Amino acids liberated into the culture medium by pea seedling roots

Summary The amino acid composition of the medium in which seedlings of peas had been grown aseptically with and without quartz sand were determined by ion-exchange chromatography. Besides considerable amounts of amino acids, peptide material was also present.     

Amino acids in root exudates of cotton

Summary Cotton-field soils from a wilt-free zone (KPT) and from a wilt-sick zone (PLD), and root exudates of diploid and amphidiploid cotton strains grown in KPT and PLD soils were analyzed for 13 free amino acids employing microbiological assay techniques. Test organisms used wereLactobacillus arabinosus 17/5 andLeuconostoc mesenteroides P. 60.Arginine, proline, lysine, serine, glycine, cystine, and tyrosine occurred in both the soils; amounts of these were higher in PLD soil than in KPT soil.Seven to eleven amino acids were found in the root exudates of cotton. Aspartic acid, threonine (in one), phenylalanine and leucine were found in root exudates but not in the control soils. Diploid strains were found to exude greater amounts of amino acids than the amphidiploid strains. Comparing the effect of the two soils tested, it was found that amino acids in exudates of plants grown in PLD soil were higher except in the case of one diploid strain.In the inoculated series, amino acids in the exudates were markedly lower than the respective healthy controls in the diploid strains. In the exudates of amphidiploid strains also, amino acid contents were reduced but to a lesser extent.Significance of occurrence of cystine in the exudates of some strains is indicated.Part of Doctoral thesis, University of Madras.     

Flavonoids from shoots, roots and roots exudates of Brassica alba

Analysis of extracts obtained from shoots, roots and exudates of Brassica alba revealed the presence of 3,5,6,7,8-pentahydroxy-4'-methoxy flavone in shoots, as well as 2',3',4',5',6'-pentahydroxy chalcone and 3,5,6,7,8-pentahydroxy flavone in roots and exudates. Apigenin was also found in the shoots and roots, but not in the root exudates.     

Extracellular proteins in pea root tip and border cell exudates

Newly generated plant tissue is inherently sensitive to infection. Yet, when pea (Pisum sativum) roots are inoculated with the pea pathogen, Nectria haematococca, most newly generated root tips remain uninfected even though most roots develop lesions just behind the tip in the region of elongation. The resistance mechanism is unknown but is correlated spatially with the presence of border cells on the cap periphery. Previously, an array of >100 extracellular proteins was found to be released while border cell separation proceeds. Here we report that protein secretion from pea root caps is induced in correlation with border cell separation. When this root cap secretome was proteolytically degraded during inoculation of pea roots with N. haematococca, the percentage of infected root tips increased from 4% +/- 3% to 100%. In control experiments, protease treatment of conidia or roots had no effect on growth and development of the fungus or the plant. A complex of >100 extracellular proteins was confirmed, by multidimensional protein identification technology, to comprise the root cap secretome. In addition to defense-related and signaling enzymes known to be present in the plant apoplast were ribosomal proteins, 14-3-3 proteins, and others typically associated with intracellular localization but recently shown to be extracellular components of microbial biofilms. We conclude that the root cap, long known to release a high molecular weight polysaccharide mucilage and thousands of living cells into the incipient rhizosphere, also secretes a complex mixture of proteins that appear to function in protection of the root tip from infection.     

Cytokinins in seedling roots of pea

The natural occurrence of cytokinins existing both in a free form and as a constituent of transfer RNA was examined in serial segments of young seedling roots of pea. Purified ethanol extracts of root apices were resolved into four factors capable of inducing soybean callus tissue proliferation. The most active factor was identified as zeatin or some closely related compound; it produced polyploid divisions and tracheary element differentiation when tested on cultured pea root segments. The terminal 0- to 1-millimeter root tip contained 43 to 44 times more free cytokinin on a fresh weight or a per cell basis than the next 1- to 5-millimeter root segment. Extracts of more proximal segments behind the tip contained no measurable free cytokinin. Acid hydrolysates of transfer RNA exhibited reproducible cytokinin activity. Bioassays revealed that the predominant amounts of free cytokinin and that present in transfer RNA were restricted to the extreme root tip. There was approximately 27 times more free cytokinin than the amount detected in transfer RNA in root apices.     

In vitro loss of nucleic acids from cells of aseptically cultured excised pea roots

Summary Analysis of pea root tips taken from attached seedling roots and excised roots cultured in vitro has revealed major differences in cell constituents. The cells of cultured roots have only 40% and 13% of the protein and amino acid content of attached root cells. The nucleic acid content of cultured root cells was shown to be only 20% and 27% of the RNA and DNA respectively found in attached roots. It is suggested that there is excess nucleic acid in whole plant tissues above that required for transfer of genetic information necessary for normal growth and differentiation of root cells.     

Phenolic acids in groundnut seed exudates

Summary Seed exudates were collected aseptically and extracted for phenolic acids. Paper chromatographic analysis revealed the presence of eleven phenolic compounds.Department of Botany, S.V. University, Tirupati-517502, Andhra Pradesh, India.     

Aldrin epoxidase from pea roots

The membrane system which epoxidizes aldrin to dieldrin in pea roots was separated from polyphenolic inhibitors and soluble peroxidase by rapid anaerobic gel filtration. Epoxidase activity was strongly inhibited by cytochrome-c, cyanide, electron acceptors and polyphenols and was enhanced by chelating and solubilizing agents which inhibit cytochrome P450 systems.     

Regeneration of vascular tissue in wounded pea roots

Severance of the stele of young main roots of pea (Pisum sativum L.) results in formation of a bridge of vascular tissue in the remaining cortex. Cell divisions occur close to the severed vascular tissues on both the proximal and distal sides of the cut within 24 h. Differentiation of new vascular strands subsequently begins in the same locations and progresses from both sides of the wound into the remaining cortex and also back along the original vascular strands. Most of the vascular tissue which forms the bridge through the cortex differentiates in the acropetal direction. Continuous strands composed of single sieve elements bypass the wound somewhat sooner than the first complete xylem strands; the latter in 60–70% of the cases, are present by 3 d. Cambial activity subsequently adds more xylem and phloem. Vascular regeneration is not affected by removal of the epicotyl or the root tip; it is greatly reduced but not prevented by removal of the cotyledons.     

Amino acids response of glutamate dehydrogenase from light and dark treated roots and shoots of maize

Thein vitro activity of glutamate dehydrogenase (NADH-GDH), from dark-treated root segments of maize seedlings responded differently to amino acids threonine, glutamate and methionine than that from light-treated root segments, and to the amino acid methionine in dark- and light-treated shoot segments. In most cases amino acids inhibited GDH activity, the inhibition increased with amino acid concentration. However, methionine activated GDH from dark-treated roots and light-treated shoots, while aspartate had little effect on enzyme activity.     

Rapid modulation of nitrate reductase in pea roots

The regulatory properties of nitrate reductase (NR; EC 1.6.6.1) in root extracts from hydroponically grown pea (Pisum sativum L. cv. Kleine Rheinländerin) plants were examined and compared with known properties of NR from spinach and pea leaves. Nitrate-reductase activity (NRA) extracted from pea roots decreased slowly when plants were kept in the dark, or when illuminated plants were detopped, with a half-time of about 4 h (= slow modulation in vivo). In contrast, the half-time for the dark-inactivation of NR from pea leaves was only 10 min. However, when root tip segments were transferred from aerobic to anaerobic conditions or vice versa, changes in NRA were as rapid as in leaves (= rapid modulation in vivo). Nitrate-reductase activity was low when extracted from roots kept in solutions flushed with air or pure oxygen, and high in nitrogen. Okadaic acid, a specific inhibitor of type-1 and type-2A protein phosphatases, totally prevented the in vivo activation by anaerobiosis of NR, indicating that rapid activation of root NR involved protein dephosphorylation. Under aerobic conditions, the low NRA in roots was also rapidly increased by incubating the roots with either uncouplers or mannose. Under these conditions, and also under anaerobiosis, ATP levels in roots were much lower than in aerated control roots. Thus, whenever ATP levels in roots were artificially decreased, NRA increased rapidly. The highly active NR extracted from anaerobic roots could be partially inactivated in vitro by preincubation of desalted root extracts with MgATP (2 mM), with a half-time of about 20 min. It was reactivated by subsequently incubating the extracts with excess AMP (2 mM). Thus, pea root NR shares many of the previously described properties of NR from spinach leaves, suggesting that the root enzyme, like the leaf enzyme, can be rapidly modulated, probably by reversible protein phosphorylation/ dephosphorylation.     

Relationships between nodulation and auxin level in pea roots

In the roots of 4-day-old pea plants germinated in unsterile soil from Rhizobium-inoculated seeds, a higher level of native IAA was determined than in roots of pea plants germinated in sterile soil from superficially sterilized seeds. The IAA level in plants grown from inoculated seeds increased markedly up to the age of 6 days of the plant, while in plants growing under sterile conditions it did not significantly change during the same period. Between the 6th and 10th days of the age of the plant, a decline in the IAA level was observed in roots of plante growing from inoculated seeds. It was not until after 10 days of age of the plant that the level of IAA in nodulated roots again increased.     

Membranes carrying acid hydrolases in pea seedling roots

Subcellular localization of acid hydrolases in pea seedlingroots was studied by differential and sucrose density gradientcentrifugations. Significant parts of hydrolase activities inthe tissue were recovered in mitochondrial and microsomal fractions.Sedimentable phosphatase was separated into two subtractions:denser and lighter membrane fractions. The distribution of phosphataseactivity after sucrose density gradient centrifugation of thedenser fraction coincided with that of antimycin AinsensitiveNADH-cytochrome c reductase activity. Electron microscopic observationssuggested that the fraction contained only microsomes. RNasein the denser fraction seemed to associate with ribosomes. Phosphataseand RNase were solubilized by sonic treatment in the presenceof high concentrations of salt. On the other hand, a-amylasewas tightly bound to a membrane. The results are discussed withspecial regard to the relationship between the membranes andlysosomes. (Received May 4, 1973; )     

Effect of ammonia on sucrose synthase in pea roots

The effects of ammonium on activity of sucrose synthase (SS) in the roots of pea (Pisum sativum L.) plants were studied. On the medium containing 14.2 mM (NH₄)₂SO₄, SS activity increased by 20–200% for 10–20 days of plant growth as compared with the roots of plants growing without nitrogen. Illuminance affected the degree of effects. Under natural illumination, ammonium affected SS activity not only in sunny days (up to 25 klx) but also in cloudy days (3–6 klx) but to a lower degree. Under stable low light (2.5 klx), ammonium did not affect SS activity. In the in vitro experiments, at (NH₄)₂SO₄ concentrations from 0 to 1 mM, SS activity was suppressed (up to 10%), whereas 1–37.5 mM (NH₄)₂SO₄, it was increased (up to 50%).