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- Author or Editor: G. S. Zhang x
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In this work, 9 novel LMW-GS genes (6 LMW-m and 3 LMW-i type) from 4 diploid and 1 tetraploid Aegilops species were amplified and cloned by allelic-specific PCR. Analysis of the deduced amino acid sequences showed that 7 and 2 LMW-GS had 9 and 7 cysteines, respectively. Four LMW-m type subunits genes had an extra cysteine at the C-terminal III, which could form intermolecular disulphide bonds to extend the chains, and therefore would facilitate to form larger gluten polymers. This suggested that these genes are expected to be used as candidate genes for wheat quality improvement. The correlation between specific N-terminal sequences and a decapeptide deletion in the C-terminal II in LMW-GS encoded by D genome was found. Particularly, if LMW-GS possessed a METRCIPG-N-terminal beginning sequences and a decapeptide (LGQCSFQQPQ) deletion in the C-terminal II, they could be encoded by D genome.
Among 20 awnless Tibetan wheat cultivars analyzed by SDS-PAGE, the migration rate of an HMW-GS in XM001584 and XM001593, named 1BX23*. was shown to be slightly faster than 1Bx6. and slower than Bx7. Its nucleotide sequence was isolated based on homology clones. In a phylogenetic tree of 1Bx genes, 1Bx23* was apparently clustered with 1Bx23. Compared with 1Bx23. eight single nucleotide replacements caused four single amino acid replacements in 1Bx23*. The deletion of “G” at base pair 1463 and insertion of “A” at 1509 bps induced a 42-nucleotide frame shift. “GQRQQAGQWQRPGQ” was replaced by “DKGNRQDNGNDRDK”. The new segment cannot be found in other HMW-GSs, and it is very similar to a segment found in collagen. Moreover, an 18-nucleotide deletion made 1Bx23* six amino acids shorter than 1Bx23. The cultivar XM001593 had 28 chromosomes, which signifies that it was tetraploid wheat, and that the new HMW-GS 1Bx23* cannot be used directly for breeding in common wheat.
Gliadin is a main component of gluten proteins that affect functional properties of bread making and contributes to the viscous nature of doughs. In this study, thirteen novel ω-gliadin genes were identified in several Triticum species, which encode the ARH-, ATDand ATN-type proteins. Two novel types of ω-gliadins: ATD- and ATN- have not yet been reported. The lengths of 13 sequences were ranged from 927 to 1269 bp and the deduced mature proteins were varied from 309 to 414 residues. All 13 genes were pseudogenes because of the presence of internal stop codons. The primary structure of these ω-gliadin genes included a signal peptide, a conserved N-terminal domain, a repetitive domain and a conserved C-terminus. In this paper, we first characterize ω-gliadin genes from T. timopheevi ssp. timopheevi and T. timopheevi ssp. araraticum. The ω-gliadin gene variation and the evolutionary relationship of ω-gliadin family genes were also discussed.
In this study, we employed electron microscopy to investigate the cytogenetic and embryologic mechanisms of parthenogenesis induced in the 1BL/1RS male sterile lines of wheat. Analysis of the root tips and acid polyacrylamide gel electrophoresis indicated that all of the male sterile lines and their maintainer lines were 1BL/1RS translocation lines, whereas the restorer lines were non-1BL/1RS translocation lines. Furthermore, the chromosomes of 1BL/1RS wheat lines with T. aestivum cytoplasm and Aegilops cytoplasm (include Ae. kotschyi, Ae. ventricosa, Ae. variabilis) paired abnormally at different rates during meiotic metaphase I (MMI). The translocated segment size of the 1RS chromosome and the specific nuclear–alloplasm interaction impaired the pairing of homologous chromosome in the background of the specific Aegilops cytoplasm at MMI. In addition, the frequency of abnormal chromosomal pairing was directly affected by the frequency of haploid production induced by parthenogenesis. The results of this study provide significant insights into the mechanism of parthenogenesis, which is probably due to the abnormal fertilization of synergid cells in alloplasmic 1BL/1RS wheat.
Studies of the pollen abortion mechanism in thermo-sensitive male sterile lines may provide a strong foundation for breeding hybrid wheat and establishing a theoretical basis for marker-assisted selection. To investigate the cause of pollen abortion in Bainong thermo – sensitive male sterile (BNS) lines, we analyzed the properties of pollen grains, changes in the tapetum and microspores in different anther developmental stages, and the distribution and deposition of nutrient substances in microspores. We found that tapetum degraded in the early uninucleate stage in sterile BNS (S-BNS), which was earlier than that of fertile BNS (F-BNS) tapetum. Large amounts of insoluble polysaccharides, lipids, and proteins were deposited until the trinucleate pollen stage in the nutritive cells in F-BNS. At the binucleate stage, the vacuoles disappeared and pollen inclusion increased gradually. At the trinucleate stage, these nutrients would help pollen grains mature and participate in fertilization normally. Therefore, early degradation of the tapetum, which inhibits normal microspore development, and the limited content of nutrient substances in pollen may be the main factors responsible for male sterility in BNS lines.
Chromosome segment substitution lines (CSSLs) are powerful tools to combine naturally occurring genetic variants with favorable alleles in the same genetic backgrounds of elite cultivars. An elite CSSL Z322-1-10 was identified from advanced backcrosses between a japonica cultivar Nipponbare and an elite indica restorer Xihui 18 by SSR marker-assisted selection (MAS). The Z322-1-10 line carries five substitution segments distributed on chromosomes 1, 2, 5, 6 and 10 with an average length of 4.80 Mb. Spikilets per panicle, 1000-grain weight, grain length in the Z322-1-10 line are significantly higher than those in Nipponbare. Quantitative trait loci (QTLs) were identified and mapped for nine agronomic traits in an F3 population derived from the cross between Nipponbare and Z322-1-10 using the restricted maximum likelihood (REML) method in the HPMIXED procedure of SAS. We detected 13 QTLs whose effect ranging from 2.45% to 44.17% in terms of phenotypic variance explained. Of the 13 loci detected, three are major QTL (qGL1, qGW5-1 and qRLW5-1) and they explain 34.68%, 44.17% and 33.05% of the phenotypic variance. The qGL1 locus controls grain length with a typical Mendelian dominance inheritance of 3:1 ratio for long grain to short grain. The already cloned QTL qGW5-1 is linked with a minor QTL for grain width qGW5-2 (13.01%) in the same substitution segment. Similarly, the previously reported qRLW5-1 is also linked with a minor QTL qRLW5-2. Not only the study is important for fine mapping and cloning of the gene qGL1, but also has a great potential for molecular breeding.
Abstract
Human milk (HM) of healthy, well-nourished, lactating mothers is a unique and ideal source of nutritive factors, like hormones, cytokines, chemokines, growth factors that ensures the proper growth and development of infants. Among the main components of HM, fat is an important energy source and a regulatory factor. The quality of milk fat depends on its fatty acid (FA) composition. Gas chromatography coupled with flame ionisation detection is one of the most common methods for analysis of the FA profile of HM. The aim of this study was to evaluate the FA composition of HM, collected from mothers with different health conditions (normal Body Mass Index (nBMI); overweight and obese) using GC-FID method. The results showed that saturated FAs were present in the highest amount in the HM samples, of which palmitic acid was the main representative. The major monounsaturated FA was oleic acid, while linoleic acid was the most abundant of the polyunsaturated FAs (PUFA). Overweight and obese women have lower levels of PUFA in their breast milk. The data were subjected to principal component and quadratic discriminant analysis (QDA). QDA classified nBMI and overweight and obese mother milk samples with 88.24% accuracy. Significant differences were found between normal and overweight and obese HM samples in case of C10:0 and C18:3 FAs. Higher maternal BMI was associated with a higher n-6/n-3 PUFA ratio.
Abstract
Nattokinase (NK) is effective in the prevention and treatment of cardiovascular disease. Cucumber is rich in nutrients with low sugar content and is safe for consumption. The aim of this study was to construct a therapeutic cucumber that can express NK, which can prevent and alleviate cardiovascular diseases by consumption. Because the Bitter fruit (Bt) gene contributes to bitter taste but has no obvious effect on the growth and development of cucumber, so the NK-producing cucumber was constructed by replacing the Bt gene with NK by using CRISPR/Cas9. The pZHY988-Cas9-sgRNA and pX6-LHA-U6-NK-T-RHA vectors were constructed and transformed into Agrobacterium tumefaciens EHA105, which was transformed into cucumber by floral dip method. The crude extract of NK-producing cucumber had significant thrombolytic activity in vitro. In addition, treatment with the crude extract significantly delayed thrombus tail appearance, and the thrombin time of mice was much longer than that of normal mice. The degrees of coagulation and blood viscosity as well as hemorheological properties improved significantly after crude extract treatment. These findings show that NK-producing cucumber can effectively alleviate thrombosis and improve blood biochemical parameters, providing a new direction for diet therapy against cardiovascular diseases.
Premature termination codons (PTCs) are an important reason for the silence of highmolecular- weight glutenin subunits in Triticum species. Although the Glu-A1y gene is generally silent in common wheat, we here isolated an expressed Glu-A1y gene containing a PTC, named 1Ay8.3, from Triticum monococcum ssp. monococcum (AmAm, 2n = 2x = 14). Despite the presence of a PTC (TAG) at base pair positions 1879–1881 in the C-terminal coding region, this did not obviously affect 1Ay8.3 expression in seeds. This was demonstrated by the fact that when the PTC TAG of 1Ay8.3 was mutated to the CAG codon, the mutant in Escherichia coli bacterial cells expressed the same subunit as in the seeds. However, in E. coli, 1Ay8.3 containing the PTC expressed a truncated protein with faster electrophoretic mobility than that in seeds, suggesting that PTC translation termination suppression probably occurs in vivo (seeds) but not in vitro (E. coli). This may represent one of only a few reports on the PTC termination suppression phenomenon in genes.
