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The occurrence of glyceollins in plants related to Glycine max (L.) Merr.
BiochemicalSystematicsand Ecology,Vol. 17, No. 5, pp. 395-398, 1989. Printed in GreatBritain.
0305-1978189$3.00+0.00 © 1989PergamonPressplc.
The Occurrence of Glyceollins in Plants Related to Glycine max (L.) Merr. N. T. KEEN, J. L. INGHAM,* T. HYMOWITZ,t J. J. SIMS and S. MIDLAND Department of Plant Pathology, University of California, Riverside, CA 92521, U.S.A.; *Department of Food Science, University of Reading, Reading RG6 2AP, U.K.; 1Department of Agronomy, University of Illinois, Urbana, IL 61801, U.S.A.
Key Word Index--Glycine spp.; Teramnus spp.; Leguminosae; Phaseoleae; Glycininae; phytoalexins; glyceollins. Abstract--Several species from genera related to Glycine (Leguminosae-Phaseoleae) have been screened for the production of glyceollins in response to bacterial inoculation. Glyceollins are isoflavonoid (6a-hydroxypterocarpan) phytoalexins produced by Glycine spp. but not previously recognized elsewhere in the Phaseoleae. None of the 27 species examined was found to produce glyceollins except for three species of Teramnus which formed glyceollin I but no other glyceollin isomer. Accessions of Teramnus labialis, however, did not produce detectable amounts of glyceollin I.
Introduction Lackey [1, 2] divided the subtribe Glycininae of the tribe Phaseoleae into two groups, namely Glycine and Shuteria. The Glycine group is composed of about eight genera: Erninia Taub., Glycine Willd., Nogra Merr., Pseudeminia Verdc., Pseudovigna Verdc., Pueraria DC., Sinodolichos Verdc., and Teramnus P. Br. The Shuteria group also contains about eight genera: Amphicarpa Nutt., Cologania Kunth., Diphyllarium Gagnep., Dumasia DC., Mastersia Benth., Neonotonia Lackey, Shuteria W. & A., and Teyleria Backer. The economic importance of the subtribe Glycininae is primarily due to the cultivated soybean, Glycine max (L.) Merr. From a biosystematic point of view, the relationship between the genus Glycine and its allies is an enigma. The 14 currently recognised species in Glycine have somatic (2n) chromosome numbers of 38, 40, 78 and 80 [3]. The 78 and 80 chromosome plants are typical examples of polyploid species complexes. In addition, the Glycine species carrying 38 and 78 chromosomes are examples of aneuploid reduction in the evolution of the genus [4]. The other genera, except Terarnnus, in the Glycine group have 22 somatic chromosomes. Terarnnus have a somatic chromosome number of 28. The
(Received 6 April 1989)
genera in the Shuteria group have 2n chromosome numbers of 20, 22 and 44 [5]. Live seed collections, as well as extensive herbarium specimens, of many genera allied to Glycine are sparse to non-existent. Chromosome counts of one or two accessions per species per genus are not satisfactory in establishing generic relationships. In the absence of extensive germplasm collections of these genera, it therefore seemed logical to pursue a complementary course to determine the possible affinities of genera to Glycine. Keen et al. [6] reported that 61 accessions representing 10 species of Glycine produced one or more isomers, or close structural relatives, of the isoflavonoid (pterocarpan) phytoalexins commonly called glyceollins. Unlike other legume phytoalexins, which often occur widely within and between genera and/or subtribes [7], the glyceollins have thus far only been detected in Glycine (tribe Phaseoleae) and, in the case of glyceollin I (Fig. 1), more recently in members (e.g. Psoralea spp.) of the distantly related tribe Psoraleae (Ingham, unpublished data). Two glyceollin isomers have also been reported from the non-legume Costus speciosus [8]. Since the production of glyceollin isomers appears to be tightly conserved within Glycine spp. [6, 7], the formation of these compounds may provide a chemotaxonomic criterion for recognizing closely allied Leguminosae. For this reason, we
395
396
N.T. KEEN, J. L. INGHAM, T. HYMOWITZ, J. J. SIMS AND S. MIDLAND
OH
OH FIG. 1. GLYCEOLLIN I.
have screened accessions from various legume genera thought to be related to Glycine, as well as certain other legume species, for their production of glyceollins or related compounds. We report here that none of them, except for certain accessions of Teramnus, produce glyceollins.
Results and Discussion As shown in Table 1, most of the tested plants failed to make detectable glyceollin isomers in response to leaflet inoculation with the bacterium, Pseudomonas syringae pv. pisi, a non-pathogen of the tested plants. Similar results were also obtained when the leaflets were tested (drop-diffusate method [7]) with a conidial suspension of the fungus, Helminthosporium carbonum (Ingham, unpublished data). However, confirming previous observations [7], our accessions of Pueraria Iobata formed tuberosin [9], a glyceollin-like pterocarpan, but none yielded detectable quantities of any glyceollin isomer. Other species, e.g. Cajanus albicans (cajanol and cajanin) and Neonotonia wight/)" (genistein and wighteone) afforded known compounds previously obtained from these or related legumes [10-12], following inoculation with either H. carbonum [10, 11] or Phytophthora megasperma var. sojae [12]. Of the species listed in Table 1, only seven accessions of Teramnus representing three species (repens, micans, and uncitatus), as well as one unidentified Teramnus accession, made glyceollin I (Fig. 1) but no other detectable glyceollin isomer in response to bacterial inoculation. In contrast, five different accessions of a fourth Teramnus species (T. labia/is) were apparently unable to produce glyceollins. Instead, these accessions accumulated an antifungal compound which, based on preliminary spectroscopic and chromatographic data, appears to be a benzofuran-type phytoalexin, related to, or identical
to, either vignafuran [13] or its 6-demethyl analogue [14]. Although initial screening of T. labia/is CU232 revealed the formation of glyceollin I, it was subsequently suspected that the seed sample was in fact a mixture of Teramnus species. Further experiments with authenticated seed of T. labia/is (accessions CU211, 221, 222, 232 and 233) uniformly failed to disclose the production of any glyceollin isomer. The surprising absence of glyceollins in related legume plants is most readily interpreted as indicating that genes encoding specific prenyl transferases and the corresponding cyclizing enzymes are not widely distributed in the group. For example, Zahringer et aL [18] demonstrated that G. max contains two specific transferases catalysing prenylation of the hydroxy pterocarpan, glycinol, specifically at either the 2 or 4 positions. Neither enzyme, however, caused prenylation at any other aromatic carbon besides its unique site. While biogenetic evidence clearly suggests that analogous 2 and 4 prenyl transferases are widely distributed in legumes, either the 6a hydroxy pterocarpan substrate or required prenyl cyclizing enzymes must not generally be present as in G. max. Of these, the most unique enzyme is the 6a hydroxylase, recently characterized by Hagmann et aL [19] and shown to utilize molecular oxygen by Matthews et al. [20].
Experimental A total of 42 legume accessions were used in the present study, representing members of the subtribe Glycininae, as well as an assortment of species from other subtribes of the Phaseoleae (Table 1). Voucher specimens of all the accessions have been deposited in the herbarium of the Crop Evolution Laboratory, University of Illinois at Urbana. The various accessions shown in Table 1 were grown from seed, and trifoliate leaves inoculated with Pseudomonas syr/ngae pv. pisi cells as previously described [6J. In most cases, this treatment resulted in a visible hypersensitive defence reaction of the inoculated area within 48 h. In many plants, this reaction is known to result in the accumulation of phytoalexins. Inoculated leaves were removed from the plants after 3 days and phytoalexins extracted and chromatographed on silica gel TLC plates (GF-254), as previously outlined [6]. Glyceollin was detected as a fluorescence-quenching spot, corresponding to an authentic standard of glyceollin I (Fig. 1), when the chromatograms were inspected under short wavelength (254 nm) UV light. Compounds were subsequently eluted from the silica gel with acetone. No attempts were made to quantify the glyceollin thus obtained. Several species also yielded spots distinct from the glyceollin marker, and these compounds were in some cases recovered for analysis. Glyceollin samples from TLC plates were further purified by HPLC according to the
GLYCEOLLINS IN PLANTS RELATED TO GLYClNE M A X (L.) MERR.
397
TABLE 1. PLANTS SCREENED FOR GLYCEOLLIN PRODUCTION
Accession1" * Amphicarpa bracteata (L.) Fernald Amphicarpa bracteata * Amphicarpa edgeworthii Benth. Cajanus albicans (W.+A.) van der Maesen Cajanus cajan (L.) Millsp.II Cajanus scarabaeoides (L.) Thourss Cyamopsis tetragonoloba (L.) Taub¶ * Dumasia villosa DC. Galactia Iongifolia (Jacq.) Benth * Giycine max (L.) Merr. cv. Harosoy Macroptilium atropurpureum (DC.) Urb. * Neonotonia wightii (Arnott) Lackey * Neonotonia wight/)" (Arnott) Lackey * Pueraria Iobata (Willd.) Ohwi * * * *
Pueraria Pueraria Pueraria Pueraria
Iobata (Willd.) Ohwi Iobata (Willd.) Ohwi Iobata (Willd.) Ohwi phaseoloides (Rox.) Benth.
* Pueraria phaseoloides (Rox.) Benth. * Pueraria phaseoloides (Rox.) Benth. Pseudoeriosema boriani (Schweinf.) Hauman * Pseudeminia comosa (Bak.) Verdc. Pseudeminia comosa (Bak.) Verdc. * Pseudeminia sp. * Pseudovigna argentea (Willd.) Verd. Sphenostylis stenocarpa (A. Rich) Harms Strophostyles helvola (L.) Ell. Strophostyles helvola (L.) Ell. * Teramnus sp. * Teramnus labialis (L.) Spreng. * * * * * * * * * * * *
(L.) Spreng. (L.) Spreng. (L.) Spreng. (L.) Spreng. Teramnus repens (Taub.) Bak. f. Teramnus m/cans (Bak.) Bak, Teramnus uncitatus (L.) Sw. Teramnus uncitatus (L.) Sw. Teramnus uncitatus (L.) Sw. Teramnus uncitatus (L,) Sw. Teramnus uncitatus (L.) Sw. Teyleria koordersii (Backer) Bacher
Teramnus Teramnus Teramnus Teramnus
labia/is labia/is labia/is labia/is
CU no.
Glyceollin$ isomer
Other4:§ compounds
169 200 243 193 188 196 168 293 284
_
m
m
--
Unknown
--
Cajanol, Cajanin
compound
--
Unknown compound
--
Unknown
compound
I, II, III,
186 145 146 36 37 38 4 3 6 29 152 5 11 26 283 14 166 173 199 211 221 222 232 233 220 164 223 227 163 228 253 42
--
-----
--
Genistein, Wighteone Genistein, Wighteone Tuberosin Tuberosin Tuberosin Tuberosin
--
Unknown
compound
--
Unknown
compound
--
Unknown compound
--
Unknown
compound
--
Unknown
compound
--
Unknown
compound
--
Unknown
compound
I
----
Unknown compound** Unknown compound** Unknown compound**
--
Unknown
--
Unknown compound**
compound**
*Plants classified within the subtribe Glycininae. t A collection of legumes maintained at the University of Illinois, Urbana; CU numbers are those of the Champaign-Urbane seed collection. ~Glyceolin isomers denoted were isolated by HPLC and identified by comparison with authentic compounds; some accessions yielded compounds which were not identified, others yielded known compounds, and other plants did not produce significant quantitites of phytoalexin as denoted by (--). §A previous study of phytoalexin production by leaflets of Shuteria vestita (subtribe Glycininae) has revealed three antifungal flavanones but no evidence for the production of glyceollins or other isoflavonoid compounds [15]. IIKnown to produce both cajanin and cajsnol after fungal inoculation of etiolatsd stems [10, 11]. ITLeguminosae-Papilionoidea, tribe Indigofereae. **Probably a benzofuran-type compound (see text).
398
N. 1. KEEN,J. L. INGHAM,T. HYMOWITZ, J. J. SIMS AND S. MIDLAND
method outlined in ref. [6] which allows separation of isomers I (Fig. 1), II and II1. The identity of glyceollin I (and also glyceollins II and III in the case of G. max) was confirmed by comparison of spectroscopic (UV, MS, 1NMR) data with previously published values [16, 17].
8. Kumar, S., Shukla, R. S., Singh, K. P., Paxton, J. D., and Husain, A. (1984) Phytopathology74, 1349. 9. Joshi, B. S. and Kumat, V. N. (1973) J. Chem. Soc. Perkin Trans. I, 907. 10. Ingham, J. L. (1976) Z. Natufforsch. 31¢, 504. 11. Ingham, J. L. (1979) Z. Naturforsch. 34c, 159. 12. Ingham, J. L., Keen, N. T. and Hymowitz, T. (1977) Phytochemistry 16, 1943. 13. Preston, N. W., Chamberlain, K. and Skipp, R. A. (1975) Phytochemistry 14, 1843. 14. Ingham, J. L. and Dewick, P. M. (1978) Phytochemistry 17, 535. 15. Ingham, J. L., Tahara, S. and Dziedzic, S. Z. (1986) J. NaL Prod. (Lloydia) 49, 631. 16. Burden, R. S. and Bailey, J. A. (1975) Phytochemistry 14, 1389. 17. Lyne, R., Mulheirn, L. F. and Leworthy, D. P. (1976) J. Chem. Soc. Chem. Commun., 497. 18. Z~ihringer, U., Schaller, E. and Grisebach, H. (1981) Z. Naturforach. 36c, 234. 19. Hagmann, M. L., Heller, W. and Grisebach, H. (1984) Eur. J. B/ochem. 142, 127. 20. Matthews, D. E., Plattner, R. D. and VanEtten, H. D. (1989) Phytochemistry 28, 113.
References 1. Lackey, J. A. (1977) A Synopsis ofPhaseo/eae (Leguminosae, Papi//onoideae). Ph.D. Dissertation, Iowa State University, Ames, Iowa. 2. Lackey, J. A. (1981) in Advances in Legume SystemaUcs, Part 1 (Polhill, R. M. and Raven, P. H., eds), pp. 301-327. Royal Botanic Gardens, Kew. 3. Singh, R. J., Kollipara, K. P. and Hymowitz, T. (1988) Genome 30, 166. 4. Singh, R. J., KoUipara, K. P. and Hymowitz, 1". (1987) Genome 29, 490. 5. Kumar, P. S. and Hymowitz, T. (1989) Euphytica (in press). 6. Keen, N. T., Lyne, R. L., and Hymowitz, T. (1986) Biochem. SysL Ecol. 14, 481. 7. Ingham, J. L. (1982) in Phytoa/ex/ns (Bailey, J. A. and Mansfield, J. W., eds), pp. 21-80, Wiley & Sons, New York.
0305-1978189$3.00+0.00 © 1989PergamonPressplc.
The Occurrence of Glyceollins in Plants Related to Glycine max (L.) Merr. N. T. KEEN, J. L. INGHAM,* T. HYMOWITZ,t J. J. SIMS and S. MIDLAND Department of Plant Pathology, University of California, Riverside, CA 92521, U.S.A.; *Department of Food Science, University of Reading, Reading RG6 2AP, U.K.; 1Department of Agronomy, University of Illinois, Urbana, IL 61801, U.S.A.
Key Word Index--Glycine spp.; Teramnus spp.; Leguminosae; Phaseoleae; Glycininae; phytoalexins; glyceollins. Abstract--Several species from genera related to Glycine (Leguminosae-Phaseoleae) have been screened for the production of glyceollins in response to bacterial inoculation. Glyceollins are isoflavonoid (6a-hydroxypterocarpan) phytoalexins produced by Glycine spp. but not previously recognized elsewhere in the Phaseoleae. None of the 27 species examined was found to produce glyceollins except for three species of Teramnus which formed glyceollin I but no other glyceollin isomer. Accessions of Teramnus labialis, however, did not produce detectable amounts of glyceollin I.
Introduction Lackey [1, 2] divided the subtribe Glycininae of the tribe Phaseoleae into two groups, namely Glycine and Shuteria. The Glycine group is composed of about eight genera: Erninia Taub., Glycine Willd., Nogra Merr., Pseudeminia Verdc., Pseudovigna Verdc., Pueraria DC., Sinodolichos Verdc., and Teramnus P. Br. The Shuteria group also contains about eight genera: Amphicarpa Nutt., Cologania Kunth., Diphyllarium Gagnep., Dumasia DC., Mastersia Benth., Neonotonia Lackey, Shuteria W. & A., and Teyleria Backer. The economic importance of the subtribe Glycininae is primarily due to the cultivated soybean, Glycine max (L.) Merr. From a biosystematic point of view, the relationship between the genus Glycine and its allies is an enigma. The 14 currently recognised species in Glycine have somatic (2n) chromosome numbers of 38, 40, 78 and 80 [3]. The 78 and 80 chromosome plants are typical examples of polyploid species complexes. In addition, the Glycine species carrying 38 and 78 chromosomes are examples of aneuploid reduction in the evolution of the genus [4]. The other genera, except Terarnnus, in the Glycine group have 22 somatic chromosomes. Terarnnus have a somatic chromosome number of 28. The
(Received 6 April 1989)
genera in the Shuteria group have 2n chromosome numbers of 20, 22 and 44 [5]. Live seed collections, as well as extensive herbarium specimens, of many genera allied to Glycine are sparse to non-existent. Chromosome counts of one or two accessions per species per genus are not satisfactory in establishing generic relationships. In the absence of extensive germplasm collections of these genera, it therefore seemed logical to pursue a complementary course to determine the possible affinities of genera to Glycine. Keen et al. [6] reported that 61 accessions representing 10 species of Glycine produced one or more isomers, or close structural relatives, of the isoflavonoid (pterocarpan) phytoalexins commonly called glyceollins. Unlike other legume phytoalexins, which often occur widely within and between genera and/or subtribes [7], the glyceollins have thus far only been detected in Glycine (tribe Phaseoleae) and, in the case of glyceollin I (Fig. 1), more recently in members (e.g. Psoralea spp.) of the distantly related tribe Psoraleae (Ingham, unpublished data). Two glyceollin isomers have also been reported from the non-legume Costus speciosus [8]. Since the production of glyceollin isomers appears to be tightly conserved within Glycine spp. [6, 7], the formation of these compounds may provide a chemotaxonomic criterion for recognizing closely allied Leguminosae. For this reason, we
395
396
N.T. KEEN, J. L. INGHAM, T. HYMOWITZ, J. J. SIMS AND S. MIDLAND
OH
OH FIG. 1. GLYCEOLLIN I.
have screened accessions from various legume genera thought to be related to Glycine, as well as certain other legume species, for their production of glyceollins or related compounds. We report here that none of them, except for certain accessions of Teramnus, produce glyceollins.
Results and Discussion As shown in Table 1, most of the tested plants failed to make detectable glyceollin isomers in response to leaflet inoculation with the bacterium, Pseudomonas syringae pv. pisi, a non-pathogen of the tested plants. Similar results were also obtained when the leaflets were tested (drop-diffusate method [7]) with a conidial suspension of the fungus, Helminthosporium carbonum (Ingham, unpublished data). However, confirming previous observations [7], our accessions of Pueraria Iobata formed tuberosin [9], a glyceollin-like pterocarpan, but none yielded detectable quantities of any glyceollin isomer. Other species, e.g. Cajanus albicans (cajanol and cajanin) and Neonotonia wight/)" (genistein and wighteone) afforded known compounds previously obtained from these or related legumes [10-12], following inoculation with either H. carbonum [10, 11] or Phytophthora megasperma var. sojae [12]. Of the species listed in Table 1, only seven accessions of Teramnus representing three species (repens, micans, and uncitatus), as well as one unidentified Teramnus accession, made glyceollin I (Fig. 1) but no other detectable glyceollin isomer in response to bacterial inoculation. In contrast, five different accessions of a fourth Teramnus species (T. labia/is) were apparently unable to produce glyceollins. Instead, these accessions accumulated an antifungal compound which, based on preliminary spectroscopic and chromatographic data, appears to be a benzofuran-type phytoalexin, related to, or identical
to, either vignafuran [13] or its 6-demethyl analogue [14]. Although initial screening of T. labia/is CU232 revealed the formation of glyceollin I, it was subsequently suspected that the seed sample was in fact a mixture of Teramnus species. Further experiments with authenticated seed of T. labia/is (accessions CU211, 221, 222, 232 and 233) uniformly failed to disclose the production of any glyceollin isomer. The surprising absence of glyceollins in related legume plants is most readily interpreted as indicating that genes encoding specific prenyl transferases and the corresponding cyclizing enzymes are not widely distributed in the group. For example, Zahringer et aL [18] demonstrated that G. max contains two specific transferases catalysing prenylation of the hydroxy pterocarpan, glycinol, specifically at either the 2 or 4 positions. Neither enzyme, however, caused prenylation at any other aromatic carbon besides its unique site. While biogenetic evidence clearly suggests that analogous 2 and 4 prenyl transferases are widely distributed in legumes, either the 6a hydroxy pterocarpan substrate or required prenyl cyclizing enzymes must not generally be present as in G. max. Of these, the most unique enzyme is the 6a hydroxylase, recently characterized by Hagmann et aL [19] and shown to utilize molecular oxygen by Matthews et al. [20].
Experimental A total of 42 legume accessions were used in the present study, representing members of the subtribe Glycininae, as well as an assortment of species from other subtribes of the Phaseoleae (Table 1). Voucher specimens of all the accessions have been deposited in the herbarium of the Crop Evolution Laboratory, University of Illinois at Urbana. The various accessions shown in Table 1 were grown from seed, and trifoliate leaves inoculated with Pseudomonas syr/ngae pv. pisi cells as previously described [6J. In most cases, this treatment resulted in a visible hypersensitive defence reaction of the inoculated area within 48 h. In many plants, this reaction is known to result in the accumulation of phytoalexins. Inoculated leaves were removed from the plants after 3 days and phytoalexins extracted and chromatographed on silica gel TLC plates (GF-254), as previously outlined [6]. Glyceollin was detected as a fluorescence-quenching spot, corresponding to an authentic standard of glyceollin I (Fig. 1), when the chromatograms were inspected under short wavelength (254 nm) UV light. Compounds were subsequently eluted from the silica gel with acetone. No attempts were made to quantify the glyceollin thus obtained. Several species also yielded spots distinct from the glyceollin marker, and these compounds were in some cases recovered for analysis. Glyceollin samples from TLC plates were further purified by HPLC according to the
GLYCEOLLINS IN PLANTS RELATED TO GLYClNE M A X (L.) MERR.
397
TABLE 1. PLANTS SCREENED FOR GLYCEOLLIN PRODUCTION
Accession1" * Amphicarpa bracteata (L.) Fernald Amphicarpa bracteata * Amphicarpa edgeworthii Benth. Cajanus albicans (W.+A.) van der Maesen Cajanus cajan (L.) Millsp.II Cajanus scarabaeoides (L.) Thourss Cyamopsis tetragonoloba (L.) Taub¶ * Dumasia villosa DC. Galactia Iongifolia (Jacq.) Benth * Giycine max (L.) Merr. cv. Harosoy Macroptilium atropurpureum (DC.) Urb. * Neonotonia wightii (Arnott) Lackey * Neonotonia wight/)" (Arnott) Lackey * Pueraria Iobata (Willd.) Ohwi * * * *
Pueraria Pueraria Pueraria Pueraria
Iobata (Willd.) Ohwi Iobata (Willd.) Ohwi Iobata (Willd.) Ohwi phaseoloides (Rox.) Benth.
* Pueraria phaseoloides (Rox.) Benth. * Pueraria phaseoloides (Rox.) Benth. Pseudoeriosema boriani (Schweinf.) Hauman * Pseudeminia comosa (Bak.) Verdc. Pseudeminia comosa (Bak.) Verdc. * Pseudeminia sp. * Pseudovigna argentea (Willd.) Verd. Sphenostylis stenocarpa (A. Rich) Harms Strophostyles helvola (L.) Ell. Strophostyles helvola (L.) Ell. * Teramnus sp. * Teramnus labialis (L.) Spreng. * * * * * * * * * * * *
(L.) Spreng. (L.) Spreng. (L.) Spreng. (L.) Spreng. Teramnus repens (Taub.) Bak. f. Teramnus m/cans (Bak.) Bak, Teramnus uncitatus (L.) Sw. Teramnus uncitatus (L.) Sw. Teramnus uncitatus (L.) Sw. Teramnus uncitatus (L,) Sw. Teramnus uncitatus (L.) Sw. Teyleria koordersii (Backer) Bacher
Teramnus Teramnus Teramnus Teramnus
labia/is labia/is labia/is labia/is
CU no.
Glyceollin$ isomer
Other4:§ compounds
169 200 243 193 188 196 168 293 284
_
m
m
--
Unknown
--
Cajanol, Cajanin
compound
--
Unknown compound
--
Unknown
compound
I, II, III,
186 145 146 36 37 38 4 3 6 29 152 5 11 26 283 14 166 173 199 211 221 222 232 233 220 164 223 227 163 228 253 42
--
-----
--
Genistein, Wighteone Genistein, Wighteone Tuberosin Tuberosin Tuberosin Tuberosin
--
Unknown
compound
--
Unknown
compound
--
Unknown compound
--
Unknown
compound
--
Unknown
compound
--
Unknown
compound
--
Unknown
compound
I
----
Unknown compound** Unknown compound** Unknown compound**
--
Unknown
--
Unknown compound**
compound**
*Plants classified within the subtribe Glycininae. t A collection of legumes maintained at the University of Illinois, Urbana; CU numbers are those of the Champaign-Urbane seed collection. ~Glyceolin isomers denoted were isolated by HPLC and identified by comparison with authentic compounds; some accessions yielded compounds which were not identified, others yielded known compounds, and other plants did not produce significant quantitites of phytoalexin as denoted by (--). §A previous study of phytoalexin production by leaflets of Shuteria vestita (subtribe Glycininae) has revealed three antifungal flavanones but no evidence for the production of glyceollins or other isoflavonoid compounds [15]. IIKnown to produce both cajanin and cajsnol after fungal inoculation of etiolatsd stems [10, 11]. ITLeguminosae-Papilionoidea, tribe Indigofereae. **Probably a benzofuran-type compound (see text).
398
N. 1. KEEN,J. L. INGHAM,T. HYMOWITZ, J. J. SIMS AND S. MIDLAND
method outlined in ref. [6] which allows separation of isomers I (Fig. 1), II and II1. The identity of glyceollin I (and also glyceollins II and III in the case of G. max) was confirmed by comparison of spectroscopic (UV, MS, 1NMR) data with previously published values [16, 17].
8. Kumar, S., Shukla, R. S., Singh, K. P., Paxton, J. D., and Husain, A. (1984) Phytopathology74, 1349. 9. Joshi, B. S. and Kumat, V. N. (1973) J. Chem. Soc. Perkin Trans. I, 907. 10. Ingham, J. L. (1976) Z. Natufforsch. 31¢, 504. 11. Ingham, J. L. (1979) Z. Naturforsch. 34c, 159. 12. Ingham, J. L., Keen, N. T. and Hymowitz, T. (1977) Phytochemistry 16, 1943. 13. Preston, N. W., Chamberlain, K. and Skipp, R. A. (1975) Phytochemistry 14, 1843. 14. Ingham, J. L. and Dewick, P. M. (1978) Phytochemistry 17, 535. 15. Ingham, J. L., Tahara, S. and Dziedzic, S. Z. (1986) J. NaL Prod. (Lloydia) 49, 631. 16. Burden, R. S. and Bailey, J. A. (1975) Phytochemistry 14, 1389. 17. Lyne, R., Mulheirn, L. F. and Leworthy, D. P. (1976) J. Chem. Soc. Chem. Commun., 497. 18. Z~ihringer, U., Schaller, E. and Grisebach, H. (1981) Z. Naturforach. 36c, 234. 19. Hagmann, M. L., Heller, W. and Grisebach, H. (1984) Eur. J. B/ochem. 142, 127. 20. Matthews, D. E., Plattner, R. D. and VanEtten, H. D. (1989) Phytochemistry 28, 113.
References 1. Lackey, J. A. (1977) A Synopsis ofPhaseo/eae (Leguminosae, Papi//onoideae). Ph.D. Dissertation, Iowa State University, Ames, Iowa. 2. Lackey, J. A. (1981) in Advances in Legume SystemaUcs, Part 1 (Polhill, R. M. and Raven, P. H., eds), pp. 301-327. Royal Botanic Gardens, Kew. 3. Singh, R. J., Kollipara, K. P. and Hymowitz, T. (1988) Genome 30, 166. 4. Singh, R. J., KoUipara, K. P. and Hymowitz, 1". (1987) Genome 29, 490. 5. Kumar, P. S. and Hymowitz, T. (1989) Euphytica (in press). 6. Keen, N. T., Lyne, R. L., and Hymowitz, T. (1986) Biochem. SysL Ecol. 14, 481. 7. Ingham, J. L. (1982) in Phytoa/ex/ns (Bailey, J. A. and Mansfield, J. W., eds), pp. 21-80, Wiley & Sons, New York.