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Leaf senescence is a notably important trait that limits the yield and biomass accumulation of agronomic crops. Therefore, determining the chromosomal position of the expression sequence tags (ESTs) that are associated with leaf senescence is notably interesting in the manipulation of leaf senescence for crop improvement. A total of 32 ESTs that were previously identified during the delaying leaf senescence stage in the stay-green wheat cultivar CN17 were mapped to 42 chromosomes, a chloroplast, a mitochondrion, and a ribosome using in silico mapping. Then, we developed 19 pairs of primers based on these sequences and used them to determine the polymorphisms between the stay-green cultivars (CN12, CN17, and CN18) and the control cultivar MY11. Among the 19 pairs of primers, 5 pairs produced polymorphisms between the stay-green cultivar and the non-stay-green control. Further studies of Chinese Spring nullisomic-tetrasomics show that JK738991 is mapped to 3B, JK738983 is mapped to 5D, and JK738989 is mapped to 2A, 4A, and 3D. The other two ESTs, JK738994 and JK739003, were not assigned to a chromosome using the Chinese Spring nullisomic-tetrasomics, which indicates that these ESTs may be derived from rye DNA in the wide cross. In particular, the ESTs that produce polymorphisms are notably useful in identifying the stay-green cultivar using molecular marker-assisted selection. The results also suggest that the in silico mapping data, even from a comparison genomic analysis based on the homogeneous comparison, are useful at some points, but the data were not always reliable, which requires further investigation using experimental methods.

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Journal of Thermal Analysis and Calorimetry
Authors:
F. Xu
,
L. Sun
,
P. Chen
,
Y. Qi
,
J. Zhang
,
J. Zhao
,
Y. Liu
,
L. Zhang
,
Zhong Cao
,
D. Yang
,
J. Zeng
, and
Y. Du

Abstract  

The heat capacities of LiNH2 and Li2MgN2H2 were measured by a modulated differential scanning calorimetry (MDSC) over the temperature range from 223 to 473 K for the first time. The value of heat capacity of LiNH2 is bigger than that of Li2MgN2H2 from 223 to 473 K. The thermodynamic parameters such as enthalpy (HH 298.15) and entropy (SS 298.15) versus 298.15 K were calculated based on the above heat capacities. The thermal stabilities of them were investigated by thermogravimetric analysis (TG) at a heating rate of 10 K min−1 with Ar gas flow rate of 30 mL min−1 from room temperature to 1,080 K. TG curves showed that the thermal decomposition of them occurred in two stages. The order of thermal stability of them is: Li2MgN2H2 > LiNH2. The results indicate that addition of Mg increases the thermal stability of Li–N–H system and decrease the value of heat capacities of Li–N–H system.

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Summary

A selective and sensitive liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS) method was developed and validated for analysis of xanthotoxol (1), xanthotoxin (2), isoimpinellin (3), bergapten (4), oxypeucedanin (5), imperatorin (6), cnidilin (7), and isoimperatorin (8) in rat bile and urine using pimpinellin as an internal standard (IS). An Agilent 1200 liquid chromatography system (Agilent Technologies, USA) equipped with a quaternary pump, an autosampler, and a column compartment was used for all analyses. Chromatographic separations were performed on a Sapphire C18 column (150 mm × 4.6 mm, 5 μm), and the column temperature was maintained at 30°C; the sample injection volume was 10 μL. The specificity, linearity, accuracy, precision, recovery, matrix effect, and several stabilities were validated for all analytes in the rat bile and urine samples. The method was successfully applied in monitoring the concentrations of eight coumarins in rat bile and urine after a single oral administration of Radix Angelicae Dahuricae extract with a dosage of 8.0 mL/kg. In the bile samples, the eight coumarins excreted completely in twenty-four hours. The average percentages of coumarins (1–8) excreted were 0.045%, 0.019%, 0.177%, 0.105%, 0.337%, 0.023%, 0.024%, 0.021%. In the urine samples, the eight coumarins excreted completely in seventy-two hours. The average percentages of coumarins (1–8) excreted were 1.78%, 0.095%, 0.130%, 0.292%, 0.082%, 0.008%, 0.005%, 0.004%. The method is robust and specific and it can successfully complete the requirements of the excretion study of the eight coumarins in Radix Angelicae Dahuricae.

Open access
Cereal Research Communications
Authors:
X. Zhang
,
Y. Chen
,
Y. Wei
,
W. Lu
,
H. Liao
,
Y. Liu
,
X. Yang
,
X. Li
,
L. Yang
,
L. Li
, and
R. Li

Partial abortion of gametes possessing S-5 j in S-5 i / S-5 j genotype at locus S-5 is responsible for hybrid sterility between indica and japonica subspecies in rice ( Oryza sativa L.), while a single wide compatibility (WC) allele S-5 n can restore normal hybrid fertility between the two groups. In this study, Pei’ai 64S, one of the most popular WC line widely used for subspecific hybrid rice breeding program in South China was studied for location of its S-5 locus. Twenty SSR (Simple Sequence Repeat) markers derived from Cornell SSR linkage map and 9 developed using sequences from GenBank database were employed to perform bulked segregant analysis of the mapping population derived from a three-way cross (Pei’ai 64S/T8//Akihikari) to tag fine location of the hybrid sterility locus, S-5 . This S-5 locus was mapped on chromosome 6 approximately 0.2 cM from GXR6 and RM276 SSR markers. This tight linkage of the markers and the S-5 locus would be very useful for efficient marker-assisted selection for WC varieties and for map-based cloning of the gene.

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Red coleoptile is an easily observed trait in Triticum aestivum and can provide some protection against stress. Here, TaMYB-A1 or TuMYB-A1, homologous to TaMYB-D1, which controls red coleoptile formation in the common wheat cultivar ‘Gy115’, was isolated from eight T. aestivum and 34 T. urartu cultivars. The genome sequence of TaMYB-A1 was 867 bp with an intron of 93 bp, which was similar to the MYBs regulating anthocyanin biosynthesis in T. aestivum but different from other MYB transcription factors regulating anthocyanin biosynthesis. TaMYB-A1 had an integrated DNA-binding domain of 102 amino acids and a transcriptional domain of 42 amino acids, which was responsible for regulating anthocyanin biosynthesis. TaMYB-A1 was assigned to the same branch as the MYBs regulating anthocyanin biosynthesis in a phylogenetic tree. A transient expression analysis showed that TaMYB-A1 induced ‘Opata’ coleoptile cells to synthesize anthocyanin with the help of ZmR. A non-functional allele of TaMYB-a1 existed in common wheat cultivars containing rc-a1. One single nucleotide was deleted 715 bp after the start codon in TaMYB-a1 compared with TaMYB-A1. The deletion caused a frame shift mutation, destroyed the DNA transcription activator domain, and resulted in TaMYB-a1 losing its ability to regulate anthocyanin biosynthesis in ‘Opata’ coleoptile cells. Those cultivars with functional TaMYB-A1 or TuMYB-A1 have red coleoptiles. The isolation of TaMYB-A1 should aid in understanding the molecular mechanisms of coleoptile traits in T. aestivum.

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An efficient and sensitive analytical method based on precolumn derivatization and gas chromatography—mass spectrometry—selected ion monitoring (GC—MS—SIM) was proposed and validated for analysis of two cembrenediols (CBDs) which are α-cembrenediol and β-cembrenediol in tobacco samples. CBDs in tobacco samples were extracted by sonication with 50 mL dichloromethane for 10 min before derivatized with 2:3 (v/v) bis(trimethylsilyl)trifluoroacetamide (BSTFA)—pyridine at 20 °C for 100 min. CBDs’ level in tobacco samples was analyzed by GC—MS—SIM and quantified by the internal standard method. The linear range for α-CBD and β-CBD was 13.6–554.6 μg mL−1 and 4.11–162.6 μg mL−1, and the correlation coefficients of both were 0.9998. The limit of detection (LOD) and limit of quantification (LOQ) of α-cembrenediol and β-cembrenediol were 0.40 μg g−1 and 1.34 μg g−1, and 0.27 μg g−1 and 0.90 μg g−1, respectively. Average recoveries of α-CBD and β-CBD were 94.4–99.9% and 91.9–98.2% while the relative standard deviations (RSDs, n = 5) were ranged from 2.67 to 5.6% and 2.04 to 4.22%, respectively. This proposed analytical method has been successfully applied to analyze CBDs in tobacco samples.

Open access
Cereal Research Communications
Authors:
N. Niu
,
Y.X. Bai
,
S. Liu
,
Q.D. Zhu
,
Y.L. Song
,
S.C. Ma
,
L.J. Ma
,
X.L. Wang
,
G.S. Zhang
, and
J.W. Wang

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.

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In this study, a new substitution line, 12-5-1, with 42 chromosomes that was derived from BC3F2 descendants of the hybridization between Triticum aestivum cv. CN19 and Aegilops biuncialis was created and reported. The 12-5-1 was immune to both powdery mildew and stripe rust and has stable fertility. Multi-color fluorescence in situ hybridization indicated that 12-5-1 was a substitution line 1Mb(1B). The seed storage protein electrophoresis showed that 12-5-1 presented high molecular weight glutenin subunits (2 + 12) of CN19 and a new subunit designated as M which apparently originated from parent Ae. biuncialis, and absent 7 + 8 subunits. Additionally, the flour quality parameters showed that the protein content, Zeleny sedimentation value, wet gluten content, and grain hardness and mixing time of 12-5-1 were signifiantly higher than those of its parent CN19. Moreover, 5 pairs of the chromosome 1Mb-specifi polymerase chain reaction-based landmark unique gene markers, TNAC1021, TNAC1026, TNAC1041, TNAC1-02 and TNAC1-04, were also obtained. The new substitution line 1Mb(1B) 12-5-1 could be a valuable source for wheat improvement, especially for wheat end product quality and resistance to disease.

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Cereal Research Communications
Authors:
H.Y. Li
,
Z.L. Li
,
X.X. Zeng
,
L.B. Zhao
,
G. Chen
,
C.L. Kou
,
S.Z. Ning
,
Z.W. Yuan
,
Y.L. Zheng
,
D.C. Liu
, and
L.Q. Zhang

High-molecular-weight glutenin subunits (HMW-GSs) are important seed storage proteins associated with bread-making quality in common wheat (Triticum aestivum L., 2n = 6x = 42, AABBDD). Variation in the Glu-A1x locus in common wheat is scare. Diploid Triticum monococcum ssp. monococcum (2n = 2x = 14, AmAm) is the first cultivated wheat. In the present study, allelic variations at the Glu-A1 m x locus were systematically investigated in 197 T. monococcum ssp. monococcum accessions. Out of the 8 detected Glu-A1 m x alleles, 5 were novel, including Glu-A1 m -b, Glu-A1 m -c, Glu-A1 m -d, Glu-A1 m -g, and Glu-A1 m -h. This diversity is higher than that of common wheat. Compared with 1Ax1 and 1Ax2*, which are present in common wheat, these alleles contained three deletions/insertions as well as some single nucleotide polymorphism variations that might affect the elastic properties of wheat flour. New variations in T. monococcum probably occurred after the divergence between A and Am and are excluded in common wheat populations. These allelic variations could be used as novel resources to further improve wheat quality.

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Cereal Research Communications
Authors:
N. Zhang
,
R.Q. Pan
,
J.J. Liu
,
X.L. Zhang
,
Q.N. Su
,
F. Cui
,
C.H. Zhao
,
L.Q. Song
,
J. Ji
, and
J.M. Li

Plants with deficiency in Gibberellins (GAs) biosynthesis pathway are sensitive to exogenous GA3, while those with deficiency in GAs signaling pathway are insensitive to exogenous GA3. Thus, exogenous GA3 test is often used to verify whether the reduced height (Rht) gene is involved in GAs biosynthesis or signaling pathway. In the present study, we identified the genetic factors responsive to exogenous GA3 at the seedling stage of common wheat and analyzed the response of the plant height related quantitative trait loci (QTL) to GA3 to understand the GAs pathways the Rht participated in. Recombinant inbred lines derived from a cross between KN9204 and J411 with different response to exogenous GA3 were used to screen QTL for the sensitivity of coleoptile length (SCL) and the sensitivity of seedling plant height (SSPH) to exogenous GA3. Two additive QTL and two pairs of epistatic QTL for SCL were identified, meanwhile, two additive QTL and three pairs of epistatic QTL for SSPH were detected. For the adult plant height (PH) investigated in two environments, six additive QTL were identified. Three QTL qScl-4B, qSsph-4B and qPh-4B were mapped in one cluster near the functional marker Rht-B1b. When PH were conditional on SSPH, the absolute additive effect value of qPh-4B and qPh-6B were reduced, suggesting that the Rhts in both two QTL were insensitive to exogenous GA3, while the additive effect values of qPh-2B, qPh-3A, qPh-3D and qPh-5A were not significantly changed, indicating that the Rhts in these QTL were sensitive to exogenous GA3, or they were not expressed at the seedling stage.

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