Richard R. Williams

Mineral nutrition and plant morphogenesis

SummaryPlant morphogenesis in vitro can be achieved via two pathways, somatic embryogenesis or organogenesis. Relationships between the culture medium and explant leading to morphogenesis are complex and, despite extensive study, remain poorly understood. Primarily the composition and ratio of plant growth regulators are manipulated to optimize the quality and numbers of embryos or organs initiated. However, many species and varieties do not respond to this classical approach and require further optimization by the variation of other chemical or physical factors. Mineral nutrients form a significant component of culture media but are often overlooked as possible morphogenic elicitors. The combination of minerals for a particular plant species and developmental pathway are usually determined by the empirical manipulation of one or a combination of existing published formulations. Often only one medium type is used for the duration of culture even though this formulation may not be optimal for the different stages of explant growth and development. Furthermore, mineral studies have often focused on growth rather than morphogenesis with very little known of the relationships between mineral uptake and morphogenesis. This article examines the present knowledge of the main effects that mineral nutrients have on plant morphogenesis in vitro. In particular, the dynamics of nitrogen, phosphorus, and calcium supply during development are discussed.

Defect in Multiple Cell Cycle Checkpoints in Ataxia-Telangiectasia Postirradiation

The recent description of a novel gene (ATM) mutated in ataxia-telangiectasia (A-T), with homologies to genes encoding proteins involved in both G1/S and G2/M checkpoint control, points to a common defect in cell cycle control in A-T operating through the cyclin-dependent kinases. In this report we demonstrate that cyclin-dependent kinases are resistant to inhibition by ionizing radiation exposure in A-T cells, and this appears to be due to insufficient induction of WAF1. Exposure of control lymphoblastoid cells to radiation during S phase and in G2 phase causes a rapid inhibition of cyclin A-Cdk2 and cyclin B-Cdc2 activities, respectively. Irradiation led to a 5-20-fold increase in Cdk-associated WAF1 in these cells, which accounts at least in part for the decrease in cyclin-dependent kinase activity. In contrast, radiation did not inhibit any of the cyclin-dependent kinase activities in S phase or G2 phase in A-T cells at short times after irradiation nor was there any significant change in the level of Cdk-associated WAF1 compared to unirradiated cells. These results are similar to those reported previously for the G1 checkpoint and provide additional evidence for the involvement of ATM at multiple points in cell cycle regulation.

Single-site mutations in the carboxyltransferase domain of plastid acetyl-CoA carboxylase confer resistance to grass-specific herbicides

Grass weed populations resistant to aryloxyphenoxypropionate (APP) and cyclohexanedione herbicides that inhibit acetyl-CoA carboxylase (ACCase; EC 6.4.1.2) represent a major problem for sustainable agriculture. We investigated the molecular basis of resistance to ACCase-inhibiting herbicides for nine wild oat (Avena sterilis ssp. ludoviciana Durieu) populations from the northern grain-growing region of Australia. Five amino acid substitutions in plastid ACCase were correlated with herbicide resistance: Ile-1,781-Leu, Trp-1,999-Cys, Trp-2,027-Cys, Ile-2,041-Asn, and Asp-2,078-Gly (numbered according to the Alopecurus myosuroides plastid ACCase). An allele-specific PCR test was designed to determine the prevalence of these five mutations in wild oat populations suspected of harboring ACCase-related resistance with the result that, in most but not all cases, plant resistance was correlated with one (and only one) of the five mutations. We then showed, using a yeast gene-replacement system, that these single-site mutations also confer herbicide resistance to wheat plastid ACCase: Ile-1,781-Leu and Asp-2,078-Gly confer resistance to APPs and cyclohexanediones, Trp-2,027-Cys and Ile-2,041-Asn confer resistance to APPs, and Trp-1,999-Cys confers resistance only to fenoxaprop. These mutations are very likely to confer resistance to any grass weed species under selection imposed by the extensive agricultural use of the herbicides.

Comparative expressed sequence hybridization to chromosomes for tumor classification and identification of genomic regions of differential gene expression

Altered expression of genes can have phenotypic consequences in cancer development and treatment, developmental abnormalities, and differentiation processes. Here we describe a rapid approach, termed comparative expressed sequence hybridization (CESH), which gives a genome-wide view of relative expression patterns within tissues according to chromosomal location. No prior knowledge of genes or cloning is required, and minimal amounts of tissue can be used. Expression profiles are achieved in a manner similar to the identification of chromosomal imbalances by comparative genomic hybridization analysis. The approach is demonstrated to indicate a chromosomal region that harbors overexpressed genes that may be associated with a drug-resistant phenotype. In addition, known and new regions of differential gene expression in both normal tissues and tumor samples from the soft tissue sarcoma group of rhabdomyosarcoma (RMS) are indicated. These regions included 2p24; overexpression of MYCN at 2p24 was confirmed by quantitative reverse transcription–PCR for all of the alveolar RMS cases and did not necessarily correspond to genomic amplification. Evidence including region specific microarray analysis indicated that overexpression of several genes from a region may be required for detection by CESH. This evidence is consistent with clusters of functionally related genes and mechanisms that affect the expression of a number of genes at a particular genomic location. The distinctive CESH profiles demonstrated in different subtypes of RMS show potential for tumor classification.

The interaction of water flow and nutrients on aquatic plant growth

A long-term experiment was conducted to compare the effects of flowing and still water on growth, and the relationship between water flow and nutrients, in Aponogeton elongatus, a submerged aquatic macrophyte. A. elongatus plants were grown for 23 weeks with three levels of nutrition (0, 0.5 and 1g Osmocote Plus® fertiliser pot−1) in aquaria containing stirred or unstirred water. Fertilized plants grew much better than non-fertilized. The highest fertilizer level produced 29% wider leaves and 58% higher total dry weight in stirred water. Stirred water increased leaf area by 40% and tuber size by 81%, but only with the highest level of nutrition. These results suggest that this plant depends on its roots for mineral uptake, rather than from the open water, and the major limitation to growth in still water is the supply of dissolved inorganic carbon. It was the combined effects of nutrient availability and stirring that produced the strongest response in plant growth, morphology and composition. This study provides some explanation for the observations of others that these plants grow best in creeks or river systems with permanently flowing water.

Effect of different types and concentrations of plant growth retardants on Sturt's desert pea (Swainsona formosa)

Sturts desert pea (Swainsona formosa) has potential as a flowering pot plant, if the uneven and excessive vegetative growth can be controlled by the use of growth retardants. Ancymidol, chlormequat, daminozide, flurprimidol and paclobutrazol were applied to Sturts desert pea (SDP) in glasshouse experiments. Paclobutrazol (PBZ) at 10 mg a.i./plant consistently produced a compact pot plant with more lateral shoots. Flurprimidol at 0.5 and 1 mg a.i./plant reduced main shoot growth but had little effect on lateral shoots. CCC had no significant effect. Ancymidol at 10 or 25 mg a.i./plant and daminozide at 2500 and 5000 mg a.i./plant produced less attractive plants with serious symptoms of toxicity. Suppression of lateral shoot growth required higher concentrations of PBZ and flurprimidol compared to the main shoot.

Mineral uptake in tobacco leaf discs during different developmental stages of shoot organogenesis

Relationships between mineral uptake and tobacco shoot organogenesis were investigated during three morphogenic phases: phase 1, days 0–10, pre-meristem formation; phase 2, days 10–20, meristem initiation and formation; and phase 3, days 20–35, growth and differentiation of induced meristems into leafy shoots. The mineral content of both shoot-forming (SF) and non-shoot-forming (NSF) media was examined over the 35-day culture period. Both SF and NSF explants rapidly consumed iron during phase 1. Nitrate uptake in SF explants was high and independent of explant growth during phases 1 and 2, but greatest and strongly correlated with growth during phase 3. Phosphorus uptake was highest in SF explants during phases 2 and 3, and correlated with explant growth. Uptake of potassium, calcium and sulphur was strongly associated with explant growth during phase 3 whereas magnesium uptake was only poorly correlated with growth. Results from this study indicate that particular minerals may have an important role in regulating development as well as generally supporting growth.

Gene expression profiling of the cephalothorax and eyestalk in Penaeus monodon during ovarian maturation.

In crustaceans, a range of physiological processes involved in ovarian maturation occurs in organs of the cephalothorax including the hepatopancrease, mandibular and Y-organ. Additionally, reproduction is regulated by neuropeptide hormones and other proteins released from secretory sites within the eyestalk. Reproductive dysfunction in captive-reared prawns, Penaeus monodon, is believed to be due to deficiencies in these factors. In this study, we investigated the expression of gene transcripts in the cephalothorax and eyestalk from wild-caught and captive-reared animals throughout ovarian maturation using custom oligonucleotide microarray screening. We have isolated numerous transcripts that appear to be differentially expressed throughout ovarian maturation and between wild-caught and captive-reared animals. In the cephalothorax, differentially expressed genes included the 1,3-β-D-glucan-binding high-density lipoprotein, 2/3-oxoacyl-CoA thiolase and vitellogenin. In the eyestalk, these include gene transcripts that encode a protein that modulates G-protein coupled receptor activity and another that encodes an architectural transcription factor. Each may regulate the expression of reproductive neuropeptides, such as the crustacean hyperglycaemic hormone and molt-inhibiting hormone. We could not identify differentially expressed transcripts encoding known reproductive neuropeptides in the eyestalk of either wild-caught or captive-reared prawns at any ovarian maturation stage, however, this result may be attributed to low relative expression levels of these transcripts. In summary, this study provides a foundation for the study of target genes involved in regulating penaeid reproduction.

Effects of Post Harvest Drying on the Yield of Tea Tree Oil (Melaleuca alternifolia)

ABSTRACT Drying tea tree leaf on the stem increased the oil content from 5.8% to 7.4% yield v/dw. This increase is not a result of changing moisture content but appears to be due to an active form of postharvest oil uptake or production. The value to oil producers of drying and storing leaf prior to distillation is a more efficient distillation system. At present, distillation occurs as close as possible to foliage harvest with the aim of reducing any chance of oil volatilization. These results show that the need for immediate distillation of cut foliage is unnecessary, as the delay would actually improve the yield rather than reduce it. For the industry these results could change the accepted methods of production. To date each producer had to have their own distillation plant as a way of avoiding distillation delays and the need for carting wet leaf material. Since drying leaves on the stems of the tree increases the oil yield and since the removal of leaves from the stem is easily accomplished when the...

Molecular taxonomic clarification of Ptilotus exaltatus and Ptilotus nobilis (Amaranthaceae)

A complete botanical key for the genus Ptilotus R. Brown ( family Amarathaceae) has not yet been published. Identifying the 100 or more Ptilotus species using morphological characters has been difficult because plants often exhibit slight morphological differences and intermediate characteristics common to several species, subspecies, varieties and forms. Ptilotus exaltatus Nees and P. nobilis ( Lindl) F. Muell share many morphological characteristics, but are classified as different species predominantly based on inflorescence colour. The current study involved a molecular phylogenetic analysis of 14 Ptilotus species using sequence data from the internal transcribed spacer ( ITS) regions ITS 1 and ITS 2 within the 18S-26S nuclear rDNA. Of the 39 accessions analysed, all except those identified as P. exaltatus and P. nobilis clustered according to their respective species based on their morphological taxonomy. In contrast, all 18 P. exaltatus and P. nobilis accessions formed a distinct monophyletic clade with 99% bootstrap values and a low level of sequence variation ( GD=0.002). Taking into account the lack of reliable morphological characters for separating P. exaltatus and P. nobilis, together with the ITS sequence data showing little genetic divergence or genetic structure, we propose that P. exaltatus and P. nobilis are conspecific.