SYMIPTOMIS OF MOLYBDENTUM'I DEFICIENCY IN TOBACCO

ROBERT A. STEINBERG PLANT INDUSTRY STATION, BELTSVILLE, MARYLAND

Received July 17, 1952

No report on the symptoms of molybdenum deficiency in tobacco has appeared in the literature. It seemed desirable, therefore, to determine and describe their nature since the information should be of value in mineral nutrition studies with tobacco. Production of molybdenum deficiency symptoms in Connecticut Broadleaf tobacco was accomplished in water cultures purified by either a modified calcium carbonate or 8-hydroxyquinoline (oxine) nmethod. The type of equipment and fixed-charge technique has been described (4). The few changes miiade for micronutrient studies included use of nutrient solution purification, pure micronutrients, and water condensed from greenhouse heating steam in Pyrex glassware. Pyrex plates were also substituted for the masonite pressboard covers of the 2.5-gallon culture jars. The nutrient solution employed in the calcium carbonate purification experim-ients had the following composition:
A. Water

Ca(N03)2 4 H20 Mg(N03)2 -6 H20 K2HP04 Na2S (saturated) CaCO3

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600.0 ml. 300.0 gm. 70.0 gmi. 9.0 gm. 2 drops 9.0 gm.

B. Water

K2HPO4 (NH4)2S04 NH4H2PO4 Na2S (saturated) CaCO3

750.0 ml. 75.0 gm. 17.5 gm. 6.0 gmi. 2 drops 6.0 gm.

These partial solutions were lheated for one hour at 15 pounds pressure, shaken thoroughly, and let stand overnight before filtering. It was necessary to add the calcium carbonate to solution A in two portions, heating each time in order to approach neutrality. In one run, for example, the final pH of solution A was 7.12 and that of solution B was 6.97. Gaseous ammonia was liberated from solution B during treatmnent. The filtered solutions were each diluted to one liter. Five milliliters of each were added for every 990 ml. of water in the culture jars. A very heavy precipitate was formed on naixing. The method followed in purification with oxine was largely that of HEWITT and JONES (3). Solution A had the following composition: Water, 500.0 m-l.; Ca (NO3)2 4 H20, 150.0 gm.; Ng (NO) 2 6 H2O, 35.0 gin. Solution B had the following composition: Water, 500.0 ml.; KH2PO4, 30.0 gm.; (NH4) 2SO2, 10.0 gm. Each was acidified to pH 1.5 with concentrated hydrochloric acid, 60 mg. of iron as ferric chloride were added, then 24 ml. of oxine solution, and finally one normal KOH gradually with stirring to pH 5.0. The oxine solution consisted of one gram of oxine dissolved in 2.4 ml. of glacial acetic acid
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and diluted to 80 ml. The quantity of oxine added was insufficient to remove all iron. The heavy black precipitate was filtered off after standing overnight and each solution diluted to one liter. Chloroform extraction of residual oxine was omitted. Ten milliliters of each solution together with 0.4 ml. of 1 N of K2CO3 were added for every 980 ml. of water in the culture jars. A slight precipitate was formed on mixing. The micronutrient salts were the purest chlorides available and boric acid. The concentrations employed were Fe, 10.0; Zn, 0.20; Cu, 0.05; Mn, 0.65; B, 0.35, and Mo, 0.05 p.p.m. These quantities were sufficient to produce plants weighing about 60 gm. (air-dried) at flowering during fall and weighing about 150 gm. during spring. Plants grown in the spring usually displayed slight symptoms of iron and nitrogen deficiency if harvest was delayed too long after flowering. Initial acidities of the carbonate purified controls after addition of micronutrients approximated pH 5.8; and those of oxine purified controls were about pH 5.4. Final acidities at harvest were about pH 6.5 and 7.0, respectively. The first indication of molybdenum deficiency was a savoying and mottling of lamina in midleaves about 24 days after the seedlings were placed in the solutions. Bending and twisting of the leaf lamina were usually present and were followed by the appearance of small interveinal necrotic areas that gradually enlarged until the entire leaf was withered. Leaf withering was frequently marginal at the start. These symptoms gradually spread to other leaves until almost all became involved. The molybdenum deficiency achieved was insufficient to cause more than a week of delay in blossoming and an approximately 35% loss in total dry weight. Whereas height of controls averaged 70 inches, that of a deficient plant was about 55 inches. These symptoms were quite different from those of any of the other micronutrient deficiencies (Fe, Zn, Cu, Mn, B) produced in plants grown at the same time. These results were obtained repeatedly in triplicate on omission of molybdenum. The symptoms of molybdenum deficiency, like those of zinc and copper first appeared on the middle leaves of the plant, and only later on the upper and lower leaves. As was also the case with copper, the dead leaf areas became almost white. In zinc deficiency the dead areas were light to dark brown with dark irregular rings and granules. Figure 1 shows an advanced stage of moderate molybdenum deficiency in tobacco. Almost all the leaves were involved, the symptoms including crinkling, twisting, spotting and withering. The molybdenum content in the leaves of a control plant was determined by Mr. Glen Edgington of this station to be about 6.4 p.p.m. when supplied with 10 liters of solution containing 0.05 p.p.m. molybdenum. The concentration in leaves of a deficient plant of the savoy stage was slightly less than 0.3 p.p.m. This value is about six tenths of that suggested as a probable upper limit of the deficiency range in crop plants by STOUT et al. (5). Another check on molybdenum deficiency as the actual cause of these symptoms consisted in the response of cauliflower plants (Early Snowball)

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when grown in the same solutions. Temperatures were too high to permit formation of heads in any of the plants. However, those plants from which molybdenum was withheld displayed severe symptoms of whiptail similar to those described by CLAYTON (2) and WALLACE (6). These consisted in formation of narrow, ruffled leaves with irregular margins, followed still later by leaves with vestiges only of lamina and swollen, tightly compacted bud leaves. Whiptail of cauliflower was found by Clayton to be widespread on unlimed soils of Long Island. AGARWALA (1) found it was necessary to

..,. . .~~~~~~~~~~~~~~~~~~~t

FIG. 1. Symptoms of molybdenum deficiency in Connecticut Broadleaf tobacco.

decrease supply of molybdenum to a maximum of 0.00005 to 0.0005 p.p.m., or less, in order to produce extreme symptoms of whiptail. Through the aid of Dr E M. Roller, molybednum deficiency symptoms in tobacco were found to be associated with high nitrate and low coagulable nitrogen (protein) in the leav es. Soluble organic nitrogen (amino acids) increased only slightly compared to other micronuitrient deficiencies. Cauliflower appeared to require more molybdenum than tobacco. Leaves without lamina are evidence of extreme deficiency in cauliflower. The same level of molybdenum supply (i.e., impurities) produced only moderate symptoms of deficiency in tobacco. On the basis of values determined by Agarwala and the responses obtained in these experiments with both plants, it

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can be estimated that prevention of flowering in tobacco would require that molybdenum supply be decreased to 0.02 to 0.2 parts per billion, or 0.2 to 2.0 micrograms per plant. LITERATURE CITED 1. AGARWALA, S. C. The effect of molybdenum and nitrate status on yield and ascorbic acid content of cauliflower plants in sand culture. Ann. Rept. Agr. and Hort. Sta. Univ. Bristol 1950: 83-89. 1950. 2. CLAYTON, E. E. Investigations of cauliflower diseases on Long Island. New York Agr. Exp. Sta. Bull. 506. 1924. 3. HEWITT, E. J. and JONES, E. W. The production of molybdenum deficiency in plants with special reference to tomato and Brassica crops. Jour. Pomol. and Hort. Sci. 23: 254-262. 1947. 4. STEINBERG, R. A. Influence of acidity, calcium, and magnesium on growth of Xanthi tobacco in water-culture. Plant Physiol. 26: 37-44. 1951. 5. STOUT, P. R., MEAGHER, W. R., PEARSON, G. A., and JOHNSON, C. M. Molybdenum nutrition of crop plants. I. Influence of phosphate and sulfate on the absorption of molybdenum from soils and solution cultures. Plant and Soil 3: 51-87. 1951. 6. WALLACE, T. The Diagnosis of Mineral Deficiencies in Plants by Visual Symptoms. Second edition. H. M. Stationery Office, London. 1951.