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The purpose of this study was to investigate the effects of endophytic fungi from tartary buckwheat on the host sprout growth and functional metabolite production. Without obvious changes in the appearance of the sprouts, the exogenous fungal mycelia elicitors notably stimulated the sprout growth and rutin accumulation, and the stimulation effect was mainly depended on the mycelia elicitor species along with its treatment dose. Three endophytic fungi Fat6 (Bionectria pityrodes), Fat9 (Fusarium oxysporum) and Fat15 (Alternaria sp.) were screened to be the most effective candidates for promoting F. tataricum sprout growth and rutin production. With application of polysaccharide (PS, 150 mg/l) of endophyte Fat6, PS (200 mg/l) of endophyte Fat9, and PS (150 mg/l) of endophyte Fat15, the rutin yield was effectively increased to 47.89 mg/(100 sprouts), 45.85 mg/(100 sprouts) and 46.83 mg/(100 sprouts), respectively. That was about 1.5- to 1.6-fold compared to the control culture of 29.37 mg/(100 sprouts). Furthermore, the present study revealed that the biosynthesis of the functional flavonoid resulted from the stimulation of the phenylpropanoid pathway by mycelia polysaccharide treatments. Application of specific fungal elicitors could be an efficient strategy for improving the nutritional and functional quality of tartary buckwheat sprouts.

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Cereal Research Communications
Authors: W. Xue, A. Gianinetti, Y. Jiang, Z. Zhan, L. Kuang, G. Zhao, J. Yan, and J. Cheng

The cereal endosperm provides nutrients for seedling growth. The effects of seed components in seedling establishments under salt stress are, however, not yet fully explored. In this study, 60 barley recombinant inbred lines derived from Lewis × Karl cross were grown in four different environments, and the seed contents of starch, total soluble protein, phytate, total phenolics, total flavonoids and total inorganic phosphorus were determined in the harvested grains. Seeds of each line from the four environments were also assayed for seedling growth under saline treatments from 0 to 400 mM NaCl. Root and shoot lengths after 7 days decreased with increasing salt concentration. Correlations between seed components and either root or shoot length were established across the four seed sources. ANOVA showed a significant environment/source effect for both seed components and seedling growth, although the latter was less affected by the seed-production environment. Modeling seedling length across multiple salinities for each seed source showed that the environment with the most saline-tolerant root-growth curve was that associated the highest seed phosphorus content. Correlations between seed components and seedling growth traits highlighted phytate and total inorganic phosphorus as key components for seedling growth under moderate salinities. Seed phytate contents benefited seedling growth, even at high salinities, suggesting an additional role for this seed component under stressful growth conditions, possibly linked to its potential function as an osmolyte source.

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This study was to examine the effects of four fungal polysaccharides, namely exo-polysaccharide (EPS), water-extracted mycelia polysaccharide (WPS), sodium hydroxideextracted mycelia polysaccharide (SPS), and hydrochloric-extracted mycelia polysaccharide (APS) obtained from the endophytic fungus Bionectra pityrodes Fat6, on the sprout growth and flavonoids production of Fagopyrum tataricum. Without obvious changes in the appearance of the sprouts, the exogenous polysaccharide elicitors notably stimulated the sprout growth and functional metabolites accumulation, and the stimulation effect was mainly depended on the polysaccharide species along with its treatment dose. With application of 150 mg/l of EPS, 150 mg/l of WPS and 200 mg/l of SPS, the total rutin and quercetin yield of buckwheat sprouts was effectively increased to 49.18 mg/(100 sprouts), 50.54 mg/(100 sprouts), and 52.27 mg/(100 sprouts), respectively. That was about 1.57- to 1.66-fold in comparison with the control culture of 31.40 mg/(100 sprouts). Moreover, the present study revealed the accumulation of bioactive flavonoids resulted from the stimulation of the phenylpropanoid pathway by fungal polysaccharide treatments. It could be an efficient strategy for improving the nutritional and functional quality of tartary buckwheat sprouts applied with specific fungal elicitors.

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High molecular weight (HMW) glutenin subunits are important seed storage proteins in wheat and its related species. Novel HMWglutenin subunits in Aegilops tauschii accession of TA2484 were detected and characterized. SDS-PAGE analysis revealed the y-type subunit from TA2484 displayed similar electrophoretic mobility compared to that of 1Dy12 subunit. However, the electrophoretic mobility of x-type subunit was faster than that of 1Dx2 subunit. The primary structure of the two cloned subunits from TA2484 was similar to that of the x- and y-type subunits reported before. However, the 148 residues of the x-type subunit, which contained the sequence element GHCPTSLQQ, in the middle of the repetitive domain was quite different from other x-type subunits. Moreover, the 68 residues in this region were identical to those of the y-type subunits from the same accession. Consequently, 1Dx2.3*t (x-type subunit of TA2484) contains an extra cystenin residue located at the repetitive domain, which is novel compared to the x-type subunits reported so far. Phylogenetic analysis indicated that two subunits from accession TA2484 were in the x- and y-type subunit cluster, but bootstrapping value of 100% gave high support for the spilt between two subunits (1Dx2.3*t and 1Dy12.3*t) and their alleles, respectively. A hypothesis on the genetic mechanism generating this novel sequence of 1Dx2.3*t subunit is suggested.

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High-yield common buckwheat ‘cv. Fengtian 1’ (FT1) and tartary buckwheat ‘cv. Jingqiao 2’ (JQ2) were selected to investigate the characteristics of the grain-filling process and starch accumulation of high-yield buckwheat. FT1 had an average yield that was 43.0% higher than that of the control ‘cv. Tongliaobendixiaoli’ (TLBDXL) in two growing seasons, while JQ2 had an average yield that was 27.3% higher than that of the control ‘cv. Chuanqiao 2’ (CQ2). The Richards equation was utilized to evaluate the grain-filling process of buckwheat. Both FT1 and JQ2 showed higher values of initial growth power and final grain weight and longer linear increase phase, compared with respective control. These values suggest that the higher initial increasing rate and the longer active growth period during grain filling play important roles to increase buckwheat yield. Similar patterns of starch, amylose and amylopectin accumulation were detected in common buckwheat, leading to similar concentration of each constituent at maturity in FT1 and TLBDXL. Tartary buckwheat showed an increasing accumulation pattern of amylose in developing seeds, which differed from that of starch and amylopectin. This pattern led to a significant difference of the concentrations of amylose and amylopectin at maturity between JQ2 and CQ2, the mechanisms of which remained unclear. Nevertheless, both FT1 and JQ2 showed increased starch, amylose, and amylopectin accumulation during the physiological maturity of grains. The results suggest that prolonging the active grain-filling period to increase carbohydrate partitioning from source to seed sink can be an effective strategy to improve buckwheat yield.

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Global rice supplies have been found contaminated with unapproved varieties of genetically modified (GM) rice in recent years, which has led to product recalls in several of countries. Faster and more effective detection of GM contamination can prevent adulterated food, feed and seed from being consumed and grown, minimize the potential environmental, health or economic damage. In this study, a simple, reliable and cost-effective multiplex polymerase chain reaction (PCR) assay for identifying genetic modifications of TT51-1, Kemingdao1 (KMD1) and Kefeng6 (KF6) rice was developed by using the event-specific fragment. The limit of detection (LOD) for each event in the multiplex PCR is approximately 0.1%. Developed multiplex PCR assays can provide a rapid and simultaneous detection of GM rice.

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Male sterile mutants play an important role in the utilisation of crop heterosis. Male sterile plants were found in S5 generations of maize hybrid ZH2, through continuous sib-mating by using the fertile plants in the same population, we obtained a male sterile sibling population K932MS including sterile plants K932S and a fertile plant K932F. The objective of this study was to clarify the genetic characterisation and abortion characteristics by nucleus and cytoplasm effect analyses, cytoplasm grouping, and cytological observation. The results showed that no difference was found between K932S and K932F in the vegetative growth stage, but K932S had no emerging anther or pollen grains. The segregation ratio of fertile plants to sterile plants was 1:1 in the sibling progenies, while it was 3:1 in self-crossing progenies of K932F. The sterility of K932S could be restored among reciprocal progenies when seven normal inbred lines were used as females respectively. The fertility expression of K932S crossed with 30 testers would be changed in different test-crosses and some backcross progenies. The C-type restorer Zifeng-1 (Rf4Rf4) was able to restore the fertility of K932S, and the specific PCR amplification bands of K932MS were consistent with CMSCMo17. The anther of K932S began abortion at dyad with its tapetum expanded radically and vacuolated: this induced abnormality in the shapes of both dyads and tetrads. The microspore could not develop normally, and then it collapsed and gradually disappeared. Hence, K932MS is a C-type cytoplasmic male sterile mutant with a pollen-free, stable inheritance: it has potential application value for further research.

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Physiological male sterility induced by the chemical hybridizing agent (CHA) overcomes problems of maintenance of sterile lines and restorers. However, the mechanism of sterility is unclear. The process of tapetum of CHA-treated ‘Xi’nong 2611’ at uninucleate, binucleate and trinucleate were compared with control to determine if tapetum varying differently during developmental stages. Tapetal degradation in CHA-treated ‘Xi’nong 2611’ began at late uninucleate stage, somewhat earlier than control plants. Cytological observations indicated that the gradual degradation of the tapetum in CHA-treated ‘Xi’nong 2611’ was initiated and terminated earlier than in the control. These findings implied that CHA-induced male sterility was related to abnormally early tapetal degradation. In order to indicate the role of the SKP1 gene in fertility/sterility in wheat, its expression was assessed in anthers at uninucleate, binucleate and trinucleate stages. SKP1 expression was reduced in the later developmental stages, and there was an obvious decrease from the uninucleate to trinucleate stages. Higher expression of the SKP1 gene occurred in ‘Xi’nong 2611’ compared to CHA-treated ‘Xi’nong 2611’. This implied that SKP1 gene expression was inhibited during the fertility transformation process and was related to transformation from fertility to sterility. Moreover, the results from this study suggest that SKP1 plays an essential role of conducting fertility in physiological male sterility.

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To study the development of starch granules in polyploid wheats, we investigated the expression of starch synthetic genes between the synthetic hexaploid wheat SHW-L1, its parents T. turgidum AS2255 and diploid Ae. tauschii AS60. The synthetic hexaploid wheat SHW-L1 showed significantly higher starch content and grain weight than its parents. Scanning electron microscopy (SEM) showed that SHW-L1 rapidly developed starch granules than AS2255 and AS60. The amount of B-type granule in AS60 was less than that in SHW-L1 and AS2255. RT-qPCR result showed that the starch synthetic genes AGPLSU1, AGPLSU2, AGPSSU1, AGPSSU2, GBSSI, SSIII, PHO1 and PHO2 expressed at earlier stages with larger quantity in SHW-L1 than in its parents during wheat grain development. The expression of the above mentioned genes in AS60 was slower than in SHW-L1 and AS2255. The expression pattern of starch synthase genes was also associated with the grain weight and starch content in all three genotypes. The results suggested that the synthetic hexaploid wheat inherited the pattern of starch granule development and starch synthase gene expression from tetraploid parent. The results suggest that tetraploid wheat could plays more important role for starch quality improvement in hexaploid wheat.

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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.

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