SUME ENTERRELATIONS 0F FERTILIZERS. CARBOHYDRATE PRODUCTION AND YIELD OF FIELD BEANS THESIS FOR THE DEGREE OF M. s. C. W. Lauritzen 1932 ‘(Lv‘qgguvmdé €37. MIRA!" A.“ an mum at. SOME INTERRELATIONS OF FERTILIZERS, CARBOHYDRATE PRODUCTION AND YIELD OF FIELD BEANS by .1 y . ‘\ C'.‘ w.’ LAURITZEN rTmL‘“ A THESIS PRESENTED TO THE FACULTY OF THE MICHIGAN STATE comer: OF AGRICULTURE AND APPLIED SCIENCE IN PARTIAL memmm or THE REQUImmMS FOR THE DEGREE OF MASTER OF SCIENCE East Lansing 1 9 3 2 THFQ‘S ACKNOWLEDGMENT The writer wishes to express his appreciation to Dr. C. H. Spurway for his invaluable guidance and interest shown during the course of this investigations Thanks is due to Dr. C. E. Millar and Dr. J. W. Grist for the critical reading of the manuscript, also to many others who have contributed to this research by their suggestions and criti- cisms. 0 (j ‘ “a; -r . 1 N __ l I -1- IHTRODUCTIOK The yield obtained from any economic plant is dependent upon the many factors which influence the growth of the crop under con- sideration. Commercial fertilizer applications may be one of these factors, depending on whether there exists a soil deficiency of the elements supplied. If applications of a commercial fertilizer produce a marked effect upon the yield, the other factors remaining unchanged, it is assumed that the plant nutrient elements supplied have or have not been limiting factors in the production of the crop, depending on the yield changes produced by the fertilizer. Carbohydrate production is undoubtedly a necessary process in the growth of plants. Since fertilizer elements affect plant growth the question is raised as to what part these fertilizer elements play in the synthesis of the various carbohydrates found in plants and what relation exists between carbohydrate production in growing plants and the yields obtained. In order to determine if such relationships exist in the case of bean plants, and also to determine if the rate of production of carbohydrates at dif— ferent times of the day is influenced by the fertilizer elements supplied, bean plant parts were analyzed for carbohydrates, nitrogen, 'and dny weight in plant samples taken at four-hour intervals from check plots and from plots receiving applications of various fertilizer elements. .A considerable amount of research on carbohydrate production in plants has been carried out using several kinds of plants. -2- These researches have been.mainly attempts to determine the order of synthesis of the different plant carbohydrates and certain relations they bear to one another; as the variation in the amounts present in the plants with the change of tempera- ture, intensity of sunlight, and consequently the time of day. Other investigations made along this line have had for their purpose the determination of the functions of nutrient elements in the synthesis of carbohydrates in plants. No one, to our knowledge, has attempted to correlate carbo- hydrate production during periods of growth with.the eventual yield; or has used field beans in researches dealing with carbo- hydrate production. Beans are a valuable food crop high in carbohydrate content, and they are extremely sensitive to environ- mental factors which cause uncertainty in their profitable pro- duction. Because of the importance of this crop from the stand- point of both production and consumption, studies of this nature are desirable. -3- HISTORICAL Noble (9), working with crops that store large amounts of carbohydrates such as sugar beets and mangolds, found that a sufficient supply of potassium was necessary for the normal production of carbohydrates in the plants. Stocklasa (16) showed that green plants did not have the power of synthesis in the absence of potassium. Reed (11) found evidence in the plants he studied that potassium was necessary for the formation of starch, but gave no evidence that phosphorus was necessary for starch.formation. He showed, however, that a deficiency of phosphorus seriously interfered with the transformation of starch into water- soluble carbohydrates. Weiszmann (18) made the same observation as Stocklasa. Smith and Butler (1h) could not find evidence to support Stockflasa and Weiszmann. Schuster (12) pointed out that the results obtained from an application of potassium to a crap depended on the amount of available potassium in the soil as well as other soil properties, and stressed the importance of considering the soil type as a significant factor in planning an investigation of this nature. Janssen and Bartholomew (h) found that tomato plants grown in mediums low in potassium.showed an increase in the per cent of dry matter, nitrogen, and sugars. They also reported (5) -h- that the percentages of carbohydrates and potassium in the plants fluctuated greathy and concluded that a.high.per cent of sugars and starch was not necessarily associated with a high per cent of potassium.in the plant. They found however a fairly good correlation between the total weight of sugars and starch per plant and the potassium content. MacGillivary (6) showed that in the case of tomatoes a deficiency of phosphorus resulted in an increase in the per cent of total nitrogen, this increase being mostly water soluble forms of nitrogen, excluding nitrates, but caused a decided increase in the per cent of sugars. Stark (15) noted that when potassium was applied to soy- beans, in addition to limestone and organic matter, a decrease in the per cent of protein resulted; but limestone and organic matter alone gave an increase in the per cent of protein. Davis, Daish, and Sawyer (2), after making a study of the formation and translocation of sugars in mangolds, reported that the proportion of saccharose in the leaf tissue followed the temperature curve closely during the daytime; and that the pro- portion of hexoses increased faster than the temperature, and in such a.manner that the curve showing the ratio of hexoses to saccharose in practically parallel to the temperature curve. Davis and Sawyer (3) found in their study of potato plants that saccharose was the principal sugar in the plant when the -5- tubers were being formed, and that the amount increased from sunrise until 2 P.M. following approximately the rise in tempera- ture. The starch content of the plants reached its maximum point two hours before sunset. From that time on the amount of starch decreased till there was very little present at 2 A.M. {ason and Maskell (7) studied the transport of carbohydrates in the cotton plant. They found a significant lag in the diurnal variations in the bark over those in the leaves and that the ‘ variation of the total sugars was due largely to the variation in the concentration of sucrose. The rate of transport of carbohydrates from the bark to the boll in cotton plants was found to correlate with the variation in the sucrose gradient from the bark to the boll, the rate being about four times as great in the day as in the night. Clements (1), working with potato, soybean, and sunflower plants, showed by his graphs that the period of maximum carbohydrate production was from noon until about 9 P.M. The increase in starch content was especially noticeable during this period. A distinct correlation was found between favorable environmental conditions and the formation and translocation of carbohydrates in the plant. Cole (19) very recently working with the potato plant, found irregular variation in the sugar content of plants grown on plots receiving various fertilizer treatments but no differences were found which could be attributed to fertilizer application. He expresses the Opinion that under field conditions other factors such as intensity of light, moisture, and humitity enter in to -6- cause variations in the sugar content of the potato plant to a greater extent than does the relative supply of nutrient elements present. 1 rd.. 3.12511 - 23618.. - . - 2.1200,- -‘ FIGURE I 1010 101A 1013 o - 16 . 0 POUO 1020 102‘ 1023 o - 16 ' 0 3.0. fl 43? 4’ 1030 103A 1033 § {‘3 0 - 16 - 8 P.U. .8 O 3 101m 10111 10113 03 «‘3 a O " 16 b 8 3000 ,o 3 A 1050 1051 1053 O E a a 0 - 16 - 8 P.U. 8 h - o 4- 0 3.00 M g 1060 1061 1063 3 m M - 16 «3 PM. 1070 107A 1073 I" - 16 "' 8 30°. KEY 1030 1031 10813 P.U. Plowed under 3.0. Broadcast 1" - 16 u 8 Pen. 1090 1091 1093 )4 - 16 ‘L 8 3.6. v/ °sqtoou 1/4111 oo -7- EXPERIMENTAL PROCEDURE The bean plant samples used in this research and the yield results were obtained from c00perative fertility plots located in the vicinity of Ithaca, Michigan, on the soil type known as Brockston loam. Figure 1 shows the plan of the fertility field, the kinds and amounts of fertilizers applied, the methods of applying the fertilizers, and the location of the check and treated plots from.which the plant samples were taken for analysis and from which the yields were obtained. Judging from the yields of crops produced previously on this land, the field was above the general average in fertility for the locality. The bean yields reported in this thesis, however, are much.be10w the average for the Brookston soil type in this State, due probably to the deficiency of rainfall during the growing season of this year (1930). During the year pre- ceding the establishment of the bean fertility plots this field was cr0pped to corn and was plowed early in the Spring of 1930 for beans. The bean fertility plots were laid out and the fertilizers applied in such a way as to make possible comparisons of yields, and carbohydrate production of beans on soil receiving no treat- ment; superphosphate alone; superphosphate and.muriate of potash; and nitrate of soda, superphosphate, and muriate of potash. Three methods of fertilizer application were used; namely, (1) broadcast before plowing, (2) broadcast on the plowed and -g- worked field before seeding, (3) superphosphate and muriate of potash plowed under and nitrate of soda broadcast on the plowed and worked field before seeding. ,All treatments were applied at the rate of four hundred pounds per acre, except on plots 108 (A.and B) and 109 (A and B), which were ferti- lized at the rate of two hundred pounds per acre. The ferti- lizers were mixed according to the following formulas: 0-16-0, 0-16-8, and M-lG-S. Then they were carefully broadcast by hand on the plots, having first been diluted by mixing them thoroughly with about twice their volumes of moist soil to prevent any possible blowing and to insure a more uniform distribution over the plots. The beans were planted June 12, 1930, and harvested September 23, 1930. The plots used to obtain plants for carbohydrate analysis were 102A, lOHA, 107A, and the check strip as designated (Figure 1). Plots 1023, thB, and 1073 were harvested and the yields determined for comparison with the carbohydrate production of the plants on the.A portion of the corresponding plots. The yields on plots 1010 to 1070 inclusive, were averaged and this average value taken for comparison with the carbohydrate pro- duction on the check strip. The day preceding sampling was warm.and.part1y cloudy. During the period of sampling the weather was clear and warm, except for the period between four o'clock P.M. and eight o'clock P.M., when it was partly cloudy; -9- The bean plants at the time of sampling were green having pods somewhat more mature than string beans when taken for table use. Some variation was observed in the maturity of the plants as is always the case; but on the average, it might be said that the plants were of such maturity that the pods contained beans from one-half to full size. The plant sglples were taken every four hours during a period of twenty- four hours. The first samples were taken at midnight August 13, 1930, and the last samples at midnight August 1h, 1930; thus, the first and last samples were taken at the same time on two different days. The samples were collected by taking plants promiscuously over the plots, the several samples being gathered at one time and taken as quickly as possible to the field laboratory close by where the equipment for preserving the samples was located. The leaves, stems, and pods of the plant were separated.by hand. Fifty grams of each part which.previously had been cut into pieces one to two inches in length were placed in pint Jars containing approximately one gram of Ca003 to neutralize any acids present which would tend to hydrolyze the sugars; the presence of an excess of 0a003 being unimportant as its dissociation was such that the pH was not raised enough to cause the breaking down of the sugar molecule which occurs at higher concentrations of hydroxyl ions. The Jars were then nearly filled with boiling 95% ethyl alcohol; the caps turned on loosely and the Jars set in --10-- a water bath hot enough to keep the alcohol at the boiling point in order to prevent all enzymic action and partially extract the sugars from the plant tissues. At the end of one hour the jars were removed from the water bath and enough more hot alcohol added to cover the plant tissue, in case some had been lost due to evaporation. The Jar caps were then tightened down and the jars placed in storage until wanted for analysis. -11- ANALYTICAL METHODS Analysis of Bean Tissue Preliminary treatment of the sample: The field sample preserved in alcohol was filtered into a 1000 cc. volumetric flaSk and.washed with 80% alcohol until the filtrate was about 950 cc. This volume was made up to 1000 cc. and was called the ”extract”. The residue was placed in a 200 cc. weighed beaker and dried in an oven at 65° 0. for M8 hours, cooled in a desicator and weighed. The aliquots for analysis were calculated on the basis of this weight. Treatment of the Extract The alcohol from 300 cc. of the alcoholic extract was distilled off under reduced pressure in a classion flask below 65 0. When about 50 cc. remained the residue was washed from the classion flask into a 100 cc. volumetric flask, with warm water, cooled, made up to volume and transferred to a 100 cc. centrifuge tube. About 0.2 grams of anhydrous lead subacetate (Horn's dry lead) was added, mixed well, and the samples centri- fuged for five minutes. The clear liquid was transferred to another centrifuge tube and de-leading by adding about 0.5 grams of sodium oxalate, mixing and centrifuging five minutes. The clear solution was decanted off into a beaker and 75 cc. of it taken and placed in a 200 cc. volumetric flash; ‘Water was added -12.. to make the contents of the flask up to volume. The Determination of Dextrose Fifty cc. of the solution were placed in a 300 cc. Erlenmeyer flask and 50 cc. of Fehling's solution were added—-25 cc. of EA” (69.28 grams CuSQu.5déO dissolved in water and diluted to 1 liter) and 25 cc. of ”B" (3M6 grams Rochelle salts and 100 grams NaOH dissolved in water and diluted to 1 liter). The flask was placed in a water bath at 80°C. for 30 minutes (10). At the end of this time the flasks were removed and cooled to about #000. The determination was continued using the Schaffer and Hartmann.method (l3). Twenty-five cc. of an iodide-iodate solution (60 grams KI, 5.2 grams K103, and a few drops of concentrated NaOH dissolved and diluted to 1 liter with.water) was added with shaking. Next, 15 cc. of 8N HéSOu were added and the flask shaken until the Cu20 was dissolved. .Then 20 cc. of a saturated solution of K2020h were added and the contents of the flask titrated at once with standard‘ NaZSZO3 sol- ution (2M.82 grams NaZSZOjfifléo dissolved in water, a few drops of concentrated NaOH added and the volume made to 1000 cc. with water). The burette reading was the number of cc. of 178128203 used to oxidize the excess of iodine present which was the amount liber- .- Jour. Assoc. Official Agri. Chem. 12: 169 (1929). -13- ated by the acid less the amount used in oxidizing the Cu20. The amount of iodine which reacted with the Cu20 was obtained by subtracting the number of co. Na25203 taken to titrate the sample from the number of cc. Na25203 taken to titrate the blank determination. The blank consisted of 50 cc. of water run in duplicate, with each set of determinations which re- ceived exactly the same treatment as the samples and gave the total amount of iodine liberated. 0112501,} 21: 01112 211825203; 1: NaQSuO6I- 2Na 1 Since one equivalent of Cu required one equivalent of I for oxidation, the difference in the titration of the blank and the sample in cc. multiplied by the Cu equivalent in.milligrams per cc. of the NagSng solution equalled the Cu reduced by sugars and was converted to dextrose by referring to the lunson and Walker tables (8). The dextrose in the determination times 17.777 (dilution factor) equals dextrose in total sample. When the titration is referred to in cc. of Na25203 it should be understood that it is the co. difference in the titrae tion of the sample and its corresponding blank determination. The Determination of Total Sugars Fifty cc. of the same solution used for the determination of dextrose in the extract was placed in a 300 cc. Erlenmeyer flask. Five grams of citric acid crystals were added and the contents of the flask boiled quietly for ten.minutes, then cooled and neutral- ized with neon using phenolphtalein as an indicator. The deter- -1h- mination is continued as for dextrose except 20 Cc. of 8N sésoh instead of 15 cc. are used. The cc. Na28203 times Cu equivalent of Na25203 solution equals Cu reduced in the deter- mination. Dextrose equivalent of the Cu times 17.777 (dilution - factor) equals total sugar in sample in terms of dextrose. The total sugar minus the dextrose times 0.95 equals hydrolyzable sugars or sucrose. The Determination of Dextrine The dry residue was ground in a Wiley mill and a three- tenths aliquot weighed out in a 150 cc. beaker, 75 cc. of water and a few cc. of toluene were added and the sample allowed to stand over night. The solution was filtered off into a 250 cc. volumetric flask and the residue washed with warm water until the flask contained about 150 cc. of the filtrate. To this filtrate was added sufficient concentrated HéSOh to make it 25% of HéSOu. The mixture was shaken and let stand over night, then made up to volume and a 50 cc. aliquot withdrawn and neutralized with concentrated NaOH using phenolphthalein as an indicator. The contents were allowed to cool and the determination for dextrose made at once. The number of co. Na25203 times Cu equivalent of Na25203 solution equals Cu in determination. The dextrose equivalent timOI 16.666 (dilution factor) equals dextrine in sample reported as dextrose. The Determination of Starch The residue from the dextrine determination was transferred to the original 150 cc. beaker using about 50 cc. of water and -15- the starch in it determined by the Take-Diastase Method (17). In this method the beaker containing the residue was heated for #0 minutes on a steam bath with stirring. After cooling, enough TakaeDiastase to digest the starch (0.1 gram Taka-Diastase to 0.3 grams of starch) and 3 cc. of toluene were added. The samples were incubated in a constant temperature oven at 3830 for an hours, with stirring at intervals, more toluene being added to replace that lost by evaporation. After digestion was com» pleted the mixture was heated to boiling on a steam bath and boiled for 15 minutes; then the residue was filtered into a 250 cc. volumetric flask and washed until nearly that volume is ob- tained. Water was added to complete the volume and a 50 cc. ali- quot taken for the determination of dextrose. The total starch in 50 cc. equals 51 f 52(17) 2. 31 . E? Total reducing power of the digested starch. The starch in 50 cc. is multiplied by 16.666 (dilution factor) to obtain starch in original sample of bean tissue. Determination of Dry Weight 100 cc. of the extract representing a one-tenth aliquot was placed in a weighed 200 cc. beaker (tall form) and the alcohol evaporated off at a low heat on an electric hot plate. A.one-tenth aliquot of the corresponding dry ground tissue was then added to the same Beaker and the contents of the beaker -16- dried at 105°C. for 2% hours, cooled in a desiccator and weighed. From.these weighings the per cent dry weights were calculated. Determination of NitrOgen The loose, dry residue from the dry weight determination above was transferred from the beaker to an 800 cc. Kjeldahl flask to which was added 15-18 grams of KZSOM and 1 gram of Cusou. The residue dried on the beaker was washed off by first warming with 15 cc. of concentrated HZSON and then dislodged by means of a rubber policeman and another 15 cc. of cold concen- trated Hésou and finally washed out with a few cc. of water. From this point the procedure was the same as that employed in the determination of nitrogen by the Kjeldahl method. DISCUSSION The results of this research showed some evidence of a relation between the percentages of sugars, starch, and dry weight in the bean plant parts and the time of day. .Also, that the average amount of starch found in the plant parts was related to the fertilizer treatment and the yield of beans. Contrary to the results obtained in 1915 by Davis, Daish, and Sawyer (2), who worked with.mangolds, it will be seen by observing graphs 1 and N that the per cent of sugars in the bean leaves at different times of the day did not follow the daily temperature changes, but, if anything, tended to be low during periods of high temperature and high during periods of low tempera~ ture. If the stems and pods alone were considered, there was a noticeably low content of sugars during the period from 8 A,r. to 8 9.1.1. (graphs 2, 3, 5, 6, 19, 20). In their work with potatoes, Davis and Sawyer (3) obtained similar results for sugars in the leaves of mangolds and reported in addition a general increase in the per cent of starch from sunrise until two hours before sunset. This work on beans showed some indication of a maximum starch content in the leaves at about 8 P.M. (graphs 10 and Ml); but a period of low starch content was shown during the middle of the day. The stems and pods of the beans did not show a maximum starch content as in the case of leaves at 8 P.M., but rather at some time between 12 K. and N Alh., and a minimum point at 12fi. -13- (graphs 10, ll, 12, 22, ”1, and MB). The variations in per cent dry weight followed the temperature changes although with some degree of lag; consequently it seemed that a higher per cent of water in the plants was associated with a higher per cent of sugar and starch. During the entire sampling period of 2% hours, the per cent of dextrine and nitrogen was fairly constant in all the plant parts and no variation due to treatments was evident. It will be noticed that a collective average of the per cent constituents in the leaves.stems, and pods was made. However, it was known that this average did not represent the exact per cent of the constituents in the whole plant, but was taken for the purpose of comparison, as it tended to produce uniformity in the results and conclusions, thus causing the curves of the per cent constituents with the time of day to become smoother and the results of the several treatments to more nearly parallel each other. Considering the average per cent of the several constituents determined in leaves, stems, and pods separately, some very notice- able relationships become evident, as to the relative amounts of the constituents in the different plant parts. Dextrose was found to be present in the greatest amounts in the stems and the least amounts in the leaves (graphs 1, 2, 3, 25, and 37). Sucrose, on the other hand, although contained to the least amount in the leaves, existed in greater amounts in the pods than in the stems (graphs h, 5, 6, 26, and 38). The greatest per cent of starch was -19- in the pods with the least amount in the stems. Carbohydrates, which were the sum of the constituents named above and dextrine, which was present in nearly the same amount in each.plant part, increased in amounts from leaves to stems to pods. The per cent dry weight followed the same order in the plant parts as did the per cent carbohydrates. Thus, it will be seen that the bean plant tissue which contained the least water had the largest per cent of carbohydrates, and yet, as was previously shown, the water and carbohydrate content of the plant parts fluctuated tagether. Nitrogen was present in about the same percentages in the leaves and pods with considerably less in the stems. The above relationships were discovered from data gathered primarily to determine the effects of fertilizers on the per cent of certain plant constituents at different times of the day, but there seemed to be no apparent relationship shown in this respect, and no attempt will be made here to explain these data on the basis of plant processes. The fertilizer treatments did not appear to have any distin- guishable effect on the per cent sugars in the plant except perhaps the 0-16-8 (graph 25). In this case there was evidence that the per cent dextrose was depressed. When the daily average was con- sidered the per cent starch in the bean plant parts showed a rather definite correlation with the fertilizer application. The bean plants grown on the plot receiving an application of super- phosphate and muriate of potash tOgether showed an increase in -20= the per cent of starch over those grown on the plots receiving superphosphate alone and those grown on the plots receiving no treatment (graphs 29, 35, MN, M5). The addition of nitrate of soda to superphosphate and muriate of potash further increased the per cent of starch in the leaves and stems, but depressed the per cent in the pods. The depressing effect of nitrogen in the fertilizer treatment on the per cent starch in the bean pods might be accounted for in that the plants on this plot may have been a little less mature and starch was not being accumu- lated there to the extent it would have been if the plants had been more mature. The plants on this plot seemed a little greener at the time the samples were taken than the plants on the other plots. Considering the fertilizer nutrient elements separately, po- tassium in the fertilizer apparently increased the per cent of starch in the bean plant parts (11); or at least a combination of phosphorus and potassium increased the per cent starch over no treatment or phosphorus alone. The increase in the yield of beans paralleled the increase in the per cent starch and carbo- hydrates, as a whole, in the bean plant parts (graphs uh, M5, and #6) (tables u, 7, and 8). Therefore, the yield seemed to be related to the per cent starch and also to the total carbohydrates in the bean plant parts at this stage of maturity. Both the per cent carbohydrates at this time, and the yield, were related to the fertilizers applied. -21- SUMMARY A study was made of carbohydrate production, per cent nitrOgen, and the dry weight of the bean plant relative to the time of day and fertilizer application. A.relation was found between carbohydrate production and the time of day, but no apparent relation was evident between fertilizer application and percentage constituents at various times of the day. It was found, however, that there was a marked difference in the per cent composition in the leaves, stems, and pods of the plant, and that the relation varies with the constituent under consideration. It appeared that potassium did increase the percentage of starch and total carbohydrates in the bean.plant parts and that this increase was reinforced by the addition of nitrogen or perhaps it should be said that phosphorus and potassium together gave an increase in carbohydrates which was further increased by the addition of nitrogen. This increase in carbohydrate production at this period of growth was paralleled by a corresponding increase in yield. Per cent dextrose in bean plant parts from check and fertilized field plots calculated on the dry weight basil 0 TABLEI Average of Treatments... 1L--16--8 0-16-0 0-16-s Check Time Leaves 70272715 88868638 0 O O O O o O 0 00000010 5 3 1.1 375m9®29 0 0.00 O. 0 00100010 7.38 812‘... (wow/aw SJl-l. 0.00.0.0.0.1.0 1.01. 0.95 0.69 0.87 0 81 0.66 1.1.1 0.92 mmmfi gap...) .0....90 1.0000010 1+ 1.21. 8 ‘0“. 12 n. 1t PM. 8 P.M. 12 14. Average 12!. Stems lenne 31.66 2 8 lrhw 52 8 31.122222fl2 73 368k. mégmuaboamfll 1.12.12 95.22 2 31.01 578 .47mwh. 7h. .nw 329..1.?..22../2 Mm. mwm %r%ur=w% 333.1. 9... 2 3?. 21. 3005 1.8.1 9.1.27. 33222232 12 M. u 1.11. 8 1.3. 12 N. h P.M. 8 P.M. 12 u. Average Pods mum/kn 925% 1211.10.11. easieeao o.21.1.1.0.1.1. 37.658 756 391. 378 32 11110.0.11. 13310931 07382976 0 O. O 000 21101021 .40 936 8.43 122?. o 0 221011 1. 1.1.4. 8 1.16. 12 N. 14 P.M. 8 POM. 12 1.1. Average 12M. Average of Treatmen 17-16-8 Leaves Stems 0-16-0 0-16-8 fertilized field plots calculated on.the dry weight Check basis. TABLE 2 Per cent sucrose in bean plant parts from check and Time 90672725 Ion-8371.71. 6710115814. 36352530 0 C O C O O C O . O C ..... 22222222 33322233 62989751. 29 antennae awn 27 22221332 231W2121..J2 1.615 1.73 1135567 2637 .6su edeeeee. 3221.1«29HZ 8 33321233 d O P 02 909 623 “1. .usss 5s5.JJmm 2&122122 “331.2133 79 61. ”95055 2.7. 8M....4..%. 7.9nw81x4.%1 2221.29H9m?“ 9H3zfl239m33 mm .. m . .uu . . . . . u..n..ur . . .m A... PD. 0 MNJNvmnrnr Per cent dextrine in bean plant parts from check and fertilized field plots calculated on the dry weight basis . TABLE 3 Average of Iregtmegts 4-16-8 0-16-8 Check 0-16-0 Time Leaves 1.. 73516 56 6 96 8 7006 7 ........ 00000000 1. w. 8 1.1!. 12 N. 1‘ PO”. 8 PJI. 12 M. Average 12”. Stems ........ 00011110 M1157 39 .../580.19 90w .. ... 01000000 7.68.466 3 .4 40738 In». W. . . . . . 01000111 8 P.M. 12 M . Average 4.A.M. s 1.11. 12 N. 1. 9.3. 1211. Pods $95738 .190119 . . . . . . . 01011101 TABLE h Per cent starch in bean.plant parts from.check and fertilized field plots calculated on the dry weight b33130 . ' Time Check 0-16-0 0-16-8 11-16-8 gm Leave: 12M. 0.80 0.97 Luna 11.61 2.70 h 1.11. .99 11.80 5.36 11.90 9.76 8 1.11. .78 n.61 6.08 5.92 2.22 12 N. 11.61 5.02 3.37 3.33 .08 u 2.24. 5.3 3.79 2. 7 3.87 3.88 8 2.14. 7.8 3.69 5.30 11.80 7.16 12 u. 1.15 1.05 6.59 “($8 11.07 Ayerage ”.06 3.N2 n.80 5.92 ”.55 Stems 12 u. 0.79 1.63 14.314 11.10 2.72 h 1.11. 14.56 5.3 2.27 5.16 11.33 8 1.8. 11.76 3.02 11.32 3.25 3.91 12 n. 2.70 3.22 2.96 2. 2 2.82 n P0“. 3.8” 3060 2.10 ' 3093 3037 8 1m. .10 3.142 11.26 .78 .111 12 M. 3.22 1.56 6.02 7.97 14.69 Average 3.h2 3.11 3.75 n.56 3.71 Pods 1211. 11.95 9.81& 9.78 8.73 8.32 n 1.11. 9.52 9.95 9.01 8.69 9.29 8 1.14. 5.99 7.77 7.99 6.86 7.03 12 N. 5.60 11.97 6.87 11.117 5.118 b. 2.11. 5.61 H.87 6.90 6.12 5.88 8 P.M. 7.10 5.21» 9.62 8.79 7.69 12 M. 10.09 11.03 12.16 12.70 11.19 Average 6.98 7.67 8.83 8.05 7.88 TABLE 5 Per cent dry matter in been plant parts from cheek and fertilized field plots. nme Check 0-16-0 0-16-8 11.168 f”? ° 0| 1' Leaves 12 M. 19.80 20.23 19.911 19.90 19.l+7 n 1.11. 19.69 19.99 18.61 20.50 19. 69 8 1.11. 20.86 19.59 18. 111+ 18.78 19.112 12 N. 21.07 20.97 21. 3’4 20. 78 21. 01+ 11 2.11. 20.25 19 .911 21.16 20. 60 20.149 8 19.11. 20.10 19.13 20. 99 21.71 20.118 12 M. 19.50 19.06 21.38 21.02 20.211 1verege 20.18 19.811 20.27 20.117 20.19 Stems 12 m. 22.28 21.55 22.37 22.0 22.06 n 1.11. 21.111 21.62 20.96 22.1 21.53 8 1.11. 23. 51 20.91 22.31 20.711 , 21.87 12 n. 2.29 22.96 23.16 23.06 23.12 n ma. .09 23.611 .26 23. 67 23.67 8 17.21. 21.21 21.97 2 .91 22.111 23. 8 12 11.1.22 19.60 22.08 22.60 21. 6 Average 22. 21.75 22.72 22.38 22.33 Pods 12 M. 26.15 25.98 27.08 28.57 26.95 1 1.8. 21.93 25.21 23.76 211.25 34.31 8 1.11. 25.17 26.93 2 .113 25.03 25.39 12 n. .25 79 29.09 28.36 27.143 27.67 11 2.11. 25.145 26.140 28. 50 26.88 27.31 8 2.11. 23.1 29.911 27. 59 30.112 27.77 12 n. 27.7 25.55 28.16 27.53 27.25 1verage 25.05 27.31 27. 12 27.16 26. 66 fertilized field plots calculated on the dry weight TABLE 6 Per cent nitrogen in bean plant parts from.check and basis. Ayerage of Tr Check 0-16-0 0-16-8 4-16-8 Time Leaves 3333332 3 666 76 78 1.. #6 J?— 350/1...“ 1221212222222222 wwwmm $1“..an O O IJsz/zazzzzzzzz 4 1.11. 8 1.11. 12 NO 4 2.11. 8 PM. 12 u. Awerags 12 M. Stems 367101.. 300 1001.9 oM/OJ 22221211.. 9 9511.6 nu. 9J1hfl8 11.21.1211 71757.66 992 0.0. 2.29 21121211 5557 6 9 ”322$ 51.. 22222212 026717.071. 81.088339 0 O. .0. 12211211 0 O 0. w o o O HumuuanVmu-L 2112ppzn alhwnb.lhwao.l.1 Pods 6635 51.. 11366 mu 8 O 0 O O O 0 314. 33332 3 8 17 1 mwfimufirm 0 3huu33332 3 3.17 99 h. oranw e-le .223, .31 3-35 2.41 3.18 3 36 71.1CUIJIJ 22607122 0 as o o o o 0 33333323 12 M. 4 1.11. 8 1.11. 12 N. 4 11.11. 8 9.11. 12 M. Awerago TABLE 7 Ayerage per cent constituents in bean leaves, stems and pods from check and fertilized field plots calculated on the dry weight basis. _ Ayerage of Mme Check 0-16-0 0 16~8 4-16-8 1mm Dextrose 12 M. 2.12 2.28 1.82 1.78 2.00 4 1.11. 2.23 1.96 1.87 2.05 2.05 8 1.11. 1.54 1.87 1.4 1.88 1.68 12 n. 1. 7 0.95 1.2 1.80 1.32 4 PJI. 1.54 1.6 1.28 1.71 1.55 8 2.11. 1.37 1.53 1.22 1.34 1.37 12 u. 2.01 2.60 1.83 1.90 2.12 Average 1.73 1.84 1.53 1.78 1.72 Sucrose 12 u. 2.11, 3.14 2.74 2.36) 2.59 4 1.11. 2.36 2.71 2.74 2. 2.64 8 1.11. 2.39 2.25 2.52 2.24 2.44 12 N. 2.12 2.06 1.98 2.29 2.11 4 2.11. 2.21 2.35 1.55 1.30 1.94 8 P.1£. 2.08 1.61 2.51 2.61 2.21 12 u. 2.42 2.68 2.65 2.75 2.54 Ayerage 2.24 2.40 2.38 2.28 2.33 Dextrine 12 M. 0.58 0.55 0.54 0.55 0.56 4 1.1!. 1.33 1.05 1.08 0.80 1.06 8 1.1:. 0.73 0.96 0.90 0.89 0.87 12 11. 0,97 1.06 0.89 0.94 0.96 4 19.11. 1.02 0.96 0.88 0.99 0.96 8 1w. 1.09 1.02 0.95 1.01 1.02 12 M. 1.02 0.89 0.82 0.92 0.92 Average 0.96 0.93 0.87 0.87 0.91 CONTINUATION OF TABLE 7 Time Check 0-16-0 0-16-8 u-16-8 Average of Treatments Starch 12 u. 2.18 - 11.15 6.18 5.81 n.58 h 1.11. 6.01 6.70 5.52 6.25 6.13 8 1.11. .18 .13 . 5.28 .39 12 n. 3.30 3.110 3.30 3.111 .13 1+ 2.11. 14.95 11.09 3.82 .6n 11.38 8 2.11. 6.35 M12 6.39 8.146 6.33 12 m. n.82 11.55 8.26 9.38 6.75 Average 11.83 11.73 5.79 6.18 5.31 Nitrogen 12 u. 2.88 3.06 3.31 2.90 2.96 1+ LM. 3.27 3.23 3.31+ 3.12 3.21; 8 1.11. 3.15 3.17 3.05 3.27 3.08 ‘12 N. 2.86 2.61+ 3.22 2.99 2.93 n 19.1.1. 2.96 2.87 3.05 2.61 2.87 8 2.14. 2.98 3.08 3.00 2.86 2.98 12 M. 2&3 2.119 2.38 2.65 2.51 Average 2.9 2.93 3.05 2.91 _2.96 Per cent dry matter on green weight 12 m. 22.71; 22.59 23.13 23.51 22.83 u 1.11. 20.66 22.31 21.78 22.30 21.81; 8 1.1!. 23.18 22.118 21.73 21.52 22.2 12 n. 23.38 2u.3l+ 2.129 23.76 23.9 n ma. 23.26 23.99 21+.31 23.72 23.82 8 2.15. 21.19 23.68 211.50 2 .85 23.88 12 a. 22.93 21.1w 23.87 23.72 22.98 Average 22.52 22.97 23.37 23.31:: 23.05 TABLE 8 Average per cent carbohydrates in bean plant parts from check and fertilized field plots during a 2%- hour sampling period calculated on the dry weight basis . Check 0-16-0 0-16-8 1146-8 Leaves 7.27 7.00 7.96 8.81 Stems 9.57 9.55 3.65 11.13 Pods 12.89 13.32 1 .11 13.05 Average 9.76 9.90 10.57 11.15 TABLE 9 Yield in bushels per acre of clean dry beans. Check 0-16-0 0-16-8 h-16-8 Bushels per acrel 9.68 l 9.95 I 12.60 I 13.31 8. 90 10. 11. LITERATURE CITED Clements, H. P. Hourly variations in carbohydrate content of leaves and petioles. Bot. Gas. 89: 2114-272, 1930. Davis, I. L. Daish, A. J., and Sawyer, G. C. Studies of the formation and translocation in plants. I. The carbohydrates of the mangold leaf. Jour. Agr. Sci. 7: 255-286, 1916. Davis, I. 1., and Sawyer, G. 0. Studies of the formation and translocation er carbohydrates in plants. III. The carbohydrates of the degradation of starch in the leaf. Jour. Agr. Sci. 7: 352-38h, 1916. Janssen, G., and Bartholomew, R. P. The translocation of potassium in tomato plants and its relation to their carbohydrate and nitrogen distribution. Jour. Agr. Res. 38: uu7-u65, 1929. Janssen, G., and Bartholomew, R. P. Influence of potassium concentration in the culture medium on the production of carbohydrates in plants. Jour. Agr. Res; 1#0: 2h3-261, 1930. MacGillivary, J. B. Effect of phosphorus on the composition of the tomato plant. Jour. Agr. Res. 314: 97-129, 1927. Mason, T. G., and Haskell, E. J., Studies on the transport of carbohydrates in the cotton plant. I. A study of the diurnal variations in the carbohydrates of leaf bark and wood and of the effect of ringing. Ann. of Bot. he: 190-253. Munson, L. S., and Walker, P. E. The unification of reducing sugar methods. Jour. Amer. Chem. Soc. 28: 663-686, 190 . Noble, Uber die organishe Leistung des Kalium in der Pflanse. Landw. Versuchs Stat. 13: 321-h23 Quesumbing, F. 1., and Thomas. A. I. Conditions affecting the quantitative determination of reducing sugars by Fehling's solutions. Jour. Amer. Chem. Soc. 1&3: 1503- 1526. 1921. Reed, H. S. The value of certain nutrient elements to the plant cell. Ann. of Bot. 21: 501-518, 1907. ' 12. 13. 1h. 15. 16. 17. 18. 19. Schuster, G. L. Potash in relation to quality of crop. Jour. Amer. Soc. Agron. 19: 506-517, 1927. Shaffer, E. An, and Hartman, A. F. The codometric determination of copper and its uses in sugar analysis. Jour. Biol. Chem. M5: 365-390, 1920. Smith, T. 0., and Butler, 0. Relation of potassium in plants. Ann. of Bot. 35: 189-227, 1921. Stark, R. W. Environmental factors affecting the protein and oil content of soybeans and the iodine number of soyfiean oil. Jour. Amer. Soc. Agron. 16: 636-6H6, 192 . Stocklasa, J., and Pitra, J. Uber die Einwirkung der Kalisalze auf die Entwicklung der Gerste. Ziet. f. Landw. Versuch.im Oeeterr. P. 567. * Thomas, Walter. The colorimetric determination of carbo- hydrates in plants by the picric acid reduction method. II. The determination of starch and other "reserve" polysaccharides. Jour. Amer. Chem. Soc. U6: Part 2, 1670, 192M. Weiszmann, H. Uber den Einfluss des Kaliums auf die Entwicklung der Pflanzen und ihren morphologischen'und anatomischen Ban, bei besonderer Berucksichtigung der landwirtschaftlichen Kulturpflanzen. Ziets. f. Pflanzendungung 2: _l—79, 1923. Cole, Ralph C. The diurnal and seasonal changes in the sugar content of the sap and tissue of potato plants as affected by soil fertilization. 5011 Science 33: 3h7-362. 1932. MICHIGAN STATE COLLEGE 1 11 1 1 I <1 1 11 I 11 1 1 11417 HOU.P.79JIO.IWI. ‘IHIH an. H} OQIQOvoofi r‘ Ho..- 4 LY *fimwme WAd Hflw o It 4 a I. o ..oo...__ w .. f» 11er tree: » +0.70 m HmHHHIA H 0.. T4L*AII.I v. I e ....e ‘Ilfieieoeeeau IOII ILVIQIOIIIOIQIIO 7.61751 1 1|4 9.5 l..r..I”. 91700III IIO YVAvaveeoateoeeteoI ......v .enoeeee II Ieofi..— toHoo-I pl 0. r. _.u_.~s .o r OILI.ITI:|+’._O>‘IOIO: ..I- s. Iose.11 _ ...IOIOqI. III I 1 A wiwr 111k Wen-Hi aIn std-J F%I44Lu+lis O [II II.._. +.~.lI.II.O..-I.. III H7itolboowen... vueeof seas-ew4ep «a o... 4'e*o a .mw..+v¢e.. a vecI<7I4e e u 14....I. 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