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Effects of akronin on growth of Rhizobium and yield of mung bean

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  • Save International Journal of Plant Research 2012, 2(6): 195-198 DOI: 10.5923/j.plant.20120206.04 Influence of Aclonifen on the Growth of Rhizobium Phaseolii and the Yield of Green Beans (Phaseolus Vulgaris L.) Orhan Vedat Gürsoy1, Hüseyin Padem2,* 1M ugla University, M ilas Vocational College, Department of Technical Programing, 48200, M ugla, Turkey 2International Burch University, Faculty of Engineering, Department of Genetics and Bioengineering, Francuske revoluciye, 71000, Sarajevo, Bosnia and Herzegovina Abstract In this study, the degree of the negative effect of Aclonifen containing herbicide on Rhizobium phaseolii, total mesophilic bacteria (TAM B), yeast and molds (YM ), the yield and correlation among parameters of bean under natural field conditions were investigated. Rhizobium phaseolii stock culture (8.71 log cfu/g), was mixed with the media in pots homogenously at a dose of 0, 1 and 2 g. When the young bean plants reached the 5-6 true leaf stage 600 g/l Aclonifen containing Challenge 600, was applied as a herbicide at the dose of 0, 625, 1250, 1875 and 2500 ml/ha, respectively. The effect of the Rhizobium and herbicide t reatments on Rhizobium phaseolii, TAMB, YM and the bean yield were tested. The results obtained fro m the trial revealed that the number of Rhizobium bacteria, TAM B, YM and also the yield were reduced by the increased herbicide dose. The number of TAMB and YM were not affected by Rhizobium treatments but yield was. Keywords Aclonifen, Mesophilic Bacteria, Mo lds, Rhizobium phaseolii, Yeast 1. Introduction Beans (Phaseolus vulgaris L.), with their h igh percentage in the share of vegetable production in Horticultura l act ivity, play a great role in many countries. Even despite great variations in the climate conditions, they can be grown all over, especially in semiarid countries[19]. In Turkey, they occupy an area of 98.2 hectares and 154.000 tons of dried beans and 563.000 ton fresh beans are produced per year[3]. The crop can be consumed either as a fresh vegetable when the pods are green or when the seeds are comp letely matured. Dried beans are a rich source of protein and agronomists desire to maximize their production. Since snap beans are produced mainly under irrigated conditions together with manure applicat ion to the fields, this may cause intensive weed emergence and growth. Then hand hoeing and the other means of mechanical controls are practiced. During the growth of the plants, the amount and activity of nitrogen fixing bacteria is very important and, in addition to weeds, are one of the most important pests which must be controlled. In dry bean production, Imazamo x shows potential as a * Corresponding author: (Hüseyin Padem) Published online at Copyright © 2012 Scientific & Academic Publishing. All Rights Reserved post-emergence weed control[6]. At low densities, either mechanical tillage or herbicides alone were effect ive but at higher densities herbicides combined with mechanical tillage were required for effective control[2]. Pre-plant incorporated and pre-emergence application of Imazethapyr alone or in a tank mixtu re with S-metolachlo r at low and high rates did not have any significant effect on plant height, dry weight, seed moisture content or yield but caused crop injury. The higher rates caused higher crop injury both alone or in tank mix application. PPI ap lication caused less crop in jury than pre-emergence applicat ion[16]. Imazamo x +fo mesafen mixture caused significant visual inju ry and tended to decrease lima bean height and yield. Despite some initial in jury observed in the metolachlor, Imazethapyr applied pre at 75 g/ha and quizalofop-p applied post at 72 g/ha have excellant potential as weed manage ment tools[13]. Herbicides registered for Lima Beans (Phaseolus lunatus L.) do not constantly control many troublesome weeds. Some herbicides registered for soybeans (Glycine max) will control these weeds but their tolerance in Lima beans is not known. Cloransulam, flu metsulan, metolach lor, sulfentrazo ne, lactofen, imazergapyr applied alone or in a mixtu re, low crop injury observed with cloransulam and Imazethapyr plus metolachlor. All others caused some injury but lactofen was the highest[5]. Many researchers did not pay too much attention to nitrogen fixing bacteria. During the selectivity tests of new compounds, many points have been taken into consideration. However, at the 196 Orhan Vedat Gürsoy et al.: Influence of Aclonifen on the Growth of Rhizobium Phaseolii and the Yield of Green Beans (Phaseolus Vulgaris L.) beginning of the field application herbicides will be absorbed by weed roots as well as crop roots. Though some slight stress may be observed in the crops even if the herbicide is accepted as selective, this situation could be neglected since the crops may recover quickly. Either light or mediu m damage may cause some delay in the growth of the crop, too. Earlier studies have demonstrated the adverse effects of some kinds of herbicides on Rhizobium growth and its symbiosis. Insufficient information is availab le on the effect of herbicides on Rhizobium, total mesophilic bacteria, yeast and moulds on some legumes. enumerated on Plate Count agar (Merck, Darmstadt, Germany). The number of yeasts and molds (YM ) was determined in Potato Dext rose agar (Merck, Darmstadt, Germany) reduced to pH 3.5 with tartaric acid. Rhizobium phaseolii counts of the soil samples were enu merated on Mannitol Yeast Ext ract agar and incubated at 25℃ for 3-4 d ays [17 ]. The microbia l counts were determined as colony forming units (cfu) in gram of the samp les. The results of microbio logical analyses shown as log cfu/g. All analyses were formed in duplicate. 2. Main Body 2.1. Materials and Methods The study was carried out at Suleyman Demirel University Agricultural Research and Experimental Station. The 18 liter capacity plastic bags were filled with the med ia prepared as a mixture of manure, loamy soil and sand at a ratio of 1:1:1[15]. at a total of 15 liters of each. In order to provide the natural environ ment, the filled bags were buried in the field to keep both the soil and plastic bags surface at the same level. Before the applicat ion of Rhizobium phaseolii, the microbio logical p roperties of the media were determined as total mesophilic bacteria, (TAMB), 6.63 log cfu/g, yeast and molds (YM): 4.30 log cfu/g and Rhizobium phaseolii 1.40 log cfu/g. Rhizobium phaseolii stock culture (8.71 log cfu/g), produced by Soil and Fertilizer Research Institute in Ankara-Turkey, was mixed with the med ia in pots homogenously at a dose of 0, 1 and 2 g. Two seeds of the Ro mano bean variety were sown in each pot on June 5th. This variety of beans is widely produced in Turkey for fresh bean consumption. The planting distance was adjusted to 50 cm between rows and 25 cm between plants. When the young bean plants reached the 5-6 true leaf stage[4]. 600 g/l Aclonifen containing Challenge 600 were applied as a herbicide at the dose of 0, 625, 1250, 1875 and 2500 ml/ha, for the first time. The surface area of the pots was 962 cm2 The generally reco mmended dose should have been 1250 ml/ha. The field trial[8] was designed and carried out according to split plot design with four rep licat ions. Each rep licat ion consisted of four pots and 8 plants. During the growing period with 30 days interval three times 20 g soil samp les were taken fro m each pot starting fro m 17th of July in o rder to determine the quantity of Rhizobium, TAMB and YM. Each t ime the samples were taken in the morning 24 hours after each irrigation. 2.2. Microbi ological Analyses The sterilized jars were used for the soil samples. The 10 g of soil samples mixed with various concentrations of Rhizobium phaseolii and herbicide were added in 90 ml NaCl solution (0.85 %) and diluted up to 10-5. The total aerobic mesophilic bacteria (TAMB) were 2.3. Fi ndings 2.3.1. The Effects of Aclonifen Treatments on R. Phaseolii Bean plants grown with different doses of herbicide and R. phaseolii stock culture based on the results shown on the Table 1. Rates of Aclonifen adversely affected the R. phaselii counts. It was determined that the number of Rhizobium bacteria in the med ia were reduced with the increased doses of herbicides. The interactions of HerbicidexRhizobium were significant at the 0.01 level. At the same time the t wo doses of Rhizobium were not significant between themselves but the control was significantly different. Table 1. Rhizobium phaseolii counts (log cfu/g) of the soil samples Rhizobium sp Herbicide rat es 0 g/pot Rhizobium 1 g/pot Rhizobium 2 g/pot Rhizobium means 0 ml/ha 625 ml/ha 1250 ml/ha 1875 ml/ha 2500 ml/ha Average 246.3 99.6 96.3 67.3 67.6 1.15 b 702 448.3 349.3 183 92.6 3.55 a 711 436 352 195.6 93 3.57 a 5.53 a * 3.27 b 2.65 c 1.48 d 0.84 e * Means with different letters within columns are significantly different (p<0.05) 2.3.2. The Effects of Treatments on TAMB Table 2. T AMB counts (log cfu/g) of the soil samples Herbicide rat es 0 g/pot Rhizobium TAMB 1 g/pot Rhizobium 2 g/pot Rhizobium means 0 ml/ha 625 ml/ha 1250 ml/ha 1875ml/ha 2500 ml/ha Average 690 409.6 363.3 295.3 220.3 3.95 a 666 404 351.6 273 207.6 3.80 a 629 363.6 345 252 195.3 3.56 a 6.61 a * 3.92 b 3.53 c 2.73 d 2.07 e * Means with different letters within columns are significantly different (p<0.05) As can be seen from Table 2, the number o f TAMB was not affected by the Rhizobium treatments. On the other hand, the herbicide treat ments were statistically effective on the number o f TAM B. Increased herbicide doses reduced the number o f the TAMB. HerbicidexRhizobium interreaction International Journal of Plant Research 2012, 2(6): 195-198 197 was not significantly important at the level of 0.05. 2.3.3. The Effect of Treat ments on YM It is statistically determined that the number of YM was not affected by the Rhizobium treatments. However the effect of the interreaction of the HerbicidexRhizobium on YM nu mbers in med ia was statistically significant at the 0.01 level. The nu mber of YM was high on non-treated check pots but it was the lowest on the fourth dose of the herbicide application. It is clear that increased herbicide dose reduced the number of YM in the media. Table 3. YM counts (log cfu/g) of the soil samples Herbicide rat es 0 g/pot Rhizobium YM 1 g/pot Rhizobium 2 g/pot Means Rhizobium 0 ml/ha 625 ml/ha 1250 ml/ha 1875 ml/ha 2500 ml/ha Average 488.6 309.6 275.6 216 163 2.90 a 511.3 302.3 262 194.3 132.3 2.80 a 497 286 245.6 191 105.3 2.64 a 4.98a* 2.99 b 2.61 c 2.00 d 1.33 e * Means with different letters within columns are significantly di fferent (p<0.05) Table 4. The effect of herbicide and Rhizobium treatments on yield (g/p lant ) Herbicide rat es 0 g/pot Rhizobium Yield 1 g/pot Rhizobium 2 g/pot Rhizobium Means 0 ml/ha 625 ml/ha 1250 ml/ha 1875 ml/ha 2500 ml/ha Average 681 574 493 462 416 525 c 827 777 709 643 562 703 a 814 783 653 572 559 676 b 774 a * 711 b 618 c 559 d 512 e * Means with different letters within columns are significantly di fferent (p<0.05) Table 5. Correlation among parameters Yield TAMB Rhizobium YM Yield 1.00 TAMB 0.65+-0.12 *** 1.00 Rhizobium 0.91+-0.06 *** 0.71+-0.11 *** 1.00 YM 0.66+-0.11 0.98+-0.03 0.73+-0.10 *** *** *** 1.00 ***All of the correl ations among the parameters taken into account in the trial were signi ficant at 0.01 level 2.3.4. The Effects of Treatments on Yield and Correlat ion among Parameters The highest yield was obtained from the 1 g/pot Rhizobium treatment. This was followed by the 2 g/pot treatment and the lowest yield was obtained from the check pots. As can be clearly seen fro m the table that the herbicide application reduced the yield. The h ighest yield was obtained fro m the check pots. The differences of either herbicides or Rhizobium application were significant at 0.05 level. A lso the effect of interreaction between Herbicidex Rhizobium was significant on the yield. 2.4. Results and Discussion Since herbicides are necessary to achieve maximu m y ield, their in fluence on nodulation may conflict with the crop managements[10]. Herbicide applicat ion is very co mmon in bean production. It is well known that the bean plants can get most of their nitrogen needs main ly by the nodules formed on their roots fro m the n itrogen in the at mosphere. Many of the chemicals, e.g. pesticides, may be potentially hazardous and associated with sy mbiotic nit rogen-fixing microorganis ms[12]. The inoculated seed has to provide the incoming rhizosphere with enough microorganisms to nodulate and fix nitrogen[14]. Based on the results obtained from this trial, it was found that the application of aclonifen had a reducing effect on Rhizobium bacteria together with TAMB and YM. There is a probability that Aclonifen may have a to xic effect on Rhizobium, TAMB and YM population existing in the bean producing field. To xicity of Aclonifen to Rhizobium, TAMB and YM increased progressively with increase in rates of herbicide. Similar results were obtained by Ahemad and Khan (2009) that quizalafop-p-ethyl and clodinafop proved to have a lethal effect on symbiot ic properties. Weeds cause an 8.7 % reduction in bean production in the USA[7]. The severe co mpetit ion for nutrients light and water between crops and weeds at early stage requires pre-emergence herbicide application. A mong the many herbicides used for weed control in green bean crops, Aclonifen has proved to be effective against a wide spectrum of broad-leaved weeds in legu mes[18]. Co mpared to check pots, the population of Rhizobium increased in the Rhizobium applied pots. However there were no differences in terms of Rhizobium population between 1 g/pot and 2 g/pot Rhizobium application. Also Rhizobium application had no effect neither on the TAMB nor YM populations. The infective phase of the symbiosis begins just before the contact occurs between bacteria and root hairs[9] and during this period, the association is highly sensitive to the soil environment[11]. 3. Conclusions Both the Rhizobium and herbicide application considerably affected the bean yield. The highest yield was obtained from 1 g/pot Rhizobium application. Increased herbicide application reduced the yield. Very important correlations were determined among all the parameters evaluated in this research. These findings are probably due to the quick inactivation of Aclonifen in growing media. As a result of this research, it was found out that the application of aclonifen for controlling weeds in bean production had a negative effect on the soil microbiology. The application of Aclonifen may only be recommended as a last resort for weed control in bean production. The trial was 198 Orhan Vedat Gürsoy et al.: Influence of Aclonifen on the Growth of Rhizobium Phaseolii and the Yield of Green Beans (Phaseolus Vulgaris L.) carried out under natural field conditions. The same type of trial may also be carried out in a laboratory to find out the effect of Aclonifen on the soil microbial activ ity. [9] R. F. Fisher and S. R. Long (1992). Rhizobium-plant signal Exchange. Nature, 357:655-659. [10] N. Gonzalez, J. J. Eyherabide, M . I. Barcelonna, A. Gaspari and S. Sanmartino (1999). Effect of soil interactingherbicides on soybean nodulation in Balcarce, Argentina. Pesq. Agropect. Bras., Brasilia, 34(7):1167-1173. REFERENCES [1] M . Ahemad and M . S. Khan, (2009). Toxicity Assessment of Herbicides Quizalafop-p-Ethyl and Clodinafop towards Rhizobium pea symbiosis. Bull. Environ Contam. Toxicol. 82:761-766. [2] Amador-Ramirez, M . D., Wilson, R. G.& M artin, A. R., (2001) Weed control and dry bean (Phaseolus vulgaris) response to in-row cultivation, rotary hoeing, and herbicides. Weed Technology, 15(3):429-436. [3] Anonymous (2009). Devlet Istatistik Kurumu. The Summary of Agricultural Statistics. ( o?tb_id=45&ust_id=13).15.12.2009. [4] Anonymous (2010). Yabancı Ot Standart Deneme M etotları/2009. art_ilac/yosidm.pdf. 18.01.2010. [5] W. A. Bailey, H. Wilson and T. Hines (2003). Weed control and snap bean (Phaseolus vulgaris) responce to reduced rate of fomesafen. Weed Technology, 17.(2):269-275. [6] R. E. Blackshaw, L. J. M olnar, H. M uendel, G. Saindon and X. Li (2000). Integration of cropping practices and herbicides improves weed manegement in dry bean (Phaseolus vulgaris). Weed Technology. 14(2):327- 336. [7] H. H. Cramer (1967). Plant Protection and World Crop Production. Bayer, Planzeschutz, Leverkusen, 3-524. [8] O. Düzgünes, T. Kesici, O. Kavuncu and F. Gürbüz, (1987). Arastırma Denmeme M etotları. Ankara University Publishing No: 1021. Ankara. [11] H. H. Keyser, P. Somasegaran and B. B. Boholool (1993). Rhizobial ecology and technology. In M etting Junior, B. (ed). Soil microbial ecology. New York: M arcel Dekker, 205-226. [12] S. Khan, A. Zaidi and M . Aamil (2004). Influence of herbicides on Chickpea-M esorhizobius symbiosis. Agronomie. 24:123-127. [13] K.. M cNaughton, P. H. Sikkema and D. E. Robinson (2004). Herbicide tolerance of Lima Bean (Phaseolus lunatus) in Ontario. Weed Technology. 18(1):106-110. [14] R. J. Rennie and S. Dubetz (1984). Effect of fungicides and herbicides on nodulation and N2 fixation in soybean fields lacking indigenious Rhizobium japonicum. Agronomy Journal, 76: 451-454. [15] A. Sevgican (1999). Topraksız tarım, Örtü Altı Sebzeciliği, Cilt 2. Ege University Publishing, No: 626. Bornova İzmir. [16] N. Soltani, C. Shropshire, T. Cowan and P Sikkema (2004). Tolerance of black beans (Phaseolus vulgaris) to soil application of S-metolachlor and Imazethapyr. Weed Technology. 18(1):111-118. [17] D. L. Sparks and J. M . Bartel (1996). (Ed), M ethods of Soil Analysis: Chemical M ethods Part 3’’ SSSA. [18] M . J. Vangessel, D. V. M onks and Q. R. Johnson (2000). Herbicides for potential use in Lima Bean (Phaseolus lunatus) production. Weed Technology. 14(2):279-286. [19] H. Vural, D. Esiyok and I. Duman (2000). Kültür Sebzeleri (Sebze Yetistirme). Ege Üniversitesi, Ziraat Fakültesi, Bahçe Bitkileri Bölümü, Ege Üniversitesi Basım Evi, İzmir.

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