Phytochrome Regulates Gibberellin Biosynthesis during Germination of Photoblastic Lettuce Seeds1

Germination of lettuce (Lactuca sativa L.) seed is regulated by phytochrome. The requirement for red light is circumvented by the application of gibberellin (GA). We have previously shown that the endogenous content of GA1, the main bioactive GA in lettuce seeds, increases after red-light treatment. To clarify which step of GA1 synthesis is regulated by phytochrome, cDNAs encoding GA 20-oxidases (Ls20ox1 and Ls20ox2, for L. sativa GA 20-oxidase) and 3β-hydroxylases (Ls3h1 and Ls3h2 for L. sativa GA 3β-hydroxylase) were isolated from lettuce seeds by reverse-transcription polymerase chain reaction. Functional analysis of recombinant proteins expressed in Escherichia coli confirmed that the Ls20ox and Ls3h encode GA 20-oxidases and 3β-hydroxylases, respectively. Northern-blot analysis showed that Ls3h1 expression was dramatically induced by red-light treatment within 2 h, and that this effect was canceled by a subsequent far-red-light treatment. Ls3h2 mRNA was not detected in seeds that had been allowed to imbibe under any light conditions. Expression of the two Ls20ox genes was induced by initial imbibition alone in the dark. The level of Ls20ox2 mRNA decreased after the red-light treatment, whereas that of Ls20ox1 was unaffected by light. These results suggest that red light promotes GA1 synthesis in lettuce seeds by inducing Ls3h1 expression via phytochrome action.

Storage protein mobilization and thiol protease up-regulation by solid matrix priming in loblolly pine (Pinus taeda) seed embryos

Experiments were conducted to investigate physiological mechanisms of solid matrix priming (SMP) on germination enhancement of loblolly pine (Pinus taeda) seeds. During SMP, osmotic potential in the embryo decreased by 0.65 MPa, concentration of crystalloid proteins decreased to 62% and concentrations of buffer soluble proteins and free amino acids increased by 22% and by 166%, respectively. Observations under an electron microscope demonstrated protein bodies in the embryo were mobilized. Inhibitor analysis indicated thiol protease was the dominant enzyme among endopiptidases to degrade the reserved proteins. A fragment of thiol protease was cloned from the primed seed embryos and it has high identities to those thiol proteases responsive to water stress. RNA get blot analysis showed a 1.5 kb thiol protease gene was up-regulated by SMP. Treatment with E64, a thiol protease inhibitor, negated SMP effects on germination performance, water potentials and protein profiles. Based on the experimental results, reserve protein mobilization induced by SMP in the embryo before radicle emergence might be one of the mechanisms to enhance germination in loblolly pine seeds.

Seed viability and dormancy mechanisms of Leucopogon melaleucoides Cunn. ex DC. (Epacridaceae)

Leucopogon melaleucoides, a flowering shrub, is desired by floricultural markets but is difficult to propagate. Seed viability was tested and dormancy mechanisms were studied to develop a commercial propagation system. Although around 56% of seed were viable, germination was completely inhibited unless the endocarp was removed. After-ripened seed (8 months after collection) germinated faster than fresh seed (2 days after collection), but germination occurred over a prolonged period (155 days). Germination of after-ripened seed was promoted with GA(3) or a commercial smoke product containing unknown plant growth regulators. All viable seed treated with GA(3) at 1000 mg L-1 had germinated after 24 days. The results suggest that both a physical and physiological dormancy mechanism occur for this species, with removal of the endocarp and pretreatment with 1000 mg L-1 GA(3) promoting complete germination of viable seed.

Fruit processing, seed viability and dormancy mechanisms of Persoonia sericea A. Cunn. ex R. Br. and P-virgata R. Br. (Proteaceae)

The morphology of the fruit and difficulties with fruit processing impose major limitations to germination of Persoonia sericea and P. virgata. The mesocarp must be removed without harming the embryo. Fermentation of fruit or manual removal of the mesocarp was effective but digestion in 32% hydrochloric acid (HCl) completely inhibited germination. The endocarp is extremely hard and therefore very difficult and time consuming to remove without damaging the seeds. The most efficient method was cracking the endocarp with pliers, followed by manual removal of seeds. Germination was completely inhibited unless at least half of the endocarp was removed. Microbial contamination of the fruit and seeds was controlled by disinfestation and germination of the seed under aseptic conditions. The results suggest that dormancy in these species is primarily due to physical restriction of the embryo by the hard endocarp.

Effect of harvest time and drying on supersweet sweet corn seed quality

A supersweet sweet corn hybrid, Pacific H5, was grown under field conditions in South-East Queensland to study the effects of harvest time and drying conditions on seed quality. Cobs were harvested at different times to obtain seed with two moisture percentage ranges (20-30% and 40-50%) and dried to 12% moisture under different combinations of drying temperatures (30 degrees C, 40 degrees C and 50 degrees C) and air velocities (1.25 m/s, 2.75 m/s and 4.30 m/s). Dried seed was stored at 30 degrees C with bimonthly monitoring of seed quality for 12 months. For standard as well as cold test germinations, statistical analysis yielded significant main effects for temperature, air velocity and harvest moisture content and significant interactions for drying temperature by harvest moisture and drying temperature by air velocity. Germination at the beginning of storage was unaffected by drying temperatures up to 40 degrees C regardless of harvest moisture but was lower at 50 degrees C for higher moisture. However, germination at the end of the storage period of 12 months was greatest for seed harvested at higher moisture and dried at temperatures up to 40 degrees C. Germination was not affected by air velocity for drying temperatures up to 40 degrees C but at 50 degrees C it generally decreased with increase in air velocity. To slow down seed deterioration during storage, it is recommended that sweet corn seed should be harvested at a higher moisture range (40-50%) and dried at 40 degrees C and 4.30 m/s air velocity. The drying temperature can be raised to 50 degrees C for seed harvested at a low moisture range (20-30%) provided the air velocity is kept low (1.25 m/s).

Burial depth and cultivation influence emergence and persistence of Phalaris paradoxa seed in an Australian sub-tropical environment

Emergence and persistence characteristics of Phalaris paradoxa seeds in no- and minimum-till situations and at different burial depths were studied in a sub-tropical environment. Three experiments were carried out using naturally shed seeds. In the first experiment, seedlings emerged from May through to September each year, although the majority of seedlings emerged in July. In the second experiment with greater seed density, cultivation in March of each year stimulated seedling emergence, altered the periodicity of emergence and accelerated the decline of seeds in the seedbank compared with plots that received no cultivation. The majority of seedlings in the cultivated plots emerged in May whereas the majority of seedlings in the undisturbed plots emerged in July. Emergence accounted for only 4-19% of the seedbank in both experiments over 2 years. Seed persistence was short in both field experiments, with less than 1% remaining 2 years after seed shed. In the third experiment, burial depth and soil disturbance significantly influenced seedling emergence and persistence of seed. Seedlings emerged most from seed mixed in the top 10 cm when subjected to annual soil disturbance, and from seed buried at 2.5 and 5.0 cm depths in undisturbed soil. Emergence was least from seed on the soil surface, and buried at 10 and 15 cm depths in undisturbed soil. Seeds persisted longest when shed onto the soil surface and persisted least when the soil was tilled. These results suggest that strategic cultivation may be a useful management tool, as it will alter the periodicity of emergence allowing use of more effective control options and will deplete the soil seedbank more rapidly.

Effect of growing site, moisture stress and seed size on viability and dormancy of Sporobolus pyramidalis (giant rats tail grass) seed

Sporobolus pyramidalis P. Beauv (giant rats tail grass) is a serious agricultural and environmental weed in tropical and subtropical areas of Australia. Infestations of this unpalatable plant reduce the productivity of pastures and the profitability of industries dependent on grazing animals. This paper reports a series of studies undertaken to assist in the development of control strategies for this species. In particular, these studies measured the viability and dormancy status of fresh seed of S. pyramidalis and the decline of dormancy with time. Variability in these characteristics was determined in seeds collected from several sites within south-east Queensland. The effect of moisture availability during the inflorescence and seed production phases on seed viability and dormancy was also determined. The dormancy of freshly collected seed from several sites ranged from 15 to 95%, but decreased to negligible levels after 4-6 months. Seeds that matured under conditions of high moisture availability were initially more dormant than seeds matured where moisture was less readily available. The proportion of viable seeds was significantly lower in smaller than larger seeds even though viability for all seed sizes exceeded 90%. This study has shown that seed of S. pyramidalis generally has high viability with a large proportion of the seed germinable soon after maturity.

Optimum harvest maturity for guayule seed

Guayule (Parthenium argentatum Gray) is a rubber-producing shrub native to the semi-arid region of north central Mexico and southwestern Texas. Timely harvest is critical to achieve maximum seed viability, vigour, and yield. The objective of this study was to investigate possible indicators of optimum seed maturity in guayule. The optimum harvest maturity time for guayule was studied by comparing quality parameters at different times after flowering. Heat units expressed as growing degree-days after flowering were calculated and related to seed development stages and quality. Seed quality at different stages of development was assessed by germination, capitulum dry mass, 1000 seed mass, and percentage of filled seeds. The maximum seed quality was recorded at 329 growing degree-days (GDD). This was 28 days from time of flowering. At this date, the moisture content of the capitulum was 48% on a wet basis and the colour was comparable to cinnamon (Code 165C) on the Royal Horticultural Society (R.H.S.) standard colour chart. Of all the parameters GDD, 1000 seed mass, and percentage of filled seeds provided a more rapid and reliable measure of optimum seed maturity. Colour identification can be used as an additional indicator. (C) 2005 Elsevier B.V. All rights reserved.

Climatic regulation of seed dormancy and emergence of diverse Malva parviflora populations from a Mediterranean-type environment

Malva parviflora L. (Malvaceae) is rapidly becoming a serious weed of Australian farming systems. An understanding of the variability of its seed behaviour is required to enable the development of integrated weed management strategies. Mature M. parviflora seeds were collected from four diverse locations in the Mediterranean-type climatic agricultural region of Western Australia. All of the seeds exhibited physical dormancy at collection; manual scarification or a period of fluctuating summer temperatures (50/20 degrees C or natural) were required to release dormancy. When scarified and germinated soon (1 month) after collection, the majority of seeds were able to germinate over a wide range of temperatures (5-37 degrees C) and had no light requirement. Germination was slower for seeds stored for 2 months than seeds stored for 2 years, suggesting the presence of shallow physiological dormancy. Seed populations from regions with similar annual rainfall exhibited similar dormancy release patterns; seeds from areas of low rainfall (337-344mm) were more responsive to fluctuating temperatures, releasing physical dormancy earlier than those from areas of high rainfall (436-444mm). After 36 months, maximum seedling emergence from soil in the field was 60%, with buried seeds producing 13-34% greater emergence than seeds on the surface. Scanning electron microscopy of the seed coat revealed structural differences in the chalazal region of permeable and impermeable seeds, suggesting the importance of this region in physical dormancy breakdown of M. parviflora seeds. The influence of rainfall during plant growth in determining dormancy release, and hence, germination and emergence timing, must be considered when developing management strategies for M. parviflora.

Assessing nut germination protocols for macadamia cultivar 'Beaumont'

A rapid rate and high percentage of macadamia nut germination, together with production of vigorous seedlings, are required by nurseries and breeding programs. Germination of nuts is typically protracted, however, and rarely reaches 100%. Many studies have been conducted into macadamia germination, but most have assessed percent germination only. This study investigated the effects of various treatments on percent germination, germination rate, and plant, shoot and root dry weights. The treatments tested were combinations of: (i) soaking or not soaking seeds in a dilute fungicide solution prior to planting; (ii) four different planting media; and (iii) leaving seed trays open or placing them inside clear plastic bags. For freshly harvested nuts, sowing in potting mix under clear plastic and without soaking produced the highest percent germination and germination rate, the largest shoots, and longest lateral roots.

Tetrazolium test for seed viability and germinability of Melocactus ernestii Vaupel subsp. ernestii and Melocactus zehntneri (Briton & Rose) Luetzelb. (Cactaceae)

Despite its ornamental value, some Melocactus species (Cactaceae) are threatened by several factors and only sexual propagation is possible. Thus, artificial seed banks are an appropriate method for their ex situ conservation. Suitable methods to germinate and to monitor viability are necessary for the seeds of these species. This work explored the use of tetrazolium test for monitoring seed viability of two Melocactus species. There was a correlation between the percentage of stained embryos, considering different cover area or tone stain, and the germination percentage. Unviable embryos did not stain. The embryos curved shape can explain the partial stain on the most of cases.

The Cratylia Mollis Seed Lectin Induces Membrane Permeability Transition In Isolated Rat Liver Mitochondria And A Cyclosporine A-insensitive Permeability Transition In Trypanosoma Cruzi Mitochondria.

Previous results provided evidence that Cratylia mollis seed lectin (Cramoll 1,4) promotes Trypanosoma cruzi epimastigotes death by necrosis via a mechanism involving plasma membrane permeabilization to Ca(2+) and mitochondrial dysfunction due to matrix Ca(2+) overload. In order to investigate the mechanism of Ca(2+) -induced mitochondrial impairment, experiments were performed analyzing the effects of this lectin on T. cruzi mitochondrial fraction and in isolated rat liver mitochondria (RLM), as a control. Confocal microscopy of T. cruzi whole cell revealed that Cramoll 1,4 binding to the plasma membrane glycoconjugates is followed by its internalization and binding to the mitochondrion. Electrical membrane potential (∆Ψm ) of T. cruzi mitochondrial fraction suspended in a reaction medium containing 10 μM Ca(2+) was significantly decreased by 50 μg/ml Cramoll 1,4 via a mechanism insensitive to cyclosporine A (CsA, membrane permeability transition (MPT) inhibitor), but sensitive to catalase or 125 mM glucose. In RLM suspended in a medium containing 10 μM Ca(2+) this lectin, at 50 μg/ml, induced increase in the rate of hydrogen peroxide release, mitochondrial swelling, and ∆Ψm disruption. All these mitochondrial alterations were sensitive to CsA, catalase, and EGTA. These results indicate that Cramoll 1, 4 leads to inner mitochondrial membrane permeabilization through Ca(2+) dependent mechanisms in both mitochondria. The sensitivity to CsA in RLM characterizes this lectin as a MPT inducer and the lack of CsA effect identifies a CsA-insensitive MPT in T. cruzi mitochondria.