The Presence of Gibberellins in Excised Tomato Roots

Substances having similar physiological properties to the gibberellins(located by the ‘Meteor’ dwarf pea bio-assay) havebeen detected in extracts from excised tomato root. The chromatographicbehaviour of the most active zone is similar to that of gibberellinA₁. Experiments using the d-1 and d-5 mutants of mazie did not indicatethe presence of substances with differential effects on thesetwo mutants.     

The Effects of l-Naphthylacetic Acid on the Growth of Excised Tomato Roots

I-naphthylacetic acid (NAA) enhances main axis extension andlateral initiation of excised tomato roots cultured in 0.5 percent sucrose medium. NAA has a qualitatively similar effectupon the growth of root tips cultured in glucose medium or transferredafter 2 days' previous growth in standard (1.5 per cent sucrose)medium to a glucose medium containing NAA.NAA does not raisethe growth-rate of roots cultured in glucose medium to thatoccurring in presence of sucrose as carbon source. The enhancedgrowth resulting from addition of NAA to glucose medium is notdependent upon enhanced glucose uptake.The unique role of sucrosein the growth of excised tomato roots is discussed in the lightof the above observations and of studies on the ability of glucoseto maintain the growth of excised roots previously establishedby a period of culture in sucrose medium.     

Effects of 3-Indolylacetic Acid and 3-Indolylacetonitrile on the Growth of Excised Tomato Roots

1. The inhibition by IAA (3-indolylacetic acid) and by IAN (3-indolylacetonitrile)of the growth of excised tomato roots cultured for 7 days at27 C. in a modified White's medium is described. 510^–9g./ml, IAA or 510^–6 g./ml, IAN cause approx, 50 per cent,inhibition of the linear growth of the main axis. With IAA decreasein number of laterals closely parallels the decrease in lineargrowth of the main axis; with IAN reduction in linear growthof the main axis occurs at concentrations above 10^–8 whereasnumber of laterals does not decrease until the concentrationexceeds 10^–6.
2. Study of the course of cell elongationin the exodermal cellsshowed that in the standard medium andin media containing 510^–9IAA or 510^–6 IAN theprocess takes about 7 hours; thefinal cell lengths in IAA andIAN media are lower than in standardmedium owing to a slowerrate of elongation. The decrease inlinear growth of the mainaxis in presence of IAA could be accountedfor by the decreasein cell length; this was not the case withIAN. The implicationsof this are considered.
3. Determinations of the distance (mm.)between, and of the numberof exodermal cells separating, theadjacent laterals in oneorthostichy showed that IAN enhancesthe frequency of lateralswhereas this is either unaffectedor decreased by IAA. The enhancementof lateral frequency inIAN arises from shortening of the cellsof the main axis anddecrease in the number of cells separatingadjacent laterals.
4. The results are considered to support the view that IAN haseffects on root growth different from those of IAA. Study ofthe degree of inhibition of main axis growth and of alterationsin lateral frequency resulting from treatment with mixturesof IAA and IAN provided data which could also be most easilyexplained on this hypothesis.

     

The Carbohydrate Nutrition of Tomato Roots: III. The Effects of. External Sucrose Concentration on the Growth and Anatomy of Excised Roots

Variation of the concentration of sucrose in White‘s mediummarkedly affects the increase in length of the main axis, andthe number and total length of the laterals developed by excisedtomato roots. Highest values for these three features are obtainedwith medium containing 1.5 per cent, sucrose. Root materialhas been analysed anatomically. Root diameter, cortical thickness,number of xylem vessels, and development of the cortical air-spacesystem increase with increase in sucrose concentration. Thedegree of vacuolation of extra-stelar meristematic cells increases,and protophloem differentiates further from the promeristemwith rising sucrose concentrations. The interpretation of theseresults is discussed.     

Metabolism of Some Indole Auxins in Excised Tomato Roots

Indolylacetylaspartic acid (IAAsp) and possibly indolylacetylglutamicacid (IAG) are formed by exposure of excised tomato roots toIAA. Little ‘free’ IAA accumulates in the tissue.An unidentified substance reacting pink with nitricnitrite reagentis also formed. These substances are metabolized when IAA-treatedroots are transferred to auxin-free medium. IAAsp and IAA aresimilarly inhibitory to the growth of excised tomato roots.Excised tomato roots do not interconvert IAA and IAN. IAN-feedingleads to IAN accumulation and the appearance of indolylcarboxylicacid (ICA); transference to auxin-free medium causes a declinein the IAN activity but the ICA spot persists. The inhibitoryactivity of IAN is not due to its conversion to ICA. Excisedtomato and wheat roots respond very differently to externall-tryptophane but in neither case is there evidence of the conversionof tryptophane to ethyl acetate-soluble auxins.     

Sugar Absorption and Sucrose Inversion by Excised Tomato Roots

The growth of excised tomato roots in sucrose is accompaniedby the appearance in the medium of glucose and fructose. Thequantitative relations between this appearance of glucose andfructose in the medium, total sugar absorption by the roots,and the decrease in the sucrose concentration of the mediumdo not suggest any causal relationship between sucrose uptakeand glucose and fructose appearance in the medium. Excised tomato roots exhibit surface invertase activity witha pH optimum at 4.0–4.4. Alterations of external pH, whichdid not affect sucrose absorption, drastically altered the levelsof glucose and fructose appearing in the medium. Glucose is preferentially absorbed from mixtures of glucoseand fructose, and by adjustment of the ratio of the two sugars,a mixture can be obtained from which equimolar absorption ofthe two sugars occurs. Root growth in this mixture is, however,very poor compared with that occurring in presence of sucrose. The results are discussed in the light of earlier studies onsucrose uptake by cultured tomato roots.     

Effects of Growth Retardants on the Growth of Excised Roots of Dolichos lablab L. in Culture

The effects of various growth-retarding chemicals on the growthof excised roots of Dolichos lablab in sterile culture are investigated.Application of a range of concentrations of CCC, phosfon, andB-995 inhibited growth in length of the roots by reducing thefrequency of cell division, with little or no effect on cellelongation. Treated roots had generally lower DNA, RNA, andprotein contents than controls, although CCC-treatment significantlyenhanced the soluble nitrogen content of the roots. The inhibitoryeffects of growth retardants were not reversed by IAA or GA,but substances like choline chloride and pyridoxine hydrochloridewere partially effective in reinstating normal growth in rootsgrowing in concentrations of CCC producing about 50 percentinhibition of growth in length.     

Gibberellins and the Growth of Excised Tomato Roots: Comparison of gib-1 Mutant and Wild Type and Responses to Applied GA3 and 2S, 3S Paclobutrazol

Clones of excised roots of wild type tomato (Lycopersicon esculentum,Mill., cv. Moneymaker) and a near-isogenic GA-deficient mutant(gib-1/gib-1) were cultured in modified White's medium containing1.5% w/v sucrose. The linear elongation rate of the main axisof the gib-1 mutant was 40% less than that of the wild type.In addition, the main axis of the gib-1 mutant was thicker thanthat of the wild type but main axis volume growth was the samein both genotypes, indicating that the gib-1 allele was affectingthe orientation of root expansion. There was no evidence tosuggest that the gib-1 allele affected either the pattern ofemergence or the density of lateral roots. Elongation rate andthickness of gib-1 mutant roots were restored to those of thewild type by the addition of low concentrations (0.1–1.0µM) of gibberellic acid (GA3). These concentrations ofGA3 caused a slight reduction in extension growth of wild typeroots, indicating that endogenous GAs were not limiting elongationof normal roots in culture. The GA biosynthesis inhibitor, 2S,3S paclobutrazol, at 0.1 µM, significantly reduced elongationof wild type roots and this inhibition was counteracted by 0.1µM GA₃. It is concluded that the difference in growthbetween the gib-1 mutant and the wild type represented GA-dependentgrowth. Low concentrations of 2S, 3S paclobutrazol caused onlya small (5%) reduction in growth of the gib-1 mutant and thisgrowth inhibition was not reversed by GA₃. This observation,and the fact that gib-1 mutant roots grow in the absence ofadded GA₃, suggested that part of root growth was GA-independent.However, the possibilities that the gib-1 mutant is ‘leaky’and that paclobutrazol does not inhibit GA biosynthesis completelycannot be excluded. Key words: gib-1 mutant, gibberellic acid, Lycopersicon esculentum, 2S, 3S paclobutrazol, root growth     

Studies of the Growth in Culture of Excised Wheat Roots

Evidence is presented that the growth of cultured wheat roots is enhanced in closed culture systems not only by carbon dioxide but by a further and apparently very volatile product of root metabolism. The formation of this second growth-stimulating substance is not considered to depend upon a reduction of oxygen tension in the culture system. Unlike ethanol the activity of the volatile promoter does not depend upon illumination of the cultures. The volatile promoter cannot be effectively absorbed by a number of gas absorbents. Attempts to demonstrate ethylene production by the roots have been unsuccessful.     

Studies of the Growth in Culture of Excised Wheat Roots

Aeration of root culture medium with air containing 5 % or 10 % CO₂ enhances growth, particularly lateral initiation and growth, compared with aeration wilh air or CO₂-free air. This beneficial effect of enhanced CO₂ pressure on the growth of cultured roots of Atson Elite 56 wheat ean also be demonstrated in presence of a high (40 %) and inhibiting level of oxygen. ¹⁴C labelling from a carbonate–bicarbonate solution of pH 7.0 by cultured wheat roots is enhanced by light and this labelling is not inhibited by the presence of 1 % ethyl urethane.     

Effects of Exudation on Ion Relationships of Excised Roots

In excised tomato roots submerged in solution, mannitol at –2.8 atm increased the phosphorus uptake and decreased the loss of previously absorbed phosphorus. Separate collection of the xylem exudate demonstrated that these mannitol effects were due entirely to a reduced phosphorus flow via exudation. For example, in the case of previously absorbed phosphorus, the high loss of phosphorus at –0.3 atm could be contributed to phosphorus transport via the exudation stream, which was higher at –0.3 than at –2.8 atm. In contrast, loss from the root surface to the external medium was identical for the two different water potentials. The neglect in measuring ion flow in the exudate might have confounded ion transport studies by other workers. Some particular cases were re-examined, such as chloride uptake at high external concentrations and ion toss from different cell compartments.     

Studies of the Growth in Culture of Excised Wheat Roots

Excised wheat roots in culture release a substance inhibitory to their growth and which can be absorbed on activated charcoal. This growth inhibitor can be isolated in pure form by diethylether extraction followed by DEAE cellulose and thin-layer chromatography. At very low concentration it inhibits lateral root development. The significance of this inhibitor to the problem of the continuous growth of cultured wheat roots is discussed.