30. Dillenia suffruticosa (Griff. 1854) Martelli 1886

Wormia suffruticosa G-riff., Not. 4, 1854, p. 706 & *Ic. PI. As., 1854, t. 649; Hook.f. & Thorns., Fl. Br. Ind. 1, 1872, p. 35; Villar, Nov. App., 1880, p. 347; King, J. As. Soe. Beng. 58, II, 1889, p. 364; Ridl, J. Str. Br. R. A. S. 54, 1910, p. 5; Back., Schoolfl. Java, 1911, p. 10; Koord., Exk. Fl. Java 2, 1912, p. 601; "Blaauw, Trop. nat. in sc.hotsen on kleuren, 1913, p. 17, t. 12; Rid]., Fl. Mai. Pen. 1, 1922, p. 8; Burk., Diet. Eeon. Prod. Mai. Pen., 1935, p. 2265; Corn., Gard. Bull. S. S. 10, 1939, p. 9; 'Corn., Wayside Trees Malaya, 1940, p. 207, pi. 53; Back., Bekn. Fl. Java (em. ed.) 4, 1942, fana. 80, p. 4.

Wormia excelsa Auct. non Jack; Hook.f. & Thorns., Fl. Ind. 1, 1855, p. 67. Wormia subsessilis Miq., Fl. Ind. Bat. Suppl., 1860, p. 619; Teysm. & Binn., J. Bot. Nderl. 1, 1861, p. 364; "Miq., Ann. Mus. Bot. Lugd. Bat. 1,"l864, p. 315, t. 9; Miq., Ann. Mus. Bot. Lugd. Bat. 4, 1868, p. 77; Ridl., J. Str. Br. R. A. S. 54, 1910, p. 4; Ridl., Saraw. Mus. .T. 1, 1913, p. 71; 'Ridl., Fl. Mai. Pen. 1, 1922, p. 7.

Wormia revoluta Teysm. Sc Binn., J. Bot. Neerl. 1, 1861, p. 364, in syn.

Wormia burbidgei '"Hook.f., Bot. Mag., 1880, t. 6531.

Dillenia suffruticosa (Griff.) Martelli in Becc., Malesia 3, 1886, p. 163; Fin. & Gagnep., Bull. Soc. Bot. Fr. Mem. 4, 1906, p. 10; Merr., Bibl. En. Bom. PI., 1921, p. 384; de Wit, Bull. Bot, Gard. Btzg III, 18, 1949, p. 208; 'Hoogl., Fl. Mai. I, 4, 1951, p. 162.

Dillenia burbidgei (Hook.f.) Grig in Engl. & Prantl, Nat. Pfl. Fam. 3, 6, 1893, p. 123.

Dillenia suffruticosa var. borneensis (Ridl.) Ridl., Saraw. Mus. J. 1, 1913, p. 71; Merr., Bibl. En. Born. PI., 1921, p. 384.

Type specimens: Wormia suffruticosa: Griffith Kew Distr. 55, Malacca, 1845; holotype in K, isotvpes in C, OGE, PI, GH, K, L, M, NY, P. — Wormia subsessilis: Tcysmann 3203 HE, Banka; liolotypo in TJ, isotypes in BZ, CAL, K, L, MEL. — Wormia burbidgei: Burbidge, Borneo, 1877—8; holotype in K. — Wormia subsessilis var. borneensis: Hewitt, Kuching, 1909; holotype in SING, isotypo in K.

Large shrubs or small trees, evergreen, up to 10 m high, with stout, brown trunk, often forming thickets. Branches sympodial, younger ones ca 3 mm thick, glabrous to densely villose with 2—3 nun long hairs, more or less glabrescent. Leaf-scars amplexicaul, for about 1 /2 single line, for Y, subfalcate with ca 20 leaf-traces near lower margin. Leaves elliptic to" ovate, (10—) 15—25(—45) X (5—)8—12(—26) cm, with (7—)12—20 (—27) nerves on either side ; rounded to obtuse at apex, obtuse at base, decurring into petiolar wings; margin entire to dentate or doubly dentate, nerves ending in apex of teeth, in larger leaves often 1—3 rather strong secondary nerves directed downward near margin, ending in apex of Smaller teeth; glabrous, rarely slightly villose on intervenium in younger leaves only above, slightly to densely villose on nerves, on both sides along midrib (continuing on petiole), and along line which delimits budenclosing part of leaf-base beneath. Petiole 2—6 cm, with amplexicaul wings; wings 4—40(—15) mm broad, nervation of blade continuing, birt

less marked, on wings, wings and base of blade below line where both sides cohere in young leaf of different colour (darker when dry) ; wings usually persistent. Inflorescences terminal, (4 —)6—10 (—18)-flowered, up to 30 cm long, simple racemes or composed by having lateral branch at place of second, sometimes also third flower; axis ca 3 mm thick, glabrous to, particularly when young, densely villose; bracts caducous, triangular, 6—15 X 3—5 mm. Flowers 8—12 cm across. Pedicel 0.8—3 cm long, 2— 3 mm thick, thickened to 3—4 mm at apex, without bracteoles. Sepals 5, obovate, 15—22 X 8—12 mm, glabrous inside, glabrous to rather densely villose outside. Petals 5, bright yellow, obovate, 40—50 X 25—30 mm, rounded at apex, narrowed towards base. Androecium with distinct group of staminodes on outer side. Staminodes ca 100, linear, ca 4—6 X 0.3 mm, yellow, obtuse at apex. Stamens ca 175, outermost ones slightly curved in bud, ca 8 mm long, innermost ones with apical part reflexed outward in bud, ca 13 mm long, with stamens of intermediate lengths between; filament of outermost stamens ca 3 1 / 2, of innermost ones ca 2 mm long; anther ca 0.5 mm broad, obtuse at apex; thecae linear, opening with pore near apex on outer side. Carpels 5—8, usually 7, arranged around sharp conical receptacle, light green, elliptic, ca 5 X 2 mm, glabrous, each with 7—10 ovules; styles spreading, filamentous, ca 10 mm long, 0.5 mm thick, yellowish white. Pseudocarps dehiscent, when ripe sepals enlarged to 18—25 X 10—15 mm; carpels red, 20—25 X 10—16 mm, each with 1—4 seeds. Seeds obovoid, ca 3 X 2 mm, brown or black, enclosed by scarlet, membranous aril.

Sumatra: Palembang: Pladju, Polak 140, fl. & fr. Oct. 1030 (BZ). — Banka: H ors field, fl. (BM); Kurz 425, fl. (K) ; Baturasak, Amand, fl. Juno 1858 (U) ; Ploem, fl. & fr. (L) ; Djcbus, Berlchowt 7g, fl. Aug. 1886 (BZ) ; Berkhout (BZ) ; G. Menumbing, near Muntok, Biinnemeyer 1S51, fl. & fr. Oct. 1917 (BZ, L) ; Bakem, Sungai Liât, Biinnemeyer 1731, fl. Oct. 1917 (BZ) ; Sungai Lajang, Sungai Liât, Biinnemeyer 1807, fl. & fr. Oct. 1917 (BZ, L) ; Sungai Liât, Rcbo, Biinncmeyer 2510, fl. Nov. 19.17 (B'Z) ; near Pangkalpinang-Belinju, Huitema 24, fl. 1932 (IIZ)'; Bakit, Belinju, Coert 1630, fl. Sept. 1941 (L). — Billiton: Riedel, fl. (FI); S of Manggar,, n<wi 32, fr. March 1907 (BZ). — Riouw-Lingga Arch.: Pulau Tudjuh, Ajer Suar, Biinnemeyer 5966, fl. & fr. May 1919 (BZ, L) ; Sungai Tanda, Pulau Liugga, Biinnemeyer 6933, fl. July 1919 (BZ).

Malay Peninsula: Walker 224, fr. (GL, P) ; Suugai Iwong, Goodenough, fl. May 1892 (RM). — Pcrak: Maxwell's Hill, Taiping, Henderson SF 10014, fl. & fr. 1922 (SING) ; Taiping, Henderson SF 10358, fr. Jan. 1923 (SING) ; Tinmines, Taiping, llaniff SF 13130, March 1924 (SING). — Pahang: Balok, Teop CF 3616, fr. Jan. 1920 (K, SING). — Selangor: Rawang Bistr., Goodenough 10470, fl. March 1809 (SING) ; Garcona Kuala Lumpur, Kalong CF 17479, fl. Aug. 1929 (SING); Fran ok 1115, fl. & fr. Sept. 1937 (C) ; Tinmines, Ranching, Nv.r SF 34430, fl. & fr. Nov. 1937 (MO, SING). — Negri Sembilan: Seremban, -A• Ivins 1783, fr. July 1885 (SING). - Malacca: fr. (SING) ; Gmidichaud 26, fl. & fr. Feb. 18137 (G, P) ; o _o_. i r> A C\ /n \ . A Ti N ... Cuming 2858, fl. 1840 (BM, CÖE, K) ; Delesscrt, fl. & fr. 1840 (G) ; Ayer Panas, x , , , ... v , , . , Griffith (Herb. E. I. Comp. 55) , fl. & fr. 184-5 (C, CGE, PI, GH, IC, L, M, NY, P) ; Griffith, fr. (OAL) ; Maingay 1020 (Kcw Dixtr.■. 4), fl. 1865—6 (K) ; Maingay lOSOA (Kew Distr. 4), fl. May 1868 (GAIL, K, L) ; A Ivins, fl. & fr. (SING); Bukit Braang, Uolmberg 710, fl. & fr. Apr. 1891 (MEL, SING); Ilervey, fl. 1S91 (BM, GAL, SING) ; Ilervey, fl. & fr. Apr. 1893 (CAL, P) ; Ilervey, fr. (A). — Joliore: Pinyerong, Ridley, fr. May 1889 (SING); Pulau Tekong, Eidley 3966, fl. 1890 (SING); Eidley, fl. 1890 (BM) ; Kuala Sedili Besar, Feilding, fl. 1892 (SING); Sedili Kechik, Yeob CF 5832, fl. & fr. July 1921 (SING) ; Mawai, Sedili, Corner SF 21189, fl. & fr. May 1932 (K, NY, SING) ; Sungai Sedili, Corner SF 32984, fl. & fr. May 1937 (L, SING) ; Sungai Sedili, near Mawai, Corner SF 33546 $• 33547, fl. & fr. June 1937 (L, SING). — Pulau Penan g : Delessert, fl. & fr. 1835 (G). — Singapore: Thomson 10, fl. (K, P) ; Lobb 329, fl. (OGiE, FI, G, GH, GL, K, MEL, OX) ; Anderson 2, fl. Oct. 1861 (OAL,- MEL) ; Kurz 2961, fl. (CAL); Maingay 2624A ((Kew Distr. 4), fl. & fr. 1867 (BM) ; Maingay 2624 (Kew Distr. 4), fl. 1867—8 (K) ; Kuntze' 6094, fl. Oct. 1875 (NY) ; millett 61, fl. 1884 (K) ; Ihwilund (CAL) ; Jurong, Ridley 435, fl. Oct. 1889 (CAL, SING) ; Eidley, fl. 1S90 (UC) ; Tangkei, Eidley, fl. & fr. Jan. 1905 (MO) ; Eidley, fl. & fr. (CAL, SING); Pulau Ubin, Ridley, fl. (SING).

Java: W. Java: Floem, fl. (L) ; Regantang, Boerlage, fr. June 1888 (L) ; Waning Manga, Kedunghalang, Boerlage 157, fr. Oct. 1888 (L) ; Buitenzorg, Boerlage, fl. & fr. Dee.. 1888 (L) ; ibidem, lldtticr Sa, fl. & fr. Apr. 1893 (BZ) ; Djasinga, Baclcer 9903, fl. Nov. 1913 (BZ) ; ibidem, Backer 10056, fl. & fr. Nov. 1913 (BZ) ; l-\ _ 1 T k !.. sn m -4 et ti ei T i , i / r*.r# \ n e r\ • • û i ,n et s\ /n .r* /• i n « n E of Dcpok, Backer 23168, fl. Jan. 1918. (®Z) ; S of Djasinga, Backer 26022, fr. 1918 (BZ) ; E of Depok, Baoker 26285, fl. Oct. 1918 (BZ) ; Dcpok, „ ,, _ ... _ _ r , , ... ... v ,, r , Bakhuizen van den Brink Jr 890, fl. & fr. Nov. 1921 (BZ, CAL, Gr, K, L, P, SING); Tegal Sapi, Bakhuizen van den Brink Jr 1568, Aug. 1922 (BZ) ; Djambu near Leuwiliang, Bakhuizen van den Brink 7830, fl. & fr. Aug. 19.11 (BZ, IC, L, U) ; Tendjoleat near Bunar, de Voogd 5, fl. Oct. 1940 (BZ) ; Land Bolang near Bunar, van Stccnis 12681, fl. Nov. 1941 (BZ) ; Parung Pandjang, Broekhuizen 15, fl. & fr. Dec. 1944 (RZ) ; ibidem, Broclchuizen 9, Feb. 1945 (BZ, L) ; ibidem, Broekhuizen, fr. Feb. 194i5 (BZ).

Borneo: (OAL, U) ; Korthals, fl. & fr. (CAL, K, L, MEL, S) ; Ü. S. S. Pac. Expl. Bxped. 18S8—13, fl. (NY) ; Burbidge, fl. (IC). — NU'. Borneo: Kuching, Bcccari PB 175 4- 193, fl. & fr. July 1865 (Fl, K, P) ; Brunei, Beeoari PB 4067, fl. Aug. 1867 (Fl) ; Rajang, Sibu, Ifavilatid = 2100, fl. Nov. 1892 (UC) ; near Kuching, lin Vila nd 2100, fl. & fr. Doc. 1892 (K) ; ibidem, JIaviUmd if- Hose = 2160, fl. & fr. Oct. 1894 (DM, DZ, CAL, CGE, GH, K, L, P) ; Kuehing, Eidley, fr. Aug. 189!) (SING) ; Sarawak, A nt. Coll. 557, f!. (A) ; Kapit, Upper Rajang Riv., Clemens 21167, fl. May 1929 (A, DZ, K, MO, NY). — IV. Borneo: Chaper, fr. (P) ; Sintang, Langlassê 68, fl. Juno 1894 (P) ; ibidem, Langlassé 87, fl. June 1894 (G, P) ; Acr Itam N of Pontianak, Folate 680, fl. Oct. 1940 (BZ, L) ; Pontianak, Enoh 200, fl. Sept. 1948 (BZ, K, L). — S. and SE. Borneo: Martapura, Korthals, fl. (L) ; Pulau Lampei, Korthals, fl. & fr. (L) ; Tewingan near Martapura, Labohm 1180, fl. Juno 1SHS (BZ) ;

Asem near Pleihari, Lahohm 1056, f 1. May 1!U!> (BZ) ; Bantau, Antasan Mastam, -*— — 7 "7 "*• V— y I J-WIU tau, XX41IUIVOIWI XliUOl/CWH, DG oh" lan 17, fl. Aug. 1922 (BZ). — E. and NE. Borneo: Sungai Rontanan, Bontang, Kutten 417, fr. March 1911 (BZ, U) ; Sungai Berbas, Bontang, Kutten 496, fl. & fr. May 1911 (U); Tikung, A m d jali 917, fl. & fr. Nov. 1912 (BZ, L, UC) ; Amdjali 1017, fl. 1912 (BiZ); near Sadjan Riv., Bulongan, Kutten 88, fr. 1914 (U). — Br. N. Borneo: Kudat, Fraser 120, fl. June 188ü (K) ; Sandakan, Creagh, fl. Apr. 1895 (K) ; Kinabalu Region, Gib bs 2930, fl. 1910 (BMj; Sandakan, Clemens 9504, fl. Doc. 1915 (A); TVood 458, fl. May—Aug. 1917 (K) ; Yates 56, fr. Oct. 1917 (A, US); Sandakan & Vicin., Wood 794, fl. & fr. Feb.—March 1920 (A, BZ, L, US); Sandakan, Mvburgh Pro v., Elmer 20104, fl. & fr. Oct.—Dec, 1921 (A, BM, BZ, C, OAB, X, L, M, MO, NY, P, S, SING, U, UC); Kg Mengalong, Weston, Suleiman BNB 2221, fr. Apr.' 1932 (BZ, IFI, K) ; Talaga, BalajaMa BNB 2561, fl. Aug. 1932 (A, BZ, IFT, K) ; Mt Kalawat, Kinabalu, Clemens 51317, fl. Jan. .1934 (BiM) ; Kabili For. Res., Sandakan, Puasa BNB 4846, fl. & fr. June 1935 (SING) ; Jambongaii Isl., Cabilmg BNB 3776, fl. (U-C) ; Tiaggau Ri»v., Keith BNB 9091, fl. June 1988 (SING) ; Marudu, Kudat, Kitaku Fior. Rtes., Austin BNB A1182, fl. Feb. 1948 (SINÎG) ; Elopura, Sandakan, Anthony BNB A 788, fr. March 1948 (SING) ; ibidem, Kadir BNB A[ 901, fl. Dec. 1948 (SING); ibidem, Kadir BNB A @691, fl. & fr. (L). — Baiiguey Isl. : „ 7 , __ t . Castro $■ Mele- (jrito 1502, fl. Sept. 1923 (A, UC, US). — Labuan: Low 192, fl. & fr. 1867 (Fl) ; Treacher, 1880 (S). — Anambas & Natuna Isls: near Genting, Sedanau Isl., van Stcenis 1055, fr. Apr. 1928 (BZ, L).

Philip piffles: hoher 2O , fl. (K). — Luzon: Mt Makiling, Laguna Prov., Quisumhmg, fr. Sept. 1925 (UC).

Cultivated: Bot. Gard. Bogor (origin : Banka) : Teysmawa, fl. & fr. 1800 (10; Kars 425, fl. (OAL) ; Kurz, fl. 1863 (P) ; Teysmamn, fr. 1867 (L) ; Beecari, fl. 1876 (Fl) ; Boerlage, fl. (L); no IV-G-18, fl. Nov. 1889 (BZ) ; J arise, 1899 (GRO) ; no IV-G-18, fr. 1903 (GH, NY, US) ; van Ilarreveld, fl. & fr. Sept. 1907 (GRO, L) ; Backer, fl. 1908 (BiZ) ; fl. & fr. Apr. 1912 (BZ) ; no IV-G-18a, fr. 1916 (BZ) ; Fevrell tf- Heide, fl. & fr. Dee. 1921 (S). —• ICultuurtuin Bogor: fl. & fr. (U) ; Giesmfiagen, fl. Feb. 1900 (M) ; no A-III-738, Idris, fl. & fr. May 1924 (WAG). — Victoria Garden, Bombay : Land, fl. Dee. 1935 (CAL). — Jamaica: Castleton Distr. (escape from Bot. Gard.), Harris 10889, fl. March 1910 (K, NY, U). — Public Gardens, Kuala Lumpur: Strugnell CF 126%%, fl. Jan. 1927 (SING). — Manila: Manila Gardens, Quimmbing BS 84737, fl. (NY); Fcnix 92, fl. & fr. Apr. 1938 (A) ; College of Agric., Laguna, Sulit PNI1 6.977, fl. & fr. Dee. 11)47 (A). — Panama Canal Zone Èxper. Gard.: JVetmore <)• Abbe 228, fl. Jan. 1932 (A). — Bot. Gard., Penang: Niir, fl. S;ept. 1918 (SING). — Bot. Gard"., Peradeniya: Faircliild Dorsett 262, fl. Feb. 19i26 (UC). — Royal Lakes Gard., Rangoon: Parkinson 14029, fl. March 1932 (DD) ; Parkinson 14455, fl. Juno 193:2 (DP). — School Gard., Semarang: Dooiers van Leeuwen-Eeynvami, fl. June 1910 (BZ). — Sierra Leone: Lanc-Poole 118bis, fl. June 1912 (K). — Bot. Gard., Singapore: fl. (A) ; llullett, fl. & fr. Aug. 1885 (SING) ; Curtis, ƒ]. Sept. 1900 (SING) ; Sargent, fl. Oct. 1003 (A) ; Lawn T, Nur, fl. Aug. 1918 (UC) ; Nur, fl. May 1919 (UC) ; Lakeside, N'ur, fl. & fr. Aug. 1920 (CAL, SING, UC) ; Arboretum, Nur, fl. June 1924 (SING) ; Kiah, fl. May 1929 (IFI) ; Lawn G, Nur, fl. Oct. 192,9 (BRI, BZ, SING) ; Chiliens M50&, fl. & fr. Nov. 1929 (NY, P) ; Furtado SF 34822, fl. Feb. & Sept. 193-8 (A, DO, IFI, L, SING). — Bot. Gard., Trinidad: Broadway, fl. May 1927 (IFI).

Distribution. Sumatra, Malay Peninsula, Riouw and Lingga Archipelagos, Banka, Billiton, Borneo, W. Java, Philippines. Prom Sumatra the species is known only from Palembang. The two collections from the Philippines may represent cultivated or naturalized specimens; I strongly suspect the species not to be indigenous in the Philippines. De Wit (1949) has pointed out that the species would be indigenous in Java. I do not agree with him for the following reasons: The first collecting of the species outside the Botanic Garden was done in 1888, i. e. about 30 years after its introduction. The first collections are all close near Bogor and later collections gradually come to farther distances. It is difficult to understand that a species like the present one with large and conspicuous flowers would have escaped the attention of earlier collectors, particularly as it flowers throughout the whole year in Bogor.

Ecology. In marshes, along streams, and on the margin of forests, sometimes immediately behind the mangrove, up to 500 m altitude Flowering continuously, each flower open for one day only; between the flowers of one raceme a difference in flowering-time of about 3—4 days. The fruit ripens after 36 days (Corner, 1940) ; the arillate seeds are eaten by birds, what may be a means of dispersal.

Vernacular names. Sumatra: Sempur ajor, S. rawah (Palembang) Simpong, Kaju sipur (Riouw and Lingga Arch.); Kembang masimpur (mesimpur), Minipor, Simpur, Simpor prampuan, Sipor (Banka) ;

Simpur (Billiton). Malay Peninsula: Champurna, Simpoh ayer (= water simpoh), S. gajah (i= elephant or big s.), S. pasir, S. paya (<= marsh s.). Borneo : Simpur (Mai., SE. Borneo), Simpor ayer (Mai., Br. N. Borneo), Simpor bini (Mai., Brunei), Simpor rimba (Kedayan, Br. N. Borneo), Tambakau (Tengara, Br. N. Borneo).

Uses. Because of its beautiful foliage and abundant flowering the species is often planted as an ornamental shrub.

Notes. 1. Wormia suffruticosa and W. subsessilis differ only in the indûment of the lower side of the leaves and petioles, this being villose in W. suffruticosa, glabrous in W. subsessilis. As several intermediates are known, the distinction of two separate taxa does not appearto be justified.

2. Wormia subsessilis var. borneensis differs from the other specimens by the large leaves and the different colour of the pseudocarp, this being white. As to the latter difference, I do not think it to be of any importance., whereas as to the other characters the pseudocarps in the type specimen of var. borneensis agree with those in other specimens. As to the size of the leaves, these are really large in the specimen cited, but intermediates as to size are found in a number of collections.

Asian Pacific Journal of Tropical Biomedicine

Ethnobotanical review and pharmacological properties of selected medicinal plants in Brunei Darussalam: Litsea elliptica, Dillenia suffruticosa, Dillenia excelsa, Aidia racemosa, Vitex pinnata and Senna alata

3. D. suffruticosa

Locally known as “Simpor bini”, D. suffruticosa is a medium sized tree characterized by its large bright flowers with five thin yellow petals around its white stamen, and dark-pink star-shaped fruits all of which are surrounded by large oval leaves (Figure 2). This plant species typically grows in wastelands, swamps, poor soil, white sands, secondary forests, along roadsides or the edge of forests [11–13]. Its leaves have been traditionally used for different treatments such as to promote wound healing, relieve rheumatism and treat fever while the fruit was claimed to be able to treat cancerous growths [12–16].

The methanolic extract of the roots of this plant have displayed significant antioxidant and cytotoxic activities particularly towards the HeLa cervical cancer cell line [17]. Studies conducted by Armania et al. have indicated that the phenolic content was an important contributor to the high antioxidant activity observed in the methanolic root extract of this plant species [17,18]. Although this extract showed the highest antioxidant and cytotoxic activities in the Hela cell line, it was found that the dichloromethane and ethyl acetate extracts exhibited higher cytotoxicity in the breast cancer cell lines, MCF7, MDA-MB-231, the A549 lung cancer cell line and the HT29 colon cancer cell line [17]. Further mechanistic investigation demonstrated that the plant extract inhibited the proliferation of the HeLa cervical cancer cell line as well as the MCF7 and MDA-MD-231 breast cancer cell lines via the

induction of apoptosis and the G2/M cell cycle arrest [17,18]. An in vivo study conducted by Yazan et al. showed that oral intake of aqueous root extracts have successfully reduced breast cancer induced in rats and also inhibited metastasis of the cancer to the heart [19]. The study further demonstrated that the extract was not toxic at the acute toxicity level up to a high dose of 500 mg/kg, however, mild focal hemorrhage was observed when a dose of 1000 mg/kg of the extract was used for treatment [19]. The cytotoxic activities of this plant species could be attributed to the presence of phytochemicals such as saponins, triterpenes, sterols, and polyphenolic compounds [17,20–26].

Saponins is a collective term for triterpenoid and steroidal glycosides [27], which consists of at least 150 kinds of natural saponins that have displayed significant anti-cancer properties [28]. Moreover, they have been recognized for their ability to reduce cholesterol level in the blood [29]. Dietary and endogenous cholesterols pass through the bile or desquamatedintestinal cells and reach the intestine before they are absorbed into the blood stream [29]. Saponins, being poorly absorbable from the intestine into the blood stream, interact with cholesterols and other sterols, and thus interfere and prevent them from being absorbed into the blood stream [29]. Additionally, saponins have the ability to stimulate the immune system and enhance antibody production [30].

Interestingly, multiple studies have reported reduction in bone loss with diets that are high in saponins [31,32]. One of the many saponins in particular, called asperosaponin VI, was able to induce the differentiation and maturation of osteoblasts and thus increase bone formation via the bone morphogenetic protein-2/p38 synthesis, and activation of the extracellular signal-regulated kinase 1/2 pathway [32].

On the other hand, polyphenols, which include phenolic acids, flavonoids, stilbenes and lignans, have been recognized for their multiple health-benefiting properties [33]. Polyphenols such as resveratrol and quercetin were found to exhibit significant cardio-protective effects by preventing platelet aggregation, disrupting atherosclerotic plaques and inhibiting protein expressions [33]. Other than that, anti-cancer, anti-viral, anti-diabetic, anti-aging and neuro-protective effects were also noted for the phytochemicals in the polyphenol group. They also showed beneficial effects towards asthma, osteoporosis, bone loss, skin damage and mineral absorption in intestines [33].

The phytochemical contents within D. suffruticosa play a significant role in relieving and alleviating illnesses. Perhaps it is a combination of their effects that promote wound healing, relieve rheumatism, and treat fever and cancerous growth traditionally. However, more studies should be performed to fully validate their traditional uses for such diseases.

ABSTRAK

Nama                   :    Suyatno

Program Studi     :    S1 Farmasi

Judul                    :    Uji Aktivitas Antibakteri Ekstrak Etanol 70% Daun Sempur (Dillenia suffruticosa (Griff.) Martelli)  Terhadap Bakteri Staphylococcus aureus Dan Escherichia coli.

Telah dilakukan penelitian tentang uji aktivitas antibakteri ekstrak etanol 70% daun sempur (Dillenia suffruticosa (Griff.) Martelli) terhadap bakteri Staphylococcus aureus dan Escherichia coli. Penelitian ini bertujuan untuk mengetahui aktivitas antibakteri dan nilai konsentrasi hambat minimun dari ekstrak etanol 70% daun sempur. Bahan uji yang digunakan adalah daun sempur yang diperoleh dari Desa Pedindang, kecamatan Pangkalanbaru, kabupaten Bangka Tengah, Propinsi Kepulauan Bangka Belitung untuk selanjutnya diproses menjadi serbuk halus. Pembuatan ekstrak dilakukan secara maserasi dengan etanol 70%, kemudian di evaporasi menjadi ekstrak kental dengan hasil rendemen sebesar 24,74%, hasil ekstrak dilakukan pengujian aktivitas antibakteri dengan konsentrasi 5%, 10%, dan 15% terhadap bakteri uji. Selanjutnya dilakukan pengukuran diameter daya hambat, berdasarakan pengukuran DDH  terhadap Staphylococcus aureus dengan hasil 5%  10,52 mm , 10% 11,73 mm, dan 15% 12,67 mm sedangkan terhadap Escherichia coli 5% 11,29 mm, 10% 12,89 mm, dan 15% 13,84 mm. Nilai KHM pada Staphylococcus aureus 3% dan pada  Escherichia coli 5% .

Kata Kunci : Ekstrak Etanol 70% Daun Sempur, Staphylococcus aureus, Escherichia coli, DDH, KHM.

Dilleniae radix (Dillenia suffruticosa, L. Dilleniaceae) the cytotoxic effect of dichloromethane extract at the maximum concentration of 100 μg/ml was applied to the breast cancer cell line (MCF-7). The experiment was conducted using the MTT assay in the presence of tamoxifen as the positive control. The extract showed the time and dose dependent strong cytotoxic activity with an IC50 value of 15.5 ± 0.5 μg/mL after 72 hours of exposure. This significant result indicates that the Dilleniae radix extract has a potential for developing a new cytotoxic agent. In addition, the authors explained how the investigated extract exhibited the cytotoxic effect. The dichloromethane extract of Dilleniae radix induced the G0/G1 and G2/M phase cell cycle arrest and apoptosis in caspase-3 deficient MCF-7 cells probably via the up-regulation of NF-κB, JNK1 and down regulation of the anti apoptotic genes AKT1 and ERK1 (70). Additionally, other studies confirmed the signalling pathway responsible for cytotoxicity of Dillenia suffruticosa extract on breast cancer cells (71, 72). Qualitative phytochemical screening of Dillenia suffruticosa extracts indicated the presence of saponins, triterpenes, sterols, and polyphenolic compounds, which contribute to the cytotoxic activities (72).

Journal of BIOLOGICAL RESEARCHES

ISSN: 08526834 | E-ISSN:2337-389X

Volume 22| No. 2| June | 2017

http://dx.doi.org/10.23869/bphjbr.22.2.20177

Published by © PBI East Java. Open Access  www.berkalahayati.org 74

 Corresponding Author:

Ridesti Rindyastuti

Purwodadi Botanic Garden, Indonesian Institute of Sciences.

Jl. Raya Surabaya-Malang Km. 65, Purwodadi, Pasuruan, Jawa Timur Phone: 0343-615033 Fax: 0341-426046

e-mail: ride17@gmail.com

Original Article

Carbon storage of medium-sized tree: a case study on Dillenia collection in Purwodadi Botanic Garden

Ridesti Rindyastuti

Purwodadi Botanic Garden, Indonesian Institute of Sciences Abstract

Abstract

Dillenia is a medium-sized tree which has high species diversity in tropical regions especially in Southeast Asia. Dillenia in Purwodadi Botanic Garden are collected from native habitats in Java, Kalimantan, Sulawesi and Papua which planted on the area of 17 x 55 m2. The purpose of this re-search is to study the above ground carbon storage in Dillenia collection in Purwodadi Botanic Garden. Carbon storage estimation was established by measuring stem carbon stocks from plant collections with plant age ranging between 12-30 years. Twelve years old collection contributed carbon storage of 7.35 tonnes/ha for D. sumatrana. Twenty years old species had the lowest carbon storage of 2.17 kg/plant for D. serrata and the highest of 51.9 kg/plant for D. auriculata with a range of carbon storage of 3.47 to 41.072 tonnes/Ha. Thirty years old plant contributed 39.465 kg/plant and carbon storages of 63.14 tonnes/ha for D. serrata and 135.59 kg/plant and 216.94 tonnes/ha for D. philipinensis. Overall, Dillenia collections in Pur-wodadi Botanic Garden contributed 793.94 kg carbon storages, store carbon on average of 30.54 kg/plant and 46.46 tonnes/ha. The increase of carbon storage in the second 10 years was higher than in the first 10 years. It indicated that Dillenia had growth strategy in the early growth then alocated more mass after 10 years. Carbon storage of Dillenia was high and different in age. D. serrata, D. papuana and D. auriculata are recommended species as a priority in planting trees based on carbon sink.

Keywords: Dillenia, carbon storage, medium-sized, tree, Purwodadi 

Received: 09 March 2017 Revised: 28 April 2017 Accepted: 08 June 2017

Introduction

According to Hairiah et al. (2011), carbon sequestra-tion is the ability of a system to store carbon from the atmosphere during a certain period. Plants through the process of photosynthesis have a function as an absorber of carbon emissions in the atmosphere and store it in the form of biomass. Generally, there are two kinds of carbon sequestration, which is below ground such as by soil microbes and roots, and above ground plant biomass mainly by tree stems (De Jong dkk., 1995; Cannell, 2003; IPCC, 2006). Climate change mitigation innitiatives recommend the reforestation programs and diversification of plant species for improving the ecosystems quality of degraded areas and for greening urban areas (IPCC, 2006; UN-FCCC, 2007). Selection of plant species in agroforestry programs based on carbon sink is important because plants could run various ecological functions and increase plants diversity as well (Diaz, 2009). Native species was proved of running more diverse environmental services compared to non-native species. In addition to its function as a carbon sink, native species are able to maintain the hydrology of an area, restore the food chain and native vegetation by associating with other local species.

Researches on carbon stocks of diverse plant species or groups, both native and non-native species in their nat-ural habitat have been carried out. Siregar and Darmawan (2011) examined the carbon sequestration of Dipterocar-paceae in Central Kalimantan, and argued that the dipter ocarp forests can store carbon 928.86 tonnes C/ha or 20.64 tonnes/plant. In the dipterocarp family, the varia-tions of their carbon storages are quite high. This can be caused by different species constituent, genetic factors that influence the allocation of nutrient storages and allocations, plant age and environmental factors (Diaz, 2009). Imiliyana et al. (2012) reported that mangrove is capable of storing 232.59 tonnes C/ha while according to Pramudji (2011) Acacia is capable of storing 56.05 tonnes C/ha.

Dillenia is one of medium-sized tree which has high species diversity in the tropics. This plant group has about 60 species distributed from Madagascar to Australia and is one of a vegetation component of tropical forest in low land areas. The leaves are oval to elliptical with promi-nent leaf midrib. The flowers are large with five petals and many stamens (Lemmens and Wong, 1995). Dillenia is utilized for many economic purposes such as wood products (most of Dillenia species), craft, and medicine such as Dillenia suffruticosa for anti-inflamatory (Shah et al., 2015), D. philiphinensis and D. indica for antimicrobial (Ragasa, 2009; Apu et al., 2010). Many previous studies on tree carbon storage did not use the growth size as a diagnostic characters to distinguish carbon stored in plants. Growth size of trees is controlled by gene and it influences common maximum tall and large of tree stem in which carbon stored. A group of trees which have a characteristic as medium-sized tree commonly grow to around 10-40 m tall, while group of large trees can grow up to 50 m tall even hundred m tall (Lemmens and Wong, 1995). The study on carbon storages of local plant species based on its growth size can increase the understanding of biology of trees and reveal the significant contributions of diverse plant species to carbon sequestration in reducing global carbon rising in the atmosphere.

Heng and Onichandran (2014), reported that Dille-nia suffruticosa in degraded area in Sarawak, Malaysia has slightly low biomass and carbon storage in the early forest formation (5.2 tonnes/ha). However, the highest proportion of biomass on this species was on the stem, and it increased with tree size and age. Dillenia can be a good recommendation for afforestation for both conserva-tion and tree planting in urban areas, but its carbon storage have not been studied well. Therefore, the aim of this research is to study 1) carbon storage of a medium-sized tree in a case of Dillenia collection in Purwodadi Botanic Garden and 2) the effect of different age on the carbon storage of Dillenia collection in a garden system, especially in Purwodadi Botanic Garden.

Method

Plant Materials

Dillenia collection examined in this study comprised of 10 species. The collections were planted on an area approximately of 17 x 55 m2. List of plant species studied together with the origin of the collection and distribution of the species are presented in Table 1.

Table 1. Species list, origin of plant collection and plant distribution of Dillenia collection in Purwodadi Botanic Garden.

(Source : Catalog of plant collection in Purwodadi Botanic Garden-LIPI, 2012; Lemmens and Wong, 1995)

Biomass measurement

Biomass was measured using allometrics method which can be estimated from stem diameter at breast height of adult trees so called DBH and trees height of each Dillenia collection in Purwodadi Botanic Garden-LIPI. Three replications were used in the measurement for each species. Biomass was obtained by calculating using the formula of biomass for plants in dry climates habitat and allometric equation for Dillenia as follows:

(Hairiah et al., 2010)

Biomass (kg) = 0,122 (rD2H)0.916

Where r = wood density (Zanne et al., 2009)

D = diameter (cm)

H = plant height (m)

Carbon stock values were obtained by multiplying the values of biomass with allometric values for carbon stock i.e. 0.46 (Hairiah et al., 2010). Carbon storage per hectare was converted using the total of area and plant spacing of collection in Purwodadi Botanic Garden i.e. 2.5 x 2.5 m.

Data Analyses

Biomass and carbon stocks of 10 Dillenia species were analyzed using the variance test (ANOVA) at the 95% confidence level to determine the variation of bio-mass and carbon stocks among species in Dillenia collec-tions. The data analyses were also conducted on the bio-mass and carbon stocks among species in different age level (12, 20 and 30 years).

Results

Carbon Storage in Dillenia

Table 2 showed the carbon storage of Dillenia collection in Purwodadi Botanic Garden estimated from their DBH and biomass. The statistical analyses are also conducted to show the differences of carbon stocks among Dillenia species. Based on the ANOVA test on a 95% confidence level, DBH of 10 species of Dillenia were significantly different amongs species with a P value of 0.004 (P <0.05). The statistical tests were conducted on the DBH of individuals in various age ranges (12-30 years). The data of biomass and carbon stocks were ab-normal based on the normality test on a confidence level of 95%. Therefore, the data only can be read descriptive-ly. The 30 years old D. philipinensis have the highest DBH while the twelve years old D. sumatrana has the lowest DBH. The species which are 20 years old have a range of DBH between 18.15 cm for D. ovalivolia to 54 cm for D. papuana. It showed that in the same age, plant species within genus may have different DBH and carbon storage.

Table 2. The value of DBH, biomass, per plant and per ha carbon stocks of 10 species of Dillenia in Purwodadi Botanic Garden.

*) Statictical analyses using ANOVA in the confidence level of 95%. Same letters in one column showed no significant  ifference among species 

**) These data are not normal and not random based on the normality and randomity test, thus the variance analysis can not be performed.

D. philipinensis has the highest biomass (29.47 kg) and contribute the total carbon storage of 135.58 kg C/plants or 216.94 tonnes C/ha. Twelve years old D. su-matrana has DBH 21.93 cm with 0.99 kg of biomass and carbon stocks of 4.59 kg/plant or 7.35 ton C/ha. This size is quite large compared to other 20 years old species (Fig 1 and Table 2). Figure 3 showed the carbon stored per hectare of Dillenia collection in Purwodadi Botanic Garden. Carbon storage of D. ovalifolia is the lowest among other species studied especially compared to other species at the same age (20 years old). At 20 years old, D. papuana has the highest carbon storage of 51.9 kg C/plant.

Medicinal uses, chemistry and pharmacology of Dillenia species (Dilleniaceae)

1. Introduction

The genus Dillenia belongs to the Dilleniaceae family and contains approximately 100 known species (Lim, 2012). According to The Plant List (2013), as many as 175 scientific plant names from the genus Dillenia have been recorded with 58 accepted names (low and medium confidence levels) and 71 names of synonym species.
The name Dillenia is derived from Joannes Jacobus Dillenius, a British botanist who dedicated his efforts in the field of taxonomy of this genus (Quattrocchi, 2012). Dillenia species are monoecious plants which produce attractive flowers and yellow fruits. D. indica is known for its lemon-flavored fruits that are use to make jellies and curries. These species are evergreen and deciduous trees or shrubs of disjunct distribution in the seasonal tropics of Madagascar through South and South East Asia, North Australia, and Fiji (Dickison, 1979; Horn, 2007; Kerrigan et al., 2011; Lim, 2012). They grow from sea level to an elevation of about 2000 m. The plants also grow in forests, and several species show an adaptation to temporary flooded situations. They are mostly trees that form large leaves and flowers in few-flowered inflorescences (Dickison, 1979). Their barks are unique in fine colors of red, grayish, and reddish brown that are used in furniture making (Hoogland, 1952). Several species of this genus produce sweetishsour and astringent edible fruits (Hoogland, 1952; Jansen et al.,
1992; Kerrigan et al., 2011; Lim, 2012; Saha and Sundriyal, 2012) and are cultivated as ornamental plants (Hoogland, 1952; Kerrigan et al., 2011).
Based on our extensive search regarding medicinal uses, chemical constituents, and biological activities of the species from the genus Dillenia, only very few species have been described thus far. Starting from 1962 to the present, 19 species of the genus Dillenia have been reported for their medicinal uses and their phytochemistry. These 19 species are Dillenia andamanica C.E.Parkinson, D. aurea Sm., D. bracteata Wight, D. excelsa (Jack) Martelli ex Gilg., D. indica L., D. ovata Wall. ex Hook.f. & Thomson, D. papuana Martelli, D. parviflora Griff., D. pentagyna Roxb., D. philippinensis
Rolfe, D. pulchella (Jack) Gilg., D. reticulata King, D. retusa Thunb., D. scabrella (D.Don) Roxb. exWall., D. eximia Miq., D. serrata Thunb., D. suffruticosa (Griff.) Martelli, D. sumatrana Miq., and D. triquetra (Rottb.) Gilg. However, out of these, only 7 species have been evaluated for their biological activities. As part of our search for natural anti-inflammatory compounds, three triterpenes have been isolated from D. serrata and they performed pronounced inhibitory activity on the production of prostaglandin E2, which is known as a predominant inflammatory mediator (Jalil et al., 2015). This review describes the current state of their medicinal properties, chemistry, and pharmacological aspects.

4.1. Antimicrobial activity

Some of Dillenia species were investigated for antibacterial, antifungal, and antiviral activities. The extracts and fractions of D. indica, D. papuana, D. pentagyna, D. suffruticosa, and D. sumatrana were reported to possess growth inhibition against Gram positive and negative bacteria (Table 5). However, they were found to exhibit weak growth inhibition against tested fungi, including Aspergillus fumigatus, A. niger, Candida albicans, C. arriza, C. crusei, Penicillium sp., Rhizopus oryzae, Saccharomyces cerevisiae, and Trichoderma viride (Nick et al., 1995a; Wiart et al., 2004; Haque et al., 2008; Apu et al., 2010; Smitha et al., 2012). Seven triterpenoids (37, 42e45 and 47e48) from Dillenia plants were proven to have antimicrobial action (Nick et al., 1994, 1995b; Ragasa et al., 2009).

Strong growth inhibition against Escherichia coli and Bacillus subtilis has been shown by 45 that was assayed using the bioautographic method on TLC plate. Meanwhile, 48 was the most active against the growth of Micrococcus luteus (Nick et al., 1995b). Similar pronounce growth inhibition of 48 also reported towards E. coli, Pseudomonas aeruginosa, Staphylococcus aureus, and B. subtilis (Ragasa et al., 2009). These findings suggested that Dillenia plants traditional uses for the therapeutic remedies of microbial infectionrelated diseases such as diarrhea, dysentery, septicaemical infection and skin-related diseases had a potential as antimicrobial agent, which supported their traditional uses for the therapeutic remedies of microbial infectionrelated diseases such as diarrhea, dysentery, septicaemical infection and skin-related diseases

Antibacterial and antifungal

The leaves extract inhibited the growth of B. cereus, B. subtilis, P. aeruginosa, and C. albicans with zones ranging from 7 to 9 at 1 mg/disc as compared to gentamycin (10 mg/disc; 18e20 mm for B. cereus, B. subtilis, P. aeruginosa) and nystatin (20 mg/disc; 11 mm for C. albicans).

The effect of herbal plant extracts on the growth and sporulation of Colletotrichum gloeosporioides

Johnny et al. .J. Appl. Biosci. 2010.                                                                                    The effect of herbal plant extracts on Colletotrichum gloeosporioides

Journal of Applied Biosciences 34: 2218 - 2224

ISSN 1997–5902

The effect of herbal plant extracts on the growth and

sporulation of Colletotrichum gloeosporioides

Lucy Johnny*, Umi Kalsom Yusuf, and Rosimah Nulit

Department of Biology, Faculty of Science, University Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan,

Malaysia.

*Author for correspondence: lucyjohnny13784@yahoo.com, Tel. +6017-3434717; Fax: +603-86567454

Original submitted on 22th July 2010. Published online at www.biosciences.elewa.org on October 7, 2010

ABSTRACT

Objectives: The antifungal activities of the leaf extracts of fifteen selected medicinal plants; Alpinia galanga L., Alstonia spatulata Blume., Annona muricata L., Blechnum orientale L., Blumea balsamifera L., Centella asiatica L., Dicranopteris linearis, Dillenia suffruticosa, Litsea garciae Vidal., Melastoma malabathricum L., Momordica charantia L., Nephrolepis biserrata (Sw.)., Pangium edule Reinw., Piper betle L., and Polygonum minus Huds., were evaluated on the plant pathogenic fungus; C. gloeosporioides isolated from mango.

Methodology and results: Different antifungal assays were employed, i.e. Agar-Disc Dilution assay as

primary screening assay, followed by determination of Minimum Inhibition Concentration (MIC), and the rate of sporulation assay. The antifungal assay was carried out in Potato Dextrose Media in five different treatments, i.e.; distilled water as negative control, crude extract of leaves in methanol, chloroform, acetone and Benomyl as positive control. A. galanga extracts were most effective and exhibited highest antifungal activities against C. gloeosporioides. Methanol crude extract reduced radial growth of C. gloeosporioides by 66.39%, followed by chloroform crude extract 63.26%, and 61.56% for acetone crude extracts. The exact concentrations that have definite potential to fully restrict the growth of C. gloeosporioides (MICs) for A. galanga is 15.00 mg/mL in methanol, 17.50 mg/mL in chloroform, and 17.50 mg/mL in acetone. The sporulation assay also revealed that A. galanga leaves crude extracts showed highest inhibition of spore germination of C. gloeosporioides overall at concentration of 10 mg/mL; with 68.89% inhibition by methanol extracts, 64.13% by chloroform extracts, and 62.86% by acetone extracts.

Conclusion and application of findings: Numerous natural products of plant origin are pesticidal and have the potential to control fungal diseases of crops. Thus, considerable effort should be devoted to screening plants in order to develop new natural fungicides as alternative to existing. In this study, the leaf crude extracts of A. galanga exhibited effectiveness against C. gloeosporioides and should be considered for further evaluation.

Key words: Medicinal plants, crude extracts, anti-fungal growth, C. gloeosporioides

INTRODUCTION

Colletotrichum gloeosporioides are one of the most important pathogens affecting the flowers and fruits of mango trees causing anthracnose worldwide. In areas where rain is prevalent during flowering and fruit set, anthracnose can cause destruction of the inflorescences and infection and drop of young fruit where this can obviously lead to serious losses, reaching up to 35% of the harvested fruit (Martinez et al., 2009). Excessive use of benomyl, thiophanate-methyl and thiobendazole as pre- and post-harvest sprays has led to a reduction in effectiveness in certain areas where pathogen resistance to fungicides has been reported (Spalding, 1982). Indiscriminate use of the chemicals is not only hazardous to people but also disrupt the natural ecological balance by killing the beneficial soil microbes (Ansari, 1995). So, alternatives have to be developed to control anthracnose in order to guarantee safe food production as well as reduce environmental pollution. The integration of a number of practices aiming to reduce or eliminate negative side effects caused by chemicals used for controlling major mango diseases is the most realistic option for solving the problem (Chowdury and Rahim, 2009). Research work in relation to anthracnose disease management of mango is yet to develop effective alternative/options.

Hence, this study was carried out with the aim of providing broader options by evaluating the antifungal activity of crude leaf extracts from selected medicinal plants against phytopathogenic fungi Colletotrichum capsici and Colletotrichum gloeosporioides.

MATERIALS AND METHODS

The leaves samples were collected from Sarikei, Sarawak. Samples were dried in the oven at 50ºC until the leaves become crunchy and were weighed. 100 g of leaves samples were then pounded using mortar and pestle into coarse powder. Leaves of the plants were extracted in polar solvent (methanol), semi-polar solvent (chloroform), and non-polar solvent (acetone) by soaking in each solvent for at least 48 hours. The extracts solution was filtered with filter paper (Whatman No. 1), transferred into pre-weighed 250mL round bottom flasks and evaporated using Rotavapor. The extracts and the round bottom flask were weighed again after solvent evaporation.

Pathogen culture: The culture of Colletotrichum gloeosporioides from Mangifera india L. were obtained from Faculty of Agriculture, UPM. Pure cultures were maintained on Potato Dextrose Agar slants.

Antifungal assay: The antifungal assay was carried out according to Alam (2004) with a slight modification.

A volume of 19 mL of molten PDA was poured in sterilized Petri dishes along with 1 mL of plant extract and plated. Mycelial discs (10 mm diameter) made using the cork borer were inoculated at the centre of the medium. The antifungal assay was carried out in PDA in five different sets: negative control, crude extract of leaves in methanol, chloroform, and acetone, and positive control (commercial fungicide). Colony growth was measured on the basis of linear dimensions. Minimum inhibitory concentration (MIC) and sporulation were determined.

Sporulation was determined by adding 10ml sterile distilled water to each seven days old plate that were obtained from agar disk method and gently scraping the mycelia with a sterile glass rod to dislodge the spores.

The spore suspensions obtained were filtered through sterile cheesecloth into a sterile 50 mL glass beaker and homogenized by manual shaking. The spores were then counted using a haemocytometer.

The percentage reduction (Sr) or stimulation (Ss) of sporulation by each extract was determined using the following formula, (Nduagu et al., 2008):

Sr = (S1 – S2) x 100

              S1

Where;

Sr = Percentage of reduction in sporulation;

S1 = Sporulation on the untreated medium (control);

and

S2 = Sporulation on the treated medium.

Ss = (S2 – S1) x 100

              S2

Where;

Ss = Percentage of stimulation in sporulation;

S2 = Sporulation on treated medium; and

S1 = Sporulation in untreated medium.

RESULTS

Inhibition of radial growth of Colletotrichum gloeosporioides: Among the plants screened, only 5 species showed 50% or more antifungal activity against C. gloeosporioides at varying concentrations (Table 1).

Methanol crude extracts of A. galanga, P. betle, M. malabathricum, B. balsamifera, and P. minus were

most effective. A. galanga exhibited highest antifungal activity of 66.78% at 10.00 Hg/mL, 66.27% at 1.00 Hg/mL, 61.57% at 0.10 Hg/mL, and 60.55% at 0.01 Hg/mL against C. gloeosporioides. This was followed by M. malabathricum and P. Betle, whose inhibition activity increased as the concentration of plant extracts increased (Table 1).

Table 1: Inhibiton of radial growth (mm) of Colletotrichum gloeosporioides by varying concentrations of leaf extracts in methanol.

Extracts of A. galanga in chloroform also showed high inhibition against C. gloeosporioides of 63.69% at 10.00 Hg/mL, 61.43% at 1.00 Hg/mL, 60.78% at 0.10 Hg/mL, and 58.62% at 0.01 Hg/mL. This was followed by chloroform crude extract of P. betle, M. malabathrichum, and B. balsamifera (Table 2). Extracts of A. galanga in acetone inhibited growth of C. gloeosporioides 62.60% at 10.00 Hg/mL, 60.50% at 1.00 Hg/mL, 57.05% at 0.10 Hg/mL, and 54.67% at 0.01 Hg/mL (Table 3).

Minimum Inhibition Concentration (MIC) of plant crude extracts to C. Gloeosporioides: Crude extracts of Alphinia galanga in all three solvents; methanol, chloroform and acetone exhibited the lowest MIC value against C. gloeosporioides of 15.0 Hg/mL in methanol, 17.50 Hg/mL in chloroform and acetone. This was followed by crude extracts of both P. betle and M. malabathricum which exhibited no visible growth of C. gloeosporioides at 17.50 Hg/mL and 20.00 Hg/mL. Crude extracts of C. asiatica, D. suffruticosa, A. muricata, D. linearis, A. spatulata, L. garciae, P. edule, and N. bisserrata in all three solvents exhibited MIC values that were out of the range of the prepared concentrations. Crude extracts of B. orientale did not exhibit any antifungal activities against C. gloeosporioides.

Table 2: Inhibiton of radial growth of C. gloeosporioides by varying concentrations of leaf extracts in chloroform.

Sporulation : The highest inhibition was recorded in treatments of crude extracts of A. galanga which exhibited the highest antifungal activities in inhibiting the sporulation of C. gloeosporioides among the 15 medicinal plants (Table 4-6). The methanol crude extract of A. galanga at 10.00 Hg/mL exhibited the highest inhibition overall of 68.89%. At the lowest concentration of 0.01 Hg/mL of acetone crude extract, A. galanga still inhibited sporulation by 62.86%.

Table 3: Inhibiton of sporulation (x105) of C. gloeosporioides by varying concentration of leaf extracts in acetone.

Table 4: Inhibiton of sporulation (x105) of C. gloeosporioides by varying concentration of leaf extracts in methanol.

Table 5: Inhibiton of sporulation (x105) of C. gloeosporioides by varying concentrations of leaf extracts in chloroform.

Table 6: Inhibiton of sporulation (x105) of C. gloeosporioides by varying concentrations of leaf extracts in acetone.

Each value represented the mean ± standard error; NI = No Inhibition, * represented crude extracts effectively inhibits growth

DISCUSSION

The results obtained from the different studies showed that the crude extracts of Alpinia galanga leaves exhibited the highest antifungal activities against C. gloeosporioides. The inhibition was expressed as reduced radial growth, dry mycelial weight, and sporulation germination rate of the pathogen. Inhibitory action of A. galanga crude extracts was recorded even at very low dose, which is a clear indication that the crude extracts contain active commencements that have antifungal properties.

Antimicrobial activity of extract from A. galanga could be attributed to its chemical constituents as reported by Saritnum and Sruamsiri (2003). Phongpaichit and Liamthong (2001) mentioned that chavicol acetate that has been identified as an antifungal component from A. galanga plays an antifungal role as in the Alpinia conchigera oil which has antifungal activity against C. gloeosporioides. The ethanol extracts from A. galanga have been found to have pronounced inhibitory activities against a wide variety of human pathogenic fungi, including strains resistant to the common antifungals amphotericin B and ketoconazole (Elfahmi, 2006). 1'-Acetoxychavicol acetate, the active compound from A. galanga has antifungal activity against Trichophyton mentagrophytes, T. rubrum, T. concentricum, Rhizopus stolonifer and Aspergillus niger at a concentration 14 mg/mL (Janssen and Scheffer, 1985). In agreement with this, Abad et al. (2007) mentioned that the chloroform extract of A. galanga has pronounced antifungal activity against Cryptococcus neoformans, Microsporum gypseum and Trichophyton longifusus. Many of the medicinal plants selected for this study have been reported to have novel bioactivities. Piper betle exhibited the best antifungal activities against C. gloeosporioides after Alpinia galanga. Johann et al. (2007) stated that the chemistry of Piper species has been widely investigated and phytochemical investigations from different parts of the world have led to the isolation of a number of physiologically active compounds such as alkaloids/amides, propenylphenols, lignans, neolignans, terpenes, steroids, kawapyrones, piperolides, chalcones, di-hydrochalcones, flavones and flavanones which exhibited high antimicrobial and antifungal properties. According to Lee et al. (2004), most Piper chemistry has been conducted to find potential pharmaceuticals or pesticides, and over 90% of the literature focuses on compounds that are cytotoxic, antifungal, antitumor, fragrant, or otherwise useful to humans.The leaves of Blumea balsamifera contain icthyothereol acetate and cryptomeridiol, lutein, and K-carotene (Ragasa, 2004). Antimicrobial tests indicated that icthyothereol acetate has moderate activity against Aspergillus niger, Trichophyton mentagrophytes and Candida albicans (Ragasa, 2004).

In conclusion, the crude extracts of leaves that exhibited good potential as fungicides of C. gloeosporioides should be evaluated further in in-depth to study the phytoextracts for their potentiality under in vivo condition.

ACKNOWLDEGEMENTS: Author acknowledges with gratitude the Graduate Research Fellowship (GRF) of Universiti Putra Malaysia.

REFERENCES

Abad MJ, Ansuategui M, Bermejo P, 2007. Active antifungal substances from natural sources. ARKIVOC. Vol. VII: 116-145.

Alam S, 2004. Synthesis, antibacterial and antifungal activity of some derivatives 2-phenyl-chromen-4-one. J. Chem. Sci. Vol. 116: 325-331.

Ansari MM, 1995. Control of Sheath blight of rice by plant extracts. Indian Phytopath. 48(3): 268-270.

Chowdury MNA. and Rahim MA, 2009. Integrated crop management to control anthracnose (Colletotrichum gloeosporioides) of mango. Journal of Agriculture and Rural Development 7(1&2): 115-120.

Elfahmi, 2006. Phytochemical and biosynthetic studies of lignans with a focus on Indonesian medicinal plants. PhD thesis. University of Groningen.

Janssen AM. and Scheffer JJC, 1985. Acetoxychavicol acetate, an antifungal component of Alpinia galanga. Planta Medica 6: 507-511.

Johann S, Pizzolatti MG, Donnici CL, Resende MA, 2007. Antifungal properties of plants used in Brazilian traditional medicine against clinically relevant fungal pathogens. Brazilian Journal of Microbiology 38: 632-637.

Khewkhom N, Shangchote S, Greger H, 2007. In vitro antifungal activity of some well-known spices against plant pathogenic fungi. Agricultural Sci. J. 38 (6) (Suppl.): 70-74.

Lee SW, Musa N, Chuah TS, Wee W, Shazili NAM, 2008. Antimicrobial properties of tropical plants against 12 pathogenic bacteria isolated from aquatic organisms. African Journal of Biotechnology. Vol. 7 (13): 2275-2278.

Martinez EP, Hio JC, Osorio JA, Torres MF, 2009. Identification of Colletotrichum species causing anthracnose on Tahiti lime, tree tomato and mango. Agronomia Colombiana. Vol. 27 (2): 211-218.

Nduagu C, Ekefan EJ, Nwankiti AO, 2008. Effect of some crude plant extracts on growth of Colletotrichum capsici (Synd.) Butler & Bisby, causal agent of pepper anthracnose. Journal of Applied Biosciences 6 (2): 184-190.

Nguyen NN, 2002. Screening of medicinal plants and essential oils having antifungal activity. Special Issue of Medical Research 6 (1) (Suppl.): 165-169.

Okwu DE, Awurum AN, Okoronkwo JI, 2007. Phytochemical composition and in vitro antifungal screening of extracts from citrus plants against Fusarium Oxysporum of okra plant (Hibiscus eculentus). African Crop Science Conference Proceedings. 8: 1755-1758.

Phongpaichit S. and Liamthong S, 2001. Antifungal activities of plant extracts against Colletotrichum gloeosporioides (Penz.) Sacc. J. Natl. Res. Council Thailand. 33 (1): 55-68.

Ragasa CY, 2004. Isolation, structure elucidation, and antimicrobial assay of secondary metabolites from five Philippine medicinal plants. The URCO Digest. VI (2): 3.

Saritnum O. and Sruamsiri P, 2003. Random amplified polymorphic DNA analysis of galanga (Alpinia spp.) accessions. CMU Journal 2 (3): 159-164.

Spalding DH, 1982. Resistance of mango pathogens to fungicides used to control post harvest diseases. Plant Disease 66:1185-1186.

Srinon W, Wuntid T, Soytong K, 2009. Antifungal activities from different solvent extracts of medicinal plants against Colletotrichum gloeosporioides (Penz.) causal organism of mango anthracnose disease. Agricultural Sci. J. 40 (1) (Suppl.): 75-78.

Suprapta DN. and Khalimi K, 2009. Efficacy of plant extract formulations to suppress stem rot disease on vanilla seedlings. J. ISSAAS. 15 (2): 34-41.

Yazdani D, Rezazadeh SH, Amin GH, Zainal AMA, Shahnazi S, Jamalifar H, 2009. Antifungal activity of dried extracts of anise (Pimpinella anisum L.) and star anise (Illicium verum Hook. f.) against dermatophyte and saprophyte fungi. Journal of Medicinal Plants 8 (5) (Suppl.): 24-29.