II II I I II I' ,1 ., III: In . ; I" II ,I III “I "I ’ II ; I I I III III II III III III III III II III I I \f— K" w I” -_'(o_-‘ IxJM I .030 I! RELATZQNEHW SETWEEN SOIL AfiiDlT‘i’ AM} (TOWER DEEIQENfl‘I" {N MUCK 561$ ‘fixesEs ("at if“ D-iigre-e 65 55%. 5. MSCHIGAN STATE LCLQ“ {Emaid Q Ruby Wfi‘ on" Thieietoeertlfgtlntthe thesis entitled . " . , "The Relationship Between Soil Acidity and Copper Deficiency in Muck Soils" preeented by de Ce Ruby 5 has been accepted towards fulfillment 3) of the requirements for 14.3st degree in 501]. Science ; Fwd/.1, .“‘. ,1 I ‘~ ABSTRACT Ruby, Donald C. Relationship between soil acidity and copper deficiency in muck soils. The purpose of this study was two fold; A. An investigation of the effect of the following factors on the capper content of muck soil and of Henry spring. wheat grown on this muck: (1) The addition of cepper ‘ sulfate as a soil amendment and as a spray. (2) The add- ition of capper oxide as a soil amendment. (3) Th6 appli- cation of ashed cepper deficient muck soil with and without acidification with hydrochloric acid. B. To investigate the relationship of soil reaction on the copper content of virgin muck soils and on the leaves of pOplar trees (Populus tremulgides) growing on these muck soils. A OOpper deficient muck soil from the Michigan State College Muck Experimental Farm was treated in the green- house as follows: A. Check - no cepper added. B. COpper sulfate (Cusoh°5H20) added at the rate of 100 pounds per acre. cement he --.-‘ .- ’» Ruby, Donald C. C. Eguivalent amount of COpper supplied in the form of ( 4.8%) cOpper oxide as applied in Treatment B. D. Ash obtained from burning one foot of muck surface soil, untreated. E. Same as Treatment D except the ash was acidified with hydrochloric acid. F. Plants sprayed with 0.186 percent solution of copper sulfate. G. Ash obtained from burning six inches of surface muck, untreated. H. Same as Treatment D except an attempt was made to remove the copper by suspending iron nails in a hydrochloric acid solution of the ash. All treatments included 3-9-18 fertilizer at the rate of 3000 pounds per acre. The cepper content of both the soil and tissue was determined by the Carbamate Method. The following data were obtained: 1. Chloride ion toxicity was believed to have caused stunted growth of Henry spring wheat when used to acidify alkaline ash material. 2. The results indicated very little relationship between pH and copper content of the soil as compared to the cOpper content of the wheat plant. 3. Wheat plants showing no copper deficiency symptoms were grown on both alkaline and acid muck. ii Ruby, Donald C. h. Leaching with distilled water reduced the copper con- tent of a muck to which ash acidified with hydrochloric acid was added. I 5. Organic soil and acidified ashed muck with MnSO °5H20 h added at the rate of 100 pounds per acre and subsequently leached, produced plants with no copper deficiency symptoms. Muck samples varying in degree of acidity and pOplar leaves were obtained from seven areas located in Allegan, Clinton, Ingham, Jackson, Lepeer and Sanilac Counties. The pH and copper content of the muck and the copper con- tent of the leaves were determined in the laboratory. The following results were obtained: 1. As muck becomes more acid the copper content of the soil tends to decrease. 2. From the data it appeared that pH and cepper content of the soil at six of the locations had little effect on the copper content of the peplar leaves. The exception to the general trend occurred in the Baker Farm sample. In this case a possible iron - copper relationship may be present. 111 RELATIONSHIP BETWEEN SOIL ACIDITY AND COPPER DEFICIENCY IN MUCK SOILS By» Donald 0. Ruby_ 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 1951 iv ACKNOWLEDGMENT The author expresses his appreciation for the assist- ance, counseling and guidance given him by the following staff members: Dr. J.F. Davis, Dr. K. Lawton and Dr. L.M. Turk, Department of Soil Science. Dr. E.J. Benne and associates, Department of Agri- cultural Chemistry. Dr. H.L. WDmechel and wr. D. Harvey, Department of Mechanical Engineering. The writer acknowledges the many helpful suggestions and cOOperation of his fellow graduate students and to Mr. R.D. Bailey of the Buildings and Utilities Division for his timely assistance in the fabrication of special ashing equipment used in the experiment. IX. XI. XII. XIII. XIV. TABLE OF CONTENTS ABSTRACT TITLE PAGE ACKNOWLEDGMENT TABLE OF CONTENTS FIGURES TABLES INTRODUCTION REVIEW OF LITERATURE A. Early work in the United States with cOpper B. Copper application under Michigan conditions 0. Copper in laterite soil EXPERIMENTAL PROCEDURE A. Treatments B. Ashing process C. Copper determination- Carbamate Method D. Standard curve EXPERIMENT I. The Effect of Soil Amendments on the First CrOp of Henry Spring Wheat EXPERIMENT II. The Effect of Soil Amendments on the Second Crop of Henry Spring Wheat EXPERIMENT III. Relationship of the Soil Reaction and the Copper Content of Virgin Muck with the Copper Content of POplar Leaves GENERAL DISCUSSION GENERAL SUMMARY BIBLIOGRAPHY vi Page iv vi vii viii N l--'\O\l\l our-bow I--' bud H O\ 21 28 33 38 LC Eigure Page ——a- I. Standard curve for COpper determination. 1A II. The effect of the addition of copper sulfate and of ashed muck acidified with hydrochloric acid on growth of Henry spring wheat in the greenhouse (first crOp). 18 III. The effect of the addition of copper sulfate and ashed muck,untreated,on the growth of Henry spring wheat in the greenhouse (first crap). 19 IV. The effect of the application of cepper sulfate on a cepper-deficient muck and ashed muck acidified with hydrochloric acid with subseouent leaching on the growth of Henry spring wheat in the greenhouse (second crop). 23 V. The effect of the application, on a c0pper- deficient muck, of cOpper sulfate and ashed muck acidified with hydrochloric acid in which iron nails were suspended in an acidified solution of the ash to remove cepper from solution, with subsequent leaching, on the growth of Henry spring wheat in the greenhouse (second crop). 24 VI. The effect of the application of copper sulfate and ashed muck,untreated, on a c0pper—deficient muck, on the growth of Henry spring wheat in.the greenhouse (second crop). 25 VII. The relationship of soil reaction to the percent copper of virgin muck soil and pOplar leaves. 31 vii TABLES Table 1. Dilutions of cOpper sulfate solution used in the construction of the standard curve. 2. The effect of various treatments on the cepper content of a c0pper-deficient muck and on Henry spring wheat grown on these different treated jars under greenhouse condition. 3. The effect of pH and copper content of virgin mucks on the percent of c0pper in poplar leaves (Populus tremuloideg). viii 17 30 RELATIONSHIP BETWEEN SOIL ACIDITY AND COPPER DEFICIENCY IN MUCK SOILS INTRODUCTION In'Michigan, there is an estimated 5,000,000 acres of organic soils representing approximately one acre in eight for the state. Several different cr0ps (8)1 are grown on muck2 with celery, onions and mint the most important of the special crops group. However, practically any crOp may be grown on muck provided proper soil management pract- ices and adapted varities are employed. One of the import- ant factors encountered in the successful production of crops is the correction of copper deficiency that occurs in many of the muck areas. Results from previous work (7,8, 12,22) have indicated that copper deficiency is associated with soil reaction, and in Michigan for a number of years the soil pH has been used as a basis for making practical field recommendations. A pH of 6.5 has been recognized as the upper limit at which cOpper deficiency is likely to occur. However, it is also recognized (9) that a diff- 1-The numbers in parentheses refer to literature cited. 2-The term "muck" is used to include both muck and peat. erential crop requirement exists and copper is recommended for some crops (head lettuce, spinach and wheat) even with a pH above this arbitrary value of 6.5. In view of the fact that there is an apparent assoc- iation between copper deficiency and soil reaction, this investigation was instituted to study the factors that might affect this relationship. If cepper deficiency and soil reaction are closely correlated then the following hypotheses might be formulated: 1. Acid mucks deficient in cepper because of the increased solubility of c0pper compounds in an acid medium and subsequent leaching would remove the copper from the medium. 2. The mineral soil areas surrounding the muck form- ation were low in copper and the resultant drainage water collecting in the muck area was low in this element and thus the resultant copper content of the plants forming the muck area was very low. 3. The availability of copper is closely correlated with soil reaction. A. COpper deficiency in a muck soil is corrected by burning due to the resultant concentration of copper. A consideration of some of these factors is included in this work. REVIEW OF LITERATURE Early work with copper in the United States was re- corded in Dade County, Florida in 1927. Reprecht (18) showed that tomatoes responded to soil applications of cepper sulfate. Felix (6) found that failure of lettuce and onions on unproductive mucks in New York in later stages of growth was corrected by the addition of cepper sulfate. Results of soils investigations of Florida (2) on several species of field crOps and truck crops showed a marked plant response to copper. Plots on which cepper was applied produced in most cases apparently normal growth of plants in constrast with an absolute and early failure on those areas from which the treatment was withheld. Results from Allison's (1) experiments with the application of COpper sulfate on peanuts were positive. Hunter (10) using coPper treatments on several grasses revealed that an increase in both fresh weight and dry weight per acre resulted as compared to the fresh and dry weights obtained from the untreated plots. Data from the Canal Point Breeding Station (3) of Florida showed that sugar cane responded to an application of thirty pounds of c0pper sulfate per acre when applied to raw sawgrass peat. Harmer (7) found that under Michigan conditions that some species of plants responded to COpper sulfate and that COpper deficiency was associated with soil reaction. Results from investigations at that time indicated that fifty pounds of copper sulfate per acre was sufficient to correct cepper deficiency for most creps which are copper responsive. Spinach and lettuce crops, however, required an application of one-hundred pounds per acre for complete control of the trouble. Additional application of twenty- five pounds per acre for succeeding crepe gave good re- sponses. Later he found through several observations that crOp response on muck in Michigan differ (9). As a result of this a classification of crOps based on the response to copper was set up. Liming an unproductive acid peat soil did not always result in a favorable growth for corn according to the data of Willie and Piland (22). An unfavorable condition de- ve10ped by liming is associated with excessive absorption of iron and lodgement of iron in the nodes of the plant. This situation was remedied by the addition of copper sulfate. Further investigation (23) by these men indicated that cOpper serves as a catalyst in oxidation-reduction reactions. Orth and others (1?) pointed out that cOpper is nec- essary for the formation of chlorophyll in the leaves of orange trees. It was also suggested that cattle grazing in Florida on certain pastures developed nutritional anemia or "salt sickness” and this trouble may be corrected by an application of iron or copper to the grazing area. Harmer's (8) results suggested a direct effect of copper on the formation or function of the chloroPhyll in plants. According to Lucas (13) the increase in copper con- tent in plants was as much as threefold, although only a very small fraction of the copper contained in a normal application of coPper is recovered by the crop. Experimental results presented by Jamison (ll) sug- gested that since copper is largely retained in slowly soluble or slowly replaceable form, it appears that a small continuous supply in the soil may be more important than the amount of copper that is easily replaceable. An or- dinary soil application of cOpper would doubtless be toxic were it not for its fixation. Brown's (5) observations indicated that copper is necessary for normal metabolic activity of plant processes. Lucas (12) showed that under Michigan conditions the copper requirement in plants was related to the copper con- tent and pH of the soil and possibly to the nutrient balance within the plant. A paper by Lundblad (it) reports that the ratio of cepper in the soil and in vegetation does not seem to be essentially different in calcareous soils and acid soils. No relationship between pH and cepper deficiency in organic soils has been reported in Sweden (15). No significant corrlation as reported by McVickar (16) could be observed between potash, phosphorus, magnesium and calcium content or the calcium-magnesium ratio of the white oak leaves and corresponding values for the A horizon of the soil upon which the trees grew. Vermaat and van der Bie (20) investigated certain lateritic soils and found the c0pper released through weathering was mainly adsorbed in the exchangeable ionic form. When the concentration became too great in these soils growing rubber trees, the 00pper intake was increased and the stability of the latex was adversely affected, an important factor in shipping. Soils found to have a higher amount of organic matter had a lower concentration of 00pper in the ionic form. It was concluded that cepper was fixed in the molecule of the organic matter. Thus by increasing the organic matter by mulching ionic copper was incorporated in a molecule of circulating organic matter. EXPERIMENTAL PROCEDURE Greenhouse experiment The following eight treatments replicated four times were set up in two-gallon glazed jars in the greenhouse and two crops of Henry spring wheat were grown on these treated jars. H. Check-no cOpper added. Copper sulfate (CuSOh’SHZO) at the rate of 100 pounds per acre. Equivalent amount of copper supplied in the form of (6h.8%) cOpper oxide as applied in Treatment B. Ash obtained from burning one foot of muck sur- face soil, untreated. Same as Treatment D except the ash was acidified with hydrochloric acid. Plants sprayed with 0.186 percent solution of copper sulfate. Ash obtained from burning six inched of surface muck, untreated. Same as Treatment D except an attempt was made to remove the copper by suspending iron nails in a hydrochloric acid solution of the ash. Note- all jars contained a copper-deficient muck. A virgin copper-deficient muck was obtained from the Michigan State College Muck Experimental Farm. This area had been recently cultivated but never fertilized. An analysis of a sample from the same general area was report- ed by Brown (5) as follows: Organic matter, 86.0%; total nitrogen, 3.3%; Potassium, 0.21%; phosphorus, 0.12%; calcium, 2.5%; magnesium, 0.27%; iron, 1.3%; and copper, 0.0011%. The muck was partially dried and screened through a one centimeter square mesh screen to remove root and wood fragments. A uniform application of 3000 pounds of 3-9-18 fertilizer per acre was applied to all Jars. Chemically pure grades of ammonium nitrate, potassium di-phosphate and potassium chloride were used in the preparation of this fertilizer. In order to eliminate any possible induced manganese deficiency resulting from the alkaline ash add- ition, manganese sulfate (MnSOL°5H20) at the rate of 100 pounds per acre was applied to all Jars in Treatments D and G. On completion of first experiment when analysis reveal- ed that iron nails did not remove copper from the acid medium in Treatment H, manganese sulfate at the rate of 100 pounds per acre was added to this set of Jars. The required ash, fertilizer and copper materials for each Jar were thoroughly mixed with 5&00 grams of muck and lightly packed in the Jars. The manganese sulfate was applied as a top dressing. Twenty seeds of Henry spring wheat were planted on two different dates June 19 and August 28 in each jar and theemerging plants were later thinned to 6-10 plants per jar. Ashing process- Muck weighing 6000 grams and of uni- form moisture content (this was assumed to be equivalent to one foot of surface muck from an area of the same dimensions) was placed in a polished steel box of 11 gauge metal with the dimensions of 6"x20" and 8" high. The box was covered with an asbestos plate perforated with several holes one quarter inch in diameter. Three boxes of this type and size were prepared together with two smaller boxes 5"x12" and 3" high. The muck samples were then placed in a gas furnace located in the Forging Building and temperature held be- tween 600-650 degrees centigrade for six hours. This furn- ace could accommodate three samples at one time. The covers were removed after the first three hours of the ash- ing process. In this process the muck was reduced to approximately one-third of its original volume. The samples were then transferred to an electric muffle furnace and held at about 650 degrees centigrade (a dull red color of the box) and the ashing process continued for another 8 to 20 hours. 10 The weight of the ash from the above samples varied between 300 to 350 grams. A number of difficulties were encountered in this ashing process: l~ The steel boxes deteriorated rapidly as was evidenced by scaling and resultant slight contamination of the ash with the iron. 2- Corroded contact points on the muffle furnace. 3- One hour of manual operation of the gas furnace was necessary before it would operate automat- ically. Acidifying the ash- Approximately 300 milliliters of concentrated hydrochloric acid was required to reduce the pH of the ashed muck to 6.36, the pH of the original muck sample. Increments of the acid were added over a 48 hour period. Removal of cOpper from solution- One dozen iron nails were suspended in an acidified sample of the ash. This pro- cedure was based on the electromotive-series theory that copper in the solution would be replaced by the iron and the copper deposited as a coating on the nails. Harvest- The wheat was harvested at the time the seeds were in the milk stage of maturity. 11 Laboratory Experiment Determination of copper in plants- A composite sample of the straw and grain from the four replications of each treatment was dried at 65 degrees centigrade. The plant material was then ground in a Wiley Mill and this prepared plant tissue used in the determination of c0pper. Between 3 to 5 gram samples of plant material representing each treatment was oven-dried at 105 degrees centigrade, weighed and ashed. The ash was treated gently with con- centrated hydrochloric acid and then.brought to a boil for several minutes. The solution was then transferred to a 200 milliliter volumetric flask and brought up to volume for each determination. Aliquots were taken from each flask for copper analysis and the c0pper content determined by the Carbamate Method. Determination of copper in muck soil- The same pro- cedure was used for the determination of the copper content of the muck as was used for the plant material. A thor- oughly mixed sample of 3 to 8 grams of muck soil was oven- dried at 105 degrees centigrade, weighed and ashed. The ash was then heated gently with concentrated hydrochloric acid and bought to a boil for several minutes. The solu- tion was placed in a 200 milliliter volumetric flask and 12 brought up to volume. COpper was determined in aliquots of solution from the flask. Standard Curve- In using the Carbamate Method a standard curve was set up to chart the cepper concentra- tions. The standard copper solution was made up by dis- solving O.7587 grams of anhydrous copper sulfate (Merchs reagent quality 99.688% pure) in distilled water and diluted to one liter. One hundred milliliters of the above solu- tion was then diluted to one liter thus resulting in a con- centration of 0.03 milligram of cOpper per milliliter. The latter stock solution was used in the construction of the standard curve (Fig.1) based on the concentrations re- ported in Table l. Cepper was determined colorimetrically using a Lumetron photo-electric colorimeter model AOO-A and a lantern blue glass filter of A20 mu. Some of the difficulties encountered in the Carbamate Method of copper determination are as follows: 1- Too small amounts of copper in the sample resulted in transmission values close to the blank transmission value. Thus, the error was greatly increased in the determination of the copper content. This difficulty of high transmission read- ings was partially overcome by taking larger aliquots for copper analysis and (or) larger samples of material. 2- Elemental iron proved to be an interfering ion in some soils. When iron interference was realized smaller aliquots 13 and (or) smaller samples of muck were used in the determin- ation. 3- At first the filtering process was used to separate the residue from the liquid portion of the sample Table l- Dilutions of cepper sulfate solution used in the construction of the standard curve. - .g..._- U-’ ~—--.1. -m‘. "I. .o a for cOpper analysis. Mg/ml copper I Ml. : Ml. distilled I cOpper water 'f0.030 I 10.0 00 I 0.020 I 6.7 : 3.3 1 0.015 I 5.0 : 5.0 0.010 I 3.3 : 6.7 I 0.006 I 2.0 : 8.0 0.005 I 1.7 : 8.3 0.001 I 0.3 : 9.7 0.000 I 0.0 : 10.0 -.:‘-..A.—. ntm---_-- “ a.- -I-..- run—-- -:..'-.n,-- -- -y..- —- --— -. 0 -.1.- Lantern blue glass filter of A20 mu. percent transmission nH/u—m—w n—r- ---.- -— -1m—mm I“ 7 12 18 31 A7 54 86 100 - 9‘ r...s no- as; -n a -.....-.<—:.— .n—- .-,_.,,. A This procedure was found to be very time-consuming even when used under a vacuum. A check on the experiment using solutions with and without residue in the bottom of the volumetric flask was found to give pract- ically the same results. dispensed with. The filtration process was then 6w:- .u:o:o: :00 came... 0 3.... 2.3.5.... .I. 3 PERCENT OPPER 15 Muck samples and leaf samples from p0plar trees (Regulus tremulpigeg) were obtained from seven areas located in the state with pH ranging from b.35 to 7.88. The copper contents of the soil and leaf samples were obtained accord- ing to the procedure outlined in Experiment III. EXPERIMENT I The Effect of Soil Amendments on the First Crop of Henry Spring Wheat The experiment was set up as previously described in Experimental Procedure. Henry spring wheat was planted June 19, 1950 at the rate of approximately 20 seeds per jar. In general all plants germinated in normal length of time except in jars of Treatments E and H. Emergence of plants was retarded approximately one week by the hydrochloric acid treatment. When the plants reached a height of h to 6 inches they were thinned to stand leaving 6 to 10 plants per Jar. Plants of Treatment F had two applications of cOpper solution sprayed on its leaves. These applications were made on July 21 and 28, 1950. A good stand of wheat was obtained in all jars except in those of Treatments E and H. In these pots the plants remained stunted throughout the course of the experiment. The untreated muck soil contained 0.0030 percent cOpper which was higher than was anticipated. Soil from a near-by area analyzed 0.0016 percent c0pper as shown by data obtained from experiment III. The relatively high c0pper l7 .pooaanomno ma use: mesa amdawano uoono on mouo coooem and; cocoa pnoaoeoanb .uoouuo 0Nmm.40mq2 on one on has museum awaken one menace opoaou pod 0H0 uaaoan .Hoanopoa human who venues nod one nomqooum .ooapooaanouoo nommoo non Hofiuopea pecan awoode.uo2n4 .unoaauomwe no magnum one mnaodawon one he come one; doaaonaanopoc we owe neqmoo «on oeaqaomnm I .'|l\(l '10 .‘A. .II. 1 It. a..- - useenonaeem..00.m-. mmH00.0 . - 4:4n00.0 egomw seodoauae . w . uuommoo dsqu unease Hasnoz. 0m.0 5400.0 5000.0 ensues eons use; noeooo madman Hausa knob 0m.0 0000.0 4a-: no Hesoaon unsouao , . .20ee 00000 00H0fi0fl0a_ . _ negate Heeuoz 0m.0 «400.0 ~A00.0 onenexaa anon asset euzonm Hassoz n . 00.5 , 4400.0 mH00.0. 0000a 0000mm wam_ _ n , _. aseonm Heeeoz 0m.0 M~m00.0 wm0mH0.0 commune wowH.0. assonw Haanoz x H 00.0 ,mm00.0. m0ea0.0 ooaseaom sooooow mousse Hausa w40.x. W ”0400.0 M 0 ma00.0. mom apes eoaeaeaosl madman Hanan numb m n 0m.0 _ 4000.0. 41:: Moss comma pooh moon 1 . w . . aesoem Haenoz .00.w W ,m000.0 m x HH00.0 oeansana petal essoem Haaeoz . 00.0 a “0000.0. ma00.0 woes 00am0 0000 000, 0 n _ . U . Boson» Haeuoz “00.0 m .0mn0.0 “ . «H00.0w hem.400 000 0a nooooo. apeonw Heeeoz _ , 0m.0 .0ma0.0 _ ma00.0 M0 seeoas peoaepae0m” _ _ n . . epsouw aesuoz .0m.0 A r300.0 n ma00.0. 4 once ~00 I assoum Haenoz * . 0m.0 l wH000.0 . ma00.0. 0mmm. omeo «00000 00am _ . . . assonw naeeoz 000.0 1 .0m00.0 “ mn00.0 . aces. assoem Huseoz . i 0m.0 . .Hm00.0 em00.0 eeoaoauoeueooooo I.y - - .-us .znnues.-nar. It» ; .1 _ -.. na:-, - . -a , I . Macho m mono mono n mono_ mono mono 00m 00 . 00m;:er:.20m0, 00m 00H uMHoEom a. A 0-2211- -1 Haom . anode nanosecona _ _ . . ;. mn-: - uwnmoo paoouom_ mommoo unconom_ fiI ‘ .10 t.l . . I‘( III. I I) . ll () (villll'li - .. I... y‘, )I l'll' t- .mv.ooo«paoaoo omsonaoonw nouns anon cocoon» oneneuuau omega do qsonw poems weanmu madam no one Mose unoaoauocnuoqmoo o no pseudoo noneoo one do mpnoauoonu mooanop no poouue one um canoe Fig.II. The effect of the addition of copper sulfate and of ashed muck acidified with hydrochloric acid on the growth of Henry spring wheat in the greenhouse (first crop). 0- no copper; l- 100 pounds copper sulfste per acre; 3- ashed muck acidified with hydrochloric acid. 19 ‘ ‘ . n -- '/ , -.—.\ - ,‘n -‘ 0 _.. D- - . Fig. III. The effect of the addition of copper sulfate and of ashed muck (untreated) on the growth of Henry spring wheat in the greenhouse (first crop). 0- no copper; l- 100 pounds c0pper sulfate per acre; 2- ashed muck (untreated). 20 content may explain the reason for normal growth of plants in the jars receiving no copper (Fig. II). Also, in Fig. II theapparent toxicity of the hydrochloric acid treatment may be observed. As shown by the condition of plants in Fig. III the addition of one foot of ashed muck without hydrochloric acid treatment did not interfere with the normal growth of wheat. Plants sprayed with 0.186 percent cOpper solution did not appear to be copper deficient. In general the pH of the soil and copper content of the soil had little if any effect on the copper content of the plants. Although the plant material of the check jar indicated a high cepper content this may have been due to outside contamination unknown to the writer. Wheat plants grown on these same jars under conditions of Experiment II actually showed cOpper deficiency symptoms. 21 EXPERIMENT II The Effect of Soil Amendments on the Second Cr0p of Henry Spring Wheat 3 The purpose of the experiment was to study any pos- sible differential effect that might occur with a second crop in view of the fact the general area from which this muck was obtained had previously been shown to be a cOpper-defic- ient muck. After the harvest of the first crop of Henry spring wheat the muck soil was screened through a one centimeter square screen to remove most of the roots of that cr0p and the soil again replaced in each individual 2-gallon jar from which it had been removed. Each jar of soil was screened separately to avoid contamination. The muck from jars of Treatments E and H was transferred to h-gallon jars and leached with t.5 gallons of distilled water in order to remove the excess chloride ions that may have caused toxicity to the plants. This procedure required approxi- mately one week to complete. Fertilizer (3-9-18) was applied at the rate of 2000 pounds per acre to each 2-gallon jar. Chemically pure materials were used in preparing the fertilizer as in Exper ime m: I . 22 Another Treatment, J, was added to the experiment in which a known copper-deficient muck was used as a check. The soil of these jars was fertilized with 3000 pounds per acre of 3-9-18 fertilizer. Twenty seeds of Henry spring wheat were sown on August 28, 1950 in each jar. No apparent delay in emergence of plants in any treatment was noted. When the plants reached a height of A to 6 inches they were thinned to stand leaving from 6 to 10 plants per jar. Two applications of 0.186 percent copper sulfate solution were applied to plants in Treatment F on October 20 and 27, 1950. Plants were harvested on December 15, 1950 at the time heads were emerging from the boot. As indicated by the data in Table 2 there is little apparent relationship between pH and percent cOpper in the plant or percent cepper in the soil to the percent copper in the plant. Plants from Treatment H were very low in copper content as compared to plants from the other treat— ments. The fact that an addition of manganese sulfate to jars of Treatment H apparently corrected a condition ob- serVed in the plants growing in jars to which acidified ashed muck was added might indicate that a manganese deficiency was induced by the subsequent leaching process. 23 Fig. IV. The effect of the application of cepper sulfate on a capper-deficient much and ashed muck_acidified with hydro- chloric acid with subsequent leaching on the growth of Henry spring wheat in the greenhouse (second crop). 0- no cOpper; l- 100 pounds cepper sulfate per acre; 2- ashed muck acidified with hydrochloric acid and leached. 2h Fig. V: "The efEESE”6r EH; application, on a capper deficient muck, of capper sulfate and ashed muck acidified with hydro- chloric acid inwhich iron nails were suspended in an acidi- fied solution of the ash to remove copper from solution, with subsequent leaching, on the growth of Henry spring wheat in the greenhouse (second crop). 0- no capper; l- 100 pounds cOpper sulfate per acre; 3- ashsd muck acidified with hydrochloric acid, attempt cOpper removal by iron nails, added manganese sulfate 100 pounds per acre and leached Fig. VI. The effect of the application of copper sulfate and ashed muck (untreated), on a copper-deficient muck, on the growth of Henry spring wheat in the greenhouse (second crop). 0- no cepper; l- 100 pounds copper sulfate per acre; h- ashed muck (untreated). 26 As shown in Fig. IV plant growth was still adversely affected. This poor growth seems to be due to manganese deficiency rather than chloride toxicity. Huck soil in jars shown in Fig. V seem to produce plants with normal growth under similar conditions as above plants except manganese sulfate was added. The plants produced on the muck receiving no COpper were cepper deficient in Experiment II. A logical expla- nation as to the reason for the failure of the plants in the first crOp of wheat produced on the same jars to develop cepper deficiency symptoms is difficult to establish other than that the original sample of muck contained sufficient copper for one crop of wheat but not for the second crop. According to the data in Table 2 leaching with distil- led water reduced the copper content of the muck from 0.0064 percent to 0.0046 percent. The data also showed that copper was not removed as a result of suspending iron nails in the acidified ash solution. Plants showing copper deficiency symptoms were found only in jars from the check treatment. The appearance of plants (Fig. VI) showed that the addition of ashed muck did not adversely affect the growth of wheat under the conditions encountered in this experiment. Manganese sulfate was applied to correct any possible induced 1.0 new 27 manganese deficiency resulting from the increased alkalinity from the ash application. It was assumed that copper defici- ency did not deve10p because of the copper contained in the ash and not because of any soil reaction relationship. Plants sprayed with an 0.186 percent copper solution did not deve10p cepper deficiency symptoms. However, the relative- ly high copper content of plants from this treatment as re- ported in Table 2 were in part due to the failure to wash the excess cepper remaining on the leaves from the spray applica- tion. Cepper added in the form of 00pper oxide was apparently as effective as an equivalent amount of copper sulfate in cor- recting copper deficiency in wheat plants. No logical ex- planation for the comparative contents of copper in muck treated with eduivalent amounts of copper oxide and copper sulfate can be advanced. an application of one-half the amount of ash was as effective in preventing copper deficiency in wheat as was double this amount. Plants growing on the Known cOpper deficient muck (Treatment J) showed typical copper deficiency symptoms such as die-back of the tips, loss of turgor and tissue necrosis. The symptoms developed by plants from Treatment A were similar but not as pronounced. 28 EXPERIMENT III Relationship of the Soil Reaction and the Copper Content of Virgin Muck with the Copper Content of Poplar Leaves The purpose of this experiment was to compare the copper content of some virgin mucks, the pH of these mucks and the amount of cOpper in a standard plant growing on these soils. POplar trees (Pppglug tggmglpiges) were selected because they can be faind growing over a wide range of conditions. Samples of seven muck areas were obrained within a 150 mile radius of Michigan State College in Southern Michigna located in Ingham, Jackson, Clinton, Sanilac, Lapeer and Allegan Counties. The samples were taken at the depths of 0-6 inches and 2h inches. Approximately 300 leaves from each of two poplar trees growing on these areas were obtained at the time of the sampling of soil. The muck samples were taken between the two trees from which the leaf samples were obtained on July 25 and 28, 1950. The pH of the muck soils was determined with a glass electrode. The leaves and soil were analyzed for COpper by the Carbamate Method. The poplar leaves were dried in an oven at 65 degrees centigrade to facilitate grinding. Then the leaves were ground in a Wiley hill and placed in steppered glass jars until analyzed. The ground leaf material was then thorough— ly mixed and oven-dried at 105 degrees centigrade. Samples of 3 to 5 grams were weighed out and then ashed at approxi— mately 650 degrees centigrade. The ash was taken up in con- centrated hydrochloric acid and brought to a boil for several minutes then transferred to a 200 milliliter flask and brought up to volume. The data presented in Table 3 showed that with the ex— ception of the Baker Farm data the pH of the soil had very little relationship to the copper content of peplar tree leaves. The pH of the muck soils on the Lynn Farm were 4.27 and 4.70 for the 0-6 inch and 24 inch depths respectively and resultant values for copper content were very low. The COpper content of poplar tree leaves growing on this soil was not essentially lower than the copper contents of peplar leaves on other muck areas investigated. The pH of the muck samples from the Walling Farm for the 0-6 inch and 24 inch depths was 7.83 and 7.53 respectively. 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