was also used for Ehrlich (extract with 50% Et OH), Juglone (extract with chloroform in cold), leucoanthocyanin (extract with 2N HCI), Cigarette, Hotwater, Saponin Aurone, Tannin, Isenberg - Buchanan, Maule, Syringin and HCN tests. Results of all the tests are tabulated (Table 1). Affinities of pairs of species are evaluated as a function of the percentage of similarity with regard to various chemical tests by using the following formula (Sokal and Sneath 1963) and presented in table 2. Paired Affinity value (PA value)
}
No. of tests with similar reaction x
100
Total No. of tests
RESULTS AND DISCUSSION Simple phenols and allied compounds are known to be very important in systematic botany (Bate-Smith 1962, Ibrahim et al. 1962). In the present study, the extracts of all the four species gave positive result for phenol test. Further the phenol oxidase activity was ascertained in C. kapakata and C. racemosa by cigarette and hotwater tests. C. eugenioides responded slowly, while C. salvatrix did not respond to ihese tests. The importance of flavonoids in plant taxonomy has been stressed clearly by Bate - Smith (1963) and Harborne (1967). During the present investigation flavonid test gave positive reaction with all taxa studied. The development of deep cherry red colour in the extract indicated the presence of flavanones. Shinoda test confirmed the presence -of flavonoids but indicated the presence of flavanonols (= Dihydroflavonols). Occurrence of dihydroflavonols has been reported earlier by Lopes and Monaco (1979) in some species of Coffea. Aurones, however, were found to be absent in all these species. Syringyl compounds are known to form the building blocks of lignin in the angiosperms, whilst in the lower plants like pteridopyhtes and gymnosperms benzyl compounds are involved in lignification (Ibrahim, et al. 1962). Maule test indicated the presence of syringyl radicals in the wood of C. eugenioides and it was confirmed by the Syringin test. Syringin test gave a doubtfully positive reaction with the wood of C. racemosa also. However, Maule test gave a 'negative result with this species. Negative results for these two tests were obtained with the wood of C. kapakata and C. salvatrix.
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Trihydroxy phenolics like gallic acid and ellagic acid are known to be the precursors of tannins (Bate-Smith 1958). The presence of ellagic a cid in all the four species studied was ascertained from the results of ElIagic acid test and that of tannins by Tannin test. C setvetrix, however, gave a negative result for tannin test. It has also given a weak reaction for ellagic acid. Isenberg-Buchanan test indicated that these plants also possess catechol-tannins. Here again C salvatrix gave a negative result. The presence or absence of catechins is known to indicate in turn the existence or otherwise of leucoanthocyanins (Gibbs 1974) and this was found to be true during the present study in which C. eugenioides.· C. racemosa and C. kapakata were positive while C. salvatrix was negative for the Leucoanthocyanin test. Lieberman - Burchard test indicates the presence of triterpenoids or steroids when positive. This was found to be positive for all species. However, the Salkowski test confirmed the presence of steroids. Alkaloid test gave a positive result with the extracts of the four species under study. Lignan test was positive with the extracts of C. racemose, C. kapakata and C. salvatrix but was negative with that of C. eugenioides. Indole test was positive for C. racemosa and C. kapakata but was negative for C. eugenioides and C. salvatrix responded slightly to Saponin test while the remaining species responded negatively. Hydroxyquinones were found to be absent in all these species and this was confirmed by the result of Juglone test A. which indicates ·the presence of naphthoquinones or hydroxyquinones. Juglone test B confirmed the result of Flavonoid test and Shinoda test; but C. kapakata, however, gave a negative result. The presence of cournarins in these four species was indicated by the result of Jnglone test C. Result of Ehrlich test (i) was positive for C. racemosa and C. kepekata only and this was in conformity with the result of indole test. Result of Ehrlich test (ii) was uniformly negative for all these species. Ehrlich test (iii) indicated the presence of flavonoids and the absence of aurones in C. eugenioides. This test, however, was negative with other species. HCN and Labat tests were found to give negative results with all taxa studied, indicating the absence of cyanogenic glycosides and mythylene dioxy compounds in this group. From the foregoing account of the reaction patterns of the taxa of Mozambicoffea a picture of numerical relationships emerges (Table 2} and the total data support the following conclusions :
45
RAM ET AL:
Table Test
1.
CHEMOTAXONOMY
OF MOZAM81COFFEA
Chemical Properties of Mozambicoffea
C. eugenioides
C. racemose C. kapakata C. salvatrix
Phenol Test + + Cigarette test + + Hotwater test + + Maule test + Syringin test 7+ 7+ 4Lignan test Flavonoid test + + Shinoda test +y br +Y br Aurone test Indole test + Lieberman- Burchard test + + Salkowski test + + Saponin test + Isnberg-Buchanan test 7+ 7+ Tannin test ++ + Ellagic acid test ++ ++ Leucoanthocyanin test 4+ Labat test Alkaloid test + + HCN test Hydroxyqu inone test Juglone test A Juglone test B + .Juqlonatest C + + Ehrlich test C/H C/H i) -/; pr/+cr (ii) -/-/(iii) +y 7 + = doubtfully positive; pr = purple; cr = crimson. Table 2.
+
+ + +Y br
+ + +y br
+ +
+ +
Paired affinities
77.78
+ +
+ + + ++ +
+
+
+
+
+ CjH + pr/ + cr -1-
y br = yellowish brown;
C. racemosa
C. eugenioides C recemose C. kapakata
+
C/H
-/-1CI H = Cold/Hot;
of Mozambicoffea C. kapakata
74.07 92.59
C. salvatrix
62.96 74.07 70.37
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C. eugenioides shows close relationship with C. racemosa (PA 77.78), C. kapakata (PA 74.07) and C. salvatrix (PA 62.96) and thus· can be placed more appropriately in Mozambicoffea (Chevalier 1947) than in Erythrocoffea (Carvalho and Monaco 1967) with which its affinities are uniformly weak (PA with C. arabica and C congensis 70.37, unpublished) when compared to their interrelationships (PA values above 80.00, unpublished). C. kapakata also shows very close affinity to Mozambicoffea (PA values 74.07, 92.59, 70.37 with C. eugenioides C. racemosa and C. salvatrix respectively) and thus appears to conform to the position assigned to it by Hirschfeldt (cited by Chevalier 1947). Within Mozambicoffea C. euaenloides, C. racemosa and C kapakata seem to form a group while C. salvatrix stands slightly apart from, them by virtue of its reaction patterns.
LITERATURE Bate-Smith, E. C. 1958.
CITED
t.eucoanthocvanins -
in Dicotyledonous plants.
The nature and distribution
Bot. J. Linn. Soc. 55:
of tannins'
669-705
Bate-Smith, E. C. 1962. The phenolic constituents of plants and their ficance. I. Dicotyledons. Bot. J. Linn. Soc. 58: 95-173
taxonomic signi-
Bate-Smith, E. C. 1963. Usefulness of chemistry in plant taxonomy as illustrated by the flavonoid constituents. In Chemical Plant Taxonomy ed. T. Swain. London and New York Academic Press,-543 pp Carvalho, A. and L. C. Monaco. 1967. Genetic relationships of selected Cortes species. Ciencia Cultura 19 : 151-165 Chevalier, A. 1947.
Les Cafeiers du globe.
Paris, Paul Lechevalier. 356 pp
Gibbs, R. D. 1974. Chernotaxonornv of the flowering
plants.
Vol. I. Montreal.Mc-Gili
Queen's University Press, 680 pp Harborne, J. B. 1967. Comparative biochernistrv of the fl avonoids. London and New York Academic Press, 383 pp Ibrahim, R. K. G., H. N. Towers and R. D. Gibbs. 1962.
Syringic
indicators of differences between major groups of vascular
and sinapic plants.
acids as
Bot. J. Linn.
Soc. 58: 223-230 Lopes, C. R. and L. C. Monaco. 1979. genuS Coftea. J
Chemotaxonomic
studies of some species of the
Plant Crops. 7: 6-14
Naraslrnhaswarnv. R Land
S. Vishveshwara.
1961.
Report on hybrids between some-
diploid species of Coftea L. Indian Coffee 25: 104-109 Narasimhaswamy, R. L. and S. Vishveshwara , 1967. Progress report on hybrids between diploid species of Coitee L. Turrialba 17: 11-17 Sokal, R. Rand
P. H
A Sneath.
1963.
Francisco. W. H Freeman, 359 pp
Principles
of numerical taxonomy.
San.
