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PubMed2003

[Using of selective marker gene in transgenic plants and its removal].

Selective marker gene is usually used to select transformed cells or tissue during gene transfer. However,the use of selective marker gene,especially antibiotic-resistant gene, is harmful to environment, plant development and affects multi-transformation. A new strategy that offers a approach for the elimination of those disadvantages caused by the selectable marker gene is developed. We summarized correlative marker genes used in transgenic plants and some methods of its removal.

English Abstract↗

PubMed1975

Water Conservation in Kalanchoe blossfeldiana in Relation to Carbon Dioxide Dark Fixation.

The succulent Kalanchoe blossfeldiana v. Poel. var Tom Thumb was treated on long and short photoperiods for 6 weeks during which short day plants developed thicker leaves, flowered prolifically, and exhibited extensive net dark fixation of carbon dioxide. In contrast, long day plants remained vegetative and did not develop thicker leaves or exhibit net carbon dioxide dark fixation. When examined after the photoperiodic state described, long day plants showed approximately three times more water loss over a 10-day period than short day plants. Water loss is similar during light and dark periods for short day plants but long day plants exhibited two times more water loss during the day than at night. The latter plants also lost three and one-half times more water during the light period than short day plants. The water conservation by short day plants is correlated with conditions of high carbon dioxide dark fixation and effects of its related Crassulacean acid metabolism on stomatal behavior.

Journal Article↗

PubMed2006

Factors modulating the levels of the allelochemical sorgoleone in Sorghum bicolor.

Sorgoleone is the major component of the hydrophobic root exudate of sorghum [Sorghum bicolor (L.) Moench]. The presence of this allelochemical is intrinsically linked to root growth and the development of mature root hairs. However, factors modulating root formation and the biosynthesis of sorgoleone are not well known. Sorgoleone production was independent of early stages of plant development. The optimum temperature for root growth and sorgoleone production was 30 degrees C. Seedling development and sorgoleone levels were greatly reduced at temperatures below 25 degrees C and above 35 degrees C. The level of sorgoleone was also sensitive to light, being reduced by nearly 50% upon exposure to blue light (470 nm) and by 23% with red light (670 nm). Applying mechanical pressure over developing seedlings stimulated root formation but did not affect the biosynthesis of this lipid benzoquinone. Sorgoleone production did not change in seedlings exposed to plant defense elicitors. On the other hand, sorgoleone levels increased in plants treated with a crude extract of velvetleaf (Abutilon theophrasti Medik.) root. This stimulation was not associated with increased osmotic stress, since decreases in water potential (Psi(w)) by increasing solute concentrations with sorbitol reduces sorgoleone production. Sorgoleone production appears to be constitutively expressed in young developing sorghum plants. Other than with temperature, changes in the environmental factors had either no effect or caused a reduction in sorgoleone levels. However, the stimulation observed with velvetleaf root crude extract suggests that sorghum seedlings may respond to the presence of other plants by releasing more of this allelochemical.

Benzoquinones↗

PubMed2006

Transport of radiocaesium by arbuscular mycorrhizal fungi to Medicago truncatula under in vitro conditions.

The capacity of arbuscular mycorrhizal (AM) fungi to take up and translocate radiocaesium (Cs) to their host has been shown using the root-organ culture (ROC) system. However, the absence of photosynthetic tissues, lack of a normal root hormonal balance and incomplete source-sink relationships may bias the bidirectional transfer of elements at the symbiotic interface and complicate transport studies. Accordingly, we developed a novel culture system [i.e. the Arbuscular Mycorrhizal-Plant (AM-P) in vitro culture system], where AM fungi and an autotrophic host plant develop under strict in vitro conditions. With this system, we unambiguously demonstrated the capacity of AM fungi to transport Cs. The extraradical fungal hyphae took up 21.0% of the initial supply of 134Cs. Translocation to the plant represented 83.6% of the 134Cs taken up. Distribution of 134Cs in the host plant was 89.8% in the mycorrhizal roots and 10.2% in the shoot. These results confirm that AM fungi can take up, translocate and accumulate Cs. They further demonstrate unambiguously and for the first time that Cs can be transferred from AM fungi to host tissues. These results suggest a potential involvement of AM fungi in Cs biogeochemical cycle and in plant Cs accumulation.

Biological Transport↗

PubMed2003

A study on the interplay between energy and matter transformation: the effect of elevated temperatures on the leaf morphology of Vitis vinifera var. Merlot.

This investigation explores the relationship between increased energy levels and leaf morphology. It tests the idea that the causal agent of development is the dissipation of energy into transformed matter. The energy under which leaves developed was modified by increasing temperatures in grape cordons through wrapping them in clear plastic sleeves in the early spring. At the higher temperatures, and energy levels, there was a small but statistically significant decrease in leaf size and a change in organization the leaves. The decrease in leaf size may be due to a reallocation of resources, either to greater shoot growth as a previous study demonstrated or to the appearance of more vectors of development in the leaves, i.e. the appearance of more developmental subsystems. The leaves that grew under the higher temperature regime were more complex, perhaps indicating that the grapes on those same vines may produce more complex juice, another expression of more developmental subsystems. The change in organization in these leaves that developed at higher temperature argues that the causal agent in plant development is energy dissipation and the concomitant transformation of matter, the latter expressed in the appearance of more growth vectors.

Plant Leaves↗

PubMed2005

[Iron from soil to plant products].

As an essential mineral, iron plays an important role in fundamental biological processes such as photosynthesis, respiration, nitrogen fixation and assimilation, and DNA synthesis. Iron is also a co-factor of many enzymes involved in the synthesis of plant hormones. The latter are involved in many pathways controling plant development or adaptative responses to environmental conditions. Iron reactivity with oxygen leads to its insolubility (responsible for deficiency) and potential toxicity, and complicates iron use by aerobic organisms. If plants lacked an active root system with which to acquire iron from the soil, most would experience iron deficiency and show physiological changes. In contrast, an excess of soluble iron, which can occur in flooded acidic soils, can lead to ferrous iron toxicity due to iron reactivity with reduced forms of oxygen and subsequent free radical production. An optimal iron concentration is thus required for a plant to grow and develop normally. This concentration depends on multiple regulatory mechanisms controlling iron uptake from soil by the roots, as well as iron transport and distribution to the various plant organs. Optimized seed iron content is a major biotechnological challenge identified by the World Health Organization, and it is therefore crucial to understand the underlying mechanisms. Iron delivery to seeds is tightly controlled, and depends on the nature of iron speciation in specific chelates, and their transport.

Iron↗

PubMed2001

The arabidopsis cell plate-associated dynamin-like protein, ADL1Ap, is required for multiple stages of plant growth and development.

Dynamin and dynamin-like proteins are GTP-binding proteins involved in vesicle trafficking. In soybean, a 68-kD dynamin-like protein called phragmoplastin has been shown to be associated with the cell plate in dividing cells (Gu and Verma, 1996). Five ADL1 genes encoding dynamin-like proteins related to phragmoplastin have been identified in the completed Arabidopsis genome. Here we report that ADL1Ap is associated with punctate subcellular structures and with the cell plate in dividing cells. To assess the function of ADL1Ap we utilized a reverse genetic approach to isolate three separate Arabidopsis mutant lines containing T-DNA insertions in ADL1A. Homozygous adl1A seeds were shriveled and mutant seedlings arrested soon after germination, producing only two leaf primordia and severely stunted roots. Immunoblotting revealed that ADL1Ap expression was not detectable in the mutants. Despite the loss of ADL1Ap, the mutants did not display any defects in cytokinesis, and growth of the mutant seedlings could be rescued in tissue culture by the addition of sucrose. Although these sucrose-rescued plants displayed normal vegetative growth and flowered, they set very few seeds. Thus, ADL1Ap is critical for several stages of plant development, including embryogenesis, seedling development, and reproduction. We discuss the putative role of ADL1Ap in vesicular trafficking, cytokinesis, and other aspects of plant growth.

Amino Acid Sequence↗

PubMed2000

The Arabidopsis sugar-insensitive mutants sis4 and sis5 are defective in abscisic acid synthesis and response.

Although soluble sugar levels affect many aspects of plant development and physiology, little is known about the mechanisms by which plants respond to sugar. Here we report the isolation of 13 sugar-insensitive (sis) mutants of Arabidopsis that, unlike wild-type plants, are able to form expanded cotyledons and true leaves when germinated on media containing high concentrations of glucose or sucrose. The sis4 and sis5 mutants are allelic to the ABA-biosynthesis mutant aba2 and the ABA-insensitive mutant abi4, respectively. In addition to being insensitive to glucose and sucrose, the sis4/aba2 and sis5/abi4 mutants also display decreased sensitivity to the inhibitory effects of mannose on early seedling development. Mutations in the ABI5 gene, but not mutations in the ABI1, ABI2 or ABI3 genes, also lead to weak glucose- and mannose-insensitive phenotypes. Wild-type and mutant plants show similar responses to the effects of exogenous sugar on chlorophyll and anthocyanin accumulation, indicating that the mutants are not defective in all sugar responses. These results indicate that defects in ABA metabolism and some, but not all, defects in ABA response can also alter response to exogenous sugar.

Abscisic Acid↗

PubMed1997

The consequence of peroxidase overexpression in transgenic plants on root growth and development.

Transgenic tobacco plants that overproduce the tobacco anionic peroxidase wilt upon reaching maturity, although having functional stomata and normal vascular anatomy and physiology. These plants were examined further to determine the cause for wilting, and thus better understand how the anionic peroxidase functions in plant growth and development. Shoots from young peroxidase overproducing plants were grafted onto wild-type tobacco root stock to determine if the roots could absorb and transmit sufficient water to maintain leaf turgidity. These grafted plants never wilted when grown in the greenhouse though shoot peroxidase activity remained ten-fold greater than in control plants, thus indicating that wilting is a consequence of peroxidase expression in the roots. Close examination of root systems revealed considerably less root mass in the transformed plant, primarily exhibited through a decrease in branching. At flowering, root growth rate and total root mass in transformed plants were less than 50% of control plants although shoot mass and growth rate were unchanged. This is in contrast to root growth in young seedlings where transformed plants performed equivalently to controls. Root hydraulic conductivity was measured to evaluate the effect of elevated peroxidase expression on water absorption and transport; however, no significant change in hydraulic conductivity was found in transformed plants. The consequence of anionic peroxidase overexpression on indoleacetic acid (IAA) metabolism was also examined. No significant difference in IAA levels was observed; however, root elongation in plants overexpressing peroxidase was insensitive to exogenous IAA. It can be concluded that the overexpression of the tobacco anionic peroxidase in transformed plants results in diminished root mass from fewer root branches, which contributes to the wilting phenomenon seen in these plants. Further, this developmental change in transformed plants may be a consequence of the metabolism of IAA by the anionic peroxidase.

Indoleacetic Acids↗

PubMed2003

Flower development: initiation, differentiation, and diversification.

Flowering is one of the most intensively studied processes in plant development. Despite the wide diversity in floral forms, flowers have a simple stereotypical architecture. Flowers develop from florally determined meristems. These small populations of cells proliferate to form the floral organs, including the sterile outer organs, the sepals and petals, and the inner reproductive organs, the stamens and carpels. In the past decade, analyses of key flowering genes have been carried out primarily in Arabidopsis and have provided a foundation for understanding the underlying molecular genetic mechanisms controlling different aspects of floral development. Such studies have illuminated the transcriptional cascades responsible for the regulation of these key genes, as well as how these genes effect their functions. In turn, these studies have resulted in the refinement of the original ideas of how flowers develop and have indicated the gaps in our knowledge that need to be addressed.

Arabidopsis↗

PubMed2004

Distribution and population development of Nasonovia ribisnigri (Homoptera: Aphididae) in iceberg lettuce.

A field study was conducted to determine the distribution and development of aphid Nasonovia ribisnigri (Mosley) (Homoptera: Aphididae) populations in iceberg lettuce, Lactuca sativa L. 'Salinas'. Lettuce plants were transplanted and caged individually in the field and inoculated with apterous N. ribisnigri at 0, 1, 2, 3, and 4 wk after transplanting in spring and fall 2002. Plants were harvested 15-50 d after inoculations; numbers of alates and apterous N. ribisnigri were counted or estimated on each leaf for each plant. Inoculations during all 5 wk of plant development resulted in successful colonization of lettuce heads. Results indicated that head formation did not reduce the risk of colonization by N. ribisnigri to iceberg lettuce; plants were susceptible to colonization by N. ribisnigri throughout their development. For later inoculations, N. ribisnigri populations were relatively smaller, and aphids were found mostly within the heads. For earlier inoculations, N. ribisnigri populations were larger, and within-plant distributions shifted toward frame leaves. The shift of population distributions toward frame leaves correlated significantly with increases in N. ribisnigri population density. For most inoculations, more aphids were present on wrapper leaves than on other leaves. The proportion of alates did not vary significantly with population density. Population development of N ribisnigri also correlated significantly with heat unit accumulation. Yellow sticky cards were used to monitor alates in each cage. Catches of N. ribisnigri alates on yellow sticky cards were significantly correlated with total numbers of alates as well as with total population sizes on individual lettuce plants.

Animals↗

PubMed1997

In situ localization of the putative movement protein (pr17) from potato leafroll luteovirus (PLRV) in infected and transgenic potato plants.

The potato leafroll virus (PLRV) 17-kDa protein (pr17), the putative movement protein for this phloem-limited luteovirus, was localized on ultrathin sections of leaves from PLRV-infected and transgenic potato plants. The transgenic plants expressed the entire viral genome from a full-length cDNA copy (PLRVfl) or only the gene encoding pr17 (ORF4) under the control of the cauliflower mosaic virus 35S promoter. Virus-infected and PLRVfl-transgenic plants developed symptoms typical of virus infection, whereas pr17-transgenic plants did not display symptoms or ultrastructural alterations. Immunogold electron microscopy using an anti-pr17-serum detected pr17 in plasmodesmata, in virus-induced vesicles, in mitochondria, and in chloroplasts of phloem cells, in PLRV-infected as well as PLRVfl-transgenic plants. In addition, in transgenic plants, pr17 was expressed in mesophyll cells (which are not infected by PLRV under natural conditions) and localized to the same sites as in phloem cells, except in plasmodesmata. In contrast, in pr17-transgenic plants the protein was never observed on organelles, but was almost exclusively associated with plasmodesmata of all leaf cell types, indicating that the targeting of pr17 to plasmodesmata is an intrinsic property of the protein. These results support the role of pr17 in PLRV movement.

Carrier Proteins↗

PubMed2006

Brassinosteroid confers tolerance in Arabidopsis thaliana and Brassica napus to a range of abiotic stresses.

In addition to an essential role in plant development, brassinosteroids (BRs) appear to have the ability to protect plants against various environmental stresses. However, studies confirming the ability of BRs to modulate plant responses to different environmental stresses are lacking. Earlier we had demonstrated that treatment with 24-epibrassinolide (EBR), a BR, increases the basic thermotolerance of Brassica napus and tomato seedlings [Plant Mol Biol 40:333-342, 1999]. Here we demonstrate that EBR treatment enhances seedling tolerance to drought and cold stresses in both Arabidopsis thaliana and B. napus, and helps to overcome a salt-stress-induced inhibition of seed germination. The ability of EBR to confer tolerance in plants to a variety of stresses was confirmed through analysis of expression of a subset of drought and cold stress marker genes. Transcriptional changes in these genes were more apparent in EBR-treated A. thaliana, in particular during earlier time points of stress. To see if BR is essential for the heat stress (HS) response, we made use of BR-deficient mutants. Both det2-1 and dwf4 mutants still expressed heat shock proteins (hsps) to high levels during HS, indicating that although BR augments thermotolerance in plants, it is not necessary for hsp expression during HS.

Arabidopsis↗

PubMed1999

Markers for hypersensitive response and senescence show distinct patterns of expression.

Controlled cellular suicide is an important process that can be observed in various organs during plant development. From the generation of proper sexual organs in monoecious plants to the hypersensitive response (HR) that occurs during incompatible pathogen interactions, programmed cell death (PCD) can be readily observed. Although several biochemical and morphological parameters have been described for various types of cell death in plants, the relationships existing between those different types of PCD events remain unclear. In this work, we set out to examine if two early molecular markers of HR cell death (HIN1 and HSR203J) as well as a senescence marker (SAG12) are coordinately induced during these processes. Our result indicates that although there is evidence of some cross-talk between both cell death pathways, spatial and temporal characteristics of activation for these markers during hypersensitive response and senescence are distinct. These observations indicate that these markers are relatively specific for different cell death programs. Interestingly, they also revealed that a senescence-like process seems to be triggered at the periphery of the HR necrotic lesion. This suggests that cells committed to die during the HR might release a signal able to induce senescence in the neighboring cells. This phenomenon could correspond to the establishment of a second barrier against pathogens. Lastly, we used those cell death markers to better characterize cell death induced by copper and we showed that this abiotic induced cell death presents similarities with HR cell death.

Apoptosis↗

PubMed2002

Null mutation of AtCUL1 causes arrest in early embryogenesis in Arabidopsis.

The SCF (for SKP1, Cullin/CDC53, F-box protein) ubiquitin ligase targets a number of cell cycle regulators, transcription factors, and other proteins for degradation in yeast and mammalian cells. Recent genetic studies demonstrate that plant F-box proteins are involved in auxin responses, jasmonate signaling, flower morphogenesis, photocontrol of circadian clocks, and leaf senescence, implying a large spectrum of functions for the SCF pathway in plant development. Here, we present a molecular and functional characterization of plant cullins. The Arabidopsis genome contains 11 cullin-related genes. Complementation assays revealed that AtCUL1 but not AtCUL4 can functionally complement the yeast cdc53 mutant. Arabidopsis mutants containing transfer DNA (T-DNA) insertions in the AtCUL1 gene were shown to display an arrest in early embryogenesis. Consistently, both the transcript and the protein of the AtCUL1 gene were found to accumulate in embryos. The AtCUL1 protein localized mainly in the nucleus but also weakly in the cytoplasm during interphase and colocalized with the mitotic spindle in metaphase. Our results demonstrate a critical role for the SCF ubiquitin ligase in Arabidopsis embryogenesis.

Amino Acid Sequence↗

PubMed2005

Genes encoding ADP-ribosylation factors in Arabidopsis thaliana L. Heyn.; genome analysis and antisense suppression.

Vesicle trafficking delivers proteins to intracellular and extracellular compartments, cellulose synthase to the plasma membrane, and non-cellulosic polysaccharides to the cell wall. The Arabidopsis genome potentially encodes 19 proteins with sequence similarities to ARFs (ADP-ribosylation factors) and its relatives such as ARLs (ARF-like proteins). ARFs are essential for vesicle coating and uncoating in all eukaryotic cells, while ARLs play more diverse roles. Nine proteins, six of them highly similar, are possible ARFs, three are putative ARL orthologues and the remainder were designated ARF-related proteins. The functions of the six highly similar, putative ARFs in whole plant development were probed by suppressing their expression with antisense. Antisense plants were severely stunted because cell production rate and final cell size were both reduced. Changed time-to-flowering, apical dominance, and fertility may reflect alterations to hormonal and other signalling pathways with which ARFs may interact. No gross changes in targeting or compartmentalization were seen in antisense plants containing GFP targeted to the ER and Golgi and changes in cell wall composition were limited to increases in some non-cellulosic polysaccharides and a relatively small decrease in cellulose. The reasons why these effects are less drastic than the effects on endomembranes and wall composition that are seen in short-term experiments with brefeldin A and with dominant negative ARF mutants are discussed.

ADP-Ribosylation Factors↗

PubMedDate not supplied

Growth and photosynthesis of loblolly pine (Pinus taeda) after exposure to elevated CO(2) for 19 months in the field.

To detect seasonal and long-term differences in growth and photosynthesis of loblolly pine (Pinus taeda L.) exposed to elevated CO(2) under ambient conditions of precipitation, light, temperature and nutrient availability, seedlings were planted in soil representative of an early, abandoned agricultural field and maintained for 19 months in the field either in open-top chambers providing one of three atmospheric CO(2) partial pressures (ambient, ambient +15 Pa, and ambient +30 Pa) or in unchambered control plots. An early and positive response to elevated CO(2) substantially increased total plant biomass. Peak differences in relative biomass enhancement occurred after 11 months of CO(2) treatment when biomass of plants grown at +15 and +30 Pa CO(2) was 111 and 233% greater, respectively, than that of plants grown at ambient CO(2). After 19 months, there was no significant difference in biomass between +15 Pa CO(2)-treated plants and ambient CO(2)-treated plants, whereas biomass of +30 Pa CO(2)-treated plants was 111% greater than that of ambient CO(2)-treated plants. Enhanced rates of leaf-level photosynthesis were maintained in plants in the elevated CO(2) treatments throughout the 19-month exposure period despite reductions in both leaf N concentration and ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) activity during the first 11 months of CO(2) exposure. Reductions in Rubisco activity indicated photosynthetic adjustment to elevated CO(2), but Rubisco-mediated control of photosynthesis was small. Seasonal shifts in sink strength affected photosynthetic rates, greatly magnifying the positive effects of elevated CO(2) on photosynthesis during periods of rapid plant growth. Greater carbon assimilation by the whole plant accelerated plant development and thereby stimulated new sinks for carbon through increased plant biomass, secondary branching and new leaf production. We conclude that elevated CO(2) will enhance photosynthesis and biomass accumulation in loblolly pine seedlings under high nutrient conditions; however, reductions over time in the relative biomass response of plants to elevated CO(2) complicate predictions of the eventual magnitude of carbon storage in this species under future CO(2) conditions.

Journal Article↗

PubMed2004

The transcriptome of rhizobacteria-induced systemic resistance in arabidopsis.

Plants develop an enhanced defensive capacity against a broad spectrum of plant pathogens after colonization of the roots by selected strains of nonpathogenic, fluorescent Pseudomonas spp. In Arabidopsis thaliana, this rhizobacteria-induced systemic resistance (ISR) functions independently of salicylic acid but requires responsiveness to the plant hormones jasmonic acid and ethylene. In contrast to pathogen-induced systemic acquired resistance, rhizobacteria-mediated ISR is not associated with changes in the expression of genes encoding pathogenesis-related proteins. To identify ISR-related genes, we surveyed the transcriptional response of over 8,000 Arabidopsis genes during rhizobacteria-mediated ISR. Locally in the roots, ISR-inducing Pseudomonas fluorescens WCS417r bacteria elicited a substantial change in the expression of 97 genes. However, systemically in the leaves, none of the approximately 8,000 genes tested showed a consistent change in expression in response to effective colonization of the roots by WCS417r, indicating that the onset of ISR in the leaves is not associated with detectable changes in gene expression. After challenge inoculation of WCS417r-induced plants with the bacterial leaf pathogen P. syringae pv. tomato DC3000, 81 genes showed an augmented expression pattern in ISR-expressing leaves, suggesting that these genes were primed to respond faster or more strongly upon pathogen attack. The majority of the primed genes was predicted to be regulated by jasmonic acid or ethylene signaling. Priming of pathogen-induced genes allows the plant to react more effectively to the invader encountered, which might explain the broad-spectrum action of rhizobacteria-mediated ISR.

Arabidopsis↗