Propagation of Wollemi pine from tip cuttings and lower segment cuttings does not require rooting hormones

Domestication of the recently discovered and highly endangered Wollemi pine has relied almost entirely upon serial propagation of cuttings from a very small conservation collection. This study assessed the requirement for applied auxin to induce rooting in tip cuttings and lower segment cuttings of Wollemi pine. Both types of cuttings proved easy-to-root, with mean rooting of 71% for tip cuttings and 82% for lower segments. Auxin application (at 1.5, 3.0 or 8.0 g indole-3-butyric acid/L) did not accelerate root protrusion from propagation tubes or affect final rooting percentages.

In vitro propagation of Corymbia torelliana x C. citriodora (Myrtaceae) via cytokinin-free node culture

Hybrids between Corymbia torelliana (F.Muell.) K.D.Hill & L.A.S.Johnson and C. citriodora subsp. variegata (F.Muell.) A.R.Bean & M.W.McDonald are used extensively to establish forestry plantations in subtropical Australia. Methods were developed for in vitro seed germination, shoot multiplication and plantlet formation that could be used to establish in vitro and ex vitro clone banks of juvenile Corymbia hybrids. Effects of sodium hypochlorite concentration and exposure time on seed contamination and germination, and effects of cytokinin and auxin concentrations on shoot multiplication and subsequent rooting, were assessed. A two-step surface sterilisation procedure, involving 70% ethanol followed by 1% sodium hypochlorite, provided almost no contamination and at least 88% germination. A novel method of cytokinin-free node culture proved most effective for in vitro propagation. Lateral bud break of primary shoots was difficult to induce by using cytokinin, but primary shoots rooted prolifically, elongated rapidly and produced multiple nodes in the absence of exogenous cytokinin. Further multiplication was obtained either by elongating lateral shoots of nodal explants in cytokinin-free medium or by inducing organogenic callus and axillary shoot proliferation with 2.2 µm benzyladenine. Plantlets were produced using an in vitro soil-less method that provided extensive rooting in sterile propagation mixture. These methods provide a means for simultaneous laboratory storage and field-testing of clones before selection and multiplication of desired genotypes.

Micropropagation of vegetatively propagated crops: accelerating release of new cultivars and providing an important source of clean planting material

Micropropagation is unequalled for the rapid clonal propagation of improved cultivars from several Australian breeding programmes. This has been particularly true of the pineapple breeding programme, but it has also found an important role in the strawberry breeding programme where high-health mother stock is of paramount concern. In the banana and ginger industries, while access to new cultivars has been of importance, micropropagation has been adopted by the industry to ensure that planting materials are free from serious pests and diseases. Bananas can be used as planting material as early as the first generation ex vitro and is responsible for the establishment of laboratories and nurseries specializing in the production of pathogen-tested plants. The ginger industry, on the other hand, has used micropropagated plants as a source of disease and pest-free stock to establish a clean 'seed' scheme based on the production of conventional planting material.

Cellular stages of root formation, root system quality and survival of Pinus elliottii var. elliottii x P. caribaea var. hondurensis cuttings in different temperature environments.

Time to first root in cuttings varies under different environmental conditions and understanding these differences is critical for optimizing propagation of commercial forestry species. Temperature environment (15, 25, 30 or 35 +/- A 2A degrees C) had no effect on the cellular stages in root formation of the Slash x Caribbean Pine hybrid over 16 weeks as determined by histology. Initially callus cells formed in the cortex, then tracheids developed and formed primordia leading to external roots. However, speed of development followed a growth curve with the fastest development occurring at 25A degrees C and slowest at 15A degrees C with rooting percentages at week 12 of 80 and 0% respectively. Cutting survival was good in the three cooler temperature regimes (> 80%) but reduced to 59% at 35A degrees C. Root formation appeared to be dependant on the initiation of tracheids because all un-rooted cuttings had callus tissue but no tracheids, irrespective of temperature treatment and clone.

Cellular stages of root formation, root system quality and survival of Pinuselliottii var. elliottii x P. caribaea var. hondurensis cuttings in different temperature environments.

Time to first root in cuttings varies under different environmental conditions and understanding these differences is critical for optimizing propagation of commercial forestry species. Temperature environment (15, 25, 30 or 352C) had no effect on the cellular stages in root formation of the Slash * Caribbean Pine hybrid over 16 weeks as determined by histology. Initially callus cells formed in the cortex, then tracheids developed and formed primordia leading to external roots. However, speed of development followed a growth curve with the fastest development occurring at 25C and slowest at 15C with rooting percentages at week 12 of 80 and 0% respectively. Cutting survival was good in the three cooler temperature regimes (>80%) but reduced to 59% at 35C. Root formation appeared to be dependant on the initiation of tracheids because all un-rooted cuttings had callus tissue but no tracheids, irrespective of temperature treatment and clone.

Production of cuttings in response to stock plant temperature in the subtropical eucalypts, Corymbia citriodora and Eucalyptus dunnii

Propagation of subtropical eucalypts is often limited by low production of rooted cuttings in winter. This study tested whether changing the temperature of Corymbia citriodora and Eucalyptus dunnii stock plants from 28/23A degrees C (day/night) to 18/13A degrees C, 23/18A degrees C or 33/28A degrees C affected the production of cuttings by stock plants, the concentrations of Ca and other nutrients in cuttings, and the subsequent percentages of cuttings that formed roots. Optimal temperatures for shoot production were 33/28A degrees C and 28/23A degrees C, with lower temperatures reducing the number of harvested cuttings. Stock plant temperature regulated production of rooted cuttings, firstly by controlling shoot production and, secondly, by affecting the ensuing rooting percentage. Shoot production was the primary factor regulating rooted cutting production by C. citriodora, but both shoot production and root production were key determinants of rooted cutting production in E. dunnii. Effects of lower stock plant temperatures on rooting were not the result of reduced Ca concentration, but consistent relationships were found between adventitious root formation and B concentration. Average rooting percentages were low (1-15% for C. citriodora and 2-22% for E. dunnii) but rooted cutting production per stock plant (e.g. 25 for C. citriodora and 52 for E. dunnii over 14 weeks at 33/28A degrees C) was sufficient to establish clonal field tests for plantation forestry.

Variation of cutting rooting ability in cultivated and wild species of Macadamia

In Australia, macadamia trees are commonly propagated by germinating rootstock seed and grafting when seedlings reach a suitable size. The production of grafted trees is a protracted and complex process, however, propagation of macadamia via cuttings represents a simpler and faster method of multiplication. Macadamias have traditionally proven difficult to propagate from cuttings, and while recent developments in the process have improved success rates, substantial variation in rooting ability between cultivars and species has been reported. The cultivar 'Beaumont' (Macadamia integrifolia × M. tetraphylla) is commonly propagated by cutting for use as a rootstock, and is relatively easy to strike while other cultivars are more difficult. There is speculation that Hawaiian cultivars are more difficult to strike from cuttings than Australian cultivars due to species and genetic composition. In this experiment, cuttings of 32 genotypes were evaluated for rooting ability. Each genotype's species profile was estimated using historical data, and used to determine species effects on survival (percentage) and rooting ability (rating 0-2). M. jansenii (100%), M. tetraphylla (84%) and M. integrifolia/tetraphylla hybrids (79%) had the highest success rates while M. integrifolia (54%) and M. ternifolia (43%) had the lowest survival. Rooting ability of M. jansenii (1.75) was significantly higher than M. ternifolia (0.49) but not significantly higher than M. tetraphylla × M. integrifolia with (1.09), M. tetraphylla (1.03) or M. integrifolia (0.88).

The meleagrid herpesvirus 1 genome is partially resistant to transposition

The propagation of herpesvirus genomes as infectious bacterial artificial chromosomes (iBAC) has enabled the application of highly efficient strategies to investigate gene function across the genome. One of these strategies, transposition, has been used successfully on a number of herpesvirus iBACs to generate libraries of gene disruption mutants. Gene deletion studies aimed at determining the dispensable gene repertoire of the Meleagrid herpesvirus 1 (MeHV-1) genome to enhance the utility of this virus as a vaccine vector have been conducted in this report. A MeHV-1 iBAC was used in combination with the Tn5 and MuA transposition systems in an attempt to generate MeHV-1 gene interruption libraries. However, these studies demonstrated that Tn5 transposition events into the MeHV-1 genome occurred at unexpectedly low frequencies. Furthermore, characterization of genomic locations of the rare Tn5 transposon insertion events indicated a nonrandom distribution within the viral genome, with seven of the 24 insertions occurring within the gene encoding infected cell protein 4. Although insertion events with the MuA system occurred at higher frequency compared with the Tn5 system, fewer insertion events were generated than has previously been reported with this system. The characterization and distribution of these MeHV-1 iBAC transposed mutants is discussed at both the nucleotide and genomic level, and the properties of the MeHV-1 genome that could influence transposition frequency are discussed. © American Association of Avian Pathologists.