Genetic diversity among 45 Indian mustard (Brassica juncea L.) genotypes comprising 37 germplasm collections, five advance breeding lines and three improved cultivars was investigated at the DNA level using the random amplified polymorphic DNA (RAPD) technique. Fifteen primers used generated a total of 92 RAPD fragments, of which 81 (88%) were polymorphic. Of these, 13 were unique to accession ‘Pak85559’. Each primer produced four to nine amplified products with an average of 6.13 bands per primer. Based on pairwise comparisons of RAPD amplification products, Nei and Li's similarity coefficients were calculated to evaluate the relationships among the accessions. Pairwise similarity indices were higher among the oilseed accessions and cultivars showing narrow ranges of 0.77–0.99. An unweighted pair-group method with arithmetic averages cluster analysis based on these genetic similarities placed most of the collections and oilseed cultivars close to each other, showing a low level of polymorphism between the accessions used. However, the clusters formed by oilseed collections and cultivars were comparatively distinct from that of advanced breeding lines. Genetically, all of the accessions were classified into a few major groups and a number of individual accessions. Advanced breeding lines were relatively divergent from the rest of the accessions and formed independent clusters. Clustering of the accessions did not show any pattern of association between the RAPD markers and the collection sites. A low level of genetic variability of oilseed mustard was attributed to the selection for similar traits and horticultural uses. Perhaps close parentage of these accessions further contributed towards their little diversity. The study demonstrated that RAPD is a simple and fast technique to compare the genetic relationship and pattern of variation among the gene pool of this crop.

The effect of thermal stress on the antioxidant system was investigated in two invasive plants, Eupatorium adenophorum Spreng. and E. odoratum L. The former is sensitive to high temperature, whereas the latter is sensitive to low temperature. Our aim was to explore the relationship between the response of antioxidant enzymes and temperature in the two invasive weeds with different distribution patterns in China. Plants were transferred from glasshouse to growth chambers at a constant 25 °C for 1 week to acclimatize to the environment. For the heat treatments, temperature was increased stepwise to 30, 35, 38 and finally to 42 °C. For the cold treatments, temperature was decreased stepwise to 20, 15, 10 and finally to 5 °C. Plants were kept in the growth chambers for 24 h at each temperature step. In E. adenophorum, the coordinated increase of the activities of antioxidant enzymes was effective in protecting the plant from the accumulation of active oxygen species (AOS) at low temperature, but the activities of catalase (CAT), guaiacol peroxidase (POD), ascorbate peroxidase (APX), glutathione reductase (GR), and monodehydroascorbate reductase (MDAR) were not accompanied by the increase of superoxide dismutase (SOD) during the heat treatments. As a result, the level of lipid peroxidation in E. adenophorum was higher under heat stress than under cold stress. In E. odoratum, however, the lesser degree of membrane damage, as indicated by low monodehydroascorbate content, and the coordinated increase of the oxygen. Detoxifying enzymes were observed in heat-treated plants, but the antioxidant enzymes were unable to operate in cold stress. This indicates that the plants have a higher capacity for scavenging oxygen radicals in heat stress than in cold stress. The different responses of antioxidant enzymes may be one of the possible mechanisms of the differences in temperature sensitivities of the two plant species.

Wetlands are important for the protection of water quality of rivers and lakes, especially those adjacent to agricultural landscapes, by intercepting and removing nutrients in runoff. In this study, the 15N tracer technique was applied to study the distribution and fate of anthropogenic nitrogen (15N-fertilizer) in Calamagrostis angustifolia Kom wetland plant-soil microcosms to identify the main ecological effects of it. 15NH415NO3 solution (14.93 mg N/L, 20.28 at.% 15N) was added to each microcosm of the first group, which was approximate to the current nitrogen concentration (CNC) of farm drainage, and 29.86 mg N/L 15NH415NO3 solution was added to another group, which was approximate to the double nitrogen concentration (DNC) of farm drainage, while no nitrogen (NN) was added to the third group. The results suggest that the input of anthropogenic nitrogen has positive effects on the biomass and total nitrogen content of plant, and the positive effects will be elevated as the increase of its input amount. The increase of 15N-fertilizer can also elevate its amounts and proportions in plant nitrogen. Soil nitrogen is still the main source of plant nitrogen, but its proportion will be reduced as the increase of 15N-fertilizer. The study of the fate of 15N-fertilizer indicates that, in CNC treatment, only a small proportion is water-dissolved (0.13 ± 0.20%), a considerable proportion is soil-immobilized (17.02 ± 8.62%), or plant-assimilated (23.70 ± 0.92%), and most is lost by gaseous forms (59.15 ± 8.35%). While in DNC treatment, about 0.09 ± 0.15% is water-dissolved, 15.33 ± 7.46% is soil-immobilized, 23.55 ± 2.86% is plant-assimilated, and 61.01 ± 5.59% is lost by gaseous forms. The double input of anthropogenic nitrogen can not elevate the proportions of plant-assimilation, soil-immobilization and water-dissolution, but it can enhance the gaseous losses.

To investigate the sensitive site of antioxidant systems in chloroplast under cadmium stress and its consequence on reactive oxygen species production and action, the sub-organellar localization of chloroplast superoxide dismutases (SOD, EC 1.15.1.1) and ascorbic peroxidase (APX, EC 1.11.1.11) isoenzymes and changes of enzymes activities under cadmium stress were investigated in tomato seedlings. Two APX isoforms, one thylakoid-bound and one stromal, were detected. Cd at 50 μM induced a moderate increase of SOD activities but a rapid inactivation of both APX isoenzymes. APX inactivation was mainly related to the decrease of ascorbate concentration, as supported by in vitro treatment of exogenous ascorbate and APX kinetic properties under Cd stress. H2O2 accumulation in chloroplast, as a consequence of APX inactivation, was associated with a 60% loss of Rubisco (EC 4.1.1.39) activity, which could be partially accounted for by a 10% loss of Rubisco content. Protein oxidation assay found that the Rubisco large subunit was the most prominent carbonylated protein; the level of carbonylated Rubisco large subunit increased fivefold after Cd exposure. Thiol groups in the Rubisco large subunit were oxidized, as indicated by non-reducing electrophoresis. Treating crude extract with H2O2 resulted in a similar pattern of protein oxidation and thiols oxidation with that observed in Cd-treated plants. Our study indicates that APXs in the chloroplast is a highly sensitive site of antioxidant systems under Cd stress, and the inactivation of APX could be mainly responsible for oxidative modification to Rubisco and subsequent decrease in its activity.

Phytohormones play critical roles in regulating plant responses to stress. We investigated the effects of water stress induced by adding 12% (w/v) polyethylene glycol to the root medium on the levels of abscisic acid (ABA), indole-3-acid (IAA), zeatin (ZT), and gibberellin3 (GA3) in maize leaves. The results suggested that water stress had significant effects on the four hormone levels. There was a transient increase in the IAA content during the initial stage of adaptation to water stress in maize leaves, but it dropped sharply thereafter in response to water stress. ABA content increased dramatically in maize leaves after 24 h of exposure to water stress, and then the high levels of ABA were maintained to the end. The contents of ZT and GA3 rapidly declined in maize leaves subjected to water stress. The effects of water stress on chlorophyll content, electrolyte leakage and malondialdehyde levels in maize leaves were also studied. The variation of cell damage was negatively correlated with ZT and GA3 levels in maize leaves under water stress. Thus, we explored the roles of ZT and GA3 on the growth of maize seedlings under water stress by exogenous application. It is possible that both ZT and GA3 were effective in protecting maize seedlings from water stress, which would be of great importance for the improvement of drought tolerance in maize by genetic manipulation.

Nitric oxide (NO) has been known as an important signal in plant antioxidative defense but its production and roles in water stress are less known. The present study investigated whether NO dependence on a NO synthase-like (NOS) activity is involved in the signaling of drought-induced protective responses in maize seedlings. NOS activity, rate of NO release and drought responses were analyzed when NO donor sodium nitroprusside (SNP), NO scavenger c-PTIO (2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide) and NOS inhibitor L-NAME (NG-nitro-L-arginine methyl ester) were applied to both detached maize leaves and whole plants. Both NOS activity and the rate of NO release increased substantially under dehydration stress. The high NOS activity induced by c-PTIO as NO scavenger and NO accumulation inhibited by NOS inhibitor L-NAME in dehydration-treated maize seedlings indicated that most NO production under water deficit stress may be generated from NOS-like activity. After dehydration stress for 3 h, detached maize leaves pretreated with NO donor SNP maintained more water content than that of control leaves pretreated with water. This result was consistent with the decrease in the transpiration rate of SNP-treated leaves subjected to drought treatment for 3 h. Membrane permeability, a cell injury index, was lower in SNP-treated maize leaves under dehydration stress for 4 h when compared with the control leaves. Also, superoxide dismutase (SOD) activity of SNP combined drought treatment maize leaves was higher than that of drought treatment alone, indicating that exogenous NO treatment alleviated the water loss and oxidative damage of maize leaves under water deficit stress. When c-PTIO as a specific NO scavenger was applied, the effects of applied SNP were overridden. Treatment with L-NAME on leaves also led to higher membrane permeability, higher transpiration rate and lower SOD activities than those of control leaves, indicating that NOS-like activity was involved in the antioxidative defense under water stress. These results suggested that NO dependence on NOS-like activity serves as a signaling component in the induction of protective responses and is associated with drought tolerance in maize seedlings.

Rice has a preference for uptake of ammonium over nitrate and can use ammonium-N efficiently. Consequently, transporters mediating ammonium uptake have been extensively studied, but nitrate transporters have been largely ignored. Recently, some reports have shown that rice also has high capacity to acquire nitrate from growth medium, so understanding the nitrate transport system in rice roots is very important for improving N use efficiency in rice. The present study identified four putative NRT2 and two putative NAR2 genes that encode components of the high-affinity nitrate transport system (HATS) in the rice (Oryza sativa L. subsp. japonica cv. Nipponbare) genome. OsNRT2.1 and OsNRT2.2 share an identical coding region sequence, and their deduced proteins are closely related to those from mono-cotyledonous plants. The two NAR2 proteins are closely related to those from mono-cotyledonous plants as well. However, OsNRT2.3 and OsNRT2.4 are more closely related to Arabidopsis NRT2 proteins. Relative quantitative reverse transcription-polymerase chain reaction analysis showed that all of the six genes were rapidly upregulated and then downregulated in the roots of N-starved rice plants after they were re-supplied with 0.2 mM nitrate, but the response to nitrate differed among gene members. The results from phylogenetic tree, gene structure and expression analysis implied the divergent roles for the individual members of the rice NRT2 and NAR2 families. High-affinity nitrate influx rates associated with nitrate induction in rice roots were investigated and were found to be regulated by external pH. Compared with the nitrate influx rates at pH 6.5, alkaline pH (pH 8.0) inhibited nitrate influx, and acidic pH (pH 5.0) enhanced the nitrate influx in 1 h nitrate induced roots, but did not significantly affect that in 4 to 8 h nitrate induced roots.

Genomic in situ hybridization banding (GISH-banding), a technique slightly modified from conventional GISH, was used to probe the Chinese native rye (Secale cereale L.) DNA, and enabled us to visualize the individual rye chromosomes and create a universal reference karyotype of the S. cereale chromosome 1R to 7R. The GISH-banding approach used in the present study was able to discriminate S. cereale chromosomes or segments in the wheat (Triticum aestivum L.) background, including the Triticale, wheat-rye addition and translocation lines. Moreover, the GISH-banding pattern of S. cereale subsp. Afghanicum chromosomes was consistent with that of Chinese native rye cv. Jingzhou rye; whereas the GISH-banding pattern of Secale vavilovii was different from that of S. cereale, indicating that GISH-banding can be used to study evolutionary polymorphism in species or subspecies of Secale. In addition, the production and application of GISH-banding to the study of adenine-thymine-riched heterochromatin is discussed.

Wheat grown in Mn-deficient soil has been widely observed to produce much reduced yields. Breeding for Mn-efficient wheat genotypes adapted to Mn-deficient soils would represent a long-term solution for wheat agronomy. To characterize the physiological basis of Mn efficiency in wheat genotypes would facilitate the breeding programs for producing Mn-efficient wheat. Using a solution culture and a soil culture system in the present study, a Mn-efficient UK wheat genotype Maris Butler and a Mn-inefficient UK wheat genotype Paragon have been compared with a Mn-efficient Australian wheat genotype C8MM in the responses to Mn deficiency in order to characterize the Mn efficiency in these wheat genotypes. Results showed that in solution culture, Maris Butler grown under Mn deficiency had 77% relative dry matter yield of control plants that were grown under Mn sufficiency, whereas C8MM and Paragon had 60% and 58% relative dry matter yield of their respective controls. Results from the soil culture demonstrated that relative dry matter yield remained high for Maris Butler and C8MM (53% and 56%, respectively), whereas the value for Paragon dropped to 33%. In terms of dry matter yield and photosynthetic efficiency, Maris Butler demonstrated Mn efficiency in both solution culture and soil culture, whereas C8MM showed Mn efficiency only in soil culture. Results also demonstrated that under Mn-depleted supply in soil, plants of C8MM had a significantly higher ability in Mn uptake, whereas plants of Maris Butler showed a higher internal Mn use efficiency in comparison with plants of Paragon. Results from the present study indicate that the ability of C8MM to accumulate higher amounts of Mn is the basis of the improved Mn efficiency of this genotype in comparison with Paragon, and in Maris Butler there is a higher internal use of Mn expressed as an improved photosynthetic efficiency in conferring its Mn efficiency. It is suggested that more than one mechanism has arisen in wheat to confer tolerance to Mn deficiency.

Cell-wall invertase plays an important role in sucrose partitioning between source and sink organs in higher plants. To investigate the role of cell-wall invertases for seed development in rice (Oryza sativa L.), cDNAs of three putative cell-wall invertase genes OsCIN1, OsCIN2 and OsCIN3 were isolated. Semi-quantitative reverse transcription-polymerase chain reaction analysis revealed different expression patterns of the three genes in various rice tissues/organs. In developing caryopses, they exhibited similar temporal expression patterns, expressed highly at the early and middle grain filling stages and gradually declined to low levels afterward. However, the spatial expression patterns of them were very different, with OsCIN1 primarily expressed in the caryopsis coat, OsCIN2 in embryo and endosperm, and OsCIN3 in embryo. Further RNA in situ hybridization analysis revealed that a strong signal of OsCIN2 mRNA was detected in the vascular parenchyma surrounding the xylem of the chalazal vein and the aleurone layer, whereas OsCIN3 transcript was strongly detected in the vascular parenchyma surrounding the phloem of the chalazal vein, cross-cells, the aleurone layer and the nucellar tissue. These data indicate that the three cell-wall invertase genes play complementary/synergetic roles in assimilate unloading during the grain filling stage. In addition, the cell type-specific expression patterns of OsCIN3 in source leaf blades and anthers were also investigated, and its corresponding physiological roles were discussed.

Lentil, one of the oldest legumes was investigated for diversity based on botanical descriptors, total seed proteins, isozymes and random amplified polymorphic DNA (RAPD) markers. About one fourth of accessions were heterogeneous for botanical descriptors and a seed protein profile. The germplasm collected from the province of Baluchistan revealed the prevalence of indigenous landraces as high diversity was observed for all of the techniques. Diversity explored through various techniques revealed validity irrespective of the sample size or geographic pattern, RAPD being the best choice for investigating both inter- and intra-accession variation in lentil. Although all of the techniques were able to resolve genetic diversity in lentil, isozymes and seed proteins gave low levels of genetic diversity, suggesting that more investigation into isozymes of specific proteins is required. RAPD is the best option for determining inter- and intra-accession variation, and will be required to extend germplasms and primers to continue the study of botanical descriptors.

This study introduces the construction of the first intraspecific genetic linkage map of the A-genome diploid cotton with newly developed simple sequence repeat (SSR) markers using 189 F2 plants derived from the cross of two Asiatic cotton cultivars (Gossypium arboreum L.) Jianglingzhongmian × Zhejiangxiaoshanlüshu. Polymorphisms between the two parents were detected using 6 092 pairs of SSR primers. Two-hundred and sixty-eight pairs of SSR primers with better polymorphisms were picked out to analyze the F2 population. In total, 320 polymorphic bands were generated and used to construct a linkage map with JoinMap3.0. Two-hundred and sixty-seven loci, including three phenotypic traits were mapped at a logarithms of odds ratio (LOD) ≥ 3.0 on 13 linkage groups. The total length of the map was 2 508.71 cM, and the average distance between adjacent markers was 9.40 cM. Chromosome assignments were according to the association of linkages with our backbone tetraploid specific map using the 89 similar SSR loci. Comparisons among the 13 suites of orthologous linkage groups revealed that the A-genome chromosomes are largely collinear with the At and Dt sub-genome chromosomes. Chromosomes associated with inversions suggested that allopolyploidization was accompanied by homologous chromosomal rearrangement. The inter-chromosomal duplicated loci supply molecular evidence that the A-genome diploid Asiatic cotton is paleopolyploid.