Nghien ciPU - Ky thuat
Tai lieu tham khao 1. Awasthi R K., Dixit S. C. (2009), "Chemical composition of Curcuma longa leaves and rhizome oil from the plains of Northern India", J. Young Pharm., 1940.312-316.
2. Kyong-Up Baik, Sang-Hun Jung and Byung-Zun Ahn (1993), "Recognition of pharmacophere of ar- turmerone for its anticancer activity". Archives ofPhar.
Research, Volume 16, Numbers, pp 254-256.
3. N. Y. Qin, F Q. Yang, Y. T. Wang, S. P. Li (2007),
"Quantitative determinaton of eight components in rhizome (Jianghuang) and tuberous root (Yujin) of Curcuma longa using pressurized liquid extraction and gas chromatography-mass spectrometry", Joumal of Pharm. and Biomedical Analysis. 43, pp 486-492.
4. Pranee LertsutOiiwong, Khanittha Noomun, Nutthapon Jongaroonngamsang, Pomchai Rojsitthisak, Ubonthip Nimmannit (2008), "Preparation of alginate nanocapsules containing turmenc oil", CadDohydrale Polymers, 74, pp 209-214.
(Ngay nhdn bdi: 12/05/2015 - Ngdy duydt ddng: 04/08/2015)
Khao sat dieu kien nuoi cay chung vi nam noi sinh phan lap tir cay buoi
{Citrus maxima (Burm.) Merr.)
Vo Thi Ngoc My'', Nguyen Van Thanh' Nguyin Dinh NgaS Nguyen Thi Thuy Van^
' Tru&ng Dai hoc Nguyen Tdt Thanii - Dgi hoc Y-Dirac Tp. Ho Chi Mmh 'E-mail: [email protected]
Summary
TWo species of endophytic fungi Pseuderotium and Epicoccum were identified from the stems of Citrus maxima (Burm.) Men. Some strains of Epicoccum and Pseuderotium were isolated from Citrus maxima (Burm.) Merr. and determined capable of producing secondary metabolites of microcidal activity a by disc diffusion. Besides, an appropriate composition and favorable condition of culture were established for high productivity of the metabolites against Staphylococcus aureus. Streptococcus fecalis, Pseudomonas and MRSA: Carbon source - 4% glucose; nitrogen sources - 20% potatoes; and incubation - at room temperature for seven days. Preliminary experiments proved ethyl acetate rather suitable for extraction of the antibacterial compounds.
K e y w o r d s : Citrus maxima. Epicoccum, Pseuderotium, Endophyte, Endophytic fungi.
Dat van de
Theo t h i n g ke eua To chuc Y t l t h l gidi (WHO) cd khoang 80% dan so t h l gidi, chu y l u tLP cac nudc phdt trien dua vdo cdc loai t h u l c truyen t h i n g (cd nguon goc tCr thuc vat) de cham sdc sue khoe. Trong 119 loai hop ehdt hda hpc, it nhat 90 logi ed nguon g l c t d thuc vat. Day la cdc logi thuoc dang d u p c s u dung ngdy cang nhilu d n h i l u q u i c gia. Van de ddt ra la eac chat ed hoat tfnh sinh hpc trong edy Id do ehinh eay sinh ra hay la k i t qua eua m l i lien he tuang sinh vdi cac VI sinh vgt ngi sinh co ich trong md thuc vat.
Vi sinh vdt ndi sinh thye vdt Id nhung vi sinh vat
phat trien ben trong cae md sau cua cay nhung thu'dng khong gdy benh cho cdy chu •''. Trong do, vi khuan va vi nam la cac vi sinh vat ndi sinh thudng ggp nhat. T i n s6 phdn lap d u p c vi sinh vgt ndi sinh Id vi nam c h i l m ty le cao hon, vi vgy CO hpi tim ra ehung vi nam n5i sinh mdi tir cdy Id rat cao. NhOng nghien cuu gdn day cua Hawksworth va Rossman nam 1997 da u d c tinh cd d i n mpt trigu loai vi nam khae nhau, nhung ehi khoang 100.000 loai da d u p c md ta P". Hign tgi va t u a n g lai, vi nam ngi sinh se !a nguon da dgng sinh hgc doi dao, mdi Ig trong dd ed nhilu lodi chua d u p e b i l t d i n .
TAP CHI Dir<?(C HQC - 8/2015 (SO 472 .NAM 55)
• Nghien ctru - Ky thuat
Vi nam ndi sinh ed mdi quan he chat ehe vdi cdy ehu, s o dung cdc ehat dinh d u d n g trong edy de ton tai, mang lai n h i l u lpi ich cho cay nhu tao ra ede san pham trao doi ehdt co hoat tinh sinh hgc nhu eac hormon sinh trudng, cac c h i t khdng sinh cd kha nang bao vg cdy khoi ede vi sinh vgt gdy benh. Nhieu tdi ligu cho thay khi cgng sinh trong mo thye vgt, vi nam ndi sinh da san sinh ra nhieu chat cd hogt tinh sinh hgc nhu khang k h u i n va khang nam, vi dy nhu Kennedia nigriscans, mgt loai cdy leo d Gc da d u p c dan gian s u dyng sap nhya de tri v l t thuang, sat trung; ti> cdy nay da phdn lgp d u p c chung Streptomyces sp. NRRL 30562 mdi san xuat khdng sinh munumbicins p h i rdng i^'. Cf quy mo rpng, vi ndm ndi sinh cd tdc dpng quan trpng trong ndng nghigp, rung vd dang d u p c quan tdm nghign ci>u va tilm ndng cCia chiing cd y nghTa rat ldn trong cac ITnh v y c y hgc, ndng nghigp vd cdng nghigp i^'i.
Hg Rutaceae d Vidt Nam da dgng, co khodng 140 loai, trong dd cam, chanh, quyt, b u d i ... ed n h i l u ung dung trong d u p e pham, thyc pham...
Nhieu cong trinh nghign cuu cho thay tinh dau cua vo qua ede edy hg Rutaceae co tdc dung an t h i n , chiia cam ciim. T u nhiJng ddc tinh tren cho thay, hp Rutaceae chua hg vi n l m ndi sinh phong phu, tiet n h i l u chat khang khuan, khdng nam vd cd 'if nghTa trong vi^c d i l u trj bgnh i"".
D l gop phan khai thac tilm nang cac edy hp Rutaceae iam t h u l c phyc vy cdng tac d i l u tri, chung toi tien hdnh nghien cuu khao sdt d i l u ki$n nu6i c l y chCing vi n l m ndi sinh phan ldp tip cay budi (Citrus maxima (Burm.) Merr.).
Vat lieu va phu'O'ng phap nghien CU'U
Vat lieu
Cac bp phan nhu than, canh, la, hoa, vd qua cua cay budi - Citrus maxima (Burm.) Merr. thu tai v u d n nhd Vung Tau. D l i t u p n g thtj hoat tinh bao gom: Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 29213, Streptococcus faecalis ATCC 29212, MRSA ATCC 43300.
Phu'ong phdp nghien cu'u
Xi> ly mau va phdn l$p vi nam noi sinh Chpn cde cdy budi song tnang moi trudng t y nhign, khde mgnh, khdng cdi cpc, khdng bj
sau hai hay bieu hign bgnh nhu ddm la, vdng eay... Mau cay thu thgp d u p c rua saeh vdi nudc, d l rao noi khd mat, sau dd d u p e XLP ly tai phdng thi nghigm trong dieu kign vd trung. Ivliu dupe dat trgn mdi trudng thgch nudc, khong chua cac ehat dinh d u d n g khae. Theo doi s y phat trien eua vi ndm ndi sinh tir mdu cay trong thdi gian khoang 5 - 1 4 ngay. Sau dd, cay c h u y i n sang mdi trudng P D A e d bo sung chloramphenicol 0 , 1 % (ue c h l s y tang tru'dng cua vi khuan ndi sinh), u d nhidt do phong vd theo doi khoang 7 - 14 ngay hoac lau han tuy theo tde do phdt trien cua vi ndm npi sinh phan lgp dupe. T i l p tuc cay c h u y i n vi nam ndi sinh sang moi trudng chua c h i t dinh dudng (PDA) roi u d nhiet dp phong va theo ddi s y phat t r i l n , xdc djnh dgc diem vi nam vd khao sat tinh khang khuan '^'.
Sang Igc vi nam ngi sinh cho chat bien dwang khang khuan
Vi n l m ndi sinh nuoi trgn mdi trudng P D A t u 5 - 1 4 ngay, d nhigt dp phong d u p c t h u kha nang khang cac chung vi khuan £ coli. Staphylococcus aureus, MRSA, Streptococcus faecalis bang phuang phap khulch tan t u khoanh thach thO.
Khao sat anh huxmg cua thai gian va thdnh phan moi trtecyng nuoi cay anh hipong den si^
san sinh hoat chat khang khuan cua vi nam ndi sinh
Tien hdnh nuoi cdy cac chung vi n l m npi sinh CO hogt tinh khang khuan d nhigt dd 25-30 °C va lupng bao t u ban dau 10^ CFU/mi tren cac dTa petn ehua 20 ml moi trudng ran vdi nguon nito vd nguon earbon d u p c thay doi d ede nong dp khdc nhau' Pepton casein 1 % ; dich khoai tdy:
200 g khoai tdy/1000 ml nude eat i^.
Ojnh tinh nhanh bdng phuang phap k h u l c h tan qua dTa giay khang sinh: Cac dich ehilt cua vi nam ngi sinh t u moi trudng nuoi cdy rdn sau khi cd cho bay hoi, cdn dupc hoa trong ethanol hay dung mSi de bay hoi. Sau dd t i m tryc tiep 1 lupng xac dinh djch cdn da d u p c hda trong dung moi d l bay hoi len dfa giay khdng sinh, d l kh6 t y nhien. Sau khi d u l l sach dung moi, dat dia giay len mgt thgch da trai vi khuan thO nghigm, u 37 °C trong 24 gid. Do dudng kinh vdng khang khuan.
T i l n hanh nuoi c l y vi n l m npi sinh da phdn lap tren mdi trudng eho hogt tinh khdng khuan cao nhat da d u p c khao sat. Sau dd c h i l t hpp chat ed hogt tinh khang khuan cua vi n l m ndi sinh.
TAP CHl DUpc HQC - 8/2015 (SO 472 NAM 55)
• Nghien CLPU - Ky thuat
Djnh danh nhung chung vi nam ngi sinh CO hoat tinh khang khuin
Djnh danh so bg d u d i kinh h i l n vi sau khi nhupm vdi Laetophenol cotton blue d l so bp phdn logi nam C l y nam tren m6i tm'dng chuyen bigt d l dinh danh (PDA). C l y trgn hop dTa petri d l do dudng kinh, quan sdt mdu sac va theo doi hinh dgng khuan lac. Cay tren lam de quan sdt hinh t h l , do dat kieh thudc.
Ket qua
Phdn lap v i nam noi s i n h tip cay bLPoi T u 10 Chung phan lap dupe tir cay budi
thi ed 3 ehung cho tde dpng khang khuan, trong dd gom ed 1 chung eo tac dpng khang khuan ylu va 2 chung cd tde dpng vira.
Sang Ipc vi nam noi s i n h c d hoat tinh khdng khuan
Cae chung vi n l m ngi sinh sau 7 ngay nuoi eay tren mdi trudng PDA d u p e khao sat tdc dgng khang khuan tren £. coli vd S. aureus, S. faecalis va MRSA thi co 3 chung cho hogt tinh khdng khuan, trong do: Chung BC2 va BT2 d i u cho hoat tinh khang d E. coli, S. aureus, S. faecalis va MRSA. Chung BT3 tac dpng y l u trgn E. coli va S. aureus.
K^
khdng E. coli khang S. aureus khang S. faecalis khang MRSA Hinh 1 : ChCing BC2 vd BT2 khang cac chung vi khuin: E. coli, S. aureus, S. faecalis, MRSA Khao sat thdnh phan m o i trLPO'ng va t h d i
gian nuoi cay anh hu'dng den su> sdn sinh hogt chat khang khuan cua vi nam ndi sinh
Nguon nita Id khoai tdy vd nguon carbon la glucose
Bang 1 : Hieu qua san smh chdt khdng khuin cua chung BC2
T u moi trudng c o ban la mdi trudng PDA, vi nam npi sinh dupc nudi cay tren moi ti-udng vdi nguin nito Id khoai tdy vd nguon carbon la glucose (nong dp glucose thay doi t u 2%, 3%, 4%) d nhifit dp phong, lupng bao t i i ban ddu 10^ CFU/ml.
TTioi gian nuoi cay (ngay)
Dipong kinh vong khang khuan cua chung BC2 (mm) E. coli S. faecalis
5 7 9 11
12 14 14 12
12 14 13 12
15 18 17 15
13 11 0
13 11 0
15 15 12
14 12 10 Bang 2: Hieu qua san sinh chdt khdng khuan cCia chOng BT2 Thai gian nuoi cay
(ngay)
Duxmg kinh vbng khang khuin cua chung BT2 (mm)
S. aureus E. coli S faecalis ivlRSA 2% 3% 4% 2% 3% 4% 2% 3% 4% 2% 3%
11 15 12 11
14 18 17 12
11 13 11 9
11 13 11 9
12 15 15 12
11 14 12 10
11 12 13 10
14 17
n
12
T^P CHi DUgtC Hpc - 8/2015 (S6 472 NAM 55)
Nghien cu'u - Ky thuat
Hoat tinh khdng khuin d ea 2 chung BC2 va BT2 d i u dat d muc cao nhat d moi tiudng ed nong dp glucose 4%, nuoi eay d nhigt dp phong d ngdy thu 7.
Nguon nita la khoai tay va nguon carbon Id saccharose
Bang 3: Hieu qua san sinh chat khdng khuin cua chung BC2
Cac Chung vi nam noi sinh d u p c nuoi cay tren moi trudng vdi ngudn nito Id khoai tdy va ngudn carbon la saccharose (nong dp saccharose thay doi tLP 0,5%, 1 % , 2%) d nhigt dp phong, lupng bao t u ban d i u 10^ CFU/mi.
Thai gian nuoi cay (ngay)
Dirang kinh vong khang khuin cua chung BC2
S. aureus E. coli S. faecalis MRSA 0,5% 1% 2% 0,5% 1% 2% 0.5% 1% 2% 0.5%, 1%
14 14 14
10
«
12 014 16 15 M
0 13 11 0
0 14 11 10
15 15 15 12
11 14 12 10
11 12 13 9 Bdng 4: Hieu qui san sinh chat khdng khuan cua chung BT2 Thai gian nuo
(ngay) 5 7 9
cay S
0,5%
0 13 13
aureus 1%
0 13 12
2%
15 17 16
Suong kinh vong khang khuan ciia cliijng BT2
E coll S faecalis MRSA 0,5% 1% 2% 0,5% 1% 2% 0,5% 1% 2%
14 13 13 13
0 15 15 12
0 14 12 0
0 15 13 9
14 16 15 12 Hogt tinh khang khuan d ca 2 chung BC2 va
BT2 deu dgt muc eao n h i t d moi trudng cd nong dp saccharose 2%, nuoi cay d nhigt dg phdng d ngay thu 7, nhung hoat tinh khang k h u i n kgm hon so vdi khi nuoi cay d moi trudng cb nong dg glucose 4%.
Nguon nitty la pepton casein va natri nitrat trong moi truwig Czapeck-Dox
Khi nudi cay BC2 vd BT2 trong mdi trudng Sabouraud (nguon nito la pepton, nguon carbon la glucose); mbi trudng Czapeck-Dox (nguon nito la natri nitrat, n g u i n carbon la sucrose) ca 2 chung vi nam BC2 vd BT2 hodn todn khdng san smh hogt chat khang k h u i n .
Khdo sdt dnh hud'ng cua thdi gian nudi cay tren su> san sinh hp'p chat khdng khuan cua chiing BC2 va BT2
Bang 5: Hoat tinh khdng khuan cua chat
Nubi cay chung vi nam npi sinh tren mdi trudng c6 nguon nito la khoai tay, nguon carbon la glucose 4%. Hogt ehat khang khuan dtrpc san sinh bdi BC2 va BT2 b i l n thien theo thai gian nuoi cay, d ngdy t h u 7 higu qua san sinh hogt chdt khdng E coli, S. aureus, S. faecalis va MRSA la ldn nhat cho vbng khang cao nhat so vdi eae ngay edn lai.
Nuoi cay va chiet tach hp'p c h i t khdng khuan tip cdc chung BC2 va BT2
T i l n hanh ehilt hogt chat khang khuan tCp cac dfa petri da d u p c nudi c l y cde chung BC2 va BT2 da d u o c khao sat (moi trudng vdi nguon nito- khoai tay; nguon carbon: glucose 4%) vdi ethanol 96%, sau khi boc hoi h i t dung mbi, edn se d u p e chiet xuat vdi ethanol. Sau do lac phan b l vdi cdc dung mbi cb dp phdn c y c tang dan.
chidt tCr mdi truang nuoi cay chung BC2 Duvmg kinh vong khang khuin trung binh cua chung BC2 (mm)
£ coli S. aureus S. faecalis I^RSA Chi4t xuat bang ethanol
Ctiiet xuSt blng n-hexan Cflietxuif blng ethylacetat Chi^t xult bang ethanol 10%
16 0 25 15
14 0 25 16
13 0 18 11
17 0 20 16
TAP CHl DUpC HQC - 8/2015 (SO 472 NAM 55)
• Nghien CIFU - Ky thuat
Hoat chdt cd tinh khang khuan phdn b l chu y l u trong chdt ehilt vdi ethyl acetat, tuy nhien dich c h i l t e u l i eung (ethanol 10%) vdn edn eho
Bang 6: Hoat tinh khdng khuin cua chdt chiet tO moi truang nuoi cay chung BT2
heat tinh khang khuan t u a n g doi cao trgn S.
aureus, M R S A v a £ co/; (bang 5).
Dumig kinh E.adi
vong khang khuin bvng binh cda chung BC2 (mm) S. aureus S. Recalls MRSA Chiet xult blng ethanol
Chiet xult blng hexan Chilt xuat bang ethyl acetat Chiet xuat bang ethanol 10%
17 0 27 14
17 0 25 11 Hoat c h i t cd tinh khang khuin phan b l chu y l u trong chdt chiet vdi ethyl acetat, tuy nhien vdl djch c h i l t c u i i cung (ethanol 10%) thi hoat chat khdng khuan van cbn tuong doi eao d 2 Chung vi khuan S. faecalis va MRSA (bang 6).
Cd t h l hoat chat khdng khuan trong c h i l t xuat ethanol 10% ehua d u p c phan b l hodn toan trong ethyl acetat hoac c6 2 chat eung cho hogt tinh nhung cd dp phdn c y c khae nhau.
Djnh danh nhirng chung vi n l m npi sinh c6 hoat tinh khdng k h u i n
Chung B T2 mang ddc didm cua Epicoccum sp.
Dae diem khdm n l m tren moi trudng PDA: Too dp phdt t r i l n nhanh 0 = 5 cm/ 7 ngay. Khdm nlm trbn, mdu vang kem, spi nam mdu trdng mjn, vien ngoai mau trang, thanh n h i l u vbng sac to thay doi t u trong ra ngoai, sinh sdc to vang nau khulch tan ra mbi trudng. Mat trdi mdu vang s i m gan ndu.
BT2/ PDA (MP) BT2/ PDA (MT) Hinh 2: Dae diem hinh Dgc diem khi quan sat bdng kinh hien vi: Spi nam cb vdch ngan, khbng mau khi non nhung cho mdu nau khi trudng thanh. Cuong sinh bao t u ngdn. Bdo tCr t r i n sinh ra don dbc tren cdc gid sinh bdo t u tran khbng rb rgt, tgo thanh tirng ddm d i n g thdi hay don ddc, cd day rbng. Bdo tCp nhom trong eae cym nhb gpn (khli bao tu). Bdo tLP mdu ndu, vach thd, ed hoae khbng cd vbng.
K i l u phat sinh bao t u tran: nay c h l i .
Bao tLP BT2 Cuong sinh bdo tCp BT2 thai cOa chung BT2
Chung BC2 mang dae diem cua Pseuderotium sp.
Ddc d i l m khdm nam trdn mdi trudng PDA:
Toe dp phdt t r i l n nhanh 0) = 6,5 cm/ 6 ngay.
Khdm nam spi bdng ban dau khdng mdu sau dd c h u y i n sang mau xdm tro, ngoai ria co dgng spi, spi nam mjn, sinh sde to xanh ra ngoai mbi trudng. Mgt trai ban dau mdu xanh den sau phdt t r i l n chuyen sang mdu den.
BC2/PDA (MP) BC2/PDA (MT) Bdo ti> BC2 rieng le Hinh 3: Ddc diem hinh thai cija chung BC2
Bao li> BC2 dinh chum
TAP CHI Dl/gfC HQC - 8/2015 (SO 472 NAM 55)
Nghien cu'u - Ky thuat
Dge d i l m khi quan sdt bdng kinh hien vi: T h l qua hinh c l u den bau due, khbng cb l5. Bdo t u sinh san v d tinh giong n h u Sporothrix sp. vdi eubng sinh bao tCp dai, vdch tron, khbng mau, dinh thon nhpn. Cac c u i n g sinh bao ti> hop thanh vdi nhau 1 gbc 30°. T l bao sinh bao tCr hinh b i u due, dinh mpt chum bao tCp khbng eubng d ngpn.
Bao l u hinh bau dye d i n hoi eau, vach tron.
Ban ludn
Nhung vi nam npi sinh sinh chat b i l n dudng cd hoat tinh rat da dgng ve hinh thai ldn each sinh san va hau nhu chCing la nhij-ng vi ndm cb hinh thue sinh sdn dge bigt, Cf nghien euu nay, 2 chiing da dupe phdn lgp BC2 va BT2 d u p c nudi c l y trgn mbi trudng ed thdnh phan: khoai tdy 2%, glucose 4 % va agar 1,5% thi hoat tinh sinh ra mgnh n h i t d ngdy t h u 7. Phdn ldn cac tai lieu lien quan d i n qua trinh e h i l t tdeh c h i t b i l n d u d n g eho tdc dgng khdng khuan t u mdi trudng nudi eay vi nam npi sinh d i u eho thay ethyl acetate la dung mbi thich hpp. T u o n g t y trong khao sat chCing toi s u dung ethanol d l c h i l t chat khang k h u i n tir mbi trudng nudi eay vi nam nbi sinh sau db ide phan b l Ibng - Ibng vdi cac dung mdi cb dp phdn eye khdc nhau, k i t qua cho thay chat khdng khuan d u p c phdn bb trong ethyl acetat la chu y l u .
Ket luan
Phdn lgp va sang Ipc d u p c 3 chung vi nam npi sinh d eay budi, tip db xde djnh d u p c 2 chung vi nam BC2 va BT2 cb tdc dpng khdng khuan trdn E. coli, S. aureus, S. faecalis va MRSA. Khao sdt mbi trudng vd thdi gian nubi cay thieh hpp de 2 Chung BC2 vd BT2 san sinh c h i t b i l n dudng khang khuan tot nhat: Dich khoai tay (khoai tdy/
n u d c cat: 20 g/l); glucose 4%; agar 1,5%, eho hoat tinh khdng khuan mgnh nhat d ngay thCp 7. Budc dau khdo sdt quy trinh e h i l t taeh hpp c h i t khdng k h u i n tCr mdi trudng rdn nubi eay 2 chung vi nam npi sinh BC2 va BT2, xae dinh d u p e dung mbi thieh h p p de ehilt thb hpp chat khang khuan. Bang phuang phap djnh danh d y a vao hinh thdi vi n l m , b u d c dau ghi nhgn
2 Chung vi nam nbi sinh (vi nam) cho hoat chat b i l n dudng khang khuan mgnh phdn lgp t u b u d i thupc ehl Pseuderotium (chung BC2) va thupe chi Epicoccum (ehung BT2).
Loi cam an: Chung tdi xin chan thanh cam o n Bp mdn Vi sinh - ki sinh, Khoa Dupc, Dai hge Y-Dupe Tp. Hb Chi Minh; Khoa Dupc, Trudng Dgi hpc Nguygn T i t Thanh da hb trp va tgo d i l u kien d l chung tbi hoan thdnh nghien c u u ndy.
Tai lieu tham khao 1. Strobel G , Daisy B. (2003), "Bioprospecting for microbial and their nature products", Microbiology and Moiecular Biology Reviews, pp. 491-502.
2. Kumud Chandra Kandpal, D. A Jain, Umesh Kumar, Rashmi Tnpathi and T Siva Kumar (2012),
"Isolation and screening of endophytic actinomycetes producing antibacterial compound from Citrus aurantifolia Fruit", European Joumal of Experimental B/o/ogy, 2, 1733-1737.
3.MaksumRadji,AtiekSumiati,RenJtaRachmayani and Bema Elya (2011), "Isolation of fungal from Garcinia mangostana and their antibacterial activity", Afiican Journal of Biotechnology, 10,103-107.
4. Min-Yuan Ho, Wen-Chuan Chung, Hung-Chang Huang, Wen-Hsia Chung and Wen-Hsin Chung (2012),
"Identification of endophytic fungi of medicinal herbs of Lauraceae and Rutaceae with antimicrobial property", Taiwania, 57, 229-241.
5. Pham Quang Hung and K. Annapurna (2004),
"Isolation and characterization of endophytic bacteria in Soybean {Glycine, sp)", Omonrice,12, 92-101.
6. R. E. Jalgaonwala, B. V. Mohite and R.T. Mahajan (2010), "Evaluation of endophytesfortheir antimicrobial activity from Indigenous medicinal plants belonging to North Maharashtra region India", Intemational Journal on Pharmaceutical and Biomedical Research, 1, 136- 141,
7. R. X. Tan and W. X. Zou (2001), "Endophytic fungi a rich source of functional metabolites", Nat Prod. Rep., 18,448-459
8. U. F. Castillo (2000), "Munumbicins, wide spectrums antibiotics produced by Streptomyces NRRL 30562, endophytic on Kennedia nigriscans".
Microbiology. 148. 2675- 2685.
(Ngay nhdn bdi: 11/02/2015 - Ngdy duydt dang: 04/08/2015)
TAP CHI DUgfC HpC - 8/2015 (SO 472 NAM 55)
