WTfi ‘- A COMPARISON OF THE EFFECTS 0? TWO COPPER CARRIERS ON THE YEELD AND COPPER CONTENT OF SEVERAL CROPS AND THE EFFECTS OF CONTACT APPLXCATION OF SEVERAL MINOR ELEMENT CARRIERS ON THE STAI‘JD AND YIELD 3F ONEONB THESiS FOR ‘7'?le DEGREE O? 51.5,. MICHIGAN STATE. COLLEGE LAWRENCE N. SHEPHERD $95.5 This is to certify that the thesis entitled A Comparison of Two Copper Carriers on the Yield and Capper Content of Grape and the Effect of Several Minor Element Carriers on the Yield and Stand of Onions presented by Lawrence Shepherd has been accepted towards fulfillment of the requirements for “Ct 31" 8 degree in 861 m C. Date Ma 11 A COMPARISON OF THE EFFECTS OF TWO COPPER CARRIERS ON THE YIELD AND COPPER CONTENT OF SEVERAL CROPS AND THE EFFECTS OF CONTACT APPLICATION OF SEVERAL MINOR ELEMENT CARRIERS ON THE STAND AND YIELD OF ONIONS BY LAWR ENCE N. SHEPHERD ad. A. THESIS Submitted to the School of Graduate Studies of Michigan State College of Agriculture and Applied Science in partial fulfillment of the requirements for the degree of MASTER OF SCIENCE Department of Soil Science 1955 THESIS \. ABSTRACT Field studies were conducted at the Michigan Muck Experi- mental Farm to investigate the comparative effects of Calumet Brown Copper Oxide and c0pper sulfate as measured by the yield reSponse and copper content of sudan grass, potatoes, carrots, and onions. Two rates of molybdenum were applied to measure yield reSponse of Sudan grass to this element. A. further study was conducted with six minor element carriers placed in contact with the seed to meas-‘ ure their effects on the stand and yield of onions. The six carriers used were: (1) copper sulfate (25 percent copper); (Z) Calumet Brown Copper Oxide (50 percent copper); (3) manganese sulfate (24 percent manganese); (4) NU M (a neutral compound containing 41 percent manganese); (5) zinc sulfate (36 percent zinc); and (6) NU Z (a neu- tral compound containing 55 percent zinc). The findings of the studies were as follows: 1. Sudan grass did not respond significantly in yield to the application of ammonium molybdate. 2. In the first year of copper application to a virgin, copper- deficient muck, the response of sudan grass and carrots to 25 pounds ii 354792 of copper oxide per acre was significantly greater than when copper sulfate was applied at the same rate of total copper. 3. Very little difference in yield occurred between the two rates of copper applied or the two copper carriers in the second year; although in general the crops responded significantly to a cop- per application. 4. More copper per acre was removed in the cr0ps from the copper-oxide treated plots than from plots receiving the same amount of copper in the form of copper sulfate. 5. The 3-1/8 pounds per acre rate of copper applied for two successive years met the needs of the crops since no response in yield was obtained from higher applications. However, the copper content of plant tissue was slightly higher at the higher rate of copper application. 6. Copper sulfate at' 25 or 50 pounds per acre placed in contact with the seed significantly decreased the stand and yield of onions, whereas copper oxide applied at equivalent rates of copper in contact with the seed of onions was not detrimental. 7. Manganese sulfate when applied at 200 pounds per acre in contact ‘with the seed of onions significantly decreased the stand and yield; however, the 100 pounds per acre rate was not harmful. iii 8. Other minor element carriers applied either in combina— tion or alone and which had no harmful effects when placed in con- tact with the seed of onions were: (1) NU M at 60 and 120 pounds per acre; (2) NU Z at 15 pounds per acre; and (3) zinc sulfate at 15 pounds per acre. iv ACKNOWLEDGMENTS The author expresses his sincere gratitude to the members of the Soil Science Department staff and his fellow graduate students for their fine spirit of co-operation and helpful suggestions in the inter— est of this investigation. Special appreciation is directed to Dr. J. F. Davis for his initiation of the experiments, his guidance, timely assistance, and advice throughout this study. He is also indebted to Dr. R. L. Cook, Dr. L. M. Turk, Dr. K. Lawton, and others for helpful suggestions, encouragement, and assistance in completing this work. The writer is grateful for the untiring help given and for the use of equipment furnished by Dr. E. J. Benne and associates of the Agricultural Chemistry Department. The financial support given this project by Calumet and Hecla, Incorporated, Calumet, Michigan, was very deeply appreciated by the autho r . Experiment Experiment Experiment TABLE OF CONT ENT S I... ............................ II . ............................. III .............................. Preparation of Samples ....... . ................. Analytical Procedure ......................... R ESUL TS AND DISCUSSION ....................... Experiment I ............................... Sudan grass .............................. Experiment 11 .............................. Sudan grass .............................. Onions Carrots Potatoes vi 10 12 13 l4 16 18 18 22 22 26 26 28 Deficiency symptoms ....................... Copper removal ........................... Experiment III .............................. Onions ................................. vii LIST OF TABLES TABLE III. IV. VI. VII. The Effects of Two Rates of Ammonium Molybdate and Two Copper Carriers on the Yield and Copper Content of Sudan Grass ..... The Effect of Two Rates of Application of Copper Oxide and Copper Sulfate on the Yield and Copper Content of Sudan Grass and Onions .......................... The Effect of Two Rates of Application of Copper Oxide and Copper Sulfate on the Yield and Copper Content of Carrots ........ The Effect of Two Rates of Application of Copper Oxide and Copper Sulfate on the Yield and Copper Content of Potatoes ........ Copper Removed per Acre by Sudan Grass, Potatoes, Onions, and Carrots ............. The Effect of Rates of Application of Several Minor Element Carriers Placed in Contact with the Seed on the Stand and Yield of Onions (1953) ............... The Effect of Rates of Application of Several Minor Element Carriers Placed in Contact with the Seed on the Stand and Yield of Onions (1954) ............... viii Page 21 25 27 29 32 37 39 LIST OF FIGURES FIGURE Page 1. Photograph Showing General View of the Plots in Experiment I ...................... 20 2. An Over-A11 View of the Plots in Experi- ment 11 Showing the Onions, Carrots, and Potatoes During the 1954 Season .............. 24 3. Color Difference Caused by Copper ............ 31 4. Fertilizer Placement Drill .................. 36 ix INTRODUCTION Most crops may respond to copper added as a fertilizer con- stituent when grown on certain organic soils in Michigan. Not only does the same crop differ in degree of response when grown on dif- ferent soils, but crops and even varieties of the same crop often differ in their response to this element. In most of the former Michigan studies involving the c0pper nutrition of plants, the copper was supplied as copper sulfate. Sev- eral other materials containing copper (4) have been shown to supply plant needs. However, they are not recommended in Michigan at the present time. With the introduction of a fertilizer grade of copper oxide, it seemed advisable to make comparative studies to determine the value of this material as a source of copper for plants. The- initial work at Michigan State College was completed by Johnson (14) and van Eck (23) under greenhouse conditions. The purpose of this study was to investigate under field con— ditions the comparative effectiveness of copper sulfate and Calumet Brown Copper Oxide as measured by yield response and copper con- tent of several crops. A further field study was undertaken to determine the effect of several minor element carriers placed in contact with the seed on the stand and yield of onions. The experiments were located at the Michigan Muck ExPeri- mental Farm near East Lansing. REVIEW OF LITERATURE Recent reviews of the literature on copper have been com- pleted by several Michigan investigators (4, l4, l7, I9, 23). Such topics as the essential nature of copper in plant and animal metabo- lism, the physiological role of copper in plants, copper in the soil, and symptoms of copper deficiency in plants were extensively cov- ered. For other detailed reviews on copper, the publications of Sommer (21) and Elvehjem (10) were fairly complete. The interest of this review was primarily that of factors affecting the response of plants to copper and copper materials used as fertilizer. Copper was first shown to be essential for plants by Som- mer in 1931 (20); also later in the same year, Lipman and McKinney (16) gave further proof of its essentiality to plants. These authors used purified reagents and water redistilled from pyrex in nutrient cultures to demonstrate the essential nature of copper for the growth of plants. Hill and Bryan (13), working with sandy soils collected from areas of virgin Florida soils known to be inherently poor and on which "salt sickness" of cattle was known to occur, found that c0pper had a nutritive value in the growth of mustard over a wide range of soil types. Their results also indicated that copper may be deficient on certain soil types in a manner similar to that of potash and phosphate. Rehling and Truog (18), in experiments with corn, tomatoes, and sunflower plants grown in nutrient solutions with a carbonated water extract of Milorganite as a source of minor elements, showed that the Milorganite extract significantly increased the dry weight of plants and their content of copper and other minor elements. Field studies conducted by Harmer (11), on organic soils using copper sulfate as a copper carrier, showed that the amount of re5ponse to copper nnay depend on the crop species, on the re- action of the plowed layer and of the drained underlying layers, on the extent of soil drainage, and on the seasonal climate. Harmer concluded in general that most crops were benefited by copper if the natural pH of the soil was 6.0 or less; the more reSponsive crops showed a need for c0pper if the soil pH was as high as 6.5; and the most responsive cr0ps appeared to respond in yield under alkaline soil conditions. He found that the amount of re5ponse shown by a responsive crop was greater in a hot, dry summer than in a cool, wet one and that copper improved the color and flavor of some crops. Armiger 9121. (I) compared copper sulfate and a spent cop- per pyrophosphate catalyst as a source of copper for shallu and winter oats grown in the greenhouse on copper-deficient soil. They found that the copper content and the growth response of the crops to spent copper pyrophOSphate catalyst was as high as that found with copper sulfate. Harris (12) found oats to be very sensitive to a lack of c0p- per, and the deficiency was corrected by applying ten pounds of copper chloride per acre. Very small amounts of c0pper applied to the soil had considerable residual effect, and appeared to have an effect on the resistance of oats to cold. Berger and Truog (3) increased the yield of sweet corn by the application to a Miami silt loam of five to ten pounds of copper sulfate per acre. The increases ranged from 5 to 40 percent over the check. However, yield increases were not obtained from copper applied to Carrington silt loam, a Prairie soil high in organic matter. Brown and Harmer (5) reported that several copper carriers (copper sulfate, tetra copper calcium oxychloride, copper hydroxide, c0pper pyrophosphate catalysts, and C. P. electrolytic c0pper) when applied to organic soil markedly increased the yield of wheat, pre- vented or corrected copper-deficiency symptoms, and reduced concentrations of various plant-food elements in the tissue. Slightly lower crop yields and a lesser reduction in the amount of the ele- ments in the tissue were obtained from soil receiving metallic cop- per than from soil treated with the other carriers, suggesting that a determination of such concentrations might be a measure of the efficiency of the copper carriers. The absorption of applied copper by the crops investigated affected the assimilation of nitrogen, phos- phorus, potassium, calcium, magnesium, silica, and iron. Steenbjerg and Boken (22) found that certain factors influence the degree of c0pper deficiency. The factors were: pH value, soil type (humus content), the nature of the previous crop, and the addi- tion of clay and sand, et cetera. The results of pot and field studies with different copper minerals (copper pyrites, bornite, copper glance, cuprite, and malachite) and copper chemicals (copper sulfate, copper oxide, and metallic copper) showed that several of the slightly solu- ble copper compounds were suitable as copper fertilizers if ground to a suitable fineness. In addition, their residual effect, like that of copper sulfate, was considerable. Johnson (14), in making a comparative study between copper oxide and copper sulfate under greenhouse conditions, found there was little or no difference between the effectiveness of either c0pper oxide or copper sulfate in increasing the yield and copper content of spinach. or Sudan grass when both carriers were used at the same rate of total copper per acre. Kretschmer and Forsee (15) showed copper oxide to be as effective as copper sulfate in increasing crop yields. van Eck (23), working in the greenhouse with Sudan grass, spinach, and head lettuce, found copper oxide as effective as copper sulfate in correcting copper deficiency and increasing the yield and copper content of the three crops. EXPERIMENTAL PROCEDURE Introduction The three experiments reported herein are field studies ini- tiated by Dr. J. F. Davis. The plots were located at the Michigan Muck Experimental Farm near East Lansing. The soil, a copper-deficient Houghton muck, was analyzed by Brown (4). It ranged in pH from 6.0 to 6.3, and contained 86 per- cent organic matter, 3.3 percent total nitrogen, 0.21 percent potas- sium, 0.12 percent phosphorus, 2.5 percent calcium, 0.27 percent magnesium, 1.3 percent iron, and 0.0011 percent c0pper. Experiment I was initiated to measure the effect of two rates of ammonium molybdate (3 and 6 pounds per acre) on crop yields and to compare the effects of copper sulfate and copper oxide on the yield and copper content of the crops grown. Experiment 11 was arranged to study the comparative effects of rates of application of copper sulfate and c0pper oxide on the yield and cepper content of several crops. The object of Experiment III was to determine the effects of rates of application of several minor element carriers placed in con— tact with the seed on the stand and yield of onions. 8 Experiment I This experiment was initiated in 1950 to measure the effect of two rates of ammonium molybdate on the yield of Sudan grass and to compare the effects of copper sulfate and cOpper oxide on the yield and copper content of the crop. The plots were arranged as a randomized block design with three replications and five treat- ments. Three years' yield data were obtained for Sudan grass be- fore the experiment was discontinued in 1954. The 1953 crop was analyzed for c0pper. Treatments were as follows: (1) no copper; (2) 2.5 pounds of copper per acre applied in the sulfate form; (3) 2.5 pounds applied in the oxide form; (4) 2.5 pounds applied in the sulfate form with 6 pounds of ammonium molybdate; (5) 2.5 pounds applied in the sulfate form with 3 pounds of ammonium molybdate. The copper treatments were made in 1950, 1951, and 1952, and the ammonium molybdate treatments in 1950, 1951, 1952, and 1953. A basic application of 1,000 pounds of 0-10-30 per acre was made annually on all plots. The minor element carriers were mixed with a convenient volume of soil and broadcast by hand. The Sudan grass was planted the last week in May and har- vested the last part of September each year. Samples of Sudan grass 10 were taken at the time of harvest in 1953 for analysis of the copper content. Expe rime nt 11 In 1952 an area of virgin organic soil was cleared and tile drain installed to prepare an area for further field studies of the comparative effects of rates of application of copper oxide and cop- per sulfate on several crops. The plot design was a randomized block consisting of five treatments replicated four times. Three crops were grown each year. The five treatments were as follows: (1) check (no copper); (2) 3.125 pounds of c0pper per acre applied as copper sulfate; (3) 3.125 pounds applied as Calumet Brown Copper Oxide; (4) 12.5 pounds applied as copper sulfate; (5) 12.5 pounds applied as Calumet Brown Copper Oxide. The indicator crops used in 1953 were Sudan grass, Sebago potatoes, and Chantenay carrots. In 1954 Sudan grass was replaced by Downing's Yellow Globe onions, and the potatoes and carrots were continued for the second year. The data of two years' yield were obtained for Sudan grass and onions. Harvesttime samples were tested for content of c0pper. 11 An application of 1,000 pounds per acre annually of a basic fertilizer consisting of ammonium nitrate, treble superphosphate and potassium chloride was applied to each cr0p. Sudan grass and pota- toes received an 0-10-30, and carrots and onions received a 5—10-20. The basic fertilizer for the potato crop was applied with the grain drill before planting. The copper was weighed on a torsion balance for each plot, mixed thoroughly with a volume of screened muck, and broadcast by hand. For the other crops, the fertilizers were banded two inches below the seed at the time of planting. The copper carriers were mixed in the basic fertilizer according to plot requirement. The 1953 planting date for all three crops was May 29. Har- vest dates in 1953 were: potatoes, October 29; carrots, October 15; and Sudan grass, September 22. In 1954 planting dates were April 26 for onions and carrots, and May 20 for potatoes. Harvest dates in 1954 were: potatoes, October 22; onions, September 17; and car- rots, October 18. All plots received several Spray applications of manganese throughout the growing season. Also, a normal spraying program was carried out for control of diseases and insects. 12 Expe riment III In this experiment, six minor element carriers were applied separately and in combination at several rates. In all cases the minor element carriers were placed in contact with the seed to measure their effects on the stand and yield of onions. The carriers used were: (1) copper sulfate (25 percent cop- per); (2) Calumet Brown Copper Oxide (50 percent copper); (3) man- ganese sulfate (23.65 percent manganese); (4) NU M (a neutral com- pound containing 41 percent manganese); (5) zinc sulfate (36 percent zinc); and (6) NU Z (a neutral compound containing 55 percent zinc). In 1953, the experiment contained nineteen treatments with two replications. In 1954, the experiment was changed to include three replications and twenty-four treatments. The treatments ap— pear in Tables VI and VII of the experimental results. The varieties of onions were Baldwin's Early Yellow Globe in 1953 and Downing's Yellow Globe in 1954. A basic application of 5-10-20 at 1,000 pounds an acre was made on all plots. The fertilizer was placed 2 inches below the seed at planting time. Planting dates were June 2 in 1953 and May 4 in 1954. Harvest dates were September 30 in 1953 and September 17 in 1954. 13 Minor elements had been applied in previous years to the area used for this experiment. Therefore, very little response in yield was expected from minor elements applied where a residual effect of the element carried over from previous treatments. How- ever, it was assumed that if a particular carrier placed in contact with the seed was toxic, a depression in yield or stand should fol- low. P reparation of Sample 5 All plant samples were obtained from mature crops at har- vest time. Plant parts analyzed for copper content were the potato tuber, the carrot tap root, the onion bulb, and the above-ground portion of Sudan grass including any seeds that may have been pres— ent on the plants. The potatoes, carrots, and onions were sliced thin with a vegetable slicer, placed in paper bags, and dried in a forced-draft electrical oven at 174 degrees Fahrenheit. Plant material was ground in a Wiley Mill through a 20 mesh screen and stored in 8-ounce wide-mouth sample jars. Separate samples for each crop and each plot were taken at harvest time and prepared as stated above. A further sample was prepared by 14 bulking the replicates of each treatment for a crop. Before analysis, the samples were oven-dried at 180 degrees Fahrenheit to remove moisture adsorbed during grinding. Analytical Procedure (2, 6, 7, 8.) Known weights of plant tissue (3 to 5 grams) were placed in 250-mi11i1iter pyrex beakers and digested by using nitric acid and a 2:1 mixture of 70 percent perchloric acid and concentrated sulphuric acid. To the dry or nearly dry residue were added 3 milliliters of concentrated hydrochloric acid and 50 milli- liters of distilled water. The material was then heated just under the boiling temperature for about 15 minutes. The solution was filtered through No. 42 Whatman filter paper into 250-milli1iter volumetric flasks. The residue was washed with boiling distilled water until the filtrate was free of chlorides. The volumetric flasks were then cooled and the contents brought to volume with distilled water and mixed. Aliquots of 50 milliliters were placed in 125 cubic centimeter separatory funnels and shaken with 5 milliliters of 15 percent citric acid solution. The solution was made alkaline by adding 1:1 ammonium hydroxide, using neutral litmus paper as an indicator. Then, 10 15 milliliters of sodium-diethy1-dithiocarbamate reagent (1 gram per liter) were added and shaken. The copper carbamate complex was then extracted by shaking with 4-milliliter portions of carbon tetra— chloride for three minutes. The extraction was repeated three times. Each time the carbon tetrachloride layer was drawn off into 25- milliliter Erlenmeyer flasks. The extracted material was filtered through a small quantity of anhydrous sodium sulfate using No. 12 Whatman filter paper into 25-milli1iter volumetric flasks. The con- tents of the flasks were then brought up to volume with carbon tetrachloride. Light transmission was determined by a Cenco-Sheard-Sanford photelometer using a Corning glass light filter (number 554, lantern blue with maximum transmittance at 4500 A°) and l-centimeter ab- sorption cells. Analytical reagent carbon tetrachloride was used as the reference standard. Blank determinations were carried through the procedure in the same manner as described above to correct readings of the unknown samples. Copper content was evaluated from a curve relating transmittance and concentration. Quantitative measurements of copper content were checked by using reference samples furnished by the Agricultural Chemistry Department. R ESULTS AND DISCUSSION In the initial analysis of plant tissue for copper content, dry ashing in an electric muffle furnace was used. At first, duplicate analyses were made for each crop and each replication; however, such erratic results were obtained that it was necessary to investi- gate possible reasons for the variation. It was possible to repro- duce readings by rerunning a second aliquot from the same ash ex- tract; thus, it was assumed that the variation occurred in the ashing procedure. It was found that monel metal trays (monel metal contains about 28 percent copper) had been used in the muffle furnace of the Soils Department graduate research laboratory. When ashing in this muffle, the blanks often contained as high as six parts per mil- lion of copper. Ashing was then switched to muffles of the Agricultural Chem— istry Department. When using the muffle furnaces of the Agricul- tural Chemistry Department, the blank determinations were not af— fected; but variation often occurred between duplicates of the same sample. Variations of four parts per million in copper content of duplicates from the same sample were common. 16 17 More than three hundred tests were run using several tech- nique changes in the muffle furnace ashing; i.e., such techniques as treating material previously ashed overnight at 500°C with nitric acid, evaporating the nitric acid, and then reashing. Also, pretreat- ment with sulphuric acid before ashing was tried. Regardless of the techniques employed, variations often occurred between duplicate analyses of the same sample. Since Kretschmer and Forsee (15), using a wet digestion pro- cedure, reported copper content of plant tissue to tenths of a part per million and this investigator was unable to obtain agreement in the parts per million figure, it was decided to try a wet digestion procedure of ashing. A series of samples were run using nitric, and a 2:1 mixture of 70 percent perchloric and concentrated sulphuric acids for digestion of the plant tissue. In every case, no difference between the dupli- cates was observed. It was then decided to run all the samples of bulked replicates in duplicate using the wet digestion method of ash- ing. When duplicate test results varied one part per million, a re- run of the sample placed the copper content within two-tenths of a part per million of one of the original figures. After comparing the results obtained from the dry ashing in the electric muffle furnace with the wet digestion procedure, it was 18 decided that the results from the muffle furnace ashing were unsatis- factory. The reasons for the erratic and unsatisfactory results from the muffle furnace ashing are unknown. However, because of the close agreement obtained in results from replicated determinations by the wet digestion procedure and in view of the erratic results obtained from the muffle ashing, all of the results obtained from the dry ashing in the electric muffle furnace were discarded. The copper contents of plant tissues reported herein are from the results obtained from the samples of the bulked replicates run in duplicate and from the wet digestion method of ashing. Experiment 1 Sudan grass. Figure 1 shows a general view of the plots in Experiment I. The reduced growth of the Sudan grass on the no- copper plot in the foreground is especially apparent. The character- istic visual copper deficiency symptoms were present on the Sudan grass check plots. Table I shows that Sudan grass did not respond to the appli- cation of ammonium molybdate at either rate of application of molyb- denum. There was a trend toward depression of yield at the 6-pound rate of ammonium molybdate, but the difference was not significant. 19 FIGURE 1 PHOTOGRAPH SHOWING GENERAL VIEW OF THE PLOTS IN EXPERIMENT 1 (Notice in particular the reduced growth of the Sudan grass on the no-copper’ plot in the foreground.) 20 21 TABLE I THE EFFECTS OF TWO RATES OF AMMONIUM MOLYBDATE AND TWO COPPER CARRIERS ON THE YIELD AND COPPER CONTENT OF SUDAN GRASS Pounds Of Am- Percent Pounds momum Cop e p r f b- o MOIY Tons per Acre in the Coppe r C oppe r date Carrier Applied A lied Oven PP 1951 1952 1953 Avg. Dry per per , Tissue Acre Acre 1953 An- nually No copper . . . 0.0 0.0 8.2 2.9 1.7 4.2 0.00120 Copper sulfate . . . 2.5 0.0 10.2 11.4 6.1 9.2 0.00071 Copper oxide . . . . 2.5 0.0 8.6 10.6 6.5 8.6 0.00145 Copper sulfate . . . 2.5 6.0 8.0 9.5 5.6 7.7 0.00150 Copper sulfate . . . 2.5 3.0 8.8 11.7 7.8 9.1 0.00137 L.S.D. (5 percent level) . . . 1.5 2.7 2.5 22 Also, there was an indication of slight benefit from molybdenum at the low rate of application. The data show that the yield of Sudan grass was significantly increased by an application of copper, although there was no signifi- cant difference between yields of plots where the two carrierswere used. Although the copper content of the tissue differed with treat- ment, the copper content was generally higher in the plant tissue where copper was applied to the plots. Experiment 11 Figure 2 shows an over-all view of the onions, carrots, and potatoes in Experiment 11 during the 1954 season. Sudan grass. Data in Table 11 show that Sudan grass growing on plots receiving an application of either of the copper carriers significantly outyieldeod those plots on which no copper was applied. The yield from plots receiving 12.5 pounds of copper in the form of the oxide was significantly higher than yields obtained from plots receiving the other copper treatments. According to Table 11, applied c0pper consistently increased the content of copper in the Sudan grass tissue. Furthermore, the 23 FIGURE 2 AN OVER-ALL VIEW OF THE PLOTS IN EXPERIMENT II SHOWING THE ONIONS, CARROTS, AND POTATOES DURING THE 1954 SEASON 24 \...I...-I v.r.r.. : .nwlahf‘.. . . 5:93.}.an 19.5% I.I..o..........\.. ...h9. Hutd‘lgnkv . I.“ - .. if. -. 4...... dil'ffiH TABLE II 25 THE EFFECT OF TWO RATES OF APPLICATION OF COPPER OXIDE AND COPPER SULFATE ON THE YIELD AND COPPER CONTENT OF SUDAN GRASS AND ONIONS Sudan G ras s Onion Bulbs Pounds 1953 1954 Copper Of Carrier Copper Tons Bags per per Percent per Percent Ac re Ac re Copper Ac re Coppe r No copper . ........ 0.0 2.9 0.00083 871 0.00025 Copper sulfate ..... 3.125 4.8 0.00088 1235 0.00030 Copper oxide ....... 3.125 5.0 0.00108 1141 0.00037 Copper sulfate ...... 12.5 5.1 0.00098 1159 0.00040 Copper oxide ....... 12.5 6.4 0.00125 1212 0.00052 L.S.D. (5 percent level) 0.7 159 26 c0pper content of the plant tissue was higher when grown on plots where c0pper oxide was used than where the carrier was the sulfate. Onions. The data in Table 11 show that a significant yield response was obtained from plots receiving an application of either of the copper carriers over those receiving no copper. There was, however, no significant difference in yields between plots fertilized with the two carriers or between plots treated at the two rates of application. The copper content of the onion bulbs was highest in each case where the copper oxide was applied. The copper content of the plant tissue where copper sulfate was applied was also higher than in the tissue from the check plots. Carrots. As shown by the data in Table III, carrots in 1953 responded significantly in yield to the application of copper oxide at both rates. There was no significant difference in yield resulting from the two rates. Copper sulfate did not increase the yield re- gardless of the rate used. The percent of copper in the carrot roots was less where copper was applied than it was in the roots from the check plots in 1953. At the low rate of application, the type of carrier did not TABLE III 27 THE EFFECT OF TWO RATES OF APPLICATION OF COPPER OXIDE AND COPPER SULFATE ON THE YIELD AND COPPER CONTENT OF CARROTS Pounds 0f Tons er Acre P r nt C 2 Copper Copper p e ce opper Carrier Applied 1953 1954 1953 1954 An— nually No copper ..... 0.0 15.3 26.2 0.00034 0.00044 Copper sulfate 3.125 16.1 34.0 0.00028 0.00055 Copper oxide 3.125 19.5 32.4 0.00028 0.00055 Copper sulfate 12.500 15.8 32.3 0.00021 0.00058 Copper oxide 12.500 20.5 33.0 0.00025 0.00058 L.S.D. (5 per— cent level) ..... 3.3 4.3 Average of four replications. Bulked replicate sample, two determinations of oven-dry tissue. 28 affect the copper content of the carrot roots. At the higher rate, carrots from plots treated with the oxide had a higher copper con- tent than those obtained from plots on which copper sulfate was used. In Table III the data show that a significant response in yield for the 1954 carrot crop was obtained from both c0pper carriers. However, there was no difference in yield reSponse from the two rates of copper applied. Table III shows also that in 1954 the copper content of the oven-dry plant tissue was the same for both carriers at equal rates of applied copper, and the copper content of the tissue was higher where copper was applied to the plots than where no copper was used. Seasonal difference in yield of any one crop may be greater than the effect of treatments in any one year as indicated by the yields of carrots. Potatoes. Table IV shows that potatoes in 1953 did not re- spond to an application of copper regardless of the carrier or the rate of copper application. In fact, the tubers from the check plots were higher in content of copper than were those from the plots where copper was applied. 29 TABLE IV THE EFFECT OF TWO RATES OF APPLICATION OF COPPER OXIDE AND COPPER SULFATE ON THE YIELD AND COPPER CONTENT OF POTATOES Pounds of Bushels .Percent Copper Co er Co er er Acre1 m the Oven-Dry pp. PP. P Potato Tubers2 Car rie r Applied - 1 1 4 A” 953 95 1953 1954 nually No copper ..... 0.0 474 398 0.00030 0.00057 Copper sulfate . . 3.125 435 453 0.00018 0.00028 Copper oxide . . . 3.125 386 422 0.00023 0.00039 Copper sulfate . . 12.50 417 432 0.00024 0.00046 Copper oxide . . . 12.50 439 421 0.00026 0.00048 L.S.D. (5 percent level) ........ N.S. 28 Average of four replications. Bulked replicates, two determinations. 30 While there was a trend toward higher potato yields from copper applications in the 1954 season, the only significant differ- ence caused by the carriers occurred at the low rate of copper ap- plication. This difference was in favor of the copper sulfate. When comparing copper treatments to the no-copper treatment, copper sulfate gave a significant re3ponse in the yield of potatoes at both rates of application. Where c0pper oxide was applied, the plots produced higher yields than did the check plots; but the difference was not significant. Deficiency_symptoms. In Experiment 11 the only visual symp- toms of copper deficiency appeared on the check plots of the Sudan grass in 1953 and on the onions in 1954. The onions shown in Figure 3 illustrate the characteristic color difference caused by copper. The need for copper by the other crops was indicated by the greater yields of crops produced on the copper-treated plots. No visual symptoms of copper deficiency were observed on the potato or carrot plants from the check plots. Copper removal. Table V was prepared by converting the yields of crops to the oven-dry basis and then computing pounds of copper per acre removed by the crop. FIGURE 3 COLOR DIFFERENCE CAUSED BY COPPER (left, normal onions; right, copper—deficient onions) 31 COPPER REMOVED PER ACRE BY SUDAN GRASS, POTATOES, ONIONS, AND CARROTS TABLE V 32 P d 0:? 5 Pounds of Copper in Crop Copper Copper S Carrier Applied udan Onions Pota- Car- Grass 1 1 An- 1954 toes rots 1953 nually No copper ......... 0.0 0.041 0.012 0.019 0.020 Copper sulfate ...... 3.125 0.073 0.020 0.011 0.029 C0pper oxide ....... 3.125 0.093 0.023 0.014 0.030 Copper sulfate ...... 12.500 0.086 0.025 0.016 0.028 Copper oxide ....... 12.500 0.138 0.035 0.017 0.031 Average of two years. 33 In every case where equivalent rates of copper were applied as oxide or sulfate, more copper was removed by the crops from the copper oxide treated plots (see Table V). This may be asso- ciated with solubility and subsequent tie~up of copper from the sul- fate, whereas copper oxide being more slowly dissolved may be available to plants over a longer period in the growing season. If the amount of copper removed per acre by the crop is considered as an index to the availability of copper to plants, the relatively water-insoluble Calumet Brown Copper Oxide was more available to the plants than the soluble copper sulfate. The amount of copper removed per acre in crops is very small. For example, the 1953 Sudan grass cr0p from the no-copper plots contained 0.041 pound of copper and from the plots treated with 25 pounds per acre of copper oxide, 0.138 pound. This repre- sents an increase in yield of 112 percent. However, the difference in copper removed per acre in the two treatments amounted to 0.097 pound, which is equivalent to 44 grams of elemental copper. When the same treatments as above are compared for the 1954 onion crop, the increase was 39 and 200 percent for the yield and capper content, respectively. However, the difference in copper removed in these two treatments amounted to 0.023 pound, or 10 grams of elemental 34 copper per acre. Thus, an additional 10 grams of copper removed per acre in the onion crop made a difference in yield of 39 percent, or 341 bags. Experiment III For the placement of the minor element carriers in 1953 and minor element and insecticide mixtures in 1954 in contact with the seed of onions, the drill (Figure 4) developed by Davis, Cumings, Hulbert, and Hansen (9) was used. Onions. The data in Table VI show that 25 or 50 pounds per acre of copper sulfate applied in contact with the seed significantly decreased the stand and yield of onions. When c0pper sulfate was applied in combination with any other minor element carrier there was also a significant decrease in the stand and yield of the crop. Likewise, manganese sulfate applied in contact with the seed at the rate of 200 pounds per acre significantly decreased the stand and yield, whereas the 100 pound rate of application was not detrimental. Copper oxide at 12.5 and 25 pounds, manganese sulfate at 100 pounds, NU M at 60 and 120 pounds, and zinc sulfate and NU Z at 15 pound rates of application per acre each applied alone and placed 35 FIGURE 4 FERTILIZER PLACEMENT DRILL 36 37 TABLE VI THE EFFECT OF RATES OF APPLICATION OF SEVERAL MINOR ELEMENT CARRIERS PLACED IN CONTACT WITH THE SEED ON THE STAND AND YIELD OF ONIONS (1953) Car rie r s and Pounds Applied pe r Acre Plants Cop- er Treat- per Man- Bags I:10 ments Sul- COP- ga- Zinc per Feet per NU NU Acre fate Ox— nese Sul- of , Sul- fate Row ide fate A 0 0 0 0 0 0 595 327 B 25 0 0 0 0 0 331 118 C 0 12.5 0 0 0 0 673 405 D 0 0 100 0 0 0 574 379 E 0 0 0 60 0 0 694 427 F 0 0 0 0 15 0 715 427 G 0 0 0 0 0 15 725 438 H 50 0 0 0 0 0 74 53 I 0 25 0 0 0 0 655 415 J 0 0 200 O 0 O 387 208 K o o o ‘ 120 o o 669 393 L 0 0 50 0 0 0 669 401 M 0 0 0 30 0 0 720 384 N 25 0 100 0 0 0 204 78 O 0 12.5 100 0 0 0 532 331 P 25 0 0 60 0 0 375 166 Q 0 12.5 0 60 0 0 641 375 R 25 0 100 0 15 0 183 86 S 0 12.5 0 60 0 15 604 397 L .S.D. (5% level) 154 103 38 in contact with the seed did not significantly affect the stand or yield of onions. Each time a carrier significantly decreased the stand of onions, a significant depression in the yield occurred. Copper sulfate at 25 and 50 pounds per acre were the only treatments that significantly decreased the stand and yield of onions in the 1954 crop as Shown in Table VII. However, in 1954 the toxic effect was reduced somewhat probably by the dilution effect of the diluent applied with the copper sulfate. ‘m‘fl-Ow.“ n.‘ - ‘w .9 . 'r 39 TABLE VII THE EFFECT OF RATES OF APPLICATION OF SEVERAL MINOR ELEMENT CARRIERS PLACED IN CONTACT WITH THE SEED ON THE STAND AND YIELD OF ONIONSl (1954) _ Carriers and Pounds Applied per Acre Plants per Treat- Cop- Cop- Man- , Bags 40 ga- Zinc per ments per per U NU 2 Feet nese Sul- . Acre Sul- Ox— M Z of , Sul— fate fate ide Row fate A3 o o o o o o 1006 365 B4 o o o o o o 1005 346 C5 o o o o o o 905 293 D 25 0 0 0 0 0 585 176 E 0 12.5 0 0 0 0 955 360 F 0 0 100 0 0 0 898 279 G 0 0 0 60 0 0 877 298 H 0 0 0 0 15 0 803 299 I 0 0 0 0 0 15 854 356 J 0 12.5 100 0 0 0 830 288 K 0 12.5 0 60 0 0 940 383 L 0 12.5 100 0 15 0 795 357 M 0 12.5 0 60 0 15 903 339 N 0 12.5 0 60 15 0 923 375 O 0 0 50 0 0 0 900 386 P 0 0 0 30 0 0 868 347 Q 0 12.5 50 0 0 0 858 324 R O 12.5 0 30 0 0 871 344 S 0 0 150 0 0 0 889 356 T 0 0 0 75 0 0 887 351 U 0 25 0 0 0 0 852 302 V 50 0 0 0 0 0 593 199 W 0 12.5 100 O 0 15 742 279 X 0 0 0 0 0 O 905 293__ L.S.D.(5%1evel) 172 102 1 All plots received a basic application of 1,000 pounds of 5410- 20 per acre placed 2 inches below the seed at planting time. Average of 3 replications and expressed as 50-1b. bags/ac re. No material placed in contact with the seed. 4 Attaclay, the insecticide carrier, applied at 50 lb. per acre. 5 Treatments C through X received 50 pounds of attaclay (an insecticide carrier) per acre containing 1.5 percent duldrin and 3 percent thiram mixed with the minor element carriers and applied in contact with the seed. SUMMAR Y A. comparative study of Calumet Brown Copper Oxide and copper sulfate was made to measure their effects on the yield and copper content of Sudan grass, potatoes, carrots, and onions. Two rates of ammonium molybdate were applied to measure yield response of Sudan grass to this element. A. further study was conducted with six minor element carriers placed in contact with the seed to meas— ure their effects on the stand and yield of onions. The six carriers used were: (1) copper sulfate (25 percent copper); (2) Calumet Brown Copper Oxide (50 percent copper); (3) manganese sulfate (23.65 per- cent manganese); (4) NU M (a neutral compound containing 41 percent manganese); (5) zinc sulfate (36 percent zinc); and (6) NU Z (a neu- tral compound containing 55 percent zinc). The findings of the studies were as follows: 1. Good precision in results from the determination of cop- per in plant tissue was not obtained when dry ashing in an electric muffle furnace was practiced. However, excellent precision between duplicate results was obtained by a nitric-perchloric-sulphuric acid digestion method of ashing. 40 41 2. Sudan grass did not respond significantly in yield to the application of ammonium molybdate. 3. In the first year of copper application to a virgin, ,copper- deficient muck, the response of Sudan grass and carrots to 25 pounds of copper oxide per acre was significantly greater than when copper sulfate was applied at the same rate of total copper. 4. Very little difference in yield occurred between the two rates of copper applied or the two copper carriers in the second year, although in general, the crops responded significantly to cop- per applied. 5. The COpper content of the plant tissue was usually higher from the copper oxide treated plots than from plots where equivalent quantities of COpper were applied as copper sulfate. 6. More copper per acre was removed in the crops from- the COpper oxide treated plots than from plots receiving the same amount of copper in the form of copper sulfate. This indicates that the relatively water-insoluble copper oxide may be more avail- able to plants than the highly soluble copper sulfate. 7. The 3-1/8 pounds per acre rate of copper applied for two successive years met the needs of the crops since no response in yield was obtained from heavier applications. However, the copper 42 content of plant tissue was Slightly higher at the higher rate of cop- per application. 8. Copper sulfate at 25 or 50 pounds per acre placed in contact with the seed significantly decreased the stand and yield of onions, whereas copper oxide applied at equivalent rates of copper in contact with the seed of onions was not detrimental. 9. Manganese sulfate when applied at 200 pounds per acre in contact with the seed of onions significantly depressed the stand and yield; however, the 100 pounds per acre rate was not harmful. 10. Other minor element carriers applied either in combina— tion or alone and which had no harmful effects when placed in con- tact with the seed of onions were: (1) NU M at 60 and 120 pounds per acre; (2) NU Z at 15 pounds per acre; and (3) zinc sulfate at 15 pounds per acre. LITERATURE CITED Armiger, W. H., W. H. Hill, C. Pinkerton, H. W. Lakin, and W. 0. Robinson. Composition and fertilizer value of spent phosPhate catalysts from the petroleum industry. "Solid phosphoric acid catalysts" and copper pyrophos- phate catalysts. Journ. Amer. Soc. Agron. 39: 318-326, 1947. Association of Official Agricultural Chemists. Official and Tentative Methods of Analysis, 6th edition, Banta Press, Menosha, Wis., pp. 121-122, 1945. Berger, K. C., and E. Truog. Re3ponse of sweet corn to fer— tilization with copper and zinc. Soil Sci. Soc. Amer. Proc. 13: 372-373, 1948. Brown, J. C. The influence of copper compounds applied to muck soil on the yield, growth pattern and composition of certain crops. ”Ph.D. thesis," Michigan State Col- lege. 1949, PP. 96. Brown, J. C., and P. M. Harmer. The influence of copper compounds on the yield, growth pattern, and composi- tion of spring wheat and corn grown on organic soil. Soil Sci. Soc. Amer. Proc. 15: 284-291, 1950. Butler, L. 1., and H. 0. Allen. Copper and cobalt in plants. Jour. Assoc. Official Agr'l. Chem. 25: 567-573, 1942. Callan, R., and J. A. R. Henderson. A new reagent for the colorimetric determination of minute amounts of copper. Analyst 54: 650-653, 1929. Coulsen, E. J. Report on copper. Jour. Assoc. Official Agr‘l. Chem. 20: 178-488, 1938. 43 10. 11. 12. 13. 14. 15. 16. 17. 18. 44 Davis, J. F., G. A. Cumings, W. C. Hulbert, and C. Ms Hansen. A fertilizer placement drill designed for organic soil investigation. Michigan Agr. Expt. Sta. Quart. Bul. 32: 250-255, 1949. Elvehjem, C. A. The biological significance of copper and its relation to iron metabolism. Phys. Review 15: 471-507, 1935. Harmer, P. H. Studies on the effect of copper sulfate applied to organic soil on the yield and quality of several crops. Soil Sci. Soc. Amer. Proc. 10: 284-294, 1945. Harris, H. C. Copper deficiency in relation to the nutrition of oats. Soil Sci. Soc. Amer. Proc. 12: 278-281, 1947. Hill, M. F., and O. C. Bryan. The nutritive relation of copper on deficient soil types of Florida. Jour. Amer. Soc. Agron. 29: 809-814, 1937. Johnson, K. E. E. The effect of methods and rates of applica- tion of two copper carriers on the yield and COpper con- tent of spinach and sudan grass grown in the green- house on two organic soils, "Unpublished M.S. thesis," Michigan State College, 1952, pp. 39. Kretschmer, A. E., Jr., and W. T. Forsee, Jr. The use and effectiveness of various copper bearing materials for application to everglades organic soils. Soil Sci. Soc. Amer. Proc. 18: No. 4, 471-474, 1954. Lipman. C. B., and G. McKinney. Proof of the essential nature of copper for higher green plants. Plant Physiol. 6: 593-599. 1931. Lucas, R. E. The effects of copper applied to organic soil, alone and in association with manganese and zinc, on composition of creps and reactions in the soil. "Ph.D. thesis," Michigan State College, 1949, pp. 83. Rehling, C. J., and E. Truog. Milorganite as a source of minor elements for plants. Jour. Amer. Soc. Agron. 32: 894- 906, 1940. 19. 20. 21. 22. 23. 45 Ruby, D. C. Relationship between soil acidity and copper defi- ciency in muck soils. I'Unpublished M.S. thesis," Michigan State College, 1951, pp. 41. Sommer, A. L. Copper as an essential element for plant growth. Plant physiol. 6: 339-345, 1931. __1___ Copper and plant growth. Soil Sci. 60: 71-79, 1945. Steenbjerg, F., and E. Boken. Copper contents and copper de- ficiency in Danish soil types. Plant and Soil 2: 195— 201, 1950. van Eck, W. A. The effect of copper carriers on crop produc- tion in organic soils. "Unpublished M.S. thesis," Mich- igan State College, 1954, pp. 66. ('1‘. "I .Q . RU? ' 37 'l Ua‘fii (55:5 0‘.” it. WI 3 MICHIGAN STATE UNIVERSITY LIBRARIES I Illllllll I ill 3 129313103 8791