Proteomic and Mutant Analysis of Hydrogenase Maturation Protein Gene hypE in Symbiotic Nitrogen Fixation of Mesorhizobium huakuii.

Hydrogenases catalyze the simple yet important redox reaction between protons and electrons and H2, thus mediating symbiotic interactions. The contribution of hydrogenase to this symbiosis and anti-oxidative damage was investigated using the M. huakuii hypE (encoding hydrogenase maturation protein) mutant. The hypE mutant grew a little faster than its parental 7653R and displayed decreased antioxidative capacity under H2O2-induced oxidative damage. Real-time quantitative PCR showed that hypE gene expression is significantly up-regulated in all the detected stages of nodule development. Although the hypE mutant can form nodules, the symbiotic ability was severely impaired, which led to an abnormal nodulation phenotype coupled to a 47% reduction in nitrogen fixation capacity. This phenotype was linked to the formation of smaller abnormal nodules containing disintegrating and prematurely senescent bacteroids. Proteomics analysis allowed a total of ninety differentially expressed proteins (fold change > 1.5 or <0.67, p < 0.05) to be identified. Of these proteins, 21 are related to stress response and virulence, 21 are involved in transporter activity, and 18 are involved in energy and nitrogen metabolism. Overall, the HypE protein is essential for symbiotic nitrogen fixation, playing independent roles in supplying energy and electrons, in bacterial detoxification, and in the control of bacteroid differentiation and senescence.

Phytoremediation with kenaf (Hibiscus cannabinus L.) for cadmium-contaminated paddy soil in southern China: translocation, uptake, and assessment of cultivars.

Kenaf (Hibiscus cannabinus L.) is suitable for growing in heavy metal-polluted soil for non-food purposes and can be used as a potential crop to remediate heavy metal-contaminated soil. The main objective of this study was to investigate kenaf phytoextraction of cadmium (Cd), including uptake, translocation, and accumulation differences in tissues among kenaf cultivars. A field experiment was conducted in a Cd contaminated paddy field in southern China area with 13 kenaf cultivars in 2015 and 2016. Agronomic performance, Cd concentrations in plant tissues (root, xylem, and phloem), and biomass of different tissues of each cultivar were measured and evaluated. Significant differences in Cd concentrations and accumulation among tissues and cultivars were observed. The phloem had the highest Cd accumulation and transfer capability compared with the roots and xylem. Approximately 35 ~ 65 g of Cd could be taken up by the aerial parts of different kenaf cultivars within every hectare of soil. The percentage of Cd uptake by the phloem ranged from 47 to 61% and by the xylem ranged from 38 to 53%. By evaluating the agronomic traits and Cd bioaccumulation capacity, Fuhong 952, Fuhong 992, and Fuhong R1 were regarded as Cd accumulators for the phytoremediation of Cd-contaminated soil. Our study clearly demonstrated that a significant level of Cd in the soil was taken up through the phytoremediation with kenaf. In addition, harmless utilization of kenaf planting in Cd-contaminated paddy soil was discussed.

Comparative Assessment of Lignan, Tocopherol, Tocotrienol and Carotenoids in 40 Selected Varieties of Flaxseed (Linum usitatissimum L.).

Multiple varieties of flaxseeds have been identified in the world, yet the relationship between these varieties, their agronomic traits, and their seeds' quality remains unclear. This study aimed to determine the level of lignan, vitamins and carotenoids in 40 selected flaxseed varieties, and the relationship between varieties, agronomic traits, and seed quality was investigated. In this study, notably, fiber flax variety No. 225 exhibited the highest lignan content among all tested seeds. Additionally, oil variety No. 167 demonstrated the highest level of α-tocotrienol (α-T3), ß-tocopherol (ß-T), γ-tocotrienol (γ-T3), and ß-carotene (ß-Car.). Conversely, intermediate flax variety No. 16 displayed the highest content of α-tocopherol (α-T), but lowest content of lutein (Lut.), zeaxanthin (Zea.), ß-carotene (ß-Car.), and total carotenoids (Total Car.). Furthermore, a correlation was observed between petal color with the lignan, while a strong correlation has been explored in seed yield, seed type, plant natural height, and fiber content in straw. Nevertheless, further investigation is required to elucidate the internal relationship between varieties with compositions.

Cadmium accumulation, translocation, and assessment of eighteen Linum usitatissimum L. cultivars growing in heavy metal contaminated soil.

Flax (Linum usitatissimum L.) is suitable for growing in heavy metal polluted soil for non-food purposes and can be used as potential crops for cleaning the soil from heavy metals. The main objective of this study was to investigate flax/linseed phytoextraction of cadmium (Cd), including uptake, translocation, and accumulation differences in organs among flax/linseed cultivars. A field experiment was carried out in Cd contaminated field of Chinese southern area with nine flax, one dual-purpose, and eight linseed cultivars. Cd concentrations in plant organs (root, xylem, phloem, and capsule) and biomass of different organs of each cultivar were measured and evaluated. Significant differences in Cd concentration and accumulation among organs were observed. The most Cd was accumulated by phloem, which was 2 ∼ 4 times more than other organs, followed by roots, xylems and capsules played a comparably smaller role. The uptake of Cd by flax/linseed from per hectare was calculated and had highly significant positive correlation with biomass. Phloem accumulated 45 ∼ 55% of total Cd from the soil by the plant. Among testing 18 flax/linseed cultivars, one flax cultivar (Zhongya 1), and two linseed cultivars (Y2I329 and Y2I330), which extracted more than 60 g ha-1, can be considered as Cd accumulators for phytoremediation of Cd contaminated soil.

Digital gene expression profiling of flax (Linum usitatissimum L.) stem peel identifies genes enriched in fiber-bearing phloem tissue.

To better understand the molecular mechanisms and gene expression characteristics associated with development of bast fiber cell within flax stem phloem, the gene expression profiling of flax stem peels and leaves were screened, using Illumina's Digital Gene Expression (DGE) analysis. Four DGE libraries (2 for stem peel and 2 for leaf), ranging from 6.7 to 9.2 million clean reads were obtained, which produced 7.0 million and 6.8 million mapped reads for flax stem peel and leave, respectively. By differential gene expression analysis, a total of 975 genes, of which 708 (73%) genes have protein-coding annotation, were identified as phloem enriched genes putatively involved in the processes of polysaccharide and cell wall metabolism. Differential expression genes (DEGs) was validated using quantitative RT-PCR, the expression pattern of all nine genes determined by qRT-PCR fitted in well with that obtained by sequencing analysis. Cluster and Gene Ontology (GO) analysis revealed that a large number of genes related to metabolic process, catalytic activity and binding category were expressed predominantly in the stem peels. The Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of the phloem enriched genes suggested approximately 111 biological pathways. The large number of genes and pathways produced from DGE sequencing will expand our understanding of the complex molecular and cellular events in flax bast fiber development and provide a foundation for future studies on fiber development in other bast fiber crops.

Analysis of 2,297 expressed sequence tags (ESTs) from a cDNA library of flax (Linum ustitatissimum L.) bark tissue.

Bast fibre crops are the second most important natural fibre crops following cotton. Of these, flax (Linum ustitatissimum L.) is the most widely planted in the world, with its fibre used for high quality linen textile. A cDNA library of flax bark tissues was constructed with the purpose of identifying genes involved in the Bast fibre development. A total of 2,297 unigene sequences were obtained from 3,200 randomly selected clones of the cDNA library. These sequences were grouped into 155 clusters and 2,142 singletons, which have been submitted to the GenBank databases. By putative functional annotation, 23.3% of these sequences were similar to known proteins in GenBank, 44.0% of these sequences were similar to unknown proteins, and 32.7% of these sequences showed no significant similarity to any other protein sequences in existing databases. Classified by the Gene Ontology, 24.8, 23.1 and 14.3% were assigned to molecular function, biological process, and cellular component GO terms, respectively. By further bioinformatics approaches, about 110 ESTs matched cell wall related genes in the MAIZEWALL database, representing 16 functional categories of all 19 categories, of which, the most abundant category was protein synthesis. Based on the PlantTFDB database, 39 of the 64 transcription factor families in the Arabidopsis thaliana genome were identified as being involved in flax cell wall formation. The sequences and bioinformatics analysis data generated in this paper will be useful for gene expression, cloning and genetic engineering studies to characterize bast fibre development and improve the properties of the bast fibres.

Development and characterization of polymorphic microsatellite markers in Linum usitatissimum.

Thirty-five microsatellite loci were isolated and characterized in Linum usitatissimum using enriched genomic libraries. These loci were screened in eight cultivars from different countries and regions and were found to be polymorphic, with the number of alleles per locus ranging from two to six, and observed and expected heterozygosities ranging from 0.125 to 0.375 (mean 0.013) and from 0.233 to 0.842 (mean 0.601), respectively. These polymorphic new microsatellite loci will be useful for genetic linkage map construction, germplasm classification and identification, gene identification and quantitative trait loci mapping, and marker-assisted selection in breeding in L. usitatissimum.

[Construction of a cDNA library of Alternaria sp. strain JH505 using Gateway technology].

A cDNA library was constructed based on the Gateway system that is a method to construct a high-quality cDNA library without the use of restriction enzyme for cloning. This is the first report that the Gateway system is used to construct a cDNA library for Alternaria sp.. The entry cDNA library was constructed in this report has a high titer of 1 x 10(7) cfu/mL and contain a total clones of 9 x 10(7) cfu, with an average inserts size of about 1510bp. In order to screen for a gene encoding the plant activator protein from library using an antiserum, the entry cDNA library was transferred into Gateway destination vector to create an expression library. The expression library show a high titer of 1.58 x 10(6) cfu/mL and contains a total clones of 6.32 x 10(6) cfu, with an average inserts size of 1680bp.