u Jgals This is to certify that the thesis entitled \ "Carotene Distribution In The Sweet Potato.‘' presented by Russell H. Ampey has been accepted towards fulfillment of the requirements for Ph. D. ‘1 in Botany 5 Plant Pathology 1 / , i . _ ( l. 1 ’ g ,1. 3 ,- 7 1' \ ’V l, ‘l I. I 4‘ ‘fl‘ 7" .- i). . t l. 1' /|‘ ‘ f / k ' / f‘ ‘ ' 34/1: LII . Major professor Date August 17, 1959 0-169 LIBRARY Michigan State University CAROTENB DISTRIBUTION IN THE SWEET POTATO By Russell M. Ampey AN ABSTRACT Submitted to the School for.Advanced.Graduate Study of Michigan State University of Agriculture and Applied Science in partial fulfillment of the requirements for the degree of DOCIOR OF‘PHILOSOPHY Department of Botany and Plant Pathology 1959 ’ I x / / "x" /. ,1 i . ' . ' , a L , . I! . . _ Approvedfl 4}“fijgjf5; / (. "«*~' » /=<=:; U Russell M. Ampey ABSTRACT CAROTENE DISTRIBUTION IN THE SWEET POTATO The data presented indicate that three sweet potato varieties contained different amounts of carotene. The Queen Mary variety had the highest concentration, the Porto Rico Unit I less, and the Porto Rico variety the lowest concentration. Carotene is not uniformly diStributed throughout the root. The stem-end of the root contained more carotene than the center region and the center region had more than the root-end. Transverse carotene con- centration differences were also indicated with the core usually con- taining more carotene than peelings. Whereas longitudinal difference in the carotene distribution appeared to be characteristic, the trans- verse differences may alternate from.year to year. It appeared that carotene synthesis continued for some time in the sweet potato root after harvest. During storage, the carotene tends to become more uniformly distributed throughout the root. The ratio of the carotene contained in the root-end compared to that contained in the stemrend increased during storage. The peelings of sweet potatoes contained considerable carotene. Discarding this region of the root results in a substantial loss of pro-vitamin.A. The data indicate that the Queen Mary and the Porto Rico sweet potatoes were superior to the Unit I variety in the retention of caro- tene during storage. f V _ a; h V . . . l L . _ a i a r , . _ L , C , ._ l . ‘Ya I _ r 4 , u . xv . ,J. . J a r. u . . » . A l . A r fix . r .. r a, a. , . l A . . n r . r L t r I . c a ,1 . , . . a . I , . a . . n .1. n _ ’ A . n x 7 . , a . . , c . r r . u _ r n A . . r \ , a o _ k t I A (a . . . I » r ‘ r . 7 fl I . . V A t .i; . , a . . l r . _ y y a r i 0 L y. , _ _ r I .x V . CAROTENE DISTRIBUTTON IN THE SWEET POTATO BY Russell M. Ampey A THESIS Submitted to the School of Advanced Graduate Stucbr of Michigan State University of.Agriculture and Applied Science in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY Department of Botany and Plant Pathology 1959 ACKNOWLEDGMENTS The writer is indebted to Dr. H. C. Beeskow, who served as major professor, and to Dr. R. S. Bandurski, Dr. E. J. Benne, Dr. J. E. Cantlon, Dr. W. B. Drew, and Dr. L. W. Mericle who served as a Guidance Committee and read the menuscript. I am also grateful to Dr. E. J. Benne for advice in the analytical methods used and in the designing of several pieces of equipment employed. My sincere thanks also go to Dr. W. D. Baten who assisted me in the statistical analysis. TABLE OF CONTENTS Page Introduction and Literature Review . . . . . . . . . . . . . . 1 Experimental Methods and Materials . . . . . . . . . . . . . 3 Sources of materials . . . . . . . . . . . . . . . . . . 3 Storage . . . . . . . . . . . . . . . . . . . . . . . . . 3 Analytical methods and.procedure . . . . . . . . . . . . h Experimental Results . . . . . . . . . . . . . . . . . . . . . 6 1. Distribution of the carotene within the sweet potato root . . . . . . . . . . . . . . . . . . . . . 6 .A. Stemrend, middle, and root-end regions . . . . . 6 B. ’Peelings and core . . . . . . . . . . . . . . . lO 2. Carotene content of the whole root . . . . . . . . . 12 3. variations in carotene content for crops of two years 13 h. Changes in carotene distribution during storage . . . 15 .A. Changes in carotene content per unit fresh weight during storage . . . . . . . . . . . . . 17 B. Effect of storage on the longitudinal distribution Of carOtene c e o o o o o o a o o o o o a o o 0 l9 (1) Stem—end changes . . . . . . . . . . . . . l9 (2) Middle region changes . . . . . . . . . . l9 (3) Root-end changes . . . . . . . . . . . . . 19 (h) Root-end/stem-end ratio . . . . . . . . . 20 (5) General effects of storage on carotene distribution 0 C . O O C C O C O O O O O O 20 C. Effect of storage on the transverse distribution of carotene . . . . . . . . . . . . . . . . . . 23 (l) Peelings ..... . . . . . . . . . . . . 23 (2) Core . . . . . . . . . . . . . . . . (3) General effect of storage on peelings and core 0 . . O O I C C O O O O O O DiscuSSion O O O 0 O I O O O O O O O O O O O O O O O O O O Carotene content of whole root . . . . . . . . . . . . Stem—end, middle, and root-end differences . . . . Comparison of carotene content of peelings and core the sweet poltato O O C C C O . O O O O O O C O O . O (variation in crops of two different years . . . . . Changes in carotene distribution during storage . . Summary and Conclusions . . . . . . . . . . . . . . . Literature Cited . . . . . . . . . . . . . . . . . . . . Figures . . . . . . . . . . . . . . . . . . . . . . . . Appendix . . . . . . . . . . . . . . . . . . . . . . Weather data . . . . . . . . . . . . . . . . . . . Individual root analyses . . . . . . . . . . . . . in Page 23 2h 26 26 26 29 31 32 36 3? Lo 52 53 5h LIST OF FIGURES Figure Page 1. Average carotene content for stem-end, middle, and root-end of Queen Mary, Porto Rico Unit I, and Porto Rico sweet potatoes . . . . . . . . . . . . . . . Al 2. Carotene content ratios for stem-end, middle, and root- end based on average carotene content of whole root 8%]. to 10m 0 O O O O O O O I O I O O O O O O O I O O h2 3. Carotene content ratios for middle to stem-end and I‘OOt-end t0 Stem-8nd o o o o o o c o o e o o o o o o o 143 h. .Average carotene content of peelings and core regions . uh 5. Average carotene content for Queen.Mary, Porto Rico Unit I, and Porto Rico sweet potatoes . . . . . . . . . hS 6. Carotene content comparison for l9h5 and 19h? crops of the Porto Rico Unit I sweet potatoes . . . . . . . . . h6 7. Effect of storage on the carotene content of the QueenMarysweetpotato................. h? 8. Effect of storage on the carotene content of the Porto Rico sweet potato . . . . . . . . . . . . . . . . AB 9. Effect of storage on the carotene content of the Porto Rico Unit I l9h5 crop . . . . . . . . . . . . . . L9 10. Effect of storage on the carotene content of the Porto Rico Unit I 19h? crop . . . . . . . . . . . . . .. 50 11. The change of the root—end to stemrend ratio during storage to January and after January for Queen Many, Porto Rico and average of two crops of Porto Rico Unit I sweet potatoes . . . . . . . . . . . . . . . . . . . . Sl Table I. III. Iv. VII. VIII. LIST OF TABLES Average carotene content and standard deviations for stemrend, middle, and root—end of Queen Mary, Porto Rico Unit I, and Porto Rico sweet potatoes . . . . . . Analysis of carotene data for regions of Queen Mary Sweet pOtato O O O I O O O O O O O O O O O O O O C O O Carotene content and ratios for stem-end, middle, and root-end based on the average for the root . . . . . . Average carotene content and ratios of the carotene content for middle to stem—end and root-end to stem-end Average carotene content of peelings and core and peelings to core ratio Average carotene content of Queen Mary, Porto Rico Unit I, and Porto Rico sweet potatoes Comparison of the l9h5 and 19h? crops of Porto Rico Unit I sweet potatoes 0 O C O C C . . O C . . C O I 0 Analysis of variance of the carotene content of Porto Rico Unit I for 19h5 and 19h? pertaining to stem-end data Analysis of variance of the Porto Rico Unit I l9bS and 19h? carotene pertaining to middle sections . . . . The carotene content of three sweet potato varieties during two storage periods lEstimate of "correction" in absolute carotene content due to losses in moisture and dry weight during storage .Analysis of carotene data for storage of Queen Mary . . .Analysis of carotene data for storage of Porto Rico . . Ahmalysis of carotene data for Unitll9hSooooooooo storage of Porto Rico Inmalysis of carotene data for storage of Porto Rico Unit I 191*? e o o o o o o o o o o o e o 0' o o o o o o o Page 10 ll 12 lb lb 15 l6 18 21 21 22 22 Table XVI. XVII. XVIII. Page Effect of storage on the carotene content of peelings and core of Porto Rico and Porto Rico Unit I 19h? sweet potatoes 0 O O O O 0 O O O O O O O O O O O O O O O O O 23 .Analysis of carotene data for storage of peelings and core of Unit I 19h? . . . . . . . . . . . . . . . . . 2h .Analysis of carotene data to and after January for peeling to core ratios of Porto Rico Unit I 19h? . . . 25 INTRODUCTION AND LITERATURE RENIEW The sweet potato (Ipomoea Batatas, Lam.) is the most important horticultural crop in the South where it ranks first among the vege- table crops in total acreage, value of production, and value of sales. Montgomery (26) reported that 11,000,000 bushels of sweet potatoes were grown in Louisiana in 1950 with a value of approximately 11,000,000 dollars. The southern market shows preference for "yams“* while the northern market shows preference for sweet potatoes. The sweet potato is an important food item because of several factors, chief among which ' are its high content of total carbohydrates, carotene (pro-vitamin A), and of ascorbic acid (vitamin C). Jacobs (13) in listing the nutritive values of sweet potatoes showed them to contain 125 calories, carotene equal to 7,700 I. U. of vitamin.A and 22 mg. of ascorbic acid per 100 grams. The caroteen contained in the sweet potato is not evenly distri- buted throughout the root. Thus Speirs, Cochran, Peterson, Sherwood, and weaver (32) first reported on the distribution of carotene within the "Unit I Porto Rico" sweet potato variety. They found that there was a progressiVe increase in carotene content from the root-end through the center to the stemrend of the root. Ezell and Wilcox (5) also found regional differences in the carotene content of the "Porto Rico" variety of sweet potato. They found that the stem-end may contain *"Yamfl refers here to the sweet potato Ipomoea Batatas1 Lam. not the true yam.Dioscorea Batatas Decne. The southern commercial market refers to I. Batatas as‘fiyam“ if, when cooked, it is moist and sweet rather than dry and not sweet; the latter is-refered to as sweet po~ tato. The northern commercial market, generally, makes no such distinc- tion. . , _ c a ' , . A r t ‘ ’ ’ ‘ ~ ,- 5 ’ ’ ‘ " ’ _ é - A , O a , ' - - . , - r t . . I . ‘ ,. - x. ’ 1; \A , A I ‘ r r ,‘ ) . - . a 4. ¢ ~ ‘J ,. - I a. L - ' a A \ . . ‘ x ‘ x" 7‘ I \ r ‘ A 5/ 77‘ . r r . . a r , : - r . 7 I ’ , . r v . a A I a ‘w a > 7' r w r. x j .l ,- . y — 9 ‘ 7 g . ‘u i-l) r ’ P r I 2 two to three times as much carotene as the root-end of the same speci- men. There is considerable confusion in the literature regarding the effect of storage on the carotene content of sweet potatoes. Thus Mac— Leod, Armstrong, Heap, and Talbert (18), MacLeod and Utley (19), Ander- son (1), and others, reported increases in carotene content in sweet potatoes during storage. Mitchell and Lease (2S), and Speirs, Cochran, Peterson, Sherwood, and.Weaver (32), and others, however, found no such increases. Changes in the distribution of the carotene within sweet potatoes during storage have so far not been reported. This investigation was designed principally to study the changes in the distribution of carotene during storage in the root of three common sweet potato varieties, the Queen Mary, the Porto Rico Unit I, and the Porto Rico. EXPERIMENTAL METHODS AND MATERIALS "Carotene" as used here includes all fat soluble pigments in the sweet potato of which the predominant pigment is beta-carotene accord- ing to Lease (17), Matlack (20), and Villere, Heinzelman, Pominski, and wakeham (35). Thus Ezell and Wilcox (5) found for the Porto Rico sweet potato that beta-carotene was 82 percent of the total carotenoids. Miller, Melampy, Mikell, and Hernandez (2h) found from.93 to 99 percent beta-carotene of the total carotenoids in the Unit I Porto Rico variety and from 9h to 100 percent beta~carotene of the carotenoids in the Queen Mary sweet potato. Sources of materials. The sweet potatoes used for this study were secured from the Louisiana State EXperiment Station, Baton Rouge, Loui— siana and.Henry‘Hells' farm.in East Baton Rouge Parish, Louisiana. In the present study only varieties commonly grown and available on the open market were used. No attempt was made to secure "genetically pure lines I! 0 Storage. .All sweet potatoes used in this study.had already been cured seven days at a temperature of 80 to 85°F and a relative humidity of 80 percent. Kimbrough (16) reported that healing of wounds result- ing from harvest to be the most important feature in curing. The sweet potatoes were stored in Open crates in a room ambient (60 - 70°F) room temperature until analyzed for carotene. Speirs, Cochran, Peterson, Sherwood, and'Weaver (32) and Bzell and Wilcox (7) found that the moisture content of sweet potatoes remains relatively constant during storage. These authors reported that small )4 losses in weight that may occur during storage are usually due to loss in dry matter and not of water. Speirs M. (33) in a study covering six southern states showed that the Unit I Porto Rico contained 69.8 per- cent water at harvest, 69.2 percent after curing and 69.5 percent after four months of storage; the Queen Mary percentages were 73.7, 72.8, and 73.1 respectively. Since the'moisture content of sweet potatoes re- mains relatively constant during storage the amounts of carotene re- ported in this paper are based upon the fresh weight. Many investiga- tors follow this procedure. Analytical methods and procedure. A one gram sample of fresh root material was found to provide a suitable amount of carotene for readings on the photoelectric colorimeter employed. Samples were takenfrom the stem-end approximately one inch from the scar tissue, the. center of the ‘ root, and approximately one inch from the scar tissue at the root-end. Thin transverse sections were used to make up these one gram samples. When peelings were to be analyzed they were obtained from the center section of the root as were the core samples. A Carotene was extracted from the samples by the Moore and Ely (27) method with minor modifications. One gram of fresh sweet potato tissue was macerated in a Waring Blendor with 25-50 milliliters of commercial acetone for four minutes, washed down with more acetone and macerated for one minute longer. These washings were necessary because of con— siderable foaming. The macerated tissue was filtered through a Bflcrmer funnel. Although the residue was colorless upon filtration it was rinsed further to. insure complete removal of carotene. 5 The filtrate was transferred to a separatory funnel and 50 milli- liters of petroleum ether (c. p. grade) were added and thoroughly mixed by gentle Spinning and turning. ‘Distilled‘water was added to transfer the carotene from the acetone to the petroleum ether. The supernatant petroleum ether solution was washed several times with water to insure the removal of the acetone and it was dried by filtering through an- hydrous sodium sulfate. The solution was brought to a volume of 100 mil~ 1i1iters by the washings which came through the sodium sulfate. The amounts of carotene in the petroleum ether solution were de- termined.by the use of photoelectric filter photometry (Cenco—Sheard- Sanford Photelometer) in which a blue Number 55h Corning glass light filter of standard thickness was employed. The amounts of carotene present in the samples were determined.by the use of a calibration graph prepared by using known concentrations of a pure solution of a mixture of 90 percent beta—carotene and 10 percent alpha-carotene in petroleum ether. EXPERIMENTAL.RESULJS 1. .Distribution of the Carotene Within the Sweet Potato Root .A. Stem-end, middle, and root-end regions Samples from the Stem-end, the middle, the root—end, the peelings, and the core areas of sweet potato roots were analyzed for carotene content. The average amounts of carotene contained in the stemeend, the middle, and the root-end and standard deviations are summarized in Table I and graphically presented in Figure 1. TABLE I AVERAGE CAROTENE CONTENT AND STANDARD DEVIATIONS FOR STEMAEND, MIDDLE, ANDlROOTLEND 0F QUEEN MARY, PORTO RICO UNIT I, AND PORTO RICO SWEET POTATOES Micrograms carotene per gram, fresh weight Variety Stem-end SD Middle SD Root-end SD Queen Mary 268 i 39 167 1.19 121 i 10 Porto Rico Unit I '11; 23h i 32 125 t 10 77 i 7 Porto Rico Unit I '117 177 t 7 1110 t s 102 t 11 Porto Rico 175 .t 12 128 t 10 81 t 11 0n examining the data in Table I it is seen that the averages of the carotene content for the various sections differ fbr the varieties studied. The average carotene content for the stem-end was much larger ' than for the other regions. The average carotene content of the stemeend region was consider- ably larger for the Queen Mary and the Unit I l9h§ than for the Porto Rico or the Unit I l9h7 sweet potatoes. That there was considerable 7 variability in the carotene content of the three varieties can be seen by examining the standard deviations. The difference in carotene content between the regions of the sweet potato root was tested statistically. Table II shows an analysis of variance of the carotene content of the stemrend, the middle, and the root~end of the Queen Mary variety of sweet potatoes. TABLE II ANALYSIS OF CAROTENB DATA FOR REGIONS 0F QUEEN MARY SWEET POTATO Source of variations D.F. S.S. M;Sq. F. Total 19 135,911; Between Regions Average 2 71,716 35,858 9.h9** Within Regions or Error 17 6h,228 3,778 *ISignificant at the one percent level. On testing the averages pertaining to the sections by a Student- ized.Range Table it was found that the average of the carotene measure- ments in the stem—end was significantly larger than the averages of the carotene measurements in the root-end and also larger thanthe averages of the middle section. There was not a significant difference between the average pertaining to the middle and the root~end. This was true for the Porto Rico Unit I 19h5 crop also. By a similar analysis it was found that the averages of the caro— tene contents in all three regions of the Porto Rico variety were sig- nificantly different. This was also found to be true for the Porto Rico Unit I 19h? crop. The present study corroborates the work of Speirs, Cochran, Peter- son, and Weaver (32) and Ezell and Wilcox (5) who showed that the “7 , l 1 , , 1 I x ‘ V\; I I r: r .1 ~ A n - 4‘ , __ . r 4 - x.» fl , __ . t x ’ r ' .. .. _ O . I v 4 ,. , 8 carotene in the Unit I Porto Rico and the Porto Rico sweet potatoes was not uniformly distributed throughout the root. In addition, the present study showed that the Queen.Mary also had similar regional differences and that each region was consistently higher in carotene content than those of the Porto Rico Unit I and the Porto Rico varieties. The data of Speirs et_al. (32) show that the Porto Rico Unit I stem-end contained 128 ug/gm, the center section 118 ug/gm, and the root-end section 108 ug/gn of carotene. The data of Ezell and Wilcox (5) show that these values for the Porto Rico variety were stem-end A5, cen- ter 31, and root-end 19 ug/gm. Table III shows the ratio of carotene contained in the several re- gions of the sweet potato root based on a weighted average computed by combining values for the stem-end, the middle, and the root-end regions. These ratios are graphically presented in Figure 2. TABLE III CAROTENE CONTENT AND RATIOS FOR STEM-END, MIDDLE, .AND.RO0T-END BASED ON THE AVERAGE FOR THE ROOT Micrograms carotene per gram, fresh weight variety Whole Root Stem-end Middle . Root-end. Av. Ratio Av. Ratio Av. Rat1o Av. Ratio Queen Mary“ 181 1.00 268 l.l8 167 0.92 121 0.67 Unit I ’AS 1AA 1.00 ‘23h 1.63 125 0.87 69 0.h8 Unit 1 '117 135 1.00 177 1.31 1110 1.011 87 0.611 Porto Rico 128 1.00 175 1.37 128 1.00. 81 0.63 If the data Of Speirs et a1. (32) were similarly treated the ratios for the Unit I Porto Rico sweet potato were: stem-end section 1.10, the center section 1.02, and the root-end section 0.88. The data of Ezell 9 and Wilcox (5) showed the following for the Porto Rico variety: stem- end 1.36, the center 0.95, and the root-end 0.69. Thus Speirs gt_§1. (32) found that the middle region of the Unit I sweet potato carotene content was 90 percent of that in the stem-end and that the root-end content was 79 Percent of the stem-end content. These percentages for the present study (the average values for the two crops) were lower, being approximately 71 percent for the middle region and 38 percent for the root-end. The percentages for the Porto Rico variety found by Bzell and Wilcox (5) and those for the present study for these regions are quite similar. The percentages as determined by Ezell and Wilcox (5) were middle region 70 percent Of the stem-end con- centration and the root-end £3 percent. The data of the present study Show the middle region concentration to be 70 percent of the stemeend concentration and the root-end to be h6 percent. These percentages for the Queen.Mary variety were 62 and h5 respectively. Table IV shows the averages of the carotene content of the whole sweet potato and their standard deviations, together with the averages of the ratios of the carotene content of the middle region to the caro- tene content of the stem-end and their standard deviations. The table also shows the averages of the ratios of the determinations of the carotene content in the root-end to the stem-end and their standard deviations. These ratios are graphically presented in Figure 3. 0n.examining Table IV it is evldent that there is considerable variation in the distribution of the carotene in the three different sweet potatoes used. For this table a weighted average was not used as in Table III. The ratio of the middle region to the stem-end was larger than the ratio root-end to stem-end for all varieties. The 10 ratios ranged from about 90 percent larger for the Unit I l9h5 for the middle region to the stem-end than the root-end to the stem—end to about A5 percent larger for the same ratios for the Queen Mary variety. TABLE IV AVERAGE CAROTENE CONTENT AND RATIOS OF THE CAROTENE CONTENT FOR MIDDLE TO STEM-END AND ROOT-END TO STEM-END “En51;‘ratata"‘ _ — — ‘M—S:E:Ratio SD R.E. S.e;Ratio_SD_fi_No:__ variety Av.(ug/gm) SD Items Queen Mary 197 t 211 0.77 t 0.06 0.53 '1 0.11 6 Unit I ’115 1113 i 15 ’ 0.65 i 0.07 0.311 t 0.011 19 Unit I 1117 1311 1‘ h 0.81 t 0.02 0.52 i 0.03 76 Porto Rico 125 1-11 0.76 1 0.07 0.118 t 0.06 12 B. Peelings and Core The sweet potato is usually peeled when used for human consumption. The amounts of carotene lost by this practice may be of interest. The literature revealed no information on the carotene content of sweet potato peelings. There are, however, reports for the outer and inner regions of the carrot root. Thus in 19h0 Werner (37) reported higher carotene concentrations in the phloem of carrots than in the xylem. Harper and Zacheile (12) also reported that the phloem contained more carotene than the inner region of the carrot. Since the carrot shows these differences in the carotene content near the surface and within the root it was thought that possibly there were similar differences in the sweet potato root. A transverse slice about one-half inch thick was cut from the center region of the sweet potato root. This disk was peeled approximately as a house wife would 11 peel the root for human consumption. Peelings were analyzed for caro— tene as were also the part remaining - here called the core. Table V shows the carotene content of peelings and core regions of sweet potatoes and their standard deviations. The table also shows the ratio of peelings to core carotene concentration and the standard deviations. Figure A shows graphically the carotene content of peelings and core. TABLE V AVERAGE CAROTENE CONTENT OF FEELINGS AND CORE AND FEELINGS T0 CORE RATIO Micrograms carotene per gram, fresh weight Peelings Core Ratio variety Av. SD Av. SD Peelings/core SD Queen Mary 187 f 15 208 I 21 0.90 f 0.05 unit I '85 ill 1 21 137 :;16 1,05 1 0.35 Unit I 1&7 119 i 7 1A6 1 8 0.82 t 0.18 Porto Rico 113 t i 15 ' 0.8h : 0.13 S 135 It will be noted that the core region of the three varieties con— tained more carotene than the peelings except the l9h5 crop of the Porto Rico Unit I. There was considerable variability in the carotene content as can be seen.by examining the standard deviations. The core region contained about 22 percent more carotene than peelings in the Unit I l9h7 sweet potato and 5 percent less for the same variety in l9h5. The differences between the carotene content of the peelings and the core regions were significantly different for the Unit I 19h? crop, however, they were not significantly different for the other varieties. 12 2. Carotene Content of the Whole Root Relatively dilute solutions are reduired when photoelectric fil- ter photometry is employed to determine carotene concentrations. Either whole sweet potato roots may be run through a food mill and small portions of the pulp be used for samples or the data from the regions of the root might be combined to represent the carotene concentration per unit weight. The latter procedure may result in shorter exposure to the elements and less decomposition prior to extraction of the pig- ment. Thus in the present study the data for the stem-end, the middle, and the root-end samples were combined and used as "the root average" carotene content . Table VI shows the average carotene content for the three var- ieties of sweet potatoes and their standard deviations. The data are graphically presented in Figure 5. TABLE VI AVERAGE CAROTENE CONTENT OF QUEEN MARY, PORTO RICO UNIT I, AND PORTO RICO SWEET POTATOES W Variety Micrograms carotene per gram, fresh weight SD Queen.Mary 197 1 2h Porto Rico Unit 1 19115 1113 i 15 Porto Rico Unit I l9h7 13b 1 h Porto Rico ‘ 135 I ll It will be noted that the Queen.Mary sweet potato contained larger amounts of carotene than the other varieties. This sweet potato var- iety was developed through breeding by the Louisiana State Experiment Station. The Porto Rico variety is an old variety and still favored by 13 many producers.” The Unit I was developed.by selection from the older variety and is the standard commercial variety. These sweet potato varieties have been used in many sweet potato investigations. Ezell and Wilcox (5) found only hh ug/gm carotene for the Porto Rico and.Speirs §£;al, (32) found 38 ug/gm for the same var- iety. The latter investigators also reported 132 ug/gm for the Queen Mary and 80 ug/gm for the Unit I variety. Miller gt_al. (2h) reported 185 ug/gm.beta-carotene for the Queen Mary and 127 ug/gm.for the Unit I, whereas, Fieger et_§l. (8) listed 73 ug/gm.for the Queen Mary and 3h.ug/gm for the Porto Rico Unit I. The amounts of carotene reported in the present study were prac- tically the same as those reported by Miller gt;al. (2h) but were high- er than those reported by speirs EILEE' (32), Ezell and Wilcox (5), and Fieger gt_al. (8). In the present study the amounts of carotene are used only as a basis to evaluate the distribution of carotene within the root and changes of carotene content in the regions of the sweet potato root during storage. 3. variations in Carotene Content for Crops of Two Years Since the Porto Rico Unit I sweet potato is the most popular var- iety in commercial use crops of two different years were analyzed to determine if the distribution of carotene was Similar. Table VII shows the comparison of Porto Rico Unit I sweet potatoes harvested in 19h5 and 19h7 and the comparison is graphically presented in Figure 6. It will be noted that although the total carotene was different for the two crops the concentration of the carotene in both crops was highest in the stem—end, intermediate in the middle and lowest in the 1A root-end. The transverse distribution of the carotene for these two crops, on the contrary, was not similar. The carotene content varied from about 32 percent more in the stem-end region of the 19h5 crop to only 2 percent more in the root- end. The middle region of the 19h5 crop had about 12 percent less caro- tene than the l9h7 crop and the core region about 7 percent less. TABLE VII COMPARISON OF THE 19h5 AND 19N7 CROPS 0F PORTO RICO UNIT I SWEET POTATOES J 1 Micrograms carotene per gram, fresh weight Year Whole Root Stem—end Middle Root-end Peelings Core Av. SB Av. SD Av. SD Av. SD Av. SD Av. SD 1915 113 t 15 23b 1 32 125 t 16 89 7 IAN 1917 13h i h 177 t 7 110 t 5 87 _ h 119 21 137 I 16 7 186 i 8 1+ 1+ l+l+ Table VIII shows an analysis of variance of the carotene content data for the stem-end of the Porto Rico Unit I l9h5 and l9h7 crops. TABLE VIII ANALYSIS OF VARIANCE OF THE CAROTENE CONTENT OF PORTO RICO UNIT I FOR 1985 AND 19N7 PERTAINING TO STEM-END DATA Source of variation D.F. S.S. M.Sq. F. Total 99 73h,775 Between Years 1 51,897 51,897 7.u** Within Years 98 682,878 6,968 ** Significant at the 1 percent level. The F-value in Table VIII indicated that the average of the caro- tene content of the stem-end for l9h5 was significantly larger than 15 that for 19h7 which shows, as far as these measurements are concerned, that the carotene content can vary from.year to year. This may be re- lated to the difference in total rainfall during the growing seasons of l9h5 and 19h7; the former had 16.52 inches, whereas the latter had only 13.23 inches. This variation of 3.29 inches of rain may have contributed to this difference in carotene distribution. The month of July may have been especially critical since the normal precipitation for July is 6.8h but in l9h5 there were 5.03 inches of rainfall and in l9h7 there were only 0.9A or an inch of rainfall.* .A statistical analysis indicated that the average of the carotene content of the middle section for l9h5 is significantly less than that for 19h7. This analysis is shown in Table IX. TABLE IX ANALYSIS OF VARIANCE OF THE PORTO RICO UNIT I 1915 AND 19b? CAROTENE PERTAINING T0 MIDDLE SECTIONS Source of variation D.F. S.S. M.Sq. F. Total 98 190,059 Between Years 1 22,609 22,609 13** within Years 97 167,h50 1,726 **Significant at 1 percent level. A. Changes in Carotene Distribution During Storage .Although there are many references in the literature dealing with the effect of storage on the carotene content of sweet potatoes there are no reports on changes in the distribution of carotene in the root during storage. 16 Samples of the Queen Mary, the Porto Rico, and the Porto Rico Unit I sweet potatoes were analyzed for carotene content from November of 19h6 through July of l9h8. The storage time for each crop was divided into two periods, an early storage period of approximately four months duration (October to January), and a later storage period.running be- yond January. The Queen Mary variety was stored approximately seven lmonths, the Unit I l9h7 crop for ten.months, the Unit I l9h5 crop for thirteen months, and the Porto Rico for approximately sixteen.months. The results of these analyses are shown in Table X and are graph- ically presented in Figures 7, 8, 9, and 10. TABLE X THE CAROTENE CONTENT OF THREE SWEET POTATO VARIETIES DURING TWO STORAGE PERIODS Micrograms carotene per gram, fresh weight Storage Whole SD Stem- SD Middle SD Root- SD RE/SE SD Root End End Ratio QUEEN MARY To January 186 :38 290 1 80 168 1 NO 100 1 10 .311 1' .16 After Jan. 208 1 35 2115 1 26 167 t 28 1112 t 12 .58 1 .11 PORTO RICO To January 130 1 26 200 1 33 125 1 30 65 1 15 .33 it .02 After Jan. 137 1 11. 170 1 17 129 1 12 811 1' 12 .119 1 .07 PORTO RICO UNIT I 19115 To January 161 t 55 27A 1103 160 1 M6 83 i 12 .30 1 .03 After Jan. 1110 I 16 233 1 38 119 1 13 66 1 17 .28 1 .011 PORTO RICO UNIT I 19117 To January lh8 l 7 200 i 11 15h I 7 90 i 5 .h5 l .05 After Jan. 121 I A 1511 1 5 1211 1 17 811 I S .55 1 .011 17 A. Changes in Carotene Content per Unit Fresh Weight During Storage The data for the combined carotene content for the stem-end, middle, and root-end (whole root) show that the Queen Mary and the Porto Rico sweet potatoes had more carotene per unit of fresh weight after prolonged storage. Although the data Showed certain trends these averages were not statistically significant. The Unit I sweet potatoes had less caro- tene per unit of fresh weight after prolonged storage. The differ- ences were statistically significant for the l9h7 crop but were not significant for the l9h5 crop. The lack of statistical signigicance in many cases simply reflects the great variation in the samples taken. .A disturbing factor in analyses of this sort is the fact that there are changes in the moisture and in the dry weight content of sweet po- tatoes during storage. Scott and Matthews (31) studied the loss of weight of sweet potatoes during curing and storage. They found for the Porto Rico sweet potato a weight loss of 1.7 percent during curing, 5.8 percent weight loss for two months of storage, 7.7 percent after four months, and 10.h percent after six months of storage. Their mean values for sixteen sweet potato varieties were 2.1 percent weight loss during curing, 6.7 percent after two months of storage, 9.3 percent after four months, and 13.5 percent after six months of storage. On this basis, the Queen Mary may have lost about 16 percent of its original weight during seven.months of storage, Porto Rico about 26 percent during sixteen.months, Unit I 19h5 about 25 percent during thirteen.months, and the Unit I 19h7 about 22 percent during the ten months of storage. Table XI shows an estimate of "correction" in absolute carotene content due to losses in moisture and dry weight during storage for 18 the combined stem-end, middle, and root-end data (whole root) for the three varieties of sweet potatoes. TABLE XI ESTIMATE OF "CORRECTION" IN ABSOLUTE CAROTENE CONTENT DUE TO LOSSES IN NKESTURE AND DRY WEIGHT DURING STORAGE AS PUBLISHED BY SCOTT AND MATTHEWS (31) ug/gm carotene content on fresh weight basis 816215. After Storage Uncorrected for losses Corrected for losses variety in moisture and dry wt. in.moisture and dry wt. TOIJan. After Jan. 'To Jan. After Jan. Queen Mary 113 186 208 169 175 Porto Rico 130 130 137 120 101 Unit I'h5 125 . 161 lhO lh7 105 Unit I'h7 1A2 lh8 121 135 9b It must be recognized that this loss of weight due to moisture loss and/or loss of dry matter may affect the relative carotene concentration and if there were different losses between regions of the sweet potato root this would affect the relative carotene concentration in these reg- ions. These facts undoubtedly have contributed to the lack of agree- ment among investigators. It must be kept in mind that the data pre- sented in the present paper are based upon the carotene content of sweet potatoes bought by the pound in the market regardless of weight Changes due to either moisture or dry weight losses. 19 B. Effect of Storage on the Longitudinal Distribution of Carotene (1). Stem-end Changes. All of the varieties and.both crops of the Unit I had less carotene per unit of fresh weight in the stem—end after longer storage. The differences amounted to about 18 percent in the Queen Mary, 17 percent in the Porto Rico, 18 percent in the Unit I 19h5, and about 30 percent in the 19h? crop of the Porto Rico Unit I. Due to the great variation between individual analyses these average differences were statistically Significant only for the 19h7 Unit I sweet potatoes. The differences for the Queen.Mary and the 19h? Unit I appear to be larger than the differences which may be due to weight losses. (2). Middle region changes. The middle region of all varieties had less carotene per unit of fresh weight after longer storage than after the short storage period except the Unit I 19h5 crop. The Changes in this region were less than those in the stem-end for the Queen Mary and the Porto Rico there being only a 1 percent difference and a 3 per- cent difference reSpectively. Unit I sweet potatoes, however, had larger differences in this region. The 19h5 crop difference was about 3h per- cent per unit of fresh weight after the longer storage period and the l9h7 crop difference was about 2h percent. These average differences also were statistically significant only for the 19h7 Unit I. The dif- ferences for the unit I sweet potatoes appear to be larger than the differences which may be due to weight losses. (3). Root-end Changes. The carotene content changes in the root- end were not consistent for all varieties. The Queen Mary and the Porto 20 Iiico had a larger carotene Content per unit of fresh weight in this region after prolonged storage than after the shorter period. The per- centage difference was about N2 percent and 29 percent respectively. The Unit I, however, had a lower carotene content per unit of fresh ‘weight in this region after prolonged storage. The differences were 26 percent for the 19h5 crop and 7 percent for the l9b7 crop. However, none of these differences were statistically significant. The fact that losses in carotene content in the Unit I sweet potatoes after pro- longed storage were not significant suggests the effect of loss of dry matter during storage. (A). Root—end/Stem-end ratio. Table X also showed the root—end/ stemsend carotene ratios and their standard deviations. This ratio was larger at the end of the second storage period for the Queen.Mary, the Porto Rico, and the l9h7 crop of the Unit I sweet potatoes. There was a very small decrease in this ratio for the l9h5 Unit I variety. These ratios are graphically presented in Figure 11. (5). General effect of storage on carotene distribution. It was found earlier in the present study that, in general, the data for the stemeend, middle, and root-end carotene content were significantly dif- ferent. The effect of storage upon the longitudinal distribution of the carotene was tested statistically. By the use of the analysis of variance it was found that the longitudinal regional distribution of the carotene was significantly different for both storage periods for all of the sweet potato vari- eties except the short period for the Queen Mary. On testing the dif— ferences between regions for the two storage periods it was found that 21 the Queen Mary regions were not significantly different for either stor- age periods. For the Porto Rico and the Unit I 19A5 crop for the short storage period the regions were not significant but for the longer per- iod they were significantly different. The Unit I l9h7 regions were significantly different for both storage periods. Tables XII through XV Show an analysis of variance for the regions of the sweet potato varieties for the two storage periods. TABLE XII ANALYSIS OF CAROTENE DATA FOR STORAGE OF QUEEN MARY Source of Short Period Long Period variations D.F. S. S. M.Sq. F. D.F. S. S. M.Sq. F. Total 8 1011, 377 8 27,850 . Between Reg. 2 27,1178 13,739 1.1 2 17,011; 8,522 117* within Reg. 6 76,899 12,800 6 10,806 1,801 __N0t significant NSignificant at 5 percent level TABLE XIII ANALYSIS OF CAROTENE DATA FOR STORAGE OF PORTO RICO Source of Short Period Long Period Variations D.F. S. S. M.Sq. F. D.F. S. S. M.Sq. F. Total 5 22,503 29 79,915 Between Reg. 2 18,1111 9,071 6.2* 2 36,656 18,328 11.11“ within Reg. 3 11,362 1,1611 . 27 113,259 1,602 **Significant at 1 percent * Significant at 5 percent level level 22- TABLE XIV ANALYSIS OF CAROTENE DATA FOR STORAGE OF PORTO RICO UNIT I 19h; W Source of Short Period Long Period variations D.F. S.S. M.Sq. F. D.F. S.S. M.Sq. F. Total 8 2hh,281 Ah 5h9,6h9 Between Reg. 2 166,508 83,25N 6.8** 2 176,082 88,0h1 10** Within Reg. 6 77,773 12,929 112 373,567 8,8911 **Significant at 1 percent level **Significant at 1 percent ' level TABLE XV ANALYSIS OF‘CAROTENE DATA FOR STORAGE 0F PORTO RICO UNIT‘I 19h7 Source of Short Period Long Period variations D.F. 8.8. M.Sq. F. D.F. S.S. M.Sq. F. Total 122 591,372 10A 259,821 Between Reg. 2 2N9,753 12h,877 h3** 2 80,197 A0,099 23** Within Reg. 120 3h1,619 2,8h7 102 179,62h 1,761 **Significant at 1 percent level **Significant at l perCent level The averages of the carotene content of the whole root data of the l9h7 crop of the Unit I sweet potato data for the short storage was larger than those data for the longer storage period. T—tests indicate that this difference is significant. This was also true for the stem- end data and those for the middle region but was not true for the root- end. The differences for the "whole root", stem-end, and middle were larger than the differenCes which may be due to weight losses but were not larger for the root-end. 23 C. Effect of Storage on the Transverse Distribution of Carotene Transverse Carotene content differences were found as well as longitudinal differences. The Changes during storage in the carotene content of peelings and core of the Porto Rico and the 19h? crop of the Porto Rico Unit I sweet potatoes and their standard deviations are shown in Table XVI and are graphically presented in Figures 8 and 10. Here, too, the data presented are based upon the carotene content of sweet potatoes bought by the pound in the market regardless of weight changes during storage due to losses of moisture or dry weight. TABLE XVI THE CAROTENE CONTENT OF FEELINGS AND CORE OF PORTO RICO AND PORTO RICO UNIT I 19A? SWEET‘POTATOES DURING TWO STORAGE PERIODS Micrograms carotene per gram, fresh weight Stora e P8811“ 5 Core Peel/Core Ratio 9 P.Rico SD U1 7 SD . ico SD U1 7 SD P.Rico SD UIfh7 SD To Jan. 165 1 5 118 1 9 107 1 19 150 1 9 1.59 1.72 .85 1.09 Aft. Jan. 101 j 16 123 t 7 1A2 I 18 132 1 7 0.76 1.10 .95 1.07 (l). Peelings. It will be noted that the carotene content of peelings 0f the Porto Rico sweet potato appeared to decrease about to percent per unit of fresh weight during prolonged storage. This dif- ference was found to be significant. The Porto Rico Unit I sweet po- tatoes, to the contrary, had increased amounts of carotene per unit of fresh weight after prolonged storage. But this difference was not. statistically significant. (2). Core. The changes in carotene concentration in the core region were also opposite for these two varieties. The change in the 2h carotene content per unit of fresh weight in the Porto Rico core region was approximately 32 percent after the longer storage period. But this difference was not significant. The core region of the Porto Rico Unit I difference for the two periods was about 12 percent. This difference was significant. (3). General effect of storage on peelings and core. By the use of the analysis of variance it was found that the average of the caro- tene determinations for the short storage period for the differences between peelings and core areas were significantly different for the Unit I. An.analysis of the data for the same regions for the longer storage period revealed that they were not significant. The ratio peel/ core was significantly different for data to January and after January storage for both sweet potato varieties. Tables XVII and XVIII show an analysis of the data for storage for the Porto Rico Unit I 19h? crop for peelings and core carotene content. TABLE XVII ANALYSIS OF CAROTENE DATA FOR STORAGE OF FEELINGS AND CORE OF UNIT I 191.7 Source of Short Period Long Period variations D.F. S.S. M.Sq. F. D.F. S.S. M.Sq. F. Total hl 7h,687 9 5,06h Between Reg. 1 10,121. 10,121 6“ 1 221 221 .11 Hithin.Reg. ho 6h,563 1,6lh 8 h,8h3 605 **Significant at l percent level 7 Not Significant By the use of T—tests for peelings/core data for the Unit I 19h? early storage data were significant, however, for both early and late 25 storage, data for peelings and core regions were not significantly dif- ferent. TABLE XVIII ANALYSIS OF CAROTENE DATA TO AND AFTER JANUARY FOR FEELING T0 CORE RATIOS OF PORTO RI00 UNIT I 19117 Source of variations D.F. S.S. M.Sq. F. Total 25 12.3292 Between Periods 1 8.5033 8.5033 53“ Within Periods 2A 3.8259 .1591 **Significant at 1 percent level It is evident that there may be transverse changes in the carotene concentration as well as longitudinal changes. DISCUSSION Carotene content of whole_potato. The three varieties of sweet potatoes which were studied contained different amounts of carotene. The total carotene concentration was highest in the Queen Mary, inter- mediate in the Porto Rico Unit I, and lowest in the Porto Rico. The amounts of carotene in these varieties were found to be higher than those reported by some investigators and lower than those reported by others. 'The absolute amounts of carotene are not so important to the present study, however, as is the use of these values to determine the relative changes in carotene content during storage and the distribution of carotene in the potato. Stem-end, middle, and root—end differences. The results of the present study corroborate those of other investigators in that they in- dicate a longitudinal difference in the carotene concnetration in the sweet potato root. In general, it was found that the stem-end had the highest concentration of carotene, the middle zone intermediate amounts, and the root-end had the lowest concentration. These differences may be associated with either different rates of synthesis and accumulation of carotene within the sweet potato in these different parts or with the traDSport of carotene from the stem after synthesis in the leaves. Several theories have been.postulated for the synthesis of caro- tene by plants. Goodwin (10) states that as far back as 1837 Berzelius considered that the carotenoid pigments were breakdown products of chlorophylls. Willstgtter and Mieg (38) suggested that the alcohol. 2? phytol, one of the components of chlorophyll, might represent a precur— sor in carotenoid synthesis. Bonner (L), however, thought that though phytol may be a precursor in carotenoid synthesis it is formed independ- ently of chlorophyll and.may represent a reduction.product of some com- mon precursor of both phytol and carotenoids rather than the precursor itself. Bandurski (3) suggested a light accelerated synthesis of caro- tene which involved compounds accumulated during previous photosynthetic . activity but was independent of photosynthesis. VHe also found carotene c0uld.be synthesized in the dark in the presence of glucose but synthesis occurred only one fifteenth as rapidly as the correSponding synthesis in light. The carotene in the sweet potato root appears as though it might be tranSported from the leaves with smaller amounts coming from a synthesis within the potato. Carotene is of special interest because it may be converted into vitamin A in the animals body. Roberts and Southwick (30), however, using the electron microscope suggested that vitamin.A.may represent a step in the anabolism as well as the catabolism of carotene bodies and that the carotene bodies, likewise, represent steps in the anabolism and catabolism of chromoplasts. Porter and Lincoln (29) suggested ..." a scheme which is probably valid for all yellow and red carotene containing fruit and.vegetables" where carotene is formed from lycopene. Tbmes, Quackenbush, and.Kargl (3h) found that beta-carotene may be derived from lyc0pene in the to- mato as suggested by Porter and Lincoln but their data did not exclude the possibility that a common precursor could give rise to both beta- carotene and lycopene. 28 Wenzinger (36) suggested that there is no sound basis for assuming that the site of accumulation of carotenoids is also the site of their formation. Johnson and Miller (lb) suggested that in corn the formation of carotenoids in the leaf and formation and/Or storage in the endo- Sperm.are independent processes. This may also be the case for the sweet potato root. Miller and Johnson (23) reported from 196 ug/gm to h95 ug/gm (day basis) of carotene in sweet potato leaves. This is about twice the amount of carotene which is found in the root. Since carotene is not water soluble it would not be translocatable, Goodwin (10), however, states that the carotenoids are almost always solubilized by attachment to proteins. Then the presence of carotene in the root of the sweet po- tato might be attributed to its translocation from the leaves since the light accelerated synthesis of carotene in leaves has been shown to be more rapid than the synthesis in the root. The slow rate of carotene synthesis in the dark may not be sufficient to produce the amounts of carotene which are present in the root. The presence, however, of pre- cursors in the root at harvest may account for the continued dark syne thesis of carotene during storage. If carotene or its precursor were translocated from the sweet potato leaves where most of it is synthe- sized to the root where it accumulates then the gradient from the stem- end to the root-end might be ascribed to this tranSport. Evidence for another possible explanation for the gradient of carotene from the stem-end to the root-end is suggested by Ezell, Wilcox, and Crowder (7). Thay noted that sprouting in the sweet potato occurs first and most profusely near the stem-end and suggested that there may be a corresponding longitudinal difference in metabolic activity. Thus 29 a higher metabolic activity in the stem—end of the sweet potato root may be associated in some way with the higher concentration of carotene in this region. Moore (28), however, cautions against the conclusion that the presence of a high concentration of carotenoid in a particular site necessarily implies that its action in this site is correspondingly intense. Comparison of carotene content of_peelings and of core in the sweet potato If translocation activity were responsible for longitudinal dif— ferences in the concentration of carotene in the sweet potato root one would expect to find a higher concentration near the phloem elements. Thus in studies on the carrot Werner (37) found that from 70 percent to 87 percent of the carotene was in the phloem region. Harper and Zscheile (12) reported as much as 200 percent more carotene in the phloem of the carrot than the inner region. The present study, on the contraty, indicated that in the sweet potato there were larger amounts of carotene in the core region than in the peelings. The difference in the distribution of carotene in the carrot and the sweet potato may be due to the unusual anatomical structure of the sweet potato. .Artschwager (2) and McCormick (21) showed that the epi- dermis of the sweet potato is replaced by periderm and that the stele in the young root is typicall for the dicots. Continued growth and en- largement into fleshy roots, however, give rise to a perplexing anoma- lous structure. Thus, in the older fleshy roots there may be no Single layer of phloem but rather scattered phloem elements including sieve- tubes and companion cells throughout the root. Artschwager (2) found 30 that the larger vascular strands of sec0ndaty tissue may be arranged in either of three ways: (a) the groups are distinct and arranged at the periphery of a narrow circle; (b) the groups are scattered much like the vascular bundles in a corn stalk; (c) the bundles bifurcate, join each other laterally and form in their entirety an intricate network which makes it difficult to trace the individual component parts thoughout the vertical extent of the organ. Some varieties possess characteris- tics of all of these groups. The sweet potato varieties used in the present study appear to have vascular strands of the network type. In all cases secondary phloem tissue (made up of phloem.parenchyma, sieve- tubes, companion cells) is present which lies outside the vascular cams bium. The fact that the carotene content of the core is greater than that of the peelings of sweet potatoes does not preclude the possibility that carotene may be translocated to the roots from the leaves. The role, if any, which carotene plays in the metabolism or plants is not clear. Goodwin (10) reviewed the literature pertaining to the function of carotenoids in.phanerogams under five headings, namely, carotenoids in oxidation—reduction systems, carotenoids as oxygen trans- porters, carotenoids in photosynthesis, carotenodis in photokinetic responses, and carotenoids in reproduction. He concludes that rigorous proof of any Specific function is still lacking. Gortner (11) also believed that there is as yet no proof that carotenoids play a major role in any single physiological mechanism. Frank (9) suggests that phytol may be formed from carotene and thus carotenoids may form a pool for the phytol part of the chlorophyll molecule. 31 variation in crops of two differentgyears. The difference of 9 ug/gm.between the carotene content of the Porto Rico Unit I sweet potato crop of l9h5 and of 19h? was probably due to climatic differences in these two years. As indicated earlier the 19b? growing season had 67 clear days whereas the l9h§ growing season had only DO. The precipita- tion during the l9h7 growing season was less. The month of July es— pecially may have been critical for there were but 0.9h of an inch rainfall, whereas, in l9h5 there were 5.03 inches compared with the normal amount of 6.811 inches.* Goodwin (10) states that it is well established that a period of drought reduces somewhat the carotene con- centratiOn of plants. The l9h5 samples were secured from the Louisiana State Experiment Station farm and the 19h? samples from Henry Wells' farm some ten.miles north of the station. The propagation stock for both crops was from the Experiment Station farm. The soil is claSsed as the same type for both locations. In both crops the stem-end had the highest carotene concentration, the middle intermediate amounts, and the root-end the lowest concentra- tion. The l9h5 crOp had higher average carotene content for the com— bined samples. The l9hS crop also had more carotene in the stem-end and the peelings. However, the 19b? crop had more in the middle, and the core regions. I .Although the total amounts of carotene may be quite different the data show that even in crops of two different years the characteristic longitudinal distribution pattern remains the same. Transverse distri- bution, however, was not the same for both years. *See appendix for weather data. 32 Changes in carotene distribution during storage. Investigators do not agree on the effect of storage on the carotene content of sweet po- tatoes. Ezell and Wilcox (6) and.Anderson (1) presented evidence to show that the carotene content increased during storage, however, Mitch- ell and Lease (25) and Speirs gt_§l. (33) could find no such increases. The loss of weight due to moisture loss and/Or loss of dry matter un— doubtedly have contributed to the lack of agreement among investigators. The data in the present investigation show that there was a great variation in the carotene content of the roots during storage. In the Queen.Mary and the Porto Rico varieties the carotene concentration seem- ed to increase for some time after harvest and then decrease as storage was prolonged. On the contrary, the Unit I variety did not show this initial increase. i ' Queen Mary: The carotene content per unit of fresh weight of the whole root of the Queen.Mary sweet potato changed throughout storage. The stem-end carotene content per unit of fresh weight was considerably less, the middle region change was small, whereas the.root-end had more carotene per unit of fresh weight. The RE/SE ratio was larger for the longer storage than it was for the short storage period. Porto Rico: The carotene content of the whole root of the Porto Rico sweet potato also changed during storage. The stem—end concentra- tion.was less, but the middle and the root~end had.more carotene per unit of fresh weight. The RE/SE ratio also increased with storage time. Porto Rico Unit I: The carotene content of the whole root of both crops of the Unit I had less carotene per unit of fresh weight during storage. All regions of this variety had less carotene after the longer 33 storage period. The difference in carotene content per unit of fresh weight was greater than that which may have been due to low of weight during storage for the 19h? crop. The difference for the l9h5 crop was not larger than that which may have been due to loss of weight during storage. The 19h? crop had a similar increase in the RE/SE ratio as did the other varieties however, the 19h5 crop had only a very slight change for this ratio. Whereas only the Porto Rico Unit I, l9h7, show- ed statistically significant data the other groups indicated only cer— tain trends. It must be recognized that the loss of weight due to moisture loss and/Or loss of dry matter as reported by Scott and Matthews (31) may affect the relative carotene concentration. .An estimate of "correction" in absolute carotene content due to losses in.moisture and day weight during storage was shown in Table XI. If there were different losses between regions of the sweet potato root this would affect the relative carotene concentration in these regions. The data presented in the present paper are based upon the carotene content of sweet potatoes bought by the pound in the market regardless of weight changes due to either mOisture or dry weight losses. The change in the ratio of the carotene contained in the root-end compared to that contained in the stem-end may be due to either trans- port from.the stem-end to the root-end, to differential utilization, to differential destruction or to differential synthesis of carotene in these regions of the root, or to differential moisture and dry weight loss during storage. Transverse carotene concentration differences before and after storage were of smaller magnitude than the longitudinal differences. 311 Two different crops of the Porto Rico Unit I sweet potato had opposite carotene concentration differences in transverse section. These changes in carotene, expressed as peeling to core ratios, were relatively small, however, and often were in the same direction. Although the differences for the Unit I 19h? crop were small the increase of the peelings to core ratio was statistically significant. The-anatomy of the sweet potato root offers avenues of communication.between outer and inner regions if one wishes to assume that carotene as such is tranSported. Superficially it appears that the carotene in sweet potato roots is mobile and is transported along a concentration gradient. However, the carotene may in some way be utilized or destrdyed at different rates in the various parts of the sweet potato or synthesis may occur in var— iable degrees in these regions of the sweet potato root. The fact that the various regions of the sweet potato root contain fluctuating amounts of carotene even after prolonged storage tends to diSprove the possible tranSport of carotene from one region to another. Kehr, Ting, and Mil- ler (lb) concluded from graft union experiments that transfer of caro- tenoids as such from stems of sweet potato plants to the roots did not occur. Until simultaneous analyses of carotene and moisture content and dry weight are made the question of synthesis, distruction or trans- port of carotene are premature. The relatively high carotene content of sweet potato peelings in- dicate that a considerable loss of carotene results from the practice of peeling the root. Thus discarding this region of the root results in the substantial loss of pro-vitamin A. The Queen Mary and the Porto Rico sweet potatoes retained higher percentages of carotene during storage than did the Unit I variety. 35 This was probably due to the greater reduction of carotene in the mid- dle region and in the root-end in the latter variety. The Porto Rico variety is apparently superior to the newer variety, Unit I, for re- tention of carotene during storage. SUMMARY AND CONCLUSIONS 1. (Three sweet potato varieties were analyzed for their carotene content. The Queen Mary sweet potato contained more carotene than the other varieties. The Porto Rico Unit I variety ranked second in caro- tene content and the Porto Rico variety last. 2. The carotene is not uniformaly distributed throughout the sweet potato root. The stem-end contained more carotene than the mid- dle region which, in turn, had more than the root-end. 3. The core region contained more carotene than the peelings. A. Two different crops of the Porto Rico Unit I sweet potato had similar longitudinal distribution of carotene in the root. The trans- verse distribution of the carotene was, however, not the same for the crops of two years. 5. The three sweet potato varieties showed, in general, an increase . in carotene content per unit of fresh weight during early storage and a decrease upon longer storage. 6. The three sweet potato varieties showed, in general, a higher root-end to stem-end ratio of carotene during storage. Changes in the peelings to core ratio were also noted. 7. Considerable carotene is lost when the peelings of sweet po- tatoes are discarded. 8. The Queen Mary and the Porto Rico varieties retained higher percentages of carotene during storage than the Porto Rico Unit I variety. 10. ll. l2. 13. 1h. LITERATURE CITED Anderson, W. S. Some effects of curing and storage on the weight and carotene content of certain sweetpotato varieties. Proc. Amer. Soc. Hort. 801. 68: h12-hl6. 1956 Artschwager, E. On the anatomy of the sweet potato root with notes on internal breakdown. l92h- Jour. Agric. Res. 21:157-166. Bandurski, R. 8. Synthesis of carotenoid pigments in detached bean leaves. Bot. Gaz. 111: 95-109. l9h9. ' Bonner, James Plant Biochemistry. 537 pp. 1950. ‘Ezell, B. D. and Wilcox, M. S. Academic Press Inc., New York. The ratio of carotene to caroten- oid.pigments in sweet—potato varieties. Science 193: l93-l9h. 19A6. Ezell, B. D. and Wilcox, M. S. Effect of variety and storage on carotene and total carotenoid pigments in sweet potatoes. Food Res. 13: 203-212. 19h8. Ezell, B. D., Wilcox, M. S. and Crowder, J. N. Pre- and post- harvest changes in carotene, total carotenoids and ascorbic acid content of sweetpotatoes. Plant Physiol. 211-255-369. 1952. Fieger, E..A., Peterson, W. J. and.Sherwood, F. W. Essentiality of the chromatographing step in the determination of carotene in sweet potatoes. Frank, Sylivia Carotenoids. Sou. Coop. Ser. Bull. No. 10: 97-99. 1951. Sci. Amer. 12g: 80-86. 1956. Goodwin, T. W. Carotenoids their comparative biochemistry. Chemical Publ. Co., New York. 356pp. l95h. Gortner, R. A. Outlines of biochemistry. J. Wiley and Sons, Inc., New York. 1078 pp. Harper, R. H. and.Zscheile, F. P. varieties and strains. 1935. Carotenoid content of carrot Food Res. 19: 8h—97. 19h5. Jacobs, M. B. The chemistry and technology of food and food products. Interscience Publ. Inc., New York. Vol. 2. 1299-1309. 1951. Johnson, I. J. and Miller, E. S. variation in carotenoid pigment concentration among inbred and crossed strains of corn. Cereal Chem. 12: 3115-398. 1955- 15. 16. 17. /18. 19. 20. 21. 22. 23. 2h. 25. 26. 27. 28. 38 Kehr, A. E., Ting, Y. C. and Miller, J. C. The site of carotenoid and anthoqyanin synthesis in sweet potatoes. 'Proc. Amer. Soc. Hort. Sci. 65: 396—398. 1955. Kimbrough, W. D. Curing and storing sweet potatoes without arti- ficial heat. Proc. Amer. Soc. Hort. Sci. 33: h56-h59. 1935. Lease, E. J. Sweet potatoes as a source of vitamin A for man and domestic animals. Assoc. Sou. Agric. Workers. h2nd Ann. Proc. '162—163. 19h1. IMacLeod, F. L., Armstrong, M. R., Heap, M. 'E. and Talbert, L. .A. The vitamin A content of five varieties of sweet potatoes. Jour. .Agric. Res. 50: 181-187. 1935. MacLeod, F. L. and Utley, E. Vitamin.A values. Tennessee Agric. EXP. Sta. Ann. Rept. 51: 6h. 1938. .Matlack, M; B. The carotenoid pigments of the sweet potato (ngmoea Batatas,Poir.). Jour. Washington.Acad. Sci. 21; h93—b95. 1937. McCormick, F..A. Notes on the anatomy of the young tuber of Ipomoea Batatas Lam. Bot, Gaz. 61:7388a398. 1916. Miller, J. C. and Covington, H. M. Some of the factors affecting; the carotene content of sweet poatoes. Proc. Amer. Soc. Hort. jMiller, J. C. and Johnson, R. Study of the carotene content of sweet potato leaves. jRes. In.Agric..Ann. Rept..Agric. Exp. Sta. Louisiana Sta. Univ. 68. l9h5. Miller, J. C., Melampy, R. M., Mickell, J. J. and Hernandez, Th P. Effect of storage on the carotene content of fourteen varieties of sweet potatoes. Proc. Amer. Soc. Hort. Sci. 5g: 399-h02. 19h9. Mitchell, J. H. and Lease, E. S. Stability of carotene in dehy- drated sweet potatoes. Sou. Carolina.Agric. Exp. Sta. Bull. 333. 19A1. Montgomety, J. P. Agricultural statistics for Louisiana. 1909- 1953. ,Louisiana Sta..Agric. Exp. Sta. Louisiana Bull. No. A90. 56. l95h. Moore, L. A. and.Ely, R. Extraction of carotene from plant mater- ial a rapid quantitative method. Ind. End. Chem, Anal. Ed. 13: 600-601. l9hl. Moore, T. Vitamin A. Elsevier Publ. Co., New York. 133 pp. 1957. 29. 30. 31. 32. 33. 3b,. 35. 36. 37. 38. 39 Porter, J. W. and Lincoln, R. E. I. Lycopersicon selections con— taining a high content of carotene and colorless polyenes. II. the mechanism of carotene biosynthesis. Arch. Biochem. 27: 390— 1103 . 1950- Roberts, E..A. and Southwick, M. D. Cantribution of studies with the electron microscope to studies of the relationship of chromo- plasts to carotene bodies and carotene bodies to vitamin A. Plant Physiol. 23} 621-632. l9h8. Scott, L. E. and.Matthews, W. A. Carbohydrate changes in sweet potatoes during curing-and storage. Proc. Amer. Soc. Hort. Sci. 19: 1107—1118. 1957 . Speirs, M., Cochran, H. L., Peterson, W. J., Sherwood, F. W. and Weaver, J. G. The effects of fertilizer treatments, curing, storage, and cooking on the carotene and ascorbic acid content of sweet potatoes. Sou. Coop. Ser. Bull. No. 3. 19b5. Speirs, M., Peterson, W. J., Reder, R., Fieger, E..A., Bheart, J. F., Mitchell, J. H., McLean, F. T., Dempsey,.A. A., Miller, J., Wakeley, J. T., Cochran, F. D., Cordner, H. B., Hollinger, M., James, W. H., Lewis, H., Eheart, M. S., YOung, R. W., Andrews, F. S. and.Garrison, O. B. The effect of variety, curing, storage, and time of planting and harvesting on the carotene, ascorbic acid, and.moisture content of sweetpotatoes grown in six southern states. Sou. Coop. Ser. Bull. No. 30. 3-h8. 1953. Temes, M. L., Quackenbush, F. W. and Kargl, R. E. Synthesis of b-carotene in the tomato fruit. Bot. Gaz. 112: 250-253. 1958. Villere, J. F., Heinzelman, D. C., Pominski, J. and Wakeham, H. R. Isolation of carotene from sweet potatoes. Food Indus. 16: 76—78, 130-131. l9hh. Wenzinger, F. Evolution des pigments carotenoides chez une algue verte. Bull. Soc. de Geneve. 39: 129-21h. 1937. Werner, H. O. Dtyzmatter, sugar, and carotene content of morpho- logical portions of carrots through the growing and storage season. iProc. Amer. Soc. Hort. Sci. 38: 267-272. l9h0. Willstatter, R. and Mieg, w. fiber die gelben Begleiter des FIGURES D STEM END § FRESNO wenem' Q N $5 a us CAROTENE PER 8 QUEEN UNIT l UNIT I PORTO MARY I945 947 RIOO FIGURE I. NERABE OAROT ENE CONTENT OF STEI- END, MIDDLE. All) ROOT—END REGIONS OF THREE SWEET POTATO VARIETIES. ‘SMEN MARY I. IPORTO RICO UNITI I945 PORTO RICO UNITI I947 PORI'O RICO I“I““““‘I I....'..'A I“““““ I.....'......" I“““ STEM-END MIDDLE ROOT-END FIGURE 2. canons: concur RATIOS roe 315w - sun, mucus, mo ROOT-END ”also on AVERAGE canons: counts? or me 3061'. RATIO QUEEN MARY INIT I I945 ' UNIT l I947 PORTO RICO MIDDLE TO STEM~END ROOT- END TO STEM-END FIGURE 3. CAROTENE OWTENT RATIOS FOR MIDDLE T0 STEM- END AND ROOT-END TO STEM‘BID FOR THREE SWEET POTATO VARIETIES. 24 CORE PE ELINCS '5 o - '3 j I 1.16 CAROTENE PER SM, FRESH WEBHT a o QUEEN MARY UNITI I945 UNITI I947 PORTO RICO FIGURE 4. . AVERAGE CAROTENE CONTENT OF FEELINGS AND CORE REGION OF THREE VARIETIES OF SWEET POTATOES‘ .OUE EN MARY NRTO RICO 3 8 no CAROTENE PER on. FRESH WEIGHT 8 'llllllllllllll I\\\\\II\|I\|I FIGURE 5. NERAOE CAROTENE CONTENT OF OIEEN MARY, PORTO RICO INITI, AND PORTO RICO SWEET POTATOES. FRESH [Elsa] 3 3 3 3 MB CAROTENE PER 22 ' 200 '00 I | | |ORE WHOLE STEM- MIDDLE ROOT‘ PEEL ROOT END EM) FIGURE 6. CAROTENE CONTENT COMPARISON FOR I948 AND I947 CROPS OF THE PORT O RICO INITI SWEET POTATOES . 300 — cod. no. 240- DTO JANUARY :20- . AFTER JANUARY '31 FRESH WEIGHT 8 8 8 (”o o 5 0 us CAROTENE PER EH, 0 O O O Q 0 N O WHOLE STEM- MIDDLE ROOT- PEEL CORE ROOT END END FIGURE 7. EFFECT OF STORAGE ON THE CAROTENE CONTENT OF THE QUEEN MARY SWEET POTATO. - TO JANUARY U AFTER JANUARY WHOLE STEM— MIDII.E ROOT- PEEL CORE ROOT END EM) FIGURE 8. EFFECT OF STWAGE ON TI" CAROTENE CONTENT OF THE PORTO RICO SWEET POTATO . m CAROTENE PER GM, FRE$H WEIGHT I 70 JANUARY D AFTER JANUARY WHOLE STEM' MIDILE ROOT- PEEL ROOT END END FIGURE 9. EFFECT OF STORAGE OI THE CAROTENE CMTENT OF THE PORTO RICO UNITI I946 CROP OF SWEET POTATOES. FRESH WEIGHT. 119 CAROTENE PER _ I TO JANUARY 20c. D AFTER JANUARY .5 FRESH WEIGHT 3 5 o 0 GM, 0’ a: — I? 0 o 8 o .5 O 116 CAROTENE PER to o WHG.E STEM- MIDDLE ROOT- PEEL CORE ROOT END END FIGURE IO. EFFECT OF STORfiE GI THE CAROTENE CONTENT OF THE PORTO RICO UNITI I947 CROP OF SWE ET P OTATOES. .5 , I .' I .' I .' .4 o .: g , .‘ - ------ - QUEEN MARY , .3 ...............pom'o moo ,’.' ..._. PORTO RIco UNITI TO JANUARY AFTER JANUARY FIGURE II. THE ORANGE or ROOT-END To STEM- END CAROTENE CONTENT RATIO DURING STORAGE. APPENDIX Month Total precipitation Mean daily Max. temperature Mean Min. Highest daily'Temp. Lowest daily Temp. Clear days Cloudy days Partly cloudy days Cloudiness scale (0-10) Greatest daily precipitation Days measurable precipitation(.25") Month Total precipitation Normal Mean daily Max. temperature Mean Min. Highest daily temp. Lowest daily temp. Clear days Cloudy days Partly cloudy days Cloudiness scale (O-lo) Greatest daily precipitation Days measurable 1985 SUMMER WEATHER SUMMARY* June h-27 91.8 80.9 70.0 96 16th 6h 23rd 12 A IA 5.0 13th 2.18 8 July 5.03 90.5 81.h 72.3 96 17th 69 2lst 8 7 16 5.0 25-26th 1.88 18 19N7 SUMMER WEATHER SUMMARY* May 3.96 -5.31 8h.o 73.8 63.5 90 5th 56 10th 17 h 8 5.5 20th 1.A8 precipitation(.25") A June 53 5.23 A.77 90.0 81.2 71.h 96 12th 65 17th 18 h 8 5.6 18th 2.99 9 AUgust September 3.03 h-l9 91.9 90.8 81.6 79.6 71.3 68.3 98 hth 98 2nd 66 13th St 15th 8 l2 6 1 l7 l7 h.9 3.8 16-l7th 11—12th 0.95 2.02 13 10 July August September 0.9b . 2.21 b.85 6.8A 5.51 h.56 93-3 93.9 91.2 81.h 83.2 79.8 69.6 72.5 68.5 98 28th 100 3lst 99 7th 62 27th 70 lst 53 30th 12 22 15 15 7 13 h 2 2 h.0 h.7 3.h 18-l9th 7-8th 19th 0.2 O.h6 3.79 0 A 5 *Courtesy Harding Field United States Weather Station, Baton Rouge, Louisiana. 5A . DATA FOR INDIVIDUAL ROOT ANALYSES Micrograms of carotene per gram of fresh weight Date Stem-end Middle Root-end Peelings Core QUEEN MARY 11/26/A5 130 100 110 1A0 12/20/A5 360 165 80 155 12/28/A5 380 238 110 235 ... 1/ 5/A6 235 195 130 1A5 16o 1/25/A6 205 109 165 185 220 2/ 9/A6 295 208 130 217 260 A/ 7/A6 155 120 201 190 PORTO RICO 12/31/A5 232 155 80 170 125 12/31/A5 167 95 50 160 88 1/17/A6 157 70 A9 A8 50 A/19/A6 2A3 128 38 118 100 5/2A/A6 190 137 119 155 162 5/2A/A6 195 18A 103 93 190 5/28/A6 177 130 58 100 200 6/ 1/A6 167 110 88 93 100 7/15/A6 1A7 101 6A 6A 170 11/2A/A6 190 200 165 195 200 3/ 8AA? 78 100 59 A5 10A 5/29/A7 155 128 100 ... ... PORTO RICO UNIT I l9h5 11/27/A5 2A0 160 90 120 11/27/A5 115 80 . 60 80 11/27/A5 . 80 A5 12 /28 /A5 A68 2A0 100 1A0 . . . 1/ 2/A6 130 , 155 100 110 1/15/A6 278 170 95 165 210 2/ 1/A6 210 125 90 115 131 3/13/A6 A85 82 70 167 185 3/17/A6 265 115 65 1A0 170 3/85/A6 50 75 20 95 18 / 9/A6 363 165 65 132 260 A/12/A6 85 A7 25 60 A7 A/16/A6 100 100 78 A30 90 A/17/A6 115 70 60 73 80 A/23/A6 383 165 123 155 185 A/2A/A6 551 150 108 163 198 Date Stem-end Middle Root-end Pee lings Core PORTO RICO UNIT I 19A5 (cont.) 10/ 1/A6 172 1A9 50 96 110 10/ 5/A6 . 175 130 1A ... ... 10/11/A6 182 150 58 128 150 10/29/A6 137 12A 55 107 90 11/21/A6 180 92 85 170 150 PORTO RICO UNIT I 19A7 9/ 1/A7 150 -120 93 70 .120 9/ 1/A7 13A 150 50 A7 128 9/ 3/A7 275 190 155 1A8 158 9/ 3/A7 21A 182 6A 170 1A2 9/ A/A7 117 127 50 80 180 9/ 5/A7 158 70 A0 6A 65 9/ 8/A7 200 189 60 115 110 9/ 8/A7 2A0 150 99 155 150 9/ 9/A7 118 97 60 137 1A8 9/ 9/A7 120 138 118 ... ... 9/ 9/A7 157 113 68 78 1A0 9/10/A7 75 6o 50 118 A9 9/12/A7 180 1A3 1A8 ... ... 9/13/A7 218 170 120 90 180 9/1A/A7 267 200 128 1A5 177 9/15/A7 192 1A3 97 127 1A2 9/15/A7 190 1A0 101 ... ... 9/16/A7 202‘ 170 67 160 188 9/16/A7 101 120 153 ‘ ... , ... 9/17/A7 337 229 129 165 222 9/17/A7 21A 95 89 ... * ... 9/18/A7 180 173 73 128 178 9/19/A7 331 199 103 ... ... 9/21/A7 215 16A 93 156 156 9/21/A7 201 ... 109 ... ... 9/23/A7 15A 150 68 102 157 9/2A/A7 208 , 16A 80 ... ... 9/25/A7 229 150 68 ... ... 9/26/A7 1A0 1A7 68 5A 16A 9/27/A7 152 15A 68 ... ... 10/ 1/A7 270 221 123 170 177 10/ 3/A7 231 228 92 ... ... 10/ 3/A7 17A 96 67 10/ 7/A7 370 201 97 ... 10/10/A7 235 175 65 166 10/10/A7 36A 228 A7 70 10/11/A7 2A7 167 107 120 10/1A/A7 193 133 . 78 ... 10/1A/A7 175 162 129 10/18/A7 2A3 21A 132 10/22/A7 22 20 39 Date Stem—end Middle Root—end Peelings Core PORTO RICO UNIT I 19A7 (cont.) 11/ 2/A7 310 170 160 1A8 6/25/A8 108 1A6 ... ... ... 6/25/A8 150 116 68 127. 116 6/26/A8 175 139 78 116 1A6 6/28/A8 170 123 100 116 100 6/28/A8 128 120 85 108 118‘ 6/29/A8 120 132 60 ... ... 6 /30/A8 160 89 128 1A6 . 180 6/30/A8 130 110 - 92 ... ... 7/ 2/A8 16A 150 ... 7/ 9/A8 157 139 89 7/ 9/A8 1A 137 75 7/10/A8 160 137 85 7 /12/A8 12A 1A0 9A 7/12/A8 1A7 89 92 7/13/A8 170 132 128 ... 7/13/A8 200 17A 162 ... 7/1A/A8 160 11A 80 ... 7/15/A8 175 167 103 7/15/A8 1A6 139 23 7/15/A8 186 155 77 7/16/A8 129 ... 78 7/16/A8 15A 128 68 7fl7flfi mm 89 A3 7/17/A8 160 128 115 7/17/A8 199 165 112 7 /17 /A8 132 105 5 9 7 /20/A8 153 12A 103 7/21/A8 150 92 65 7/21/A8 235 150 89 7/21/A8 1A5 123 7A 7/22/A8 123 115 58 7/22/A8 ' 121 7A 53 .7/22/A8 16A 132 78 7/23/A8 82 58 A0 7/23/A8 133 1A6 92 7/23/A8 160 10A 68 ‘9 3:” Q . . I (~ ,. l 8331‘. "‘32.“! ,7... .. Ari m‘flif w .. If IIIIIIIIIII 1762