RD29A promoter constitutively drives a rice Hd3a expression to promote early-flowering in Saussurea involucrate Kar. et Kir. ex Maxim.

S. involucratae, an endemic and endangered plant, is a valuable and traditional Chinese medicinal herb. In order to control the flowering time of S. involucratae, we used the well-known stress inducible RD29A promoter to drive Hd3a (a FT ortholog from rice) expression in S. involucratae. Unexpectedly, the majority of regenerated buds in RD29A::Hd3a transgenic lines (S-RH) produced flowers in tissue culture stage under normal growth (25 ± 2 °C) condition. Their flowering time was not further influenced by salt treatment. Hd3a in S-RH was strongly expressed in MS media supplemented with or without 50 mM NaCl. RD29A::GUS transgenic experiments further revealed that RD29A constitutively promoted GUS expression in both S. involucrate and halophyte Thellungiella halophile, in contrast to glycophic plants Oryza sativa L. 'Zhonghua 11', in which its expression was up-regulated by cold, salinity, and drought stress. The results supported the hypothesis that RD29A promoter activity is inducible in stress-sensitive plants, but constitutive in stress-tolerant ones. Importantly, S-RH plants produced pollen grains and seeds under normal conditions. Additionally, we found that OsLEA3-1::Hd3a and HSP18.2::Hd3a could not promote S. involucrate to flower under either normal conditions or abiotic stresses. Taken together, we demonstrated the potential of RD29A::Hd3a might be served as a feasible approach in breeding S. involucrate under normal condition.

Pyramiding of gn1a, gs3, and ipa1 Exhibits Complementary and Additive Effects on Rice Yield.

Pyramiding of quantitative trait loci (QTLs) is a powerful approach in breeding super-high-yield varieties. However, the performance of QTLs in improving rice yield varies with specific genetic backgrounds. In a previous study, we employed the CRISPR/Cas9 system to target three yield-related genes, gn1a, gs3, and ipa1 in japonica 'Zhonghua 11', mutants of which featured large panicle, big grain, few sterile tillers, and thicker culm, respectively. In this paper, four pyramided lines, including gn1a-gs3, gn1a-ipa1, gs3-ipa1, and gn1a-gs3-ipa1, were further generated by conventional cross-breeding to be tested. Agronomic traits analysis showed that: (1) the stacking lines carried large panicles with an increased spikelet number in the main panicle or panicle; (2) the grain weight of the stacking lines, especially gs3-ipa1 and gn1a-gs3-ipa1, were heavier than those in single mutants; (3) both gn1a-gs3 and gs3-ipa1 produced more grain yield per plant than single mutant lines; (4) pyramided lines were higher than single mutants and transcriptome analysis found improved expression levels of genes related to lipid, amino acid, and carbohydrate transport and metabolism in lines pyramiding three mutant alleles, possibly as a result of complementary and additive effects. Accordingly, the alteration of gene-expression patterns relating to hormone signaling, plant growth, and seed size control was characterized in pyramided lines. The present study not only investigates the effects of pyramiding genes, but also may provide an efficient strategy for breeding super-high-yield rice by reducing the time cost of developing pyramided lines.

[Review on the assessment model of nutrient recycling with agricultural residues treatment technologies]. / å†œä¸šåºŸå¼ƒç‰©å »åˆ†å¾ªçŽ¯åˆ©ç”¨æŠ€æœ¯æ¨¡å¼è¯„ä¼°æ¨¡åž‹çš„ç ”ç©¶è¿›å±•.

The technology for nutrient resource utilization of agricultural residues is crucial to realizing a circular agricultural economy. The assessment model provides essential support to optimize nutrient recovery and treatment technologies. We specifically summarized research progress in the assessment framework of agricultural residues nutrient recycling technology, assessment models and evaluation indicators, data sources of models and their uncertainty analysis, and the application scale of models. We found that process mathematical models and industrial eco-logy models are the common nutrient flow assessment models. Process mathematical and industrial ecology models differed greatly in terms of the reliability of assessment results and the simulation scale. The former mainly focused at laboratory or pilot scale, with higher accuracy. The latter could achieve multi-scale simulation from microscopic to macroscopic and had higher uncertainty, due to the way its data were obtained. Finally, we provided an outlook on the research on the assessment model of agricultural residues nutrient resource utilisation technology. In order to achieve accurate assessment of waste resource utilisation technology in agricultural production systems at the regional scale, a reliable model framework and database should be established by combining process mathematical models with industrial ecology models. Meanwhile, we should carry out research on model expansion at the geographical scales of factory scale, farm scale, village scale, township scale, and regional scale.

A new species of early-diverging Sauropodiformes from the Lower Jurassic Fengjiahe Formation of Yunnan Province, China.

Sauropodomorpha were herbivorous saurischian dinosaurs that incorporate Sauropoda and early-diverging sauropodomorphs. The oldest sauropodomorph remains are known from Late Triassic deposits, most of them Gondwanan. The Laurasian record comprises some Triassic forms, but the bulk is Jurassic in age. Among the 14 Jurassic non-sauropodan sauropodomorphs from Laurasia described in the past, 8 are from China. Here we describe a new non-sauropodan sauropodomorph, Irisosaurus yimenensis gen. et sp. nov., from the Early Jurassic Fengjiahe Formation of China. Nearly all of the non-sauropodan sauropodomorph genera currently known from China were first reported from the Lufeng Formation. The Fengjiahe Formation is its Southern equivalent, bringing a fauna similar to that of the Lufeng Formation to light. The new genus is defined based on an incomplete but unique maxilla, with a premaxillary ramus higher than long prior to the nasal process, a large and deep neurovascular foramen within the perinarial fossa, and a deep perinarial fossa defined by a sharp rim. Phylogenetic analysis places Irisosaurus at the very base of Sauropodiformes, as the sister-taxon of the Argentinean genus Mussaurus. This specimen adds to a growing assemblage of Chinese Jurassic non-sauropodan sauropodomorphs that offers new insight into the Laurasian evolution of this clade.

[Efficient identification of gene knockout mutant mediated by CRISPR/Cas9 by CELâ crude extracts].

CRISPR/Cas9, emerged as an efficient and powerful gene editing technology, has become the mainstream genome editing technology. Constructing mutants using CRISPR/Cas9 system is of great significance to the functional study and breeding application of useful genes. As the basis of the technology, a method for identification of mutation with efficiency and lower cost is needed. In this report, we studied the factors influencing mutation detected by CEL Ⅰ crude extracts, such as the amount of protein, enzyme incubation time, PCR buffers. Under the optimized conditions, we can integrate the mutation detection steps into one-tube reaction. We used this system to examine the mutation types and frequency of rice stn1 mediated by CRISPR/Cas9. We also used this method to identify different mutation types including homozygous, heterozygous and bi-allelic mutations. The accuracy of this method reached 100% verified by sequencing. Altogether, our results showed that using CELⅠ crude extracts was an efficient and low cost method for identification of CRISPR/Cas9 mediated mutation.

Screening of Proximal and Interacting Proteins in Rice Protoplasts by Proximity-Dependent Biotinylation.

Proximity-dependent biotin identification (BioID), which detects physiologically relevant proteins based on the proximity-dependent biotinylation process, has been successfully used in different organisms. In this report, we established the BioID system in rice protoplasts. Biotin ligase BirAG was obtained by removing a cryptic intron site in the BirA∗ gene when expressed in rice protoplasts. We found that protein biotinylation in rice protoplasts increased with increased expression levels of BirAG. The biotinylation effects can also be achieved by exogenous supplementation of high concentrations of biotin and long incubation time with protoplasts. By using this system, multiple proteins were identified that associated with and/or were proximate to OsFD2 in vivo. Our results suggest that BioID is a useful and generally applicable method to screen for both interacting and neighboring proteins in their native cellular environment in plant cell.

Effect of high-dose rosuvastatin loading before percutaneous coronary intervention in Chinese patients with acute coronary syndrome: A systematic review and meta-analysis.

BACKGROUND: Acute coronary syndrome (ACS) is an important disease threatening human life and health. Many studies have shown that the loading dose of atorvastatin can significantly improve the prognosis of patients with ACS, and reduce the mortality. However, this conclusion is not consistent. Thus, we aimed to evaluate the effect of high-dose rosuvastatin loading before percutaneous coronary intervention (PCI) in Chinese patients with ACS using a meta-analysis based on a systematic review of published articles. METHODS: We systematically reviewed published studies, evaluating the effect of high-dose rosuvastatin loading before percutaneous coronary intervention in Chinese patients with ACS. The retrieval time is limited from inception to 2 November 2016, and the retrieved databases included PubMed, Embase, the Cochrane Library, Web of Science, CBM, CNKI, the VIP database and the Wang Fang database. Two researchers independently assessed the quality of the included studies and then extracted the data. Stata 11.0 was used for data analysis. RESULTS: In total, 11 articles, which included 802 patients, were included in our meta-analysis. Among these patients, 398 patients were in the high-dose group (20 mg/day) and 404 patients were in the conventional dose group (10 mg/day). Meta-analysis results showed that compared with the conventional dose group: 1) The loading dose of rosuvastatin can significantly reduce the hs-CRP level after PCI, including at 24 hours (SMD = -0.65, 95%CI -0.84 ~ -0.47, P = 0.000), 48 hours (SMD = -0.40, 95%CI -0.68 ~ -0.11, P = 0.006), and four weeks (SMD = -1.64, 95%CI -2.01 ~ -1.26, P = 0.000). 2) The loading dose of rosuvastatin can significantly reduce the levels of LDL-C and cTnT, including the level of LDL-C at 30 d after PCI (SMD = -0.89, 95%CI -1.10 ~ -0.69, P = 0.000), and the level of cTnT at 24 h after PCI (SMD = -1.93, 95%CI -2.28 ~ -1.59, P = 0.000), and increase the level of HDL-C at 48 h after PCI (SMD = 0.61, 95%CI 0.34 ~ 0.88, P = 0.000). 3) The loading dose of rosuvastatin can significantly reduce the levels of TG and TC, including the level of TG at 30 d after PCI (SMD = -0.94, 95%CI -1.17 ~ -0.71, P = 0.000), the level of TC at 48 h after PCI (SMD = -0.35, 95%CI -0.68 ~ -0.01, P = 0.043), and the level of TC at 30 d after PCI (SMD = -0.77, 95%CI -0.98 ~ -0.56, P = 0.000). CONCLUSIONS: Our systematic review and meta-analysis showed that, compared with the conventional dose, the loading dose of rosuvastatin was more beneficial to patients with ACS in China and is suitable for clinical application. Due to the limitations of the quality and quantity of included articles, this conclusion still needs to be confirmed by multicenter clinical trials.

A rice DEAD-box RNA helicase protein, OsRH17, suppresses 16S ribosomal RNA maturation in Escherichia coli.

DEAD-box proteins comprise a large protein family. These proteins function in all types of processes in RNA metabolism and are highly conserved among eukaryotes. However, the precise functions of DEAD-box proteins in rice physiology and development remain unclear. In this study, we identified a rice DEAD-box protein, OsRH17, that contains a DEAD domain and all of the common conserved motifs of DEAD-box RNA helicases. OsRH17 was specifically expressed in pollen and differentiated callus and upregulated by application of the plant hormones naphthyl acetic acid (NAA) and abscisic acid (ABA). The OsRH17:GFP fusion protein was localized to the nucleus. Tiny amounts of OsRH17 and partial fragments (N-427 and C-167) were detected when they were expressed in Escherichia coli, a prokaryote. Growth of the host cells was suppressed in E. coli by OsRH17, N-427 or C-167, and this suppression was independent of the concentration of the NaCl in the medium. Expression analysis of rRNAs in E. coli revealed that the 16S rRNA precursor accumulated in transgenic E. coli cells, and the relative growth rate was inversely proportional to the levels of pre-16S rRNA accumulation. Results suggested that OsRH17 may play a role in ribosomal biogenesis and suppress 16S rRNA maturation in E. coli. No visible phenotype was observed in transgenic yeast and rice (overexpressing OsRH17, N-427, and C-167, as well as OsRH17 knockdown), and even in some abiotic and biotic stresses, which could be due to the redundancy in rice under normal conditions.

An effective method of RNA isolation from Fallopia multiflora tuberous roots.

To isolate high-quality total RNA from Fallopia multiflora tuberous roots is difficult because of the presence of high levels of carbohydrates, phenolics, and other secondary metabolites. Since several procedures specialized for RNA isolation from polysaccharides and phenols rich tissues have resulted in poor yields, in this study, we developed a modified protocol that was derived from the traditional CTAB method. The protocol was able to produce high-quality and intact RNA from the tuberous roots of F. multiflora. The yield of total RNA was more than 0.15 mg/g fresh weight, with an A260/A280 ratio of 1.9-2.0. The obtained RNA was of sufficient quality and suitable for downstream application such as reverse-transcription polymerase chain reaction (RT-PCR), Northern hybridization, and cDNA library construction. The protocol may also have wider applicability for total RNA isolation from other plant species with tuberous roots.

[Pollination ecology of cultivated Ficus pumila var. awkeotsang].

The study on the pollination ecology of cultivated Ficus pumila var. awkeotsang showed that this plant had two apparent blossoming periods, i.e., in spring and autumn, which were relatively longer than those of the wild one. Cultivated Ficus pumila var. awkeotsang couldnt rely on the wasps living in the variant syconia to pollinate or lay eggs. Under the circumstances the wasps were separated from wild F. pumila var. awkeotsang, the pollination system was damaged because of the absence of the wasps. The pollination wasps could be introduced artificially to cultivated F. pumila var. awkeotsang, but the pollination system was restored relatively slowly. The rate of syconia hanging on trees could only reach 51.11% after four years, with an average production rate of the female flowers being 64.37%, and an average gall tuming rate of gall flowers being 64.35%. The growing syconia degree of female and male flowering phases was low, and the collective way of the wasps' flying out and the small number of the wasps entering the syconia were the causes of low rate co-existing breeding.