The Adsorption of a Polygalacturonase Isoenzyme of Botryodiplodia theobromae on Plant Tissues and the Implication on the Pathogenicity of the Fungus

The polygalacturonase isoeazyme (PG 3) of Botryodiplodia theobromae extracted from rotted sweet potato was adsorbed by sweet potato, potato, carrot, bean stem and tomato fruit to various degrees. Adsorption was greater with sweet potato and tomato fruit tissues. Carrots, bean stem and potato absorbed the enzyme to more or less the same degree. The enzyme was not adsorbed on tomato stem. A spore/mycelial suspension of B. theobromae infected the test tissues to various degrees. The enzyme completely macerated sweet potato roots, potato tubers and tomato fruits within 5 h while the bean stem and onion tissues were little affected by the enzyme. The tomato stem was neither infected by the fungus nor macerated by the enzyme.     

The nature and distribution of various forms of nitrogen in the potato

The distribution of nitrogen fractions in the organs of King Edward potato plants harvested at intervals during the growing season has been determined. A general picture of the nitrogen distribution is presented and certain problems suggested by the data are discussed.
Each organ of the plant shows a uniformity in its nitrogenous composition during the whole season of active growth. The seed tubers are characterized by their high content of non-protein nitrogen. This fraction yields the greater part of the nitrogen withdrawn from the seed tubers; the amide, amino and 'other nitrogen' fractions all being markedly depleted. The severely depleted tubers retain some 20% of their initial nitrogen content.
The roots are rich in non-protein nitrogen, in which nitrate nitrogen is an important fraction. The roots from ammonium sulphate-treated plots had a higher total nitrogen content than those from untreated plots, due to accumulation of ammonia nitrogen.
The 'tops' have a higher nitrogen content and a higher proportion of protein than the roots. There is an increase in protein content and a decrease in non-protein nitrogen in passing from stems, to petioles, to leaf laminae. The depleted tubers and roots and stems have a uniform low-protein content of the order 0.4-0.6 mg. protein nitrogen per g. fresh weight.
Asparagine and glutamine occur in approximately equal amounts in the seed tubers and roots. Glutamine is more completely withdrawn than asparagine from the tubers, and in the 'tops' it constantly exceeds asparagine in amount.
A study of the concentration gradients, from leaves to petioles, to stems, to stem bases and new tubers, of the fractions of the non-protein nitrogen focuses attention upon the 'other nitrogen' as containing the organic nitrogen most actively involved in translocation.
The concentration and total content of the glycoside solanine in the different organs has been determined at each sampling.     

NITRATE REDUCTASE IN PHOTOSYNTHETIC BACTERIUM, RHODOSPIRILLUM RUBRUM. ADAPTIVE FORMATION OF NITRATE REDUCTASE

1. A method was discovered for adapting the cells of Rhodospirillumrubrum to grow on a nitrate medium, a capacity initially lackingin the organism. The adapted cells were able to grow with nitrateas the sole source of nitrogen. The growth responses of theadapted cells towards various nitrogenous sources were investigatedunder various conditions of incubation (aero- and anaerobiosis,light and dark).
2. The adapted cells were found to have simultaneouslyacquiredthe capacity for reducing nitrite and hydroxylamineas wellas nitrate. The path of nitrogen in the adapted cellswas assumedto be as follows: NO₃^– NO₂^– NH₂OH CellularNitrogen.
3. Nitrate metabolism of the adapted cells was investigatedundervarious conditions. In the light, nitrate was reducedand furtherassimilated, leaving insignificant amounts of nitritein themedium. In this case, consumption of nitrate was markedlyinhibitedby other forms of nitrogen (e.g., nitrite, hydroxylamine,aminoacids and ammonium salts). In the dark, nitrate was reducedas the terminal hydrogen acceptor in the oxidative breakdownof organic substances (e.g., malate) in the medium (i.e., nitraterespiration). More nitrite was accumulated in this case thanin the light. Molecular oxygen inhibited the reduction of, aswell as the growth on, nitrate in any of the above cases.
4. Theeffects on the rate of nitrate reduction (and respiratoryoxygenuptake) caused by various experimental factors (pH, nitrateconcentration, electron donors, and addition of hydroxylamine)were investigated, using the resting cells of the adapted organism.

¹ This paper was submitted to the University of Tokyo to fulfillthe requirement for the author's doctorate. ² Present Address: Botanical Institute, Kyoto University, Sakyo-ku,Kyoto. (Received February 14, 1963; )     

INFLUENCE OF INORGANIC NITROGENOUS COMPOUNDS ON TOBACCO SEED GERMINATION INDUCED BY GIBBERELLIN A3, KINETIN AND AMMONIUM SALTS OF ORGANIC ACIDS

1. Investigations were made on the influence of inorganic nitrogenouscompounds upon the the germination of tobacco seeds (Nicotianatabacum L. var. virginica (AGDH.) COM. "Bright Yellow") inducedby GA₃, kinetin and ammonium salts of organic acids. Potassiumnitrate, ammonium chloride, ammonium sulfate and ammonium nitrategreatly increase the germination of the seeds induced by theabove reagents, while these inorganic salts, given alone, arealmost ineffective in causing germination.
2. Kinetin was shownto induce germination of tobacco seeds inthe dark. The discrepancywith the results of previous investigationsin this respectwas discussed.
3. It was inferred that nitrogenous metabolismis involved in theprocess of dark-germination of tobacco seedsas induced by theabove-stated stimulating factors and promotedby inorganic nitrogenoussubstances.

(Received July 17, 1961; )     

Increased disease resistance and enzyme activity induced by ethylene and ethylene production of black rot infected sweet potato tissue

Exposure of root tissue from a susceptible variety of sweet potato to low concentrations of ethylene induced a resistance to infection by Ceratocystis fimbriata and an increase in the activity of peroxidase and polyphenoloxidase in the tissue. Susceptible tissue that was inoculated with a pathogenic strain of C. fimbriata or a nonpathogenic strain that can induce resistance liberated more ethylene into closed chambers than tissue inoculated with strains that did not induce resistance. It is suggested that ethylene may be a stimulus that diffuses from infected areas into adjoining tissue to initiate metabolic changes which may lead to disease resistance. Polyphenol oxidase but not peroxidase activity was increased in slices of potato tubers and parsnip roots treated with ethylene. The activity of these enzymes in root tissue of carrot, radish or turnip was not altered by ethylene treatment.     

ADAPTIVE FORMATION OF NITRATE REDUCING SYSTEM IN ANABAENA CYLINDRICA

1. Changes in capacities for reducing nitrate, nitrite and hydroxylaminecaused by provision or depletion of various nitrogen sourceswere investigated with a nitrogen fixing blue-green alga, Anabaenacylindrica, and adaptive nature of these reducing system wasdemonstrated.
2. It was found that, under light-aerobic conditions,nitrate-and nitrite- reducing systems were induced by nitrateor nitritebut not by N₂ ammonia and glutamate. On the otherhand, theactivity of enzymes pertaining to hydroxylamine reductionwasstimulated equally by nitrate, nitrite and N₂. The latteractivitywas suppressed markedly in the presence of ammoniaor glutamate.
3. Adaptive formation of nitrite reducing systemis completelyinhibited by chloramphenicol, a potent inhibitorof proteinsynthesis. No formation was also observed under theanaerobiccondition or in the dark.
4. On the basis of thesefindings, a tentative scheme for pathwaysof nitrate reductionand nitrogen fixation in Analaena cylindricawas proposed.

(Received August 22, 1962; )     

Sucrose Phosphate Synthetase from Various Plant Origins

Some properties of sucrose-P synthetases obtained from various plant tissues, including sweet potato roots, potato tubers and leaves of barley, rape and ladino clover were studied. The specific enzyme activity of the sucrose-P synthetase from sweet potato roots was much lower than that of the sucrose synthetase of the other tissues. The enzyme activity decreased gradually as the roots developed. The optimum pH did not differ between enzyme preparations from sweet potato roots and barley leaves. Manganese chloride exhibited a marked stimulative effect on the sucrose-P synthetase from sweet potato roots and potato tubers, whereas it was inhibited the barley leaf enzyme.

Kinetic studies of sucrose-P synthetase showed that the behavior of the enzyme to the substrates did not differ in the enzyme sources examined. The substrate saturation curve of the enzyme with respect to fructose-6-P was sigmodal in shape, giving a straight line with a slope of 1.35~1.5 (n value) in a plot of the data using the empirical Hill equation. On the other hand, enzymes from all the various tissues exhibited a hyperbolic substrate saturation curve for UDP-glucose, obeying the ordinary Michaelis-Menten type reaction. Manganese chloride had no effect on the Km for UDP-glucose, the S_0.5 for fructose-6-P and the n value of the enzyme from potato tuber tissues.     

SOME PROPERTIES OF PEROXIDASE PRODUCED IN SWEET POTATO INFECTED BY THE BLACK ROT FUNGUS

Chemical and physicochemical properties of peroxidases producedback rotted sweet potato roots were investigated in comparisonwith those produced in cut one. Peroxidases in either diseased or cut tissue were composed offour major (D-A, D-B, D-C and D-D in diseased tissue and C-A,C-B, C-C and C-D in cut tissue) and several minor components.These peroxidases were separated from each other by DEAE-cellulosecolumn chromatography and other procedures. Several propertiesof the peroxidases were investigated.

1. Optimum pH's of peroxidase were in the range of 5.5 to 6.0.
2. The activity of each peroxidase was inhibited by acid, alkaliand some inhibitors such as cyanide, fluoride and azide. Azideinhibited more strongly D-A and C-A than D-B and C-B. On theother hand, cyanide and fluoride inhibited more strongly D-Band C-B than D-A and C-A.
3. Substrate specificity as determimedby using pyrogallol, guaiacol,chlorogenic acid, caffeic acidand umbelliferone differed betweenthe main peroxidases. Thedegree of indoleacetic acid oxidaseactivity of these peroxidaseswas also different from each other.
4. Light absorption spectraof the peroxidases showed that theybelonged to a-type peroxidaseexcept C-D. More precise investigationsof the spectra showedthat the spectra of D-A and C-A were differentfrom those ofD-B and C-B.

Peroxidase A (D-A), the main component in diseased tissue, waspurified by methods such as DEAE-cellulose chromatography andstarchgel electrophoresis to a grade higher than previouslyshown. It was homogeneous, according to investigations withultracentrifugation, immunochemical reaction and starch-gelelectrophoresis. Pyridine hemochrome of the peroxidase showedthat the heme in it was protoheme. Amino acid composition ofthe enzyme was determined. Peroxidase A oxidized various phenolicsubstances in the presence of H₂O₂. Indoleacetic acid oxidaseactivity of peroxidase A was inhibited by both chlorogenic acidand guaiacol. ¹Part 45 of Phytopathological Chemistry of Sweet Potato withBlack Rot. ²Present address: Central Research Institute, Japan MonopolyCorporation, Yutakamachi, Tokyo.     

Response of mycorrhizal seedlings of SW Australian sandplain Epacridaceae to added nitrogen and phosphorus

The responses of seedlings of four species of southwestern Australiansandplain Epacridaceae to added phosphate (P), added NH₄N0₃(N) or Complete nutrients were studied in glasshouse pot cultureusing cores of habitat soil as rooting substrate. Positive responsesto N and Complete nutrients were evident for three species interms of shoot height and shoot dry weight in comparison withControl plants supplied only with deionized water, but no speciesresponded significantly in its shoot growth to P. Root dry weightwas generally less in Complete and N treatments compared toP and Control, leading to considerably higher shoot: root ratiosin the former two treatments. There was no effect of treatmenton infection intensity of hair roots. Root xylem sap compositionshowed greatly elevated levels of nitrogen in the Complete andN treatments and of phosphate in the P treatment. Ammonium comprisedthe major nitrogenous solute of xylem in Control and P treatmentswhile nitrate levels exceeded ammonium in Complete and N treatments.Glutamine levels were particularly high in the P treatments.Labelling of the Complete or N treatments with ¹⁵NH₃ or ¹⁵NO₃(supplied as single labelled ammonium nitrate) indicated thatboth forms of N were taken up and incorporated into plant insolubleN. Key words: Epacridaceae, nitrogen, phosphorus, mycorrhiza, south-west Western Australia     

Development of Heterodera schachtii on Large Rooted Crop Plants and the Significance of Root Debris as Substratum for Increasing Field Infestations

Heterodera schachtii developed to maturity and reproduced on the lateral roots of defoliated sugarbeet which were buried to a depth of 2.5 cm in sterilized soil and inoculated with cysts. Nematodes did not develop on detached lateral roots or on roots of young defoliated beets which did not have a large tap root. The storage roots of large rooted plants were sliced, placed in small jars, inoculated with cysts, covered with moist granulated agar or soil and incubated at 24°C 12-62 days. The sugarbeet nematode developed in root slices of sugarbeet, red table beet, icicle and globe radish, turnip and rutabaga. Only a few males developed on slices of potato tubers. Neither males nor females developed on root slices of carrot, salsify or parsnip. H. schachtii also developed on the cut surfaces of growing sugarbeet and radish.     

MECHANISM OF COLD INJURY IN SWEET POTATOES II. BIOCHEMICAL MECHANISM OF COLD INJURY WITH SPECIAL REFERENCE TO MITOCHONDRIAL ACTIVITIES

1. Respiratory activity of sweet potato root tissues, stored at0° (chilled) and 20° (control), was measured. The activityof the control sample remained approximately constant, whilethat of the chilled one declined sharply after about ten daysof the treatment. Just before this time, DNP-stimulated respiratoryactivity began to drop in the chilled sample, but remained constantin the control.
2. Oxidative and phosphorylative activities ofmitochondria fromthe chilled tissues were compared with thosefrom the control.Exactly as in respiration of the intact roottissues, a sharpdecline in mitochondrial oxidative activitywas found in thechilled tissue after about ten days of thetreatment, but notin the control. In spite of the decreasein oxidative activityof mitochondria, the P/O ratio remainedfairly constant throughoutthe storage. Protein nitrogen ofmitochondria in root tissuesalso remained constant.
3. Contentof polyphenolic substances did not show any significantdifferencebetween the chilled and control tissues.

(Received April 20, 1961; )     

The Uptake and Flow of C, N and lons between Roots and Shoots in Ricinus communis L.

Seedlings of Ricinus communis L. cultivated in quartz sand weresupplied with a nutrient solution containing either 1 mol m^–3NO^–₃ or 1 mol m^–3 NH⁺₄ as the nitrogen source. Duringthe period between 41 and 51 d after sowing, the flows of N,C and inorganic ions between root and shoot were modelled andexpressed on a fresh weight basis. Plant growth was clearlyinhibited in the presence of NH⁺₄. In the xylem sap the majornitrogenous solutes were nitrate (74%) or glutamine (78%) innitrate or ammonium-fed plants, respectively. The pattern ofamino acids was not markedly influenced by nitrogen nutrition;glutamine was the dominant compound in both cases. NH⁺₄ wasnot transported in significant amounts in both treatments. Inthe phloem, nitrogen was transported almost exclusively in organicform, glutamine being the dominant nitrogenous solute, but theN-source affected the amino acids transported. Uptake of nitrogenand carbon per unit fresh weight was only slightly decreasedby ammonium. The partitioning of nitrogen was independent ofthe form of N-nutrition, although the flow of nitrogen and carbonin the phloem was enhanced in ammonium-fed plants. Cation uptakerates were halved in the presence of ammonium and lower quantitiesof K⁺, Na⁺ and Ca²⁺ but not of Mg²⁺ were transported to theshoot. As NH⁺₄ was balanced by a 30-fold increase in chloride in thesolution, chloride uptake was increased 6-fold under ammoniumnutrition. We concluded that ammonium was predominantly assimilated inthe root. Nitrate reduction and assimilation occurred in bothshoot and root. The assimilation of ammonium in roots of ammonium-fedplants was associated with a higher respiration rate. Key words: Ricinus communis, nitrogen nutrition (nitrate/ammonium), phloem, xylem, transport, partitioning, nitrogen, carbon, potassium, sodium, magnesium, calcium, chloride     

Expression of a deregulated tobacco nitrate reductase gene in potato increases biomass production and decreases nitrate concentration in all organs

We investigated the physiological consequences for nitrogen metabolism and growth of the deregulated expression of an N-terminal-deleted tobacco nitrate reductase in two lines of potato (Solanum tuberosum L. cv Safrane). The transgenic plants showed a higher biomass accumulation, especially in tubers, but a constant nitrogen content per plant. This implies that the transformed lines had a reduced nitrogen concentration per unit of dry weight. A severe reduction in nitrate concentrations was also observed in all organs, but was more apparent in tubers where nitrate was almost undetectable in the transgenic lines. In leaves and roots, but not tubers, this nitrate decrease was accompanied by a statistically significant increase in the level of malate, which acts as a counter-anion for nitrate reduction. Apart from glutamine in tubers, no major changes in amino acid concentration were seen in leaves, roots or tubers. We conclude that enhancement of nitrate reduction rate leads to higher biomass production, probably by allowing a better allocation of N-resources to photosynthesis and C-metabolism.Abbreviations DAP Days after planting - Gln Glutamine - NR Nitrate reductase - WT Wild type     

Effect of Mastoparan on Phospholipase A2 Activity in Potato Tubers Treated with Fungal Elicitor

Participation of phospholipase (PL) A₂ in signal trans-ductionhas been reported to elicitate a resistance reaction in potatocells by inoculation of an incompatible race of Phytophthorainfestans, the late blight fungus, or by treatment with fungalelicitor hyphal wall components (HWC). Mastoparan, a genericG protein activator, has been shown to activate PLA in a G protein-dependentmanner in animal cells. We analyzed the effects of mastoparanand the inactive analog Mas-17 on PLA₂ activity in potato tubers.In healthy potato tubers, the activation of PLA₂ by mastoparanwas detected in the soluble fraction, but not micro-somal fraction.However, in potato tubers treated with HWC, PLA₂ activity wasstimulated by mastoparan in both soluble and microsomal fractions.Pretreatment of the microsomal fraction with neomycin, a PLCinhibitor, and staurosporine, a protein kinase inhibitor, inhibitedthe mastoparan-induced activation of PLA₂. This suggested thatthe PLA₂ activation in potato tubers by mastoparan was mediatedby the PLC pathway and protein phospho-rylation. We also examinedthe accumulation of potato phytoalexin rishitin. Mastoparanstimulated rishitin accumulation induced by HWC, but did notinduce the accumulation. This indicated that mastoparan mightactivate the signal transduction pathway in the resistance reactionsinduced in potato tubers. (Received March 12, 1998; Accepted August 6, 1998)     

ORGANIC ACID SYNTHESIS IN RESPONSE TO EXCESS CATION ABSORPTION IN VACUOLATE AND NON-VACUOLATE SECTIONS OF CORN AND BARLEY ROOTS

1. ¹⁴CO₂ fixation into organic acids in tips and proximal sectionsof both corn and barley roots was studied as a function of thenature and concentration of the salt in the external solution.
2. In comparison with the level of ¹⁴CO₂ fixation by vacuolateproximal sections in KCI, incorporation was markedly enhancedin K₂SO₄ and diminished in CaCk. By contrast, non-vacuolateroot tips were indifferent to the type of external salt withrespect to UCO₂ incorporation into organic acids.
3. The effectof salt type on organic acid formation from ¹⁴CO₂was most pronouncedat relatively high concentrations.
4. The conclusion was reachedthat organic acid synthesis in responsetoexcess cation uptakeis the result of cation movement into thevacuole, and thattransport into the vacuole is mediated bythe low-affinity componentof the dual mechanisms involved inion absorption.

¹Present address: Government Forst Experiment Station, Meguro,Tokyo.     

Growth and fat formation of Aspergillus oryzae and Aspergillus terreus on enzyme-hydrolyzed sweet potatoes

Two fungi, namely,Aspergillus oryzae andAspergillus terreus, have been grown on enzyme-hydrolyzed sweet potato medium either as such or supplemented with an external source of nitrogen in the form of ammonium carbonate, sodium nitrate, or asparagine. Hydrolysis alone promoted carbohydrate uptake, growth as well as fat formation to a remarkable extent in both fungi.Addition of an external supply of nitrogen to the hydrolyzed media increased further carbohydrate absorption which continued until the potato media were almost completely depleted; on unhydrolysed potato media, appreciable amounts of carbohydrates always remained unutilized. It also accelerated the rate of building up so that the dry mycelium increased by more than 4 times in some cases. Fat formation was consequently remarkably enhanced specially in the presence of ammonium carbonate or asparagine.     

Studies on plant tissue cultures I. Relationship between inocula sizes and growth of calluses in liquid culture

1. 1. It was observed that lag of growth was longer in small inoculathan in large inocula using tobacco callus in liquid culture.
2. 2. These different growth responses between small and largeinocula were dependent on the ratio of inoculum to culture medium.
3. 3. The same result was obtained in a strain of carrot rootcallus.But the growth lag was very short in the carrot callus,whichwas subcultured for the shortest period among the 4 strainsused, even in small inocula. On the other hand, both small andlarge inocula of the strain, which were subcultured for thelongest period among the 4 strains, did not grow at all duringthe culture period; the longer the period of subculturing, thelonger the lag of growth.
4. 4. The longer lag of small inoculain tobacco callus was recoveredby gibberellin A₃ in the presenceof the acidic fraction ofcarrot root extract or vitamins suchas pyridoxine and thiamine.

(Received December 11, 1967; )     

Microbiological Synthesis of Fat: THE EFFECTS OF NITROGEN LEVEL ON THE METABOLISM OF PENICILLIUM LILACINUM THOM IN A SUCROSE MEDIUM

1. A study has been made of the relationships between the synthesesof carbohydrate, protein, and fat by Penicillium lilacinum Thomin presence of different amounts of sodium nitrate us a definedsucrose salts medium.
2. Under the defined experimental conditionsincreases in the concentrationof NO^–₂ in the medium werefollowed by increases in therates at which nitrogen and sugarwere taken up by the fungus,in the quantities assimilated,and in total and protein nitrogenin the felt. These conditionsprevailed so long as unassimilatedsugar was available.
3. Mediaof lower NO^–₃ concentration (for example, 0·32or 0·64 per cent. (w/v) NaNO^–₂;) yielded feltsricher in carbohydrate than were those grown in media of higherNO^–₂; content (0·96 or 1·28 per cent. (w/v)NaNO₃ The carbohydrate content of the felts increased graduallyuntil the sugar in the medium was exhausted; carbohydrate contentthen decreased.
4. Media of lower NO^–₃; concentration weremore conduciveto fat synthesis than those of higher NO^–₃;content.

     

The Absorption of Ions by Excised Root Systems: II. OBSERVATIONS ON ROOTS OF BARLEY GROWN IN SOLUTIONS DEFICIENT IN PHOSPHORUS, NITROGEN, OR POTASSIUM

1. Barley plants were grown in complete culture solution and indeficiencies of phosphorus, nitrogen, or potassium for a periodof about 6 weeks. Excised roots of these plants were treatedwith a complete, aerated culture solution at 25? C. for varyingperiods of time, and the changes in respiration rate, phosphorus,nitrogen, potassium, sugars, and starch contents measured.
2. Therewere changes in fresh weight and dry weight of the excisedrootsduring treatment. The dry weight decreased with time butthewater-content changes were variable. There was a gain orlossof water by the roots according to the treatment.
3. In all casesthe deficient roots increased in content of theelement in whichthey were originally deficient. The roots ofthe plants suppliedwith full nutrient usually decreased incontent of phosphorus,nitrogen, and potassium, but exceptionsoccurred and the reasonsare discussed.
4. In most of the experiments described simultaneousloss of oneion and gain of another occurred.
5. Nitrogen-deficientroots accumulated nitrate when exposed toa complete nutrientsolution, and some of this was assimilatedwith formation ofprotein. Under similar conditions nitrogen-richroots decreasedin nitrogen content and proteolysis took place.
6. There wasa rapid fall in sucrose and reducing sugar contentof the excise'roots. The starch content was initially verysmall and showedlittle change with time.
7. The respiration rate declined withtime in all treatments exceptwhere a nitrogen deficiency existed.Here the respiration rateincreased to a maximum value at about8 hours and then fell.This increase in rate is attributed toprotein synthesis. Noevidence of a ‘salt respiration’was observed evenwhen active uptake of phosphorus or potassiumwas occurring.
8. In most instances the carbon dioxide evolvedin respirationgreatly exceeded the carbon dioxide equivalentof the sugarconsumed in the same period. Exceptions were foundwith thenitrogen-deficient roots where less carbon dioxidewas evolvedthan the equivalent of sugar consumed. It is probablethat apart, at least, of the sugar unaccounted for was usedin proteinsynthesis.
9. Where the carbon dioxide of respirationwas in excess of theequivalent of sugar consumed, protein oramino-acid is the mostprobable substrate. Respiration rateis found to be relatedboth to nitrogen and sugar content.

     

INTERACTION OF GIBBERELLIN A3, AND INORGANIC PHOSPHATE IN TOBACCO SEED GERMINATION

1. Investigation was made on the influence of inorganic phosphateupon the germination of positively photoblastic tobacco seed(Nicotiana tabacum L. var. uirginica (AGDH.) COM. "Bright Yellow")induced by GA₃, GA₃M, kinetin, red light, and ammonium saltsof various organic acids.
2. Inorganic phosphate increases theGAs-induced germination, andinhibits the germination causedby ammonium citrate, while itdoes not influence the germinationbrought about by GA₃M, kinetin,and red light.
3. The optimumpH for the GA₃-induced germination lies in the acidicpH range,indicating that the undissociated form of GA₃ is operative.The stimulatory effect of phosphate is, however, not ascribedmerely to the pH control in the mediurr. Phosphate exerts somespecific influence for which the presence of the free carboxylgroup of GAs is required.
4. The observed contrasting effectsof phosphate on the GA₃-inducedgermination (i.e., acceleration),on the one hand, and on theammonium citrate-induced germination(i.e., inhibition), onthe other, were explained by assumingthat the phosphate effectsultimately consist in acceleratingthe uptake of the carboxylicacid into the seeds.
5. GA₃M alsohas an activity of inducing the germination of tobaccoseedwithout light.

¹Present address: Department of Vegetable Crops, Universityof California, Davis, California, U.S.A. (Received March 12, 1962; )