Research Article
An Arabidopsis thaliana embryo arrest mutant exhibiting germination potential
- Jessica R. Kristof, Jennifer L. Coppersmith, Kyung Hong, Po-Pu Liu, Tanja M. Homrichhausen, Jing Sun, Ruth C. Martin, Hiroyuki Nonogaki
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- Published online by Cambridge University Press:
- 01 June 2008, pp. 55-65
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The ability to initiate radicle elongation, or germination potential, occurs in developing embryos before the completion of seed maturation. Green embryos after the walking-stick stage in developing Arabidopsis thaliana seeds germinate when excised from seeds and incubated in Murashige–Skoog (MS) medium containing 1% sucrose. Germination potential is not observed during early embryogenesis at the globular, heart and torpedo stages. Here, we describe an Arabidopsis mutant with embryos arrested at early stages of development, but still exhibiting germination potential. The mutant, termed embryo ball (eb), produced shrunken seeds containing round or irregularly shaped embryos that did not germinate. The round embryos excised from developing eb seeds were capable of growing a primary root with root hairs when incubated in media. In contrast, cotyledons were absent at the apical region of the eb embryos, although the apical region produced leaf-like structures with trichomes, indicating vegetative leaf identity. These observations suggested that morphological maturation was not essential for the induction of germination potential. The eb embryos exhibited partial desiccation tolerance that is characteristic of mature embryos at later stages of development, suggesting that cell maturation was also independent of morphological maturation. The eb mutant provides novel information on cell and tissue identity in developmental biology, as well as a useful tool to dissect the mechanisms underlying the induction of germination potential in developing seeds.
Expression and activity of a Kip-related protein, Zeama;KRP1, during maize germination
- Natividad de Jesús Juárez, Alfredo Mancilla, Elpidio García, Jorge M. Vázquez-Ramos
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- Published online by Cambridge University Press:
- 01 June 2008, pp. 67-75
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Plant KRP proteins are cyclin/cyclin-dependent kinase subunit (Cdk) inhibitors that share a limited homology with mammalian p27Kip1 proteins. Several KRPs have been reported in maize (Zea mays L.), of which Zeama;KRP1 was studied during maize germination. Expression of the Zeama;KRP1 gene did not vary during the 24 h germination period. A homologous antibody raised against the 13 kDa carboxy end of the Zeama;KRP1 polypeptide, a sequence containing the cyclin/Cdk inhibitory domain, indicated the existence of a 22 kDa protein in maize embryonic axes, the amount of which also remained unchanged during germination. Neither abscisic acid nor cytokinins modified the amount of protein. The purified Zeama;KRP1 polypeptide inhibited the kinase activity associated with Zeama;PCNA and Zeama;CycD2;1, and also the kinase activity in p13Suc1-pulled-down complexes. However, there were differences in the inhibition pattern during germination. Whereas kinase activity in proliferating cell nuclear antigen (PCNA) or CycD2;1 immunoprecipitates was strongly inhibited mainly during early germination, that in p13Suc1-pulled-down complexes was mainly inhibited at late times, suggesting that each protein complex is composed of different cyclins and/or Cdks.
Probability models for detecting transgenic plants
- Carlos M. Hernández-Suárez, Osval A. Montesinos-López, Graham McLaren, José Crossa
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- Published online by Cambridge University Press:
- 01 June 2008, pp. 77-89
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When detecting the adventitious presence of transgenic plants (AP), it is important to use an appropriate testing method in the laboratory. Dorfman's group testing method is effective for reducing the number of laboratory analyses, but does not consider the case where AP is diluted below the sensitivity of the analyses, which causes the rate of false negatives to increase. The objective of this study is to propose binomial and negative binomial probabilistic models for determining the required sample size (n), number of pools (g), and size of the pool (k) for detecting individuals possessing AP with a probability ≥ (1 − α) (for a small α) given: (1) pool size (k); (2) estimated proportion of individuals with AP in the population (p); (3) concentration of the trait of interest (AP) in individual seeds (w); and (4) detection limit of the test (c) (AP concentration in a pool below which it cannot be detected). The proposed models consider the different rates of false positives (δ) and false negatives (λ), and the assessment of consumer and producer risks. Results have shown that when using the negative binomial, a required sample size n can be determined that guarantees a high probability that m individuals or g pools containing AP will be found. The pools formed have an optimum size, such that one element with AP will be detected at a low cost. The negative binomial distribution should be used when it is known that the proportion of individuals with AP in the population is p < 0.1; thus, it is guaranteed that m individuals or g pools of individuals with AP will be detected with high probability.
Endozoochory by native and exotic herbivores in dry areas: consequences for germination and survival of Prosopis seeds
- Claudia M. Campos, Begoña Peco, Valeria E. Campos, Juan E. Malo, Stella M. Giannoni, Francisco Suárez
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- Published online by Cambridge University Press:
- 01 June 2008, pp. 91-100
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Mammalian herbivores can play a role in the endozoochorous dispersal of Prosopis seeds in the arid zones of Argentina, and the introduction of exotic mammals can change relevant parameters of the plant's reproductive ecology. Our specific goals were to quantify the seeds of Prosopis flexuosa, P. chilensis and P. torquata contained in the faeces of native [mara (Dolichotis patagonum), guanaco (Lama guanicoe)] and exotic [donkey (Equus asinus) and hare (Lepus europaeus)] mammals, and to determine the mortality, germination percentage and germination speed of seeds borne in faeces, compared with those collected from trees. In 2006, seeds were collected directly from trees or removed from mammal faeces. Seeds were counted and classified as apparently healthy or damaged. P. torquata and P. flexuosa seeds were found in the dung of guanaco, hare and mara, whereas only P. flexuosa seeds appeared in donkey faeces. P. chilensis seeds were only found in guanaco faeces. There was a notable relative abundance of seeds contained in small herbivore faeces in comparison to the large herbivores. The passage through the digestive tract of animals modified seed germination capacity and speed, with great variability between animal species. Guanacos had greater effects than donkeys on mortality, germination percentage and germination speed. Differences between the effects of both smaller-sized herbivores on seeds were much smaller.
Seed-bank dynamics in the invasive plant, Ambrosia artemisiifolia L.
- B. Fumanal, I. Gaudot, F. Bretagnolle
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- Published online by Cambridge University Press:
- 01 June 2008, pp. 101-114
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The seed-bank dynamics of Ambrosia artemisiifolia (Asteraceae), an annual invasive plant introduced from North America to Europe, were studied in nine French populations developing in field crops, set-asides or wastelands. The vertical distribution of seeds was studied in two different depth sections (0–5 cm and 5–20 cm), and the spatial horizontal pattern of distribution was analysed for two selected field-crop and set-aside populations. The proportions of dormant, non-dormant and dead seeds, as well as the changes in seed-bank dormancy over time, were evaluated. Natural seedling recruitment and its variation under different disturbance treatments in competitive set-asides were also studied. From 536 ± 194 to 4477 ± 717 seeds m− 2 were found in the living seed bank (0–20 cm). The majority of seeds in field crops was found in the lower soil layer, whereas the opposite pattern was observed in set-asides and wastelands. Dormant seed proportions were low (0–18%) before natural germination, but then regularly increased. The seed bank of A. artemisiifolia was spatially aggregated irrespective of the extent of soil disturbance. On average, 23 ± 12 to 292 ± 62 seedlings m− 2 were recorded, with densities lower in set-asides than in field crops and wastelands. Seedling densities were not correlated to upper seed densities except for two field crops. Soil disturbance and the removal of vegetation had positive effects on seedling recruitment in set-asides. Seed-bank dynamics appear to be crucial in the invasive success of A. artemisiifolia, and this knowledge will provide insights for the management of the species.
Gene flow hampered by low seed size of hybrids between oilseed rape and five wild relatives
- Wei Wei, Henri Darmency
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- Published online by Cambridge University Press:
- 01 June 2008, pp. 115-123
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Recent concern about gene flow from transgenic plants to weedy species has attracted much research on the fitness of their hybrids. However, no studies have been reported on the very early effects of the seed size of hybrids compared with parental plants for germination, seedling establishment and plant growth. We produced hybrids between male sterile Brassica napus L. (oilseed rape) and five weedy relatives, including Brassica juncea (L.) Czern, B. nigra (L.) Koch, B. rapa L., Hirschfeldia incana (L.) Lagrèze-Fossat and Raphanus raphanistrum L. The hybrid seeds formed between B. napus and B. rapa varied in size, while all the hybrid seeds formed with the other species were small. In a direct-seeded field experiment, small seeds of both parents and hybrids had a lower frequency of emergence and a lower seedling survival rate than large seeds, and resulted in later flowering with less biomass. However, no difference was recorded in a transplant experiment, indicating that growth in the juvenile period was sensitive to the small seed class in field conditions only. The optimum environmental conditions in the greenhouse probably homogenized the developmental differences observed at the early stage, and thus reduced the variation during subsequent growth in the field. This point has not been, but should be, considered in risk assessment of transgenic plants. The lower seedling establishment of small-seeded hybrids could hamper further gene flow.
Short Communication
A simple staining method for observation of germinated Striga seeds
- Shusheng Long, Venasius Lendzemo, Thomas W. Kuyper, Zhengsheng Kang, Horst Vierheilig, Siegrid Steinkellner
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- Published online by Cambridge University Press:
- 01 June 2008, pp. 125-129
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In vitro techniques are essential for Striga research and the development of appropriate control methods. In the laboratory, pre-screening of non-host or false-host plants of Striga for trap cropping or the screening of hosts for resistance involves visual evaluation of Striga seed germination that may be stimulated by plant parts or root exudates. This technique is presently laborious because the small Striga radicles are colourless. A number of solutions were evaluated to visualize the radicles of Striga hermonthica to obtain a reliable, simple and fast staining method yielding good contrast for visual observation, with readily available, inexpensive and minimally toxic dyes and staining solutions. The inks Brilliant Blue (Pelikan), Blue (Geha) and Brilliant Green (Pelikan), in either vinegar or lactic acid, produced radicles with excellent contrast, whereas radicles stained with Brilliant Red (Pelikan), Black (Sheaffer), Brilliant Black (Pelikan) gave good contrast. Striga radicles stained with Aniline Blue in vinegar or lactic acid, or Coomassie Brilliant Blue R250 in lactic acid, showed excellent contrast. Radicles stained with Ink Black (Sheaffer), Cotton Brown, or Rubin S in either vinegar or lactic acid also showed good contrast. With water as the diluent, only Lactophenol Blue showed excellent contrast. For health, safety and environmental concerns, availability and staining time, Blue ink in household vinegar (5% acetic acid) appears to be an excellent dye for Striga radicles and could be used in routine Striga germination assays.