Proceeding s of Th e 5th Annu al I nt er nat ional Conf er ence Syiah Kuala Universit y ( AI C Unsyiah) 2 01 5
I n conj unct ion w it h The 8th I n t ern at ion al Con feren ce of Ch em ical Eng ineering o n Science and Applicat ions ( Ch ESA) 2 0 15 Sept em ber 9 - 1 1, 2 01 5, Banda Aceh, I ndon esia
Com post Oil Pa lm a n d I n dige n ou s En doph y t ic Fu n gi Effe ct
on Ba sa l St e m Rot in Oil Pa lm Se e dlin g
1*
Lisnawit a,
2Ham idah Hanum , and
3Ahm ad Rafiqi Tant awi
1,2
Program St udy of Agroecot echnology, Facult y of Agricult ure, Universit y of Sum at era
Ut ara, Padang Bulan, Medan 20155, I ndonesia;
3
Program St udy of Agrot echnology, Facult y of Agricult ure, Universit y of Medan Area, Medan
20223, I ndonesia
* Corresponding Aut hor
: it am uis@yahoo.com
Abst r a ct
Basal st em rot ( BSR) caused by Ganoderm a is an im port ant disease of oil palm in Nort h Sum at ra, I ndonesia. Cont rol of BSR is com plex , because t he disease occur s in t he dy nam ic env ironm ent at t he int erface of t he root s w it h soil. Com post w it h endophy t ic fungi as a biological fert ilizer offer possible adv ant ages and event ually suppressed ganoderm a grow t h. A st udy using t w o com post oil palm ( em pt y fruit bunch and palm oil m idrib- leaf com post ) cont aining indigenous endophy t ic fungi
(Trich oderm a koningii, T. v iride, and Aspergillus sp.) singly and in a m ix t ure w as conduct ed t o
det erm ine pot ent ial of t hem t o cont r ol Ganoderm a. The result show ed t hat 16 w eek s aft er art ificial inoculat ion, all t reat m ent s aren't significant difference reduced BSR incidence but com post cont aining endophy t ic fungi hav e pot ent ial t o inhibit t he pat hogen show ed BSR incidence in a m ix t ure t reat ed low er t han singly t reat ed seedlings.
Ke y w or ds:Aspergillu s sp., com post , Ganoderm a, oil palm , Trichoder m a koningii, T. v iride
I n t r od u ct ion
Oil palm (Elaeis guineensis) is one of t he largest plantat ion com m odit y indust ries in I ndonesia. Current ly, I ndonesia is one of t he world’s largest producer and exporter of oil palm . Areas of oil palm have increased from 135.000 hectares in 1970 t o 8.91 m illion in 2011 ( Pusat Data dan Sistem I nform asi Pertanian, 2013) , which producing an average palm oil yield approxim at ely 31 t onnes in 2014 ( ht tp: / / www.indonesia- invest m ents.com , 2015) . One of province wit h the second largest oil palm plant ed area in I ndonesia is Nort h Sum at ra wit h oil palm planted area is 1.2 m illion hectares ( Direkt orat Jenderal Perkebunan, 2012) .
One m aj or problem in oil palm is basal stem rot ( BSR) disease, caused by Ganoderm a spp., which is a serious t hreat t o t he oil palm indust ry in Sout h East Asian count ries, especially Malaysia and I ndonesia. The disease is let hal and in t he past few decades has been spreading rapidly. The disease spreads t hrough root t o root cont act. During plant ing any lit t le piece of rot t ing t runk is rem oved as t his could be a pot ent ial source of Ganoderm a in t he current stand (Hasan and Turner, 1998) . The disease progresses slowly, but event ually all infect ed plants becom e dead. This is because t he disease cannot be detect ed at t he early st ages.
Field cont rol of BSR by cont act chem icals have not been very successful ( Soepena et al., 2000) even wit h t hose in vit ro screened effect ive against t he fungus ( Khairudin, 1990; Teh, 1996) . The current int erest in reducing t he applicat ion of chem ical fert ilizer and increasing dem and for com bined effect s of beneficial fungi and organic com post can have great im pact on crop product ion and sust ainable agricult ure. Com post wit h endophyt ic fungi as a biological fert ilizer offer possible advantages and event ually suppressed ganoderm a growt h. A st udy using t wo com post oil palm ( em pt y fruit bunch and palm oil m idrib- leaf com post ) cont aining indigenous endophyt ic fungi (Trichoderm a koningii, T. viride, and Aspergillus sp.) singly and in a m ixt ure was conduct ed t o determ ine potent ial of t hem t o cont rol Ganoderm a.
M a t e r ia ls a nd M e t h ods
Pr ep a r a t ion of Ga node r m a in ocu lu m on ru bb er w ood b lock s
Fresh rubber wood blocks (12 cm x 6 cm x 6 cm , weighing approxim at ely 450 g) were washed wit h
water and autoclaved for 20 m in at 121oC. One block each was put in heat - resist ant polypropylene bags ( 15 cm x 33 cm x 0.05 m m t hick m at erial) and 100 m L of potat o dext rose agar ( PDA) added as supplem entary nut rient for Ganoderm a. Since inoculat ion of the blocks with Ganoderm a plugs can int roduce cont am inat ion, t he bags were closed by drawing its open end t hrough a 4 cm diam eter polyvinyl chloride ( PVC) t ubing 2 cm long and t he rem aining hole plugged wit h cot ton. The bags wit h
rubber wood blocks and PDA were aut oclaved at 121oC for 30 m in. After sterilizat ion and cooling, t he
Proceeding s of Th e 5th Annu al I nt er nat ional Conf er ence Syiah Kuala Universit y ( AI C Unsyiah) 2 01 5
I n conj unct ion w it h The 8th I n t ern at ion al Con feren ce of Ch em ical Eng ineering o n Science and Applicat ions ( Ch ESA) 2 0 15 Sept em ber 9 - 1 1, 2 01 5, Banda Aceh, I ndon esia
rubber wood block in t he bag was rot ated t o ensure t hat it was well covered wit h t he agar before t he lat t er solidified. When the agar had solidified, five pieces five day- old Ganoderm a cult ure t aken wit h cork borer and inoculat ed on each rubber wood block. The inoculated blocks in t he bags were t hen incubated in a dark cabinet at 28 ± 2oC for t hree weeks unt il fully colonized by Ganoderm a m ycelium
Pr ep a r a t ion of e nd op hy t ic fun gi
Trichoderm a koningii, Trichoderm a viride, and Aspergillus sp. were isolated from t he oil palm roots in Gunung Melayu, Asahan, Nort h Sum at ra and cultured on PDA m edium . All of endophyt fungi were prepared using rice m edium , t hen form ulated wit h zeolit e powder ( 1: 9 / v: v) .
Ap plica t ion of com post oil p a lm con t a in ing in dige n ous e nd op hy t ic fu ng i
Experim ent s carried out in t he glasshouse at Facult y of Agricult ure, Universit y of Sum at ra Utara, Medan, I ndonesia, using a fact orial random ized block design. A tot al of 72 healt hy oil palm seedlings t hree m ont hs old were used in t his experim ent. 300 g of com post single or com binat ion wit h 30 g of endophyt ic fungi inoculat ed before or after art ificial inoculat ion of Ganoderm a on oil palm seedlings.
Effe ct of com p ost oil p alm a nd ind ig e n ou s e nd op hy t ic fun gi on BSR in cid e n ce
At t he end of t he experim ent ( 4 m ont hs/ 16 weeks) , t he oil palm seedlings were split longit udinally t o observe for root and st em decay and t o visually assess t he severit y of t he internal sym pt om s based on t he proport ion of root and bole t issues dam aged by Ganoderm a. The est im at ion was based on t he following m odified scale ( Abdullah et al., 2003; I lias 2000) . 0 = Healt hy plants, 1 = Appearance of whit e fungal m ass on any part of plants, wit h or wit hout chlorot ic leaves, 2 = Appearance of basidiom a on any part of plant s wit h chlorot ic leaves ( 1–3 leaves) , 3 = Form at ion of basidiom a on any part of plant s with chlorot ic leaves ( > 3 leaves) , 4 = Form at ion of well- developed basidiom a and t he plants dried. Disease severit y ( DS) for t he int ernal sym pt om s was calculated by t he form ula of Liu et al. ( 1995) as under. DS
( int er nal) = ( num ber of seedlings in t he rat ing x rat ing num ber) / ( tot al num ber of
seedlings assessed x highest rat ing) x 100.The disease incidence and disease severit y percent ages were t ransform ed by arcsine and analyzed using ANOVA wit h t he m eans com pared by the LSD ( P ≤ 0.05) .
Resu lts a nd D iscussion
Art ificial infect ion of plants by contact wit h the inoculum block carrying Ganoderm a is an effect ive st rategy for inducing the infect ion. The assessm ent of disease severit y was based on t he sequent ial progression of disease using index values 0- 4. Lower disease severit y indicates t he effect iveness of t he t reatm ent s in suppressing Ganoderm a BSR in oil palm t hrough in vivo assessm ent . I n general, 16 weeks after art ificial inoculat ion, all t reat m ent s aren't significant difference reduced BSR incidence but com post containing endophyt ic fungi have potent ial to inhibit t he pathogen showed BSR incidence in a m ixt ure t reat ed lower than singly t reated seedlings ( Table 1) . This st udy showed t hat com post containing endophyt ic fungus (T. koningii, T. viride, and Aspergillus sp.) have a good potent ial as biological agent s t o inhibit Ganoderm a.
Table.1. Disease severit y of oil palm plant s at BSR 16 weeks after art ificial inoculat ion
Tr e a t m e n ts D ise ase se ve r ity ( % )
K0W1E0 33.3
K0W1E1 33.3
K0W1E2 25.0
K0W1E3 25.0
K0W2E0 25.0
K0W2E1 25.0
K0W2E2 25.0
K0W2E3 25.0
K1W1E0 25.0
K1W1E1 8.3
K1W1E2 16.7
K1W1E3 16.7
K1W2E0 25.0
K1W2E1 16.7
K1W2E2 16.7
Proceeding s of Th e 5th Annu al I nt er nat ional Conf er ence Syiah Kuala Universit y ( AI C Unsyiah) 2 01 5
I n conj unct ion w it h The 8th I n t ern at ion al Con feren ce of Ch em ical Eng ineering o n Science and Applicat ions ( Ch ESA) 2 0 15 Sept em ber 9 - 1 1, 2 01 5, Banda Aceh, I ndon esia
K1W2E3 8.3
K2W1E0 25.0
K2W1E1 16.7
K2W1E2 16.7
K2W1E3 8.3
K2W2E0 25.0
K2W2E1 16.7
K2W2E2 8.3
K2W2E3 16.7
Not e : K0 : wit hout com post ,
K1 : em pt y fruit bunch com post , K2 : palm oil m idrib- leaf com post ,
W1 : inoculat ion Ganoderm a before endophyt ic fungi W2 : inoculat ion Ganoderm a aft er endophyt ic fungi E0 : wit hoout endpohyt ic fungi
E1 : T. viride E2 : T. koningii E3 : Aspergillus sp.
The com posit ion of t he m icroorganism s in com posts is affect ed by t he chem ist ry of t he m aterials from which t he com post is prepared ( Cast año et al., 2011) . Com post s wit h high lignocellulosic substances ( t ree barks) are m ost ly colonized by Trichoderm a spp. I n cont rast , grape pom ace, wit h low cellulosic substances and high sugars, becom es colonized by Penicillium spp. and Aspergillus spp. ( Kut t er et al. 1983; Gorodecki and Hadar 1990) . I n com posts, in addit ion t o t he well- docum ented effect s of m icrobial populat ions against several soil- borne diseases, induct ion of resistance has also been report ed as an addit ional biocont rol m echanism against both foliar and root diseases ( Zhang et al. 1996, 1998; Kavroulakis et al. 2005; Nt ougias et al., 2008) . Reduct ion in disease severit y was dem onst rat ed against Sept oria lycopersici in t om at o ( Kavroulakis et al., 2005) , bact erial leaf spot in radish caused by Xant hom onas cam perst ris pv. Arm oraciae ( Krause et al., 2003) , ant hracnose in cucum ber caused by Collet ot richum orbiculare (Zhang et al., 1996) and Bot ryt is cinerea in cucum ber and m elon ( Segarra et al. 2007; Yogev et al., 2010) . Moreover com post water ext ract root t reat m ent induce system ic resist ance t o ant hracnose caused by C. orbiculare in cucum ber and C. coccodes in pepper ( Sang and Kim 2011) .
Figure 1. Signs and sym pt om s of BSR, caused by Ganoderm a; A, B. necrot ic lesion on infected plant ( arrow) , C. The leaves of infected plant s were chlorot ic ( a) , and healt hy leaves (b)
The ideal of biocont rol st rat egy at t em pt s t o int roduce or prom ote the act ivit y of biocont rol agents only when and where t hey are needed or are m ost effect ive, and m inim ises wasteful applicat ion of inoculum on non- t arget habitat s. Different m echanism s have been suggest ed as being responsible for t he effect s of biocont rol agents; t hey include com pet it ion for space and nut rient s, secret ion of chit inolyt ic enzym es, m ycoparasit ism and product ion of an inhibit ory com pound (Haram et al., 1996; Zim and et al., 1996). Thus, the t im ing of when t o apply t he t reatm ent s is im port ant due t o t he
( a )
A
B
C
( b)
Proceeding s of Th e 5th Annu al I nt er nat ional Conf er ence Syiah Kuala Universit y ( AI C Unsyiah) 2 01 5 part of the plants, then form ed basidiom a before t he plants dried (Figure 1C) .
Con clusion
Am ount of 16 weeks after art ificial inoculat ion, all t reat m ents aren't significant difference reduced BSR incidence but com post (em pt y fruit bunch and palm oil m idrib- leaf com post ) containing indigenous endophyt ic fungi (Trichoderm a koningii, T. viride, and Aspergillus sp.) have pot ent ial t o inhibit the pathogen showed BSR incidence in a m ixt ure t reat ed lower t han singly t reated seedlings.
Ack n ow led g em e n t
This research was supported by KEMENRI STEK DI KTI t hrough The Nat ional Com pet it ive Grant s Schem e MP3EI ( 2014- 2015). Many t hanks for t he financial support.
