Highly efficient Agrobacterium rhizogenes-mediated hairy root transformation in citrus seeds and its application in gene functional analysis.

Highly efficient genetic transformation technology is beneficial for plant gene functional research and molecular improvement breeding. However, the most commonly used Agrobacterium tumefaciens-mediated genetic transformation technology is time-consuming and recalcitrant for some woody plants such as citrus, hampering the high-throughput functional analysis of citrus genes. Thus, we dedicated to develop a rapid, simple, and highly efficient hairy root transformation system induced by Agrobacterium rhizogenes to analyze citrus gene function. In this report, a rapid, universal, and highly efficient hairy root transformation system in citrus seeds was described. Only 15 days were required for the entire workflow and the system was applicable for various citrus genotypes, with a maximum transformation frequency of 96.1%. After optimization, the transformation frequency of Citrus sinensis, which shows the lowest transformation frequency of 52.3% among four citrus genotypes initially, was increased to 71.4% successfully. To test the applicability of the hairy roots transformation system for gene functional analysis of citrus genes, we evaluated the subcellular localization, gene overexpression and gene editing in transformed hairy roots. Compared with the traditional transient transformation system performed in tobacco leaves, the transgenic citrus hairy roots displayed a more clear and specific subcellular fluorescence localization. Transcript levels of genes were significantly increased in overexpressing transgenic citrus hairy roots as compared with wild-type (WT). Additionally, hairy root transformation system in citrus seeds was successful in obtaining transformants with knocked out targets, indicating that the Agrobacterium rhizogenes-mediated transformation enables the CRISPR/Cas9-mediated gene editing. In summary, we established a highly efficient genetic transformation technology with non-tissue-culture in citrus that can be used for functional analysis such as protein subcellular localization, gene overexpression and gene editing. Since the material used for genetic transformation are roots protruding out of citrus seeds, the process of planting seedlings prior to transformation of conventional tissue culture or non-tissue-culture was eliminated, and the experimental time was greatly reduced. We anticipate that this genetic transformation technology will be a valuable tool for routine research of citrus genes in the future.

Progress of clinical study on hypofractionated radiotherapy after breast-conserving surgery.

Whole-breast radiotherapy after breast-conserving surgery (BCS) can improve patient survival while reducing local tumor recurrence. Although standard breast radiotherapy can achieve good tumor control and cosmetic effects with low toxicity, the 5- to 7-week treatment time is relatively long for patients and can result in wasted medical resources. Therefore, there is a growing trend toward hypofractionated radiotherapy (HFRT), which accelerates partial-breast irradiation. Both short-course radiotherapy and conventional fractionated radiotherapy are safe and effective treatment modes, with similar survival and local tumor control effects as those of conventional radiotherapy (CRT), and adverse reactions can be tolerated. Compared with conventional fractionated radiotherapy, short-course radiotherapy saves medical resources and has a shorter total treatment time, reduced treatment costs, and an improved quality of life for patients.

[Quantitative estimation source of urban atmospheric CO2 by carbon isotope composition].

To effectively reduce urban carbon emissions and verify the effectiveness of currently project for urban carbon emission reduction, quantitative estimation sources of urban atmospheric CO2 correctly is necessary. Since little fractionation of carbon isotope exists in the transportation from pollution sources to the receptor, the carbon isotope composition can be used for source apportionment. In the present study, a method was established to quantitatively estimate the source of urban atmospheric CO2 by the carbon isotope composition. Both diurnal and height variations of concentrations of CO2 derived from biomass, vehicle exhaust and coal burning were further determined for atmospheric CO2 in Jiading district of Shanghai. Biomass-derived CO2 accounts for the largest portion of atmospheric CO2. The concentrations of CO2 derived from the coal burning are larger in the night-time (00:00, 04:00 and 20:00) than in the daytime (08:00, 12:00 and 16:00), and increase with the increase of height. Those derived from the vehicle exhaust decrease with the height increase. The diurnal and height variations of sources reflect the emission and transport characteristics of atmospheric CO2 in Jiading district of Shanghai.

[Size distributions of organic carbon (OC) and elemental carbon (EC) in Shanghai atmospheric particles].

Size distributions of organic carbon (OC), elemental carbon (EC) and secondary organic carbon (SOC) in atmospheric particles with size range from < 0.49, 0.49-0.95, 0.95-1.50, 1.50-3.00, 3.00-7.20, > 7.20 microm, collected in Jiading District, Shanghai were determined. For estimating size distribution of SOC in these atmospheric particles, a method of determining (OC/EC)(pri) in atmospheric particles with different sizes was discussed and developed, with which SOC was estimated. According to the correlation between OC and EC, main sources of the particles were also estimated roughly. The size distributions of OC and SOC showed a bi-modal with peaks in the particles with size of < 0.49 microm and > 3.0 microm, respectively. EC showed both of a bi-modal and tri-modal. Compared with OC, EC was preferably enriched in particles with size of < 0.49 microm. Mass concentrations of OC and EC in fine particles (< 3.00 microm) accounted for 59.8%-80.0% and 58.1%-82.4% of those in total suspended particles. OC and EC were preferably enriched in fine particles (< 3.00 microm). The concentrations of SOC in the particles with different sizes accounted for 15.7%-79.1% of OC in the particles with corresponding size. Concentrations of SOC in fine aerosols (< 3.00 microm) and coarse aerosols (> 3.00 microm) accounted for 41.4% and 43.5% of corresponding OC. Size distributions of OC, EC and SOC showed time-dependence. The correlation between OC and EC showed that the main contribution to atmospheric particles in Jiading District derived from light petrol vehicles exhaust.