Effect of selenium-containing biocomposites based on <Emphasis Type="Italic">Ganoderma</Emphasis> mushroom isolates grown in the presence of oxopropyl-4-hydroxycoumarins,on bacterial phytopathogens

Effect of selenium-containing biocomposites obtained from submerged cultures of macrobasidiomycetes Ganoderma applantum, G. cattienensis, G. colossus G. lucidum, G. neojaponicum, and G. valesiacum, on plant pathogenic bacteria Clavibacter michiganensis ssp. sepedonicus (Cms), Micrococcus luteus, Pectobacterium atrosepticum, Pectobacterium carotovorum subsp. carotovorum, Pseudomonas fluorescens, Pseudomonas viridiflava, and Xanthomonas campestris was studied. Oxopropyl-4-hydroxychromenones were used as components of the fungal nutrient media. The bacteriostatic and bactericidal activity of the Se-containing and Se-free substances of fungal origin against plant pathogenic bacteria was determined using colony-forming units count, the agar well diffusion method, and by turbidity measurements of bacterial suspensions. The composites produced from the extracellular metabolites of G. cattienensis SIE1302 with 4-hydroxy-3-(3-oxo-1,3-diphenyl propyl)-chromen-2-one (S(45)), and of G. lucidum SIE1303 with 4-hydroxy-3-(3-oxo-1-(3-nitrophenyl)-3-phenylpropyl)-chromen-2-one (S(NO₂)) possessed the most pronounced antibacterial action against Cms. The composites produced from the isolates of G. valesiacum 120702 with S(NO₂) showed the maximal antibacterial activity against Xanthomonas campestris B-610. High antimicrobial effect of G. lucidum 1315 with S(NO₂) against Xanthomonas campestris B-610 and of G. colossus SIE1301 against Pseudomonas fluorescens EL-2.1 was revealed. The pioneering information on the biological activity of coumarin series compounds in their application for producing the substances of fungal origin was obtained.     

Molecular characterisation of two novel starch granule proteins 1 in wild and cultivated diploid A genome wheat species

Starch synthase IIa, also known as starch granule protein 1 (SGP-1), plays a key role in amylopectin biosynthesis. The absence of SGP-1 in cereal grains is correlated to dramatic changes in the grains’ starch content, structure, and composition. An extensive investigation of starch granule proteins in this study revealed a polymorphism in the electrophoretic mobility of SGP-1 between two species of wheat, Triticum urartu and T. monococcum; this protein was, however, conserved among all other Triticum species that share the A genome inherited from their progenitor T. urartu. Two different electrophoretic profiles were identified: SGP-A1 proteins of T. urartu accessions had a SDS–PAGE mobility similar to those of tetraploid and hexaploid wheat species; conversely, SGP-A1 proteins of T. monococcum ssp. monococcum and ssp. boeoticum accessions showed a different electrophoretic mobility. The entire coding region of the two genes was isolated and sequenced in an attempt to explain the polymorphism identified. Several single nucleotide polymorphisms (SNPs) responsible for amino acid changes were identified, but no indel polymorphism was observed to explain the difference in electrophoretic mobility. Amylose content did not differ significantly among T. urartu, T. monococcum ssp. boeoticum and T. monococcum ssp. monococcum, except in one accession of the ssp. boeoticum. Conversely, several interspecific differences were observed in viscosity properties (investigated as viscosity profiles using a rapid visco analyzer—RVA profiles) of these cereal grains. T. monococcum ssp. boeoticum accessions had the lowest RVA profiles, T. urartu accessions had an intermediate RVA profile, whereas T. monococcum ssp. monococcum showed the highest RVA profile. These differences could be associated with the numerous amino acid and structural changes evident among the SGP-1 proteins.     

Protein characters and relationship betweenPhaseolus vulgaris ssp.Aborigineus Burk. and related taxons of the genusPhaseolus

Both quantitative and qualitative immunochemical methods were used for studying the mutual relationships of several spocies and the subspecies of the genusPhaseolus: Ph. vulgaris L. ssp.vulgaris, Ph. vulgaris L. ssp.aborigineus Burk.,Ph. coccineus L.,Ph. acutifolius A. Gray,Ph. lunatus L. (American endemites) andPh. aureus L. (a typical Asian bean). Protein characters of cotyledons (i.e., ?storage” proteins) of the above species were compared with the aid of antisera prepared against seed (cotyledon) proteins ofPh. vulgaris L. ssp.vulgaris, cv. Veltruská Saxa, using

1. (a)
   the whole complex of cotyledon protein,     
   
   

Protein markers for identification of different species and varieties of cotton

Reference electrophoretic spectra that allow compiling electrophoretic formulas of certain cotton species and varieties were obtained on the basis of analysis of the electrophoretic spectrum of water-soluble and barely soluble proteins of seeds of diploid cotton species of genomic group A (Gossypium arboreum var. indicum, G. arboreum ssp. obtusifolum, G. herbaceum ssp. africanum, and G. herbaceum Harga), group C (G. australe, G. bickii, G. nelsone, and G. sturtianum), group D (G. davidsonii, G. harknessii, G. klotzschianum, G. raimondii, G. thurberi, and G. trilobum), and amphidiploid species of group AD (G. mustelinum, G. hirsutum ssp. palmeri, G. tricuspidatum Bagota, G. tricuspidatum Mari Galanta, G. barbadense L., and G. hirsutum L.)     

The evolution of cultivated potatoes

In the first stages of potato evolution in the northern Andes, diploid cultivated species of theSolanum stenotomum complex were selected, in all probability, from wild progenitors in theS. brevicaule complex. TetraploidSolanum tuberosum ssp.andigena arose by fusion of unreduced gametes of a parent in theS. stenotomum complex with those of an unidentified wild species having actinomorphic calyces. Unreduced male gametes of several diploid species fertilized eggs of ssp. andigena leading to extensive introgression.Solanum tuberosum ssp.tuberosum probably originated from a cross between ssp.andigena as staminate parent and an unidentified wild species which contributed cytoplasmic sterility factors encoded in mitochondria and/or plastids having a distinctive type of DNA. Derivatives of this hybridization, which may have occurred in northwestern Argentina, evolved to ssp. tuberosum in southern Chile and southern Argentina. In the 1570’s ssp.andigena was imported to Europe and spread from there to become a major crop with worldwide distribution. In the 1840's it was essentially eliminated by late blight,Phytophthora infestans.Solanum tuberosum ssp.tuberosum was introduced from Chile into North America and Europe in the late 1800's, and in turn achieved a worldwide distribution, filling the vacated agricultural niche of ssp.andigena. The differences between ssp.andigena and ssp.tuberosum in South America are sufficient that the two could reasonably be considered to be separate species. Since the 1960’s the two taxa have been hybridized often in breeding programs. Neotuberosum, a northern-adapted strain of ssp. andigena, has been selected to mimic ssp.tuberosum. Substitution back-cross products have been produced that have the chromosomal genes of ssp.tuberosum combined with cytoplasmic factors of Andean species. These breeding activities are blurring the distinctions between the two subspecies throughout much of the world, though they remain distinct in their native areas in South America.     

Seed protein fraction electrophoresis in peanut (<Emphasis Type="Italic">Arachis hypogaea</Emphasis> L.) accessions and wild species

Total seed storage proteins were studied in 50 accessions of A. hypogaea (11 A. hypogaea ssp. hypogaea var hypogaea, 13 A. hypogaea ssp. hypogaea var hirsuta, 11 A. hypogaea ssp. fastigiata var fastigiata and 15 A. hypogaea ssp. fastigiata var. vulgaris accessions) in SDS PAGE. These accessions were also analysed for albumin and globulin seed protein fractions. Among the six seed protein markers presently used, it was found that globulin fraction showed maximum diversity (77.2%) in A. hypogaea accessions followed by albumin (52.3%), denatured total soluble protein fraction in embryo (33.3%) and cotyledon (28.5%). The cluster analysis based on combined data of cotyledons, embryos, albumins and globulins seed protein fractions demarcated the accessions of two subspecies hypogaea and fastigiata into two separate clusters supported by 51% bootstrap value, with few exceptions, suggesting the genotypes to be moderately diverse. Native and denatured total soluble seed storage proteins were also electrophoretically analysed in 27 wild Arachis species belonging to six sections of the genus. Cluster analysis using different methods were performed for different seed proteins data alone and also in combination. Section Caulorrhizae (C genome) and Triseminatae (T genome) formed one, distantly related group to A. hypogaea and other section Arachis species in the dendrogram based on denatured seed storage proteins data. The present analysis has maintained that the section Arachis species belong to primary and secondary genepools and, sections Procumbenetes and Erectoides belong to tertiary gene pools.     

Generation of Chinese cabbage resistant to bacterial soft rot by heterologous expression of <Emphasis Type="Italic">Arabidopsis WRKY75</Emphasis>

Soft rot caused by Pectobacterium carotovorum subsp. carotovorum (Pcc) is a serious disease in Chinese cabbage (Brassica rapa L. subsp. pekinensis). To reduce the severity of soft rot symptoms in Chinese cabbage, Arabidopsis AtWRKY75 was introduced into Chinese cabbage by Agrobacterium-mediated transformation, which was previously reported to reduce susceptibility to Pcc infection in Arabidopsis. Three independent Chinese cabbage transgenic lines carrying AtWRKY75 were obtained. The growth phenotypes of AtWRKY75 overexpression (OE) lines were normal. Bacterial soft rot symptoms and Pcc growth were reduced in AtWRKY75-OE Chinese cabbage lines compared with WT plants. In contrast, overexpression of AtWRKY75 had no effect on infection with a hemibiotrophic pathogen, Xanthomonas campestris pv. campestris (Xcc) causing black rot disease. These results are consistent with those observed in the transgenic Arabidopsis. We found that AtWRKY75 activated a subset of Chinese cabbage genes related to defense against Pcc infection, such as Meri15B, BrPR4, and BrPDF1.2 (but not BrPGIP2). Moreover, overexpression of AtWRKY75 caused H₂O₂ production and activation of H₂O₂ scavenge enzyme genes, suggesting that H₂O₂ played a role in AtWRKY75-mediated resistance to Pcc. Together, these results demonstrated that AtWRKY75 decreased the severity of Pcc-caused bacterial soft rot and activated a subset of Pcc infection defense-related genes in Chinese cabbage similar to in Arabidopsis. It is suggested that AtWRKY75 is a candidate gene for use in crop improvement, because it results in reduced severity of disease symptoms without concurrent growth abnormalities.     

Identification of the di-<Emphasis Type="Italic">n</Emphasis>-butyl phthalate-biodegrading strains and the biodegradation pathway in strain LMB-1

DNA isolated from a greenhouse soil (Nanjing, Jiangsu Province, China) was suitable for PCR amplification of gene segment coding for the 16S rRNA. Diverse PCR products were characterized by cloning and sequencing, and analysis of bacterial colonies showed the presence over 26 phyla. The most bacteria belonged to Proteobacteria, Actinobacteria, Gemmatimonadetes, Acidobacteria and Planctomycetes. Furthermore, after the enrichment procedure of DBP-degrading microorganisms, 4 strains were isolated from the soil sample with di-n-butyl phthalate (DBP) biodegradability, and they were identified to be Rhizobium sp., Streptomyces sp., Pseudomonas sp. and Acinetobacter sp. Analysis of the degradation products by LC-MS led to identification of metabolites of DBP in strain LMB-1 (identified as Rhizobium sp.) which suggests that DBP was degraded through β-oxidation, demethylation, de-esterification and cleavage of aromatic ring.     

Antimicrobial Activity of Geometric Isomers of Etherolenic Acid—the Products of Plant Lipoxygenase Cascade

Data on the influence of the double bond geometry on the antimicrobial properties of different isomers of etherolenic acid against phytopathogenic bacteria are presented. (ω5Z)-Etherolenic acid possesses bactericidal properties against Xanthomonas campestris ssp. vesicatoria, Pseudomonas syringae ssp. tomato, Pectobacterium atrosepticum SCRI1043; the etherolenic and (11Z)-etherolenic acids possess only bacteriostatic properties.     

Development of cost-effective single nucleotide polymorphism marker assays for genetic diversity analysis in <Emphasis Type="Italic">Brassica rapa</Emphasis>

Competitive allele-specific PCR (KASPar) assay is a user-friendly system that provides flexibility in the numbers of single nucleotide polymorphisms (SNPs) and genotypes. Based on Illumina-GA-IIx genomic data from 10 genotypes with a broad genetic background, 3183 SNPs were selected for KASPar assays development, and 568 were finally converted and selected for Brassica rapa germplasm characterization (17.8%) on the basis of reproducibility, missing data rate, and uniform genetic distribution. High levels of polymorphism of these markers across 231 B. rapa genotypes were verified, illustrating by high polymorphic information content (averaged 0.34), minor allele frequency (0.37), genetic diversity (0.45), and the low observed heterozygosity (0.10). Based on the SNP dataset, structure and principal coordinates analysis, and neighbor-joining phylogenetic methods were used to examine the population structure and gave highly consistent results. The 231 accessions were divided into the four primary subspecies, representing 99 accessions from B. rapa ssp. pekinensis, 85 from B. rapa ssp. chinensis, 30 from B. rapa ssp. rapifera, and 17 from B. rapa ssp. oleifera and were further subdivided into 12 lower-order clusters according to different morphotypes. The genetic variability and pairwise fixation index analysis revealed that the ssp. pekinensis accessions possess the most extensive genetic variation among the four subspecies. The KASPar system is highly useful for validating SNPs and will be valuable for genetics research and breeding applications in B. rapa.     

Diversity array technology (DArT) 56K analysis,confirmed by SNP markers,distinguishes one сrested wheatgrass <Emphasis Type="Italic">Agropyron</Emphasis> species from two others found in Kazakhstan

Molecular genetic polymorphism in three species and four subspecies of crested wheatgrass, Agropyron, was studied using 56K diversity array technology (DArT), and the results confirmed with four selected SNP Amplifluor markers. In total, 82 accessions from three species—A. desertorum, A. fragile, and two subspecies of A. cristatum (ssp. cristatum and ssp. pectinatum)—were collected from various regions of Kazakhstan or ordered from Genebank in Russia, for morphological taxonomy and molecular phylogenetic analyses. In the DArT clone analysis, two Agropyron species with narrow linear spikes, A. fragile and A. desertorum, were found to be genetically similar and fell within a single clade (A). Both species share similar eco-geographical origins. All samples of A. cristatum including the two subspecies, ssp. pectinatum and ssp. cristatum, which have short broad spikes, were interspersed within two other clades, B and C, more genetically distanced from the other species. Four SNP Amplifluor markers developed for genetic fragments on different chromosomes confirmed the distinction between the studied species. These results, derived from multiple molecular markers, suggest that the morphological taxonomy of these Agropyron species should be re-considered carefully in the future.     

Heteroplasmy due to chloroplast paternal leakage: another insight into <Emphasis Type="Italic">Phragmites</Emphasis> haplotypic diversity in North America

Chloroplasts contain several copies of their DNA, and intra-individual haplotypic variation (heteroplasmy) is common in plants, but unexplored in the cosmopolitan genus Phragmites. The aims of this study were to assess if heteroplasmy due to paternal leakage of the chloroplast occurs in Phragmites and which new insights into the evolutionary history of Phragmites australis in North America can be identified from the heteroplasmic variation. Eight non-native P. australis haplotypes occur in North America and can interbreed with P. australis ssp. americanus and P. australis var. berlandieri, creating opportunities for biparental inheritance of distinctive haplotypes. The polymorphism in the trnT-trnL sequence length revealed seventeen cases of heteroplasmy worldwide, in contact zones of distantly related haplotypes and in known hybrid populations, nine of which occurred in North America. In America, the cloned sequences, combined with nuclear markers, identified recombined haplotypes between native P. australis ssp. americanus and invasive P. australis haplotype M, and between the species P. mauritianus and P. australis, due to chloroplast paternal leakage. The occurrence of heteroplasmy and recombined haplotypes suggest a local origin for some of the rare non-native haplotypes occurring in North America, and plastid leakage events in the evolutionary histories of P. australis ssp. americanus and P. australis var. berlandieri.     

Heat stress differentially modifies ethylene biosynthesis and signaling in pea floral and fruit tissues

Key message

We identified and cloned the two precursors of miR158 and its target gene in Brassica campestris ssp. chinensis, which both had high relative expression in the inflorescences. Further study revealed that over-expression of miR158 caused reduced pollen varbility, which was caused by the degradation of pollen contents from the binucleate microspore stage. These results first suggest the role of miR158 in pollen development of Brassica campestris ssp. chinensis.

Abstract

MicroRNAs (miRNAs) play crucial roles in many important growth and development processes both in plants and animals by regulating the expression of their target genes via mRNA cleavage or translational repression. In this study, miR158, a Brassicaceae specific miRNA, was functionally characterized with regard to its role in pollen development of non-heading Chinese cabbage (Brassica campestris ssp. chinensis). Two family members of miR158 in B. campestris, namely bra-miR158a1 and bra-miR158a2, and their target gene bra027656, which encodes a pentatricopeptide repeat (PPR) containing protein, were identified. Then, qRT-PCR analysis and GUS-reporter system revealed that both bra-miR158 and its target gene had relatively high expression levels in the inflorescences. Further study revealed that over-expression of miR158 caused reduced pollen varbility and pollen germination ratio, and the degradation of pollen contents from the binucleate microspore stage was also found in those deformed pollen grains, which led to pollen shrinking and collapse in later pollen development stage. These results first shed light on the importance of miR158 in pollen development of Brassica campestris ssp. chinensis.

     

Stable expression of exogenous imported <Emphasis Type="Italic">sporamin</Emphasis> in transgenic Chinese cabbage enhances resistance against insects

Cultivating insect pest-resistant varieties is one of the most effective ways to prevent or mitigate pest infestation in Chinese cabbage (Brassica campestris ssp. chinensis). Via the agrobacterium tumefaciens-mediated transformation method, this study introduced the protease inhibitor encoding gene sporamin into two widely cultured cultivars ‘Youdonger’ and ‘Shanghaiqing’, of the common variety of Chinese cabbages (B. campestriss ssp. chinensis var. communis), getting transgenic plants with high sporamin expression. In vitro insect bioassays indicated that, compared with the wild type plants, the transgenic plants exhibited improved resistance to diamondback moth (Plutella xylostella L.) The analysis of inheritance pattern of exogenous sporamin in the progenies of single copy insertion transgenic lines demonstrated that sporamin could be inherited and expressed stably in transgenic progenies. Field survey of the insect resistance under the normal culture condition confirmed the enhanced resistance in transgenic progenies to diamondback moth. Our results strongly suggest that sporamin is an efficient candidate gene for insect-resistant genetic engineering in Chinese cabbage.     

<Emphasis Type="Italic">ZmCCD7/ZpCCD7</Emphasis> encodes a carotenoid cleavage dioxygenase mediating shoot branching

Main conclusion

ZmCCD7/ZpCCD7 encodes a carotenoid cleavage dioxygenase that may mediate strigolactone biosynthesis highly responsive to phosphorus deficiency and undergoes negative selection over domestication from Zea ssp. parviglumis to Zea mays.Carotenoid cleavage dioxygenase 7 (CCD7) functions to suppress shoot branching by controlling strigolactone biosynthesis. However, little is known about CCD7 and its functions in maize and its ancestor (Zea ssp. parviglumis) with numerous shoot branches. We found that ZmCCD7 and ZpCCD7 had the same coding sequence, indicating negative selection of the CCD7 gene over domestication from Zea ssp. parviglumis to Zea mays. CCD7 expression was highly responsive to phosphorus deficiency in both species, especially in the meristematic zone and the pericycle of the elongation zone of maize roots. Notably, the crown root had the strongest ZmCCD7 expression in the meristematic zone under phosphorus limitation. Transient expression of GFP tagged ZmCCD7/ZpCCD7 in maize protoplasts indicated their localization in the plastid. Further, ZmCCD7/ZpCCD7 efficiently catalyzed metabolism of six different linear and cyclic carotenoids in E. coli, and generated β-ionone by cleaving β-carotene at the 9,10 (9′,10′) position. Together with suppression of shoot branching in the max3 mutant by transformation of ZmCCD7/ZpCCD7, our work suggested that ZmCCD7/ZpCCD7 encodes a carotenoid cleavage dioxygenase mediating strigolactone biosynthesis in maize and its ancestor.

     

Tandem duplications of receptor-like kinase genes reshaped the homologous chromosome 1 regions in <Emphasis Type="Italic">Oryza sativa</Emphasis> ssp. <Emphasis Type="Italic">indica</Emphasis> and <Emphasis Type="Italic">japonica</Emphasis> and their possible ancestral genomes

Plant receptor-like kinase (Rlk) genes form a large family, each encoding a protein with a signal motif, a single transmembrane region, and a cytoplasmic kinase domain. Various gene duplications have contributed to the establishment and expansion of the family. Here, we characterized the formation and evolution of the Rlk gene family in cultivated rice and their possible progenitors. Using wheat Rlk gene sequences, we identified orthologs from the genomes of domesticated rice subspecies Oryza sativa ssp. japonica and ssp. indica and their putative progenitors O. glaberrima and O. rufipogon. The four chromosome 1 orthologous regions ranged from 103 to 281 kb comprising 181 syntenic blocks with 75 to 100% sequence identity. These regions contained 11–19 Triticum aestivum kinases (Taks) and 10–15 Lr10 receptor-like kinases (Lrks) organized in clusters and 3–12 transposable elements (TEs). Dot plot analyses showed that the 4 regions had 21–37 conserved catalytic domains, mainly in protein kinases (PKs) and tyrosine kinases (TyrKs) in coupling state. Over 50% of the sequences of glaberrima/rufipogon and japonica/indica pairs were colinear, while japonica/indica displayed a marked sequence expansion with duplicated genes and TEs. A total of 2312 single nucleotide polymorphisms (SNPs) and insertion-deletions (INDELs) were identified between japonica and indica. Duplication of the Rlk genes in O. glaberrima and O. rufipogon occurred after the grass species radiation and before the divergence of O. rufipogon from O. glaberrima; the orthologous Rlk genes from O. japonica and O. indica duplicated after O. sativa separated from O. rufipogon; paralogs, obtained through extensive duplication, happened after the separation of rice from maize. Tandem duplication was the major factor contributing to the gene copy number variation and genome size expansion.     

Differential expression of two <Emphasis Type="Italic">CONSTANS-LIKE1</Emphasis> genes in potato

Although the CONSTANS gene and its CONSTANS-LIKE1 (COL1) orthologs are known to control the photoperiod-dependent floral transition in many plant species, the role of these genes in Solanum development has not been sufficiently elucidated. Previously we characterized two forms of CONSTANS-LIKE1 genes, sCOL1 and lCOL1, in potato (Solanum tuberosum ssp. tuberosum). To prove that these genes were functional, we followed their expression in potato cv. Early Rose with the real-time PCR technique. Both sCOL1 and lCOL1 displayed characteristic day-night patterns of expression under long-day and short-day conditions. The profiles and amplitudes of expression dramatically differed in two genes, with the maximum sCOL1 expression exceeding that of lCOL1 by an order of magnitude.