Propagation of mycorrhiza as biological fertilizer and its application on patchouli seedling in Andisols Bener Meriah

Mycorrhiza, functioning as a biofertilizer, have the capacity to improve soil fertility, including Andisols which is cultivated by patchouli. This research objective to produce mycorrhizal as a biofertilizer to increase the growth of patchouli. Additionally, the effect of mycorrhizal types on the seedlings of patchouli in Andisols was studied. Propagation mycorrhizae carried out the exploration of mycorrhizae on Andisols Bener Meriah, and the carrier was Glomus mosseae, Gigaspora, and a mixture of Glomus mosseae and Gigaspora sp. and then the mycorrhizal biofertilizer was tested to patchouli seedling. The parameters observed in this study included the colonization of patchouli root, initial soil analysis, and patchouli seedling growth. The output in this study is a product in the form of a local specific strain of Bener Meriah biofertilizer. The results obtained have succeeded in the propagation mycorrhizal biofertilizers with the carrier Glomus mosseae, Gigaspora, and a mixture of Glomus mosseae and Gigaspora sp. The highest 90% mycorrhizal colonization was found in the mixed mycorrhizal carrier Glomus mosseae + Gigaspora sp. The growth of patchouli seedlings increased, and the highest patchouli growth was observed the combination of mycorrhizal Glomus mosseae and Gigaspora sp. application


Introduction
Pogostemon cablin Benth., also referred to as patchouli, is recognized as an important plantation commodity that is extensively cultivated by various communities.Patchouli is a producer of essential oils and is important to the bio-pharmaceutical, cosmetics, and perfume industries.Aceh supply around 90% of Indonesia's patchouli oil production to the world.In addition to its application as a component in perfume production, Aceh patchouli possesses several beneficial properties for aromatherapy, the treatment of diabetic wounds, skin smoothing, anti-aging effects, prevention of hair loss, as well as antibacterial and disinfecting properties.Patchouli is widely recognized as an important plantation crop that plays a substantial role in fostering the economic advancement of the local population.Furthermore, it is worth noting that patchouli exhibits promising potential and serves as a key agricultural commodity in many regions of Aceh, including Bener Meriah, Aceh Jaya, North Aceh, South Aceh, and West Aceh.The genetic and environmental factors impose an important effect on both the yield and quality of patchouli.Organic patchouli is essential for the perfume, cosmetics, and biopharmaceutical sectors because of its enhanced safety and health benefits.Moreover, the extraction of patchouli oil from organically cultivated patchouli yields a notably high grade [1].1297 (2024) 012022 IOP Publishing doi:10.1088/1755-1315/1297/1/012022 The variations production of patchouli can be attributed to environmental factors, including both the chemical and physical properties of the soil, climatic condition, and land physical characteristics such as altitude, slope, and surface rocks [10].The cultivation of organic patchouli needs a special emphasis on the chemical and biological characteristics of the soil.One additional concern associated with certain patchouli plant cultivars is their vulnerability to disease infestations.In this regard, mycorrhizae can serve as a bioprotective agent, hence enhancing patchouli yield.The utilization of mycorrhizal biological fertilizers has the ability to enhance the chemical, physical, and biological properties of Andisols, hence promoting fertilization.Mycorrhizal fungi demonstrate efficacy in enhancing nutrient uptake in nutrientdeficient (marginal) soil by facilitating the absorption of essential elements such as phosphorus (P), nitrogen (N), and potassium (K), Consequently they are available to plants.The existence of mycorrhizae is essential for the accessibility of nitrogen (N), phosphorus (P), and potassium (K) nutrients in the soil, especially in Andisols [8].
The cultivation of patchouli in Aceh is commonly practiced on Andisols, Ultisols, Inceptisols, and Entisols by local farmers.Patchouli cultivation in the highland regions, such as Bener Meriah, Gayo Lues, and Aceh Tengah, primarily is carried out in Andisols.Andisols, a type of soil characterized by high fertility and volcanic ash content, were found to occupy around 5.4 million hectares, accounting for 2.9% of Indonesia's overall land area.[2].The soil formation process is attributed to the deposition and weathering of volcanic debris.The distinguishing features of Andisols encompass a notable presence of organic carbon, low bulk density, high phosphorus retention, and cation exchange capacity (CEC) [3].One notable attribute of Andisols is their high concentration of amorphous or allophane clay minerals, with a prominent presence of aluminum (Al) and iron (Fe).Due to the substantial presence of organic matter in Andisols, these soils exhibit a high degree of suitability for the proliferation and growth of arbuscular mycorrhizal fungi (AMF) [4].
AMF have the ability to contribute to plant nutrition and pathogen suppression in low-input agricultural systems is a promising perspective in modern agriculture [5].However, it is important to note that the functional properties of AMF can vary significantly among different species.Recent research has indicated that the utilization of AMF inoculum is highly suggested in certain agricultural systems as a beneficial practice for the management of organic farms.Yet, there is a scarcity of knowledge about the tolerance of patchouli in response to the application of organic fertilizers and mycorrhizal biofertilizers on Andisols.In order to address this objective, it is necessary to conduct a research investigation pertaining to the cultivation of organic patchouli utilizing mycorrhizal biofertilizers on Andisols.Multiple investigations have yielded findings that demonstrate enhanced growth of patchouli treated with mycorrhizae.In addition to this, the issue pertaining to the growth and yield of patchouli in drought conditions might potentially be mitigated through the utilization of mycorrhizal biofertilizers.The presence of mycorrhizae in the roots has been shown to enhance the capacity of plants to withstand the adverse effects of drought stress.In general, the highest concentration of leaf alcohol in patchouli was observed when mycorrhizal application treatments were combined with high drought stress condition [6].
Several papers provide confirmation of the mycorrhizal inoculation of patchouli utilizing various strains.The selection of an appropriate strain of mycorrhiza is a crucial factor in determining the yield of patchouli, particularly in the context of organic patchouli production.The various AMF for symbiotic interaction, including Glomus etunicatum, has been determined as the most effective mycorrhizal symbionts for enhancing the growth and uptake of phosphorus (P) nutrients in patchouli [7].The yield of plants, such as patchouli, is strongly influenced by the specific type of mycorrhizae present.The aforementioned phenomenon might be attributed to the capacity of mycorrhizal colonized root to uptake nutrients and provide protection to plants against diseases, drought stress, and various other adverse environmental situations [8], assisting in the phosphate and nitrogen uptake [4] and producing hormones, for example auxin and gibberellin [9].
Based on previous explanation, it is necessary to study the mycorrhizal propagation technique and its utilization to improve the growth and production of patchouli seedling on Andisols Bener Meriah.

Material and Method
This study focused on the propagation technique the biofertilizer mycorrhiza on Andisols and testing the fertilizer for the patchouli seedlings growth in greenhouse pots.The materials and tools used in this study were the Lhokseumawe patchouli variety, the AMF inoculum of Glomus mosseae and Gigaspora species and a mixture of both with zeolite rock carrier, the fertilizers urea, SP-36 and KCl applied a half of the recommended dose, maize seed, zeolite, hand tractor, analytical scale, sprayer, oven, sieve, bucket, paint, board, A4 envelope, stereo microscope, scale, wood, plastic sack, A4 envelope, and microscope.
The experiment to produce mycorrhizal biofertilizer on Andisols for the treatment of organic patchouli.The host plant Zea mays was utilized for the propagation of mycorrhiza.The propagation process involves the implementation of plant management techniques such as topping and stressing.The Mycorrhizal inoculum was collected from the host plant 75 days post-planting.The experiment of the application mycorrhiza biofertilizer on patchouli seedling was a pot experiment and conducted in the greenhouse.The design used in this study was a non-factorial randomized block design with three replications.Mycorrhiza (M) factor consisted of three levels, namely: (a) M1 = Glomus moseae, (b) M2 = Gigaspora sp, and (c) M3 = Combined Glomus+Gigaspora.The parameters in this study were the initial soil analysis (before treatment) including soil texture (Pipette method), soil moisture content, pH H2O 1: 2.5 (electrometry method), total N (Kjeldahl method), P available (Bray I), KTK (Atomic Absorption Spectrophotometer (AAS) Method).The observed variables are pH H2O 1: 2.5 (electrometric method), N-total (Kjeldahl method), available P (Bray I method) and CEC (AAS method).Analyzes were performed for N (Kjeldahl method), and P (Spectrophotometry method).The patchouli root mycorrhizal colonization or the degree of AMF infection was observed using staining method according to [11].
The observed parameters of growth patchouli seedling were plant height, stem diameter, total leaves, and total branches.The data of the growth of patchouli were analyzed with analysis of variance and Tukey's HSD test at the 5% level.

Root infection of mycorrhizal colonization
The experiment has been successfully conducted, resulting in the production of mycorrhizal biofertilizer derived from three different species of fungi: Glomus moseae, Gigaspora sp., and a mixture of Glomus moseae and Gigaspora, sp.Table 1 shows the degree of root infection by mycorrhizal biofertilizer.Based on the data presented in Table 1, it can be observed that the combination inoculation of Glomus mosseae and Gigaspora sp.resulted in the maximum degree of mycorrhizal infection.This very high infection can be attributed to the fact that the combined treatment of Glomus mosseae and Gigaspora sp resulted in a greater production of root and canopy biomass, as compared to the separate treatments of Glomus mosseae and Gigaspora sp.treatment.According to the findings of [12], it has been observed that Glomus sp mycorrhizae demonstrate favorable adaptability and growth in both polluted and tropical environments.The utilization of mycorrhizal starters, specifically Glomus sp and Gigaspora sp, in experiments involving the maize host plant exhibited a notable disparity in root biomass.The mycorrhizal starter Glomus sp yielded significantly higher root biomass compared to the starter Gigaspora sp.Furthermore, when the two starters were combined, a synergistic effect was observed, leading to enhanced stimulation of plant development.Moreover, a study conducted by [13] revealed that the root growth of chili was significantly enhanced when treated with a combination of Glomus mosseae and Gigaspora sp biofertilizer, as compared to the individual application of Glomus mosseae or Gigaspora sp.The study conducted by [14] revealed that the utilization of a mixed mycorrhizal starter consisting of Glomus mosseae and Gigaspora sp led to a significant increase in root biomass when compared to the individual utilization of Glomus mosseae or Gigaspora sp.The application of mycorrhizal biofertilizer has demonstrated significant efficacy in promoting the growth of organic patchouli seedlings.
The achievement of mycorrhizal propagation is influenced not only by the type starter of mycorrhiza, but also by the environment condition during mycorrhizal biofertilizer propagation.The propagation of mycorrhizal fungi is influenced by environmental conditions, which have been observed to result in a significant increase in root infection.This is particularly evident in the species Glomus moseae, Gigaspora sp, and Mixed species.).The study conducted by [15] demonstrated that the success rate of mycorrhizal propagation is influenced by various environment factors, including soil pH, temperature, humidity, and soil type.Moreover, [16] noted that the efficacy of mycorrhizal infection is strongly influenced by the carrier type of mycorrhizal biofertilizers.Moreover, the utilization of a zeolite carrier has been shown to enhance the development and productivity of maize plants.In a prior study conducted by [6], it was observed that the application of mycorrhizal biofertilizer led to enhanced growth, as well as increased viability and vigor of patchouli seedlings.)

Initial Soil Analysis
Table 2 shows the initial soil analysis conducted at the research site.Based on the preliminary soil analysis, it can be inferred that the soil fertility level of Andisols Bener Meriah is categorized as low.According to the analysis conducted, it has been determined that the Andisols Bener Meriah has a texture of loam.The preliminary assessment of Andisols Bener Meriah indicated that the soil exhibited a classification of very strongly acidic (pH KCl) and strongly acidic soil reaction (pH H2O).The problematic is that the Andisols Bener Meriah P soil is immobile form (fixed) and hardly available for plants.Although the organic matter content in the soil is classified as high level, the total N is classified as medium category, and the available-P is classified as low level and the soil texture contain slightly high of sand.The application of mycorrhizae, it is expected that the availability of nutrients in soil and the nutrient uptake by plant will be high and can increase the growth and yield of patchouli in Andisols Bener Meriah.In general, there is a correlation between AMF and the increase of soil pH value and affect soil fertility.This is due to the activity and metabolism of AMF which can increase the release of organic compounds in binding metal cations that play a role in increasing soil pH.[17] argues that these organic compounds in the sorption complex are able to bind cations.This causes the base saturation of the soil is high as soil pH increased.Based on the results of the preliminary analysis, it appears that the Andisols Bener Meriah has a very low P availability content.Therefore, the absorption of P is expected to be optimal with the use of mycorrhizal bio-fertilizers in addition to other nutrients.Several studies have shown that P is a major limiting factor in Ultisols and Andisols.Low and P bound in Andisols cause plants including patchouli cannot absorb it effectively.Besides that, the type of mycorrhiza applied greatly determines the effectiveness of P absorption [8,18].In detail, the results of the initial analysis of Andisols Bener Meriah are shown in Table 2.

Patchouli Seedling Parameters
The results of mycorrhiza applications on the growth parameter of patchouli are recorded in Table 3-6.The data shown in Tables 3, 4, 5 and 6 demonstrate that application of mixed mycorrhiza Glomus mosseae + Gigaspora sp showed the significant different effect on the growth of the patchouli plant and exhibits a notable improvement when exposed to mixed mycorrhizal strains, as evidenced by increases in plant height, plant diameter, number of leaves, and number of branches.The results of this study confirmed that the mixed mycorrhizae strain had superior seedling growth yields in comparison to the other two strains.
One of the key roles of mycorrhizae in plants includes facilitating the uptake of nutrients from the soil, thereby promoting plant growth.Additionally, mycorrhizae serve as a biological defense mechanism against pathogenic infections in plant roots.Moreover, they enhance water availability for plants and stimulate the production of growth-promoting hormones [19].Mycorrhizal biological fertilizers have the capacity to enhance the nutrient uptake in less fertile soils, hence facilitating the absorption of fixed phosphorus (P) nutrients that subsequently become accessible to plants.The medium level of nitrogen in the media as shown in the laboratory analysis of Andisols Bener Meriah also enhance the availability of nitrogen to plant by mycorrhiza, increase in nitrogen levels at a low to medium range has been found to enhance arbuscular mycorrhizal (AM) colonization and sporulation, as well as promote plant growth and root formation.The provision of nitrogen throughout the early stages of development has been seen to potentially confer advantages in the creation of mycorrhizal associations [20].

Conclusion
The propagation technique of mycorrhizal biofertilizer has been conducted and the best strains was observed in the combined of Glomus mosseae and Gigaspora sp application with a root colonization degree of 90% (classified as very high).Provision of mycorrhizal biofertilizer increases patchouli seedlings growth, and the highest patchouli growth was observed the combination of mycorrhizal Glomus mosseae and Gigaspora sp.application.

Table 1 .
Types of mycorrhizal biofertilizers and degree of root infection.

Table 2 .
Data analysis of soil properties before treatment (initial).

Table 3 .
The average height of patchouli seedling at 15, 30 and 45 days after planting (DAP) due to the application of mycorrhiza on Andisols Bener Meriah.

Table 4 .
The average leaves of patchouli seedling at 15, 30 and 45 days after planting (DAP) due to the application of mycorrhiza on Andisols Bener Meriah.

Table 5 .
The average stem diameter of patchouli seedling at 15, 30 and 45 days after planting (DAP) due to the application of mycorrhiza on Andisols Bener Meriah

Table 6 .
The average branches of patchouli seedling at 15, 30 and 45 days after planting (DAP) due to the application of mycorrhiza on Andisols Bener Meriah