Antibacterial Screening of the Culture of Endophytic Fungal Extracts Isolated from Cinnamon Stick (Cinnamomum burmannii [Nees & T. Nees] Blume)
Praptiwi, Muhammad Ilyas, Ahmad Fathoni, Dewi Wulansari dan Andria Agusta*
Phytochemistry Laboratory, Research Center for Biology- Indonesian Institute of Sciences
Jl. Raya Jakarta-Bogor Km. 46, Cibinong 16911. *E-mail: andr002@lipi.go.id
ABSTRACT
Pathogenic bacterial resistance to antibacterial agents used in the treatment of recent formal systems have undergone significant increase. It is necessary to get a source of new antibacterial agents to overcome resistance. Endophytic fungi have been known as one of the organisms that have the ability to produce a variety of bioactive metabolites. In this study 26 endophytic fungi have been isolated from various parts of Cinnamomum burmannii plant. The results of antibacterial activity screening of endophytic fungal extracts by TLC-bioautography method without elution showed that as many as 18 extracts inhibit the growth of Staphylococcus aureus InaCC-B4 with the diameter of inhibition zone (DIZ) were in the range of 5.00 to 16.00 mm. On the other hand, the number of extracts that could inhibit the growth of Escherichia coli InaCC-B5 were 24 extracts with the DIZ were in the range of 4.00 to 28.00 mm. A detailed analysis of eluted TLC-bioautography method showed the chemical compounds of extracts that responsible for antibacterial activity.
Keywords : Cinnamomum burmannii, endophytic fungi, antibacterial, TLC-bioautography
ABSTRAK
Resistensi bakteri patogen terhadap bahan antibakteri yang biasa digunakan dalam sistem pengobatan formal telah mengalami peningkatan yang signifikan. Oleh sebab itu sangat penting untuk mendapatkan sumber bahan antibakteri baru yang dapat mengatasi resistensi. Jamur endofit telah dikenal sebagai salah satu organisme yang memiliki kemampuan untuk menghasilkan berbagai metabolit bioaktif. Dalam penelitian ini 26 jamur endofit telah berhasil diisolasi dari berbagai bagian tanaman Cinnamomum burmannii. Hasil skrining aktivitas antibakteri ekstrak jamur endofit dengan metode TLC-bioautografi tanpa elusi menunjukkan bahwa sebanyak 18 ekstrak menghambat pertumbuhan Staphylococcus aureus InaCC-B4 dengan diameter zona hambat (DIZ) pada kisaran 5,00-16,00 mm. Jumlah ekstrak yang menghambat pertumbuhan Escherichia coli InaCC-B5 adalah 24 ekstrak dengan kisaran diameter zona hambat antara 4,00-28,00 mm. Hasil elusi pada TLC-bioautografi terhadap ekstrak jamur endofit C. burmannii menunjukkan adanya beberapa komponen kimia yang mempunyai aktivitas antibakteri.
Kata kunci : Cinnamomum burmannii, jamur endofit, antibakteri, KLT-bioautografi
INTRODUCTION
Pathogenic microbial resistance to antimicrobial drugs lately was increased significantly, so it is becoming a global concern to overcome. Therefore, the search for new compounds as a source of antimicrobial drugs is very urgent. On the other hand, the presence of endophytic microbes are very abundant and capable of producing bioactive secondary metabolites[1]which can be used as a source of new bioactive metabolites as antimicrobial drugs.[2]
Endophytic fungi is defined as a fungus that grows in a healthy plant tissues without causing any signs of disease or damage to the host plant.[3] Endophytic fungi have the ability to produce the same active compound with a compound produced by the host plant.[4] According to Newman and Cragg[5] endophytic fungus has been considered as a useful resource in the search for new biologically active compounds, so that the endophytic fungus referred to as 'synthesizer' chemical compounds in plants.[6] Some extracts of endophytic fungi has been known to have the ability as an antimicrobial, insecticidal, antiviral, antitumor, and antioxidants.[7,8] The isolation of endophytic fungi to produce bioactive compounds is an efficient method in the search for new active compounds.[9] In the last two decades, many bioactive compounds from endophytic fungi have been found and are potentially as antimicrobial, insecticidal, cytotoxic, and anticancer.[4]
Cinnamomum is a genus of the Lauraceae family that are widely used as a spice and traditional medicine. Some endophytic fungi associated with Cinnamomum has been reported to have the ability to produce bioactive compounds. Muscodor albus is endophytic fungi isolated from Cinnamomum zeylanicum reported to produce volatile compounds that inhibit the growth of Escherichia coli and some fungi and yeasts.[10] Unidentified endophytic fungi associated with C. mollisimum produces 5-hidroxyamulocyn that inhibit the growth of Aspergillus niger and P388 cells (leukemia) with IC50 1.56 ug / ml and 2.10 ug / ml respectively.[11]
Isolation of endophytic fungi associated with various parts of cinnamon plant as well as the screening of antibacterial properties of these extracts will be reported in this paper.
MATERIAL AND METHODS
Plant material
Young stem and leaves of cinnamon were collected from Arau valley, Limapuluh Kota District, West Sumatra. Identification of plant was done in Herbarium Bogoriense –Botany Division, Research Center for Biology- Indonesian Institute of Sciences, Cibinong.
Bacterial isolates
Bacterial isolates (S. aureus InaCC-B4 and E. coli InaCC- B5) used for antibacterial test are collection of Indonesian Culture Collection (InaCC), Microbiology Division, Research Center for Biology- Indonesian Institute of Sciences, Cibinong.
Isolation and culture of endophytic Fungi
Samples of leaves and cinnamon stem from the field is stored at low temperature (cooler box) during transport, after arriving at the laboratory the samples were washed with tap water, then soaked in 70% alcohol for 2 min, followed by sterilization of 5.3% sodium hypochlorite for 5 minutes and then rinsed with 70% ethanol 0.5 minutes. The samples were surface sterilized and then cut into small pieces (1 x 1 cm2), the stems were splitted with a sterile knife. Each sample was dried under aseptic conditions. The sample is then placed on top of the Corn Meal Agar (CMMA) growth medium added with chloramphenicol 0.05 mg / ml, and then incubated at room temperature for 1 week. Endophytic fungal colonies were transferred on Potato Dextrose Agar (PDA) several times to get a single isolate.[12]
Identification of endophytic fungal isolates
Identification of fungal morphological characters based on the guidelines[13,14,15,16,17]. Identification is done by observing morphological traits and characteristics both macroscopically and microscopically from fungal colonies were grown on PDA at room temperature. Macroscopic characters observed were color and surface colonies (granular, such as flour, mounting, slippery), texture, zonation, growth area, the lines of radial and concentric, reverse color, and exudate drops.
Cultivation and extraction of endophytic fungal cultures
Endophytic fungal isolates were then cultivated in two types of liquid medium : Potato Dextrose Broth (PDB, 200 ml) and Glucose Yeast Peptone (GYP, 200 ml). After 3 weeks incubation under static conditions at room temperature, the media and their fungal biomass extracted three times with ethyl acetate and then concentrated with rotary evaporator.
Analysis of the chemical compounds of endophytic fungal extracts
The chemical compounds of endophytic fungal extracts were analyzed by Thin Layer Chromatography ( TLC , silica gel GF254 , Merck ) and eluted with a mobile phase dichloromethane : methanol ( 10 : 1 ) . Chromatogram was observed under UV light at wavelengths of 254 and 366 nm then sprayed with a stain reagent : 1 % Ce ( SO4 ) 2/10 % H2SO4 and 1 % vanillin / H2SO4.
Antibacterial assay
Antibacterial screening of ethyl acetate extracts of endophytic fungi associated with cinnamon plant was conducted by TLC-bioautography. 100 ug extract (10 mg / ml in acetone) spotted on TLC plates (Silica gel GF254, Merck), and then dipped into the bacterial suspension (E. coli InaCC-B5 and S. aureus InaCC-B4) with density of 106 cfu / ml in BHI (Brain Heart Infussion) liquid medium, Difco. Furthermore, the TLC plate was placed in a sterile petri dish and incubated at 37 ° C. After 18 hours of incubation TLC plates were sprayed with iodonitroptetrazolium p-violet (INT, Sigma). Observations were made 1 hour after spraying. Antibacterial activity is characterized by the formation of a white area on purple background.
Chromatogram of endophytic fungal extracts was used to determine the compounds responsible for the antibacterial activity. 100 ug extract (1 mg / ml in acetone) spotted on TLC plates (Silica gel GF254, Merck) and then eluted with mobile phase of dichloromethane - methanol (10: 1). The TLC plate was then dipped into the bacterial suspension and incubated for 18 hours. After incubation completed, the TLC plate was sprayed with INT. White spots indicate the presence of antibacterial activity.
RESULTS AND DISCUSSION
Isolation and identification of endophytic fungal isolates
Totally of 26 endophytic fungi were isolated from various parts of cinnamon plant, in which 12 isolates associated with cinnamon stem (C1BP1-C1BP12), and 14 isolates of endophytic fungi associated with cinnamon leaves (C1DP1-C1DP14).
Based on morphological characters of endophytic fungi, so they were classified as genus Pestalotiopsis (8 isolates), genus Xylaria (3 isolates), genus Colletotrichum (2 isolates), and genus Fusarium (1 isolate). Two isolates could only be identified at family level, ie Dematiaceae, and 10 isolates only identified at class level which is Coleomycetes. A complete morphological characters of endophytic fungi associated with cinnamon present in Table 1, and macroscopic picture of some isolates was in Figure 1.
Table 1. Morphological characters of endophytic fungi isolated from cinnamon plant (C. burmannii)
No.
|
Isolate
|
Genus/Family/
Class
|
Morphological Characters
|
1
|
C1BP-1
|
Pestalotiopsis
|
White creamish cottony mycelia; black perithecia; some produce brownish exudates
|
2
|
C1BP-2
|
Pestalotiopsis
|
White creamish cottony mycelia; black perithecia; some produce brownish exudates
|
3
|
CIBP-3
|
Xylaria
|
White cottony mycelia
|
4
|
C1BP-5
|
Coelomycetes
|
White creamish hairy mycelia
|
5
|
C1BP-6
|
Dematiaceae
|
Dark grey and black cottony mycelia
|
6
|
C1BP-7
|
Coelomycetes
|
White creamish hairy mycelia
|
7
|
C1BP-8
|
Dematiaceae
|
Dark grey and black cottony mycelia
|
8
|
C1BP-9
|
Coelomycetes
|
White creamish hairy mycelia
|
9
|
C1BP-10
|
Pestalotiopsis
|
White creamish cottony mycelia; black perithecia; some produce brownish exudates
|
10
|
C1BP-11
|
Pestalotiopsis
|
White creamish cottony mycelia; black perithecia; some produce brownish exudates
|
11
|
C1BP-12
|
Xylaria
|
Creamish brown and grey mycelia
|
12
|
C1BP-13
|
Coelomycetes
|
Grey creamish yellow green hairy mycelia, produce brown exudates
|
13
|
C1DP-1
|
Coelomycetes
|
White creamish hairy mycelia
|
14
|
C1DP-2
|
Xylaria
|
Creamish brown and grey mycelia
|
15
|
C1DP-3
|
Pestalotiopsis
|
White creamish cottony mycelia; black perithecia; some produce brownish exudates
|
16
|
C1DP-4
|
Coelomycetes
|
White creamish hairy mycelia
|
17
|
C1DP-5
|
Fusarium
|
White creamish hairy mycelia
|
18
|
C1DP-6
|
Colletotrichum
|
White and dark grey hairy mycelia, orange perithecia
|
19
|
C1DP-7
|
Coelomycetes
|
White creamish hairy mycelia
|
20
|
C1DP-8
|
Pestalotiopsis
|
White creamish cottony mycelia; black perithecia; some produce brownish exudates
|
21
|
C1DP-9
|
Pestalotiopsis
|
White creamish cottony mycelia; black perithecia; some produce brownish exudates
|
22
|
C1DP-10
|
Pestalotiopsis
|
White creamish cottony mycelia; black perithecia; some produce brownish exudates
|
23
|
C1DP-11
|
Coelomycetes
|
White creamish hairy mycelia
|
24
|
C1DP-12
|
Colletotrichum
|
White and dark grey hairy mycelia, orange perithecia
|
25
|
C1DP-13
|
Coelomycetes
|
White creamish hairy mycelia
|
26
|
C1DP-14
|
Coelomycetes
|
Grey creamish yellow green hairy mycelia, produce brown exudates
|
Figure 1. Single colonies of several endophytic fungi isolated from cinnamon stem and leaves. (Note.: Pestalotiopsis: 1, 2, 8, and 9. Colletotrichum: 6, and 7. Coelomycetes: 4 and 10. Dematiaceae: 5. Xylaria sp: 3)
Ten isolates could be identified morphologically (macroscopic) only to the class level, ie Coelomycetes (10 isolates). This is due to fungal isolates usually did not have specific organ by the time of observation. The formation of specific organ of fungal isolates can be stimulated by cultivating fungal isolate on its host plant. This has been done on endophytic fungus Diaporthe sp. that associate with the tea plant.[18]
Endophytic fungi that could be identified to the genus level and most commonly found was Pestalotiopsis (8 isolates). Pestalotiopsis usually found as plant pathogen[19], but in the last decade this genus has been isolated as endophytic fungi of several plant species. At least 20 species of Pestalotiopsis have been reported from 2000 to 2010 as endophytic fungi from 21 plant species[19], and this is the first time for Pestalotiopsis isolated from Cinnamomum burmannii.
Antibacterial screening of endophytic fungal extract
Endophytic fungi were cultivated in liquid medium (GYP and PDB), then extracted with ethyl acetate. The weight of concentrated extracts was shown in Table 2 and the bioautogram of the extracts was shown in Fig. 2.
Fig. 2. Bioautogram of endophytic fungal extracts isolated from cinnamon (C. burmannii )
plant against S. aureus InaCC-B4 (left) and E.coli InaCC-B5 (right). White area
against a purple background indicated antibacterial activity
Table 2. The diameter of inhibition zone (DIZ) of endophytic fungal extracts isolated
from cinnamon plant against S.aureus dan E.coli by TLC-bioautography
No
|
Fungal Endophyte
|
GYP
|
PDB
|
Extract
weight (mg)
|
DIZ (mm)
|
Extract weight (mg)
|
DIZ (mm)
|
S. aureus
|
E. coli
|
S. aureus
|
E. coli
|
1
|
Pestalotiopsis sp. C1BP-1
|
45.4
|
5.0
|
6.5
|
60.7
|
16.0
|
20.0
|
2
|
Pestalotiopsis sp. C1BP-2
|
30.2
|
-
|
5.5
|
20.8
|
8.0
|
12.0
|
3
|
Xylaria sp. C1BP-3
|
28.7
|
-
|
5.5
|
20.1
|
9.5
|
9.0
|
4
|
Coelomycetes C1BP-5
|
7.8
|
5.0
|
8.0
|
9.0
|
-
|
6.0
|
5
|
Dematiaceae C1BP-6
|
10.8
|
10.0
|
12.0
|
11.3
|
-
|
9.0
|
6
|
Coelomycetes C1BP-7*
|
-
|
-
|
-
|
-
|
-
|
-
|
7
|
Dematiaceae C1BP-8
|
-
|
nt
|
nt
|
12.8
|
6.5
|
11.5
|
8
|
Coelomycetes C1BP-9
|
105.3
|
11.0
|
6.5
|
166.40
|
9.0
|
6.0
|
9
|
Pestalotiopsis C1BP-10
|
234.7
|
-
|
-
|
32.3
|
-
|
-
|
10
|
Pestalotiopsis C1BP-11
|
-
|
nt
|
nt
|
30.6
|
-
|
-
|
11
|
Xylaria C1BP-12
|
37.6
|
-
|
-
|
40.2
|
-
|
-
|
12
|
Coelomycetes C1BP-13
|
30.2
|
-
|
-
|
15.6
|
-
|
13.0
|
13
|
Coelomycetes C1DP-1
|
-
|
8.0
|
-
|
12.5
|
6.0
|
5.0
|
14
|
Xylaria C1DP-2
|
28.7
|
-
|
8.0
|
15.9
|
8.0
|
4.0
|
15
|
Pestalotiopsis C1DP-3
|
-
|
nt
|
nt
|
18.8
|
5.0
|
-
|
16
|
Coelomycetes C1DP-4
|
11.4
|
-
|
5.5
|
8.6
|
15.0
|
20.0
|
17
|
Coelomycetes C1DP-5
|
7.8
|
-
|
-
|
8.7
|
-
|
-
|
18
|
Fusarium C1DP-6
|
92.8
|
-
|
-
|
70.4
|
6.0
|
6.5
|
19
|
Colletotrichum C1DP-7
|
9.5
|
-
|
-
|
7.6
|
-
|
-
|
20
|
Pestalotiopsis C1DP-8
|
29.6
|
-
|
-
|
15.0
|
-
|
-
|
21
|
Pestalotiopsis C1DP-9
|
14.2
|
-
|
-
|
18.2
|
-
|
-
|
22
|
Pestalotiopsis C1DP-10
|
14.0
|
6.0
|
6.5
|
15.1
|
-
|
4.5
|
23
|
Coelomycetes C1DP-11
|
-
|
nt
|
nt
|
43.4
|
-
|
6.0
|
24
|
Colletotrichum C1DP-12
|
31.6
|
-
|
-
|
30.0
|
5.0
|
-
|
25
|
Coelomycetes C1DP-13
|
10.9
|
-
|
-
|
5.4
|
-
|
-
|
26
|
Coelomycetes C1DP-14
|
17.8
|
-
|
-
|
59.1
|
-
|
4.0
|
Note : GYP : Glucose Yeast Pepton , PDB : Potato Dextrose Broth, nt : no tested, E.c : E. coli, S.a : S.aureus * Coelomycetes C1BP-7 : no growth on both culture media (GYP and PDB)
The result of antibacterial assay of endophytic fungal extract by bioautography showed that some extracts have the capability to inhibit the growth of S. aureus or E. coli (Figure 2). This is indicated by the formation of white area around the extracts. There are 18 endophytic fungal extracts that could inhibit the growth of S. aureus with the diameter of inhibition zone (DIZ) were in the range of 5.00 -16.00 mm, while the number of active extracts that inhibit the growth of E. coli are 24 extracts with the DIZ were in the range of 4.00 - 28.00 mm[Fig.2]. Two endophytic fungi (Pestalotiopsis sp. C1BP-01 and Pestalotiopsis sp. C1BP-02) have the ability to produce metabolites with potent antibacterial activity against S. aureus and E. coli when cultivated on PDB medium (Table 2). Several bioactive compounds isolated from the endophytic fungi Pestalotiopsis have been reported, such as taxol as anticancer drug produced by P. versicolor[20], P. breviseta[21], P. microspora[22], jesteron and hydroxy jesteron as antifungal produced by P. jesteri[23], while pestasin[24] and isopestasin[25] are produced by P. microspora as antifungal and antioxidant.
Three endophytic fungi isolated from cinnamon plants were identified to the genus Xylaria (CIBP-3, C1BP-12 and C1DP-2). Endophytic fungi belonging to the genus Xylaria have been widely reported in some plant species, such as Cinchona ledgeriana[26], Toona sinensis[27], Albertisia papuana[28], and Gingko biloba.[29] Several bioactive metabolites have also been reported to be produced by fungi of the genus Xylaria in the laboratory, such as 7-amino-4-metilkumarin[29], phloroglucinol[28], cytochalasin H and cytochalasin H2[30], cytochalasin Z27, cytochalasin Z28, cytochalasin Z18, cytochalasin E and secco-cytochalasin E.[27] The extract of Xylaria sp. C1BP-03 cultured on PDB medium showed strong antibacterial activity with the DIZ of 9.5 and 9.0 mm against S. aureus and E. coli, respectively. But, the extract of Xylaria sp. C1DP-03 is more potent against S. aureus (DIZ = 8.0 mm) than E.coli (DIZ = 4.0 mm)
Two endophytic fungi isolated from cinnamon plants were identified as genus Colletotrichum, but only Colletotrichum sp. C1DP-12 extract which has inhibitory effect against the growth of S. aureus (DIZ = 5.0 mm).
Endophytic fungi belonging to the genus Colletotrichum is generally known as plant pathogens. In the last two decades, there were many endophytic fungi of the genus Colletotrichum have been isolated.[31] Endophytic fungus Colletotrichum gloeosporioides isolated from the pepper plant (Piper nigrum) has the capability in producing piperina[32] as typical chemical compound of its host plants.
Fungi of the genus Fusarium also known as plant pathogens against many types of plants, especially banana plant with its Fusarium wilt disease.[33] Fungal infections caused by genus Fusarium are often followed by mycotoxin production, in which mycotoxin is one of the major problems in food ingredients derived from plants such as corn. The role of Fusarium as endophytes in several plants have been well described[34]
Fig. 3. Bio-autographyc screening of active antimicrobial compounds of endophytic
fungal extracts isolated from cinnamon plant, mobile phase : dichloromethane :
methanol (10 :1). White bands of the separated compounds indicated antibacterial
activity against S.aureus InaCC-B4 (left) and E.coli InaCC- B5 (right)
The active extract were then eluted with a mobile phase of dichoromethane : methanol (10: 1) in order to separate the chemical compounds of the extract. Figure 3. shows that the chemical compounds of endophytic fungal extracts isolated from cinnamon plant were more active in inhibiting the growth of E. coli bacteria. This is indicated by the white areas of E. coli were wider than that of S.aureus.
The result of TLC-bioautography showed that all chemical compounds of the extracts with retention factor (Rf) up to 0.8 have the inhibitory effect on the growth of S.aureus. The growth inhibitory effect on E.coli occurs a long the path of endophytic fungal extracts chromatogram. This indicates that the relatively more polar chemical compounds in both extract affects only the growth of S. aureus, while the non-polar and polar chemical compounds inhibit the growth of E. coli .
CONCLUSION
The results of this study showed that endophytic fungi isolated from cinnamon plants (Cinnamomum burmannii) produce bioactive metabolites which have antibacterial properties. Overall, the results of initial testing of endophytic fungi extracts isolated from cinnamon plants showed stronger activity against E. coli than S. aureus. However, there is possibility that pure compounds of these extracts will be more sensitive in inhibiting the growth of S. aureus. Therefore, further research is needed to determine the active compounds of endophytic fungal extracts isolated from cinnamon plant.
ACKNOWLEDGEMENTS
The research was funded by Kompetitif Research Project of Indonesian Institute of Sciences 2013-2015 and IFS Grant with contract number F-4613-2.
REFERENCES
[1] Higginbotham SJ, AE. Arnold, A Iban˜ ez, C. Spadafora, PD. Coley. (2013)
Bioactivity of fungal endophytes as a function of endophyte taxonomy and the
taxonomy and distribution of their host plants. PLoS ONE 8(9): e73192.
[2] Liang H., Y Xing., J.Chen, D.Zhang, S.Guo, and C.Wang (2012). Antimicrobial
activities of endophytic fungi isolated from Ophiopogon japonicus (Liliaceae).
BMC Complementary and Alternative Medicine 12:238
[3] Rodriguez, RJ., JF.White Jr, AE. Arnold, and RS. Redman (2009). Fungal
Endophytes : Diversity and Functional Roles. New Phytologist 182 : 314-330.
[4] Zhao J., T. Shan, Y. Mou, L. Zhou (2011). Plant derived bioactive compounds
produced by endophytic fungi. Mini Rev Med Chem 11(2):159-68.
[5] Newman, DJ. and GM. Cragg (2007). Natural products as sources of new drugs
over the last 25 years. Journal of Natural Products 70 : 461-477.
[6] Owen, NL. and N. Hundley (2004). Endophytes—the chemical synthesizers inside
plants. Science Progress, vol. 87( 2) : 79–99.
[7] Strobel, GA., and B. Daisy (2003). Bioprospecting for microbial endophytes and
their natural products. Microbiology and Molecular Biology Reviews 67 : 491-502.
[8] Kharwar RN, SK.Verma, A. Mishra, SK. Gond, VK. Sharma, T. Afreen, A. Kumar
(2011). Assessment of diversity, distribution and antibacterial activity of endophtic
fungi isolated from a medicinal plant Adenocalymma alliaceum Miers. Symbiosis.
55(1):39-46
[9] Gimenez C., R. Cabrera, M. Reina, A. Gonzales-Coloma (2007). Fungal
endophytes and their role in plant protection. Current Organic Chemistry 11(8) :
707-720.
[10] Ezra D, WM.Hess, and GA.Strobel (2004). New endophytic isolates of Muscodor
albus, a volatile-antibiotic-producing fungus. Microbiology 150: 4023–4031
[11] Santiago C., C. Fitchett, MHG .Munro, J. Jalil, , and J. Santhanam (2012). Cytotoxic and antifungal activities of 5-Hydroxyramulosin, a compound produced by an endophytic fungus isolated from Cinnamomum mollisimum. Evidence-Based Complementary and Alternative Medicine volume 2012 , Article ID 689310, 6 pages. http://dx.doi.org/10.1155/2012/689310
[12] Agusta A, S. Maehara, K. Ohashi, P. Simanjuntak and H. Shibuya (2005).
Stereoselective oxidation at C-4 of flavans by the endophytic fungus Diaporthe sp.
isolated from a tea plant. Chem.Pharm. Bul., 53 (12), 1565-1569.
[13] Barnett, H.L. and B.B. Hunter (1998). Illustrated genera of imperfect fungi. 4th ed.
USA: Prentice-Hall, Inc.
[14] Ellis, M.B. (1971). Dematiaceous hyphomycetes. England: Commonwealth
Mycological Institute.
[15] Domsch, K.H., W. Gams, and T.H. Anderson (1980). Compendium of soil fungi.
Vol 1. London: Academic Press.
[16] Sutton, B.C.(1980). The Coelomycetes. England: Commonwealth Mycological
Institute.
[17] Webster, J. (1980). Introduction to fungi. 2nd ed. Melbourne: Cambridge University Press.
[18] Agusta, A. (2006). Bioproduksi (+)-epiepoksidon oleh jamur endofit Diaporthe sp.
E yang diisolasi dari tanaman teh. Berita Biologi, 8(3), 209-214.
[19] Maharachchikumbura SSN., G. Liang-Dong, E. Chukeatirote, AH.Bahkali, and KD. Hyde (2011). Pestalotiopsis- morphology, phylogeny, biochemistry and diversity. Fungal Diversity 50: 167-187.
[20] Kumaran RS., HJ. Kim, and H. Byung-Ki. (2011). Taxol-producing fungal endophyte, Pestalotiopsis species isolated from Taxus cuspidata. Journal of Bioscience and Bioengineering vol. 111(6): 731(20)
[21] Kathivaran G. and SV. Raman (2010). In vitro taxol production, by Pestalotiopsis
breviseta - A first report. Fitoterapia vol. 81(6): 557-564.
[22] Li JY., RS. Sidhu, A. Bollon, and GA. Strobel (1998). Stimulation of taxol
production in liquid cultures of Pestalotiopsis microspora. Mycological Research
vol. 102 (4) : 461-464.
[23] Li JY. and GA. Strobel (2001). Jesterone and hydroxy-jesterone antioomycete
cyclohexenone epoxides from the endophytic fungus Pestalotiopsis jester. Phytochemistry vol. 57(2) : 261–265
[24] Harper JK, AM.Arif, EJ. Ford, GA. Strobel, JA. Porco Jr., DP. Tomer, KL. Oneill, EM. Heider, and DM. Grant (2003). Pestacin: a 1,3-dihydro isobenzofuran from Pestalotiopsis microspora possessing antioxidant and antimycotic activities. Tetrahedron vol. 59 (14) : 2471–2476
[25] Strobel G., E. Ford, J. Worapong, JK. Harper, AM. Arif, DM. Grant, PCW.Fung, and RMW. Chau (2002). Isopestacin, an isobenzofuranone from Pestalotiopsis microspora, possessing antifungal and antioxidant activities. Phytochemistry vol 60 (2) : 179–183
[26] Shibuya H., C. Kitamura, S. Maehara, M. Nagahata, H. Winarno, P. Simanjuntak, HS. Kim, Y. Wataya, and K. Ohashia (2003). Transformation of Cinchona Alkaloids into 1-N-Oxide Derivatives by Endophytic Xylaria sp. Isolated from Cinchona pubescens. Pharm. K. Bull. 51(1): 71—74
[27] Zhang Q., J. Xiao, QQ. Sun , JC. Qin, G. Pescitelli, and JM. Gao (2014).
Characterization of cytochalasins from the endophytic Xylaria sp. And their
biological functions. J. Agric. Food Chem 62 (45): 10962–10969
[28] Fathoni A, M. Ilyas, Praptiwi, AH. Cahyana, and A. Agusta ( 2013). Skrining dan
isolasi metabolit aktif antibakteri dari kultur jamur endofit dari tumbuhan
Albertisia papuana Becc. Berita Biologi, 12(30), 307-314.
[29] Liu X., M. Dong, X. Chen, M. Jiang, X. Lv, J. Zhou ( 2008). Antimicrobial
activity of an endophytic Xylaria sp.YX-28 and identification of its antimicrobial
compound 7-amino-4-methylcoumarin. Appl Microbiol Biotechnol (2008) 78:241-
247
[30] Li Y, Lu C., Huang Y., Li Y., and Shen Y.( 2012). Cytochalasin H2, a new cytochalasin, isolated from the endophytic fungus Xylaria sp. A23. Rec. Nat. Prod. 6 (2) :121-126.
[31] Manamgoda DS., D. Udayanga, L. Cai, E. Chukeatirote, KD. Hyde (2013). Endophytic Colletotrichum from tropical grasses with a new species C. endophytica. Fungal Diversity vol. 61 (1) : 107-115.
[32] Chithra S., B. Jasim, P. Sachidanandan, M. Jyothis, EK. Radhakrishnan (2014).
Piperine production by endophytic fungus Colletotrichum gloeosporioides isolated
from Piper nigrum. Phytomedicine 21:534–540.
[33] Garcia-Bastidas F., N. Ordóñez, J. Konkol, M. Al-Qasim, Z. Naser, M. Abdelwali, N. Salem, C. Waalwijk, RC. Ploetz, and GHJ. Kema 2014. First Report of Fusarium oxysporum f. sp. cubense Tropical Race 4 Associated with Panama Disease of Banana outside Southeast Asia. Plant Disease vol. 98 (5) : 694
[34] Bacon, CW and Yates, IE. 2006. Endophytic Root Colonization by Fusarium species: Histology,Plant Interactions, and Toxicity. In : Soil Biology, Volume 9 Microbial Root Endophytes. B. Schulz, C. Boyle, T. N. Sieber (Eds.) Springer-Verlag Berlin Heidelberg. http://link.springer.com/chapter/10.1007%2F3-540-33526-9_8#page-1
Dostları ilə paylaş: |