Drought-Induced Changes in Protein Patterns of Brassica napus var. oleifera Roots

Drought-induced changes in two-dimensional silver stained protein patterns of Brassica napus L. var. oleifera M. root system were detected both at quantitative and qualitative levels. Particularly, 13 new polypeptides of low molecular weight were evidenced in the drought-stressed tap root, 12 of which were also present in the short tuberized roots, a specific drought-induced root type. The reversibility of these modifications, observed after 3 days rehydration, suggests that they might be involved in drought tolerance.     

Cold-deacclimation of oilseed rape (Brassica napus var. oleifera) in response to fluctuating temperatures and photoperiod

The aim of this work was to establish the role of factors that may trigger elongation growth in the dehardening response, namely temperature during daylight, photoperiod and vernalization. Fully cold-acclimated seedlings of winter (with incomplete vernalization) and spring oilseed rape were subjected to deacclimation under temperatures of 2/12, 12/2, 12/12, 12/20, 20/12 and 20/20 degrees C (day/night) and a 12 h photoperiod. Plants were also deacclimated under photoperiods of 8 and 16 h at constant temperatures of 12 and 20 degrees C. After deacclimation, plants were subjected to reacclimation. Results suggest that the level of growth activity induced during deacclimation affects both the deacclimation rate and the capacity for reacclimation. Deacclimation is fully reversible if it is not accompanied by induction of elongation growth. In such cases the rate of the decrease in freezing tolerance depends on the mean temperature of deacclimation. Deacclimation becomes partially or completely irreversible when it is connected with promotion of elongation growth. The stimuli triggering elongation growth during deacclimation may be the growth-promoting temperature (20 degrees C) during the day and the lack of vernalization blockage of elongation growth. When elongation growth was stimulated by other factors such as long-day treatments, rehardening was also disturbed.     

Drought adaptability of Agrobacterium rhizogenes-induced roots in oilseed rape (Brassica napus var. oleifera)

Abstract. Soil grown oilseed rape ( Brassica napus L. var. oleifera M., cv. Darmor) seedlings at the cotyledon stage (one week old), were inoculated in vivo at the base of the hypocotyl with Agrobacterium rhizogenes harbouring the pRi 15834 plasmid. Resulting adventitious root formation was observable about 2 or 3 weeks after infection. Differential Ri-induced root emergence and subsequent development occurred depending on water conditions and closeness of the wounding site to the soil surface: either thin, hairy roots growing rapidly and plagiotropically at the soil level under humid atmosphere, or hairless and fleshy, slowly growing aerial roots developed. The hairy roots were highly drought susceptible, whereas aerial roots revealed some potential for drought tolerance. Unlike normal roots, none of these Ri-induced roots appeared able to give rise to drought rhizogenesis in plants subjected to progressive drought stress. However, under hardening, achieved through successive and moderate drought stress-rehydration cycles, both types of Ri-induced roots improved drought tolerance and could express the morphogenetic differentiation programme leading to the formation of short, tuberized, drought-adapted, roots. These results, discussed in terms of hormonal imbalance and drought tolerance regulation, suggest that the Ri T-DNA gene expression, responsible for adventitious root induction and growth behaviour, is further regulated through the host plant.     

Changes in Root Bacterial Communities Associated to Two Different Development Stages of Canola (Brassica napus L. var oleifera) Evaluated through Next-Generation Sequencing Technology

Crop production may benefit from plant growth-promoting bacteria. The knowledge on bacterial communities is indispensable in agricultural systems that intend to apply beneficial bacteria to improve plant health and production of crops such as canola. In this work, the diversity of root bacterial communities associated to two different developmental phases of canola (Brassica napus L.) plants was assessed through the application of new generation sequencing technology. Total bacterial DNA was extracted from root samples from two different growth states of canola (rosette and flowering). It could be shown how bacterial communities inside the roots changed with the growing stage of the canola plants. There were differences in the abundance of the genera, family, and even the phyla identified for each sample. While in both root samples Proteobacteria was the most common phylum, at the rosette stage, the most common bacteria belonged to the family Pseudomonadaceae and the genus Pseudomonas, and in the flowering stage, the Xanthomonadaceae family and the genus Xanthomonas dominated the community. This implies in a switch in the predominant bacteria in the different developmental stages of the plant, suggesting that the plant itself interferes with the associated microbial community.     

Stable isotope carbon study: long-term partitioning during progressive drought stress in Brassica napus var. oleifera

Abstract. Long-term carbon partitioning was analysed by stable carbon isotope labelling of CO₂ during the adaptive response of Brassica napus to progressive drought stress. This method allowed us to distinguish between the pre-existing carbon, which had accumulated before the change in CO₂ isotope composition (on 20 d) and the recent photosynthetic input occurring during the following period. Three successive adaptive phases characterized the plant response to drought. In the first period of water shortage (20–30 d), growth was progressively slowed down: the recent C input (28.3 mg per plant) was mainly allocated to mature leaves (14 mg) and roots (8.9 mg); the pre-existing C chains were partly lost by respiration (12.1 mg) and partly translocated to the apex (2.3 mg). The increase in dry matter (13.0 mg) was mainly due to the recent C input (19.1 mg) in shoots but the roots appeared also as an important sink despite their small dry matter increase (3.2 mg). Root dry matter turnover corresponded to the elimination of pre-existing C chains (6.0 mg) and their renewal with the recent C input (9.2 mg). Root sink strength was related also to the development of drought-induced short tuberized roots (0.1 mg from pre-existing C and 0.3 mg from recent C). In the second period of water stress (from 30 to 40 d), the whole plant biomass remained constant in spite of efficient allocations from pre-existing and recent C sources to two main sinks: shoot apex (3.1 and 5.4 mg, respectively) and short tuberized roots (1.0 and 0.2 mg, respectively). The hypocotyl acted as a transient storage organ for the recent C input (2.0 mg). In the third period of recovery upon rehydration (from 46 to 51 d), the short tuberized roots gave rise to a new root system using C chains from pre-existing and recent sources (1.7 and 2.8 mg, respectively). Such concomitant sink-source behaviour of tuberized roots underlines the adaptive value of drought rhizogenesis.     

甘蓝型油菜与蓝花子远缘杂交及双二倍体的合成研究

甘蓝型油菜品种奥罗×蓝花子杂种Ｆ-１,平均配对构型为１２．１Ⅰ＋６．５３Ⅱ＋０．４１Ⅲ＋０．１８Ⅳ＋０．１８Ⅴ,Ａ、Ｃ染色体组与Ｒ染色体间存在配对,它们之间具有一定的同源性。在甘蓝型油菜与蓝花子的杂种Ｆ-１代中,存在一种染色体不配对的减数分裂类型。这一类型中有少量可形成平衡的不减数配子。提供了油菜与蓝花子远缘杂种回交结实的细胞学根据。在ＭＳ＋０．２ｍｇ／ＬＮＡＡ＋３ｍｇ／ＬＢＡ＋１ｇ／Ｌ秋水仙碱＋３０ｇ／Ｌ蔗糖＋８ｇ／Ｌ琼脂的培养基中,接种甘蓝型油菜奥罗与蓝花子的杂种Ｆ-１进行加倍处理,经快速繁殖后,获得大量的染色体数为２ｎ＝５６的双二倍体幼苗。上述双二倍体自交结实,在减数分裂中绝大多数细胞形成２８个二价体,个别形成２６个二价体和１个四价体。上述技术在油菜与蓝花子远缘杂交中首次解决了用常规方法不易获取远种杂种稳定双二倍体的难题。甘蓝型油菜品种Ａｌｔｅｘ×蓝花子杂种Ｆ-１代,长时间的快速繁殖后,出现了染色体丢失和加倍,有形成１９条染色体的配子回复到甘蓝型油菜染色体组成的趋势。在油菜远缘杂种中发现了类似于球茎大麦远缘杂种中染色体丢失的现象。     

Comparative Impacts of Heavy Metals on Root Growth as Related to Their Specificity and Selectivity

Two-day-old maize (Zea mays L.) seedlings were incubated on the solutions of Ag, Cd, Pb, Zn, Cu, Tl, Co, and Hg salts (0.001 to 3 g/l). Toxicity of heavy metals was assessed as the inhibition of root growth on the first, second, and third days, the change in the length of the lateral root zone, and the duration of lateral root development from the first division in pericycle to emergence. For all salts under study, the ratio of the lethal concentration to the lowest concentration slowing down root growth was about ten, and growth inhibition was not almost enhanced in the course of three days. With concentrations calculated as g/l, metal toxicity declined in the following order: Cu Tl > Ag > Cd > Hg > Co > Zn > Pb; for molar concentrations, the order was the following: Tl³⁺ > Cu²⁺ > > Ag⁺ > Hg²⁺ Cd²⁺ > Zn²⁺ Pb²⁺ Co²⁺. Duration of lateral root development was least affected by heavy metals. Metal affinity of biological compounds for SH-groups was closely correlated (r = 0.9) with the molar concentration that inhibited primary root growth by 50%. Because of the narrow range of effective concentrations, only slightly increasing inhibition over the exposure time, tolerant root branching, and close relationship between the toxicity and the constant of binding to SH-groups, we conclude that the salts under study exert nonselective inhibition and root growth is slowed down due to the general toxicity of heavy metals rather than selective inhibition of any particular process or processes.     

Specificity and Photomorphogenic Nature of Ultraviolet-B-Induced Cotyledon Curling in Brassica napus L

Three general classes of photomorphogenic photoreceptors have been characterized in higher plants: phytochrome, a blue light/ultraviolet (UV)-A photoreceptor(s), and a UV-B sensory system(s). Although a great deal is known about phytochrome and the blue light/UV-A photoreceptor(s), little is known about UV-B detection processes. One reason for this is the lack of readily quantifiable morphogenic responses that are specifically induced by UV-B radiation. We have discovered a response to UV-B, upward curling of Brassica napus L. cotyledons, that may be useful for probing the mechanism of UV-B photoreception. The process was initially observed when B. napus seeds were germinated under visible light plus UV-B radiation, but did not occur under visible light alone or visible light plus UV-A. When 5-d-old seedlings grown in visible light were given relatively short exposures of UV-B (100 min of 5.5 [mu]mol m-2 s-1), the curling response was also observed. Development of curling was separated from the application of this UV-B pulse by a 14-h latent period. Pulses of red light, blue light, farred light, and UV-A (100 min of 5.5 [mu]mol m-2 s-1) did not induce curling, indicating UV-B specificity Additionally, these other spectral regions did not reverse or enhance the UV-B-triggered response. The degree of curling showed a log-linear dependence on UV-B fluence (6-40 mmol m-2) and reciprocity with respect to length of exposure and fluence rate. The data indicate that curling is photomorphogenic in nature and may be triggered by a single photoreceptor species.     

Al Partitioning Patterns and Root Growth as Related to Al Sensitivity and Al Tolerance in Wheat

Studies of Al partitioning and accumulation and of the effect of Al on the growth of intact wheat (Triticum aestivum L.) roots of cultivars that show differential Al sensitivity were conducted. The effects of various Al concentrations on root growth and Al accumulation in the tissue were followed for 24 h. At low external Al concentrations, Al accumulation in the root tips was low and root growth was either unaffected or stimulated. Calculations based on regression analysis of growth and Al accumulation in the root tips predicted that 50% root growth inhibition in the Al-tolerant cv Atlas 66 would be attained when the Al concentrations were 105 [mu]M in the nutrient solution and 376.7 [mu]g Al g-1 dry weight in the tissue. In contrast, in the Al-sensitive cv Tam 105, 50% root growth inhibition would be attained when the Al concentrations were 11 [mu]M in the nutrient solution and 546.2 [mu]g Al g-1 dry weight in the tissue. The data support the hypotheses that differential Al sensitivity correlates with differential Al accumulation in the growing root tissue, and that mechanisms of Al tolerance may be based on strategies to exclude Al from the root meristems.     

Identification of Candidate Genes Related to Stem Development in Brassica napus Using RNA-Seq

Plant Molecular Biology Reporter - Plant stems are involved in supporting the entire plant body, thus having an important effect on the yield of oilseed rape. The current understanding of the...     

Differential responses of Brassica napus and Petunia hybrida to leaf protoplast isolation stress

Changes in the response to abiotic stress during the isolation of leaf protoplasts were compared between a recalcitrant species of Brassica napus and regenerating species of Petunia hybrida . Initially, levels of soluble free putrescine (put), spermidine (spd) and spermine (spm) in leaves and protoplasts were determined. The sum of these three polyamines increased in petunia and B. napus leaf protoplasts by 1.6-fold and 1.1-fold, respectively. The soluble free fraction of spd and spm decreased in B. napus but not in petunia protoplasts. During the isolation of leaf protoplasts from B. napus , the ratio of soluble free put to the total PAs almost doubled, but that of spd and spm declined significantly. Petunia leaf protoplasts treated with cyclohexylamine (CHA), an inhibitor of spermidine synthase, accumulated ammonia and soluble putrescine, but lost the soluble spermidine. The soluble polyamine levels of CHA-treated petunia leaf protoplasts corresponded with those in B. napus . Leaves were subjected to abiotic stress during the isolation of protoplasts, namely wounding and osmotic stress which changed soluble free polyamine levels in B. napus and petunia, respectively. Both B. napus and petunia leaf protoplasts showed an increase in ammonia, but total free amino acid content and activation of proteases were only enhanced in B. napus leaf protoplasts. These results suggest that in B. napus wounding initiated senescence of leaf protoplasts during their isolation, leading to a constant production of ethylene early in the culture.     

Partial Root Drying Effects on Biomass Production in Brassica napus and the Significance of Root Responses

Partial root drying (PRD) has been shown to stimulate stomatal-closure response and improve water-use efficiency and thus biomass production and grain yield under water deficiency. While most studies focus on above-ground responses to PRD, we examined how root responses contributed to effects of partial root drying. In particular, in two experiments with oilseed rape (Brassica napus L.) we investigated whether roots were able to forage for patchily distributed water, and how this affected plant growth compared with uniform watering and alternate watering (in which different parts of the roots receive water alternately). The first pot experiment was carried out in the greenhouse and the second outside under a rain-shelter in which also the watering amount was varied. The results indicate that B. napus roots were able to forage for fixed water patches by selective root placement. In the first experiment with small plants, root foraging was equally effective as enhanced water-use efficiency under alternate watering. Both treatments resulted in about 10% higher shoot biomass compared with uniform watering. Alternate watering generally outperformed uniform watering in the second experiment, but the success depended on the time of harvest and the water supply level. Measurements indicated that only the alternate watering regime effectively reduced stomatal conductance, but lead to a higher shoot biomass only under more severe (50%) rather than under milder water deficiency (70% of a well watered control). Water deficiency strongly reduced leaf initiation rates and leaf sizes in B. napus, but for a given level of water supply the supply pattern (uniform control, fixed patchy or alternate watering) hardly influenced these growth parameters. Although also in the second experiment, the plants selectively placed their roots in the wet parts of the pot, root foraging was not as effective as in the first experiment. Possible reasons for these discrepancies are discussed as well as their implications for the application of PRD effects for crop growth.     

Low temperature-induced modifications in cell ultrastructure and localization of phenolics in winter oilseed rape (Brassica napus L. var. oleifera L.) leaves

Acclimation of winter oilseed plants in the cold (i.e. at temperatures >0 degrees C) followed by short exposure to sub-lethal freezing temperatures resulted in pronounced ultrastructural changes of leaf epidermal and mesophyll cells. The following major changes were observed upon acclimation at 2 degrees C: increased thickness of cell walls; numerous invaginations of plasma membranes; the appearance of many large vesicles localized in the cytoplasm in close proximity to the central vacuole; the occurrence of abundant populations of microvesicles associated with the endoplasmic reticulum (ER) cisternae or located in the vicinity of dictyosomes; and the occurrence of paramural bodies and myelin-like structures. In addition, large phenolic deposits were observed in the vicinity of the plasma membrane and membrane-bound organelles such as chloroplasts, large vesicles or cytoplasm/tonoplast interfaces. Transient freezing (-5 degrees C for 18 h) of the cold-acclimated leaves led to reversible disorganization of the cytoplasm and to pronounced structural changes of the cellular organelles. Chloroplasts were swollen, with the stroma occupying one half of their volume and the thylakoid system being displaced to the other half. Large phenolic aggregates disappeared but distinct layers of phenolic deposits were associated with mitochondrial membranes and with chloroplast envelopes. In frost-thawed cells recovered at 2 degrees C for 24 h, dictyosomes and dictyosome- or ER-derived small vesicles reappeared in the ribosome-rich cytoplasm. Aberrations in the structure of chloroplasts and mitochondria were less pronounced. Few phenolic deposits were seen as small grains associated with chloroplast envelopes and vesicle membranes. These observations demonstrate that plants undergo different changes in cell ultrastructure depending on whether they are subjected to chilling or freezing temperatures. Results are discussed in relation to membrane recycling and the possible role of phenolics during the first and second stages of plant acclimation at low temperature.     

Resistance to Water Absorption in Germinating Rapeseed (Brassica napus L.)

Dynamics of the imbibition phase of rapeseed (Brassica napusL.) was studied by evaluating water content-water potentialrelations and the permeability to water. It was found that resistanceto water absorption was very high (diffusivity was very low)during the early stages of water absorption. As seed water contentincreased, resistance decreased by more than six orders of magnitude. The concept of hydraulic conductivity of a seed was introduced.The change in hydraulic conductivity with change in seed watercontent was similar to that observed for diffusivity. It wasshown that at late stages of water absorption hydraulic conductivityapproached values found in soils at the dry end of the availablewater range. For this reason it was suggested that hydraulicconductivity in soil may have an influence on imbibition ratein the late stages of water absorption.     

Changes in the composition of fatty acids and sterols of membrane lipids during induction and differentiation of Brassica napus (var. oleifera L.) callus

Changes in the membrane lipid and sterols content and composition were studied during induction and differentiation in callus cultures of Brassica napus var. oleifera. Callus induction was associated with an increase of DGDG content, significant changes in fatty acids composition of all lipid fractions and increased degree of lipid unsaturation. The membrane lipid composition of tissue at different degrees of differentiation was found to vary significantly, particularly two weeks after transfer of callus to regeneration medium. The main differences concerned the content and composition of galactolipids. Curiously in many cases, these differences declined during subsequent culture, in spite of the morphogenesis process which was in progress. Another result of differentiation was the change in free sterol composition: in shoot regenerating calli the content of stigmasterol had rose whereas the accumulation of campesterol decreased. Even though observed changes in membrane properties may not play a role in morphogenesis they are nevertheless useful as developmental markers and can be invaluable in understanding biochemical basis of morphogenesis.     

Low Temperature Affects Pattern of Leaf Growth and Structure of Cell Walls in Winter Oilseed Rape (Brassica napus L., var. oleifera L.)

Three-week acclimation of winter oilseed rape (Brassica napusL. var. oleifera L.) plants in the cold (2 °C) resultedin a modified pattern of leaf cell enlargement, indicated bythe increased thickness of young leaf blades and modified dimensionsof mesophyll cells, as compared with non-acclimated tissuesgrown at 20/15 °C (day/night). The thickness of leaf cellwalls also increased markedly during cold acclimation but itdecreased in response to a transient freezing event (5 °Cfor 18 h followed by 6 or 24 h at 2 °C, in the dark). Cellwalls of the upper (adaxial) epidermis were most affected. Theirultrastructure was modified by cold and freezing treatmentsin different ways, as revealed by electron microscopy. Possiblereasons for the cold- and freezing-induced modifications inthe leaf and cell wall morphology and their role in plant acclimationto low temperature conditions are discussed. Copyright 1999Annals of Botany Company Acclimation, Brassica napus var. oleifera, cell wall ultrastructure, cold, freezing, leaf structure, winter oilseed rape.     

The Use of Ergosterol as a Quantitative Measure of the Resistance of Cultured Tissues of Brassica napus ssp. oleifera to Leptosphaeria maculans

A simplified method for the quantitative assessment of the fungal lipid ergosterol was used to assess the levels of infection in tissue cultures of oilseed rape (Brassica napus ssp. oleifera) inoculated with Leptosphaeria maculans. The growth of L. maculans in liquid culture throughout a 36-day period correlated well (r = 0·92) with the amount of ergosterol extracted from the mycelium. There were significant differences (P < 0·05) in the amount of ergosterol extracted from infected thin cell layer (TCL) explants and callus tissue of two resistant and three susceptible cultivars of oilseed rape. Amounts of ergosterol extracted from resistant cultivars were < 100 (g and from susceptible > 100 (g. The mean amounts of ergosterol extracted from shoot cultures of two resistant and four susceptible cultivars were similar to those for TCL explants and callus tissue, although the values obtained were variable. This technique can be used in in vitro breeding programmes to accurately assess the resistance of tissue cultures of B. napus to L. maculans and could also have value in conventional breeding programmes.