Colonization Ability of Biological Control Agent Tricoderma spp on Cocoa Pod and Seedling | Sriwati | Proceedings of The Annual International Conference, Syiah Kuala University - Life Sciences & Engineering Chapter 5844 11970 1 PB
Proceedings of The 5 t h Annual I nt ernat ional Conference Syiah Kuala Universit y ( AI C Unsyiah) 2015
I n conj unct ion wit h The 8 t h I nt ernat ional Conference of Chem ical Engineering on Science and Applicat ions ( ChESA) 2015
Sept em ber 9- 11, 2015, Banda Aceh, I ndonesia
Coloniza t ion Abilit y of Biologica l Cont r ol Age nt Tr icode r m a spp on
Cocoa Pod a nd Se e dling
1
* Rina Sriwat i, 1 Tj ut Kham zurni, 2 Elvira I skandar, and 1 Mit ra Tri Annisa Lest ari
1
Agro t echnology Depart m ent , Facult y of Agricult ure, Syiah Kuala Universit y, Banda Aceh
I ndonesia
2
Agribusiness Depart m ent , Facult y of Agricult ure, Syiah Kuala Universit y, Banda Aceh
I ndonesia
* Corresponding Aut hor : [email protected]
Abst r a ct
Som e of Trichoderm a species as ant agonist ic fungi are usually considered soil m icroorganism ,
They colonize plant root s, som e- t im es form ing a sym biot ic relat ionship. Three species of
Trichoderm a ( T. virens, T. harzianum and T. asperellum ) have been inoculat ed on cacao seedling
and cocoa pod. Trichoderm a species can be re- isolat ed from surface st erilized cacao seedling,
including t he st em and leaf, root , and pod t hen observed their colonizat ion abilit y. Fungal hyphae
were observed under t he m icroscope em erging from t he leaf, st eam , root of seedling and pod as
soon as 1 day aft er t heir isolat ion from surface st erilized cacao seedling and pod. All Trichoderm a
species were able to ent er and m ake colonizat ion. The highest percent age of colonizat ion
occurred in t he T. harzianum by 73.3% ( leaves) , 46.7% ( t runk) and 86.7% ( root s) . While
colonizat ion on t he skin cocoa pod ( epiderm is) also has a different percent age, t he highest
percent age indicat ed in t he t reat m ent of T. harzianum by 63.3% . We conclude t hat T. harzianum
bet t er biological cont rol agent base on t heir abilit y t o colonize all part of seedling and pod.
Trichoderm a species int o t he cacao st em , leaf, root and pod allowing syst em ic colonizat ion of t his
t issue.
Ke y W or d: Colonizat ion, Tricoderm a spp, cocoa pod, seedling, biological cont rol, challenge
I nt r oduct ion
Product ivit y of cocoa recent ly have been decreasing caused by pest and disease pat hogen. Som e
fungal pat hogens at t acked cacao plant s cause large yield losses. One of t he pat hogen, Phyt opht hora
Palm ivoracause black pod disease of cocoa rot . The high pot ent ial losses have been caused by t hi s
black pod disease. Biological cont rol m et hods t hat was effect ive and efficient wit h t he sust ainable
syst em .
Current ly m any bact erial and endophyt e fungi isolat ed from t he plant and root s. Those organism t hat
were ant agonist ic against soil borne pat hogens and pot ent ially could be use as biological cont rol
agent s. The use of biological agent s t o reduce t he num ber of inoculum pat hogen, reducing t he
pat hogen’s abilit y t o infect it s host and inhibit t he developm ent of pat hogens in t he host . Ant agonism
m echanism was t he abilit y of an organism t o inhibit t he growt h and developm ent of ot her organism s
( Cook & Baker, 1989) . Sriwat i et al., ( 2009) Det ect ed and I dent ified of Endophyt ic fungus
Trichoderm a associat ed in cacao plant in Aceh Sum at era. They report ed t hat t here are several species
of endophyt ic fungi associat ed in cacao leaves from East Aceh.
Trichoderm a is one ant agonist ic m icroorganism capable t o suppressing pat hogens and can be used as
a biocont rol against fungal pat hogens. Ut ilizat ion of t he fungus Trichoderm a also has pot ent ial as a
biological cont rol for ant agonism against several plant pat hogens, such as Fusarium sp, Rhizoct onia
solani and Phyt ium ( Ram ada, 2008) . Trichoderm a sp also t he saprophyt e m icroorganism s act ing as
organic decom poser.
They can degraded t he m at erials cont aining fibers, lignin and organic
com pounds, cont aining carbon and nit rogen used fr om organic m at t er ( Saraswat i & Sum arno, 2008) .
Trichoderm a abilit y t o produce ant ibiot ics and parasit e pat hogen in host plant t issue has been st udied
ext ensively ( Harm an et al., 2004) . There are sever al species of Trichoderm a are used as biocont rol
agent s include T. harzianum , T, Viridae, T. konigii, and T. virens ( Howell, 2003) . Muarif ( 2013)
report ed t hat T. virens w hich high concent rat ion wer e able t o inhibit P. palm ivora at t acks in cacao
seedling. I n addit ion, som e st rains of T. harzianum were able t o suppress t he developm ent of
Sclerot ium rolfsi on bean plant s by 30- 50% ( Papavizas & Lewis, 1989) . While research Rosm ana et
al., ( 2013) revealed t hat an effect ive and efficient way t o cont rol leaf blight disease on cacao seedlings
caused by soil- borne pat hogens by using T. asperellum . The use of T asperellum at a concent rat ion of
10 6 indicat es t he percent age and int ensit y of t he disease were lower t han t he cont rol t reat m ent .
203
Proceedings of The 5 t h Annual I nt ernat ional Conference Syiah Kuala Universit y ( AI C Unsyiah) 2015
I n conj unct ion wit h The 8 t h I nt ernat ional Conference of Chem ical Engineering on Science and Applicat ions ( ChESA) 2015
Sept em ber 9- 11, 2015, Banda Aceh, I ndonesia
One of t he problem in t he ut ilizat ion of Trichoderm a as biological cont rol agent s of plant diseases was
low colonizat ion abilit y ( Nurbailis & Mart in, 2009) . Research on t he effect iveness of som e species of
Trichoderm a as biological cont rol agent s have been carried out , but on t he abilit y of Trichoderm a spp
in colonized part s of t he plant has not been known. Therefore, it is necessary t o st udy t he abilit y of
endophyt es colonizat ion in som e Trichoderm a species on cacao seeds and pod.
M a t e r ia ls a nd m e t hods
This research was conduct ed at t he Laborat ory of Plant Pat hology Agrot echnology Depart m ent , Facult y
of Agricult ure Syiah Kuala Universit y. T. virens, T. harzianum , T. asperellum suspension were used for
inoculat ion t o seedling and pod. Com plet ely random ized design ( CRD) non fact orial consist ing of 3
t reat m ent s and 6 replicat ions have been done. Observat ion abst ain on cacao seeds and pod.
Table 1. Com posit ion of Som e Species of Trichoderm a Treat m ent
No
Pe r la k ua n
Ke t e r a nga n
1
Tv
T. virens concent rat ion 1 x 10 6
2
Th
T. har zianum concent rat ion 1 x 10 6
3
Ta
T. asperellum concent rat ion 1 x 10 6
Pure cult ures of T. virens, T. har zianum and T. asperellum has been grown on PDA m edia and
incubat ed for ± 2 weeks t o form spores. Cocoa seeds collect ed from cacao plant at ion Sare, Aceh
Besar dist rict . Cocoa seeds were germ inat ed on cot t on m edia t o det erm ine germ inat ion seed before
being t ransferred int o polybags. Seven days aft er germ inat ion cacao seedling were select ed t o obt ain
a good bean sprout s, t hen t ransferred t o soil in t he polybag m edia. Top soil t hat used has been sift ed
and st erilized using an aut oclave at 121 ° C for 30 m inut es. The st erilized soil m ixed wit h m anure in
t he rat io of soil and m anur e 2: 1 in polybag size of 1 kg. Each spore concent rat ion 1 x 10 6 of T. virens,
T. har zianum and T. asper ellum have been sprayed on seedling and pod.
Det erm ine t he colonizat ion of T. virens, T. harzianum and T. asperellum has been done by t aking t he
leaves, st em s and root s, sam pled wit h a diam et er of 1 cm by using cork borer, t he leaves, st em s and
root s were t aken 5 sect ions. These part were st erilized wit h NaClO ( sodium hypochlorit e) . All sam ples
were inoculat ed on PDA m edium for hypha observat ion.
Trichoderm a colonizat ion percent age in t he sam ples of seed and pod t hat have been isolat ed were
count ed and charact erized by m orphology charact erist ic of Trichoderm a growt h on t he PDA. Colonies
t hat grew ident ified m orphologically by Bar net t and Hunt er ( 1972) . Tr ichoderm a colonizat ion
percent age calculat ed using t he form ula:
Trichoderm acolonizat ion percent age =
x 100%
Re sult s
When leaf, st em , root and pod sect ions of cacao seedlings colonized by Trichoderm a, ht e hyphae were
observed wit hin 24 h aft er surface st erilizat ion. Trichoderm a hyphae appeared t o be em erging from
t he t ips of leaf, st eam and root ( Fig 1) and from pod ( Fig 2) . This was a general observat ion for m any
different
204
Percentage of Colonization (%)
Proceedings of The 5 t h Annual I nt ernat ional Conference Syiah Kuala Universit y ( AI C Unsyiah) 2015
I n conj unct ion wit h The 8 t h I nt ernat ional Conference of Chem ical Engineering on Science and Applicat ions ( ChESA) 2015
Sept em ber 9- 11, 2015, Banda Aceh, I ndonesia
c
c
60
50
d
b
d
40
c
30
a
20
c
leaf
b
b
a
a
stems
roots
10
0
Control
T. virens
T. harzianum
T. asperellum
Kind of Trichoderma
Figure 1. Pe r ce nt a ge of Coloniza t ion Tr ichode r m a on Ca ca o Se e dlings
Trichoderm a species was not lim it ed t o t he specific st rains of Trichoderm a shown. Trichoderm a were
isolat ed from colonized cacao seedlings aft er surface st erilizat ion t o verify t hat t hey were int ernally
colonized. Low percent age Trichoderm a was isolat ed from t he un- inoculat ed cont rol seedlings and pod
com pare wit h inoculat ed t reat m ent . All t hree Trichoderm a species st udied were re- isolat ed from leaf,
st eam and root . I nt erm ediat e, wit h em erging Trichoderm a hyphae t hen were observed and isolat ed.
Trichoderm a hyphae m ost oft en em erged from t he surface out side ( epiderm is) but were also observed
em erged from inside of pod. The percent age of isolat ing t he inoculat ed Trichoderm a st rain from
seedling wit h em erging hyphae was higher on T. harzanum and T virens st rains on t his st udied
com pere wit h T. asperellum and cont rol. The Trichoderm a isolat ed in highest percent age isolat ion of
Trichoderm a from each lat e sam ples were always consist ent from root seedling
Percentage of Trichoderma (% )
Tricoderm a species from pod em erging hyphae were always em erged in high percent age from for T.
harzianum . Out side skin of pod as epiderm is part was higher colonizat ion of Trichoderm a com pare
wit h inside pod ( endoderm is) ( Fig 2)
c
35
30
b
25
b
20
Epidermis
15
10
Endodermis
a
5
0
Kontrol
T. virens
T. harzianum T. asperellum
Kind of Trichoderma
Figure2. Pe r ce nt a ge of Coloniza t ion Tr ichode r m a on Ca ca o Pods
D iscussion
The high capabilit y of colonizat ion T. har zianum showed t hat T. harzianum m ost effect ive for use as
biological agent s. Alt hough Sriwat i at al 2015 report ed t hat T. virens Aceh isolat e has good pot ent ial
as biological cont rol for black pod disease on laborat ories experim ent but t he works of t his fung i have
m ore a less t han T. harzianum . Thrane et al., ( 2002) st at ed t hat T. harzianum produce enzym es such
as cellulase, glucanase and chit inase, could speed of growt h and ant agonist ic t o plant pat hogens.
Am ong som e Trichoderm a species have been ident ified, t he m ost com m on t ype used as biocont rol soil
borne pat hogen is T. harzianum ( Sunar wat i and Yoza, 2010) . The colonies of T. harzianum in t he root
syst em and it s consequences seen abilit y t o suppress t he growt h of soil borne fungal pat hogens ( Cook
in Ekowat i, 2000) . From part s of plant s t hat have been isolat ed, it is known t hat t he highest
205
Proceedings of The 5 t h Annual I nt ernat ional Conference Syiah Kuala Universit y ( AI C Unsyiah) 2015
I n conj unct ion wit h The 8 t h I nt ernat ional Conference of Chem ical Engineering on Science and Applicat ions ( ChESA) 2015
Sept em ber 9- 11, 2015, Banda Aceh, I ndonesia
percent age of colonizat ion found on t he root s of T. harzianum ( 56.53) and T. virens ( 56.53% ) . The
fungus T. harzianum and T. virens have t he sam e capabilit ies at t he root colonizat ion in t he 1 x 10 6 .
This was because t he rhizosphere around t he root t here are m any nut rient s t hat can be used as food
ingredient s for Trichoderm a growt h. T. harzianum capable of form ing colonizat ion of root s t hat can
ext end and im prove prim ary root growt h of t om at o plant s ( Herlina et al. 2004) . According t o Harm an
et al., ( 2004) , Trichoderm a spp as saprofit have good live in t he rhizosphere of plant s. I n addit ion,
Yedidia et al., ( 1999) also st at es t hat som e st rains of Trichoderm a able t o colonize and can be
endophyt ic in plant root t issues t hat causes increased act ivit y of t he com pound in t he resilience of t he
plant root s.
On t he out er skin of cocoa pod, t he m ost high percent age of colonizat ion indicat ed in t he t reat m ent of
T. har zianum by 31.3% , while t he inside part of cocoa pod showed no significant difference in each
t reat m ent . The highest percent age in in inside part in T. har zianum t reat m ent wit h an average
percent age of 26.24% . Nat urally, Trichoderm a present in t he cocoa fruit t issue wit h a very sm all
am ount . This is proven by t he Trichoderm a in t he cont rol t reat m ent an average of 5.8% . The addit ion
of Trichoderm a suspension at a cert ain concent rat ion can increase t he populat ion of Trichoderm a in
t he t issues of plant s and fruit . Sriwat i ( 2014) st at es t hat t he provision of Trichoderm a suspension at a
concent rat ion of 1 x 107 can suppress t he growt h of P. palm ivora cause fr uit rot disease of cocoa.
Conclusion
All Trichoderm a species were able t o ent er and m ake colonizat ion. The abilit y of t hird colonizat ion
Trichoderm a species on cacao seeds have different percent ages. Highest percent age of colonizat ion
occurred in t he T. harzianum . While colonizat ion on t he skin cocoa pod ( epiderm is) also has a different
percent age, t he highest percent age indicat ed in t he t reat m ent of T. har zianum . We conclude t hat T.
harzianum bet t er biological cont rol agent base on t heir abilit y t o colonize all part of seedling and pod.
Ack n ow le dge m e nt s
This work was support ed by a grant from t he Direct or General of hig her Educat ion, Depart m ent of
Nat ional Educat ion I ndonesia. We t hank t o Bryan Bailey at Laborat ory Sust ainable Perennial Crop Lab.
SPCL USDA- ARS Belt sville, MD, for fungal ident ificat ion work.
Re fe re n ce
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on cocoa seedlings. t hesis Unsyiah.
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developm ent of Fusarium wilt disease ( Fusarium oxysporum f. Sp. Cubense) . Journal Nat ur I ndonesia 13( 3) ,
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Proceedings of The 5 t h Annual I nt ernat ional Conference Syiah Kuala Universit y ( AI C Unsyiah) 2015
I n conj unct ion wit h The 8 t h I nt ernat ional Conference of Chem ical Engineering on Science and Applicat ions ( ChESA) 2015
Sept em ber 9- 11, 2015, Banda Aceh, I ndonesia
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207
I n conj unct ion wit h The 8 t h I nt ernat ional Conference of Chem ical Engineering on Science and Applicat ions ( ChESA) 2015
Sept em ber 9- 11, 2015, Banda Aceh, I ndonesia
Coloniza t ion Abilit y of Biologica l Cont r ol Age nt Tr icode r m a spp on
Cocoa Pod a nd Se e dling
1
* Rina Sriwat i, 1 Tj ut Kham zurni, 2 Elvira I skandar, and 1 Mit ra Tri Annisa Lest ari
1
Agro t echnology Depart m ent , Facult y of Agricult ure, Syiah Kuala Universit y, Banda Aceh
I ndonesia
2
Agribusiness Depart m ent , Facult y of Agricult ure, Syiah Kuala Universit y, Banda Aceh
I ndonesia
* Corresponding Aut hor : [email protected]
Abst r a ct
Som e of Trichoderm a species as ant agonist ic fungi are usually considered soil m icroorganism ,
They colonize plant root s, som e- t im es form ing a sym biot ic relat ionship. Three species of
Trichoderm a ( T. virens, T. harzianum and T. asperellum ) have been inoculat ed on cacao seedling
and cocoa pod. Trichoderm a species can be re- isolat ed from surface st erilized cacao seedling,
including t he st em and leaf, root , and pod t hen observed their colonizat ion abilit y. Fungal hyphae
were observed under t he m icroscope em erging from t he leaf, st eam , root of seedling and pod as
soon as 1 day aft er t heir isolat ion from surface st erilized cacao seedling and pod. All Trichoderm a
species were able to ent er and m ake colonizat ion. The highest percent age of colonizat ion
occurred in t he T. harzianum by 73.3% ( leaves) , 46.7% ( t runk) and 86.7% ( root s) . While
colonizat ion on t he skin cocoa pod ( epiderm is) also has a different percent age, t he highest
percent age indicat ed in t he t reat m ent of T. harzianum by 63.3% . We conclude t hat T. harzianum
bet t er biological cont rol agent base on t heir abilit y t o colonize all part of seedling and pod.
Trichoderm a species int o t he cacao st em , leaf, root and pod allowing syst em ic colonizat ion of t his
t issue.
Ke y W or d: Colonizat ion, Tricoderm a spp, cocoa pod, seedling, biological cont rol, challenge
I nt r oduct ion
Product ivit y of cocoa recent ly have been decreasing caused by pest and disease pat hogen. Som e
fungal pat hogens at t acked cacao plant s cause large yield losses. One of t he pat hogen, Phyt opht hora
Palm ivoracause black pod disease of cocoa rot . The high pot ent ial losses have been caused by t hi s
black pod disease. Biological cont rol m et hods t hat was effect ive and efficient wit h t he sust ainable
syst em .
Current ly m any bact erial and endophyt e fungi isolat ed from t he plant and root s. Those organism t hat
were ant agonist ic against soil borne pat hogens and pot ent ially could be use as biological cont rol
agent s. The use of biological agent s t o reduce t he num ber of inoculum pat hogen, reducing t he
pat hogen’s abilit y t o infect it s host and inhibit t he developm ent of pat hogens in t he host . Ant agonism
m echanism was t he abilit y of an organism t o inhibit t he growt h and developm ent of ot her organism s
( Cook & Baker, 1989) . Sriwat i et al., ( 2009) Det ect ed and I dent ified of Endophyt ic fungus
Trichoderm a associat ed in cacao plant in Aceh Sum at era. They report ed t hat t here are several species
of endophyt ic fungi associat ed in cacao leaves from East Aceh.
Trichoderm a is one ant agonist ic m icroorganism capable t o suppressing pat hogens and can be used as
a biocont rol against fungal pat hogens. Ut ilizat ion of t he fungus Trichoderm a also has pot ent ial as a
biological cont rol for ant agonism against several plant pat hogens, such as Fusarium sp, Rhizoct onia
solani and Phyt ium ( Ram ada, 2008) . Trichoderm a sp also t he saprophyt e m icroorganism s act ing as
organic decom poser.
They can degraded t he m at erials cont aining fibers, lignin and organic
com pounds, cont aining carbon and nit rogen used fr om organic m at t er ( Saraswat i & Sum arno, 2008) .
Trichoderm a abilit y t o produce ant ibiot ics and parasit e pat hogen in host plant t issue has been st udied
ext ensively ( Harm an et al., 2004) . There are sever al species of Trichoderm a are used as biocont rol
agent s include T. harzianum , T, Viridae, T. konigii, and T. virens ( Howell, 2003) . Muarif ( 2013)
report ed t hat T. virens w hich high concent rat ion wer e able t o inhibit P. palm ivora at t acks in cacao
seedling. I n addit ion, som e st rains of T. harzianum were able t o suppress t he developm ent of
Sclerot ium rolfsi on bean plant s by 30- 50% ( Papavizas & Lewis, 1989) . While research Rosm ana et
al., ( 2013) revealed t hat an effect ive and efficient way t o cont rol leaf blight disease on cacao seedlings
caused by soil- borne pat hogens by using T. asperellum . The use of T asperellum at a concent rat ion of
10 6 indicat es t he percent age and int ensit y of t he disease were lower t han t he cont rol t reat m ent .
203
Proceedings of The 5 t h Annual I nt ernat ional Conference Syiah Kuala Universit y ( AI C Unsyiah) 2015
I n conj unct ion wit h The 8 t h I nt ernat ional Conference of Chem ical Engineering on Science and Applicat ions ( ChESA) 2015
Sept em ber 9- 11, 2015, Banda Aceh, I ndonesia
One of t he problem in t he ut ilizat ion of Trichoderm a as biological cont rol agent s of plant diseases was
low colonizat ion abilit y ( Nurbailis & Mart in, 2009) . Research on t he effect iveness of som e species of
Trichoderm a as biological cont rol agent s have been carried out , but on t he abilit y of Trichoderm a spp
in colonized part s of t he plant has not been known. Therefore, it is necessary t o st udy t he abilit y of
endophyt es colonizat ion in som e Trichoderm a species on cacao seeds and pod.
M a t e r ia ls a nd m e t hods
This research was conduct ed at t he Laborat ory of Plant Pat hology Agrot echnology Depart m ent , Facult y
of Agricult ure Syiah Kuala Universit y. T. virens, T. harzianum , T. asperellum suspension were used for
inoculat ion t o seedling and pod. Com plet ely random ized design ( CRD) non fact orial consist ing of 3
t reat m ent s and 6 replicat ions have been done. Observat ion abst ain on cacao seeds and pod.
Table 1. Com posit ion of Som e Species of Trichoderm a Treat m ent
No
Pe r la k ua n
Ke t e r a nga n
1
Tv
T. virens concent rat ion 1 x 10 6
2
Th
T. har zianum concent rat ion 1 x 10 6
3
Ta
T. asperellum concent rat ion 1 x 10 6
Pure cult ures of T. virens, T. har zianum and T. asperellum has been grown on PDA m edia and
incubat ed for ± 2 weeks t o form spores. Cocoa seeds collect ed from cacao plant at ion Sare, Aceh
Besar dist rict . Cocoa seeds were germ inat ed on cot t on m edia t o det erm ine germ inat ion seed before
being t ransferred int o polybags. Seven days aft er germ inat ion cacao seedling were select ed t o obt ain
a good bean sprout s, t hen t ransferred t o soil in t he polybag m edia. Top soil t hat used has been sift ed
and st erilized using an aut oclave at 121 ° C for 30 m inut es. The st erilized soil m ixed wit h m anure in
t he rat io of soil and m anur e 2: 1 in polybag size of 1 kg. Each spore concent rat ion 1 x 10 6 of T. virens,
T. har zianum and T. asper ellum have been sprayed on seedling and pod.
Det erm ine t he colonizat ion of T. virens, T. harzianum and T. asperellum has been done by t aking t he
leaves, st em s and root s, sam pled wit h a diam et er of 1 cm by using cork borer, t he leaves, st em s and
root s were t aken 5 sect ions. These part were st erilized wit h NaClO ( sodium hypochlorit e) . All sam ples
were inoculat ed on PDA m edium for hypha observat ion.
Trichoderm a colonizat ion percent age in t he sam ples of seed and pod t hat have been isolat ed were
count ed and charact erized by m orphology charact erist ic of Trichoderm a growt h on t he PDA. Colonies
t hat grew ident ified m orphologically by Bar net t and Hunt er ( 1972) . Tr ichoderm a colonizat ion
percent age calculat ed using t he form ula:
Trichoderm acolonizat ion percent age =
x 100%
Re sult s
When leaf, st em , root and pod sect ions of cacao seedlings colonized by Trichoderm a, ht e hyphae were
observed wit hin 24 h aft er surface st erilizat ion. Trichoderm a hyphae appeared t o be em erging from
t he t ips of leaf, st eam and root ( Fig 1) and from pod ( Fig 2) . This was a general observat ion for m any
different
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Percentage of Colonization (%)
Proceedings of The 5 t h Annual I nt ernat ional Conference Syiah Kuala Universit y ( AI C Unsyiah) 2015
I n conj unct ion wit h The 8 t h I nt ernat ional Conference of Chem ical Engineering on Science and Applicat ions ( ChESA) 2015
Sept em ber 9- 11, 2015, Banda Aceh, I ndonesia
c
c
60
50
d
b
d
40
c
30
a
20
c
leaf
b
b
a
a
stems
roots
10
0
Control
T. virens
T. harzianum
T. asperellum
Kind of Trichoderma
Figure 1. Pe r ce nt a ge of Coloniza t ion Tr ichode r m a on Ca ca o Se e dlings
Trichoderm a species was not lim it ed t o t he specific st rains of Trichoderm a shown. Trichoderm a were
isolat ed from colonized cacao seedlings aft er surface st erilizat ion t o verify t hat t hey were int ernally
colonized. Low percent age Trichoderm a was isolat ed from t he un- inoculat ed cont rol seedlings and pod
com pare wit h inoculat ed t reat m ent . All t hree Trichoderm a species st udied were re- isolat ed from leaf,
st eam and root . I nt erm ediat e, wit h em erging Trichoderm a hyphae t hen were observed and isolat ed.
Trichoderm a hyphae m ost oft en em erged from t he surface out side ( epiderm is) but were also observed
em erged from inside of pod. The percent age of isolat ing t he inoculat ed Trichoderm a st rain from
seedling wit h em erging hyphae was higher on T. harzanum and T virens st rains on t his st udied
com pere wit h T. asperellum and cont rol. The Trichoderm a isolat ed in highest percent age isolat ion of
Trichoderm a from each lat e sam ples were always consist ent from root seedling
Percentage of Trichoderma (% )
Tricoderm a species from pod em erging hyphae were always em erged in high percent age from for T.
harzianum . Out side skin of pod as epiderm is part was higher colonizat ion of Trichoderm a com pare
wit h inside pod ( endoderm is) ( Fig 2)
c
35
30
b
25
b
20
Epidermis
15
10
Endodermis
a
5
0
Kontrol
T. virens
T. harzianum T. asperellum
Kind of Trichoderma
Figure2. Pe r ce nt a ge of Coloniza t ion Tr ichode r m a on Ca ca o Pods
D iscussion
The high capabilit y of colonizat ion T. har zianum showed t hat T. harzianum m ost effect ive for use as
biological agent s. Alt hough Sriwat i at al 2015 report ed t hat T. virens Aceh isolat e has good pot ent ial
as biological cont rol for black pod disease on laborat ories experim ent but t he works of t his fung i have
m ore a less t han T. harzianum . Thrane et al., ( 2002) st at ed t hat T. harzianum produce enzym es such
as cellulase, glucanase and chit inase, could speed of growt h and ant agonist ic t o plant pat hogens.
Am ong som e Trichoderm a species have been ident ified, t he m ost com m on t ype used as biocont rol soil
borne pat hogen is T. harzianum ( Sunar wat i and Yoza, 2010) . The colonies of T. harzianum in t he root
syst em and it s consequences seen abilit y t o suppress t he growt h of soil borne fungal pat hogens ( Cook
in Ekowat i, 2000) . From part s of plant s t hat have been isolat ed, it is known t hat t he highest
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Proceedings of The 5 t h Annual I nt ernat ional Conference Syiah Kuala Universit y ( AI C Unsyiah) 2015
I n conj unct ion wit h The 8 t h I nt ernat ional Conference of Chem ical Engineering on Science and Applicat ions ( ChESA) 2015
Sept em ber 9- 11, 2015, Banda Aceh, I ndonesia
percent age of colonizat ion found on t he root s of T. harzianum ( 56.53) and T. virens ( 56.53% ) . The
fungus T. harzianum and T. virens have t he sam e capabilit ies at t he root colonizat ion in t he 1 x 10 6 .
This was because t he rhizosphere around t he root t here are m any nut rient s t hat can be used as food
ingredient s for Trichoderm a growt h. T. harzianum capable of form ing colonizat ion of root s t hat can
ext end and im prove prim ary root growt h of t om at o plant s ( Herlina et al. 2004) . According t o Harm an
et al., ( 2004) , Trichoderm a spp as saprofit have good live in t he rhizosphere of plant s. I n addit ion,
Yedidia et al., ( 1999) also st at es t hat som e st rains of Trichoderm a able t o colonize and can be
endophyt ic in plant root t issues t hat causes increased act ivit y of t he com pound in t he resilience of t he
plant root s.
On t he out er skin of cocoa pod, t he m ost high percent age of colonizat ion indicat ed in t he t reat m ent of
T. har zianum by 31.3% , while t he inside part of cocoa pod showed no significant difference in each
t reat m ent . The highest percent age in in inside part in T. har zianum t reat m ent wit h an average
percent age of 26.24% . Nat urally, Trichoderm a present in t he cocoa fruit t issue wit h a very sm all
am ount . This is proven by t he Trichoderm a in t he cont rol t reat m ent an average of 5.8% . The addit ion
of Trichoderm a suspension at a cert ain concent rat ion can increase t he populat ion of Trichoderm a in
t he t issues of plant s and fruit . Sriwat i ( 2014) st at es t hat t he provision of Trichoderm a suspension at a
concent rat ion of 1 x 107 can suppress t he growt h of P. palm ivora cause fr uit rot disease of cocoa.
Conclusion
All Trichoderm a species were able t o ent er and m ake colonizat ion. The abilit y of t hird colonizat ion
Trichoderm a species on cacao seeds have different percent ages. Highest percent age of colonizat ion
occurred in t he T. harzianum . While colonizat ion on t he skin cocoa pod ( epiderm is) also has a different
percent age, t he highest percent age indicat ed in t he t reat m ent of T. har zianum . We conclude t hat T.
harzianum bet t er biological cont rol agent base on t heir abilit y t o colonize all part of seedling and pod.
Ack n ow le dge m e nt s
This work was support ed by a grant from t he Direct or General of hig her Educat ion, Depart m ent of
Nat ional Educat ion I ndonesia. We t hank t o Bryan Bailey at Laborat ory Sust ainable Perennial Crop Lab.
SPCL USDA- ARS Belt sville, MD, for fungal ident ificat ion work.
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