Hollow heart of pea (Pisum sativum)

Book reviewed in this article: The Way Ahead in Plant Breeding. Proceedings of the Sixth Congress of Eucarpia. Edited by F. G. H. Lupton , G. Jenkins and R. Johnson . Basic Electron Microscope Techniques. By M. A. Hayat . The Logit Transformation (with special reference to its uses in bioassay). By W. D. Ashton . Families of Frequency Distributions. By J. K. Ord .     

Verticillium wilt of pea (Pisum sativum)

A wilt disease of garden pea (Pisum sativum) caused by Verticillium dahliae is described and the range of pathogenicity of the isolate investigated. It is pathogenic to potato, sweet pea, antirrhinum and broad bean and isolates of V. dahliae from potato, lucerne and sweet pea and V. albo-atrum from lucerne are pathogenic to pea. Since the most common disease symptoms, acropetal progression of chlorosis and necrosis of the leaves followed by premature defoliation are indistinguishable from natural senescence, it is probable that disease and senescence symptoms are confused in the field. The premature defoliation results in marked reduction in green leaf area, leaf dry weight and pod yield.     

Embryogenesis of the pea (Pisum sativum)

Summary The most striking internal feature of the suspensor cells inPisum is the abundant occurrence of a plastid containing spherical bodies consisting of intertwined bundles of tubules. These tubular complexes are not typical prolamellar bodies and they are not converted into grana. Cytochemical reactions indicate that they are proteinaneous. The participation of this plastid in the possible nutritional function of the suspensor is discussed but it is pointed out that critical experimental evidence is needed before the role of the suspensor and its contents in embryogenesis can be understood.Supported by a grant from the Australian Research Grants Committee.     

Pisatin metabolism in pea (Pisum sativum L.) cell suspension cultures

Cell suspension cultures were established from germinating pea (Pisum sativum L.) seeds. This cell culture, which accumulated pisatin, consisted mostly of single cells containing a few cell aggregates. The cells responded to treatment with a yeast glucan preparation with transient accumulation of pisatin in both cells and culture media. Addition of pisatin to cell cultures resulted in increased synthesis of pisatin. Phenylalanine ammonia-lyase, chalcone synthase and isoflavone reductase activities were present in untreated cells. Upon treatment with an elicitor preparation the activities of the first two enzymes showed a rapid, transient increase up to 20 hours after treatment. Isoflavone reductase showed a major and minor peak at 16 and 36 h, respectively, after elicitor treatment. The time course of the enzyme activity and pisatin accumulation is consistent with an elicitor-mediated response.Abbreviations CHS chalcone synthase - 2,4-D 2,4-dichlorophenoxyacetic acid - IBA indole-3-butyric acid - IFR isoflavone reductase - 2iP 6-(dimethylallylamino)-purine - MS Murashige & Skoog basal salt medium - PAL phenylalanine ammonia-lyase - PMSF phenylmethylsulfonyl fluoride - POPOP 1,4-bis-2-(4-methyl-5-phenyloxazolyl)-benzene - PPO 2,5-diphenyloxazole     

Boron alleviates aluminum toxicity in pea (Pisum sativum)

One important target of boron (B) deficiency and aluminum (Al) toxicity is cell wall. Thus we studied the hypothesis that B is capable of alleviating Al toxicity in pea (Pisum sativum). Short-term and prolonged Al exposure to pea roots at different B levels was carried out on uniform seedlings pre-cultured at a low B level. When seedlings with a low B level were supplied with or without B for 1 and 2 days before 24 h Al exposure, roots were longer while root diameter was thinner after B addition especially for 2 days even with exposure to Al; root elongation was inhibited while root diameter was enlarged by Al exposure. Callose induction by Al toxicity was higher with B added, but this was reversed after the removal of the cotyledons. Hematoxylin staining was lighter in the root tips given B, and Al content in the root tips and cell walls dropped after exposure to B. This indicates that B alleviated Al toxicity in the root tips during short-term Al exposure by decreasing Al binding in root cell walls. An increase in chlorophyll and biomass and reduced chlorosis were found at the higher level of B during prolonged Al treatment, which was coincided with the decreased Al contents, indicating that B alleviated Al toxicity to shoots. B supplementation alleviates some of the consequences of Al toxicity by limiting some Al binding in cell walls, resulting in less injury to the roots as well as less injury to the shoots.     

High frequency adventitious shoot regeneration from immature cotyledons of pea (Pisum sativum L.)

A procedure has been developed which allows high frequency adventitious shoot regeneration from immature cotyledons of pea. Prolific shoot regeneration occurred following an initial callus growth on a Murashige and Skoog (MS) medium containing 0.5 mg/l 6-benzylaminopurine (BAP) and 4 mg/l -naphthaleneacetic acid (NAA). Cotyledon expiants proximal to the embryonic axis had the highest regeneration potential, however, the presence of an embryonic axis inhibited adventitious shoot regeneration. Addition of silver nitrate (AgNO₃) to the medium did not promote the number of regenerated shoots but resulted in shoots with well developed tendrils and large stipules which had a reduced rooting capacity. Regenerated shoots rooted readily (80–90%) in half strength MS medium containing 1 mg/l indole-butyric acid (IBA) and further established well in compost.     

Rapid multiple shoot production from cotyledonary node explants of pea (Pisum sativum L.)

Summary Multiple shoots were produced, from cotyledonary node explants of pea (Pisum sativum L.) cultured on MS medium containing 1 mg 1⁻¹ BAP. The number of buds formed could be increased by scraping the nodes before culture or by increasing the cytokinin concentation. However, cytokinin levels over 5 mg 1⁻¹ increasingly produced shoots that were vitrified and dificult to root. With all the genotypes tested, developing buds were visibile as soon as 5 days after culture and elongated shoots could be removed after 21 days., Histological studies indicated, that the buds and shoots were formed from superficial layers of tissue. The efficiency of this regeneration system compared favorably with previosly published methods. The rapid, genotype-independent, high frequency system described here may be of use in the production of transgenic pea plants.     

Localization of acid phosphatase activity in the apoplast of pea (Pisum sativum L.) root nodules grown under phosphorus deficiency

ACPase activity was localized in the apoplast of pea root nodules under phosphorus deficiency. Pea plants (Pisum sativum L. cv. Sze ciotygodniowy) where inoculated with Rhizobium leguminosarum bv. viciae 248 and were cultured on nitrogen-free medium with phosphate (−N/+P) or phosphate-deficient (−N/−P) one. In comparison with control nodules, P-deficient nodules showed the increase of ACPase activity in plant cell walls and the infection threads. The increase in bacterial ACPase activity under P-deficiency may reflect higher demand for inorganic phosphorus that is necessary for bacteria multiplication within the infection threads. The increase of ACPase activity in nodule apoplast under P stress may enlarge the availability of phosphate for plant and bacteria.     

Regeneration of shoots from pea (Pisum sativum) hypocotyl explants

A sequential indole-3-acetic acid (IAA)-zeatin treatment was applied to Pisum sativum hypocotyl explants, resulting in shoot formation from 50% of the explants. Shoots were easily rooted and transplantable plants could be obtained in 3 months. The method has been applicable to the 5 cultivars tested. Histological examination of explants suggests the shoots to be of de novo origin, which would make the system suitable for transformation experiments.     

Protochlorophyllide forms in non-greening epicotyls of dark-grown pea (Pisum sativum)

Low-temperature fluorescence emission spectra of 6.5-day-old dark-grown epicotyls of pea ( Pisum sativum ) revealed the presence of protochlorophyll(ide). The upper part of the epicotyl contained 30% of the protochlorophyll(ide) content per fresh weight found in pea leaves, whereas the lower part contained 3%. Three discrete spectral forms of protochlorophyll(ide) were clearly distinguished after Gaussian deconvolution of fluorescence excitation and emission spectra. Adding the satellite bands of the Qy_(0-0) transitions (the emission vibrational (Emv) bands with correlated amplitudes, gave the following delineation: Ex439–Em629–Emv684, Ex447–Em636–Emv700 and Ex456–Em650–Emv728. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) followed by immunodetection of whole tissue extracts of the epicotyl indicated the presence of NADPH-protochlorophyllide oxidoreductase (EC 1.3.1.33). Electron micrographs showed prolamellar bodies in at most 11 % of the plastid profiles of the epicotyl cells. These prolamellar bodies were smaller, and many of them showed less regular structure than those of the leaves. Taken together, the results indicate that the protochlorophyll(ide) in epicotyls is arranged in a different way than in leaves.     

Physiology of acclimation to salinity stress in pea (Pisum sativum)

Pea (Pisum sativum L.) seedlings were grown in half strength Hoagland solution and exposed to 0, 10, 25 mM NaCl and 2.5% PEG 6000 for 1 week (pre-treatment). Thereafter plants were exposed to 0 and 80 mM NaCl for 2 weeks (main treatment). The control plants were maintained in half strength Hoagland solution without NaCl. Various physiological parameters were recorded from control, pretreated and non-pretreated plants. There was no negative effect of the pre-treatments on growth (total fresh and dry matter production), and plants pre-treated with 10 mM NaCl had biomass accumulation equal to control plants. The beneficial effect of salt acclimation was also evident in the prevention of K⁺ leakage and Na⁺ accumulation, primary in roots, suggesting that here the physiological processes play the major role. 2.5% PEG 6000 was not as efficient as salt in enhancing salt tolerance and acclimation appears to be more related to ion-specific rather than osmotic component of stress. We also recorded an increase of the xylem K/Na in the salt acclimated plants. Therefore, the present study reveals that short-term exposure of the glycophyte P. sativum species activates a set of physiological adjustments enabling the plants to withstand severe saline conditions, and while acclimation takes place primary in the root tissues, control of xylem ion loading and efficient Na⁺ sequestration in mesophyll cells are also important components of this process.     

Danish Rhizobium leguminosarum strains nodulating 'Afghanistan' pea (Pisum sativum)

A wild pea ( Pisum sativum L.) native to Afghanistan normally known to be resistant to nodulation with European strains of Rhizobium leguminosarum was nodulated early and effectively in field soil in Denmark. Isolates from nodules formed effective nodules abundantly on 'Afghanistan' on reinfection under aseptic conditions. Five types differing in isoenzyme composition pattern were found among 15 isolates from 'Afghanistan' nodules. None were identical with the 'Tom' strain from Turkey, which also forms effective nodules with 'Afghanistan'. The five types were also different with respect to isoenzyme pattern from Rhizobium leguminosarum strains isolated from a modern pea variety cultivated in the same field.     

Transient protein expression in three Pisum sativum (green pea) varieties

The expression of proteins in plants both transiently and via permanently transformed lines has been demonstrated by a number of groups. Transient plant expression systems, due to high expression levels and speed of production, show greater promise for the manufacturing of biopharmaceuticals when compared to permanent transformants. Expression vectors based on a tobacco mosaic virus (TMV) are the most commonly utilized and the primary plant used, Nicotiana benthamiana, has demonstrated the ability to express a wide range of proteins at levels amenable to purification. N. benthamiana has two limitations for its use; one is its relatively slow growth, and the other is its low biomass. To address these limitations we screened a number of legumes for transient protein expression. Using the alfalfa mosaic virus (AMV) and the cucumber mosaic virus (CMV) vectors, delivered via Agrobacterium, we were able to identify three Pisum sativum varieties that demonstrated protein expression transiently. Expression levels of 420 ( 26.24 mg GFP/kgFW in the green pea variety speckled pea were achieved. We were also able to express three therapeutic proteins indicating promise for this system in the production of biopharmaceuticals.