Bulked AFLP analysis for the assessment of genetic diversity in white clover (Trifolium repens L.)

The use of bulked leaf samples from individual plants for amplified fragment length polymorphism (AFLP) analysis was evaluated as a tool for assessment of genetic diversity in white clover (Trifolium repens L.). Bulking of leaf samples produced slightly simpler AFLP profiles compared to the combined profiles of individual plants from the same cultivar. Approximately 90% of bands which were present in individual plants were present in bulked samples of the same cultivar. The majority of those absent were rare bands, shared by less than 25% of individual plants. Replicate bulk samples gave almost identical banding patterns, demonstrating the robustness of the bulked AFLP technique. Cluster analysis of AFLP data derived from individual plants resulted in a phenogram similar to that produced from data derived from bulked samples of the same plants. AFLP analysis of bulked samples detected significant amounts of genetic variability among 52 cultivars and accessions with genetic similarity values ranging from 0.42 to 0.92. However, cluster analysis of AFLP data only partially reflected the geographic origin of cultivars and accessions and was not congruent with cluster analysis based on variation for morphophysiological characters. Bulked AFLP analysis provides a powerful tool for rapid assessment of genetic variability in white clover and may also be used for cultivar identification. This revised version was published online in August 2006 with corrections to the Cover Date.     

In vitro androgenesis of wheat: from fundamentals to practical application

Wheat anther cultures have a history of almost 30 years and are nowemployed efficiently in many countries of the world for the developmentof doubled haploid lines for breeding. The present paper discusses keyquestions of the elaboration and perfection of the method: cytologicalaspects of in vitro androgenesis, the conditions required for theembryogenic development of microspores and the applicability of anthercultures in the Martonvásár wheat breeding programme.     

Wheat starch biosynthesis

Starch biosynthesis in plants involves the concerted action of a numberof enzymes, including ADPglucose pyrophosphorylase, starch synthases,branching enzymes and debranching enzymes. We report on the cloningand characterisation of genes encoding these enzymes from wheat and ontheir chromosomal locations. The prospects for manipulating wheat starchstructure and functionality using these genes is discussed.     

5‐Aminolevulinic Acid Activates Antioxidative Defence System and Seedling Growth in Brassica napus L. under Water‐Deficit Stress

The present study assesses the effects of 5‐aminolevulinic acid (ALA, 0, 0.1, 1 and 10 mg l^?1) on the growth of oilseed rape (Brassica napus L. cv. ZS758) seedlings under water‐deficit stress induced by polyethylene glycol (PEG 6000, 0 and ?0.3 MPa). Water‐deficit stress imposed negative effects on seedling growth by reducing shoot biomass, cotyledon water potential, chlorophyll content and non‐enzymatic antioxidants (glutathione and ascorbic acid) levels. On the other hand, water‐deficit stress enhanced the malondialdehyde (MDA) content, reactive oxygen species (ROS) production, enzymatic antioxidants activities, reduced/oxidized glutathione ratio (GSH/GSSG) and reduced/oxidized ascorbic acid (ASA/DHA) ratio in seedlings. Application of ALA at lower dosages (0.1 and 1 mg l^?1) improved shoot weight and chlorophyll contents, and decreased MDA in rape seedlings, whereas moderately higher dosage of ALA (10 mg l^?1) hampered the growth. The study also indicated that 1 mg l^?1 ALA improved chlorophyll content, but reduced MDA content and ROS production significantly under water‐deficit stress. Lower dosages of ALA (0.1 and 1 mg l^?1) also enhanced GSH/GSSG and ASA/DHA as compared to the seedlings under water‐deficit stress. The antioxidant enzymes (ascorbate peroxidase, peroxidase, catalase, glutathione reductase and superoxide dismutase) enhanced their activities remarkably with 1 mg l^?1 ALA treatment under water‐deficit stress. It was also revealed that 1 mg l^?1 ALA treatment alone induced the expression of APX, CAT and GR substantially and under water‐deficit stress conditions ALA treatment could induce the expression of POD, CAT and GR to a certain degree. These results indicated that 0.1–1 mg l^?1 ALA could enhance the water‐deficit stress tolerance of oilseed seedlings through improving the biomass accumulation, maintaining a relative high ratio of GSH/GSSG and ASA/DHA, enhancing the activities of the specific antioxidant enzymes and inducing the expression of the specific antioxidant enzyme genes.     

Copper‐Induced Oxidative Stress and Responses of the Antioxidant System in Roots of Medicago sativa

The effects of various copper (Cu) concentrations on the antioxidative system in the roots of Medicago sativa were explored. The results indicated that the Cu content of the roots reached a value of 854 μg g^?1 DW at 10 μm Cu and a value of 4415 μg g^?1 DW at 100 μm Cu, suggesting that M. sativa has better ability to tolerate and accumulate Cu than other Cu‐bioaccumulators, and is a potential plant for phytoremediation. Treatment with Cu resulted in a significant increment in the levels of H₂O₂, O₂˙^? and OH˙. The reduced form of ascorbate and glutathione reached a peak at 30 μm Cu, and was followed by a sharp depletion to a lower level than that of the control. In contrast, the levels of the oxidised forms of ascorbate and glutathione showed a progressive increment with increasing Cu concentrations, suggesting that the antioxidant system was unable to cope with Cu stress at higher Cu levels. Under the Cu concentrations tested, the activity of catalase (CAT, EC 1.11.1.6), ascorbate peroxidase (APX, EC 1.11.1.11) and glutathione reductase (GR, EC 1.6.4.2) increased at lower Cu concentrations, and then decreased, reaching a maximum at 30 μm of Cu for APX and GR, at 10 μm for CAT, whereas the activities of guaiacol peroxidase (POD, EC 1.11.1.7) were gradually increased with increasing Cu concentrations. PAGE analysis of superoxide dismutase (SOD, EC 1.1.5.1.1) revealed that one band is a Mn‐SOD and five bands are identified as Cu, Zn‐SOD, whereas Fe‐SOD isoforms were not found in the roots of alfalfa. Cu at 10–100 μm increased the intensity of constitutive isozymes of CAT, APX and POD, whereas it decreased the intensity of isozymes of glucose‐6‐phosphate dehydrogenase (G6PDH, EC 1.1.1.49) significantly. The activities of lipoxygenases (LOX, EC 1.13.11.12) were gradually augmented with increasing Cu concentrations, demonstrating that LOXs are probably involved in production of lipid hydroperoxides and superoxide anion. There was a continuous and pronounced enhancement in the activity of esterase (EST, EC 3.1.1.1) in roots treated with 10–30 Cu μm , whereas EST activity in roots exposed to above 30 μm Cu declined, suggesting that EST plays a protective role under lower Cu concentrations stress.     

Mapping chromosomal regions affecting flowering time in a spring wheat RIL population

Flowering time is an important trait for the adaptation of wheat to its target environments. To identify chromosome regions associated with flowering time in wheat, a whole genome scan was conducted with five sets of field trial data on a recombinant inbred lines (RIL) population derived from the cross of spring wheat cultivars ‘Nanda 2419’ and ‘Wangshuibai’. The identified QTLs involved seven chromosomal regions, among which QFlt.nau-1B and QFlt.nau-2B were homoeologous to QFlt.nau-1D and QFlt.nau-2D, respectively. Nanda 2419, the earlier flowering parent, contributed early flowering alleles at five of these QTLs. QFlt.nau-1B and QFlt.nau-7B had the largest effects in all trials and were mapped to the Xwmc59.2–Xbarc80 interval on chromosome 1BS and the Xgwm537–Xgwm333 interval on 7BS. Most of the mapped QTL intervals were not coincident with known vernalization response or photoperiod sensitivity loci and QFlt.nau-1B seems to be an orthologue of EpsA ^m 1. Four pairs of loci showed significant interactions across environments in determining flowering time, all of which involved QFlt.nau-1B. These findings are of significance to wheat breeding programs.     

Salt‐Induced Regulation of Some Key Antioxidant Enzymes and Physio‐Biochemical Phenomena in Five Diverse Cultivars of Turnip (Brassica rapa L.)

A greenhouse experiment was carried out to examine the differential morpho‐physiological responses of five cultivars of turnip (Brassica rapa L.) to salt stress. Five diverse cultivars of turnip (shaljum desi surakh, shaljum purple top, shaljum golden bal, neela shaljum, and peela shaljum) were subjected for 6 weeks to varying levels of NaCl, i.e. 0, 80 and 160 mm in Hoagland’s nutrient solution in sand culture. Imposition of varying levels of salt substantially decreased shoot and root fresh and dry weights, chlorophyll contents, leaf osmotic potential, relative water contents, different gas exchange attributes, total phenolics, malondialdehyde, activities of superoxide dismutase, peroxidase catalase, and leaf and root K⁺ levels while enhanced the proline contents, membrane permeability, level of H₂O₂, leaf and root Na⁺ and Cl^? and leaf Ca²⁺ in all turnip cultivars under study. Of all cultivars, peela shaljum and neela shaljum were consistently higher in their growth than the other turnip cultivars at all salt concentrations of the growth medium. Photosynthetic capacity (A) and stomatal conductance (g_s) were higher in high biomass‐producing cultivars, i.e. peela shaljum and neela shaljum, which provide to be potential selection criteria of salt tolerance in turnip. However, the regulation of antioxidant system was cultivar‐specific under saline conditions.     

Corresponding patterns of site quality,decline and tree growth in a sessile oak stand

This study focuses on two rarely studied aspects of oak decline: relations with site characteristics and effects on tree growth. The study was carried out in a 5.5 ha stand in Hungary which is strongly affected by oak decline. The nearly pure sessile oak (Quercus petraea) stand of mostly coppice origin was 90 years old at the beginning of the study. Within-stand site heterogeneity was described by the herbaceous vegetation. Four ecological site types were distinguished by the species composition of herbs, and characterized by the ecological indicator values of the species. Tree growth between 1987 and 1993 was measured, and tree vigour was estimated from visual characteristics five times between 1987 and 1993. Potential volume increment of declining trees was estimated with the growth rates of healthy trees of the same size. Volume increment loss caused by oak decline was also assessed. Significant positive relationships were found between tree vigour and tree size and between tree vigour and tree growth. The growth of seriously declining trees dropped to almost one-half of that of healthy ones. Growth reduction of living trees at the stand level amounted to 5.4%, whereas growth reduction of all trees, including those that died during the observation period, amounted to 19.9% of the potential growth. Tree size and growth were greater on better sites. A strong relationship was also found between tree vigour and site type, but sessile oak was more susceptible to decline at better sites.     

Cloning and Sequence Analysis of Lipoxygenase Gene cDNA from Cucumber Fruit （Cucumis sativus L.）

Lipoxygenases are nonheme-iron-containing dioxygenases that catalyze the hydroperoxidation of unsatrated fatty acids containing a cis, cis-1,4-pentadiene structure producing hydroperoxy acids with conjugated dienes. LOX activity has been found in a wide range of plants. Typical substrates for LOX in plants are linoleic acid and linilenic acid fatty acids. The function of various LOXs in plants is unknown, but their participation in all stages of plant growth and development has been suggested （Hildebrand, 1989; Siedow, 1991）. Some of the physiological processes in whicn lipoxygenses have been implicated include wounding （Saravitz and Siedow, 1996）, pathogen attack （Melan et al., 1993）, seed germination （Kato et al., 1992）, fruit ripening （Ferrie et al., 1994）, plant senescence （Paliyath and Droillard, 1992）. The study on the role of lipoxygenase in ripening and senescence fruit focused on tomato and strawberry. Cloning LOX gene of cucumber fruit will make us further understand the molecularaction mode of this enzyme during fxuit ripening and senescence. In this paper we isolated the partial nucleotide sequences of cucumber fiuit lipoxygenase gene and discuss the characterization of it.