The conservation of endemic and endangered tree species Kalapia celebica Kosterm with Mycorrhizal bio-fertilizer technology in post-gold mining areas

Abstract

This research aimed to determine the effectiveness of mycorrhizal fungi of arbuscular (AMF) on the growth of Kalappia celebica Kosterm at 4 months aged after planting in gold mining land of PT. Panca LogamMakmur, Bombana, Southeast Sulawesi. This study was designed using block randomized design (BRD) consisted of 5 treatments, i.e. control, Constrictum etunicatum, Septoglomus constrictum, Glomus claroideum Schenk & Smith, Glomus coronatum Giovann, and mixed AMF (G. claroideum, G. coronatum) . Each treatment had three replications. The results showed that AMF colonization significantly increased the growth of K. celebica plants. G. coronatum and C. etunicatum significantly increased the height, diameter, and leaf area and leaves dry weight of K. celebica . AMF can be used for the conservation of extinct tree species in gold mining lands.

Full text

IOP Conference Series: Earth and Environmental Science
PAPER • OPEN ACCESS
The conservation of endemic and endangered tree species
Kalapia
celebica
Kosterm with Mycorrhizal bio-fertilizer technology in post-gold
mining areas
To cite this article: A Arif et al 2021 IOP Conf. Ser.: Earth Environ. Sci. 800 012039
View the article online for updates and enhancements.
This content was downloaded from IP address 216.19.200.184 on 12/07/2021 at 14:01

Content from this work may be used under the terms of the Creative Commons Attribution 3.0 licence. Any further distribution
of this work must maintain attribution to the author(s) and the title of the work, journal citation and DOI.
Published under licence by IOP Publishing Ltd
International Conference on Sustainable Utilization of Natural Resources 2020
IOP Conf. Series: Earth and Environmental Science 800 (2021) 012039
IOP Publishing
doi:10.1088/1755-1315/800/1/012039
1
The conservation of endemic and endangered tree species
Kalapia celebica Kosterm with Mycorrhizal bio-f
ertilizer
technology in post-gold mining areas
A Arif*, Husna*, F D Tuheteru and A Adiansyah
Department of Forestry, Faculty of Forestry and Environmental Science, Halu Oleo
University, Kendari, Southeast Sulawesi 93121, Indonesia
*E-mail: antyarf75@gmail.com
Abstract. This research aimed to determine the effectiveness of mycorrhizal fungi
of arbuscular
(AMF) on the growth of Kalappia celebica Kosterm at 4 months aged after planting in gold
mining
land of PT. Panca LogamMakmur, Bombana, Southeast Sulawesi. This study was designed using
block randomized design (BRD)
consisted of 5 treatments, i.e. control, Constrictum etunicatum,
Septoglomus constrictum, Glomus claroideum Schenk & Smith, Glomus coronatum Giovann, and
mixed AMF (G. claroideum, G. coronatum). Each treatment had three replications. The results
showed
that AMF colonization significantly increased the growth of K. celebica plants. G.
coronatum and C. etunicatum significantly
increased
the height, diameter, and leaf area and leaves
dry weight of K. celebica. AMF can be used for
the conservation of extinct tree species in gold
mining lands.
1. Introduction
Kalapi (Kalappia celebica Kosterm) is
a local plant that is currently being developed. This plant belongs
to the Fabaceae family, which has relatively
good
wood quality. Where the strength of wood included in
class II and durability also included in class III-II.
This timber is used as raw materials for house
constructions such as frames, doors, windows, and furniture with high economical prices. Because of
forest land diminishing as a results of converted to the mining, agricultural land, and
plantation, the
population of kalapi have also decreased.
Moreover, this plant was also exploited by people [1
,
2 , 3], so that their existence only remains in high
land in the forest just because of
difficult to be accessed. The impact, K. celebica plant was listed in the
IUCN red book and was
categorized
as an endangered species. Beside of external conditions, the decline
in K. celebica population was also caused by
internal
factors. The flowering and fruiting period occurred
very rarely only once in every three years, making it difficult
to obtain the seeds. The low quality of seeds
is also the reason for their poor germination. The good
thing
is K. celebica can be propagated asexually
through cuttings [4]. Based on research and experience, the cuttings
will grow easily if the material source
was taken from the juvenile tissues of seedlings than from the
mother
trees. Therefore, to maintain this
plant's existence, conservation efforts should be done with expansion of the
planting area such as in post
mining land.
Decreasing of forest area induce people to find other areas for planting locations such as post-mining
areas. However, the post-mining land has many obstacles. The land condition is an open area so that the
temperature at the location is very extreme with high temperature, drought, dense and rocky, as well as
low of macronutrients. In addition, high heavy metal content that included is also
makes
plants difficult to
grow well. The efforts which required to overcome this problem is the introduction of
biological
agents

International Conference on Sustainable Utilization of Natural Resources 2020
IOP Conf. Series: Earth and Environmental Science 800 (2021) 012039
IOP Publishing
doi:10.1088/1755-1315/800/1/012039
2
such as arbuscular mycorrhizal fungi (AMF). AMF will help plants grow well in marginal soil, such
as
nutrient-poor, drought, and high heavy metal content. [5] showed that AMF was effective and colonized
many
plants
on dry and degraded lands. Many researchs revealed that AMF could increase plant growth in
extreme, dry
and marginal lands by helping plants to obtain nutrients and water [2, 6, 7, 8], protecting
plants from soil pathogen attack and increasing tolerance of plants to heavy metals [7, 8] as well as
improving
physic
al and soil properties [9].
2. Materials and Methods
2.1. Field work
The research was conducted at gold
postmining land, PT. Panca Logam Makmur, Bombana, Southeast
Sulawesi in April-August 2020. After that, the observation was continued
in the Laboratory of Indonesia
Mycorrhiza Association (AMI) branched Southeast Sulawesi and Laboratory of Forestry, Faculty of
Forestry and Environmental Science, Halu Oleo University.
2.2. AMF inoculums and seedlings preparation
AMF inoculums were isolated from the
rhizosphere of Pericopsis mooniana [10]. These inoculums were
cultured on zeolite
media
with Pueraria javanica as host plants for three months and after that plants
were dried to trigger spores formation. K. celebica seeds were collected from mother trees in Abuki,
Konawe Regency, Southeast Sulawesi. Seeds were treated by soaking in hot water at 50oC temperature as
pretreatment
and then germinated on a plastic germinator pot. The polybag (size 15 × 20 cm) were filled
using mixed
media between sterile soil and tailing for growth media. The next, polybag media was holed
and put
plant
with AMF inoculums as much as 10 g around roots plants. The plant was watered every day
and
maintained
for seven months. Seedlings were grown in greenhouse of Indonesia Mycorrhiza
Asociation (IMA) branched Southeast Sulawesi.
2.3.
Transplanting
to the field
Seven months-aged Seedlings were transferred to the field with spacing 40cm × 40cm × 40 cm. The
experimental design used was Block Randomized Design (BRD) consisted of 5 treatments, i.e. control,
Constrictum
etunicatum, Septoglomus constrictum, Glomus claroideum Schenk & Smith, Glomus
coronatum Giovann, and mixed AMF (G.
claroideum, G. coronatum). Each treatment had three
replications.
2.4. Growth parameters
Observation were conducted for some
growth
variables including seedlings height (cm) was measured
using a ruler, starting from the stem's base to the stem
line's highest growing point. Seedlings diameter
(mm) was measured by using callipers as high as 1 cm above the media. The number of leaves (blade) was
measured by counting the number of leaves. Plants dry weight
was
determined at the end of research when
seedlings aged four months and were harvested for ten leaves each treatment (randomly selected) to keep
the plants intact to the field [11]. The leaves were oven to 70°C
for 2x24 hours or until constant weight
reached then dried leaves samples were weighed.
2.5. Mycorrhizal colonization
The
observation of the root colonization used the [12] modified method. Roots were washed and soaked in
10%
KOH
solution for 1 hour at 90oC. Then roots were washed with running water and soaked in 2% HCl
solution for 30 minutes. After that, the root samples were soaked in staining solution (0.05% trypan blue +
70%
glycerol + 30% distilled water) for 24 hours. Roots about 1 cm were arranged on preparatory glasses

International Conference on Sustainable Utilization of Natural Resources 2020
IOP Conf. Series: Earth and Environmental Science 800 (2021) 012039
IOP Publishing
doi:10.1088/1755-1315/800/1/012039
3
and observed under microscope. Percentage of AMF colonization was determined with the formula
[Σmycorrhizal viewpoint /
Σtotal viewpoint observed] × 100% [5].
2.6. Statistical analysis
The results data were analyzed using variance (ANOVA) with the F test. if the test has a significant effect,
then continued by the Duncan test
at the 95% confidence level. Data analysis was conducted using SAS
software.
3. Results
3.1. Mycorrhizal colonization
AM fungi colonized all of the seedlings roots of K.celebica (Figure 1). Histogram showed that in general,
AMF treatments gave higher percentages of colonization than control, and the highest percentage on
mixed AMF treatment.
Figure 1. Percentage of AMF colonization in roots of 4 months K. celebica plants in gold mining land
3.2. Plant growth
In general, AMF treatments resulted in better values trend in all of growth
variables
than control.
Inoculation roots plant with G. coronatum gave the highest value and increased plant height, stem
diameter of K. celebica. Furthermore, G. coronatum and mixed AMF treatments also increased the
number of leaves (table 1).

International Conference on Sustainable Utilization of Natural Resources 2020
IOP Conf. Series: Earth and Environmental Science 800 (2021) 012039
IOP Publishing
doi:10.1088/1755-1315/800/1/012039
4
Table 1. Averages of plant height, stem diameter and number of leaves on the inoculated and
uninoculated
plant
of K. celebica aged four months on gold mining land
Treatments
plants height (cm)
Stem Diameter (mm)
Number of leaves (blades)
Control
9.5 b
1.74 c
34.3 b
C. etunicatum
15.2 b
2.51 bc
54.0 b
S. contrictum
11.4 b
2.57 bc
39.0 b
G. coronatum
24.6 a
4.70 a
102.3 a
Mixed AMF
15.6 b
3.72 ab
104.0 a
Notes: Average values followed by different letters in
the same column are significantly different at Duncan test
(P<0.05)
Variables of length and width leaf, leaves
dry weight of K. celebica colonized by C. etunicatum gave
the highest values than other treatments. While control treatment gave the least value (table 2).
Table 2. Averages of leaf length, leaf width, and leaves
dry weight on K. celebica aged four months on
gold mining land
Treatments
Leaf length (cm)
Leaf width (cm)
Leaves dry weight (g)
Control
4.14 b
1.88 c
0.55 c
C. etunicatum
14.49 a
6.14 a
3.77 a
S. contrictum
9.48 a
4.33 ab
2.00 b
G. coronatum
10.63 a
5.20 ab
2.86 ab
Mixed AMF
10.54 a
4.15 b
2.25 b
Notes: Average values followed by different
letters
in the same column are significantly different at Duncan test
(P<0.05)
4. Discussion
Plant growth on gold postmining land
experienced many obstacles due to extreme land conditions. The
characteristics of land and unfavorable climate conditions are the challenging
that should be solved. The
soils are dense and dry in the summer and inundated in the rainy season. In
addition,
unfertile land and
high heavy metals content also the important problems that make delaying of plants growth. High
temperatures
was recorded when study conducting on the field ranging from 35-40oC and even up to 40oC.
This condition made plants difficult to grow and develop well. To solve this, some of additional
treatments were required to improve soil fertility and heavy metals plant tolerant, and the using of paranet
was also required to reduce the light
intencity
and temperature.
The selection of suitable plants species and benefit soil microorganism are very critical. The use of
biological agents such as AMF, which is inoculated to plants, can assist plants to adapt and to grow
optimally
on land. It was proved by many research that mycorrhizal plants can adapt to extreme
environmental conditions such as dried
condition, infertile land, saline soil condition etc. [5]. That is why,
AMF selection on plant types and soil under certain conditions was required to get the best suitable

International Conference on Sustainable Utilization of Natural Resources 2020
IOP Conf. Series: Earth and Environmental Science 800 (2021) 012039
IOP Publishing
doi:10.1088/1755-1315/800/1/012039
5
inoculums to increase plants growth. The results revealed that, in general, AMF gave the better growth of
plant, so its application is important to support the development of kalapi plants. This is the first study of
AMF colonization in K. celebica in gold mining land to the best of our knowledge. Plants that colonized
by AMF had better growth compared to uninoculated plants. This can be seen in some of growth variables
consisted
of plant height, diameter, leaves number, length and width leaf, as well as leaves dry weight.
Similar with [8, 7, 13], AMF inoculated seedlings showed better growth than non inoculated seedlings.
From several types of AMF treatment, data showed that the best AMF treatments were G. coronatum and
C. etunicatum and followed by mixed AMF, which significantly increased the height, diameter, and
number of K. celebica leaves. This research was
also
in line with the research conducted by [14] that AMF
is very important and can give
positively
affect to Ploiarium alternifolium and Calophyllum hosei plants
in their initial growth. AMF helps plant growth by increasing
the absorption of nutrients, especially
phosphorus and water, and increasing plant tolerance to heavy metals [6, 15
]
. The increase in leaf area is
due to the sufficient nutrients obtained from plants association with AMF. Moreover
,
the photosynthesis
process in mycorrizal plants runs well, which will increase the accumulation of total dry weight in
mycorrhizal plants [16, 17, 18]. The plants which was colonized by AMF marked by fungi structure such
as internal and external hypha, mycelium and vesicles in roots plants. Determining the percentage of
colonization in roots plant was carried out by the staining method [12]. It was seen that the highest AMF
colonization was given by mixed AMF treatment and then followed by other AMF treatments. The lowest
percentage of
colonization
was showed in the control treatment. Plants which grow better will give
positive affect on growth of AMF and other microbes population Fotosynthates products produced from
photosynthetic process will be used for tissues formation and
exuded
as roots exudates. It is used as
energy sources for AMF and other microbes. According to [19
]
, plant roots exude chemical compounds
such as amino acid, carbohydtrate, organic acid, hormone and etc. which will
stimulat
e growth of
beneficial microbes on roots. Based on data, uncertain field conditions resulted the colonization about
30%.
Meanwhile, research in the greenhouse conducted by [8] using gold mining tailings were obtained
root colonization of
K.
celebica plant was about 70%. The suitability between AMF and host plants is
influenced by the host plant
itself,
type of AMF and environmental conditions.
5. Conclusion
In conclusion, AMF colonization significantly increased the growth of
K.
celebica plants. Types of G.
coronatum and C. etunicatum significantly increased the height, diameter, and leaf area and
leaves dry
weight of K. celebica. Therefore, AMF can be used for conservation of endangered tree species in post
-
gold mining land.
Acknowledgement
The authors are thanks to the Ministry of Research and Technology, Higher Education of
the
Republic of
Indonesia (RISTEK DIKTI) for funding support. The authors are also thanks to the Chairman of PT.
Panca
Logam Makmur for providing research site.
References
[1] Arif A, Tuheteru F D, Husna, Kandari A M, Mekuo I S and Masnun 2016 Status and culture of
arbuscular mycorrhiza fungi isolated from rhizosphere of endemic
and
endangered species of
kalapi (Kalappia celebica Kosterm). European Journal of Sustainable Development 2(2): 395-
402.

International Conference on Sustainable Utilization of Natural Resources 2020
IOP Conf. Series: Earth and Environmental Science 800 (2021) 012039
IOP Publishing
doi:10.1088/1755-1315/800/1/012039
6
[2] Husna, Mansur I, Budi S W R, Tuheteru F D, Arif A, Tuheteru E J, and
Al
Basri. 2019 Effects of
arbuscular mycorrhiza fungi and organic material on growth and nutrient uptake by Pericopsis
mooniana in coal mine. Asian Journal of Plant Sciences 18(3):101-9.
[3] Liam A T, Arif A, Clark R P, Girmansyah D, Kintamani E, Prychid C J, Pujirahayu N,
Rosmarlinansiah, Brearly F Q, Utteridge T M A, and Lewis G P 2019 An enigmatic genus on an
enigmatic island: the re discovery of Kalappia on Sulawesi. Ecology 100 (11)
[4] Arif A, Tuheteru, F D, and Husna 2015 The conservation of endemic and endangered species Kalappia
celebica Kosterm through cutting propagation and arbuscular mycorrhiza fungi (AMF) potential
assessment. Int.
Conf
. Sustainable Development Proc.. p 53-65.
[5] Bhale U N 2018 Arbuscular mycorrhizal fungi (AMF) s
tatus
and diversity of weedy plants in degraded
land. International Journal of Plant Pathology 9(1):1-8
[6] Smith
S E, and Read D J 2008 Mycorrhizal Symbiosis. 3rd Edition (London: Academic Press) p 800
[
7
] Husna, Tuheteru F D, Arif A 2017 Arbuscular m
yc
orrhizal fungi and plant growth on serpentine
soils. In: WuQS (Eds.), Arbsucular mycorrhiza
s
and stress tolerance of plants. (Singapore:
S
pringer) p 293-303
[8] Husna, Tuheteru F D, and Solomon 2019 Improvement of early growth of endemic Sulawesi tree
species K. celebica by arbuscular mycorrhizal fungi in gold mining tailings. IOP Conf.
Series:
Earth and Environmental Science, IOP Publishing, 394
[9] Agus C, Primananda E, Faridah E, Wulandari D,
and
Lestari 2018 Role of arbuscular mycorrhizal
fungi and Pongamia pinnata for revegetation of tropical open-pit coal
mining
soils.
International Journal of Environment Science and Technology. 16: 3365-74
[10] Husna, Budi
S
W R, Mansur I, and Kusmana C 2015 Respon pertumb
uh
an bibit kayu kuku
(Pericopsis mooniana (Thw.) Thw.) terhadap inokulasi fungi mikoriza a
r
buskula lokal.
Pemuliaan Tanaman Hutan 9(3):
13
1-48
[11] Giri B, Kapoor R, and Mukerji K G 2005 Effect of the arbuscular mycorrhizae Glomus
fasciculatum
and G. macrocarpum on the growth and nutrient content of Cassia siamea in a semi-arid Indian
waste land soil. New Forest. 29:63–73.
[12] Brundrett M, Bougher N, Dell B, Grove T, and
Malajczuk
N 1996 Working with Mycorrhiza in
Forestry and Agriculture. (Canberra, Australia: Australian Centre for International Agricultural
Research) p 374
[13] Husna, Tuheteru F D, and Arif A 2018
Arbuscular mycorrhizal fungi symbiosis and conse
r
vation of
endangered tropical legume trees. ed
Giri et al., Root Biology, Soil Biology (Ger
m
any:
Springer) p 52
[14] Turjaman M, Tamai Y, Sitepu I R, Santoso
E, Osaki M, and Tawaraya K 2008 Improvement of early
growth of two tropical peat-swamp forest tree species Ploiarium alternifolium and Calophyllum
hosei by two arbuscular mycorrhizal fungi under green house conditions. New Forest 36: 1-12
[15] Yang Y, Han X, Liang Y, Ghost A, Chen J, and Tang M 2015 The combined
effects
of arbuscular
mycorrhiza fungi (AMF) and lead (Pb) stress on Pb accumulation, plant growth parameters,
photosynthesis, and antioxidant enzymes in Robinia pseudoacacia L. Plos One, 10(12)
[16] Wulandari D, Saridi, Cheng W, and Tawaraya K 2014 Arbuscular mycorrhiza colonization enhanced
early growth of Mallotus paniculatus and Albizia saman under nursery conditions in E
ast
Kalimantan, Indonesia. International Journal of Forestry Research 1-8
[17] Husna, Tuheteru
F D, and Wigati E 2017 Growth response and dependenc
y
of endangered nedum tree
species (Pericopsis
m
ooniana) affected by indigenous arbuscular m
yc
orrhizal fungi inoculation.
Nusantara Biosci
e
nce 9(1) : 57-61
[18] Husna, Tuheteru F D, A
r
if A, and Sera G R 2018 Respon pertumbuhan dan ketergantungan jenis
legume terancam punah angsana (Pterocarpus indicus Willd) terhadap aplikasi fungi mikoriza

International Conference on Sustainable Utilization of Natural Resources 2020
IOP Conf. Series: Earth and Environmental Science 800 (2021) 012039
IOP Publishing
doi:10.1088/1755-1315/800/1/012039
7
arbuskula lokal. Prosiding Seminar Nasional Silvikultur V dan Kongres Masyarakat Silv
i
kultur
IV. Lambung Mangkurat University Pre
s
s. Banjarmasin.
[19] Osman K T 2013 Forest Soils, Properties and mMnagement. (Switzerland: Springer
International
Publishing) p 217