The Role of Bacteria Isolated from Different Sources in Melon Cultivation: Effects on Yield and Quality

doi:10.21203/rs.3.rs-4802061/v1

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The Role of Bacteria Isolated from Different Sources in Melon Cultivation: Effects on Yield and Quality

https://doi.org/10.21203/rs.3.rs-4802061/v1

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This research was conducted to determine the effect of bacterial strains isolated from different sources on the development of melon plants. Plant growth promotion mechanisms such as calcium, potassium and phosphorus solubilization, nitrogen fixation, phytohormone, siderophore and ACC-deaminase production and their growth at different pH and salt concentrations were determined. In order to determine the effect of bacterial strains on plant growth, field trials were established according to the randomized blocks experimental design with three replicates and carried out with two years of repetition. In the experiment established for this purpose, 11 different applications [IT 22 (Bacillus safensis), IT 22 + Fertilizer, IT 63 (Acinetobacter calcoaceticus), IT 63 + Fertilizer, IT 93 (Acinetobacter calcoaceticus), IT 93 + Fertilizer, IT 115 (Serratia rubidaea), IT 115 + Fertilizer and control (fertilizer only)] were included. In terms of all parameters examined, the best result was obtained from the Mix + Fertilizer application, while only the highest value of WSDM (9.9%) was measured in the IT 93 + Fertilizer application. The results of the study show that fertilizer-free IT 93 and Mix applications provide higher melon yield than fertilizer, suggesting that bacterial single or mixed applications can be used as a very effective method to reduce the use of chemical fertilizers. In addition, higher fruit yield per decare was obtained in the blocks where bacteria and fertilizer were applied together, except for IT 115 + Fertilizer application, compared to the control group. This result was an indication that a significant reduction in chemical use will be achieved with the inclusion of the determined bacterial strains in fertilization programs.

Cucumis melo

Biofertiliser

Sustainable agriculture

PGPB

Yield

Melon, which is cultivated in large areas worldwide and has the potential to increase the income of producers, is a garden plant in the Cucurbitaceae family (Makful et al. 2017). In 2021, global melon production reached 42 million tons, with the majority produced by China. With a production quantity of 1,638,638 tons, Turkey ranked as one of the top melon-producing countries after China (FAO 2021).

Demand for agricultural products is increasing with population growth. Unfortunately, in order to meet this demand, traditional farming systems, which use intensive inputs, rely heavily on chemicals to improve crop yield and fruit quality (Martínez et al. 2019). However, unconscious and excessive use of these chemicals in agricultural production causes pollution of natural resources such as soil and water, decrease in the diversity of microorganisms in the soil, and thus decrease in soil fertility and quality. This situation leads to a decrease in the quality and quantity of agricultural products (Doktor and Bulut 2023). For this reason, in recent years, the development of sustainable production systems targeting the use of organic inputs in agriculture has become important. Studies aimed at producing more and healthier food thanks to reduced chemical input through biotechnological processes are increasing day by day (Bhardwaj et al. 2014). Sustainable or good agricultural practices; It aims to use natural resources such as soil, water and plants effectively and efficiently, to protect the environment, to ensure food safety for public health and, finally, to leave a livable nature to future generations. One of the environmentally friendly alternative methods to be applied at this point is the use of plant growth-promoting bacteria (PGPB). This understanding, which is focused on sustainable agriculture, reveals the necessity of biological applications instead of chemical use in agricultural production (Toprak 2012). PGPBs have a growth-promoting effect in plants through different means, such as increasing nutrient availability (phosphorus and potassium solubility, biological nitrogen fixation, siderophore production) and phytohormone production (Mia et al. 2012). It also increases plant resistance to biotic stress caused by various pathogens through some mechanisms (secretion of antagonistic substances, antibiotic production, etc.) (Khan et al. 2021). However, it has been reported that some PGPBs can help plants resist abiotic stresses, and at this point, their use in agriculture is seen as a new and promising approach to increase the success of remediating contaminated soils (Tak et al. 2013).

In various previous studies, it has been determined that bacteria in the genera Bacillus, Pseudomonas, Serratia, Enterobacter and Streptomyces have a great potential in vegetable cultivation, promote the growth of many plant species and increase yield (Bhattacharyya and Jha 2012; Karpagam and Nagalaleshmi 2014). In this study, it was aimed to determine the plant growth-promoting properties of bacterial strains isolated from different sources in order to reduce the chemicals used extensively in agricultural production, and to reveal the effects of these bacterial strains on some yield and quality parameters of melon grown under field conditions by using them singly and in combination.

In this study conducted to determine the effect of bacterial strains on some yield and quality parameters of melon plant, Sürmeli F1 melon variety, which is the most preferred by producers in Iğdır, was used. In the research, samples were taken from the phyllosphere and rhizosphere regions of some alternative forage plants growing in extreme conditions (Chepodium qinoa, Amaranthus paniculatus, Atriplex nitens and Salsola rutenica) and volcanic rocks collected from Iğdır Taşburun Village, and a total of 189 bacteria were obtained and their effects on plant development were tested. Among these strains, 4 of them [IT 115 strain with antibacterial activity against Acidovorax citrulli (Temel 2023) and IT 22, IT 63 and IT 93 strains with high plant growth promoting properties as a result of the tests] were used as PGPB.

The biochemical properties of the isolated bacterial strains were determined by oxidase, catalase, gram reaction (Narayanasamy 2001), starch hydrolysis (amylase production) and levan colony formation tests (Hélias et al. 2012). Hypersensitive reaction test was used to test whether the strains were pathogenic or not. The strains were also diagnosed by fatty acid methyl ester analysis (Sasser 1990) and 16S rDNA sequence analysis (Ficus Biotechnology). The results of 16S rDNA sequence analysis of the strains were taken as basis in the study. The characteristics determined about the bacteria are presented in Table 1. Images of the tests are given in Fig. 2.

Table 1

Identification results and some characteristics of bacteria used as PGPB
BS*	Identification result (16S rDNA sequence analysis)	O	C	GR	LCF	A	HR	Isolation Material
IT22	Bacillus safensis	-	+	-	+	-	-	Amaranthus paniculatus-stem
IT63	Acinetobacter calcoaceticus	-	+	+	-	-	-	Volcanic rock
IT93	Acinetobacter calcoaceticus	-	+	+	+	-	-	Salsola rutenica- root
IT115	Serratia rubidaea	-	K+	-	+	-	-	Atriplex nitens-soil
*BS: Bacterial strains, O: Oxidase, C: Catalase, GR: Gram Reaction, LCF: Levan Colony Formation, A: Amylase, HR: Hypersensitive Reaction

Climate data for the period when the experiment was conducted were obtained from Iğdır Meteorological 16th Regional Directorate (Table 2).

Table 2

Climate data for the period when the experiment was conducted
Months	Total Rain Fall (mm)			Average Temperature (℃)			Average Relative Humidity (%)
Months	2021	2022	LAG*	2021	2022	LAG	2021	2022	LAG
March	46.4	24.8	21.3	7.4	5.1	7.0	55.3	54.9	50.2
April	18.4	25.8	38.5	17.4	15.7	13.4	44.0	43.9	49.2
May	42.1	54.8	50.1	21.1	17.0	17.6	46.7	53.9	51.5
June	0.7	26.0	32.8	26.8	24.6	22.3	34.4	47.6	45.9
July	32.4	0.2	14.9	27.4	27.7	26.2	46.0	37.6	43.2
August	8.3	0.4	9.8	27.4	27.9	25.7	40.6	39.7	44.5
T./Ave.	148.3	132.0	167.3	21.3	19.7	18.7	44.5	46.3	47.4
LAG: Long-Year Average (1978–2020), T: Total, Ave: Average

In order to determine whether the nutrients required by the plants were sufficient in the area where the experiment was established, soil samples were taken and mixed to represent the experimental area before planting and then analysed at Igdir University, Research Laboratory Application Centre (Table 3).

Table 3

Soil properties of the experimental area
Saturation (%)	pH	Total Salt (%)	Lime (%)	Organic Matter (%)	Total Nitrogen (%)	Phosphorus (P₂O₅) kg/da	Potassium (K₂O) kg/da
69.86	8.38	0.26	1.81	1.42	0.071	4.60	28.14
Clay-Loamy	Slightly Alkaline	Slightly Saline	Calcareous	Less	Moderately	Less	Moderately

Determination of plant growth promoting properties of the bacterial strains in the study

Bacterial cultures grown on Nutrient Agar (NA) media for 24–48 hours were used in all tests conducted to determine the plant growth promoting properties of bacterial strains. Nitrogen fixing ability of bacterial strains was determined on N-Free Solid Malate-Sucrose mediumColony development in the petri dish was considered a positive result (Cattelan et al. 1999). Potassium solubilization of bacterial strains was determined using Aleksandrov medium. The lightening of color (formation of a transparent area) around the colonies growing in the medium was accepted as positive for its potassium solubilization feature (Meena et al. 2014). Phosphorus solubilization of bacterial strains was tested in National Botanical Research Institutes' Phosphate Growth Medium (NBRIP-BPB) liquid medium. The color change in the medium (color change from dark purple to light blue or becoming transparent) was considered as a positive result (Nautiyal 1999). Calcium solubilization of bacterial strains was determined using Yeast Extract Dextrose (YDC) medium. The formation of a transparent zone around the bacterial colonies was considered as a positive result (Meena et al. 2014). The capacity of bacterial strains to produce 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase enzyme was tested in Dworkin and Foster (DF) Salt medium according to the procedure developed by Penrose and Glick (2003). Colony growth on the medium was considered ACC-deaminase positive. Bacteria were tested for siderophore production using Crom Azurol S (CAS) agar medium. The formation of orange colored areas around the colonies growing in the petri dish was evaluated as positive siderophore production of the strains (Louden et al. 2011).

Determination of the growth of bacterial strains at different pH and salt concentrations

Each bacterial strain tested was inoculated on NA medium with pH adjusted to 8, 8.5, 9, 9.5 and 10 and incubated at 27 ℃ for 48 hours and the bacteria that grew after incubation were recorded as positive. Since the soil of the experimental area was slightly saline, the growth of the tested bacterial strains was tested in media containing 1%, 3%, 5% and 7% sodium chloride (NaCl). For this purpose, the strains planted in NA medium were incubated at 27 ℃ for 48 hours. At the end of the time, the strains that showed growth were evaluated as positive and the strains that did not show growth were evaluated as negative (Olur 2020).

Determination of phytohormone (IAA, GA3, ABA, Zeatin and Salicylic Acid) production of bacterial strains in the study

The hormone production of the bacterial strains tested for their effect on plant growth was determined qualitatively and quantitatively using HPLC (High liquid pressure chromatography) device. Hormone extraction from bacterial strains was performed according to the method reported by Atıcı et al. (2003). The amount of hormones produced by the analyzed bacteria was calculated using the calibration graph method (y = 107.4x-58.5 for zeatin, y = 8.8x + 5.5 for gibberellic acid, y = 44.7x + 16.5 for indoleacetic acid, y = 245.4x-65.8 for abscisic acid and y = 48.2x-160.5 for salicylic acid). The calibration graphs of the hormones are given in Fig. 3.

The experiment was established in Iğdır University, Faculty of Agriculture, Agricultural Research and Application area (39°55'44"N 44°05'42"E), with three replications according to the Randomized Blocks Trial Design (Fig. 4) and was conducted in Iğdır, 2021 and 2022. It was carried out repeatedly for two years during the melon production season (March-August) and no product other than melon was grown in the trial area between the two periods. After the planting area was levelled by ploughing with a crowbar, it was parcellised with a distance of 70 cm above the row, 120 cm between the rows, 140 cm between the applications in the block and 200 cm between the blocks. There were 3 rows with 12 plants in each plot. Then, drip irrigation pipes were installed in the experiment area. There were 11 different applications (IT 22, IT 22 + fertilizer, IT 63, IT 63 + fertilizer, IT 93, IT 93 + fertilizer, IT 115, IT 115 + fertilizer, Mix, Mix + fertilizer and fertilizer only) (Table 4). During the period of the experiment, all cultural operations were carried out regularly.

Viols used for growing seedlings to be transplanted to the field were disinfected with 2% sodium hypochlorite and then placed in viols containing sterile soil and peat (1:1). Surface disinfected seeds were kept in bacterial solutions prepared at a density of 10⁶ cfu/ml for 2 hours and then dried. Then, one seed was sown in each cavity. The plants were transplanted to the field when they had 4–5 leaves (Fig. 5). The applications made after the seedlings were transferred to the field are given in Table 4.

Table 4

Applications made after the seedlings are transferred to the field
Applications	Information about applications
Bacteria application	When the plants were in the seedling stage, 5 ml of the solution of each bacterial strain was inoculated into the soil through the root collar of the plant and 100 ml/plant 1 month after the plants were transferred to the field.
Fertilizer application	Half of the fertilizer, adjusted as 13.45 kg da^− 1 N and 8.7 kg da^− 1 P in total, was applied to the plots before planting and half during the flowering period (Kokalis-Burelle et al. 2003)
Fertilizer + bacteria application	The plots in this application were treated with bacteria after the specified amount of fertilizer was applied.
Mix application	Solutions of 4 bacterial strains were taken in equal volumes and the mixture was prepared and applied to the soil through the root collar of the plant as indicated in the bacteria application.
Mix + fertilizer application	Fertilizer and mix applications were applied together

When the fruits reached harvest maturity, 3 plants representing the treated plots were selected and uprooted after removing the edge effect of the plots, and qualitative and quantitative measurements were made.

Yield and quality parameters examined in the study

The germination rates of the seeds (%) were determined according to Yıldırım and Güvenç (2006), and the main stem length (cm) was determined according to Kutsal (2017). While plant fresh weight (g) was determined according to Emrebaş (2010), chlorophyll content (%) in the leaf was measured according to Rostami et al (2008) using the SPAD-502 device. Shell thickness (mm), diameter (cm), length (cm) and flesh thickness (mm) of fruit, number of fruits per plant and fruit yield were determined according to the method of Karabulut (2018). Using the method reported by Eşiyok et al (2005), the average fruit weight and the amount of water-soluble dry matter (%) were measured, and the yield per decare (kg/da) was determined according to Kutsal (2017).

Statistical analysis

The significance levels of the variation sources of the data obtained in the study were determined in SPSS (17.0) statistical program according to the year replicated Coincidence Blocks Experimental Design and comparisons of the significant averages were made according to Duncan test. In addition, correlation analysis was performed to reveal the relationship between the parameters examined in the same statistical program.

The results of plant growth promoting properties of bacterial strains such as nitrogen fixation, potassium and phosphorus solubilisation, ACC-deaminase activity, phytohormone and siderophore production are given in Table 5. All of the tested bacterial strains were found to fix nitrogen, solubilise phosphorus and potassium and produce siderophore. The highest zeatin (51.8 ng/µl) and siderophore production (23.0 mm) was detected in Acinetobacter calcoaceticus strain IT 93 (Table 5). It was determined that all of the bacterial strains in the study showed growth in all of the media prepared at different pH and salt concentrations tested.

Table 5

Plant growth promoting properties of bacterial strains
Strains	N	K	P	Ca	ACC	S	Phytohormones(ng/µl)
Strains	N	K	P	Ca	ACC	S	GA	Z	ABA	IAA	SA
IT 22	K+	K+	K+	-	K+	21.0	-	16.5	-	-	-
IT 63	+	K+	K+	-	K+	14.0	14.6	-	-	-	-
IT 93	K+	K+	K+	-	K+	23.0	13.3	51.8	-	-	-
IT 115	K+	Z+	K+	-	K+	14.0	-	16.7	-	-	-
N: Nitrogen, K: Potassium, P: Phosphorus, Ca: Calcium, ACC: 1-aminocyclopropane-1-carboxylic acid-deaminase, S: Siderophore, GA: Giberallic acid, Z: Zeatin, IAA: Indole acetic acid, ABA: Abscisic acid, SA: Salicylic acid, Z+: Weak positive, K+: Strongly positive, +: Positive, -: Negative (IT 22: Bacillus safensis, IT 63: Acinetobacter calcoaceticus, IT 93: Acinetobacter calcoaceticus, IT 115: Serratia rubidaea)

Effect of bacterial strains on yield and quality parameters of melon plant

It was determined that 100% germination was realized in all seeds inoculated with bacterial strains. The results of the analysis of the effects of applications on main stem length, plant fresh weight, fruit diameter are presented in Table 6. According to the results of the statistical analysis, it was observed that the effect of year and bacteria application on all parameters examined was significant, while the bacteria application*year interaction was insignificant.

Table 6

Effect of bacterial applications on main stem length, plant fresh weight and fruit diameter
Applications	Main Stem Length (cm)			Plant fresh weight (g)			Fruit diameter (cm)
Applications	2021	2022	Mean	2021	2022	Mean	2021	2022	Mean
IT115	226.6	239.5	233.0^d**	1113.2	1216.6	1164.9^e**	16.7	20.1	18.4a-^c**
IT115 + Fertilizer	244.4	250.7	247.5^b − d	1305.6	1385.9	1345.7^cd	17.7	20.1	18.9^ab
IT22	234.5	243.3	238.9^d	1238.8	1331.7	1285.2^d	16.6	17.5	17.0^cd
IT22 + Fertilizer	252.4	263.5	258.0^a − d	1409.2	1464.3	1436.7^bc	17.7	19.7	18.7^a − c
IT63	238.8	253.3	246.0^b − d	1323.4	1379.1	1351.3^cd	16.4	17.5	17.0^cd
IT63 + Fertilizer	255.8	261.7	258.8^a − d	1385.4	1459.9	1422.6^bc	17.5	18.4	18.0^bc
IT93	247.0	263.6	255.3^b − d	1416.3	1483.1	1449.7^b	16.9	18.2	17.6^bc
IT93 + Fertilizer	264.0	274.2	269.1^a − c	1506.7	1626.4	1566.5^a	17.7	19.2	18.5^a − c
Mix	267.0	277.0	272.0^ab	1429.1	1493.9	1461.5^b	16.2	18.4	17.3^b − d
Mix + Fertilizer	281.1	288.5	284.8^a	1534.2	1686.5	1610.3^a	19.7	20.2	19.9^a
Fertilizer	232.3	249.0	240.7^cd	1249.9	1344.8	1297.3d	15.4	16.1	15.7^d
Year average	249.4^b	260.4^a*		1355.6	1442.9^**		17.1^b	18.7^a**
** %1, * %5. Data are averages of three replicates. The difference between values with the same letters in the same column is statistically insignificant (IT 22: Bacillus safensis, IT 63: Acinetobacter calcoaceticus, IT 93: Acinetobacter calcoaceticus, IT 115: Serratia rubidaea)

When the average values in Table 6 are examined, the highest main stem length (284.8 cm) in terms of applications was measured in the plants with Mix + Fertilizer application and followed by the Mix (272.0 cm) application. The lowest main stem length was measured as 233.0 cm and 238.9 cm, respectively, in IT 115 and IT 22 applications, which are in the same statistical group. When evaluated in terms of years, it was determined that the plants in 2022 had a higher main stem length compared to 2021. When the average values of plant fresh weight were examined, the highest values were obtained as 1610.3 g and 1566.5 g as a result of Mix + fertilizer and IT 93 + fertilizer applications, respectively. On the other hand, it was observed that the lowest fresh plant weight was obtained from IT 115 application with 1164.9 g. When evaluated in terms of years, it was determined that the fresh plant weight in 2022 (1442.9 g) was higher than in 2021 (1355.6 g). When the applications were evaluated in terms of fruit diameter, the highest fruit diameter values were measured in the Mix + Fertilizer application (19.9 cm) and the lowest (15.7 cm) in the control group.

The values of fruit length, fruit flesh and fruit shell thickness obtained as a result of the applications in the study carried out for two years are given in Table 7.

Table 7

Fruit length, fruit flesh and fruit shell thickness values obtained as a result of the applications
Uygulamalar	Fruit length (mm)			Flesh thickness (mm)			Shell thickness (mm)
Uygulamalar	2021	2022	Mean	2021	2022	Mean	2021	2022	Mean
IT115	21.8	22.8	22.3^bc*	33.5	35.0	34.3b^c**	11.7	10.4	11.0b^c**
IT115 + Fertilizer	23.6	25.0	24.3^ab	35.7	37.3	36.5^b	10.5	10.0	10.3^cd
IT22	21.4	22.8	22.1^bc	33.6	36.0	34.8^bc	12.5	12.0	12.3^a
IT22 + Fertilizer	23.2	24.3	23.7^a − c	36.0	37.0	36.5^b	11.5	11.3	11.4^ab
IT63	21.6	22.5	22.0^bc	33.2	33.9	33.5^c	13.3	11.5	12.4^a
IT 63 + Fertilizer	23.6	25.1	24.3^ab	34.3	36.1	35.2^bc	10.3	9.7	10.0^cd
IT93	21.9	22.7	22.3^bc	34.4	36.0	35.2^bc	12.4	11.4	11.9^ab
IT 93 + Fertilizer	22.3	23.2	22.8^a − c	34.3	36.9	35.6^bc	12.0	10.9	11.4^ab
Mix	22.6	24.4	23.5^a − c	35.1	37.4	36.3^b	11.2	10.6	10.9^bc
Mix + Fertilizer	23.8	25.5	24.7^a	37.7	39.6	38.7^a	9.7	9.1	9.4^d
Fertilizer	20.9	22.3	21.6^d	33.3	34.1	33.7^c	11.3	10.7	11.0^bc
Year average	22.4^b	23.7^a**		34.7^b	36.3^a**		11.5^a	10.7^b**
** %1, * %5. Data are averages of three replicates. The difference between values with the same letters in the same column is statistically insignificant. (IT 22: Bacillus safensis, IT 63: Acinetobacter calcoaceticus, IT 93: Acinetobacter calcoaceticus, IT 115: Serratia rubidaea)

When Table 7 was analysed, it was seen that Mix + fertiliser treatment had the highest fruit length and the control group had the lowest fruit length. When the results were evaluated in terms of years, it was determined that the fruit length was higher in 2022 compared to 2021. The highest fruit flesh thickness was 38,7 mm in the fruits of Mix + Fertilizer treatment, while the lowest flesh thickness was determined in the fruits of IT 63 and control applications and these two applications were statistically in the same group. On the other hand, fruit flesh thicknesses showed significant differences over the years, and the highest fruit flesh thickness was determined in 2022 with 36.3 mm. Fruit shell thicknesses showed significant differences depending on bacterial applications, and the best shell thickness of 9.4 mm was obtained as a result of Mix + Fertilizer application. The thickest fruit peel was measured in fruits in the IT 63 application, with 12.4 mm. (Table 7). When evaluated in terms of years, thinner fruit shell thickness was measured in 2022.

The analysis results of the effects of the applications on the amount of water-soluble dry matter, the number of fruits per plant and chlorophyll content in the leaf are given in Table 8.

Table 8

Water-soluble dry matter, leaf chlorophyll content and the number of fruits per plant obtained as a result of the applications
Applications	WSDM (%)			LCC (%)			NFPP (number/plant)
Applications	2021	2022	Mean	2021	2022	Mean	2021	2022	Mean
IT115	8.1	9.1	8.6^{b − d**}	67.6	70.6	69.1^ef**	2.21	2.46	2.34^d**
IT115 + Fertilizer	8.5	9.5	9.0^bc	70.5	74.5	72.5^d − f	2.27	2.58	2.43cd
IT22	8.0	9.2	8.6^b − d	68.5	73.3	70.9^d − f	2.20	2.42	2.31^d
IT22 + Fertilizer	8.8	9.8	9.3^ab	72.4	79.5	76.0^c − e	2.50	2.67	2.59^b
IT63	7.6	8.5	8.0^de	77.3	83.4	80.3^b − d	2.51	2.62	2.57^b
IT 63 + Fertilizer	8.3	9.6	9.0^bc	79.8	84.6	82.2^bc	2.57	2.71	2.64^ab
IT93	8.5	10.0	9.3^ab	75.8	83.1	79.5^b − d	2.56	2.69	2.62^ab
IT 93 + Fertilizer	8.9	11.0	9.9^a	83.7	88.4	86.1^b	2.60	2.73	2.67^ab
Mix	7.8	8.7	8.3^c − e	87.5	90.4	88.9^ab	2.63	2.76	2.70^ab
Mix + Fertilizer	8.3	9.3	8.8^b − d	91.6	102.0	96.8^a	2.69	2.84	2.77^a
Fertilizer	7.4	7.7	7.6^e	63.3	65.2	64.3^f	2.53	2.56	2.55^bc
Year average	8.2^b	9.3^a**		76.2^b	81.4^a**		2.48^b	2.64^a*
** %1, * %5. Data are averages of three replicates. The difference between values with the same letters in the same column is statistically insignificant. WSDM: Water-soluble dry matter, LCC: Leaf chlorophyll content, NFPP: Number of fruits per plant (IT 22: Bacillus safensis, IT 63: Acinetobacter calcoaceticus, IT 93: Acinetobacter calcoaceticus, IT 115: Serratia rubidaea)

In terms of bacterial applications, the amount of water-soluble dry matter varied between 7.6% and 9.9%. The highest and lowest amounts of water-soluble dry matter were obtained as a result of Acinetobacter calcoaceticus 93 + Fertilizer and control applications, respectively. When the amount of water-soluble dry matter was evaluated by years, it was seen that a higher amount of water-soluble dry matter was obtained in 2022. Chlorophyll content in the leaf showed a significant difference depending on the bacterial applications and the highest chlorophyll content was 96.8% in Mix + fertilizer treated plants. In terms of bacterial applications, when the number of fruits per plant was considered, the highest number of fruits (2.77) was determined in the Mix + Fertilizer application. It was observed that the number of fruits per plant was higher in 2022 (Table 8).

The average fruit weight, per plant and per decare fruit yield values obtained as a result of the practices and the averages of these values are presented in Table 9.

Table 9

Effect of bacterial applications on average fruit weight and melon yield
Applications	AFW (g/fruit)			FYPP (g/plant)			FYPD (kg/da)
Applications	2021	2022	Mean	2021	2022	Mean	2021	2022	Mean
IT115	2353.0	2569.9	2461.5^e**	5200.4	6313.5	5757.0^h**	4368.4	5303.3	4835.8^h**
IT115 + Fertilizer	2649.1	2806.0	2727.5^cd	6017.5	7239.1	6628.3^fg	5054.7	6080.9	5567.8^fg
IT22	2623.2	2538.5^de	2623.2	5399.2	6351.5	5875.3h	4535.3	5335.2	4935.3^h
IT22 + Fertilizer	3003.3	3131.7	3067.5^b	7492.0	8357.5	7924.7^bc	6293.3	7020.3	6656.8^bc
IT63	2386.3	2478.3	2432.3^e	5969.6	6482.7	6226.1^gh	5014.5	5445.4	5230.0^gh
IT 63 + Fertilizer	2656.6	2842.0	2749.3^cd	6824.9	7699.9	7262.4^de	5733.0	6467.9	6100.4^de
IT93	2876.2	2985.4	2930.8^bc	7359.2	8017.1	7688.1^cd	6181.7	6734.3	6458.0^cd
IT 93 + Fertilizer	2944.8	3092.7	3018.8^b	7653.2	8446.8	8050.0^bc	6428.7	7095.3	6762.0^bc
Mix	3014.0	3134.3	3074.2^b	7936.4	8654.0	8295.2^b	6666.6	7269.3	6968.0^b
Mix + Fertilizer	3234.1	3380.8	3307.5^a	8683.9	9608.6	9146.2^a	7294.4	8071.3	7682.8^a
Fertilizer	2660.0	2801.6	2730.8^cd	6740.6	7174.7	6957.6^ef	5662.1	6026.7	5844.4^ef
Year average	2748.3^b	2895.1^a**		6843.3^b	7667.8^a**		5748.4^b	6440.9^a**
** %1, * %5. Data are averages of three replicates. The difference between values with the same letters in the same column is statistically insignificant. AFW; Average fruit weight, FYPP; Fruit yield per plant, FYPD; Fruit yield per decare (IT 22: Bacillus safensis, IT 63: Acinetobacter calcoaceticus, IT 93: Acinetobacter calcoaceticus, IT 115: Serratia rubidaea)

Considering Table, it is seen that the average fruit weight varies between 2432.3 g and 3307.5 g, depending on bacterial applications. When evaluated in terms of years, a higher average fruit weight was achieved in 2022 compared to 2021. The highest fruit yield per plant (9146.2 g/plant) and fruit yield per decare (7682.8 g/plant) were determined in the Mix + Fertilizer application (Fig. 6). After this application, the highest value was determined in the Mix application, where strains were used as a mixture.

In order to test whether there is a relationship between the yield parameters examined in the current study and to reveal the degree of the existing relationship, the correlation coefficients of the examined parameters were calculated and their significance levels are presented in Table 10.

Table 10

Correlation coefficients and significance levels of yield and quality parameters
	AFW	NFPP	FYPP	FYPD	FT	FL	ST	FD	WSDM	LCC	PFW	MSL
AFW	1.00	0.23^n.s.	0.90^**	0.90^**	0.50^**	0.36^**	− .22^n.s.	0.38^**	0.48^**	0.29^*	0.52^**	0.35^**
NFPP		1.00	0.62^**	0.62^**	0.24^*	0.23^n.s.	− .15^n.s.	0.24^n.s.	0.24^n.s.	0.42^**	0.32^**	0.25^*
FYPP			1.00	1.00^**	0.52^**	0.39^**	-0.24^*	0.42^**	0.49^**	0.43^**	0.56^**	0.39^**
FYPD				1.00	0.52^**	0.39^**	-0.24^*	0.42^**	0.49^**	0.43^**	0.56^**	0.39^**
FT					1.00	0.38^**	− .20^n.s.	0.39^**	0.17^n.s.	0.24^n.s.	0.50^**	0.22^n.s.
FL						1.00	-0.44^**	0.35^**	0.15^n.s.	0.50^**	0.41^**	0.54^**
ST							1.00	-0.35^**	-0.08^n.s.	-0.19^n.s.	-0.15^n.s.	− .15^n.s.
FD								1.00	0.38^**	0.52^**	0.57^**	0.14^n.s.
WSDM									1.00	0.22^n.s.	0.49^**	0.18^n.s.
LCC										1.00	0.67^**	0.42^**
PFW											1.00	0.34^**
MSL												1.00

** %1, * %5, n.s.: non-significant. FYPP; Fruit yield per plant, AFW; Average fruit weight, NFPP: Number of fruits per plant, FYPD; Fruit yield per decare, FT: Flesh thickness, FL: Fruit length, ST: Shell thickness, FD: Fruit diameter, WSDM: Water-soluble dry matter, LCC: Leaf chlorophyll content, PFW: Plant fresh weight, MSL: Main Stem Length.

Table 10 shows that there is a very significant and positive relationship between average fruit weight and fruit yield per plant and per decare, and a very significant and moderately positive relationship between fruit flesh thickness, fruit length, fruit diameter, water soluble dry matter, plant fresh weight and main stem length. There was a very significant and positive relationship between the number of fruits per plant and fruit yield per plant and fruit yield per decare, and a very significant but moderately positive relationship between the number of fruits per plant and leaf chlorophyll content and plant fresh weight. While a very significant and quite high relationship was observed between fruit yield per plant and fruit yield per decare, a very significant and moderately positive relationship was found between fruit yield per plant and fruit flesh thickness, fruit length, fruit diameter, water soluble dry matter content, leaf chlorophyll content, plant fresh weight and main stem length. There was a very significant and moderately positive relationship between fruit yield per decare and fruit flesh thickness, fruit length, fruit diameter, water soluble dry matter content, leaf chlorophyll content, plant fresh weight and main stem length. It was observed that there was a very significant and moderately positive relationship between fruit flesh thickness and fruit length, fruit diameter and plant fresh weight. There was a very significant and moderately inverse relationship between fruit length and fruit skin thickness and a very significant and moderately positive relationship between fruit length and fruit diameter, leaf chlorophyll content, plant fresh weight and main stem length. There was a very significant and moderately negative relationship between fruit length and fruit skin thickness. A very significant and moderately negative relationship was found between fruit skin thickness and fruit diameter. There was a very significant but moderately positive relationship between fruit diameter and water soluble dry matter content, leaf chlorophyll content and plant fresh weight. A very significant and moderately positive relationship was found between water soluble dry matter content and plant fresh weight. There was a very significant and moderately positive relationship between leaf chlorophyll content and plant fresh weight and main stem length. There was a very significant and moderate positive relationship between plant fresh weight and main stem length.

The use of bacterial strains in the nutrient cycle as biofertilizers provides a very important advantage for minimizing the negative effects caused by commercial fertilizers in agricultural production and for sustainable agriculture (Öztekin et al. 2015). For this reason, in the current study, the effect of bacterial strains on melon cultivation under field conditions was investigated and it was found that Mix + Fertilizer application increased fruit yield per decare by 32% compared to the control group with only fertilizer application. This result showed that bacterial strains also increased the effectiveness of the applied fertilizer. The most desired thing in agricultural production is to increase productivity by making maximum use of the applied fertilizer. In different studies, it has been determined that bacterial strains increase plant growth by producing siderophores and phytohormones, fixing nitrogen, solubilizing phosphorus and potassium, and synthesizing stress-relieving enzymes. Additionally, it has been determined that some bacteria improve root development by increasing the accessibility of essential nutrients and support plant development by reducing the damage caused by stress by activating plant defense systems (Franche et al. 2009; Alves et al. 2015). Studies have shown that nitrogen, one of the most vital elements in plant growth and various metabolic activities, is converted by both symbiotic and free-living microorganisms into a form that plants can use by the nitrogenase complex (Dixon and Kahn 2004; Ali et al. 2017), and that the structural and nitrogenase complex consists of regulatory genes involved in the biosynthesis of Fe protein and Fe-molybdenum cofactor activation (Parray et al. 2016). Additionally, some nitrogen-fixing rhizobacteria species have been found to have the ability to attenuate the increase in ethylene levels by synthesizing rhizobiotoxin, which inhibits the function of the ACC synthase enzyme (Vejan et al. 2016; Gouda et al. 2018). The mineralisation of phosphorus, which has important roles in various plant metabolic processes such as photosynthesis and energy transfer (Sharma et al. 201), by microorganisms has been found to increase the availability of phosphorus in soil and mineralisation is highly dependent on soil pH. Such microorganisms have been found to alter soil pH by secreting various organic and inorganic acids (malic, malonic, fumaric, oxalic, acetic, tartaric, glutanic, propoinic, butyric, lactic, 2-keto gluconic, gluconic and glyconic acids) through a mechanism known as rhizosphere acidification (Kumar et al. 2018; Umar et al. 2020). It is known that potassium, which plays a key role in the processes related to growth and development in plants, plays a role in activating more than 80 enzymes involved in various energy metabolic processes (Adams and Shin 2014; Wang et al. 2017). Potassium-solubilizing bacteria have been found to produce low molecular weight organic acids, including lactic, tartaric, glycolic, fumaric, oxalic, citric, malic, gluconic and 2-ketogluconic acid, and to release potassium from fixed potassium-containing minerals (Liu et al., 2012; Prajapati et al., 2013; Saiyad et al., 2015). It has been stated that presence of iron in plants, which is essential for the survival of all organisms, is associated with the production of low molecular weight siderophores produced by bacteria. It has been determined that siderophores show high affinity for Fe3+, reduce the population of pathogenic microorganisms and increase plant growth by creating iron competition in the rhizospheric region (Saha et al. 2016; Hakim et al. 2021). Considering that all of the strains tested in this study have the ability to fix nitrogen, dissolve phosphorus and potassium, produce siderophores and change soil pH with the acids they secrete, it can be stated that these properties play a role in increasing the development of the melon plant. In addition, bacterial strains have been found to help modulation of hormone concentrations such as auxins (indoleacetic acid) and cytokinins in plants. Another important mechanism by which plant growth is promoted through bacterial strains is the reduction of plant ethylene levels, a hormone produced by plants under stress conditions (Gamalero and Glick 2015). It has been shown that ACC (1-aminocyclopropane-1-carboxylate) produced by PGPB is hydrolyzed by the 1-aminocyclopropane-1-carboxylate deaminase enzyme as a direct precursor of ethylene hormone in plants (Orozco-Mosqueda et al., 2020). The bacterial ACC deaminase enzyme has been found to function by degrading the ACC molecule into α-ketobutyrate and ammonia, thereby preventing the increase in ethylene production under various stress conditions, thus promoting the growth of plants and facilitating their survival (Santoyo et al. 2020). The fact that all four strains tested in this study gave positive results for ACC deaminase shows that they are important in supporting plant development. In the current study, it was determined that seed coding with bacteria had a positive effect on seed germination and increased the germination rate compared to the control. Johnson et al. (2011) reported that strains in the cytokinin group that produced zeatin and also gibberallic acid had a positive effect on germination in melon seeds. It has been reported that this is due to the fact that PGPBs promote the synthesis of growth regulatory substances such as gibberellins, which trigger the activity of germination-specific enzymes such as ɑ-amylase, proteases and nucleases. Johnson et al. (2011) reported that strains in the cytokinin group that produced zeatin and also gibberallic acid had a positive effect on germination in melon seeds. The same researchers reported that this is due to the fact that PGPBs promote the synthesis of growth regulatory substances such as gibberellins, which trigger the activity of germination-specific enzymes such as ɑ-amylase, proteases and nucleases. Furthermore, IAA produced by bacteria is known to provide an ecological advantage to bacteria by promoting environmental adaptation under stress conditions such as UV, salt and acidity (Bianco et al. 2006). In addition, it is suggested that IAA acts as a signaling molecule within microorganism cells and thus provides successful results by increasing the chance of survival in the microbiome (Matsukawa et al. 2007). In parallel with this result, when the current study findings were evaluated, zeatin production was detected in 3 of the 4 strains tested for their effect on yield in melon and GA production was detected in 2. This result was found to be compatible with the findings of Kumar et al (2014) and Vishwas et al (2017), which reported that PGPBs significantly affected germination. In a study conducted with Bacillus, Pseudomonas, Azotobacter and Acinetobacter species, it was found that bacterial applications increased the main stem length and number of fruits per plant in bitter melon compared to the control (Singh and Jha 2017). It has been reported that 9700 kg da-1 fruit yield was obtained as a result of root applications with bacterial strains in melon plants. It was also reported that the strains increased the shoot fresh weight (1186.2 g), root fresh weight (128.6 g) and main stem length (482.2 cm) of melon grown under greenhouse conditions compared to the control (Abraham Juárez et al. 2018). In the current study, it was determined that bacterial applications increased the number of fruits per plant compared to the control and the highest main stem length (284.8 cm) and fruit yield per decare (7682.8 kg) were obtained in the Mix + Fertilizer application. In different studies, it has been reported that applications with bacterial strains belonging to the Pseudomonas, Serratia and Bacillus genera significantly increased plant fresh weight, main stem length, number of fruits per plant, fruit diameter, average fruit weight and total fruit yield compared to the control (Almaghrabi et al. 2013; Yıldırım et al. 2022; Dönmez et al. 2024). In the present study, it was determined that the peel thickness varied between 9.4–12.4 mm whileIn another study in which the effect of bacterial applications on melon yield was tested, fruit yield of 2000–2300 kg/ha and fruit peel thickness values ranging between 13,58 − 14,56 mm were obtained (Kutsal 2017). It is seen that the melon variety, the difference in bacterial strains and the region (different climate and soil characteristics) are effective in the values of the obtained parameters. It was determined that all applications in the study increased the chlorophyll content in the leaf compared to the control and the highest chlorophyll content (96.8%) was obtained from the Mix + Fertilizer application. It was reported by Kaçar (2015) that there is a positive relationship between chlorophyll content and yield and that an increase in chlorophyll content increases plant yield. As stated, in the current study, the highest yield per decare was obtained from the Mix + Fertilizer application, where the highest chlorophyll content was measured, and the results are parallel in this sense.

In the current study, it was seen that Mix and Mix + Fertilizer applications stood out among the applications with the best results. It has been reported that the combined application of strains with different mechanisms of action may show higher activity compared to single application (Spaepen et al. 2007). It has been reported that the use of bacterial strains in combination provides effectiveness in different environmental conditions by using multiple mechanisms of action, overcomes inconsistencies caused by the use of strains individually, and increases the success of strains by stimulating the production of various enzymes and metabolites (Santhanam et al. 2019). This approach was found to be compatible with the results of the current study. In addition, the effect of bacteria on plant development; It has also been reported that it varies depending on the tested plant species, inoculation method and inoculum density (Mangmang et al. 2015). In addition to the genotype, which is primarily effective in increasing yield and quality, environmental factors such as temperature, light, humidity and CO2 also play an important role (Özkaraman 2004). The increase in photosynthesis leads to an increase in dry matter, and the increased dry matter accumulates in the stem and storage organs and causes an increase in stem diameter. Studies on this subject indicate that dry matter accumulation in plants varies according to temperature, technical and cultural processes and affects the distribution of the produced dry matter to the organs of the plant (Uzun et al. 1998; Sarıbaş et al. 2018). When the temperature values for the period in which the experiment was conducted in the current study were examined, higher temperatures were observed in 2021 compared to 2022, especially during the flowering period of the plants. It has been reported that an increase in temperature increases vegetative development, but causes both a decrease in the formation of female flowers and a later onset of flowering, which reduces generative development and thus leads to a decrease in yield (Özkaraman 2004). In this context, it was seen that the results of the current study were parallel and when the results obtained were evaluated, higher values were obtained in terms of the parameters examined, especially melon yield, in the second year of the study.

Recently, chemical fertilizers used unconsciously to increase productivity and quality in agricultural areas cause a decrease in soil fertility, environmental pollution and threaten the food security of the world population. In reducing the use of costly and highly polluting chemical fertilizers, applying bacterial strains as a mixture will help protect environmental health and maintain soil fertility. When the results are evaluated in terms of melon development, it can be seen that Mix + fertilizer and Mix applications have very successful effects on increasing yield. Higher fruit yield was obtained from IT 93 and Mix application without fertilizer use compared to the control. These results showed that bacterial applications can be used as an important alternative to reduce the use of chemical fertilizers. When bacterial strains were used together with fertilizer, the best values were obtained for the examined parameters. The application of such strains will enable sustainable use of natural resources and improvement of crop production by reducing the availability of available nutrients and the application of chemical fertilizers.

Ethical approval

Not applicable.

Consent to participate

Not applicable.

Consent for publication

All the authors approved the fnal version of the manuscript.

Competing interests

The authors declare no competing interests

Funding

We would like to thank the Iğdır University Department of Scientific Research Projects (Project No. ZİF0322D01) for the financial support in conducting this study.

Author contributions

IT and MFD co-conducted the study, wrote, read and approved the first draft of the article.

Data availability

Presented data is the part of PhD research work of Işıl Temel (IT) and is available from the corresponding author.

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The Role of Bacteria Isolated from Different Sources in Melon Cultivation: Effects on Yield and Quality

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Abstract

Figures

Introduction

Material and methods

Determination of plant growth promoting properties of the bacterial strains in the study

Determination of the growth of bacterial strains at different pH and salt concentrations

Yield and quality parameters examined in the study

Statistical analysis

Results

Effect of bacterial strains on yield and quality parameters of melon plant

Discussion

Conclusions

Declarations

Funding

Author contributions

Data availability

References

Supplementary Files

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