The effect of coffee grounds and sawdust Tectona grandis L. f. as planting media for cultivation oyster mushroom Pleurotus sp.

This study aims to determine the effect of the use of coffee grounds and sawdust Tectona grandis L. F as planting media on the cultivation of oyster mushrooms Pleurotus sp. This research was conducted in December 2021 - March 2022. This study used a completely randomized design (CRD) method consisting of 1 factorial. This study used 5 treatments with 3 replications, so a total of 15 baglogs were used. The data obtained were analyzed statistically using the F test and further tested with the 5% Least Significant Difference (LSD). The results showed that the addition of several doses of coffee grounds had a significant effect on fruit body growth time, fruit body fresh weight, fruit body dry weight, and fruit cap diameter. The fastest growing time of oyster mushroom mycelium was at P0 (sawdust Tectona grandis 2 kg + control) an average of 4.67 days, the fastest fruit body formation speed was P0 (sawdust Tectona grandis 2 kg + control) with an average of 15.67 days, fresh fruit weight the fastest with an average of 106.33 g, the dry weight of the fruit body of the heaviest oyster mushroom with an average of 33 g, and the diameter of the largest fruit hood with an average of 30.3 cm, meanwhile for the P3 treatment (sawdust Tectona grandis 0.5 kg + 1.5 kg coffee grounds) and P4 (coffee grounds 2 kg + control) were not suitable for the growth of oyster mushrooms.


Introduction
Cultivation of oyster mushroom is one of the agribusiness businesses that has quite a big business opportunity because in the last 10 years its economic value has continued to increase.Oyster mushrooms are a type of edible mushroom that belongs to the Genus Pleurotus and the family Pleurotaceae [1,2].Pleurotus is a genus of white-rot basidiomycete fungi known for their complex enzymatic system and prominent lignocellulolytic properties [3][4][5].Oyster mushroom is a type of wood mushroom that can be consumed [6][7][8], The nutritional content of oyster mushrooms is 19-35% protein, 9 amino acids, 72% unsaturated fat and fiber content from 7.4 to 24.6 [4,[9][10][11][12].Oyster mushroom is widely cultivated for its medicinal properties as well as its delicacy [13][14][15][16], this makes oyster mushroom cultivation a good business opportunity both on a small or large scale [14,17].
Oyster mushroom is known as a mushroom that is easy to cultivate and is widely developed on wood substrate media that has been packaged in plastic bags and then incubated and maintained [11,18].Media is one of the environmental factors that functions to provide nutrients and water for plantlet growth [19].Oyster mushrooms will grow in nutrient-rich media [3].Mushroom cultivation uses sawdust as a raw material for making baglog for mushroom nurseries due to the presence of cellulose, 1230 (2023) 012071 IOP Publishing doi:10.1088/1755-1315/1230/1/012071 2 hemicellulose and lignin content [7,[20][21][22].Sawdust can stimulate the growth of the mushroom mycelium faster on the media [8].The best oyster mushroom growing media comes from sawdust of broadleaf trees and is classified as hardwood because it does not contain much sap [9,18,23].Hardwood sawdust produced better quality oyster mushroom nutrition than softwood sawdust [24].
Tectona grandis L. f is a type of broadleaf tree and includes broadleaf trees and hardwoods, Tectona grandis sawdust contains carbohydrates, cellulose, hemicellulose and lignin which can be decomposed and can also be used as a growth medium for oyster mushrooms [11,25].However, some mushroom farmers are currently having difficulty locating sawdust [6,26].This happens because the potential of the forest is currently reduced and limited.Efforts to anticipate this need to find alternative substrates that are available and easy to obtain as oyster mushroom cultivation media [26,27], alternative materials that can be used to replace sawdust, one of which is coffee grounds [1,18,28].Coffee wastes are good substrates for fungus growth, which can be used to produce Oyster mushrooms Pleurotus sp.[15,29].Coffee grounds waste contains 1.2% nitrogen, 0.2% phosphorus and 0.35% potassium needed for growth and can be used as an additive in oyster mushroom growing media.[30].
Cultivating oyster mushrooms using sawdust and coffee grounds is a waste management that provides added economic value, restores the environment and maintains community food security [2,15,28].Therefore, based on the above background, a study was conducted on the effect of using coffee grounds and sawdust of Tectona grandis L. f as a planting medium for Pleurotus sp.oyster mushroom cultivation.

Materials and Methods
The research was conducted in December 2021 -March 2022 at the Botanical Laboratory, Department of Biology, Faculty of Mathematics and Natural Sciences, Hasanuddin University.

1 Preparation and Mixing of Planting Media
The coffee grounds obtained from the rest of drinking coffee are dried manually in the sun.The research design used was Completely Randomized Design (CRD).This study used 5 (five) treatments with 3 (three) replications, so that a total of 15 baglogs were used.Each media composition was added 240 g rice bran, 20 g agricultural lime CaCO3 to maintain pH balance [16,30], sufficient water was estimated so that the media did not drip water when held and did not break.The treatment was the addition of coffee grounds and Tectona grandis sawdust with different concentrations in the planting medium.These treatments are: ▪ P0 : Tectona grandis sawdust 2 kg + coffee grounds 0 kg (control) ▪ P1 : Tectona grandis sawdust 1.5 kg + coffee grounds 0.5 kg ▪ P2 : Tectona grandis sawdust 1 kg + coffee grounds 1 kg ▪ P3 : Tectona grandis sawdust 0.5 kg + coffee grounds 1.5 kg ▪ P4 : Tectona grandis sawdust 0 kg + coffee grounds 2 kg (control)

2 Packaging and Sterilization
The media is wrapped in a PP plastic bag (called a baglog) and sterilized using a sterilization chamber (drum) for 8 hours, then cooled for ± 24 hours to an indoor temperature of 35 -40°C.

Inoculation and Incubation
Inoculation is done by taking some of the mushroom seeds using a sterile spatula aseptically and inoculating them into new planting media in a baglog.Baglog was covered with HVS paper and tied with a rubber band, then incubated in the incubation room.Incubation was carried out at temperatures ranging from 22 -28ºC with 60-70% humidity [10,32,33].The baglog that has been filled with mycelium (30-40 days after inoculation) is then transferred to the mushroom house for the growth of the fruit body (pin head) with a temperature of 16-22ºC and 80-90% humidity [16,33].

Enlargement of Oyster Mushroom Fruit Bodies
After incubation at room temperature 22 -28ºC until the entire media is full of white mushroom mycelium, for 3 -4 weeks the baglog cover is opened so that the mushroom fruiting bodies can grow.The conditions required for the growth of fruiting bodies are temperatures of 16 -28ºC [10,34].

Harvesting Oyster Mushrooms
Harvesting is done when the size is large enough, around 5-10 cm.Harvesting is done by removing all of the existing mushroom clumps, until no part of the mushroom is left on the baglog media.The harvested mushrooms are then cleaned, and the lower part of the stem is cut.After the 1 st harvest, the wet weight of the oyster mushrooms was weighed in the baglog for each treatment.Baglog that has been harvested, the back plastic is torn with a razor blade so that the next fruiting body can emerge from the back of the baglog.

Observed parameters
Good media composition for the growth of oyster mushrooms consists of sawdust, lime and rice bran.
Observations were made once a day, starting from the first day after inoculation until harvest.
Parameters measured were: a) Mycelium growing time, b) Fruit body growing time, c) Fruit body wet weight, d) Fruit body dry weight, and e) Cap diameter [33].

Research Data Analysis
The research data were analyzed using ANOVA at the 95% level of confidence to determine the effect on the treatment, if the ANOVA showed a significant difference, then a further test would be carried out with the Least Significant Difference (LSD) test [35] 3. Results and Discussion

Effect of adding coffee grounds on mycelium growth time
The results of research conducted by providing basic media with the addition of coffee grounds with different concentrations showed that there were differences in the speed of mycelium formation at various ages of observation.This research was conducted in a dark room with temperatures ranging from 25-28ºC, incubation was carried out by storing media filled with seeds under certain conditions so that the mushroom mycelium could grow.The temperature required for mycelial growth is between 22-28ºC [36].Mushroom cultivation requires temperature and humidity control to obtain the best fungal growth [32].The results of the analysis of variance showed that the treatment with coffee grounds did not significantly affect the mycelium growth time, so further tests were not necessary (Table 1).4.67 a P1 : Tectona grandis sawdust 1.5 kg + coffee grounds 0.5 kg 5.00 a P2 : Tectona grandis sawdust 1 kg + coffee grounds 1 kg 5.00 a P3 : Tectona grandis sawdust 0.5 kg + coffee grounds 1.5 kg 5.67 a P4 : Tectona grandis sawdust 0 kg + coffee grounds 2 kg (control) 6.33 a Note: Treatments that have different notations in one column mean they have a real effect.
In treatment P0 (Tectona grandis sawdust 2 kg) had the fastest mycelium growth which required an average incubation time of 4.67 days, while the slowest mycelium growth was treatment P4 (2 kg coffee grounds) as a control for coffee media requiring the longest average incubation time was 6.33 days.Determination of mycelial biomass from 7 day colonies [4].Mycelium growth is calculated from the time the baglog is inoculated until the mycelium has completely filled the baglog [27].The environmental conditions of the incubation site also influence the incubation period of mycelium [37,38].
The thickness of the mycelium is obtained by observing the mycelium that meets the treatment baglog.The thickness of the mycelium for all treatments was different, the more the concentration of coffee grounds, the thicker the mycelium and did not fill the entire baglog.The P3 (Tectona grandis sawdust 0.5 kg + coffee grounds 1.5 kg) and P4 (Coffee grounds 2 kg) treatments were low because the dose of coffee grounds used was too much, this caused the baglog media to become dense and filled the entire baglog, unable to spread and the mycelium is very thin in the form of thin white threads (Figure 1).The more concentration of coffee grounds is given, the longer it takes for mycelium to grow [10], this is because the pH of coffee grounds is too high, namely 6.2.pH concentration is an important factor for oyster mushroom production [16,26].Optimal fungal growth at pH 6 -8 and for mycelium growth at pH 8 [16,39].

Effect of Addition of Coffee Grounds on Growing Time of Fruit Bodies
The results of the analysis of variance showed that the coffee grounds had a significant effect on the formation of fruiting bodies, so further tests were needed.This is based on a significance value that is smaller than α 0.05 (Table 2).15.67 b P1 : Tectona grandis sawdust 1.5 kg + coffee grounds 0.5 kg 20 b P2 : Tectona grandis sawdust 1 kg + coffee grounds 1 kg 18.5 b P3 : Tectona grandis sawdust 0.5 kg + coffee grounds 1.5 kg 0 a P4 : P4 : Tectona grandis sawdust 0 kg + coffee grounds 2 kg (control) 0 a There is a real influence in the provision of media with the addition of coffee grounds due to the addition of nutrients to the mushroom growing media which can increase the yields obtained [39].Based on data on the average length of emergence of shoots formed (Table 2) treatment P0 produced the fastest average duration of emergence, namely 15.67 days, whereas in treatments P3 and P4 no shoots and fruiting bodies appeared because the mycelium growth time was also the slowest, because the speed at which the mushroom fruiting bodies appear is strongly influenced by the growth of the mycelium [31,40].The growth of shoots and fruit requires air humidity ranging from 80-85% [16], environmental and nutritional factors affect how long shoots appear [34,40].Coffee grounds are made up of a lot of water and organic material, the organic material includes things like lignin, cellulose and hemicellulose [5,11,28].The reduced formation of fruiting bodies is due to the fact that some of the nutrients needed have been used for the growth of the mycelium, so that not all of the primordials that grow become fruiting bodies [4,26,40,41].The C/N ratio had more effects on the mycelium growth, the formation and development of fruiting bodies [5,33,42,43].The addition of calcium to produce large fruit bodies [11].

Effect of adding coffee grounds to fruit fresh weight
The results of the analysis of variance showed that the treatment with coffee grounds had a significant effect on the fresh fruit weight, so further tests were needed.Based on data on the average fresh weight of the fruit bodies formed (Table 3), there are differences in response to each treatment.Treatments Average (g) P0 : Tectona grandis sawdust 2 kg + coffee grounds 0 kg (control) 106.33 b P1 : Tectona grandis sawdust 1.5 kg + coffee grounds 0.5 kg 69.33 b P2 : Tectona grandis sawdust 1 kg + coffee grounds 1 kg 29.66 b P3 : Tectona grandis sawdust 0.5 kg + coffee grounds 1.5 kg 0 a P4 : Tectona grandis sawdust 0 kg + coffee grounds 2 kg (control) 0 a Mushroom fresh weight is used as a growth parameter because it can be measured directly.Measurement of the fresh weight of oyster mushrooms is when after harvest it is directly measured using a scale.Treatment P0 produced the highest average fresh fruit weight, namely 106.33 g, treatment P1 (69.33 g), and treatment P2 (29.66 g), while treatments P3 and P4 (Table 3) did not produce oyster mushrooms.So, there is no fruit weight.This is because the media in the baglog is very dense so that the mycelium cannot grow properly as a result, the growth of the fruit bodies is also hampered [40].The addition of nutrients to the media affects the number of fruit bodies formed and fresh weight of fruit bodies [26,43].The weight of oyster mushrooms can be influenced by the number of primordial mushrooms [40].

Effect of adding coffee grounds to the dry weight of fruit bodies
The results of the analysis of variance showed that the treatment of coffee grounds had a significant effect on the diameter of the fruit cap.This is based on a significance value that is smaller than α 0.05 (Table 4).Oyster mushroom has a highwater content as evidenced by holding the oyster mushroom.Almost all types of fresh mushrooms have a water content of 85-95%, while dried mushrooms contain only 5-20%.
The results of the calculation of the average dry weight of fruit bodies were greatest in treatment P0 of 33 g, while the results of the calculation of the smallest average dry weight were P2 of 16.5 g (Table 4).Even though sawdust treatment P0 had a higher dry weight, the results were not much different from treatment P1 which was given coffee grounds.This is possible because the phosphorus content also contributes to the dry weight of oyster mushrooms [26].The dry weight is the overall weight of the nutrients apart from the water content in the oyster mushroom fruit.The determining factor for the high and low productivity of white oyster mushrooms is the weight of the fruit bodies [11].In the P0 treatment it is possible that the fat and crude fiber content in the fruit bodies is relatively high.This is because the content of fat and fiber in baglog P0 is relatively the highest when compared to other treatments.Corganic is a source of energy for maximum mycelium and mushroom cap growth [43].

The Effect of Adding Coffee Dregs on the Diameter of the Fruit Cap
The results of the analysis of variance showed that the treatment of coffee grounds had a significant effect on the diameter of the fruit cap.This is based on a significance value that is smaller than α 0.05 (Table 5).Based on the data obtained, the average hood diameter formed in Table 5 shows that there are differences in response to each treatment.The P0 treatment produced the highest average fruit cap diameter, namely 30.3 cm, while the P2 treatment produced 5.07 cm.The P0, P1, and P2 treatments had faster mycelium growth, mycelium growth was the initial stage of fruit body formation.Treatments P3 and P4 with the highest dose of coffee grounds did not grow fruit bodies due to decay, which meant that no fruit caps were formed.Decay occurs because the mycelium does not grow optimally because the media used is too dense, because too much coffee grounds are used, this makes it difficult for the mycelium to spread throughout the baglog media.The mycelium growth of oyster mushrooms was improved when the carbon source was glucose, molasses, or a combination of both (1-5%) [36].Environmental factors that affect the growth of oyster mushrooms are temperature, humidity, light intensity and air circulation [11,39]  The diameter of the fruit cap is strongly influenced by the number of fruit bodies and the availability of nutrients in the media, because there are more fruit bodies [43].When many fruiting bodies have grown, a little nutrition is distributed to each fruiting body because the nutrients needed have not been decomposed properly, plus the number of fruiting bodies that are formed makes each fruiting body only get nitrogen which affects the area of the cap [5,11,41].The nitrogen used in plant growth comes from sawdust materials [40].

Conclusion
From the research results obtained, it can be concluded that the addition of several doses of coffee grounds had a significant effect on the growth time of fruit bodies, fruit body fresh weight, fruit body dry weight, and fruit cap diameter.The fastest growing time for oyster mushroom mycelium was at P0 an average of 4.67 days, the fastest speed of formation of oyster mushroom fruit bodies was P0 with an average of 15.67 days, the fastest fresh weight of fruit bodies with an average of 106.33 g, The heaviest oyster mushroom fruit body with an average of 33 g, and the largest fruit cap diameter with an average of 30.3 cm, meanwhile the P3 and P4 treatments were not suitable for the growth of oyster mushrooms.

Tabel 3 .
Average Fresh Weight of Oyster Mushroom Pleurotus sp. of Various Treatments

Figure 2 .
Figure 2. The largest fruit cap diameter was obtained in baglog P2

Table 1 .
Mycelium Growth Time in Pleurotus sp.Oyster Mushroom Growth Media. of Various

Table 2 .
Average Growth Time of Fruit Bodies Growth Media for Oyster Mushroom Pleurotus sp. of

Table 5 .
Average Diameter of the Oyster Fungus Pleurotus sp. of Various Treatments