effect of mycorrhizae inoculation on plant growth, yield, and ...

COMMUN. SOIL SCI. PLANT ANAL., 33(13&14), 2189–2201 (2002)

EFFECT OF MYCORRHIZAE INOCULATION ON PLANT GROWTH, YIELD, AND PHOSPHORUS UPTAKE IN GARLIC UNDER FIELD CONDITIONS Nebahat Sari,1,* I˙brahim Ortas,2 and Halit Yetisir1 Departments of 1Horticulture and 2Soil Science, Faculty of Agriculture, University of C¸ukurova, Adana, Turkey

ABSTRACT The effect of mycorrhizae species and phosphorus (P) fertilizer on garlic (Allium sativum L.) growth, yield –quality, and P uptake under high P accumulated non-sterile field conditions were studied. Experiments were conducted for two successive years under field conditions of Menzilat soil series (Typic xerofluvent) at the Research Farm of the University of Cukurova (Turkey). Glomus mosseae arbuscular mycorrhizae (AM) were tested on local Urfa genotype of garlic at 0, 40, 80, and 120 kg phosphorus (P2O5) ha21. In the first year, garlic was inoculated with 1,000 spores per plant, but in the second year, garlic was inoculated with either 1,000 or 2,000 spores per plant. Emergence, plant growth, yield, bulb size, root mycorrhizal infection, and phosphorus uptake of plants were examined. Neither mycorrhizal inoculation nor P2O5 supply increased garlic growth and yield. However, at 0 level of P2O5 application, mycorrhizal inoculation slightly increased plant P uptake. In the *Corresponding author. E-mail: [email protected] 2189 Copyright q 2002 by Marcel Dekker, Inc.

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SARI, ORTAS, AND YETISIR

second year of the experiment, mycorrhizae significantly increased clove yield. The results revealed that although garlic is mycorrhizal dependent, mycorrhizal inoculation did not contribute to the plant growth and nutrient uptake. Key Words: Garlic; G. mosseae; Phosphorus uptake; Soil; VA mycorrhizae; Yield

INTRODUCTION About 500,000 ha of garlic (Allium sativum L.) are cultivated worldwide and total world garlic production is about 12 million metric tons. In Turkey, total garlic production is about 106,000 tons.[1] Garlic, believed to be a powerful antiseptic, contains 61.3% water and 38.7% dry matter of each 100 g of garlic. There are 6.2 g protein, 0.2 g oil, 30.8 g carbohydrate, 29 mg calcium (Ca), 1.5 mg iron (Fe), 202 mg P, 529 mg potassium (K), 19 mg sodium (Na), and some amount of vitamins B and C.[2] Garlic is an important and useful vegetable for human health because it has antibiotic properties, stimulates the heart, cleans the blood, regulates blood circulation, balances hypertension and helps asthma and bronchial problems.[3] Healthy plants can only grow in healthy environments. Plants take up nutrients easily and productively in soil where the biological balance is not broken. One of the recognized side effects of soil disinfections is the destruction of arbuscular mycorrhizal (AM).[4] Soil disinfection causes reduction in yield and quality, because plants do not grow well in soil where biological life has been devastated.[5] Allium spp., including garlic, are responsive to AM symbiosis. In the absence of AM fungi, onion growth is stunted.[6,7] Since the size of harvested garlic bulbs are directly related to the planted clove size, stunting in propagation material reduces not only the yield of the crop of propagation material, but also the yield of the subsequent commercial crop.[8] AM fungi application to soil or propagation material has been reported to be effective in suppressing this stunting in some other crops after soil disinfection.[9] AM living symbiotically on plant roots facilitate plant nutrient uptake by plant. The addition of AM to leek (Allium porrum ) was equivalent to the addition of 250 kg P2O5 ha21.[9] For the last 20 years, it has been frequently reported that in some soil there is a high P accumulation after a high amount of P fertilizer application. Accumulated P has a negative effect on the environment and soil quality parameters. Especially under the irrigation systems, the environment is polluted by excessive P. It is environmentally sound to utilize soil excess P by using some biological sources such as mycorrhiza and plant root mechanisms for plant growth and P uptake under such condition.

MYCORRHIZAE INOCULATION AND GARLIC

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In this study, G. mosseae mycorrhizae species with three different P2O5 levels was applied to undisinfected open fields for 2 successive years. The objective of this experiment was to study the effects of mycorrhizae inoculation on yield and quality of garlic under high P accumulated non-sterile field condition.

MATERIALS AND METHODS The study was established on Menzilat soil series,[10] located on the Research Farm of the University of Cukurova, Turkey, in 1997 – 1998 and 1998 – 1999. Some properties of the soil series are given on Table 1.

Plant and Mycorrhizal Material Turkish local garlic cultivar “Urfa local” was used as plant material. Inoculum of Glomus mosseae[11] (isolated from Rothamsted, UK), 1,000 spores/plant mix of source (soil, sand, and organic matter mix), chopped roots and mycorrhiza spores were placed 50 mm below the cloves.

Experimental Layout Experiments were set up for two succession years. In the first year, the soil was amended with 0, 40, 80, and 120 kg P2O5 ha21 and G. mosseae species. Mycorrhizae were applied to half of each plot (M þ ), but not to the other half (M 2 ). In the mycorrhizal plot, 1,000 spores/plant were applied under each clove. In the second year, two mycorrhizae doses were tried: 1,000 spores per plant (M1) and 2,000 spores per plant (M2). No fertilization was done in the first year except P2O5; but in second year, 160 kg ha21 N (as NH4NO3) and K2O (as K2SO4) were applied twice by dividing in two equal amounts during vegetation. The experiment was designed as a split block, with the main plots as P2O5 doses, and the mycorrhizal doses as the subplots. In both years, cloves were planted in each plot with 30 £ 10 cm spacing. Each subplot was replicated three times. Emergence plant height (cm) and stem diameter (cm) were recorded in each plot, once in the first year (31.3 in 1998), and twice in the second year (5.02 and 5.03 in 1999). Ten plants from each plot were randomly selected and the level of mycorrhizal infection on roots and shoot P concentration were determined. Stems of plants were broken to increase dry matter accumulation and, after one week, bulbs were harvested. Harvested bulbs were dried for 1 week in the greenhouse and yield with stem were determined for each plot. Bulb weight, bulb

b

a

7.85 7.81

pH (2:5 H2O) 0.172 0.116

N (%) 278 293

P2O5a (kg/ha) 900 800

Kb (ppm) 0.048 0.050

Salt (%) 30.4 29.3

CEC (meq/100 g)

Chemical and Biological Characteristics of Menzilat Soil Series

0.5 N NaHCO3 extraction (Olsen methods). K extracted by boiling with 1 N HNO3.

0– 20 20– 40

Soil Depth (cm)

Table 1.

72 39

Number of Mycorrhizal Spore (10 g Soil)

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diameter, and number of cloves in bulb were determined for 10 bulbs. In the second year, bulb height was also added to these parameters. AM spore counting was accessed according to wet the sieving method.[11] Roots were cleaned and stained according to Koske and Gemma.[12] Stained roots were observed under a stereomicroscope at a magnification of 40 £ by the method of Giovannetti and Mosse.[13] Phosphorus concentration was determined according to Murphy and Riley.[14] Results were evaluated on COSTAT statistical program with Tukey’s test by comparing means.

RESULTS First Year Results In the first year, plants started to emerge 10 –13 days after planting. Neither P doses nor mycorrhizal inoculation affected emerging time. Plant height and stem diameter measurement done on March 31, 1997 (Table 2). Neither P nor mycorrhizae affected plant height and stem diameter. In every treatment, plant height and stem diameter were between 93– 100 cm and 13– 14 mm, respectively. At the beginning, with mycorrhizal inoculation, color, appearance, and vigor of treated plants were better than untreated plants, but this difference among treatments disappeared. The highest yield (average 13.772 tons ha21) was determined in the 80 kg P2O5 ha21 treatment, but differences among treatments were not significant. Although mycorrhizal inoculation had no statistical effect on yield, inoculation slightly increased yield in low level of P treatment. These increases were 5%, 12%, and 4% in 0, 40, and 80 kg P2O5 ha21 treatments, respectively. Regardless of phosphorus application, the mean yield of all P treatments was

Table 2.

The First Year Plant Height (cm) and Stem Diameter (mm) Plant Length (cm)

Stem Diameter (mm)

P2O5 (kg/ha)

M2

Mþ

M2

Mþ

0 40 80 120 Mean D 5% (mycorrhizae) D 5% (P)

101.20 98.40 101.20 96.20 99.25

92.90 99.30 100.00 93.10 96.33

14.10 13.00 14.20 13.60 13.73

13.80 14.30 12.60 13.00 13.43

n.s. n.s.

n.s. n.s.

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13.001 tons ha21 in the inoculated plots and 12.380 tons ha21 yield in the noninoculated plots (Table 3). In harvested bulbs, neither P levels nor mycorrhizal inoculation affected bulb size or number of cloves per bulbs. Mean bulb weight, diameter, and number of cloves per bulb were 32.22 g, 43.09 mm, and 33.22 numbers, respectively (Table 3). In the first year experiment, the result of root infection and P content of leaf belongs to the measurement which was done at the middle of the vegetation period (Table 4). Mycorrhizal inoculation rate was higher in mycorrhizae amended plots than non-amended plots. The percentage colonization was 63.54% in AM inoculated plots and 52.53% in uninoculated. Plants in inoculated plots absorbed more P than non-inoculated plots. Plants inoculated with AM in 0 kg P2O5 ha21 dose had 16% more P than noninoculated plots. In 40 kg P2O5 ha21 dose, plants inoculated with AM had 42% more P than non-inoculated plants. Plants in 40, 80, and 120 kg P2O5 ha21 applied took place in the same group (a). Plots that P2O5 was not applied were located in another group (b). No statistically importance was found in P uptake of 80 and 120 kg P2O5 ha21 applications.

Second Year Results Plant emergence was unaffected by mycorrhizal application in the second year of the experiment (about 10– 12 days after planting in each plot). There was no interaction between mycorrhizae and P doses for any parameter. Plant length and stem diameter were unaffected significantly by applications (Tables 5 and 6). Plant length in plot without mycorrhizae was 57.96 cm, 57.55 cm in low-level mycorrhizae application and 56.88 cm in high-level mycorrhizae application. In the second measurements done one month later, the mean plant lengths were 95.89, 96.83, and 96.0 cm in M 2 , M1, and M2, respectively. Stem diameter measurements during February were 12.20 mm, 12.06 mm, and 12.01 mm in M 2 , M1, and M2 treatments, respectively. Similar relative results were obtained in March. No statistically difference was found in either measurement. No interaction was observed for yield and bulb characteristics or significant differences in inoculation (Table 7). The highest yield was obtained from M2 mycorrhizae application. Yields were 15.056, 15.029, and 15.657 tons ha21 in M 2 , M1, and M2 plots, respectively. On the other side, P doses did not affect yield. Nevertheless, the highest yields were taken from 80 –120 kg ha21 with yields of 15.615 and 15.614 tons ha21. The lowest yield was obtained from 40 kg P2O5 ha21 with 14.194 tons ha21. Bulb size and number of clove in bulb were unaffected by P applications, but number of cloves was affected by mycorrhizae inoculation and the highest number of clove was determined in both mycorrhizae dose inoculation.

M2 12.212 11.009 13.477 12.821 12.380

P2O5 (kg/ha)

0 40 80 120 Mean D 5% (mycorrhizae) D 5% (P) n.s. n.s.

12.863 12.321 14.066 12.755 13.001

Mþ

Yield (ton/ha)

32.44 29.73 31.34 33.36 31.72

M2

n.s. n.s.

32.28 30.14 31.26 37.19 32.72

Mþ

Bulb Weight (g)

44.41 41.79 40.63 44.12 42.74

M2

n.s. n.s.

44.21 41.88 43.07 44.56 43.43

Mþ

Bulb Diameter (mm)

Table 3. Yield and Bulb Characteristics of the First Year Experiment

32.90 32.90 32.20 34.60 33.15

M2

n.s. n.s.

32.90 31.70 31.50 37.10 33.30

Mþ

Number of Clove (Number/Bulb)

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Table 4.

Rate of Fungal Inoculation and P Content of Leaf in the First Year Experiment Rate of Inoculation (%)

P Content (%)

P2O5 Doses (kg/ha)

M2

Mþ

M2

Mþ


40.27 54.67 59.41 55.77 52.53 b

59.83 61.00 68.68 64.67 63.54 a

0.25 0.31 0.37 0.37 0.37

0.29 0.44 0.38 0.36 0.33

Table 5.

8.22 n.s.

n.s. 0.06

Plant Length in Two Different Dates in the Second Year (mm) 05/02/1999

05/03/1999

P2O5 (kg/ha)

M2

M1

M2

M2

M1

M2


58.41 57.35 58.04 58.00 57.96

56.60 57.53 59.00 56.90 57.55 n.s. n.s.

58.73 54.20 58.65 56.00 56.88

96.73 95.18 95.60 96.01 95.89

95.14 95.13 97.90 97.20 96.83 n.s. n.s.

95.86 93.30 98.28 96.60 96.00

Table 6. (mm)

Results of Stem Diameter Measured in Two Different Dates in the Second Year 05/02/1999

05/03/1999

P2O5 (kg/ha)

M2

M1

M2

M2

M1

M2

0 40 80 120 Mean D 5% (Mycorrhizae) D 5% (P)

12.27 11.46 12.47 12.63 12.20

11.36 12.29 12.47 12.18 12.06 n.s. n.s.

11.85 11.37 12.89 11.70 12.01

18.06 18.09 18.00 19.00 18.32

18.59 17.36 17.86 18.86 18.25 n.s. n.s.

18.22 16.79 17.63 17.70 17.57

M2 16.760 13.187 15.773 14.503 15.056

P2O5 Doses (kg/ha)


14.547 13.573 15.780 16.217 15.029 n.s. n.s.

M1

Yield (ton/ha)

15.387 15.823 15.293 16.123 15.657

M2 47.00 46.80 43.20 49.20 46.55

M2 41.50 37.70 46.96 47.20 43.34 n.s. n.s.

M1 42.70 43.23 47.80 46.70 45.11

M2

Bulb Weight (g)

49.82 51.16 48.80 49.93 49.93

M2 48.70 47.50 50.50 50.70 49.35 n.s. n.s.

M1 48.20 49.10 51.50 49.54 49.59

M2

Bulb Diameter (mm)

35.00 35.87 33.90 35.60 35.09

M2

33.97 32.80 35.50 34.30 34.14 n.s. n.s.

M1

34.90 36.30 36.80 33.40 35.35

M2

Bulb Height (mm)

Table 7. Yield and Bulb Characteristics of the Second Year Experiment

29.50 30.96 32.90 35.26 32.16 b

M2

36.90 33.80 38.90 38.50 37.03 a 4.61 n.s.

M1

M2 34.30 38.70 41.90 39.50 38.60 a

Number of Clove (Number/Bulb)

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Mycorrhizal inoculation doses significantly affected root infection, whereas P application had no affect (Table 8). Inoculation rate increased with concentration of mycorrhizae applied. There was still a high percentage of root infection in non-inoculated plots. Increasing P application caused in high level of P concentration in leaf. The increases changed between 9% and 18%. The highest increment was determined in 40 kg ha21 P application. No further increase was determined with increasing P application. DISCUSSION Although garlic and other Liliaceae species are mycorrhizal,[14,19] AM and P2O5 addition to the soil had little or no effect on yield and quality parameters in Urfa local garlic cultivar for both experiments. Under our experimental conditions, AM application did not contribute to plant yield and P uptake compared to noninoculated plots. The lack of any effect of AM application may be attributed to three reasons. First and most important is the presence of native mycorrhizae population. Native mycorrhizae likely masked effect of applied mycorrhizae. Second, the selection of inoculated species of mycorrhizae may be wrong for garlic plants. Each plant species has different response to different mycorrhizae species. In earlier results (unpublished data), under the greenhouse conditions showed that G. etunicatum was the most effective species for cucumber. However, garlic needs to be tested for better mycorrhiza species selection. Koch et al.[4] compared fumigated plots with AM added to unfumigated plots and without AM added plots and found no statistically significant differences between the two treatments. In the first year of experiment, mycorrhizae and P mean yield was 12.691 tons ha21 and the second year mean yield was 15.247 tons ha21. The reason of increasing in yield was related to the additional of N and K2O fertilization. But in the first year, no additional organic Table 8. Rate of Fungal Inoculation and P Content of Leaf in the Second Year Experiment Inoculation Rate (%)

P (%)

P2O5 (kg/ha)

M2

M1

M2

M2

M1

M2


30.0 42.0 31.0 36.0 34.7 c

35.0 43.0 39.0 37.0 37.9 b 7.9 n.s.

37.0 43.0 52.0 43.0 43.7 a

0.33 0.32 0.37 0.38 0.35

0.29 0.38 0.38 0.35 0.35 n.s. 0.05

0.34 0.35 0.39 0.35 0.36


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and inorganic fertilizer was supplied except P2O5. Third, the soil had an initially high P content. Previously, it has been shown that high P content in the soil strongly reduce the mycorrhizal infection.[15] The area in which the experiment was carried out had a level of P much more above the critical level. As known, in open field condition, 100 kg P2O5 ha21 is a sufficient amount of P2O5 for most of the plant. Since useful P2O5 level is high, responses of plants to mycorrhizae and P2O5 could not be determined. Native mycorrhizae or pure mycorrhizae adding may not contribute to growth or sometimes show negative effect under excessive P2O5 condition.[16] Plants growing on fertile soil often have mycorrhizae that less developed than those plants growing on non-fertile soils.[17] In both years, plant P content was determined above 0.20%[18] which is over critical value for garlic. In low level (0 kg ha21) P2O5 application, AM adding increased % P content of plant in both years, but it was not statistically important. Applied P2O5 doses were likely ineffective on plant growth and quality because there was a sufficient amount of P2O5 for garlic in soil and after certain threshold, addition of P2O5 does not affect plant growth and other parameters. Hence, any important difference was not found between control block and applications. In the same soil series in low P content area, garlic plant was grown in fumigated and AM added soil with control of soil fumigated without AM in the Department of Soil Science Research Area and garlic was found as extremely responsive to AM.[19] Garlic showed an excellent response to AM inoculation in either fumigated or non-fumigated conditions. From these results in the same soil series with the different P content, plant response to mycorrhizal formation varied. Under high P application and P accumulation, although mycorrhizal inoculation still is functioning but there is no more contribution to the plant development. It may be better to isolate the indigenous mycorrhizae at the same soil and re-inoculate for better P utilization. As a result, determination of AM species and concentration for plant are very important subject. In addition to this, determination of P2O5 content of research area and according to result of analysis, to determine a fertilizing program without damaging plant and environmental health is another important matter. Therefore, experiment should be repeated in low level of P2O5 application with several mycorrhizal species.

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