Source of Induction and Sites of Primary Phloem Fibre Differentiation in Coleus blumei

The fact that fibre induction is strictly basipetal is usedhere to study the long distance effect of young growing leaveson acropetal primary phloem fibre differentiation. Excisionexperiments are used to show that young leaves induce fibredifferentiation around a wound a few internodes below them.No fibers appeared in the younger internodes between the youngleaves and the mature internode. Young leaves yield shorterfibres than those which differentiate under mature leaves, indicatingthat more than one stimulus is involved in the induction process.Fibre differentiation in nodes is faster than in subtendinginternodes. Wounding causes rapid differentiation of phloemfibres above and beside the wound. The rapid differentiationin the node as well as around the wound can be understood asan effect of a high local concentration of inductive stimulus.It is proved that the ability of the cells to respond to inductiondetermines the pattern of their differentiation which in thiscase is counter-directional to the induction. Coleus blumei, phloem fibres, differentiation     

Phloem Unloading within the Seed coat of Pisum sativum Observed using Surgically Modified Seeds

Phloem unloading in pea seed coats was observed by removingthe embryos from developing seeds and washing the attached coatswith a weakly buffered solution. The quantity of labelled photosynthateappearing in the washing solution varied immediately when thesolute concentration was changed, and is shown to be an osmoticresponse. This response is predicted by the Münch theoryof phloem transport with concentration dependent unloading.Respiratory inhibitors and the sulphydryl modifying reagentPCMBS had a slow effect upon the washout of tracer, which arrivedwithin the seed coat prior to inhibitor application, but completelystopped any washout of tracer arriving after its application.This time-course suggests that the inhibitors were not directlyinhibiting unloading, but preventing further tracer from enteringthe region of unloading within the seed coat. Phloem unloadingwithin the seed coats of Pisum appears to be passive and notdependent upon a PCMBS-sensitive carrier. Key words: Pisum sativum, seeds, phloem unloading     

Stomata on the Primary Root of Pisum sativum L.

The first record of stomata on a non-specialized root was obtainedby scanning electron microscopy of 4-d-old Pisum sativum L.In some cases subsidiary cells were trichoblasts. Stomata andthe root triarch vascular structure were simultaneously presentin transverse sections through the root. Pisum sativum, pea, root stomata, guard cells, trichoblasts     

Lipoxygenase heterogeneity in Pisum sativum

Antibodies raised against two pea (Pisum sativum L. cv. Birte) seed lipoxygenases have been used to analyze lipoxygenase heterogeneity in seeds and in other organs. At least seven different polypeptides were identified in vivo; five of these were identified as precursors synthesized in vitro. The developmental appearance of the seed polypeptides has been analyzed and early and late forms were identified. Limited N-terminal sequence data indicated further heterogeneity when compared with sequences predicted from cDNAs.Abbreviations cDNA complementary DNA - DAF days after flowering - HPLC high-performance liquid chromatography - Ig immunoglobulin - kb kilobase - M_r relative molecular mass - PAGE polyacrylamide gel electrophoresis - PVDF polyvinylidene difluoride - SDS sodium dodecyl sulphate - SSC 0.15 M sodium chloride, 0.015 M sodium citrate, pH 7.0 This work was supported by the Agricultural and Food Research Council via a grant-in-aid to the John Innes Institute. We acknowledge financial support from the Commission of the European Communities Biotechnology Action Programme; grant No. 0063-UK.     

Phloem exudate metabolic content reflects the response to water-deficit stress in pea plants (Pisum sativum L.)

Drought stress impacts the quality and yield of Pisum sativum. Here, we show how short periods of limited water availability during the vegetative stage of pea alters phloem sap content and how these changes are connected to strategies used by plants to cope with water deficit. We have investigated the metabolic content of phloem sap exudates and explored how this reflects P. sativum physiological and developmental responses to drought. Our data show that drought is accompanied by phloem-mediated redirection of the components that are necessary for cellular respiration and the proper maintenance of carbon/nitrogen balance during stress. The metabolic content of phloem sap reveals a shift from anabolic to catabolic processes as well as the developmental plasticity of P. sativum plants subjected to drought. Our study underlines the importance of phloem-mediated transport for plant adaptation to unfavourable environmental conditions. We also show that phloem exudate analysis can be used as a useful proxy to study stress responses in plants. We propose that the decrease in oleic acid content within phloem sap could be considered as a potential marker of early signalling events mediating drought response.     

The Isoperoxidases of Pisum sativum

The heterogeneity of the peroxidases in peas was examined by starch gel electrophoresis. Comparisons were made between tall and dwarf cultivars and among organ systems developed in light and darkness. Isoperoxidase bands could be grouped as cathodic, anodic and near-neutral (at pH 9.0) types. The cathodic set stained well with guaiacol oxidation products whereas some anodic bands reacted preferentially with 2,6-dimethoxyphenol. Some near-neutral bands were aceto-carmine positive and may have been organellar.Each organ had a characteristic isozyme pattern, and the band patterns in corresponding organs from different varieties were far more alike than were the patterns in the different organs within each variety. Ontogenetic changes were marked in all 3 organ systems, principally in the cathodic bands. The effect of light on isozymal patterns was quantitative rather than qualitative, possibly influencing the isoperoxidases secondarily via its effect upon organ physiology and development.     

Matromorphy in Pisum sativum L.

Summary The possibility of obtaining instant pure breeding lines by matromorph seed development in Pisum sativum L. has been investigated. Two types of maternal parents, namely, homozygous for the recessive marker genes and heterozygous for the dominant marker genes were pollinated with Lathyrus odoratus and the P174 variety of Pisum sativum L. carrying dominant markers. For both pollinators, induction of matromorphy by prickle pollination, irradiated pollen and IAA treatment was examined. Promising matromorphs were identified in the M₁ generation which were studied in the M₂ generation for assessing their genetic status with respect to homozygosis. The success of pod set varied from zero to 28% with a varying number of matromorphic seeds following different treatments. The possible mechanisms for matromorphic origin have been discussed. The evidence presented herein favours induction of matromorphy in peas for the production of homozygous stocks. In addition, the recovery of double recessive seed markers of the maternal parents along with plant markers from the paternals has prospective implications in plant breeding as an alternative tool to recurrent back crossing.     

Phytochrome-mediated bud development in Pisum sativum

Summary Light-grown dwarf peas were disbudded except for a single lateral bud, then transferred to darkness at 24° C. During the dark period the seedlings were irradiated daily for 5 or 7 min with R or FR. The buds exposed to R developed into shoots faster than those irradiated with FR. The R effect was FR reversible, and the FR effect was R reversible. The P_fr form of phytochrome thus promoted shoot growth including cell division, DNA and RNA synthesis.Abbreviations R red - FR far-red - P phytochrome     

Cell Division Heterogeneity in the Primary Meristems of Pisum sativum Root Tips after Excision

Root tips (1.0 cm) were excised, transferred to culture medium,sampled at intervals, embedded in paraffin and sectioned. Themitotic index in each primary meristem in the 2.0 mm tips wasreduced immediately after excision. After 8–12 h mitosisresumed. The position of mitotic figures in the protoderm, groundmeristem, procambium, and root cap was determined. The greatestnumber of mitotic figures occurred in the procambium, and uponexcision the smallest percent reduction also occurred in thisregion. The ground meristem showed the greatest reduction inmitotic figures. After excision, the relative contribution tothe total number of mitotic figures increased in the procambiumtissue and decreased in the ground meristem. Cell division inall primary meristems of the root tip is reduced after tip excision,but the ground meristem (cortical region) is the primary targettissue.     

Auxin Physiology of Dwarfism in Pisum sativum

Similar levels of diffusible auxin are measured for the apices of both Little Marvel (dwarf) and Alaska (normal) cultivars of the pea when grown in sunlight and darkness. In sunlight, however, diffusible auxin disappears in the subtending internode of the Little Marvel plant but remains at 50 per cent of the level of the apex in the subtending internode of the Alaska plant. The enzyme preparation from the apex of the dwarf plant converts tryptophan and tryptamine to IAA more readily than that from the normal plant. Indoleacetyl aspartate synthetase activity is also higher in the dwarf plant than in the normal plant and the dwarf plant contains four times as much conjugate as the normal plant with or without treatment with gibberellic acid. Gibberellic acid (GA) does not affect the induction of the synthetase enzyme nor the enzymatic formation of indoleacetyl aspartate. The growth induced by GA is the result of an increased synthesis of auxin.     

Peculiarities in PMC Meiosis of Pisum sativum

Summary Nemesia strumosa plants were discovered which had styles capable of discriminating among incompatible pollen tubes from different pollinators, allowing growth of some but not others. All but 3 of 26 families tested had at least some members with discriminating styles (DS). Presence and level of DS was independent of S genotype. Plants with pseudo-self-compatiblity (PSC) levels greater than 10% had the trait, though many plants with strong DS had PSC levels less than 10%. Self pollination of highly DS plants produced mostly DS offspring, but of differing sensitivities. Some progenies from crosses between a family of highly DS plants and unrelated, probably low DS plants segregated half DS and half non-DS, while others consisted of mostly DS or mostly non-DS. The DS phenomenon is probably caused by PSC genes.Scientific Journal Series Paper No. 11,677 of the Minnesota Agricultural Experiment Station     

Glucolipids of Zea mays and Pisum sativum

The glucolipids formed upon feeding (U-¹⁴C)glucose to embryos of Zea mays were partially characterized with respect to: (a) metabolic turnover, (b) acid lability, (c) phosphorus content, (d) chromatographic properties, and (e) hydrolysis products. The chloroform-methanol-soluble assimilated radioactivity was examined specifically for occurrence of a glycosylated prenol phosphate. With the extraction conditions used, no evidence was found for formation of a glucosylated prenol phosphate. Several, as yet unidentified, acid-labile glucolipids undergoing metabolic turnover were observed. Four diglycerides were characterized as hydrolysis products of a fraction that contained ¹⁴C-glucose and phosphorus, and was subject to metabolic turnover. Examination of the 1-butanol-soluble glucolipids from pea (Pisum sativum) seedlings also demonstrated anionic glucolipids, evidencing metabolic turnover but none with the properties of glucosylated prenol phosphate.     

Interrelationships of Cotyledonary Shoots in Pisum sativum

Unequal two shoot systems of tall and dwarf varieties of Pisum sativum L. ev, Improved Pilot and Meteor, respectively, were used in experiments where manipulative treatments involving the removal of one or other or both of the shoot apices were carried out. These were combined with GA treatments. Twenty four hours after treatment, ¹⁴CO₂ was applied to the lowest expanded leaf on the larger shoot, and the distribution of radioactivity was determined after a further 24 hours. Removal ot the apex of the larger shoot reduced the level of translocation from the treated leaf, the effect being enhanced by the additional removal of the other apex. This (Heel was more marked in the tall than in the dwarf variety. In untreated control plants of both varieties there was very little transfer ot labelled material from the larger (dominant) shoot to the smaller (weaker) shoot. This pattern was not affected by the removal of the apex from the weaker shoot. In Improved Pilot, removal of the apex from the stronger shoot led to considerable transfer of radioactive material to the weaker shoot, GA treatment having little effect. However, in Meteor, transfer of radioactivity from the stronger to the weaker shoot. after removal of the apex from the stronger shoot, only occurred after the application of GA. Removal of both shoot apices again resulted in low levels of transfer of labelled material to the weaker shoot in both varieties. These results are consistent with the hypothesis that there is competition between the two shoots for photosynthates produced by the leaves and that treatments reducing the competitive ability of one shoot, reduce the level of nutrients received by that shoot and can result in transfer of materials from it to the other shoot. Continuation of such a situation will result in increased inequality and perhaps ultimately in the death of the weaker shoot.     

Genome size variation in Pisum sativum.

Pisum sativum L. is one of the plant species where infraspecific genome size variation, up to 1.29-fold between cultivars, has been reported. The present investigation deals with a Feulgen cytophotometric analysis of this phenomenon in 25 wild accessions, landraces, and cultivars of widely different geographic origin. Differences between accessions were maximally 1.054-fold in single experiments but proved to be nonreproducible upon repeated measurements. Seedlings of the same accession often differed significantly, up to 1.056-fold, but values from root and shoot tips in one individual were not significantly correlated, indicating the absence of true genome size variation between plants. Upon calibration against Allium cepa a 1C value of 4.42 pg is estimated for Pisum sativum. Altogether the data suggest that, contrary to the divergence in the literature data and recent reports on DNA content variation, the pea has a stable genome size.     

Gibberellin translocation in Pisum sativum L.

The physiological and biochemical consequences of treating Le (tall) and le (dwarf) pea seedlings with varying quantities of the gibberellins [³H]GA₂₀ and GA₁ have been investigated. Although the percentage uptake of these compounds from the site of application on the 3 stipules was low and most of the applied GA remained unmetabolised in situ, the quantitative relationship between GA translocation and GA dosage was found to be linear for GA₁ but saturating for GA₂₀. The movement of the GAs and their subsequently produced metabolites was mainly acropetal. They accumulated in greatest quantity in the apical extremities of the shoot. Overall, the extent to which GA₂₀ was metabolished in le seedlings was considerably less than in Le pea seedlings. Although all le tissues contained significantly less [³H]GA₁ than their Le counterparts, phenotypic effects of the le mutation were apparent only on internode and tendril development. Increased tissue growth, consequent upon GA treatment, was also apparent only in the internodes and tendrils of le plants. For internodes, GA₁ content determined the mid-logarithmic-phase growth rate and, consequently, final length. For tendrils, GA₂₀ rather than GA₁ may be the primary stimulatory agent.Abbreviations GA gibberellin - HPLC high-performance liquid chromatography - 1–6 consecutive developmental numbering system for plant tissues/organs as shown in Fig. 1 The author gratefully acknowledges financial support from Imperial Chemical Industries, Plant Protection, Jealott's Hill, Bracknell, Berks., UK and the Science and Engineering Research Council.     

Ethylene metabolism in Pisum sativum L.

The possible role of C₂H₄ metabolism in mediating the responses of plants to C₂H₄ is re-examined. It is demonstrated that (i) the effects of inhibitors upon C₂H₄ action do not correspond with their effects on metabolism, (ii) elicitors of C₂H₄ effects do not have appropriate effects on C₂H₄ metabolism, (iii) inhibitors of C₂H₄ metabolism do not affect the response of plants to C₂H₄. It is concluded that metabolism of C₂H₄ is not linked to the mode of action of the growth regulator.Abbreviations DTC sodium diethyldithiocarbamate - FW fresh weight     

Synthesis of Phytochelatins and Homo-Phytochelatins in Pisum sativum L

In the roots of pea plants (Pisum sativum L.) cultivated with 20 [mu]M CdCl2 for 3 d, synthesis of phytochelatins [PCs or ([gamma]EC)nG, where [gamma]EC is [gamma]glutamylcysteine and G is glycine] and homophytochelatins [h-PCs, ([gamma]EC)n[beta]-alanine] is accompanied by a drastic decrease in glutathione (GSH) content, but an increase in homoglutathione (h-GSH) content. In contrast, the in vitro activity of GSH synthetase increases 5-fold, whereas h-GSH synthetase activity increases regardless of Cd exposure. The consititutive enzyme PC synthase, which catalyzes the transfer of the [gamma]-EC moiety of GSH to an acceptor GSH molecule thus producing ([gamma]EC)2G, is activated by heavy metals, with Cd and Cu being strong activators and Zn being a very poor activator. Using h-GSH or hm-GSH for substrate, the synthesis rate of([gamma]EC)2[beta]-alanine and [gamma]EC)2-serine is only 2.4 and 0.3%, respectively, of the sythesis rate of ([gamma]EC)2G with GSH as substrate. However, in the presence of a constant GSH level, increasing the concentration of h-GSH or hm-GSH results in increased synthesis of ([gamma]EC)2[beta]-alanine or ([gamma]EC)2-serine, respecively; simultaneously, the synthesis of ([gamma]EC)2G is inhibited. [gamma]EC is not a substrate of PC synthase. These results are best explained by assuming that PC synthase has a [gamma]EC donor binding site, which is very specific for GSH, and a [gamma]EC acceptor binding site, which is less specific and accepts several tripeptides, namely GSH, h-GSH, and hm-GSH.