Научная статья на тему 'Growth response of Tabernaemontana pandacaqui Poir. in soil from a former mining site in Marinduque, Philippines'

Growth response of Tabernaemontana pandacaqui Poir. in soil from a former mining site in Marinduque, Philippines Текст научной статьи по специальности «Биологические науки»

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Ukrainian Journal of Ecology
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mining site / mycorrhiza / Acacia auriculiformis / Tabernaemontana pandacaqui / growth response

Аннотация научной статьи по биологическим наукам, автор научной работы — A.J.D. Belarmino, N.M. Pampolina, E.E. Coracero

Potential hazards and immediate rehabilitation of inactive/abandoned mines are concerns in Mogpog, Marinduque. Efforts have been initiated in the area through the use of species such as Acacia auriculiformis Benth. The study focused on assessing and mass-producing endomycorrhizal fungi from plants collected in the area and investigating the growth response of indigenous Tabernaemontana pandacaqui Poir. with A. auriculiformis, endomycorrhizas, and biofertilizers in soil from mine tailings. Soil and plant collection was done in three major ecosystems: agroforestry, A. auriculiformis stand, and natural forest. Associated endomycorrhizas were isolated and prepared as an inoculant to test the response of T. pandacaqui. Field endomycorrhizal spores from T. pandacaqui were: Acauslospora (50), Glomus (16), and Gigaspora (2), which sporulated (422/75g dry soil) as inoculant. Height and diameter of T. pandacaqui were significantly (p < 0.01) better with A. auriculiformis, possibly due to its N-fixing property, though host mortality was higher brought by competition. Soil inoculants promoted the growth of T. pandacaqui but MykoVAM performed significantly (p < 0.05) in height than control. To facilitate the ecological succession in old mining, the A. auriculiformis could be maintained as a nurse crop for threatened species like T. pandacaqui and consider the potential of local endomycorrhizal isolates encouraging growth and survival.

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Текст научной работы на тему «Growth response of Tabernaemontana pandacaqui Poir. in soil from a former mining site in Marinduque, Philippines»

Ukrainian Journal of Ecology

Ukrainian Journal ofEcology, 2021, 11(2), 111-121, doi: 10.15421/2021_87

ORIGINAL ARTICLE

Growth response of Tabernaemontana pandacaqui Poir. in soil from a former mining site in Marinduque, Philippines

AJ.D. Belarmino1 , N.M. Pampolina1 , E.E. Coracero1'2*

1 College of Forestry and Natural Resources, University of the Philippines Los Baños, Laguna, Philippines 2Forestry and Environmental Sciences Department, Aurora State College of Technology,

Baler, Aurora, Philippines Corresponding author E-mail: ericsoncoracero@ascot. edu.ph

Received: 14.03.2021. Accepted: 14.04.2021.

Potential hazards and immediate rehabilitation of inactive/abandoned mines are concerns in Mogpog, Marinduque. Efforts have been initiated in the area through the use of species such as Acacia auricuifformis Benth. The study focused on assessing and mass-producing endomycorrhizal fungi from plants collected in the area and investigating the growth response of indigenous Tabernaemontana pandacaqui Poir. with A. auricuifformis, endomycorrhizas, and biofertilizers in soil from mine tailings. Soil and plant collection was done in three major ecosystems: agroforestry, A. auricuifformis stand, and natural forest. Associated endomycorrhizas were isolated and prepared as an inoculant to test the response of T pandacaqui. Field endomycorrhizal spores from T pandacaqui were: Acausiospora (50), Glomus (16), and Gigaspora (2), which sporulated (422/75g dry soil) as inoculant. Height and diameter of T pandacaqui were significantly (p < 0.01) better with A. auricuifformis, possibly due to its N-fixing property, though host mortality was higher brought by competition. Soil inoculants promoted the growth of T pandacaqui but MykoVAM performed significantly (p < 0.05) in height than control. To facilitate the ecological succession in old mining, the A. auricuiiformis could be maintained as a nurse crop for threatened species like T pandacaqui and consider the potential of local endomycorrhizal isolates encouraging growth and survival.

Keywords: mining site, mycorrhiza, Acacia auricuiiformis, Tabernaemontana pandacaqui, growth response.

Introduction

Mineral resources are one of the Philippines' acknowledged rich assets that significantly contribute to the country's economic development.Marinduque is among the mineral-rich provinces with large copper deposits discovered by mining prospectors in the late 1960s. It became the home to one of the most significant mining operations in the country. However, a disastrous mine tailing accident in 1996 affected the provincial island (Gregory, 2004)). Republic Act 7942 or the Philippine Mining Act of 1995 states that "Contractors and permittees shall technically and biologically rehabilitate the excavated, mined-out tailings covered, and disturbed areas to the condition of environmental safety, as may be provided in the implementing rules and regulations of this Act". In this regard, the Department of Environment and Natural Resources (DENR) requires responsible rehabilitation of mined areas (Teves, 2018). At least 22 inactive and abandoned mine sites need a plan for rehabilitation like the case of an inactive copper mining site in Mogpog, Marinduque. The government and mining companies are making efforts to restore mined-out areas and convert areas as productive assets (Felix, 2004). The mountain situated in the area was flattened and replaced with a mountain of dogged unprocessed copper-rich soil that reached adjacent rice fields, agricultural areas, and mangrove forest. The mining activities led to several environmental problems, such as deforestation, leaving only less than 30% of the forest cover in 2001 (Medianista & Labay, 2017). Such events caught the Philippine government's attention and made them prioritize the rehabilitation and remediation of the inactive mine sites to enhance its current state. Thus, improving biodiversity can help in natural ecological processes and human consumption (Coracero & Malabrigo, 2020).

Phytoremediation is an emerging biological approach in the cleanup of contaminated sites. According to Sigurdur Greipsson (2011), phytoremediation is a recently developed technology that offers a cost-effective solution by using plants, and associated soil microbes, to reduce the content, or toxic effects, of contaminants in the environment. It is a remediation technique that takes advantage of the biological processes of plants. Plants alter the soil through theshoot and root growth, water and mineral acquisition, senescence, and eventual decay. The following series of actions directly or indirectly absorb, sequester, and/or degrade contaminants. Planting and growing compatible species either remove the pollutant or physically or chemically convert contaminants into a component that no longer poses a threat to the surrounding environment (Cunningham & Ow, 1996). In the Philippines, DENR uses phytoremediation species such as Acacia auricuifformis Benth., Acacia mangium Willd., Pterocarpus indicus Willd., Casuarina equisitffoia Forst., and Chrysopogon zzzanioides (L.) Roberty. The Philippine governmentPhilippine government supported the department's initiative supported the department's initiative, thus issuing Executive Order 270 entitled "National Policy Agenda in Revitalizing Mining in the Philippines" on January 16, 2001 and stated in Section 2-I, that "Remediation and Rehabilitation of abandoned mines shall be accorded top priority to address the negative impacts of past mining projects." (DENR, 2014). One of major steps under the executive order was the formation of the Bioremediation Research Team through National Academy for Science and Technology to conduct studies and research to develop technologies that can help in the remediation of toxic mining wastes (Raymundo et al., 2006).

The DENR initiated to plant A. auricuiiforms, also known as earleaf acacia, in the area in 2009. According to the ERDB Director Dr. Henry A. Adornado, 42 ha of mined-out areas were successfully planted. It was found that A. auricuiiforms and other species that

were planted for the phytoremediation, such as Pterocarpus indicus Willd. were most efficient in the absorption of metals (Sarian, 2017). There were no records on the number of individuals of these species the DENR have planted throughout the years. Nevertheless, patches of A. auricuiiformis can be observed and have dominated the area. Earleaf Acacia is a fast-growing introduced species that can grow on poor quality soil. Earleaf acacia has dense roots and tolerates seasonally waterlogged soil and produces a heavy leaf litter, making it suitable for rehabilitation of degraded land (Contu, 2012). These characteristics make the species compatible for stabilizing eroded land. It is popular in stabilizing and revegetating mined-out areas because A. auricuifformis can grow in infertile soil and tolerate both highly acidic and alkaline soils capable of nitrogen fixing (Orwa et al., 2009). This study aims to assess the effects of A. auricuiifbrmis to the growth of Philippine native species in Mogpog, Marinduque, such as Tabernaemontana pandacaqui (Pandakaki). Specifically, the study aims to assess and mass produce endomycorrhizal fungi of plants in the area and determine the growth behavior and response of Pandakaki with Earleaf Acacia, endomycorrhizas, and soil amendments (inclusion of biofertilizers).

Methodology

Study site

Inventory and samples were gathered in Brgy. Capayang, Mogpog, Mariduque and is located at 13°30'16"N 121°51'42"E (Fig. 1a). Post mine areas have soils contaminated with acid mine drainage (Medianista & Labay, 2017).

The province was known for its rich mineral deposits such as gold, copper, and iron. During the 1960s up to the late 1990s, mining activities were operated on-site in open-pit mining. All mining activities on the island were ceased when the 1.5-3 million cubic meters of sulfidic tailings slurry from the Tapian Pit storage area into the Makulapnit River, Boac River, and eventually the ocean west of the island (Plumlee et al., 2000). The mining caused several environmental issues brought about by improper mining activities, such as mine tailings being dumped in rice fields, mangroves, agricultural lands, and forest lands. The mine site in Brgy. Capayang, Mogpog, Mariduque is a copper-rich soil. Soil analysis from the study of Tulod et al. (2012) showed a high concentration of copper and low content of other toxic contaminants (Table 1). The soil in the area was a loam type soil with 62% sand, 36% silt, and 2% clay and highly acidic (pH = 4.4) with deficient nitrogen (100 ppm), potassium (101.4 ppm), phosphorus (81 ppm), OM content (0.24%), and CEC (17.7 cmol/kg).

Table 1. Heavy metals present in the soil of inactive/abandoned mining sites (Tulod et al., 2012).

The growth of vegetation in this kind of condition is nearly unattainable. Excess levels in minerals in soil lead to abnormality in plant growth. The ideal pH for plant growth ranges from 5.5 to 7.0, although plants have adapted to withstand pH beyond this range. It is significant in many chemical processes in the soil, such as plant nutrient availability (The Mosaic Company, 2020). To address the condition in the inactive/abandoned mining site, the DENR initiated the site's rehabilitation, and one of their actions was planting A. auriculiformis. The area is now covered with clumps of A. auriciformis, and other plant species are starting to thrive in the copper-rich soil (Fig. 1b).

Selection of species for the experiment

The vegetation composition of the area was considered in selecting the appropriate species to test. Thus, vegetation assessment was conducted in the area. Three types of ecosystems were surveyed: the natural forest, agroforestry ecosystem, and the Earleaf Acacia stand. Acacia auricuifformis A. Cunn. ex Benth. and Tabernaembntana pandacaqui Poir. were found to be the most populous for trees and understorey species, respectively. A. auriculiformis is an introduced species used in the rehabilitation and phytoremediation, while T. pandacaqui is a Philippine native species thriving in the area. The effect of their coexistence in an ecosystem is an interesting aspect to study. Therefore, these two species were used as the subjects of the experiment.

Isolation, characterization, and mass production of endomycorrhizas

Eight (8) abundant wildling species were collected still attached to its soil at the rhizosphere layer. These species were Trema orientals (L.) Blume, Guba kbelreuteria (Blanco) Merr., Antidesma ghaesembiHa Gaertn., Morinda citrifoHa L., T. pandacaqui, Pittbspbrumpentandrum (Blanco) Merr., Alstonia macrbphylla Wall. ex G. Don, and A. auricuiffbrmis. For each sample, 25 grams of soil closest to the host plant's fine roots was subjected under the sieving and centrifugation method as stated in the book of Brunrdett et al. (1996). The soil is then continuously washed with water and sieved in 150 pm and 45pm mesh. After sieving, the remaining soils were placed in centrifuge tubes labeled with the corresponding mesh size, and then water was added. These tubes were placed in a centrifuge for 5 minutes at 2000 revolution per minute (RPM). Next, floating sediments were drawn off. Then, sucrose was poured into the centrifuge tubes. Once again, the tubes were placed in the centrifuge and were left to spin for 2 minutes at 2000 RPM. Following this step, floating sediments were sieved, and soils that settled at the bottom of the centrifuge tubes were discarded. Sieved soils were then placed in a petri dish. All were examined under the microscope and were isolated. The genus and individual count of the spores were recorded. The same procedures were done for each soil sample for the plant species collected. All isolated spores were then mass produced at UPLB BIOTECH laboratory and harvested with Centrbsema pubescens Benth. as host plants.

Metal Elements

Concentration (ppm)

Lead

Arsenic

Mercury

Zinc

Copper

Nickel

Chromium

Cadmium

4 13 3

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Fig. 1. Satellite image of mining area (a) (Google Earth, 2019) and the study site (b) in Brgy. Capayang, Mogpog, Marinduque.

Experimental set-up and observation

The interaction of T. pandacaqui and A. auricuiiformis under different soil amendments were observed from January 4, 2018, to April 4, 2018. To have an in-depth observation of the influence of A. aurccuifformis to T pandacaqui, some 50 T pandacaqui were planted individually in 4 x 8 cm polyethylene bags, and 50 T. pandacaqui with A. auricuiiformis were planted together in 4 x 8 cm polyethylene bags (Fig. 2). Sterilized soils collected from the old mining site were used as planting media. Both T pandacaqui with and without A. auricuiiformis were subjected to five soil amendments with ten replicates for each amendment. The following treatments were done, and all arranged based on CRD:

a. Control - sterilized soil only

b. Marinduque inoculants - sterilized soil plus the mass-produced endomycorrhizal inoculant

c. MykoVAM - sterilized soil plus MykoVAM biofertilizer (a biofertilizer produced by UPLB Biotechnology which aims to aid in nutrient and water absorption)

d. BIO-N - sterilized soil plus BIO-N biofertilizer (a biofertilizer produced by UPLB Biotechnology which has effective nitrogen supplement for plants)

e. The combination of all treatments - sterilized soil plus massed produced endomycorrhizal inoculant, MykoVAM, and BIO-N.

Fig. 2. Experimental layout showing T. pandacaqui (a) and set up with A. auriculfformis () *(Note for colored tags: Control (white), Marinduque inoculants (blue), MykoVAM (green), BBio-N (yellow), and combination (red))

Initial root, height, and diameter were recorded for all plants. The plant's height and diameter were measured in the level of the half of the height of polyethylene bag at least once a week. Changes in the diameter, height, and distinguishable physiological differences of the plants were recorded.

Results and discussion

Endomycorrhizal assessment and mass production

The endomycorrhizal fungi isolated from sampled ecosystem were characteristically microscopic that ranged from 45-150 pm. Their colors were transparent to white, pale to dark yellow, orange to dark orange, or brownish to dark brown. Some spores were doublewalled with shapes characteristically spherical, oblongate, ellipsoidal, or irregular. Other endomycorrhizal isolates exhibited hyphal attachments that were connected to fine roots (Table 2 & Fig. 3).

There were only three genera of endomycorrhizal fungi that were analysed in terms of spore density across all plots. Endomycorrhizal spores consisted of 228 Acaulospora (54%), 116 Acaulospora with hyphae (28%), 73 Glomus (17%), and 5 Gigaspora (1%) (Fig. 4). Acaulospora was the most prevalent among the AMF isolated.

Based on host plants, species A. auriculiformis had the highest spore count among the few soil samples of various plant species. A total of 101 individual counts of endomycorrhizal fungi were isolated from the rhizosphere of A. auricuifformiis (Fig. 5). The spores per 75 g of dry soil comprised of Acaulospora (71) and Glomus (30). The number of spores isolated from rhizosphere per 75 g of dry soils of T. pandacaqui comprised of Acaulospora (50), Glomus (16), and Gigaspora (2).

Table 2. Morphological description of endomycorrhizal genera isolated from different host plants and vegetative genera.

Host Plants Endomycorrhiza Genera

Acaulospora

T orientais

Gigaspora Glomus

G. koelreuteria Acaulospora A. ghaesembilla Acaulospora

Acaulospora

M. cttrffoiia

Gigaspora Acaulospra

T pandacaqui

Glomus Acaulospra

P. pentandrum

Glomus Acaulospra

A. macrophylla

Glomus Acaulospra

A. auriculfformis

Glomus

Morphological Description of Endo- mycorrhizal Spores

Spores collected were relatively small; have colors ranging in yellow, orange, and brown; double-walled; shapes observed were spherical and oblongate, and some have hyphal attachments. It was dark orange and spherical.

Collected Glomus was transparent to dark brown;

double-walled, and have spherical and irregular shapes.

Spores were yellow to orange in color; ellipsoidal, and

some of them has hyphal attachment.

These were yellow to dark yellow colored, transparent,

and some have hyphal attachment.

Some spores were pale yellow to dark yellow, and

others were dark orange; shapes observed were

ellipsoidal and spherical, and some have hyphal

attachments.

These were dark orange and spherical.

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Most were yellow, some are dark orange, and some were brownish to dark brown in color; double-walled; have an ellipsoidal and spherical shape, and some have hyphal attachment.

Spores were transparent to dark brown, double-walled, and spherical, and irregular in shape. These were pale yellow to dark yellow, and some were brownish; transparent; double-walled; spherical, and oblongate; and some have hyphal attachments. Color ranges from light brown to dark brown and relatively small.

Spores were in different shades of yellow, orange, and brown; double-walled; spherical, ellipsoidal, and oblongate in shape; and some have hyphal attachments. Observed spores were transparent to dark brown; double-walled; spherical, and irregular in shape. Transparent to dark yellow, orange, and brownish in color; double-walled; with shapes spherical, oblongate, and irregular; some have a hyphal attachment. Observed spores were transparent to dark brown; double-walled; spherical, and irregular in shape._

Fig. 3. Samples of (a) Acaulospora, (b) Gigaspora, (c) Glomus, and (d) hyphal attachment of endomycorrhizal isolates from rhizosphere in inactive/abandoned mining site in Brgy. Capayang, Mogpog, Marinduque.

1%

__ 17% ■ Acaulospora

28% 54% ■ Acaulospora with hyphae

■ Glomus

✓ ■ Gigaspora

Fig. 4. The proportion of endomycorrhizal genera of spores isolated from rhizosphere across vegetation formerly mined from Brgy. Capayang,

Mogpog, Marinduque.

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5. Total number of isolated spores (per 75 g dry soil) from different host plants growing within ecosystems in

inactive/abandoned mine

The diversity of endomycorrhizal fungi is significantly influenced by nutrient composition and soil texture varying in capacity and tolerance (Zhao, et al., 2017). Certain species are drought resistant, tolerant to pathogens, and could endure extreme soil temperature (Aggangan et al., 2015). Acaulospora, Glomus, and Gigaspora are particularly tolerant to acidic and/or alkaline soils (Clark, 1997). These species are resistant to harmful soil conditions like the mine site left in Brgy. Capayang, Mogpog, Marinduque. According to Lovelock et al. (2003), endomycorrhizal fungi are interdependent with plant roots. Growth rates or productivity could influence the sporulation of fungal symbionts. A greater volume of spores was observed with fast-growing species. A. auriculfformis is a fast-growing species, and its physiological characteristics affected the abundance of endomycorrhizal fungi. After mass production, the number of isolated spores gathered from the rhizosphere of 75 g dry soil of T pandacaqui was comprised of 212 spores in 150 pm sieve, and 228 in 45 pm sieve collected and were utilized as soil amendments for the experimental setup. Spores that were isolated were relatively large, have a range of colors from yellow to dark orange and light to dark brown, and the shapes were oblongate and ellipsoidal (Table 3), and the majority were Acaulospora species.

Table 3. Endomycorrhizal fungi (spore/ 25 g dry soil) isolated from rhizosphere of T pandacaqui with sizes and description.

Soil Sample

Endomycorrhizal genera 45 pm 150 pm

Description

Replicate 1

Acaulospora

Glomus

Replicate 2 Acaulospora Acaulospora

Replicate 3

Glomus

Total Mean

52

36

43 64

33

228 46

41

32

62 56

21

212

42

Double-walled, light yellow to dark brown, oblongate, transparent

Spores were transparent to dark brown, double-walled, and spherical, and irregular in shape

Mostly yellow, others transparent to white, ellipsoidal Yellow to dark brown, oblongate, double-walled Transparent to dark brown; double-walled; spherical and irregular in shape

There were no reports and/or studies found discussing the symbiotic relationship between T pandacaqui and endomycorrhizae. It is possible that the two have no special interaction, and it only exhibits only typical association of plants and endomycorrhizae. Nevertheless, the isolated spore may be accounted for T pandacaqui to in such an environment as the copper-rich soil of the former mining site in Mogpog, Marinduque. Aggangan and Cortes (2018) stated that endomycorrhizal fungi enhance plant tolerance to heavy metal contamination and induced Cu retention in the roots of the seedlings. Indigenous endomycorrhizal fungi have the potential in the rehabilitation of mined-out areas in the Philippines.

Growth Response of T. pandacaqui

Growth and survival of T. pandacaqui with A. auricuiiformis and amendments were not significant in height and diameter at first month. Development of T pandacaqui at 60 days after inoculation showed 50% mortality when planted with A. aurccuifformss. Growth of T pandacaqui was improved when applying MykoVAM than all other amendments. At harvest, the growth of T pandacaqui was better with A. auncuifformis. The height of T pandacaqui with and without A. auricuifformis was significantly different (p < 0.01). Height measurement of T pandacaqui with A. auricuiiformis in the control was the highest and grew by 21%. And the highest height value of T pandacaqui without A. auricuiiformis was MykoVAM by 11% among all treatments (Fig. 6). Height increment every 30 days of T pandacaqui with A. auricuiiformis applied with MykoVAM had the highest increment and by the end of the observation, it grew by 22% (Fig. 7a). Control and MykoVAM was significantly different (p < 0.05). The response of T pandacaqui without A. auricuiiformis to Bio-N yielded the highest growth increment in height by 14% after 30 days, and from the second month until harvest, control had the highest growth increment by 11% (Fig. 7b). Both Bio-N and Control have significant differences (p < 0.05).

8.00

Control

MI MycoVAM Bio-N Treatments

T.pandacaqui alone

T.pandacaqui with A. auricuiiformis

(a)

Fig. 6. Final Height of T pandacaqui in response to inoculation of amendments when planting with or without A. auricuiiformis (Note: MI- refers to Marinduque inoculants; All-means combination of all treatments)

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60 Days

90 Days

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Control Marinduque Inoculant MycoVAM Bio-N Combination of all treatments

0.00 0.64 1.40 0.70 1.27

0.00 0.64 9.27 1.59 1.27

0.00 0.76 3.43 1.91 1.02

Treatments

130 Days ■ 60 Days ■ 90 Days

(b)

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■ 30 Days 0.48 0.47 0.25 0.68 0.34

■ 60 Days 1.07 0.42 0.38 0.76 0.47

■ 90 Days 1.19 0.72 0.13 0.68 0.55

Treatments

■ 30 Days ■ 60 Days ■ 90 Days

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Fig. 7. Height Increment of T pandacaqui in response to inoculation of amendments when planting (a) with and (b) without A. Auriculiformis.

On the contrary, T. pandacaqui without A. auriculfformis had a superior final diameter measurement and was significantly different from T pandacaqui with A. auriculiformis (p < 0.05). Both Control and MykoVAM were the top treatments for the diameter (Fig. 8). In T pandacaqui with A. auriculfformis, MykoVAM had the highest increment for 2 months by 6%, and after 90 days, Marinduque inoculant had the highest diameter increment valuing 38% from the initial (Fig. 9a). The diameter of T pandacaqui alone, Bio-N has the highest diameter increment in 30 days by 14%, in 60 days, Marinduque inoculants had the highest increment valuing 34%, and combination of all treatments after 90 days with 60% increment from the initial planting; no significant difference was observed (Fig. 9b).

0.35

0.30

0.25

-Ü 0.20

£ 0.15

0.10

0.05

0.00

T.pandacaqui alone

T.pandacaqui with A. auriculiformis

Control MI MycoVAM Bio-N All

Treatments

Fig. 8. The final diameter of T pandacaqui in response to inoculation of amendments when planting with or without A. auriculiformis (Note: MI- refers to Marinduque inoculants; All- means the combination of all treatments)

(a)

(b)

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■ so Days o.o2 0.14 0.01 o.o2 o.o2

■ S0 Days o.o7 o.o5 o.os o.os 0.17

Fig. 9. Diameter increment of T pandacaqui in response to inoculation of amendments when planting (a) with and (b) without A.

auricuiiformis

Based on the results, the application of MykoVAM has standout among other treatments; a fungi-based bio-fertilizer developed by UPLB- BIOTECH. It contains spores, infected roots, and propagules of endomycorrhizal fungi that are estimated to replace 60%-85% of plants' chemical fertilizer requirement and absorb water and nutrients, particularly phosphorus. Also, the inoculants aid the plant in the resistance of pathogens in the roots and increase drought and heavy metal tolerance (de la Cruz, 2012). Phosphorus is vital to the growth of new tissue and the division of cells. It provides the plant the ability to resist disease due to stimulating the fast growth and development of plants (Tajer, 2016). Similar to this amendment was Marinduque inoculant. However, it did not perform well like the MykoVAM. It may be possible that the spores that can be found in the Marinduque soil that were collected had reduced due to the long storage before it was isolated.

Moreover, it was also observed that Bio-N provided a positive impact on the growth of T pandacaqui. Bio-N is a microbial-based fertilizer that converts nitrogen gas into its available form and reduces the nitrogen requirements of crops up to 50%. It was developed by Dr. Mercedes Umali-Garcia and Teofila S. J. Santos of the National Institute of Molecular Biology and Biotechnology (BIOTECH), University of the Philippines Los Baños (UPLB), in 1985 to lessen the cost of fertilizers (Calibo, n.d.). Nitrogen is a substantial food for plants that promotes the vegetative part's growth and development and prompts the uptake and utilization of nutrients, including potassium, phosphorous, and controls overall plant growth (Leghari et al., 2016).

15 replicates out of 50 who survived in the experimental setup for T pandacaqui with A. auriculfformis and only 5 out of 50 were left in the other set up. Length of the roots and number of leaves were higher in T pandacaqui with A. auriculfformis (Fig. 10 & 11). Root colonization of the ten root samples of T pandacaqui without A. auricuiiformis showed 33% of colonization. However, T pandacaqui with A. auricuifformis exhibited higher root colonization having 45% infection. Only the hyphae were accounted for root colonization because other parts of the roots were disfigured during the staining.

5.00 4.50 4.00 3.50 3.00 2.50 2.00 1.50 1.00 0.50 0.00

3.61

1.64 1 1

T. pandacaqui without A. auriculiformis T. pandacaqui with A. auriculiformis

Experimental Set up

Fig. 10. Root development of T pandacaqui in response to planting with and without A. auriculiformis (n=20).

16 14 12 10 8 6 4 2 0

1

11

14

T. pandacaqui without A. auriculiformis T. pandacaqui withA. auriculiformis

Experimental Set up

Fig. 11. Foliage count of T pandacaqui in response to planting with and without A. auricuifformss (n=20).

The growth performance of the indigenous species, T pandacaqui, in terms of root and leaf development was better with A. auricuiiformis. The introduced species' ability to modify soil composition may have aided the growth of T pandacaqui in the copper-rich soil that came from the former mine site in Brgy. Capayang, Marinduque. A. auricuiiformis has a symbiosis in rhizobia and can easily enteran n easily enter relation with new rhizobia species, leading to increased nitrogen content in soil. The association of A. auricuiiformis with both ecto- and endo-mycorrhizal fungi may have influenced the percentage of root colonization of T pandacaqui. A. auriculfformis has a symbiotic relationship with rhizobial bacteria because it provides nitrogen for the species to grow on low nitrogen soils (Duponnois & Planchette, 2003). Treatments with high root colonization were microbial- and fungi-based fertilizers. The species can also significantly raise the soil organic carbon and the cation exchange capacities (Schmerbeck & Naudiyal, 2014). Several countries are using A. auriculiformis. for phytoremediation. It's popular for revegetating mine spoils due to its rapid growth and regeneration that result in abundant litter fall, improving the physio-chemical properties of soilin abundant litter fall, improving the of soilin abundant litter fall, which improvesin abundant litter fall improving soil's physio-chemical properties. The species' phyllodes provide good, long-lasting mulch (Orwa eta., 2009).

Conclusion

Acaulospora species were prevalent from sampled rhizosphere across vegetation types compared with Glomus and Gigaspora, but when mass-produced Acaulospora and Glomus prevailed as an inoculant for T pandacaqui. Further improvement of endomycorrhizal production is necessary to ensure soil inoculant quality for T pandcaqui and other indigenous plants. The height and diameter of T pandacaqui were significant with A. auriculfformis brought by its N-fixing property, though host mortality was higher due to competition. Soil inoculants promoted the growth of T pandacaqui, but MykoVAM performed significantly in height than control. A. auricuiiformis could be maintained as a nurse crop for threatened species like T pandacaqui and consider the potential of local endomycorrhizal isolates to encourage growth and survival.

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Citation:

Belarmino, A.J.D., Pampolina, N.M., Coracero, E.E. (2021). Growth Response of Tabernaemontana pandacaqui Poir. in Soil from a Former Mining Site in Marinduque, Philippines Ukrainian Journal of Ecology, 11 (2), 111-121.

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