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Abstract Background Agrobacterium rhizogenes-mediated (ARM) transformation is a highly efficient technique for generating composite plants composed of transgenic roots and wild-type shoot, providing a powerful tool for studying root biology. The ARM transformation has been established in many plant species, including soybean. However, traditional transformation of soybean, transformation efficiency is low. Additionally, the hairy roots were induced in a medium, and then the generated composite plants were transplanted into another medium for growth. This two-step operation is not only time-consuming, but aggravates contamination risk in the study of plant-microbe interactions. Results Here, we report a one-step ARM transformation method with higher transformation efficiency for generating composite soybean plants. Both the induction of hairy roots and continuous growth of the composite plants were conducted in a single growth medium. The primary root of a 7-day-old seedling was decapitated with a slanted cut, the residual hypocotyl (maintained 0.7-1 cm apical portion) was inoculated with A. rhizogenes harboring the gene construct of interest. Subsequently, the infected seedling was planted into a pot with wet sterile vermiculite. Almost 100% of the infected seedlings could produce transgenic positive roots 16 days post-inoculation in 7 tested genotypes. Importantly, the transgenic hairy roots in each composite plant are about three times more than those of the traditional ARM transformation, indicating that the one-step method is simpler in operation and higher efficiency in transformation. The reliability of the one-step method was verified by CRISPR/Cas9 system to knockout the soybean Rfg1, which restricts nodulation in Williams 82 (Nod-) by Sinorhizobium fredii USDA193. Furthermore, we applied this method to analyze the function of Arabidopsis YAO promoter in soybean. The activity of YAO promoter was detected in whole roots and stronger in the root tips. We also extended the protocol to tomato. Conclusions We established a one-step ARM transformation method, which is more convenient in operation and higher efficiency (almost 100%) in transformation for generating composite soybean plants. This method has been validated in promoter functional analysis and rhizobia-legume interactions. We anticipate a broad application of this method to analyze root-related events in tomato and other plant species besides soybean.

Stellera chamaejasme is one of the main poisonous weeds distributed in alpine meadow of Qinghai Province. Rapid spreading of S. chamaejasme has done serious harm to local animal husbandry and caused continuous grassland ecosystem degradation. This paper focused on the spectral differences between S. chamaejasme and herbage, taking the typical degraded alpine meadow dominated by S. chamaejasme in Qilian County of Haibei Region as the test site and using the spectral measurements acquired in the full?blossom period of S. chamaejasme from 2012 to 2014. The results showed that the spectral behavior of flowers of S. chamaejasme differed significantly from green background that included leaves of S. chamaejasme and herbage within 350–900 nm of VIS-NIR wavebands. The biggest reflectance difference between flowers of S. chamaejasme and green background was located in the red valley, followed by the blue valley. The reflectance of S. chamaejasme community increased with the rising of coverage, the biggest reflectance difference between S. chamaejasme and herbage communities lied in the near-infrared peak, and the best separability between S. chamaejasme communities with different coverage was also at the point. The difference of first derivative spectra between flowers of S. chamaejasme and green background located in amplitude of yellow edge was remarkable, followed by amplitude of blue edge, the same as differences between S. chamaejasme and herbage communities. Linear regression analysis between coverage of S. chamaejasme and spectral feature parameters showed best result for red valley (R2=0.94). Finally, the red valley, the blue valley and the near-infrared peak were proposed for discriminating S. chamaejasme from herbage in the full-blossom period of S. chamaejasme, and the combination of corresponding red, blue and near-infrared bands could be used to build sensitive indices for S. chamaejasme recognition. [ABSTRACT FROM AUTHOR]

Large-scale commercial bioprocesses that manufacture biopharmaceutical products such as monoclonal antibodies generally involve multiple bioreactors operated in parallel. Spectra recorded during in situ monitoring of multiple bioreactors by multiplexed fiber-optic spectroscopies contain not only spectral information of the chemical constituents but also contributions resulting from differences in the optical properties of the probes. Spectra with variations induced by probe differences cannot be efficiently modeled by the commonly used multivariate linear calibration models or effectively removed by popular empirical preprocessing methods. In this study, for the first time, a calibration model is proposed for the analysis of complex spectral data sets arising from multiplexed probes. In the proposed calibration model, the spectral variations introduced by probe differences are explicitly modeled by introducing a multiplicative parameter for each optical probe, and then their detrimental effects are effectively mitigated through a "dual calibration" strategy. The performance of the proposed multiplex calibration model has been tested on two multiplexed spectral data sets (i.e., MIR data of ternary mixtures and NIR data of bioprocesses). Experimental results suggest that the proposed calibration model can effectively mitigate the detrimental effects of probe differences and hence provide much more accurate predictions than commonly used multivariate linear calibration models (such as PLS) with and without empirical data preprocessing methods such as orthogonal signal correction, standard normal variate, or multiplicative signal correction. [ABSTRACT FROM AUTHOR]

The quantitative analysis of spatio-temporal variations of vegetation cover and its correlation with climate are of great significance for understanding of ecological environment, ecological civilization construction, and sustainable development in semi-arid areas. We investigated the spatio-temporal variations of normalized difference vegetation index (NDVI) and its response to climate change during 2000-2020 in Xilin Gol, Inner Mongolia, by using trend analysis, regression analysis and partial correlation analysis based on the data of MODIS-NDVI, tempe-rature, precipitation, digital elevation model. The results showed that vegetation cover in Xilin Gol had been increased from 2000 to 2020, which generally included three phases, i.e. , stable fluctuation, rapid growth, and steady growth. The mean NDVI showed a zonal increasing distribution from southwest to northeast, and had a strong correlation with elevation and population density in Xilin Gol region. The high values of NDVI were mainly in the east, with a significant increasing trend, and the low values were in the southwest, with a local degradation. The sensitivity of vegetation cover to climate change showed spatial and temporal variations. The spatial variation of vegetation was more sensitive to temperature and the interannual variation was sensitive to annual precipitation. In summary, vegetation cover improved overall in Xilin Gol, but there was degradation in some areas. We should formulate differentiated and precise vegetation restoration and ecological environmental protection policies.

Virtual ecological technology is a new technology that builds the foundation of a digital ecological system, simulates the process of ecological evolution, and establishes a global knowledge analysis system based on a unified spatio-temporal benchmark. It is a new direction for the interdisciplinary integration and development of eco-logy, geographic information science, computer science and other subjects towards modernization, informatization, and intelligence. Research, development, and application of virtual ecological technology is of great significance for the development of ecology as a discipline, ecosystem management, and regional sustainable development. Curren-tly, research on virtual ecological technology is still in its infancy and lacks a complete and clearly defined framework, making it difficult to support systematic iterative development and scientific analysis. In this paper, starting from the main theories and objectives of modern ecology, we summarized the main contents and technical requirements of virtual ecological construction, and proposed a key technical system of virtual ecology that integrated vir-tual geography and digital twin technology. From the perspective of application scenarios, we analyzed the application capabilities of virtual ecological technology in air pollution and energy analysis. Finally, we summarized the deve-lopment potential of virtual ecological technology in the digital construction of the ecological environment, intelligent computing, and realistic expression of different levels of ecological space resources transformation under the background of new generation of information technology, both at the local computing point breakthrough and the global technology point fusion aspects. This would enrich and advance the technical capabilities that support China's ecological space resource transformation.

Stellera chamaejasme is one of the most serious weeds in Qinhai-Tibetan Plateau, the rapid expansion of which exerts an increasing effect on the alpine meadow ecosystem. With high-throughput sequencing technology, geostatistics and GIS method, the spatial heterogeneity of soil fungal diversity in Stellera occurrence area and the spatial correlation between Stellera coverage and soil fungal diversity were investigated in a typical degraded alpine meadow of the Qilian Mountain. Compared to no-Stellera area, the fungi richness in Stellera area decreased, the dominance increased, and the α-diversity reduced. The difference of fungal species composition enhanced and β-diversity significantly increased. The spatial pattern of soil fungal diversity was affected by the invasion of Stellera, resulting in higher fragmentation in occurrence area. Spatial heterogeneity of species composition increased remarkably, and spatial stability of α-diversity and β-diversity decreased. The portion of positive correlation and negative correlation interlaced, indicating no clear spatial correlation between Stellera coverage and soil fungal diversity. Our results indicate that the spatial pattern of soil fungal diversity was affected by the interaction of soil and vegetation in Stellera invaded meadows.

Quantitative evaluation on the vegetation recovery after the earthquake is of great scienti-fic significance for local ecological system rebuilding and regional social-economic sustainable deve-lopment. By taking the Longxi River Basin in Dujiangyan of Sichuan Province as an example, and employing the MODIS-NDVI time-series data, this paper illustrated the dynamic changes of the ve-getation cover rates in Longxi River basin before and after the 5.12 Wenchuan Earthquake by using Landsat data, and then time-space characters of the recovery rates of the damaged vegetation was quantified structurally by referring the factors of river-system and terrain. The results showed that the recovery of impaired vegetation coverage worked well from a holistic point of view, but the response of vegetation coverage to earthquake disaster showed a lagging-phenomenon. The recovery rate of damaged vegetation was significantly correlated to its distance to surrounding river systems, elevation, slope degree, and slope aspect. This finding could provide technical supports for decision-makers to intervene the recovering process after earthquake.

Quantifying plant litter cover is important to evaluate the effectiveness of vegetation to protect soil against soil erosion. Field campaign was carried out in the Loess Hilly-gully Region of Northern Shaanxi to acquire the spectral reflectance data of plant-soil mixed scenes for two broadleaf forests and rehabilitated grassland. Spectral behavior of plant-soil mixed scenes was analyzed and the effectiveness of hyperspectral indexes NDLI (normalized difference lignin index) and CAI (cellulose absorption index) for quantifying plant litter cover was evaluated based on lab and field experiments. The results showed spectra reflectance of plant-soil mixed scenes with different proportions of litter and soil varied greatly by the influence of color and water content. The NDLI and CAI values increased with the rise of proportion of litter within the mixed scene under dry and wet status, regression analysis between the two indexes and plant litter cover of mixed scenes showed that the obviously better result for CAI (R2=0.98, rehabilitated grassland and broadleaf forests under dry status). The discrimination between mixed scenes and soil using CAI significantly outperformed NDLI under dry status; the estimation of plant litter cover by CAI is more effective compared to NDLI. Lab analysis was validated by field measuring: R2=0.90 showed highest correlation between CAI and plant litter cover for rehabilitated grassland. The validity of estimation of plant litter cover by both two indexes reduced to some degree in the field. The research enhanced the scientific basis for remote sensing estimation of plant litter cover.

Stellera chamaejasme is one of the main poisonous weeds distributed in alpine meadow of Qinghai Province. Rapid spreading of S. chamaejasme has done serious harm to local animal husbandry and caused continuous grassland ecosystem degradation. This paper focused on the spectral differences between S. chamaejasme and herbage, taking the typical degraded alpine meadow dominated by S. chamaejasme in Qilian County of Haibei Region as the test site and using the spectral measurements acquired in the full-blossom period of S. chamaejasme from 2012 to 2014. The results showed that the spectral behavior of flowers of S. chamaejasme differed significantly from green background that included leaves of S. chamaejasme and herbage within 350-900 nm of VIS-NIR wavebands. The biggest reflectance difference between flowers of S. chamaejasme and green background was located in the red valley, followed by the blue valley. The reflectance of S. chamaejasme community increased with the rising of coverage, the biggest reflectance difference between S. chamaejasme and herbage communities lied in the near-infrared peak, and the best separability between S. chamaejasme communities with different coverage was also at the point. The difference of first derivative spectra between flowers of S. chamaejasme and green background located in amplitude of yellow edge was remarkable, followed by amplitude of blue edge, the same as differences between S. chamaejasme and herbage communities. Linear regression analysis between coverage of S. chamaejasme and spectral feature parameters showed best result for red valley (R2 = 0.94). Finally, the red valley, the blue valley and the near-infrared peak were proposed for discriminating S. chamaejasme from herbage in the full-blossom period of S. chamaejasme, and the combination of corresponding red, blue and near-infrared bands could be used to build sensitive indices for S. chamaejasme recognition.

Objective: To prepare the SEA-TM and mB7.1-GPI dual-anchored EL-4 cell vaccine and to investigate its antitumor effects., Methods: mB7.1-GPI-anchored EL-4 cell vaccine, SEA-TM-anchored EL-4 cell vaccine, SEA-TM and mB7.1-GPI dual-anchored EL-4 cell vaccine were prepared. In vitro the biological activities of these vaccines were measured using a lymphocyte proliferation assay and cytokine release assay on splenocytes derived from C57BL/6 mice. The splenocytes were co-cultured with EL-4 or EL-4/mB7.1-GPI or EL-4/SEA-TM or EL-4/SEA-TM + mB7.1-GPI (treated with Mitomycin C). Lymphocyte proliferation was determined with MTT assay, the concentrations of cytokines (IL-2 and IFN-gamma) were measured using a ELISA technique. Forty C57BL/6 mice were inoculated with EL-4 cells, after 3 days the mice were randomly divided into 5 groups with 8 in each and were treated with PBS, EL-4 cell vaccine, EL-4/mB7.1-GPI cell vaccine, EL-4/SEA-TM cell vaccine and EL-4/SEA-TM + mB7.1-GPI cell vaccine respectively, vaccines were injected three time with two-day interval. Animals were observed daily, tumor sizes were measured every third day. Twenty-five days after tumor challenge, 3 mice in each group were sacrificed and splenic lymphocytes were isolated to examine the activity of natural killer cells (NK) and cytolytic T lymphocytes (CTL). The survival of the remaining 5 mice in each group was observed till the 90th day., Results: mB7.1-GPI or/and TM-SEA fusion protein was stably anchored onto the surface of EL-4 tumor cells. EL-4/mB7.1-GPI or EL-4/SEA-TM had a stronger ability to stimulate lymphocyte proliferation and IL-2 and IFN-gamma production than EL-4 (P < 0.05); while EL-4/SEA-TM + mB7.1-GPI showed a further increased ability than EL-4/mB7.1-GPI and EL-4/SEA-TM in stimulating lymphocyte proliferation and cytokine production in vitro (P < 0.05). Volume of tumor was smaller and survival time of mice was longer in EL-4/mB7.1-GPI vaccine group, EL-4/SEA-TM vaccine group and EL-4/SEA-TM + mB7.1-GPI vaccine group, comparing with PBS group and EL-4 cell vaccine group (P < 0.05). Tumor volume was much smaller and survival time of mice was much longer in EL-4/mB7.1-GPI + mB7.1-GPI vaccine group, comparing with EL-4/SEA-TM vaccine group and EL-4/mB7.1-GPI vaccine group (P < 0.05). Lymphocytes derived from the mice treated with EL-4/SEA-TM + mB7.1-GPI showed much higher NK activity and CTL activity than those derived from EL-4/mB7.1-GPI vaccine group and EL-4/SEA-TM vaccine group (P < 0.05), meanwhile the NK activity and CTL activity of EL-4/mB7.1-GPI vaccine group and EL-4/SEA-TM vaccine group was higher than EL-4 vaccine group (P < 0.05)., Conclusion: mB7.1-GPI or/and SEA-TM fusion protein was stably anchored onto the surface of EL-4 tumor cells. The tumor cell vaccines prepared from these cells exhibited antitumor effect. The mB7.1-GPI and SEA-TM dual-anchored tumor cell vaccine had much stronger antitumor effect than the single-anchored tumor cell vaccine.

Objective: To investigate the influencing factors of polyethylenimine (PEI) in gene transfer in vitro., Methods: Cytotoxic effects of PEI on in vitro cultured NIH 3T3 cells were quantified by MTT assay. The interaction between PEI and DNA at different charge ratios was analyzed by agarose gel electrophoresis retardation assay. The expression of gene transfer was monitored in Cos-7 cells using pEGFP and pSV beta plasmids as the reporter gene systems. Influences of chloroquine, albumin, serum, salt ion strength, and Mg(2+) ion and other factors on PEI/DNA transfer efficiency were evaluated., Result: The survival rate of NIH3T3 cells at 6 mg/L of PEI was 64.2% and at 7 mg/L of PEI was 54.4%. Gel electrophoresis retardation assays showed that PEI completely retarded DNA migration at 3.0 PEI nitrogen per DNA phosphate. Chloroquine enhanced the transfection efficiency of PEI. Albumin and serum in the culture medium decreased the transfection efficiency. HBS(HEPES buffered solution) or 150 mmol/L NaCl as the dilution solution of PEI/DNA was superior over 278 mmol/L glucose solution in the transfection efficiency. Mg(2+) in the dilution solution decreased the transfer efficiency of PEI/DNA., Conclusion: PEI is efficient gene transfer agent of eukaryotes in vitro, and can be possibly used in vivo.