Factors affecting Ctenarytaina thysanura oviposition on Boronia megastigma terminal shoots

Field observations and experiments showed that the attractiveness of Boronia megastigma terminal shoots for Ctenarytaina thysanura (Hemiptera: Psyllidae) oviposition was effected by the quality and quantity of terminal shoots, relative hardness of the terminal shoot combined with a closed or open architecture of the shoot, nitrogenous fertilizer applications and intraspecific competition between psyllids. Heavy feeding either by nymphal or adult psyllids greatly reduces the attractiveness of terminal shoots for oviposition. This deterrence was associated with the production of honeydew resulting in the development of sooty moulds. When all nymphs had developed to adults, a further 21 days was required for the shoots to be restored to their original condition and suitability for oviposition.     

Development and application of an integrated pest management programme for the psyllid,Ctenarytaina thysanura onBoronia megastigma in Tasmania

An integrated pest management programme for control of the psyllid,Ctenarytaina thysanura (Ferris & Klyver) (Hemiptera: Psyllidae) in commercial boronia plantations was developed and implemented from 1986–1989. The programme involved spraying boronia stems only with the systemic organophosphate mevinphos at peak incidence of late stage nymphs. At this time, the majority of parasitoids were pupating within mummified hosts. Consequently, actively feeding nymphs were potentially susceptible to mevinphos but the parasitoids were not. Stem application negated any contact effect on parasitoids and predators. The conservation of natural enemies subsequently reduced psyllid numbers and no insecticide has been required since 1989. In economic terms the benefit to cost ratio of the IPM programme was $22.40: $1.0 and this was 40 times greater than the conventional 10 demeton-S-methyl sprays used by growers prior to the study.     

Rapid Micropropagation of Five Cultivars of Mulberry

Multiple shoots were initiated from nodal and shoot tip explants collected from mature trees of Morus alba L. cultivars Chinese White, Kokuso-27 and Ichinose, and M. multicaulis Perr. cultivars Goshoerami and Rokokuyaso after 2 weeks of culture. Nodal explants were more responsive than shoot tip explants. Murashige and Skoog basal medium was found to be most suitable medium and 6-benzylaminopurine was the most effective cytokinin for shoot induction. Explants collected between April and September evoked better response than the explants collected between October and March. Shoots were multiplied by transferring nodal explants excised from in vitro raised shoots onto a medium containing cytokinins. Sucrose was the most suitable carbon source examined for shoot multiplication. An increase in shoot multiplication rate was noticed upto 4 – 5 subcultures. Nodal explants rooted on an auxin-supplemented medium. The acclimatized plants were successfully transplanted in the field. This revised version was published online in July 2006 with corrections to the Cover Date.     

In vitro system for studying the interaction between Erwinia amylovora and genotypes of pear

A new in vitro system is described for studying an interaction between Erwinia amylovoraand Pyrus communis (L.). The system uses single shoots placed onto the solid medium, and it enables to detect changes in pH of the medium and differential appearance of shoot necrosis. Shoots of susceptible cultivar (Williams) and tolerant cultivar (Harrow Sweet) were compared measuring the necrosis rate along the in vitroshoots and the pH variation following proton extrusion of both plant and pathogen. Shoots acidified differentially the culture medium depending on the presence of the pathogen, cultivar susceptibility and shoot inoculation methods. Differences in the tolerance level against pathogen among the cultivars were distinguishable only when the shoots were inoculated at the basal end. In susceptible cultivar, the necrosis appeared after 48 h of inoculation, while in tolerant cultivars after 72 h. This system is repeatable and more reliable than already known methods, such as in vitroleaf explants or in vivoplants; it can be used all around the year to test the gene expression and products essential to characterize the genes involved in the pathogenesis. This system showed the effects of E. amylovoraon the photosystem dependent system of host cells, confirmed by the effects of pathogen attack on the variation of chlorophyll a and chlorophyll b ratios and positive effects of light on the appearance of the first disease symptoms.     

Field studies on colour preferences of Ctenarytaina thysanura in Tasmanian boronia farms

Field tests on attraction of Ctenarytaina thysanura (Hemiptera: Psyllidae) adults to different coloured 30×30 cm sticky traps revealed a preference for yellow. Among the enamel colours tested, more psyllids were captured on yellow traps followed by green, then blue and least on red, cyan and magenta. Dilution of yellow enamel with 50% white (1Y: 1W) and 75% white (1Y: 3W) to produce yellow-white hues resulted in a significant decrease in psyllid capture indicating that the psyllids response to yellow was one of positive attraction and could suggest true colour discrimination. Reflectance spectra of painted surfaces of the enamel colours and also yellow to white hues indicated that psyllid capture rates were directly related to the proportion of light reflected in the 500–560 nm region. The biological basis of the observed C. thysanura response may be that yellow is the most intensely reflective colour in the general part of the spectrum for leaves which reflect most light in the 500–600 nm (peak 550 nm) range.     

Factors that affect plant regeneration from in vitro culture of immature seeds in four lentil cultivars

An efficient and simple method for plant regeneration from immature lentil seeds (Lens culinaris) is described. Immature seeds from 1 to 6 mm of four lentil cultivars were cultured in vitro on 10 different media. Culture media included different concentrations of N ⁶ -benzylaminopurine (BAP), alone or in combination with other phytohormones. After 4 weeks in culture, multiple shoot regeneration was observed using media with BAP. Immature seed size showed significant effect on shoot regeneration. Regenerated shoots (up to 4 shoots per explant on medium with Kinetin (KN) and from 5 to 20 on media with BAP) formed adventitious roots 30 days after transferring them to a medium containing 11.4 μM indole-3-acetic acid (IAA). The efficiency of the rooting medium varied depending upon the shoot-regeneration medium and the cultivar tested. The highest rooting percentage (88.9%) was obtained from regenerated shoots of the cultivar Verdina on a medium with 1 μM α-naphthaleneacetic acid (NAA). This revised version was published online in June 2006 with corrections to the Cover Date.     

Assessment of in vitro growth of apical stem sections and adventitious organogenesis to evaluate salinity tolerance in cultivated tomato

The possible use of in vitro shoot morphogenesis and shoot apex culture to evaluate salt tolerance in cultivated tomato (Lycopersicon esculentum Mill.) has been analyzed, using two cultivars with similar salt tolerance, Pera and Hellfrucht frühstamm (HF). The effect of salt on shoot regeneration was studied by culturing leaf explants on media supplemented with 0, 43, 86, 129 and 172 mM NaCl. The presence of NaCl in the regeneration media at 86 mM strongly inhibited shoot regeneration in the cultivar HF, but not in Pera. However, the substitution of NaCl by mannitol, maintaining the same water potential in the culture media, decreased the regeneration percentage in Pera but did not affect HF. Shoot apices of both cultivars were also subcultured at 6-week intervals, for 4 subcultures, at the same NaCl concentrations as used in the previous experiment, and the shoot growth, leaf and root number, rooted shoot and shoot necrosis were recorded at the end of each subculture. Root formation was the parameter most affected by salt in both cultivars, Pera being more sensitive than HF. The substitution of NaCl by mannitol significantly increased the percentage of rooted shoots in Pera after four subcultures, and slightly decreased this percentage in HF. Shoot necrosis was only observed in the last subculture at NaCl higher than 86 mM, the percentage of necrotic shoots being higher in Pera than in HF (75% and 45%, respectively). The lack of agreement between the results obtained with the in vitro tests, e.g., adventitious shoot formation and growth of apical stem sections, suggests that this approach may not be a reliable tool to evaluate salt tolerance in cultivated tomato. This revised version was published online in June 2006 with corrections to the Cover Date.     

Water use and chemical composition of ryegrass (Lolium) cultivars

Summary Significant differences in total dry matter yields of shoots and roots were found between 11 ryegrass (Lolium) cultivars grown in a glasshouse. Although shoot yield varied significantly between individual cultivars there was no overall difference between the annual and perennial cultivars; whereas for roots, the yields of the perennial plants were much smaller than those of the annual types. Water use (g H₂O g total DM^–1) also varied significantly between cultivars. However, there was no relationship between efficient water use and dry matter production.No significant differences were found in shoot composition between the cultviars for nitrogen, phosphorus, and potassium; however, concentrations of sulphur, magnesium, calcium, and sodium varied significantly. Sodium concentrations were generally higher in the annual compared to the perennial cultivars. For roots only nitrogen, phosphorus, and sulphur differed significantly between cultivars. Of the elements only calcium in the shoots was shown to be related to water use. Thus cultivars which were low users of water also had significantly lower calcium concentrations in their shoots. Water use appeared to affect the absorption of calcium by the root to a far greater extent than the transport from roots to shoot. An apparent relationship between magnesium concentration in the shoots and water use was shown to be due to the close association of magnesium with calcium in the plant.     

Effects of growth regulators and incubation period on in vitro regeneration of adventitious shoots from gerbera petioles

An effective system for in vitro regeneration of adventitious shoots from callus for the transformation or mutation of gerbera was developed. Callus was produced from petioles of the youngest 3–4 leaves detached from auxillary shoots produced in vitro. Induction medium, on which leaves were incubated over 3 or 6 days, contained 2.3 μM thidiazuron and 0.53 μM α-naphthaleneacetic acid. Explants were than transferred to one of three regeneration media with lower levels of growth regulators. Regeneration was quantified over four (4-weeks each) passages at the time of explant transfer to fresh medium. Direct shoot regeneration occurred during the first 4 weeks, and after these shoots were discarded a semi-compact organogenic callus was produced. Effectiveness of shoot regeneration depended on four criteria: the cultivar (three cultivars were tested), the sequence of passage on regeneration medium, the growth regulators in regeneration medium and the duration of the induction period. Regeneration potential from calli of all cultivars increased from the first to the fourth passage. Duration of the incubation period on induction medium (3 or 6 days) influenced regeneration to varying degrees, depending on the cultivar used and the regeneration medium contents. There were no differences between two of the regeneration media – B, containing 2.2 μM 6-benzyladenine and 0.3 μM indole-3-acetic acid and C, containing 4.4 μM 6-benzyladenine, 4.6 μM zeatin and 0.6 μM indole-3-acetic acid. Cultivar Mariola was the most productive and regenerated more than seven shoots per callus in the fourth passage. Regeneration on medium B was further evaluated on four additional gerbera cultivars. This revised version was published online in June 2006 with corrections to the Cover Date.     

Regeneration from Phylloclade Explants and Callus Cultures of Schlumbergera and Rhipsalidopsis

Phylloclade explants of Schlumbergera and Rhipsalidopsis were cultured in vitro to produce axillary and adventitious shoots. The explants of both species, taken from greenhouse-grown plants, produced only axillary shoots. There was a pronounced improvement in adventitious shoot formation in phylloclade explants of cultivar CB4 of Rhipsalidopsis by increasing numbers of subcultures of axillary shoots used as donor plants. The axillary shoots generated from the explants were either subcultured to produce successive generations of axillary shoot cultures or made into phylloclade explants and tested for adventitious shoot formation at each subculture. The duration of each subculture varied from 6 to 12 weeks. After the first subculture, sporadic adventitious shoot formation began, and after the third subculture 87% explants of cultivar CB4 produced adventitious shoots at a frequency of about 12 shoots per explant. In contrast, there was no improvement in regenerative ability in explants of cultivar Thor-Olga of Schlumbergera up to third subculture. Adventitious shoots could be produced by callus culture too. Cultivar CB4 was highly regenerative, producing as many as 10 adventitious shoots per square cm of callus. In vitro grown plantlets, when transferred to pots continued to show prolific growth.     

Genetic manipulation in cultivars of oilseed rape (Brassica napus) using Agrobacterium

Summary The response of oilseed rape cultivars to infection with Agrobacterium tumefaciens and A. rhizogenes and the possibility of regenerating genetically transformed oilseed rape plants were examined. The frequency at which Agrobacterium induced galls or hairy-roots on in vitro cultured plants ranged from 10% to 70%, depending on the cultivar. From galls induced by the tumorigenic strain T37, known to be strongly shoot inducing on tobacco, roots developed frequently. Occasionally, shoots formed and some of these produced tumour cell specific nopaline. Attempts to grow the transformed shoots into plants have so far been unsuccessful. Whole plants transformed with Ri-T-DNA, however, were regenerated. These had crinkled leaves and abundant, frequently branching roots that showed reduced geotropism, similar to previously isolated Ri T-DNA transformed tobacco and potato plants. The transformed oilseed rape plants flowered, but failed to form seeds.     

Rice cultivar evaluation for phosphorus use efficiency

Phosphorus deficiency is one of the most growth-limiting factors in acid soils in various parts of the world. The objective of this study was to screen 25 rice cultivars (Oryza sativa L.) at low, medium, and high levels of soil P. Number of tillers, root length, plant height, root dry weight and shoot dry weight were related to tissue P concentrations, P uptake and P-use efficiency. Shoot weight was found to be the plant parameter most sensitive to P deficiency. Significant cultivar differences in P use efficiency were found. Phosphorus use efficiency was higher in roots than shoots and decreased with increasing levels of soil P. Positive correlations were found among growth parameters such as plant height, tillers, root and shoot weight, and P content of roots and shoots. These results indicate selection of rice cultivars for satisfactory performance under low P availability can be carried out using shoot and root dry weight as criteria.     

Phase change modifies anthocyanin synthesis in Acer palmatum Thunb. (Japanese maple) cultivars

The potential markers of juvenility (cyanidin 3-glucoside and cyanidin 3-rutinoside) in autumn leaves of seven Acer palmatum Thunb. cultivars were investigated. Three shoot positions were marked on each cultivar—crown shoot, middle shoot, and basal shoot—and the anthocyanins were analyzed using HPLC-MS. The results showed great differences in cyanidin 3-glucoside and cyanidin 3-rutinoside concentrations among seven cultivars; moreover, significant differences in cyanidin 3-glucoside content levels were also observed among three shoot positions regardless of the cultivar analyzed. The concentration decreased basipetally and reached levels up to 52 times higher in leaves obtained from crown shoots in comparison to basal shoot leaves. Therefore, the content level of cyanidin 3-glucoside can be defined as a quantitative marker of positional effect in all the Acer palmatum Thunb. cultivars analyzed. The content level of cyanidin 3-rutinoside did not express the same positional dependence.     

Effects of genotype, explant source, and plant growth regulators on indirect shoot organogenesis in Dieffenbachia cultivars

The capacity for indirect shoot organogenesis of leaf and root explants of four Dieffenbachia cultivars were examined on a modified Murashige and Skoog (MS; Physiol Plant 15:473–495, 1962) medium supplemented with different plant growth regulators in 112 combinations. Callus formation was only observed from leaf explants on MS supplemented with 1–10 μM thidiazuron (TDZ) and 0.5–1.0 μM 2,4-dichlorophenoxyacetic acid (2,4-D) regardless of cultivars. The combination of 5 μM TDZ and 1 μM 2,4-D resulted in the greatest callus formation frequency among the four cultivars tested. Significant differences in callus and shoot formation from leaf explants were also observed among cultivars. Cultivars Camouflage, Camille, Octopus, and Star Bright produced green nodular, brown nodular, yellow friable, and green compact calli with corresponding maximum callus formation frequencies of 96%, 62%, 54%, and 52%, respectively. A maximum of 6.7 shoots/callus was observed in cv. Camouflage, followed by cvs. Camille and Star Bright at 3.7 and 3.5, respectively. Calli of cv. Octopus displayed no capacity for shoot organogenesis. Regardless of cultivar, callus formation was not observed on root explants. Regenerated shoots were successfully acclimatized in a shaded greenhouse condition with 100% survival.     

TDZ induces shoot regeneration in various Kalanchoë blossfeldiana Poelln. cultivars in the absence of auxin

In order to optimize shoot regeneration in Kalancho? blossfeldiana, leaf and internode explants of seven cultivars including one inter-specific were studied. The effects of various combinations of α-naphthalene acetic acid (NAA) (0, 0.57 M) and thidiazuron (TDZ) (0, 0.45, 4.5, 22.5, 67.5 μM) on MS medium were examined. In all cultivars shoot regeneration frequency and number of shoots per explant were enhanced by increasing TDZ concentration. Supplementing the media with NAA did not improve shoot regeneration. Maximum regeneration frequency and optimum concentration of TDZ for shoot regeneration depended significantly on the cultivar. Internode explants, but not leaf explants, of some cultivars, were able to produce adventitious shoots without treatment with growth regulator.     

Regeneration of plantlets from leaf and petiole explants of Pelargonium x hortorum

Summary The in vitro plant regeneration potential of vegetatively propagated geraniums (Pelargonium x hortorum) has been investigated. Using various combinations of growth regulators and a choice of different explants, a regeneration protocol has been developed to raise in vitro plantlets from young petiole and leaf explants from three different cultivars of geraniums. In all three cultivars, very young petiole explants exhibited a higher regeneration potential as compared with leaf explants. Regeneration efficiencies were found to be highly dependent on the cultivar, with cv. Samba showing the highest regeneration potential, followed by cvs. Yours Truly and then Sincerity. Samba also showed the highest number of shoots from both the petiole [57 shoot buds per petiole explant in the presence of 3 μM zeatin and 1 μM indole-3-acetic acid (IAA) and leaf explants (43 shoots per leaf explant with 10 μM zeatin and 2 μM IAA). Shoot buds transferred to Murashige and Skoog (MS) medium supplemented with 0.44 μM N⁶-benzyladenine and 0.11 μM IAA grew vigorously and attained 1–2 cm in length in 3–4 wk. These shoots rooted with 100% efficiency on MS basal medium, and plants developed that showed normal growth and flowering under greenhouse conditions.     

Organogenesis of Phaseolus angularis L.: high efficiency of adventitious shoot regeneration from etiolated seedlings in the presence of N6-benzylaminopurine and thidiazuron

A step-wise procedure for the regeneration of fertile plants by organogenesis from cultures of the economically important Phaseolus angularis L., cultivars: KS-6, KS-7 and KS-8 using etiolated seedlings was established. Pre-culture of 5-day old seedling explants with MS (Murashige and Skoog (1962) Physiol Plant 15:473–493) + B₅-vitamins (Gamborg et al. (1968) Exp Cell Res 50:151–158) liquid medium containing either 5.0 μM TDZ or 5.0 μM BAP under dark condition was essential for organogenesis. Bud growth and shoot multiplication were stimulated by reducing the BAP concentrations from 5.0 to 2.5 μM after 3 weeks. The maximum frequency of shoot induction was 65.2% (33.8 ± 2.54 shoots/explant) in cultivar KS-8 followed by KS-7 34.6% (23.4 ± 1.91 shoots/explant) and KS-6 30.6% (21.2 ± 2.28 shoots/explant). The multiplied buds elongated after transferring to solid MSB₅ medium supplemented with 4.0 μM GA₃, 12.5 μM AgNO₃ and 0.4 μM IBA. Up to 98% rooting efficiency of was obtained when the shoots were pulse-treated with liquid medium containing 4.5 μM IBA for 10 min. The rooted plantlets were transferred to pots in the greenhouse, where they grew, mature, flowered and bared pod normally. The efficient shoot bud induction capability was found to be cultivar dependent. All the three cultivars tested formed multiple shoots. This efficient and rapid regeneration system may also be helpful for Agrobacterium- or particle gun-mediated transformation for this important legume crop.     

Compensatory growth in shoot populations of young white pine trees

Summary White pine (Pinus strobus L.) trees have shoot populations composed of subpopulations of terminal and lateral shoots. I tested whether the subpopulations would show compensatory (increased) growth when separated from each other. Ten-year-old white pine (Pinus strobus L.) trees growing under an oak (Quercus) overstory were untreated or treated in winter by removing either all terminal, or all lateral buds (10 trees per treatment). Growth was compared between control and treated shoot subpopulations. In the 1st year, shoot-length frequency distributions were similar between control and treated subpopulations. There was significant compensatory shoot elongation (mean of 1.5 cm per shoot) in both treated subpopulations. In the 2nd year each subpopulation produced both terminal and lateral shoots. Shoot-length frequency distributions were similar, but shifted toward longer shoots in treated populations. Shoot number, mean length and total shoot length were greater in treated populations. The increased growth in treated subpopulations was due both to differences in parent shoot length and to compensatory shoot production and elongation.     

Callus,shoot and hairy root formation in vitro as affected by the sensitivity to auxin and ethylene in tomato mutants

We analyzed the impact of ethylene and auxin disturbances on callus, shoots and Agrobacterium rhizogenes-induced hairy root formation in tomato (Solanum lycopersicum L.). The auxin low-sensitivity dgt mutation showed little hairy root initiation, whereas the ethylene low-sensitivity Nr mutation did not differ from the control Micro-Tom cultivar. Micro-Tom and dgt hairy roots containing auxin sensitivity/biosynthesis rol and aux genes formed prominent callus onto media supplemented with cytokinin. Under the same conditions, Nr hairy roots did not form callus. Double mutants combining Rg1, a mutation conferring elevated shoot formation capacity, with either dgt or Nr produced explants that formed shoots with little callus proliferation. The presence of rol + aux genes in Rg1 hairy roots prevented shoot formation. Taken together, the results suggest that although ethylene does not affect hairy root induction, as auxin does, it may be necessary for auxin-induced callus formation in tomato. Moreover, excess auxin prevents shoot formation in Rg1.     

Transformation parameters enhancing T-DNA expression in kenaf (Hibiscus cannabinus)

Kenaf(Hibiscus cannabinus)is a fast growing annual with tremendous potential as a source of fiber for ropes, textiles and paper. Kenaf is an environmentally friendly crop; however, commercial production of kenaf is hindered by weed competition at the seedling stage. Herbicide resistant kenaf cultivars would reduce seedling weed competion and make growing kenaf more profitable. Factors that are important in establishing a transformation system for kenaf were examined. The influence of Agrobacterium strain, temperature, host tissue wounding, acetosyringone, virG/virE genes and host cell division on T-DNA expression in the kenaf shoot apex were investigated. Three Agrobacterium strains were tested, and A. tumefaciens LBA4404 significantly (α=0.05) yielded a high number of shoots surviving on selection medium; no shoots survived with EHA101S or Z707S. There was no significant difference (α=0.05) in transient T-DNA expression between 28 °C and 25 °C; however, shoots did not survive 16 °C or 19 °C co-cultivation temperatures. Shoot apex survival was increased significantly (α=0.05) when virulence genes and a cytokinin, TDZ, were combined. Sonicated shoots showed an increase in transient expression and shoot survival. Optimal conditions for shoot apex T-DNA transfer and expression were sonication for 5 s, co-cultivation with LBA4404 containingvirG/virEat room temperature, and 200 μmol/L acetosyringone.