This study characterized the effect of green manures (February orchid, hairy vetch, rattail fescue and a no-green-manure control) and the termination method (flail or disk) on nutrient contents, enzyme activities, microbial biomass, microbial community structure of rhizosphere soil and vegetative growth of walnut tree. All three selected green manures significantly enhanced the water content, organic C, total N and available P. The rattail fescue significantly decreased the mineral N. Total organic C, total N, mineral N and available P were significantly greater under flail than under disk. Hairy vetch and February orchid significantly improved levels of soil β-glucosidase, N-acetyl-glucosaminidase and acid phosphatase activity, whereas rattail fescue improved only β-glucosidase activity. All of the green manures significantly decreased phenoloxidase activity. β-glucosidase, N-acetyl-glucosaminidase and acid phosphatase activities were significantly greater under flail relative to disk. The termination method had no significant effect on phenoloxidase activity. The different types of green manures and termination methods significantly altered the soil microbial biomass and microbial community structure. The green-manure treatments were characterized by a significantly greater abundance of Gram-positive (Gram +) bacteria, total bacteria and saprophytic fungi compared to the control. Hairy vetch significantly decreased the abundance of arbuscular mycorrhizal fungi (AMF) while February orchid and rattail fescue increased their abundance compared to the no-green-manure treatment. The abundance rates of Gram+ bacteria, actinomycetes, saprophytic fungi and AMF were significantly greater in soils under flail than under disk. In terms of vegetative growth of walnut tree, hairy vetch showed the greatest positive effects. The growth of walnut tree was significantly greater under flail relative to disk. Our results indicate that green-manure application benefits the rhizosphere soil micro-ecology, rhizosphere soil nutrient contents and tree growth. Overall, the hairy vetch and flail combined treatment is recommended for walnut orchards in northern China.
In this study, a novel composite hydrogel with improved cellular structure and mechanical properties was prepared by the crosslinking of hyaluronic acid (HA) and sodium alginate (SAL). The amide linkages (covalent bonds) in the hydrogel that we expected to form were confirmed by Fourier transform infrared spectroscopy. The hydrogels had a pore size larger than 100 lm and were observed by scanning electron microscopy. Texture profile analysis indicated that the hardness of the hydrogels was enhanced by an increase in the polymer's concentration, but it declined with an increase in the HA/SAL molar ratio. The swelling capacity was reduced with increases in the polymer's concentration and the 1-ethyl-3-(3-dimethyl aminopropyl)-1-carbodiimide hydrochloride (EDC)/HA molar ratio, and it was enhanced by an increase in the HA/SAL molar ratio. The resistance against hyaluronidase was negatively correlated with the proportion of HA in the hydrogels and positively correlated with the EDC/HA molar ratio. Given the improved physicochemical properties that we produced, these novel hydrogels may have the potential to be applied in tissue engineering scaffolding.
Hawthorns (Crataegus L.) are one of the most important processing and table fruits in China, due to their medicinal properties and health benefits. However, the interspecific relationships and evolution history of cultivated Crataegus in China remain unclear. Our previously published data showed C. bretschneideri may be derived from the hybridization of C. pinnatifida with C. maximowiczii, and that introgression occurs between C. hupehensis, C. pinnatifida, and C. pinnatifida var. major. In the present study, chloroplast sequences were used to further elucidate the phylogenetic relationships of cultivated Crataegus native to China. The chloroplast genomes of three cultivated species and one related species of Crataegus were sequenced for comparative and phylogenetic analyses. The four chloroplast genomes of Crataegus exhibited typical quadripartite structures and ranged from 159,607 bp (C. bretschneideri) to 159,875 bp (C. maximowiczii) in length. The plastomes of the four species contained 113 genes consisting of 79 protein-coding genes, 30 tRNA genes, and 4 rRNA genes. Six hypervariable regions (ndhC-trnV(UAC)-trnM(CAU), ndhA, atpH-atpI, ndhF, trnR(UCU)-atpA, and ndhF-rpl32), 196 repeats, and a total of 386 simple sequence repeats were detected as potential variability makers for species identification and population genetic studies. In the phylogenomic analyses, we also compared the entire chloroplast genomes of three published Crataegus species: C. hupehensis (MW201730.1), C. pinnatifida (MN102356.1), and C. marshallii (MK920293.1). Our phylogenetic analyses grouped the seven Crataegus taxa into two main clusters. One cluster included C. bretschneideri, C. maximowiczii, and C. marshallii, whereas the other included C. hupehensis, C. pinnatifida, and C. pinnatifida var. major. Taken together, our findings indicate that C. maximowiczii is the maternal origin of C. bretschneideri. This work provides further evidence of introgression between C. hupehensis, C. pinnatifida, and C. pinnatifida var. major, and suggests that C. pinnatifida var. major might have been artificially selected and domesticated from hybrid populations, rather than evolved from C. pinnatifida.
Hawthorn (Crataegus pinnatifida var. major), a cultivated fruit tree, is native and unique to China. Its fruits have high nutritional, health, and medicinal values. However, the development and ripening process of hawthorns is accompanied by dramatic changes in flavor, aroma, and bioactive phytochemicals, which are the fundamental factors that contribute to the potential health benefits and establishment of fruit quality. Therefore, an exploration of the dynamic changes in metabolites and their regulatory networks during the development of hawthorn fruits can elucidate the formation mechanisms of active substances in hawthorn fruits. In this study, we used a broad targeted metabolomics approach to identify and analyze the dynamics of metabolites in hawthorn fruits at five developmental stages. The results revealed 998 primary and secondary metabolites that were classified into 15 categories. The accumulation levels of most sugars increased during fruit development and then accelerated at the fruit ripening stage. The accumulation levels of a few organic acids (e.g., citric acid, isocitric acid, and quinic acid) continuously increased. Many organic acids exhibited significant decreasing trends. Among the 561 secondary metabolites detected, 189 were phenolic acids and 199 were flavonoids. The levels of many flavonoids were significantly reduced at later stages of fruit development; in contrast, the levels of two anthocyanins significantly increased during fruit ripening. Correlation analysis revealed that there is a certain correlation within and between primary as well as secondary metabolites during fruit development. Furthermore, the integration of metabolomic and transcriptomic data in this study revealed that changes in the expression of some differentially expressed genes (DEGs) were associated with the accumulation of metabolites such as sugars, organic acids, and flavonoids, e.g., the upregulated expression levels of CS (citrate synthase) genes were consistent with the continued accumulation of citric acid. Overall, this study demonstrates the metabolic changes that occur during the development of hawthorn fruit, explores the molecular mechanisms that underlie metabolite changes during fruit development, and lays a strong theoretical foundation for the improvement of hawthorn fruit quality and the development of functional components.
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