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

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.     

The Nutrition of Excised Wheat Roots

From an investigation of the nutrient requirements of Hilgendorf wheat roots, glucose emerged as the only suitable source of carbon and energy, and thiamine as the only vitamin necessary. The roots had the ability to utilise nitrogen from calcium nitrate, urea, alanine, glutamine, allantoin, and hydrolyzed casein.     

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.     

The Effects of Gibberellins on the Growth of Excised Tomato Roots

1. At appropriate concentrations both gibberellic acid (GA) and1-naphthalene-acetic acid (NAA) enhance the main axis growthof excised tomato roots grown in culture media containing sucroseat concentrations below 1 per cent. Lateral root extension growthis enhanced by GA at all sucrose concentrations tested; onlyat the lower sucrose concentrations is this effect observedwith NAA. Both GA and NAA increase the number of emergent lateralroots and this effect is most marked in media of low sucrosecontent. Both GA and NAA at higher concentrations inhibit rootgrowth but NAA exhibits its full range of growth effects overa much narrower concentration range than GA.
2. GA, like NAA,speeds up the loss of meristematic activity whichoccurs whenindividual meristems are repeatedly subculturedin media containing1 per cent, or higher concentrations ofsucrose.
3. The promotionof main axis growth by both GA and NAA involvesenhanced cellelongation and cell division. At a moderatelyinhibitory concentrationGA reduces both cell elongation andcell division; this is notthe case with NAA.
4. Gibberellins A₁, A₂, and A₄ resemble GA(gibberellin A₃) intheir growth effects. Allogibberic acidlike G A promotes lateralroot extension growth but causes markedinhibition of root growthat a much lower concentration thanGA.

     

Sterile Culture of Excised Tomato Roots in Sands of Different Grain Size

A simple technique to grow excised tomato roots in sand in sterileculture is described. The apparatus consists of a culture tubefilled with sand linked to a small flask containing the nutrientsolution. By raising the lower-placed flask the sand is flushedwith solution, thereby replenishing nutrients at the root surface.Lowering the flask again allows air to enter the sand, thuseffecting sufficient aeration. It is further demonstrated that growth of the roots is relatedto acration and not to mechanical impedance in sands of differentgrain size.     

Growth-Promoting Effects of Fatty Acids on Excised Wheat Roots and Oat Coleoptiles

The growth action of some fatty acids and alcohols with carbon number from 1 to 4 was tested on excised wheat roots in aseptic cultures. Growth, cell length, and dry weight were measured after seven days. The tested substances were: Formic acid, acetic acid, propionic acid, n-Butyric acid, isobutyric acid, acelaldehyde, propionaldohyde, ethanol, n-propanol, isopropanol, n-butanoL and isobutanol. The primary alcohols and all of the acids, except formic acid, promoted the cell elongation and the total growth, but the meristematic activity was inhibited in the higher concentrations. The concentrations with maximal growth-promoting activity were 10^?2M to 10^?3M for the alcohols and 10^?4M for the acids. Propionic acid was applied in darkness, red light, and white light and proved to increase the cell length from 215 μ, 180 μ, and 120 μ up to 240 μ in all three treatments. The growth rate was not affected, but the duration of the cell elongation was extended. The presence of iron proved to be necessary for the stimulation. The chlorophyll content in the light grown roots was relatively unaffected when the cell length was increased and the dry weight was not increased as long the cell number was normal. The growth of Avena coleoptile segments was slightly promoted by propionic acid in the presence of IAA.     

Potassium Fluxes in Excised Barley Roots

The method of the modified compartmental analysis for excisedroots has been adopted for measuring K⁺-fluxes and compartmentationin barley (Hordeum distichon) roots. Efflux of ⁴²K and ⁸⁶Rbindicated that more than two intracellular compartments wereinvolved in the tracer exchange; the ⁴²K data clearly showedthe components. On the basis of the efflux behaviour of theapical and more basal tissues of the roots, the three componentsof efflux were attributed to the cytoplasm of differentiated(fast) and meristematic tissues (intermediate) and to the vacuoles(slow exchange) of the roots. A model is proposed on the basisof which, the fluxes corresponding to the meristematic and differentiatedtissues of the root can be estimated. Additionally, fluxes ofthe differentiated root tissues were determined by using effluxdata obtained with root segments without apical tissues. Thedata obtained in both ways compare reasonably well and agreeto independent chemical measurements. Comparison of the ⁴²K and ⁸⁶Rb efflux data show strong discriminationof K^– in favour of Rb⁺ and indicate that ⁸⁶Rb is not suitableas a tracer for K⁺ in efflux measurements, at least with barleyroots.     

Changes Induced by Salinity to the Anatomy and Morphology of Excised Pea Roots in Culture

Ann. Bot. 57, 811–818, 1986     

Changes Induced by Salinity to the Anatomy and Morphology of Excised Pea Roots in Culture

Excised pea roots were grown in culture in the absence or presenceof NaCl. Salinity induced anatomical and morphological changesin the roots, some of which could be observed after only 24h in culture. Roots became constricted just above the apex,the region above the constriction thickened and the root tipcurved through 90°. Cellular differentiation began nearerthe apex, cortical and epidermal cells shortened and mitoticactivity in the pericycle increased as a result of exposureto salinity. Some of the changes resemble those induced by ethylene,but ethylene probably was not the cause of the response to salinity.Root cultures seem to be a suitable model for studying the effectof salinity in plant roots. Pisum sativum cv. Alaska, salinity, roots, cortex, root culture, pericycle, growth and differentiation     

Salt-induced Changes in the Distribution of Amyloplasts in the Root Cap of Excised Pea Roots in Culture

The effects of 120 mM NaCl on the anatomy and ultrastructureof the root tip of cultured excised pea roots was investigatedafter 24 h exposure to salinity. In the meristematic cells mitochondrialdamage was apparent and these cells showed increased vacuolation.The root cap was already severely affected after 24 h exposureto salinity and clumping of the cap amyloplasts around the cellnuclei was apparent. The possibility that salinity may affectroot gravitropic responses is discussed. Pisum sativum L. cv. Alaska, salinity, roots, root culture, amyloplasts, ultrastructure     

Stimulation of the Growth of Excised Cultured Roots of Soya Bean by Abscisic Acid

Excised root cultures of soya bean cultured at 30 °C inWhite's medium (1943) supplemented with 0•1 per cent yeastextract have been serially sub-cultured over 13 culture passagesof 7 days although a decline in the linear growth and lateralformation begins in the third passage and the roots are devoidof laterals from the 8th passage onwards. Applications of abscisicacid within the concentration range 0•01–0•001mg l^–1 and during the first two culture passages enhancedthe linear growth of the main axis, the number of emergent lateralsand the total length of laterals. This effect has been shownfor two cultivars, with roots derived from seed in both itsfirst and second year of storage and under different conditionsof culture and culture pH.     

Some Effects of Light on Excised Wheat Roots with Special Reference to Peroxide Metabolism

When excised wheat roots are exposed to blue light, catalase activity changes in a way to he expected as a result of photodestruction with first order kinetics. Wheat root catalase in vivo is less light sensitive than animal catalase in vitro, possibly due to internal screening. Illumination of the roots with red light causes some increase in catalasc activity. Peroxidase activity is much less affected by light. No relation has been found between catalase destruction and chlorophyll formation. The ability of roots to oxidize pyrogallol to purpurogallin (by other workers interpreted as peroxide production) is decreased by light, especially blue light. On the contrary, one peroxide producing enzyme, glycolic acid oxidase, was detected only in roots grown in blue light. The total flavin content, or the fraction present as FAD, is not affected by light. The ascorbic acid content is low, but slightly increased by blue light.