Shoot development ofAucuba japonica I. Morphological study

The shoot development ofAucuba japonica was studied morphologically. The shoot shows dichasial branching in connection with the formation of a terminal inflorescence and shows a decussate phyllotaxis even in the reproductive phase. The sequence of initiation of successive foliar appendages is very precise, hence the foliage leaf, scale leaf and bract can be compared with each other even at their stages of initiation. In the stage of proximal foliage leaf formation the shoot apex is flat, while in the stage of formation of distal foliage leaves, bud scales and proximal bracts, it becomes concave. In the stage of formation of distal bracts the apex becomes domed. Plastochron durations are relatively long in the vegetative phase in comparison with other plants, and the duration from initiation of the first pair of appendages to that of the second is about one and a half months. Both male and female inflorescences exhibit basically a thyrsoid type of monotelic synflorescence.     

Shoot production per responsive leaf explant increases exponentially with explant organogenic potential in Nicotiana species

As part of the effort to develop optimal plant varieties for the production and molecular farming of plant-made pharmaceuticals, this study evaluated shoot organogenic potential of a total of 115 Nicotiana accessions, representing 53 species. To induce shoots, leaves from seedling grown in vitro were cut into pieces, cultured on shoot-induction medium under low light for 3 weeks, and then subcultured onto the same medium for another 4 weeks under normal light. Statistical analysis detected significant differences among the 115 accessions for the percentage of leaf explants producing shoots and the number of shoots produced per responsive leaf explant. Importantly, regression analysis also found an exponential relationship between the number of shoots produced per responsive leaf explant and the percentage of leaf explants producing shoots. The number of shoots produced per responsive leaf explant increased rather slowly, ranging from zero to around five, as the percentage of leaf explants producing shoots increased from 0 to 80%, but the increase became dramatic as the percentage increased from 80% to 100%, reaching as high as 35 shoots per responsive leaf explant. This exponential relationship is the first of its kind to be established in plant regeneration studies using either organogenesis or somatic embryogenesis systems. A possible mechanism that governs the establishment of the exponential relationship is discussed.Abbreviations 2ip 6-(,-Dimethylallylamino)-purine - BA Benzylaminopurine - IAA Indole-3-acetic acid - LS Linsmaier and Skoog - MS Murashige and Skoog - PI Plant introduction number - PMP Plant-made pharmaceuticals - SIM Shoot induction medium - USDA US Department of Agriculture     

Shoot development and production studies of phragmites Australis (Cav.) Trin. Ex Steudel in Scottish Lochs

Studies were made on the development and production of Phragmites australis (Cav.) Trin. ex Steudel (= Phragmites communis Trin.) stands from Forfar Loch-polytrophic, Balgavies Locheutrophic and Loch of the Lowes-mesotrophic. Differences were detected in the shoot height, shoot dry weight, standing crop biomass, leaf area index, crop growth rate and net efficiency of solar energy conversion among the three lochs. Such differences were partly due to the different nutrient levels of the three locations. The percentage ratio of leaf to shoot weight, leaf area ratio, relative growth rate and net assimilation rate for the reed stands in the three lochs were also calculated and were compared with values from other study areas.     

Transformation and regeneration of loblolly pine: shoot apex inoculation with Agrobacterium

Loblolly pine (Pinus taeda L.) is the mostimportant tree species in US commerce and has much to gain through geneticengineering. This species can be transformed using particle bombardment andAgrobacterium; however, the regeneration of plants fromtransgenic tissues has been difficult and the recovery of transgenic plants hasbeen rare. A shoot-based and genotype-independent transformation methodemploying Agrobacterium tumefaciens was used to facilitaterecovery of plants and permit the transformation of elite germplasm. Shootsfrom4–6 week old seedlings and adventitious shoots from culture wereinoculated with A. tumefaciens EHA101 (pGUS3), or EHA105(pSSLa.3), subjected to selection and regenerated. Shoots that survivedexhibited expression of the uidA gene (GUS) in a patterncharacteristic of the either the CaMV35S promoter (pGUS3), or the larch RbcSpromoter (pSSLa.3) transferred. Recovered plants were screened using PCRamplification. Southern DNA analyses and amplification of the T-DNA borderjunction confirmed genomic integration of both transferreduidA and nptII genes. In this proofofconcept study, the overall recovery of P. taeda shoots wasfair (10–20%), while recovery of intact rooted plants was poor (>1%)due to difficulty in rooting. Recovery of intact rooted plants from inoculatedshoots of P. eldarica and P. radiatawas more efficient (10–30%). The addition of a shoot multiplication stepand effective rooting protocols will improve the efficiency of this genotypeindependent transformation method in P. taeda, and inotherPinus spp.     

Effects of red and far-red light on cell division in the shoot apex ofSilene coeli-rosa L.

Summary 28-day-old plant ofSilene coeli-rosa were exposed at 1,700 hours to 5 or 10 minutes red light, 5 or 10 minutes far-red light, red followed by far-red, far-red followed by red or maintained in darkness. Measurements of the proportions of cells with the 2 C and 4 C amounts of DNA in the shoot apex of the plants, sampled at 2,000 hours, showed that far-red light promoted an increase in the G2 proportion whereas red light resulted in an increase in the G1 proportion of the cell cycle, relative to the dark controls. Moreover these changes were red, far-red reversible. All light treatments resulted in increases in the mitotic index in the apex compared with the dark controls, suggesting increases in the growth rate. The data implicate phytochrome in a low energy response and suggest that, in the shoot apex, G1 is shortened markedly following exposure to farred light, whilst G2 is shortened the most following exposure to red light. The results are discussed in relation to flower-initiation.     

Micropropagation of black pepper (Piper nigrum Linn.) through shoot tip cultures

Summary The morphogenetic potential of shoot tip explants of black pepper (Piper nigrum) was investigated and an effective multiple-shoot propagation method is described. Various combinations of media, growth regulators and sterilization treatments were compared. Problems with establishment in tissue culture sometimes occurred, probably caused by endogenous pathogens associated with tissue exudates. The best establishment and proliferation of shoot tip explants was obtained on MS medium containing 1.5 mg l^–1 BAP alone; subsequent growth and development of lateral branches was best on media containing 1.5 mg l^–1 BAP plus 3.0 mg l^–1 IBA. Adenine sulphate inhibited the number of explants showing regeneration but increased the number of shoot buds per regenerating explant. Shoots were rooted on a 50% strength medium containing 1mg l^–1 NAA.Abbreviations AdSO₄ adenine hemisulphate - BAP 6-benzylaminopurine - IBA indole-3-butyric acid - NAA napthaleneacetic acid     

Morphological and phenological characteristics of leaf development ofDurio zibethinus Murray (Bombacaceae)

The morphological and phenological characteristics of leaf development ofDurio zibethinus Murray were investigated at an experimental field of Universiti Pertanian Malaysia (UPM) in Selangor. Proportionality was observed in the relations of leaf length to leaf width and of leaf area to the product of leaf width and length. The proportionality was explained from the similarity of leaf shape. New leaves emerged continuously, but the number of new leaves fluctuated seasonally. The emergence of leaves was inhibited by the flower bud formation. In the survival curves of leaves, the relative fall rate was lower at the early stage of leaf development than at the late stage. Leaf longevity of 100 to 133 days was low and leaf expansion period of two weeks was short in comparison with the published data on tropical trees. From the ecophysiological viewpoint, the leaf survival strategy of the present species was discussed: the present species manages to set up a photosynthetic system in a short period by the rapid leaf growth; the lower leaf longevity is advantageous to reaching more frequently high photosynthetic production by newly emerged leaves.     

Inhibition of growth and synchronised cell division in the shoot apex in relation to flowering in Silene

When plants of Silene coeli-rosa (L.) Godron were induced by seven long days, then exposed to darkness for 48 h before being returned to short days, they went on to initiate flowers with a delay of about 2 d. The synchronisation of cell division which normally occurs before flower initiation was suppressed, showing that it is not essential for flowering. Periods of darkness of up to 240 h inhibited apical growth and leaf initiation but did not prevent eventual flowering in short days. The commitment of the apex to flower was therefore maintained while apical growth was inhibited.Abbreviations SD short day(s) - LD long day(s)     

Mean rate of DNA replication and replicon size in the shoot apex ofSilene coelirosa L. During the initial 120 minutes of the first day of floral induction

Summary 28-day-old plants ofSilene coeli-rosa were exposed, at 1,700 hours, to long day (LD) conditions comprising light of low fluence rate provided by tungsten bulbs, or maintained in darkness as short day (SD) controls. All plants were exposed at 1,700 hours to tritiated-(methyl-³H)-thymidine for 30, 45, 60, 90, or 120 minutes. Apical domes were isolated and prepared as fiber autoradiographs from which replicon size and rates of DNA replication, per single replication fork were recorded. In SD, replicon size was between 15–20 m and exposure to LD conditions altered neither replicon size nor the pattern of deployment of replicons during S-phase relative to the SD controls. However, the mean rate of replication in LD was 8.7 m h^–1 compared with 5.2 m h^–1 in SD. Thus, exposure to LD resulted in a 1.7-fold increase in the rate of DNA replication relative to the SD controls. This rapid increase in replication rate, detectable within 30 minutes of the start of the LD is discussed in relation to changes known to occur to the cell cycle inSilene during the first day of floral induction.     

Identification of genes expressed in the shoot apex ofBrassica campestris during floral transition

The vegetative-to-floral transition ofBrassica campestris cv. Osome was induced by vernalization. Poly(A)+RNA was isolated from the transition shoot apex after 6 weeks of vernalization, the floral apex after 12 weeks of vernalization and the expanded leaves just before vernalization, and cDNAs were synthesized. These cDNAs were used for subtraction and differential screening to select cDNA preferentially present in the transition and floral apices. Nucleotide sequences of the resulting 14 cDNA clones were determined, and northern blot analysis was carried out on six cDNAs. Two cDNA clones which did not show significant similarity to known genes were shown to be preferentially expressed in the floral apex.     

Improved guggulsterone production from sugars, precursors, and morphactin in cell cultures of Commiphora wightii grown in shake flasks and a bioreactor

Cell cultures of Commiphora wightii (Arnott.) Bhandari were grown in shake flasks and a bioreactor and an increase in guggulsterone accumulation up to 18 μg l⁻¹ was recorded in cells grown in the production medium containing a combination of sucrose:glucose (4% total), precursors (phenylalanine, pyruvic acid, xylose, and sodium acetate), morphactin, and 2iP. A yield of 10 g l⁻¹ biomass and ∼200 μg l⁻¹ guggulsterone was recorded in a 3-l flask and in a 2-l stirred tank bioreactor compared with 6.6 g biomass and 67 μg l⁻¹ guggulsterone in 250-ml flasks. Increased vessel size was correlated with increased biomass and guggulsterone accumulation. 2iP alone was not effective for biomass and guggulsterone accumulation in cell cultures of C. wightii.     

<Emphasis Type="Italic">EcFLO</Emphasis>, a<Emphasis Type="Italic"> FLORICAULA</Emphasis>-like gene from<Emphasis Type="Italic"> Eschscholzia californica</Emphasis> is expressed during organogenesis at the vegetative shoot apex

FLORICAULA/ LEAFY-like genes were initially characterized as flower meristem identity genes. In a range of angiosperms, expression occurs also in vegetative shoot apices and developing leaves, and in some species with dissected leaves expression is perpetuated during organogenesis at the leaf marginal blastozone. The evolution of these expression patterns and associated functions is not well understood. We have isolated and characterized a FLORICAULA-like gene from California Poppy, Eschscholzia californica Cham. (Papaveraceae), a species belonging to the basal eudicot clade Ranunculales. EcFLO encodes a putative 416-amino-acid protein with highest similarity to homologous genes from Trochodendron and Platanus. We show that EcFLO mRNA is expressed during the vegetative phase of the shoot apical meristem and in developing dissected leaves in a characteristic manner. This pattern is compared to that of other eudicots and discussed in terms of evolution of FLORICAULA expression and function.     

Impact of epidermal leaf mining by the aspen leaf miner (Phyllocnistis populiella) on the growth, physiology, and leaf longevity of quaking aspen

The aspen leaf miner, Phyllocnistis populiella, feeds on the contents of epidermal cells on both top (adaxial) and bottom (abaxial) surfaces of quaking aspen leaves, leaving the photosynthetic tissue of the mesophyll intact. This type of feeding is taxonomically restricted to a small subset of leaf mining insects but can cause widespread plant damage during outbreaks. We studied the effect of epidermal mining on aspen growth and physiology during an outbreak of P. populiella in the boreal forest of interior Alaska. Experimental reduction of leaf miner density across two sites and 3 years significantly increased annual aspen growth rates relative to naturally mined controls. Leaf mining damage was negatively related to leaf longevity. Leaves with heavy mining damage abscised 4 weeks earlier, on average, than leaves with minimal mining damage. Mining damage to the top and bottom surfaces of leaves had different effects on physiology. Mining on the top surface of the leaf had no significant effect on photosynthesis or conductance and was unrelated to leaf stable C isotope ratio (delta(13)C). Mining damage to the bottom leaf surface, where stomata are located, had significant negative effects on net photosynthesis and water vapor conductance. Percent bottom mining was positively related to leaf delta(13)C. Taken together, the data suggest that the primary mechanism for the reduction of photosynthesis by epidermal leaf mining by P. populiella is the failure of stomata to open normally on bottom-mined leaves.     

Gradual Depletion of 2,4-D in the Culture Medium for Indirect Shoot Regeneration from Leaf Explants of Camellia sinensis (L.) O. Kuntze

Adventitious shoot regeneration via callus phase from in vitro leaf explants is reported for the first time in tea. Callus was obtained on Murashige and Skoog medium supplemented with varied concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) (2.5, 5.0, 7.5 and 10.0 mg/l). Rhizogenesis was observed at all concentrations of 2,4-D. Adventitious shoot buds developed indirectly on leaf explants after prolonged culture for 16 weeks on medium supplemented with 10.0 mg/l 2,4-D. GC analysis of the medium and the tissues at different stages of development showed that specific levels of 2,4-D in the tissue were responsible for morphogenesis. Shoot buds developed on rhizogenic calli, only when 2,4-D declined to undetectable or negligible concentrations in the tissue probably due to detoxification and metabolism. Alternatively, shoot buds could also be evoked when rhizogenic calli were transferred to medium supplemented with low concentration of 2,4-D (1.5 mg/l). The adventitious nature of the shoots was confirmed through histological studies.     

Arrangement of cortical microtubules in the shoot apex of Vinca major L.

The arrangements of cortical microtubules (MTs) and of cellulose microfibrils in the median longitudinal cryosections of the vegetative shoot apex of Vinca major L., were examined by immunofluorescence microscopy and polarizing microscopy, respectively. The arrangement of MTs was different in the various regions of the apex: the MTs tended to be arranged anticlinally in tunica cells, randomly in corpus cells, and transversely in cells of the rib meristem. However, in the inner layers of the tunica in the flank region of the apex, cells with periclinal, oblique or random arrangements of MTs were also observed. In leaf primordia, MTs were arranged anticlinally in cells of the superficial layers and almost randomly in the inner cells. Polarizing microscopy of cell walls showed that the arrangement of cellulose microfibrils was anticlinal in tunica cells, random in corpus cells, and transverse in cells of the rib meristem; thus, the patterns of arrangement of microfibrils were the same as those of MTs in the respective regions. These results indicate that the different patterns of arrangement of MTs and microfibrils result in specific patterns of expansion in the three regions. These differences may be necessary to maintain the organization of the tissues in the shoot apex.Abbreviations MT(s) microtubule(s) - lp length of the youngest leaf primordium     

Base temperature and heat units for leaf flushing emission and growth ofHevea brasiliensis Muell. Arg.

In this study we determined the base temperature and Heat units for leaf flushing initiation and growth of RRIM-600 and GT1Hevea clones. The minimum base temperature was found to be approximately 16° C for leaf flushing emission and 19° C for shoot growth in terms of height. Approximately 420 degree days at 16° C as base temperature is required for successive leaf flushing initiation. Linear equations with correlation coefficients above 0.95 allow an estimation of the height increase from the accumulated degree days, corrected or otherwise for photoperiod. The highest correlation coefficients demonstrated a positive effect of the photoperiod among the factors influencing the shoot growth.     

Hyphal tip ultrastructure ofAspergillus nidulans andAspergillus giganteus and possible implications of woronin bodies close to the hyphal apex of the latter species

Summary The hyphal tip ultrastructure ofAspergillus nidulans andAspergillus giganteus indicates that their apical organization is very similar to that found in other filamentous fungi. Both species have an area immediately behind the hyphal apex free of all large organelles and containing a high concentration of vesicles. InA. giganteus only one size class of vesicle is clearly evident, with a mean diameter of 72 nm. InA. nidulans two size classes of vesicle were found, with mean diameters of 75 nm and 31 nm. A Spitzenkörper is evident inA. nidulans as an area very close to the tip containing only the smaller vesicles. InA. giganteus one or more apparently mature Woronin bodies were found within the first 1 m of some hyphal apices. The possible significance of their presence is discussed.     

Microspectrophotometric measurements of DNA by automated computerized scanning in cycling cells of theChrysanthemum segetum shoot apex

Summary A new technique of exploitation of the data was proposed after DNA scanning microdensitometry. By using all of the measurements obtained from the seriated sections of a single nucleus, this method made it possible to estimate six characteristic parameters during the different phases of the cell cycle in the various shoot apical cells. The cells whose rate of proliferation was the highest showed the biggest variations of their nuclear and nucleolar volumes during the cell cycle. In the axial zone, where the cells have a slow cell cycle and display the longest duration of the G₁ phase, the volume occupied by dispersed DNA was greater than in the cells of the lateral zone and of the rib meristem, where the cell cycle and the G₁ phase were short. No matter what the cell type, the proportion of the dispersed and condensed DNA varied little when the G₁ and G₂ phases were compared. In the Z phase, characterized by a decondensation of the DNA, the mean DNA amount was 3.4 C. The evolution of the nuclear density during the interphase was also estimated. It is demonstrated that the main feature of the shoot apex zonation was the decondensation of the condensed DNA in the axial zone in both the G₁ and G₂ phases.     

Ageing of theSilene coeli-rosa L. shoot apex under non-inductive conditions: Changes in morphology,mitotic index,and polypeptide composition

Summary Ageing was studied in the shoot apex of the long day plant,Silene coeli-rosa by maintaining it in non-inductive short day conditions for 170 days. The dimensions, the zonation, and the polypeptidic pattern of the shoot apex, and the rate of leaf initiation were altered in 170-day-old plants compared with young plants grown under the same conditions (28 days). In aged plants, the number of cells increased in all shoot apical zones and, notably, the mitotic index increased in the axial zone; however, the rate of leaf initiation slowed down. These changes showed some similarities to those during the intermediate phase in quantitative photoperiodic species. The two-dimensional mini-gel electrophoretic study of protein extracts from shoot apices of young and ageing plants maintained in non-inductive conditions revealed 489 common polypeptidic spots, 13 unique to the young state and 24 new ones specific to aged plants. The spots characteristic for each state represented only 3.6% of the total identified polypeptides, but apical development under non-inductive conditions was characterized by qualitative changes in the polypeptide complement.Abbreviations C nuclear DNA complement - 1D gel first dimension gel - LD long day - NEPHGE non equilibrium pH gradient electrophoresis - SD short day - SDS sodium dodecyl sulfate - TCA trichloracetic acid