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.

Domestication for Conservation of an Endangered Species: The Case of the Wollemi Pine

A small population of tall slender conifers was discovered in 1994 in a deep rainforest canyon of the Wollemi National Park, New SouthWales, Australia. The living trees closely resembled fossils that were more than 65 million years old, and this ‘living fossil’ was recognised as a third extant genus in the Araucariaceae (Araucaria, Agathis and now Wollemia). The species was named the Wollemi pine (W. nobilis). Extensive searches uncovered very few populations, with the total number of adult trees being less than 100. Ex situ collections were quickly established in Sydney as part of the Wollemi Pine Recovery Plan. The majority of the ex situ population was later transferred to our custom-built facility in Queensland for commercial multiplication. Domestication has relied very heavily on the species’ amenability to vegetative propagation because seed collection from the natural populations is dangerous, expensive, and undesirable for conservation reasons. Early propagation success was poor, with only about 25% of cuttings producing roots. However, small increases in propagation success have a very large impact on a domestication program because plant production can be modelled on an exponential curve where each rooted cutting develops into a mother plant that, in turn, provides more rooted cuttings. An extensive research program elevated rooting percentages to greater than 80% and also provided in vitro methods for plant multiplication. These successes have enabled international release of the Wollemi pine as a new and attractive species for ornamental horticulture.

Comparative seed and dispersal ecology of three exotic subtropical Asparagus species

The genus Asparagus includes at least six invasive species in Australia. Asparagus aethiopicus and A. africanus are invasive in subtropical Australia, and a third species, A. virgatus is naturalized and demonstrates localized spread in south east Queensland. To better understand how the attributes of these species contribute to their invasiveness, we compared fruit and seed traits, germination, seedling emergence, seed survival, and time-to-maturity. We further investigated dispersal ecology of A. africanus, examining the diet of a local frugivore, the figbird (Sphecotheres viridis) and the effect of gut passage on seedling emergence. Overall, A. aethiopicus was superior in germination and emergence, with the highest mean germination (98.8%) and emergence (94.5%) under optimal conditions and higher emergence (mean of 73.3%) across all treatments. In contrast, A. africanus had the lowest germination under optimal conditions (71.7%) and low mean seedling emergence (49.5%), but had fruits with the highest relative yield (ratio of dry pulp to fruit fresh weight) that were favored by a local frugivore. Figbirds consumed large numbers of A. africanus fruits (~30% of all non-Ficus fruits), and seedling germination was not significantly affected by gut passage compared to unprocessed fruits. Asparagus virgatus germinated poorly under cool, light conditions (1.4%) despite a high optimum mean (95.0%) and had low mean performance across emergence treatments (36.3%). The species also had fruits with a low pulp return for frugivores. For all species, seed survival declined rapidly in the first 12 mo and fell to < 3.2% viability at 36 mo. On the basis of the traits considered, A. virgatus is unlikely to have the invasive potential of its congeners. Uniformly short seed survival times suggest that weed managers do not have to contend with a substantial persistent soil-stored seed bank, but frugivore-mediated dispersal beyond existing infestations will present a considerable management challenge.

Seed persistence of the invasive aquatic plant, Gymnocoronis spilanthoides (Asteraceae)

Seed persistence of Gymnocoronis spilanthoides (D.Don) DC.; Asteraceae (Senegal tea), a serious weed of freshwater habitats, was examined in relation to burial status and different soil moisture regimes over a 3-year period. Seeds were found to be highly persistent, especially when buried. At the end of the experiment, 42.0%, 27.3% and 61.4% of buried seeds were viable following maintenance at field capacity, water logged and fluctuating (cycles of 1 week at field capacity followed by 3 weeks’ drying down) soil moisture conditions, respectively. Comparable viability values for surface-situated seeds were ~3% over all soil moisture regimes. Predicted times to1% viability are 16.2 years for buried seed and 3.8 years for surface-situated seed. Persistence was attributed primarily to the absence of light, a near-obligate requirement for germination in this species, although secondary dormancy was induced in some seeds. Previous work has demonstrated low fecundity in field populations of G. spilanthoides, which suggests that soil seed banks may not be particularly large. However, high levels of seed persistence, combined with ostensibly effective dispersal mechanisms, indicate that this weed may prove a difficult target for regional or state-wide eradication.

Cynodon dactylon, Green Couch Grass, Bermuda Grass 'Grand Prix'

‘Grand Prix’ is a selection from a cross between ‘Wintergreen’ and ‘Couch 5’ (also designated C5). ‘Couch 5’ was a selection from an earlier series of crosses by the breeder between ‘Wintergreen’ and a number of Cynodon dactylon accessions, which were collected by the breeder from the Mornington Peninsula area of Victoria between 1986 and 1990. C5 was an experimental breeding line, and was not subsequently reserved as vegetative germplasm. Living material of C5 is no longer in existence. Following the crossing of ‘Couch 5’ and ‘Wintergreen’ in 1998, the resultant seed was germinated on moist blotting paper. Individual seedlings, a total of 150 in number, were planted into 150mm pots and these plants observed during 1998 and 1999. During the summer of 1999-2000, the majority of the seedling plants were culled on the basis of their shoot density, leaf texture, internode length, and colour. In the spring of 2000, the remaining 20 potted seedlings were planted individually into 4m2 plots at the Evergreen Turf farm at Pakenham (Victoria), and allowed to expand fully across these plots. The final selection of Seedling 12 (later designated DN12) in late 2002 was based on shoot density, leaf colour, turf quality, and reduced thatch accumulation as expressed in these plots. Propagation: the original plant has been multiplied through four (4) vegetative expansions prior to PBR application without showing any discernible off types. Breeder: David Nickson, Frankston, VIC. PBR Certificate Number 3133, Application Number 2005/291, granted 12 September 2006.

Cynodon dactylon, Green Couch Grass, Bermuda Grass 'Winter Gem'

‘Winter Gem’ is a selection from a cross between ‘Wintergreen’ and Couch 5 (also designated C5). Couch 5 was a selection from an earlier series of crosses by the breeder between ‘Wintergreen’ and a number of Cynodon dactylon accessions, which were collected by the breeder from the Mornington Peninsula area of Victoria between 1986 and 1990. C5 was an experimental breeding line, and was not subsequently reserved as vegetative germplasm. Living material of C5 is no longer in existence. Following the crossing of Couch 5 and ‘Wintergreen’ in 1998, the resultant seed was germinated on moist blotting paper. Individual seedlings, a total of 150 in number, were planted into 150mm pots and these plants observed during 1998 and 1999. During the summer of 1999-2000, the majority of the seedling plants were culled on the basis of their shoot density, leaf texture, internode length, and colour. In the spring of 2000, the remaining 20 potted seedlings were planted individually into 4m2 plots at the Evergreen Turf farm at Pakenham (Victoria), and allowed to expand fully across these plots. The final selection of Seedling 9 (later designated DN9) in late 2002 was based on shoot density, leaf texture, and retention of winter colour as expressed in these plots. Propagation: The original plant had been multiplied through four (4) vegetative expansions prior to PBR application without showing any discernible off types. Breeder: David Nickson, Frankston, VIC. PBR Certificate Number 3132, Application Number 2005/290, granted 11 September 2006.

Cynodon transvaalensis x Cynodon dactylon, Hybrid Green Couch Grass, Hybrid Bermuda Grass 'P18'

‘P18’ was first produced in 1992 and is a mutant genotype obtained from a hybrid Bermudagrass line believed to be ‘Tifdwarf’, which was grown in a greenhouse owned by H&H Seed Company in Yuma, Arizona. ‘P18’ was selected for its extremely fine leaf texture, its high shoot density under close mowing, its rapid growth rate, and its uniform dark green colour, and was subsequently evaluated for these traits and characteristics. Propagation: vegetative. Breeder: Howard E. Kaewer, Eden Prairie, MN, USA. PBR Application Number 2007/179, Certificate Number 3567, granted 13 August 2007.

Power system stability implications from electromechanical wave propagation

Electromechanical wave propagation characterizes the first-swing dynamic response in a spatially delayed manner. This paper investigates the characteristics of this phenomenon in two-dimensional and one-dimensional power systems. In 2-D systems, the wave front expands as a ripple in a pond. In 1-D systems, the wave front is more concentrated, retains most of its magnitude, and travels like a pulse on a string. This large wave front is more impactful upon any weak link and easily causes transient instability in 1-D systems. The initial disturbance injects both high and low frequency components, but the lumped nature of realistic systems only permits the lower frequency components to propagate through. The kinetic energy split at a junction is equal to the generator inertia ratio in each branch in an idealized continuum system. This prediction is approximately valid in a realistic power system. These insights can enhance understanding and control of the traveling waves.

The effect of foliar sprays of the synthetic auxin 3-5-6 TPA, on fruit drop, fruit size and seed development in three lychee (Litchi chinensis) cultivars: Tai So, Fay Zee Sui and Kwai Mai Pink.

Fruit drop in lychee can cause major yield losses in Australia, the severity varying with cultivar and season. Research in China, South Africa and Israel has demonstrated the potential for synthetic auxins used as foliar sprays to reduce fruit drop in lychee. Trials were initiated in Australia to test the efficacy of the synthetic auxin, 3-5-6 Trichloro-2-phridyl-oxyacetic acid (3-5-6 TPA) at 50 ppm on the cultivars Tai So, Fay Zee Sui and Kwai Mai Pink. Results indicate that in most cases the TPA reduced natural fruit drop however the size of the fruit at the time of application affects the response and the ideal application time varies with cultivar; approximately 13 mm fruit length in 'Kwai Mai Pink', 20 mm in 'Fay Zee Sui' and 27 mm in 'Tai So'. If applied too early in 'Tai So', it caused an increase in fruit drop. The TPA was most effective when natural fruit drop was high, reducing fruit drop from 74.7 to 34.9% in 'Kwai Mai Pink' and least effective when natural fruit drop was low. An increase in the percentage of fruit with poorly developed (chicken tongue) seed and slightly larger fruit size was also observed in treated trees.

Emergence and seed persistence of Echinochloa colona, Urochloa panicoides and Hibiscus trionum in the sub-tropical environment of north-eastern Australia.

Better understanding of seed-bank dynamics of Echinochloa colona, Urochloa panicoides and Hibiscus trionum, major crop weeds in sub-tropical Australia, was needed to improve weed control. Emergence patterns and seed persistence were investigated, with viable seeds sown at different depths in large in-ground pots. Seedlings of all species emerged between October and March when mean soil temperatures were 21-23C. However, E. colona emerged as a series of flushes predominantly in the first year, with most seedlings emerging from 0-2 cm. Urochloa panicoides emerged mostly as a single large flush in the first two years, with most seedlings emerging from 5 cm. Hibiscus trionum emerged as a series of flushes over three seasons, initially with majority from 5 cm and then 0-2 cm in the later seasons. Longevity of the grass seed was short, with <5% remaining after burial at 0-2 cm for 24 months. In contrast, 38% of H. trionum seeds remained viable after the same period. Persistence of all species increased significantly with burial depth. These data highlight that management strategies need to be tailored for each species, particularly relating to the need for monitoring, application times for control tactics, impact of tillage, and time needed to reduce the seed-bank to low numbers.

Wet-dry cycling extends seed persistence by re-instating antioxidant capacity

Seeds in the field experience wet-dry cycling that is akin to the well-studied commercial process of seed priming in which seeds are hydrated and then re-dried to standardise their germination characteristics. To investigate whether the persistence (defined as in situ longevity) and antioxidant capacity of seeds are influenced by wet-dry cycling, seeds of the global agronomic weed Avena sterilis ssp. ludoviciana were subjected to (1) controlled ageing at 60% relative humidity and 53.5°C for 31 days, (2) controlled ageing then priming, or (3) ageing in the field in three soils for 21 months. Changes in seed viability (total germination), mean germination time, seedling vigour (mean seedling length), and the concentrations of the glutathione (GSH) / glutathione disulphide (GSSG) redox couple were recorded over time. As controlled-aged seeds lost viability, GSH levels declined and the relative proportion of GSSG contributing to total glutathione increased, indicative of a failing antioxidant capacity. Subjecting seeds that were aged under controlled conditions to a wet-dry cycle (to −1 MPa) prevented viability loss and increased GSH levels. Field-aged seeds that underwent numerous wet-dry cycles due to natural rainfall maintained high viability and high GSH levels. Thus wet-dry cycles in the field may enhance seed longevity and persistence coincident with re-synthesis of protective compounds such as GSH.

Seed production of Stylosanthes guyanensis. 1. Growth and development.

The pattern of growth and development of seed crops of stylo (Stylosanthes guyanensis) was derived from measurements made on experimental and commercial crops in north Queensland. The three cultivars Cook, Endeavour, and Schofield differed appreciably only in the timetable of their development. Each had distinct successive phases of vegetative and reproductive development culminating in total annual seed production of 700-800 kg ha-1 from a healthy closed canopy, the main recorded cause of reduced production being the disease Botrytis sp. In a healthy crop of Cook, the peak quantity of standing seed represented almost 90 per cent of the total accountable seed, and the rise to and decline from this peak proceeded at rates of the order of 3-4 per cent per day. It is deduced that, although there appears to be little potential for either increase in overall production or improvement in synchronization or retention characteristics beyond that currently attained by a closed canopy of healthy plants, there is scope for an increase in the efficiency of recovery of standing seed. Maximum recovery will be achieved through attention to choice of time of harvest, presentation of a minimum amount of extraneous vegetation to the harvester, and improvement in harvester separation.

Seed production of Stylosanthes guyanensis. 2. The consequences of defoliation.

The consequences of defoliation on seed production of stylo (Stylosanthes guyanensis) were examined in field experiments at Walkamin in north Queensland. The practical aim of defoliation is to present a level uncompacted crop canopy to the harvester without a reduction in the quantity of seed carried at harvest ripeness. It was concluded that the latest date at which defoliation is reasonably certain to achieve its objectives is about four weeks before first flower initiation. In north Queensland, this means late February for cvv. Cook and Endeavour and early April for cv. Schofield. The results suggest that development of the population of individual shoots must be synchronized to produce the highest peaks of standing seed; that this is best achieved by ensuring that a closed crop canopy with a ceiling shoot population exists at the time of first flower initiation; and that poorly synchronized shoot development is a consequence of defoliating too late and a cause of reduced seed production.

Seed production of Stylosanthes guyanensis. 3. Effects of pre-harvest desiccation.

Strips within commercial crops of Stylosanthes guyanensis in the Mareeba district of north Queensland were sprayed with diquat 4, 6 and 10 days before harvest and compared with unsprayed strips. Pre-harvest desiccation made combine harvesting easier, but did not increase harvest yield. Where seed formation and maturation was still possible, desiccation prevented this without substantially increasing the loss of seed to the ground; increased harvest efficiency was thus offset by a diminished quantity of standing seed. However, where there was little or no further potential for seed development, diquat had virtually no effect on the quantity of standing seed or harvest efficiency. It was concluded that the results warranted neither recommendation nor further evaluation of preharvest desiccation of S. guyanensis seed crops.

Effects of stage of defoliation on seed production and growth of Stylosanthes humilis.

Stylosanthes humilis swards grown at Brisbane in irrigated boxes were defoliated (about 60 per cent removal of tops) at floral initiation, first flower appearance, or advanced flowering stages ; seed yield was 45, 16, and 14 per cent respectively of seed yield in undefoliated swards. Decreased yields were primarily due to poor seed set of florets, were also associated with reduced inflorescence density and floret number per inflorescence, and occurred despite increases (in some defoliation treatments) in seed size, leaf growth rate, and differentiation of leaves and branches. Total seasonal plant growth was independent of defoliation treatment.