Anther Culture of Lolium temulentum, Festuca pratensis and Lolium x Festuca Hybrids. II. Anther and Pollen Development In Vivo and In Vitro

The various pathways of pollen development were investigatedin cultured anthers of Lolium temulentum, Festuca pratensisand the L. multiflorum x F. pratensis hybrid ‘Elmet’.In all three, development from the vegetative cell was the predominantpathway of pollen callus development. However, there were characteristicdifferences in the behaviour of the generative cell. In L. temulentumit remained attached to the pollen wall and degenerated, whereasin F. pratensis it divided. In ‘Elmet’ it detachedfrom the pollen wall and remained undivided. Both polarizedand unpolarized partitioned calluses were observed. Developmentof the fusion product of the vegetative and generative nucleiwere also observed in anthers of L. temulentum. Anomalous grainswere not found to be major source of pollen calluses. Sections of anthers of L. temulentum were used to investigatethe origin of S pollen grains, the small pale-staining grainswhich denote pollen dimorphism. Such grains form out of contactwith the tapetum and are therefore determined before or duringmeiosis (i.e. before harvest of anthers for culture). Sectionswere also used to demonstrate the influence of the durationof pretreatment on the development of the middle layer of theanther wall. Festuca pratensis, Lolium temulentum, Lolium x Festuca, anther culture, haploid, microspore, pollen     

Callus Cultures from Zygotic Embryos of Costus speciosus and Their Morphogenetic Responses

Soft, nodular and hard types of calli were initiated on mature zygotic embryo explants of two tetraploid clones of Costus speciosus, of which, only the hard calli were amenable to morphogenetic responses. The two clones differed in their growth regulator requirements both for the initiation of calli and for shoot regeneration. De novo formation of both shoot bud meristems and somatic embryoids were observed. Latter were encased partially or fUlly by coleoptilar sheath. Embryoids could be isolated as discrete units. On maturity, a stock like appendage developed from the base and finally embryoids got detached from the subtending tissue. Both shoot-bud meristems and somatic embryoids developed into complete plantlets, the former upon sequential transfer of calli on Schenk and Hildebrandt’s (SH) basal medium containing lower levels of growth hormones, while the latter only on basal medium. These culture regenerants were subsequently transferred to the field. The morphogenetic behaviour of these two tetraploid clones reflects their marked genotypic difference inspite of their same ploidy status.     

大葱成熟胚离体培养植株再生

以大葱成熟胚为外植体,就激素浓度对其愈伤组织和不定芽的诱导情况进行了初步探讨。结果表明,2,4-D(1.0mg/L-2.0mg/L) 6-BA(0.5mg/L-2.0mg/L)为诱导愈伤组织的适宜组合范围,不定芽的诱导以6-BA（1.0mg/L-2.0mg/L) NAA)65mg/L效果最好,不定芽在MS0上可正常生根,发育成完整植株。     

In Vitro Selection for Salt Tolerance in Brassica juncea L. Using Cotyledon Explants, Callus and Cell Suspension Cultures

Salt-tolerant Brassica juncea L. cell lines or plants have beenselected by screening callus pieces, cell suspension culturesand cotyledon explants in vitro on high concentrations of NaCl.Callus-based selection was unsatisfactory, as only two out ofseven isolated clones retained tolerance after 3 months of subcultureon NaCl-free medium. Selections made via plated cell suspensionswere found to be more stable for salt-tolerance. AH selectedtolerant cell lines, however, failed to regenerate plantlets.A third selection method, employing cotyledon explants was basedon their high potential for regenerating multiple shoots. Outof a total of 2620 explants cultured on high salt media, threesurvived, showed sustained callus proliferation and each regeneratedone shoot. The salt-selected shoots withstood the stabilitytest after 3 months of growth and axillary bud multiplicationon NaCl-free medium. While one of these somaclones was morphologicallyabnormal and sterile, the other two could be reared to maturitywith normal seed set. Brassica juncea, tissue culture, in vitro selection, salt-tolerance, plant regeneration     

In Vitro Morphogenesis in Nardostachys jatamansi DC.: Shoot Regeneration from Callus Derived Roots

Callus cultures of Nardostachys jatamansi DC. maintained onMurashige and Skoog's medium containing 3.0 mg 1^–1 of-naphthaleneacetic acid and 0.25 mg 1^–1 of kinetin whenshifted to medium containing 0.25–1.0 mg 1^–1 ofindole-3-acetic acid or indole-3-butync acid showed profuserhizogenesis. The callus-regenerated roots when transferredto medium containing 2.0–6.0 mg 1^–1 of kinetin producedshoot buds. The de novo shoot bud regeneration took place eitherdirectly from cortical cells or from the inner stelar region.In addition, direct, concomitant root-shoot development wasalso observed. Nardostachys jatamansi, organogenesis, root-buds     

In Vitro Shoot Regeneration from Callus, Leaf Axils and Petioles of Sugar Beet (Beta vulgaris L.)

Procedures are described for producing axillary shoots fromseedling apices and adventitious shoots from petioles and leaf-derivedcallus of sugar beet cultivars. The rate of adventitious shootregeneration from petioles was influenced by temperature, BAPconcentration of the medium, and the time in culture of theseedling apices from which the petioles were excised. Petiolesectioning confirmed that adventitious shoots originated inthe sub-epidermal parenchyma. Two distinct types of callus wereproduced from leaf explants, but only white friable callus wascapable of shoot development. This callus developed from browntissue and was composed of thin-walled cells with dense cytoplasmand prominent nuclei. Green compact callus with thick-walledlignified cells developed from green tissue, but did not produceshoots. Successful seed sterilization and shoot regenerationfrom petiole explants and callus was cultivar-dependent. Adventitiousshoots were rooted and successfully transplanted to pottingcompost under glasshouse conditions. Key words: Adventitious shoots, axillary shoots, callus, sugar beet (Beta vulgaris L.)     

Starch Metabolism, Respiration, and Shoot Formation in Tobacco Callus Cultures

The starch content of shoot-forming and non-shoot-forming tobacco callus cultured in light and darkness was determined. A variety of carbohydrates and cytokinins incorporated into the culture medium were effective in bringing about starch accumulation and shoot formation in the tissue. In addition, the respiratory activity of the callus, grown in the presence or absence of gibberellic acid, was measured. A strong correlation between the starch content of the tissue, its rate of respiration, and shoot formation was observed.     

Reproductive Development in Lolium temulentum L.: Spike Morphogenesis and Grain Set Limitations

Reproductive development in cereals is not easy to investigatebecause their quantitative response to environmental factorsmakes it difficult to synchronize the plants. In this paper,one of our aims was to assess whether Lolium temulentum strainCeres, a qualitative long-day grass, could serve as a modelof reproductive development for cereals. The morphological patternsfrom floral transition to seed set were studied. A flowering scale was established to evaluate developmentalrate during spike morphogenesis. Apex growth was found to increaseaccording to biphasic kinetics; double ridge appearance markedthe beginning of an exponential phase. Developmental progression and apical growth rate were both increasedby giving repeated long days. In contrast, the final numberof lateral spikelets (20–25) could not be manipulatedafter the beginning of long-day treatment. When plants were kept in continuous light from the beginningof induction, double ridges appeared on the fifth 24 h cycle.Spikelet initiation began in the upper mid-part of the spike,and then extended acro- and basipetally. The phase of spikeletinitiation lasted 6 d, with l?5 to 1?9 spikelets being produceddaily. Within each spikelet, florets were initiated at an averagerate of l?3 primordia per day and developed acropetally. Thefirst signs of apical site degeneration were observed in themost developed upper spikelets just before heading. Ear emergenceoccurred between the 20th and 25th cycles of continuous light;anthesis was observed 6 or 7 d later. The proportion of floretssetting grain averaged about 40%. Grains were produced mainlyin the lower spikelets while the upper mid-part of the inflorescenceshowed a much lower fertility rate. Complex developmental gradients described in this paper suggestthat L. temulentum could serve as a model of reproductive developmentin cereals, with the added advantage of flowering in responseto a single long-day. Key words: Lolium temulentum L., spike morphogenesis, spikelet number, floret number, grain set     

In Vitro Culture of Subanguina picridis in Acroptilon repens Callus,Excised Roots,and Shoot Tissues

The knapweed nematode Subanguina picridis is a foliar parasite that is of interest as a biological weed control agent of Russian knapweed. Attempts were made to culture the nematode in callus, excised roots and in shoots derived from roots of Russian knapweed. In callus tissues, the nematode developed from second-stage juvenile to adult but failed to reproduce; it developed only to the fourth stage in excised roots. However, S. picridis was successfully cultured in vitro in shoots derived from roots. The nematode induced galls on the leaves, petioles, and shoot apices and developed and reproduced inside the galls. Gibberellic acid increased the development rate of the nematode and promoted the formation of males. This is the first gnotobiotic culture of a nematode used for biological weed control.     

The typification of Lolium perenne L. and Loliutn temulentum L. (Poaceae)

LOOS, B. P. & JARVIS, C. E., 1992. The typification of Lolium perenne and Lolium temulentum (Poaceae). The typification of the Linnaean species Lolium perenne and Lolium temulentum has been studied. Lolium perenne is typified by material in LINN, as proposed by Terrell, but it has been necessary to select a lectotype for L. temulentum , and material in the Burser herbarium (UPS) has been chosen for this purpose. The study shows that although Linnaeus used awns as a diagnostic character to distinguish the two species, he was aware of the intraspecific variability in this character.     

The distribution of acid invertase in developing leaves of Lolium temulentum L.

The levels of invertase (E.C. 3.2.1.26) activity were measured throughout the development of the fourth leaf of Lolium temulentum. No neutral invertase activity was detected. Soluble acid invertase activity fell during leaf extension but rose again after ligule formation. This rise continued into senescence and was accompanied by the appearance of activity in the insoluble fraction. Evidence is presented that the insoluble activity was not an artefact of preparation, and that it represented an extracellular acid invertase. Fractionation of soluble invertase by gel filtration showed the appearance of a high molecular weight form at the time when insoluble activity was rising. The relationships between the different forms of the enzyme are discussed, together with their roles in leaf development.