III I I I IIIII III I I I I I II II I I .—I_._I I m II CID—‘03 EIFEEI I}? III III EI‘IZ IIIIII IIIISEI‘I "’I’II..II5’":III‘I IIIII'I3 II: E IE IIIII'IIIIw "I IIIE wfi-‘Iimu III IIIIIII I3, “III II”; . III“ III’ II IIIII IIISII IIII‘II" I III: III IIIII 53-333... III-III IIIIII :I IIJI III. “mask; III III: I383§II§8 III III. II. III III.“ ....III‘EE IIIII‘ILI‘S. ’IY‘JII E III‘I’SE III-ii? ‘ LIBRAR') MichiganSram . U . 'ty gntbéfi —-_—-,... dw- ABSTRACT EFFECT OF VARIETY, NITROGEN FERTILIZATION AND STORAGE TEMPERATURE ON THE CHANGES IN MARC, SUCROSE AND DRY MATTER CONTENT OF SUGARBEETS DURING STORAGE by Roger E. Wyse Three sugarbeet varieties grown with 150 and 75 lbs. of nitrogen per acre were stored for 136 days at 35 and #5 F. A supplementary experiment involved seven varieties. The differences between varieties in degree of marc decomposi- tion, dry matter and sucrose losses and the changes in clear juice purity were determined. At the time of harvest there was a significant dif- ference between varieties in percent marc. Nitrogen fer- tilization had no effect on the marc and sucrose contents or on the clear juice purity. Beets grown under high nitro— gen (lBO#/A) and stored cold (35 F.) were generally higher in percent of original marc over the entire storage period. The variety x treatment interaction was highly significant for both storage temperature and nitrogen fertilization. There appeared to be no correlation between the in- creased unaccounted for impurities in the clear juice and increased impurities in the diffusion juice due to marc solubilization. Roger B. Wyse Beets grown on high levels of nitrogen and stored cold retained the highest percent of original extractable sugar per ton. The decrease in dry matter was greater than the decrease in sucrose over the entire 136 day storage period but particularly in the first 93 days indicating that a significant amount of non-sucrose substances were being reSpired. This may account in part for the increase in clear juice purity which is common in the early stages of storage. A substantial difference was found between the seven experimental varieties in the solubilization of marc during storage. There was also a significant difference in resist- ance to rot, mold and Sprouting between the seven varieties. Solubilization of marc during storage is of interest because it increases the pectin content of the diffusion juice and makes filtration of first carbonation juice dif- ficult. EFFECT OF VARIETY, NITROGEN FERTILIZATION AND STORAGE TEMPERATURE ON THE CHANGES IN MARC, SUCROSE AND DRY MATTER CONTENT OF SUGARBEETS DURING STORAGE By Roger E. Wyse A THESIS Submitted to Michigan State University in partial fulfillment of the requirements for the degree of MASTER OF SCIENCE Department of CrOp Science 1967 ACKNOWLEDGMENTS The author wishes to express his gratitude to Dr. S. T. Dexter for his guidance both in the study and in the preparation of this manuscript. The author also wishes to thank Dr. C. M. Harrison for reviewing the manuscript and to Dr. F. W. Snyder and Mr. M. G. Frakes for help in the analyses. The co-operation of Dr. G. Hogaboam and Mr. J. Neiderer in supplying the sugarbeets was greatly appre- ciated. Finally the author wishes to thank the U.S.D.A. for the grant which made this study possible. ii TABLE OF CONTENTS Page ACKNOWLEDGMENTS ii LIST OF TABLES o o o o o o o 0 V LIST OF ILLUSTRATIONS . vi INTRODUCTION . . 1 LITERATURE REVIEW 3 Sources of Impurities 3 Breakdown of Marc . 3 Pectic Substances as Sources of Impurities 3 Changes in the Pectic Substances During Storage. 4 Effect of Pectic Substances on Processing . 5 Inversion of Sucrose as a Source of Impurities. 6 Sucrose Losses . 6 Sucrose Losses Due to Respiration . 6 Sucrose Losses Due to a Decrease in Purity . 8 MATERIALS AND METHODS IO Analysis . 11 Marc Determination 11 RESULTS AND DISCUSSION. 13 Variables in Marc Determination. 13 Extraction Temperatures 13 Length of Extraction Time. 14 Effect of Nitrogen Fertilization and Variety on Percent Marc at Harvest . . . 17 Variety X Nitrogen Experiment 17 Experimental Varieties. 17 Results of Beet Storage 19 Changes in Percent Marc During Storage 20 Effect of Nitrogen Fertilization . . . 20 Effect of Storage Temperature . . . . . . 2 Effect of Variety . 23 Increase in Impurities in the Diffusion Juice Due to Marc Decomposition. 29 iii Page Relation of Increased Impurities in Diffusion JuiCe to UnaccOunted for Impurities in Clear Juice . . 32 Relationship Between the Changes in Dry Matter, Sucrose and Marc During Storage. . . . 3A Changes in Extractable Sugar During Storage. . . 34 Evaluation of the Method of Analysis . . . . . 39 SUMMARY AND CONCLUSIONS . . . . . . . . . . HO BIBLIOGRAPHY . . . . . . . . . . . . . . 43 APPENDIX . . . . . . . . . . . . . . . A6 iv Tarzl " R) LU C}‘\ ”\1 LIST OF TABLES Page Effect of Extraction Temperature on Percent Marc . . . . . . . . . . . 13 Effect of Varying the Length of Bath Treatment and Extraction Time on Percent Marc . . , . . . . . . . . 15 Percent Marc of Three Varieties at Harvest Adjusted to 25% Dry Matter. . . . . . . 16 Percent Marc of Eight Experimental Varieties at HarVeSt Adjusted to 25% Dry Matter . . . 19 Changes in Marc and Dry Matter Content During 14O Days of Storage in Seven Experimental Varieties . . . . . . . . 20 Impurities Added to the Diffusion Juice by Marc Decomposition Over the 136 Day Storage Period. . . . . . . . . . . 31 The Relationship Between the Impurities Added to the Diffusion Juice by Marc Decomposition and the IncreaSJ in Impurities in the Clear Juice After 136 Days of Storage . . . . . 33 Evaluation of Varieties for Storing Ability Using the Concept CI Extractable Sugar Per Tor of Beets . . . . . . . . . . 36 Evaluation of Nitrogen Fertilization and Storage Temperature Using the Corcept of Extractable Sugar Per Ton . . . . . . . 3o \l? LIST OF ILLUSTRATIONS Effect of Treatment Time on Percent Marc. Effect of Nitrogen Fertilization on Marc Breakdown of Three Varieties During Storage . Effect of Nitrogen Fertilization on Marc Breakdown of Three Individual Varieties Effect of Storage Temperature on Marc Decomposition of Three Varieties During Storage Effect of Storage Temperature on Marc Decomposition of Three Individual Varieties Breakdown of Marc in Three Varieties Over a 136 Day Storage Period. Effect of Storage on the Breakdown of Marc in all Storage Treatments . Relationship Between Percent Marc at Harvest and Loss of Marc During Storage in Seven Experimental Varieties Changes in Percent Dry Matter, Percent Sucrose and Percent Marc of Three Varieties During 136 Days of Storage vi Page 16 21 22 24 25 26 28 3o 35 INTRODUCTION In the past twenty years improved technology has sub- stantially increased the yield of sugarbeets. This coupled with larger acreages and faster mechanical harvest has made it necessary for sugar factories in the United States to lengthen the processing campaign. A longer campaign requires the storage for extended periods of four to five months of a large percentage of the beets delivered. At the time of harvest the sugarbeet is at its peak in processability, but under present storage conditions the quality decreases rapidly after two or three months storage. The primary effects of storage are (l) a substantial de- crease in sucrose, (2) an increase in soluble impurities in the thin juice, (3) an increase in color and lime salts in the thin juice, and (A) an increase in soluble pectic substances in the diffusion juice. The loss in sucrose can be reduced by lowering pile temperatures thus reducing reSpirational losses. The in- crease in impurities increases the production of molasses: an undesirable but tolerable effect. The effect of in— creased lime salts can be counterbalanced by adding soda ash. The increase in pectins however reduces factory effi- ciency by substantially reducing filtration and diffuser flow rates. Previous research on the effects of storage on the soluble pectin content and subsequent decrease in marc on various root and fruit crops has been extensive. In general the results have shown a substantial increase in soluble pectins particularly when stored at warm temperatures. The effect of storage on the breakdown of the insoluble cell wall materials and the subsequent increase in impurities in the diffusion juice in the sugarbeet has yet to be ascer- tained. With the removal of the leaves, petioles, and crown at harvest all accumulation of sucrose and minerals within the sugarbeet root is halted. The composition at this time, therefore, serves as a basis for all subsequent metabolic changes in storage. The important changes which occur in the sugarbeet during storage are those which affect the processability, decrease the recoverable sugar, and cause a solubilization of the marc. These changes are primarily due to respiration and other catabolic processes which increase the amount of soluble impurities and particularly pectic substances, and reduce the sugar content of the beet. This study was aimed at determining the breakdown of the insoluble cell wall materials and the changes occurring in sucrose and dry matter as affected by variety, cultural practices and storage condition. REVIEW OF LITERATURE Sources of Impurities Breakdown pf Marc.--The sugarbeet marc is the insol- uble residue which remains after extraction of the beet with water (McGinnis, 1951). The marc is composed primarily of cellulose (~25%), hemicellulose (~25%), and pectic sub- stances (W5O%). The pectic substances are stable only in cold water and swell and become soluble in hot water (Silin, 1964). The marc averages approximately 5% of the fresh weight of the beet (Silin, 1964). In a test of fifteen varieties, however, Owens at 31. (1954) found the marc content to vary from 3—6%. Pectic Substances as Sources 9f Impurities.-—The pectic substances are high molecular weight polymers, deriv— atives of pectic acid, and made up of galacturonic acid residues. In pectic acid, the carboxyls are free and can readily react with calcium to form insoluble calcium pec— tate. In pectin and protopectin the carboxyls are ester- ified with methanol (Bonner and Galston, 1952). In the sugarbeet the free carboxyls may be as high as 50% methy- lated (Goodban and McCready, 1965). From 6% (Kertesz, 1951) to 30% (Goodban and McCready, 1965) of the hydroxyls in the two and three positions may be acetylated. The pectic substances are in two forms: water soluble and water insoluble. The water insoluble form is proto- pectin, the structure of which is unknown, but which is thought to exist in a matrix complex of pectin, cellulose and hemicellulose. Protopectin upon hydrolysis yields the water soluble pectic acid (Kertesz, 1951 and Joslyn, 1962). The breakdown of protopectin by water is highly tem- perature dependent. Protopectin when treated with hot water swells and gradually dissolves in the form of pectin (Silin, 1964 and Owen, 1955). Silin (1931) extracted dried beet pulp and found only a slight increase in solubility up to 80 C., however, the amount of pectin extracted at 90 C. was 30 times greater than that at 80 C. The soluble forms of the pectic substances are pectin, pectic and pectinic acids (Kertesz, 1951). Pectic and pec— tinic acids have insoluble calcium salts, but are highly esterified and yield methanol and acetic acid salts when heated in alkali solution. Changes in the Pectic Substances During Storage.-—The soluble pectins after an initial increase in the early stages of growth, decrease steadily until harvest. The amount of total pectic substances is decreased by a mois- ture deficiency. Fertilizer has no significant effect (Gaponenkov, 1943). Silin reports that microbial activity and Sprouting increase the quantity of soluble pectins during storage. Walker (1960) found no significant change in the pectin con- tent over a 48 day storage period when beets remained in good condition, but when stored for 161 days at 34 F., the pectin content decreased one—third. If freezing occurs during storage the pectin, which in a normal healthy beet is a large macromolecule, will be decomposed by enzyme action to a colloidal size (Claassen, 1943). Effect 9f Pectic Substances 9n Processing.—-The first noticeable effect of pectic substances in the factory is in the diffuser. Beets which have been exposed to conditions which weaken the protopectin stability must be diffused at lower temperatures to prevent the cossettes from losing their resilience. Loss of resilience will prevent the free flow of water through the diffuser. For frozen or spoiled beets the temperature should not be allowed above 70-75 C. Silin (1964) found a 110% increase in pectic substances in diffusion juices by increasing the temperature from 75.5 C. to 85.1 C. During defecation the pectinic acid present in the diffusion juice is de—esterified, splitting off methanol and acetic acid. The acetic acid forms soluble calcium acetate and the polygalacturonic acid residue forms an insoluble calcium pectate (Silin, 1964 and Goodban and McCready, 1965). h 7' . r. V INA] ff; 1 . .....S.—.... (D Q .,_ . n U [A K: I k.‘ A; LfllCIU OI Sucrcse can be respired to to glucose aid fructo V J. {O C.) kl. amount of irvert sugar tut glocose an mert to l b '13 Rd in I E diffuser FJ. if held at 80 c. for '— ‘1 - M, . - A. 1 deflomQOSC \ '- YU (1 in 1964‘. The decompositi ; .9 I 1 l k) ,1. i C‘ L) which give soluble calcium salt 0 ‘~ C C? ". *7 11L: ._; 1.,» f7. .1 “w- . '__._.. 71 .‘,.‘ .. .. . 1)U_\_iv'\.i’»lff ‘ ' “1 1 h N “ "V rw . 1 N ~-‘ . ’a 1 -- ' ‘ Sugar losses in storage can occ .' " "“ f -. ,h, ,- h 7‘ 1 VJ d 3’23 : kl .1 I '1: C c if ‘ilrectijr‘thrcugrizn; 1rcreat; .1n CD \J O L: o the recoverable sugar. Sugar losses Due to Respiration 1ducts U through respiration and I ies.—— _._ mpurit rclyzed by invertase 1 not only increases the d fructose may be fer- or converted to acids up to 98% of the invert 20 minutes in 2% lime are acids most addition of lime. ur in three primary inversion, P.* thu redu‘ CO C and Inversion.——Loss ajrtmse :iue to respiration can be storage temperatures (3 Parr at al (1940) determined the 7. respiration at various temperature 1— fourid that the amour; c of IV tot of the total I.“ -‘ ',‘. ," \r . I" f _ 111$) L- - x if. _‘1 ma .1. H J .L col apparen substantial particu ilin, 1964 and loss of sugar due s over a 47 day storage *4 evolved accourted I s— + \‘l (‘_ (‘i a! k) 10 es. SUCTOSQ \\ ‘4 1+v Pur_ty percertage accounted for by C02 evolution, however, was not constant tut varied with temperature. for W :i .L + VLJ «.1 l-c. Stcu: (l950) stored beets in large drums at 68-70 F. \ 7 J 45 days and found approximately 80% of the sucrose los. due to 1;? 20% was due to inversion. IV b—r-r 5. J H 90 c‘ }_1 C 93 CL; Poor circulation of air through stored beets can cause build—up of high concentrations of CO2 thus causing anaerobic respiration to take place. Anaerobic respiration 7v— .vYA, L- T h *3, cyly reiuces the sugar content but greatly increases amount of impurities produced (Silin, 1964). Dexter stored beets under water (anaerobically) and in air arctically) at 32 F. and found a decrease of 45 lbs. of .practatle sugar per ton due to an increase in impurities 3,955 mg/lOOS after seven weeks of storage under anaerotic A healthy beet root has a natural immunity against :vasion by saprophytic organisms (Silin, 1964). ”“eecirg, wilting ard bruising, however, will allow invasitu rct and mold organisms. Drought and wilting conditions 7or to harvest may also affect the incidence of rotting storage (Saskili, 1050a, 1950c, and Larmer, 1937). At J- ”‘ih storage temperatures rot and mold will greatly increase cf been made (Nelson, 1950 are Ga ;spiration losses (Barr, 1940, nd Stout, 1950). little research has been published where an evaluatic: yew varieties on the basis of their storing ability has kill, 1950b). However, Smith 01 .1 tutetantial difference between varieties in )r’“, 2‘ ‘ .1 c, \. C) (’+ fr) C1 L r resistance to rot when mother beets were stored in poly- ethylene bags at 38-40 F. Sugar Losses Due tg_a_Decrease in Purity.-—The decrease in recoverable sugar during storage is not solely a result of sucrose lost as CO but is also due to an in- 2: crease in impurities in the thin juice (McGinnis, 1951; Silin, 1964; Carruthers, 1962; and Dexter, 1965). Even if the impurities did not increase, a decrease in sucrose in the clear juice would give a lower purity. In the table prepared by Dexter (1965) from the formula proposed by Great Western Research Lab (Sugar Beet Res., 1964) it can be seen that a 1% decrease in clear juice purity will cause ap- proximately a 6 lb. or 2% decrease in extractable sugar per ton of beets, percentage sucrose remaining the same. In an intensive examination of compositional changes in diffusion juice during storage, Walker (1960) found a decrease in purity from 92.2% to 87.2% in 90 days at 50 F. The apparent impurities calculated from TJP increased from 8,500 to 14,700 mg/lOOS. The only significant compositional change found, however, was an increase in invert at the expense of sucrose with no loss in total sugars. The 6042 mg/lOOS increase in invert was approximately equal to the 6200 mg/lOOS increase in impurities. These results seem very different from those of Barr (1940) and Stout (1950) who accounted for 60 to 80% of their extensive sugar losses by CO evolved and only 20% by inversion. 2 Carruthers (1962) derived the relationship of (3.5 X Na) + (2.5 X K) + (10 X NH2) which accounted for a high percent of the total impurities in fresh beets. Dexter et a1. (1966) using Carruther's factors for Na, K in the clear juice of stored beets found that as 2 the apparent impurities increased particularly at warm and aNH temperatures the percent of impurities unaccounted for by Na, K and aNH decreased sharply. If this relationship 2 is applied to the data presented by Walker the same trend is noted. Dexter proposed that these unaccounted for im- purities may be weak acids with soluble calcium salts formed in the decomposition of sugars and/or cell wall constituents. MATERIALS AND METHODS Three varieties of sugarbeets from a variety nitrogen yield experiment conducted by the Monitor Sugar Co., were stored for 136 days at 32 and 45 F. The three varieties (Commercial, 63-194, and 401) had been planted in 28” rows with nitrogen fertilization rates of high (150#/A) and low (75#/A). The average yields for the three varieties were: Commercial, 23 T/A; 63—194, 18.5 T/A; and 401, 19 T/A. All beets were hand dug and topped by lightly scraping off the petioles and then cutting off the main bud down to a diameter of approximately two inches. Immediately after harvest the beets were bulked by treatment and divided into storage samples of ten beets each. All samples were hand picked to contain only beets of uniform size and to remove broken and diseased beets. The samples were then weighed and placed in mesh bags inside of canvas bags and placed in two walk-in refrigerators at 32—35 F. and 45 F. All samples placed in storage were replicated three times. In order to further study the variations between vari- eties in storage based on marc decomposition, a supplemen- tary experiment was conducted. Eight experimental varieties from a variety test conducted by Dr. George Hogaboam were stored for 140 days at 45 F. All samples were machine 10 11 harvested and badly bruised when placed in storage. In an attempt to prevent rotting the samples were dipped in a 10% Clorox solution prior to storage. Each sample was placed in a plastic bag. Approximately ten samples were then placed in a large canvas bag for storage. Analysis All samples were analyzed for sucrose*, clear juice purity Na, K, a amino nitrogen, mare and dry matter immedi- ately after harvest and at three removal dates approximately 40 days apart. Marc Determination.--Brei samples were taken at the time of sugar analysis and immediately frozen for marc analysis later. For analysis samples were thawed at room temperature and 50 gms of brei weighed into a tared 250 ml beaker and dried at 75 C. for 36 hours in a forced air oven. After 36 hours the samples were reweighed tocietermine the dry matter content. At the same time that the samples were being weighed for drying, two piece buchner funnels were prepared with a 1/4” cellulose filter pad. The filter pads were prepared by pouring a thick slurry of powdered cellu- lose into the funnels which were then pulled down under vacuum and washed with approximately 250 m1 of water. The filters were then dried with the samples for 36 hours and weighed. *Apparent sucrose, polarimetric 12 One hundred seventy-five m1 of distilled water at 80 C. was added to the dried brei in each beaker. The beakers were then placed in an 80 C. 1 0.2 C. water bath for thirty minutes. Samples were stirred several times during the thirty minute hot water treatment. After the bath treatment the samples were poured into the tared two piece buckner funnels. The samples were then extracted with 1000 ml of 80 C. i 3 C. distilled water added in 200 ml aliquots. The filter was pulled to dryness after the addition of each aliquot. Total extraction time including bath treatment was 45 minutes. The funnels were then dried for 24 hours at 75 C., reweighed and the percent marc cal— culated. All values reported for percent sucrose, dry matter, and marc were corrected for shrinkage during storage by multiplying by the ratio of the weight of the sample after storage to the weight of the sample before storage. RESULTS AND DISCUSSION Variables in Marc Determination Extraction Temperatures.--Since the solubility of the pectins which make up approximately 50% of the marc is highly temperature dependent, the percent marc as deter- mined by hot water extraction should also be highly temper- ature dependent. To determine the effects of temperature in the marc determination a well mixed brei sample was divided into 50 g lots and treated in the normal marc extrac— tion procedure except for the following variations in ex— traction temperatures (Table l). All treatments were repli- cated three times. TABLE l.--Effect of Extraction Temperature on Percent Marc. Bath Extraction Treatment Temperature Temperature Pe§cent arc (0.) (C.) l 70 70 4.590 2 80 12 4 . 394 3 80 80 3.844 4 80 100 3.225 The results exemplify the sensitivity of the marc to high extraction temperatures. Increasing both the bath and 13 14 extraction temperatures from 70 C. to 80 C. decreased the percent marc by 0.745. Increasing the extraction tempera- ture from 12-80 C. reduced the percent marc 0.550. The increase from 80 to 100 C. reduced the percent marc by 0.619. These results correlate with the rates of proto- pectin hydrolysis found by Silin (1964). At temperatures below 80 C. the hydrolysis of protopectin was slow, but above 80 C. the breakdown was extremely rapid. In sugar- beet processing, diffusion temperatures do not usually exceed 80 C. The results indicate the necessity of accurate control of bath and extraction temperatures in determining the per- cent marc particularly at temperatures near 80 C. A similar experiment was conducted to determine the effect of the volume of extraction water used. Increasing the volume from 1—2 liters reduced the percent marc 0.158 (3.998+ 3.840). The one liter volume was used in all storage marc determinations to enable a more precise control of extraction temperatures and to reduce the time required for extraction, even though it did not give the lowest per- cent marc. Length_gf Extraction Time.--Solubilization of pectin is not only temperature dependent, but also depends on the length of treatment (Silin, 1964). The extraction time should therefore have an effect on the determination of percent marc. 15 A well mixed brei sample was divided into 50 gm lots and treated with 80 C. distilled water for various bath and extraction times. Table 2 indicates the treatments applied. Lots of six samples were treated in the bath for periods of 10, 30, and 60 minutes after which three samples from each lot were extracted for 10 minutes and three for 20 minutes. The same volume of water was used in both cases. TABLE 2.——Effect of Varying the Length of Bath Treatment and Extraction Time on Percent Marc. Extraction Bath Time Time 10 30 6O 10 4.809 4.485 4,359 20 4.527 4.320 4.205 A rapid decrease in the percent marc was found between the 10 and 30 minute bath times after which the rate decreased (Figure 1). The twenty minute wash removed a greater proportion of the marc (0.28 compared to 0.16) at the 10 minute bath time than at the 30. However, the greater amount extracted in 20 minutes was a constant be- tween 30 and 60 minutes. The results indicate that just over 10 minutes is required for the easily solubilized pectins to become com- pletely soluble and to diffuse out of the cell. With the % Mare (25% DM) 16 4.9-. ”‘71’ 0.28 1 l 10 Min. Wash 4.54- \\ 20 Min. Wash --- \ . \\ O. 4.3.. \{L‘ 4‘ ~ -1 g 0.1538 a I“~~$ 4.1» 10 30 60 Figure l.--Effect Bath Time (min.) of treatment time on percent marc. 1? removal of these pectins the protopectin begins to solubilize at a rate pr0portional to the extraction time. Effect of Nitrogen Fertilization and Variety on Percent Marc at Harvest Variety 5 Nitrogen -—Under the conditions of this ex— periment there was no effect of the rate of nitrogen fer— tilization on the marc and sucrose contents nor on the C. J. P. at harvest. Previous experiments to determine the effects of nitrogen have shown significant differences between high and low levels of nitrogen. In general lower rates of nitrogen increase the percent sucrose and clear juice purity. In order to compare marc values directly between varieties, the percent marc at harvest was adjusted to a constant 25% D.M. to eliminate variations due to differences in dry matter content (Table 3). Although a significant difference was found between varieties in percent marc, the rate of nitrogen fertilization had no effect. Experimental Varieties.——The results of the supple- mentary experiment to determine the variation in percent marc between eight varieties at harvest are given in Table 4. At the time of harvest the marc values ranged from a high of 4.604% in 02 clone to 3.853 in variety D. Varieties D and E are identical genetically except for one inbred line and had essentially identical marc values at harvest. 18 TABLE 3.-—Percent Marc of Three Varieties at Harvest Adjusted to 25% Dry Matter. U U (1) $4 M C.) U60 H CU CZ +3 (1)2 (D 2 CU CC 4—3 +3 "U C) (1)0) mp +3+> 0343 H 008 :5: CCU PC CH 0+3 'Y’DCD (1)2 030) .r.) S—qCU "CO 0 130 C: P0) CUM 54>: "—05-! b0 «45-4 CJG) (Di-4 "CCU n—l ZE—i 1:904 LLQ <04 U) Commercial 150# 4.501 24 656 4.563 75# 4.633 24.310 4.765 Average 4.567 24.483 4.664 63—194 150# 4.826 23.651 5.101 75# 4.759 23 872 4.985 Average 4.793 23.762 5.043 401 150# 4.336 21.824 4.967 75# 4.492 23.012 4.880 Average 4.414 22.418 4.923 Average high nitrogen 4.877 Average low nitrogen 4.876 TABLE 4.——Percent Marc of Eight EXperimental Varieties at Harvest Adjusted to 25% Dry Matter. T r4 . -- 4 Lnadjusted Percent Adjusted Variet Percant Percent y t Dry Matter Marc Marc A 4.240 26.101 4.061 P 4.172 26.850 3.885 c 3.846 24.489 3.926 D 3.944 25.551 3.858 E 4.009 25.341 3.955 F 4.432 26.769 “-139 G 3.837 23.126 4.147 H 4.420 23.997 4.604 — SL(l29 x 133) MS x SP(5822-0) — (SP 6121 x FL 31) Ms x (SP 5822-0) — SL(129 x 133) MS x (SP 6322-0) (SL 129 x SP 6121) MS x (SP 6428—0) - (SL 126 x SP 6121) MS x (SP 6428-0) — (SP 5822—0) SP 6322—0 H — 02 Clone "Ijtiitjotnib l C) I ‘W H w ' I.“ r ‘ Q+I~ ...‘ Rt/f’bUi‘t/S Of 1116261.. Storage After 136 days of storage all samples of the three varieties were in excellent physical condition. The beets stored warm wilted Slightly more than the beets stored cold but no visible rot or mold was found. Excessive desiccation was noted when samples were removed for the first analysis. his problem was alleviated 20 by placing the remaining samples inside plastic bags. (For a complete table of results, see Appendix.) Changes in Percent Marc During Storage Effect of Nitrogen Fertilization.——Figure 2 is an average of all varieties and storage temperatures at the E v . ‘fH'i‘ high and low nitrogen levels. The results indicate a steady decrease in marc over the entire storage period at both nitrogen levels. Although the difference was not signifi- cant the beets grown at the high level of nitrogen were m consistently higher in percent of original marc over the entire storage period. Nitrogen fertilization (Figure 3) had a different effect on each variety. The variety x nitrogen interaction was Significant at the 5% level with an LSD of 0.253. In 63-194 the differences between high and low nitrogen were not significant. However, the commercial variety grown on high nitrogen stored significantly better than those on low nitrogen. In variety 401 just the opposite situation existed with the low nitrogen beets losing a smaller per— centage of their original marc. The results indicate very little effect of nitrogen fertilization on the decomposition of insoluble cell wall material in beets during storage. The differences which did occur were consistent for the whole experiment but varied with individual varieties. However, beets grown on high nitrogen were consistently higher in percent of 21 .mmeOpm wcfiuzc mmfipmfism> mwnzp go czooxmmpn ohms so coaprHHHpnmm cowoppflc mo pomwmmll.m mpsmfim mmeOpm 2H mama mma mm mm 7 L 1 cl '4:— cwmonpfiz 304 I .l I I atmospfiz cmeIII.I.II mmaasmm HH< so new: i mm mm .100 am mm mm :m mm mm mm mm mm OOH oasw Isutfitao JO quaoaeg Percent of Original Marc Percent of Original Marc Percent of Original Marc 22 100 . Commercial Variety 95- \ \ \~.—.—~—-0High Nitrogen 90-- ._4 Low Nitrogen 85 nt- 80 : i e 53 93 136 Days in Storage 100. Variety 63-194 9Si 90' “\. High Nitrogen 85-r Low Nitrogen 80 J. .L 1. 53 93 136 Days in Storage Variety 401 100‘ 95- 90. Low Nitrogen \\ High Nitrogen 857- 80 i t f 53 93 136 Days in Storage Figure 3.--Effect of nitrogen fertilization on marc breakdown of three individual varieties. 23 original marc remaining, indicating that a high level of nitrogen fertilization may contribute to marc stability during storage. Effect 9f Storage Temperature.——Figure 4 is an average of all varieties and nitrogen treatments at each storage temperature. The final difference between the two storage temperatures was Significantly different at the 10% level. The temperature x variety interaction was also significant at the 10% level. A breakdown into individual varieties indicates little effect due to storage temperature in the commercial and 63-194 varieties. Variety 401, however, stored better at 35 F. than at 45 F. Although the final difference between the two tempera- tures was small, the beets stored at 32 F. stored with con- sistently less decomposition of marc than the beets stored at 45 F., particularly in the first 93 days of storage. Individual varieties (Figure 5), showed a significant difference in storing ability. Long term storage may re— quire the proper combination of fertilization, variety and temperature. Effect 9f_Variety.--After 136 days of storage an ana- lysis of variance indicated a significant difference in per— cent marc between the three varieties (Figure 6). Variety MOI, however, did not lose a Significant amount of marc until after 93 days of storage. The other two varieties de- creased in a linear fashion over the entire storage period. 24 mewLOpm wcanso mmeoHnm> moss» mo soapflwooEoomo ohms co magpmnmasou mmeODm Mo pommmmul.: mnzmflm mwAOpm CH mama mma mm mm . w _ mm 0 / I // 11 mm I u I Eommummvmsspgmaeg Eoo// / .Nm IIIIII Amomzv manpmumosme Enmz I /, .lllllll. mmHaEmm HH< no cam: .l// 1 mm .mm 1 OOH oaew Ieutstao JO queoaeg Percent of Original Marc Percent of Original Marc Percent of Original Marc 100 95 9O 85 .. 80 100 95 90 85 80 80 Figure 5. 25 Commercial Variety Cold ‘ " "‘ Warm 53 93 136 Days in Storage Variety 63-194 Cold ---Warm 53 93 136 Days in Storage Variety 401 53 93 136 Days in Storage Effect of storage temperature on marc decomposition of three individual varieties. 26 Percent of Original Marc 87 i - ~w ‘1 i 53 93 136 Days in Storage - 401 -——-— —-——-—-Mean of All Samples Commercial — ———-—————-63-194 Figure 6.——Breakdown of marc in three varieties over a 136 day storage period. 27 Figure 7 is a composite of all marc values at each removal date. The decrease was essentially linear over the ertire storage period. All varieties retained approxi- mately 89% of their original marc. Table 5 shows the changes in marc which occurred over the 140 day storage period for the seven experimental vari- eties stored at 45 F. TABLE 5.-—Changes in Marc and Dry Matter Content During 140 Days of Storage in Seven Experimental Varieties. Percent of Original Marc Harvest Last Removal Remaining at Variety Storage End % 76 96 % 76 % Dry Matter Marc Dry Matter Marc Dry Matter Marc A 26 101 4.240 22.982 3.690 88.05 87.02 B 26.850 4.172 21.747 3.323 80.99 79.65 D 25 551 3.944 22.055 3.358 86.32 85.14 t 25 341 4.009 22.738 3.609 89.73 90.03 F 26.769 4.432 23.948 3.979 89.46 89.77 t 23.127 3.837 21.295 3.363 92.08 87.65 H 23.997 4.420 20.662 3.816 86.10 86.33 Average Loss 13.49 When removed from small amounts of rot storage varieties G and H had only but were excessively sprouted. E had no rot and very few sprouts. completely rotted and was discarded. Variety C was almost The other varieties Variety Percent of Original Marc 100 99 ‘ 98 97 96 95 ‘ 94 93 - 92 ‘ 91 9O 89 88 28 _L L I I ' 53 93 136 Days in Storage Figure 7.-—Effect of storage on the breakdown of marc in all storage treatments. 29 were intermediate between these two extremes but all con- tained excessive rot in the crown area. Before obtaining brei for marc determinations all rotting tissue was removed. A wide range was found within the varieties in the percent of marc solubilized. Variety B losing 20.35% of its origi— nal marc was definitely the inferior variety and variety E T the superior losing only 9.98%. The other varieties were grouped at about 13.49% loss which corresponds rather closely to the 11% average loss in the variety-nitrogen experiment. #3 There appeared to be little connection between origi— nal percent marc and the loss of marc during storage. Fig— ure 8 is a plot of the original percent marc for the seven varieties adjusted to 25% dry matter plotted against the decrease in storage. The two varieties B and D which were low in percent marc had the greater loss in original marc. However, there appeared to be little connection between original percent marc and loss during storage. Pr0per selection of variety for long storage may become a factor to consider in future breeding programs. Increase in Impurities in the Diffusion Juice Due to Marc Decomposition The dry matter in the beet, before or after storage, might be considered to be (I) insoluble marc, (2) sucrose, and (3) solubles other than sucrose in the diffusion juice. A logical means of expressing the effect of marc decomposition on sugarbeet processing is on the increased Original percent Marc 30 4.6~L 4.5«L 4.4.L 4.3 usage ocoomm pmsflm pmo>swm Hm>oEom .Acmwompflz 304 one swam nmmeOPm .m m: not .h mm Moe mmmmo> so soapssae>m--.m mamoEmm .mpomm mm noH Mom nwmzm oanmpomsexm so pmoocoo one meawb mszpwsomede mmmaoom was noflpmwflaflpsom comoapflz mo coapwsam>Muu.m mqm<9 39 high quality beets stored with a smaller decrease in ESPT.* Although the calculation of extractable sugar per ton is an excellent method of combining the factors which enter into the quality of freshly harvested sugarbeets into one number, it does not incorporate the factor of processability. Evaluation 9£_the Method of Analysis Although the accuracy of the procedure was demon- strated in duplicate marc determinations on a large, well— mixed brei sample, the replications in the storage experi- ment occasionally contained marc values which were obviously out of line. However, these variations may have been due to sampling errors and not procedure. At the time of the last removal LSD values of 0.35, 0.25, and 0.439 were required for significance at the 10% level between varieties, inter- actions and treatments reSpectively. The procedure used in marc determination was a rather severe one and may have partially obstructed treatment dif- ferences by removing a large amount of substances which were insoluble prior to the extraction thus blanketing differences. A possible solution would be to reduce the bath temper- ature to 70 C. and wash with cold water. This would also remove the error due to small temperature differences at 80 C. (see pg. 13). *Extractable sugar per ton. SUMMARY AND CONCLUSIONS The effects of sugar beet variety, nitrogen fertili- zation and storage temperature on the decomposition of the insoluble cell wall materials and the changes in apparent sucrose and dry matter in a 136 day storage period were studied. Three varieties-—commercia1, 63—194, and 40l--were grown under high (l50#/A) and low (75#/A) nitrogen fertili- zation and stored at 32-35 F. and 45 F. Percentages of marc, dry matter, apparent sucrose and C. J. P. were deter-, mined at approximately 40 day intervals. MPrc and dry matter were determined at harvest and after 140 days of storage with seven additional varieties to further study the difference in storing ability between varieties. The study yielded the following conclusions: 1. In this experiment only slight differences in sucrose and C. J. P. were found between lots of beets fertilized with 75 vs 150 lbs. of nitrogen per acre. 2. At the time of harvest there was a Significant difference between varieties in percent marc. Nitrogen fertilization had no effect. 3. A rise in temperature from 35 F. to 45 F. sig- nificantly increased the rate of marc 4O 41 decomposition. There was also a significant variety X temperature interaction. The rate of nitrogen fertilization did not affect the solubilization of marc in the overall experiment. However, the variety X nitrogen interaction was highly Significant. In variety 63-194 marc decomposition was the same at both levels of nitrogen. In the commercial variety, however, marc decomposition was Significantly lower at the high nitrogen rate than at the low. Variety 401 was just the reverse of the commercial variety. After 136 days of storage there was a Signifi- cant difference between varieties in the per- cent of original marc lost. Variety 401 was definitely superior in storing ability for the first 93 days of storage. A comparison of seven experimental varieties stored at 45 F. indicated a significant dif- ference between varieties in resistance to rot, mold and Sprouting. The variety most re- sistant to mold retained the highest percent marc. Sprouting appeared to increase solubili- zation of marc. There was little correlation, however, between percent marc at harvest and the decrease in marc during storage. 10. 42 The loss in marc was 11.3 and 13.5 percent in the Variety x Nitrogen and Variety Test reSpectively. There was little correlation between the increase in soluble solids in the diffusion juice due to marc decomposition and the increased impurities in the clear juice. However, no analysis of raffinose and in- vert were conducted to correct for errors in the apparent sucrose values. The absolute decrease in dry matter was Sig- nificantly greater than the decrease in apparent sucrose. This was particularly true at warm storage temperatures, indicating the rate of non-sucrose substrate reSpiration to be substantial. Beets stored cold and beets grown with high nitrogen (150#/A) stored with the least loss of extractable sugar. BIBLIOGRAPHY Barr, 0. G. and Mervine, E. M. 1940. A Preliminary Report on the Effect of Temperature and Beet Condition on Respiration and Loss of Sugar from Beets in Storage. A.S.S.B.T. 3:53-63. Bonner, J., and Galston, A. W. 1952. Principles of Plant Physiology. W. H. Freeman and Company, San Francisco, California. Carruthers, A., Oldfield, J. F. T., and Teague, H. J. 1962. Assessment of Beet Quality. Paper Presented to the Fifteenth Annual Technical Conf. of the British Sugar Corp. Ltd. Claassen, H. 1942. Pectin and Its Influence on Juice Puri- fication and the Filterability of Carbonation Juice. Centr. Zuckerind 50 221, 1942. C.A. 37 6922, 1943. Dexter, S. T., and Frakes, M. G. 1965. Evaluation of Sugar Beet Storage Practices by Using the Percentage Purity of the ”Thin Juice." A.S.S.B.T. 13:607-612. ., Frakes, M. G. and Nichol, G. 1966. The Effect of Low, Medium, and High Nitrogen Fertilizer Rates on the Storagecfi‘Sugarbeet Roots at High and Low Tempera- tures. A.S.S.B.T. 14:147-159. Gaponenkov, T. K. 1940. Zapiski Voronezh. Sel'sko — Kohz Inst. 18(2) 65-72. C.A. 37:1469, 1943. Gaskill, J. O. 1950a. Effects of Wilting, Drought, and Temperature Upon Rotting of Sugar Beets During Storage. A.S.S.B.T. 6:653-659. 1950b. Possibilities for Improving Storage- Rot Resistance of Sugar Beets Through Breeding. A.S.S.B.T. 6 66-669. 1950c. Progress Report on the Effects of Nutri- tion, Bruising, and Washing u on Rotting of Stored Sugar Beets. A.S.S.B.T. 6:6 0—686. Goodban, A. E., and Owens, H. S. 1956. Isolation and PrOperties of Sugar Beet Araban. A.S.S.B.T. 9:129-132. 43 44 Goodban, A. E., and McCready, R. M. 1965. Liming of Sugar Beet Cossettes. A.S.S.B.T. 13:566-575. Joslyn, M. A. 1962. The Chemistry of ProtOpectin: A Critical Review of Historical Data and Recent Develop- ments. Advances in Food Research 11:1-107. Kertesz, Z. I. 1951. The Pectic Substances. Inter- scientific Pub. Inc. New York. Larmer, F. G. 1937. Keeping Quality of Sugarbeets as Influenced by Growth and Nutritional Factors. J. Agr. Res. 54:185-189. McComb, E. A., and McCready, R. M. 1952. Colorimetric Determination of Pectic Substances. J. Anal. Chem- istry 24:1630-1632. McGinnis, R. A. 1951. Beet — Sugar Technology. Reinhold Publishing Corporation, New York. Nelson, R. T. 1950. ReSpiration and Spoilage Studies Employing a Modification of Method Developed by Stout and Fort. A.S.S.B.T. 6:660~663. Owens, H. S., McComb, E. A., and Deming, G. W. 1954. Com- position and Percentage of Marc in Some Varieties of Sugarbeets. A.S.S.B.T. 9:267-271. Owens, H. S., Stark, J. B., Goodban, A. E., and Walker, H. G. Jr. 1955. Application of Compositional Knowledge to Sugar Beet Technology. J. Agr. and Food Chem. 3:350-353. Silin, P. M. and Silina, Z. A. 1931. J. Sugar Ind. 5: 606-611. Silin, P. M. 1964. Technology of Beet - Sugar Production and Refining. (Translated from the Russian, Israel Program for Scientific Translations Ltd.) U.S. Dept. of Commerce. Smith, C. H. 1962. Preliminary Studies of Sugar Beet Root Storage in Polyethylene Bags. U.S.D.A. - A.R.S., Bluebook, pg. 367. Stout, M., and Smith, C. H. 1950. Studies on the Respira- tion of Sugar Beets as Affected by Bruising by Mechanical Harvest, Severing Into Top and Bottom Halves, Chemical Treatment, Nutrition and Variety. A.S.S.S.B.T. 6 670-679. 45 Sugarbeet Res., 1964. Rpt. Determination of Recoverable Sugar Using Formula Proposed by Great Western Research Lab. Denver. U.S.D.A.-A.R.S Bluebook, CR-4—64,pg. 155. Walker Jr., H. G., Rorem, E. S. and McCready, R. M. 1960. Compositional Changes in Diffusion Juices from Stored Sugar Beets. 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