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PAPER A2 - Effect Filter Cover of Seedlings in Direct Inoculation Screening of
Uromycladium tepperianum
for
Falcataria moluccana
Disease Tolerant
Asri Insiana Putri
1
, Liliana Baskorowati
1
, Nurhidayati
1
, Toni Herawan
1
, Siti Husna Nurrohmah
1 1
Center of Forest Biotechnology and Tree Improvement Research, Jl. Palagan Tentara Pelajar Km. 15, Purwobinangun, Pakem, Sleman,Yogyakarta, Indonesia, 55582,
Corresponding E-Mail: asriipyahoo.co.id
ABSTRACT
Effectively of direct inoculation screening of F. moluccana to gall rust disease tolerance in nursery needs to be proved in un-cultured pathogen like U. tepperianum. This technique
requires accurate simulation of natural environmental conditions for disease infection to host target. The aim of this study is to observed effect of filter clear poly bag cover in direct
inoculation screening of F. moluccana by fresh U. tepperianum spores as inoculums. Three months old of F. moluccana seedlings were used as host target. This research was designed as
four groups of cases, seedlings with cover and non-cover, gall rust from 400 m asl and from 800 m asl altitudes sites. Each group comprises three families with 10 individual seedlings as
replication. There were significant differences in percentage of galled formation at all treatments among family 1 and total families, but no significant effect at family 2 and 3 on
NC400 and NC800 treatments. The highest seedlings percentage of gall rust formation 94.2480 ± 1.97468 was in Family 2 with source of inoculums from 400 m asl. U.
tepperianum spores from lower altitude site source can more able to formed gall rust than the higher altitude site.
Keywords: F. moluccana, U. tepperianum, gall rust, filter paper, direct inoculation, screening
1. INTRODUCTION
Use of disease tolerantresistant plants is the ideal method to manage plant diseases, if plants of satisfactory quality and adapted to the growing region with adequate levels of durable
tolerance are available. The use of disease tolerance plants eliminates the need for additional efforts to reduce disease losses unless other diseases are additionally present. Tolerant plants
are usually derived by standard breeding procedures of screening Fry, 1982; Arneson, 2001; Maloy, 2005.
On a global scale, some of the most serious fungal plant pathogens are obligate biotrophic parasites Brown Hovmøller 2002. Obligate biotrophic U. tepperianum - F. moluccana
interactions were the main obstacle of gall rust disease tolerant screening. The term obligate biotroph characterizes a specific lifestyle in which the host suffers only minor damage. The
pathogen in turn is dependent on the living host plant to complete its life cycle Staples 2000. U. tepperianum produces teliospores which have ridged longitudinal striations, with three spores
on each head. The size of the teliospores ranged from 13-
18 m wide and 17-26 m long. The teliospores cannot themselves infect the host; they have to germinate to produce
basidiospores, which are formed at least 10 hours after inoculation. Then a penetration peg was formed by a matured basidiospore 16 hours after inoculation, penetrates the host cells
directly through the epidermis. Seven days after inoculation, vegetative mycelia of this gall-
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forming rust give rise to pycnia, recognized as small brown pustule which breaks through the epidermis. The typical symptom of gall rust disease on the seedlings is the bending of the stem
or shoot, either with or without the formation of a dark red necrotic lesion Rahayu, 2007. Symptoms of the disease begins with local swelling tumefaksi in the affected part of the plant
leaves, branches and stems, further swelling turned into lumps which then became a small pimple or called gall. Arising galls have varied from round to form irregular diameters ranging
from a few millimeters to greater than 10 cm. The galls can be grouped or spread on the affected areas. Young gall green light brownish covered by a layer of powdery slightly reddish
color which is a collection of spores. Old gall reddish brown to black, usually the gall is porous or perforated, and is used as a nest of ants and other insects. When the diseased
section of the petiole compound or canopy, that part a little bent because of thickening and swelling, then roll up the canopy leaves change shape malformations no longer leaves. If the
plants have severe attacks, then the whole plant is filled by the gall, then the leaves dry up experiencing hair loss, followed by the trunk and branches of trees and plants eventually die
Anggraeni, 2008. The germplasm screening methods can be classified into i direct intact or live plants, ii
direct detached plant organs, and iii indirect approaches Steadman et al., 1997; Olivier et al., 2008. Excised common inoculation of F. moluccana has been performed on direct inoculation
using fresh gall rust spores suspensions Morris, 1987; Kull et al., 2003. Effective screening for disease resistance requires accurate simulation of natural environmental conditions where
plants are exposed to the inoculums Porta-Puglia Aragona, 1997. Optimum inoculation and incubation conditions should be established so that susceptible and resistant genotypes
can be easily differentiated Infantino et al., 2006. Limited availability of space is often the major constraint to screening in environment control.
Disease evaluation in controlled conditions is often used to identify resistant breeding material during non-crop periods, but may also be used to confirm the reaction of tolerantresistant
genotypes identified in the field or for characterization of pathogen variability Infantino et al., 2006. Screening in the greenhouse or nursery without plastic cover allows challenging
inoculums in interaction with other phytopathogenic organisms. Even though a major problem using plastic covers are high moisture, low respiration and the opportunity for cross-
contamination via the wet surface Narciso, 2008. Filter papers were used for numerous laboratory applications filtration and exhibit such large
proportions of bacteria and spores in air flow or liquid. Filter paper was first used as a scientific tool in 1815 by the Swedish chemist Jöns Berzelius. Over the last 50 years, filter
paper has gained an increasingly important role as a substrate for the diagnosis and surveillance of infectious diseases Smit et al., 2014. Efficiencies for air filtration are normally
expressed as percent penetration or retention for a stated airborne particle size. Particle size for bacteria is between 0.22 µm
– 30 µm and 3.3 µm for spores General Electric Company, 2013. Filter paper on plant screening plastic cover can establish natural environmental
conditions for disease target development like humidity, respiration and temperature, without interaction with other phytopathogenic organisms.
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2. EXPERIMENTAL METHOD 2.1 Seedlings and gall rust spores material
