Effects of Biostimulation-Bioaugmentation on Coastal Microbial Community in an in situ Mesocosm System

Globally,various types of pollution affect coastal waters as a result of human activities.Bioaugmentation and biostimulation are effective methods for treating water pollution.However,few studies have explored the response of coastal prokaryotic and eukaryotic communities to bioaugmentation and biostimulation.Here,a 28-day outdoor mesocosm experiment with two treatments(bioaugmentation-A and combined treatment of bioaugmentation and biostimulation-AS)and a control(untreated-C)were carried out.The experiment was conducted in Meishan Bay to explore the composition,dynamics,and co-occurrence patterns of prokaryotic and eukaryotic communities in response to the A and AS using 16S rRNA and 18S rRNA gene amplicon sequencing.After treatment,Gammaproteobacteria and Epsilonproteobacteria were significantly increased in group AS compared to group C,while Flavobacteriia and Saprospirae were significantly reduced.Dinoflagellata was significantly reduced in AS compared to C,while Chrysophyta was significantly reduced in both AS and A.Compared to C,the principal response curve analyses of the prokaryotic and eukaryotic communities both showed an increasing trend followed by a decreasing trend for AS.Furthermore,the trends of prokaryotic and eukaryotic communities in group A were similar to those in group AS compared with group C,but AS changed them more than A did.According to the species weight table on principal response curves,a significant increase was observed in beneficial bacteria in prokaryotic communities,such as Rhodobacterales and Oceanospirillales,along with a decrease in autotrophs in eukaryotic communities,such as Chrysophyta and Diatom.Topological properties of network analysis reveal that A and AS complicate the interactions between the prokaryotic and eukaryotic communities.Overall,these findings expand our understanding of the response pattern of the bioaugmentation and biostimulation on coastal prokaryotic and eukaryotic communities.

Study on the Retention Rate of Green Tea Catechins Components and Tea Polyphenol by Different Processing Technology

The aim of this research was to explore the effect of fixation,rolling,and drying processing technology on the retention rate of green tea catechins components and tea polyphenol.Different fixation processes(rotary drumfiring,microwave,steam-blasting),rolling process(weight of rolling,gently press rolling and traditional rolling),drying process(stove drying,roasting dehydration,baked fried drying) were adopted.The effect of different tea processing technology on the retention rate of catechins component and tea polyphenol was analyzed.It showed that the microwave fixation process,gently press rolling process,baked fried dry process were beneficial to keep high levels of EGCG,C,EGC,EC,ECG.

Genetic Transformation of Chinese Cabbage (Brassica rapa pekinensis) with Floral-dip Method

[ Objective] This study is aimed to transform gus gene into Chinese cabbage （Brassica rapa pekinensis） by floral-dip method. [ Method] The Chinese cabbage was transformed by floral-dip method for the first time. Then the cabbage seeds were harvested and screened by hygromycin, and the plants with resistance were confirmed by histochemical GUS assay and PCR detection. [ Result] The target gene was successfully integrated into the Chinese cabbage genome, and the transformation rate was 0.1%. [ Conclusion ] This study optimized the genetic transformation system for Chinese cabbage, and laid the foundation for improving the means of genetic transformation of Chinese cabbage.

Effect of Dietary Mulberry Leaves on Productive Performance,Egg Quality and Blood Biochemistry in Laying Hens

[Objective] The aim was to provide more information about the possible use of mulberry leaves in hen diets.[Method] Two hundred ninety two Hyline Gray laying hens, 48 wk old, were randomly allotted into four groups with 73 chickens per group. Each group was fed for 6 wk with diets that contained dried mulberry leaves at 0, 5, 10, or 15%. [ Result] The rate of egg production, egg weight, egg mass, feed intake, body weight, blood glucose level, total blood cholesterol, and triglyeride, and cholesterol of egg yolk in the dietary mulberry leaf groups were lower than the 0% mulberry leaves group. Compared with the 0% group, the feed required to produce 1 kg of eggs was 12.8% more in the 15% dietary mul- berry leaf group. Body weight and cholesterol of egg yolk decreased significantly in this group （P 〈0.05）. Blood glucose and triglyerides in the 15% dietary mulberry leaf group were significantly lower than in the 0 and 5% dietary mulberry leaf groups （P 〈0.05）. Yolk color in all treatments was higher than in the 0% group （P 〈0.05）, and Roche pigmentation score increased by 2 -2.9. One -deoxynojirimycin was detected from egg white in all treatments, and its content varied significantly between groups （ P 〈0.05）. E Conclusion] This study reflected the hypoglycemic and lipid -low- ering effects of mulberry leaves on productive performance and egg quality in laying hens. The suggested proportion of dietary mulberry leaves was 5% -10%.

Research Progress on Application of Ipomoea aquatica Floating Bed in Eutrophication Water

Research progresses on growth characteristics,application effect and optimization technology of I. aquatica floating bed were reviewed;its application problems in eutrophication water were discussed,and its application prospect was forecasted.

A Study on Physiological Character of Fresh Tea Leaves in Different Cold-Resistant Varieties

In order to reveal the cold resistant mechanism of tea plant,two varieties with different cold resistances were studied.The activities of catalase(CAT),peroxidase(POD),superoxide dismutase(SOD) and polyphenol oxidase(PPO) were investigated,and the contents of soluble protein,sugar and malondialdehyde(MDA) were determined in fresh tea leaves in the autumn.The results showed that the activities of POD and the content of soluble sugar were both much higher in the variety with strong cold resistance,and the content of soluble protein was associated with the cold resistance,while the cold resistance of the plant was not related with the activities of the CAT,SOD and PPO,and the content of MDA.It is considered that the activity of POD and contents of soluble sugar could be used as indicators to identify the cold resistance of tea varieties.

The transcriptional landscape of cultivated strawberry(Fragaria×ananassa)and its diploid ancestor(Fragaria vesca)during fruit development

Cultivated strawberry(Fragaria×ananassa)originated from four diploid ancestors:F.vesca,F.viridis,F.iinumae and F.nipponica.Among them,F.vesca is the dominant subgenome for cultivated strawberry.It is not well understood how differences in gene expression between diploid and octoploid strawberry contribute to differences during fruit development.In this study,we used comprehensive transcriptomic analyses of F.vesca and F.×ananassa to investigate gene expression at the different stages of fruit development.In total,we obtained 3508(turning stage)and 3958(red stage)differentially expressed genes with pairwise comparisons between diploid and octoploid.The genes involved in flavonoid biosynthesis were almost upregulated in the turning stages of octoploid,and we also discovered a ripe fruit-specific module associated with several flavonoid biosynthetic genes,including FveMYB10,FveMYB9/11,and Fve RAP,using weighted gene coexpression network analysis(WGCNA).Furthermore,we identified the species-specific regulated networks in the octoploid and diploid fruit.Notably,we found that the WAK and F-box genes were enriched in the octoploid and diploid fruits,respectively.This study elucidates new findings on flavonoid biosynthesis and fruit size of strawberry with important implications for future molecular breeding in cultivated strawberry.

The red/far-red light photoreceptor FvePhyB regulates tissue elongation and anthocyanin accumulation in woodland strawberry

Light is an important environmental signal that influences plant growth and development.Among the photoreceptors,phytochromes can sense red/far-red light to coordinate various biological processes.However,their functions in strawberry are not yet known.In this study,we identified an EMS mutant,named P8,in woodland strawberry(Fragaria vesca)that showed greatly increased plant height and reduced anthocyanin content.Mapping-by-sequencing revealed that the causal mutation in FvePhyB leads to premature termination of translation.The light treatment assay revealed that FvePhyB is a bona fide red/far-red light photoreceptor,as it specifically inhibits hypocotyl length under red light.Transcriptome analysis showed that the FvePhyB mutation affects the expression levels of genes involved in hormone synthesis and signaling and anthocyanin biosynthesis in petioles and fruits.The srl mutant with a longer internode is caused by a mutation in the DELLA gene FveRGA1(Repressor of GA1)in the gibberellin pathway.We found that the P8 srl double mutant has much longer internodes than srl,suggesting a synergistic role of FvePhyB and FveRGA1 in this process.Taken together,these results demonstrate the important role of FvePhyB in regulating plant architecture and anthocyanin content in woodland strawberry.

A Simplified Rice DNA Extraction Protocol for PCR Analysis

A simple protocol was established for DNA extraction using etiolated rice seedlings, whereby rice DNA was directly extracted in 0.5 mol/L NaOH solution in a single eppendorf tube. Results of comparative PCR analyses and electrophoresis showed that the DNA extracted using this method was as good and useful as that using standard CTAB method.

Research on Intelligent Monitoring System Based on Raspberry Pi 3

In order to monitor environmental parameters for food storage more effectively in real time,an intelligent monitoring system was designed and implemented based on Raspberry Pi.Based on the Raspberry Pi 3 Model B,the system connects environmental sensors such as temperature,humidity,light intensity and CO2 concentration.It can be used to monitor environmental parameters such as air temperature and humidity,light intensity and CO2 concentration during food storage.Based on Raspberry Pi 3,this study successfully built an intelligent monitoring system,and developed web version for computer terminal and app(iOS version and Android version)for mobile terminal.The data about the environmental factors during food storage were obtained.The system is reliable,simple and practical in operation,and highly expandable,laying a foundation for the next step of research on food safety storage environmental parameters and intelligent control.

Two ABCI family transporters,OsABCI15 and OsABCI16,are involved in grain-filling in rice

Seed development is critical for plant reproduction and crop yield,with panicle seed-setting rate,grain-filling,and grain weight being key seed characteristics for yield improvement.However,few genes are known to regulate grain filling.Here,we identify two adenosine triphosphate(ATP)-binding cassette(ABC)I-type transporter genes,OsABCI15 and OsABCI16,involved in rice grain-filling.Both genes are highly expressed in developing seeds,and their proteins are localized to the plasma membrane and cytosol.Interestingly,knockout of OsABCI15 and OsABCI16 results in a significant reduction in seed-setting rate,caused predominantly by the severe empty pericarp phenotype,which differs from the previously reported low seed-setting phenotype resulting from failed pollination.Further analysis indicates that OsABCI15 and OsABCI16 participate in ion homeostasis and likely export ions between filial tissues and maternal tissues during grain filling.Importantly,overexpression of OsABCI15 and OsABCI16 enhances the seed-setting rate and grain yield in transgenic plants and decreases ion accumulation in brown rice.Moreover,the OsABCI15/16 orthologues in maize exhibit a similar role in kernel development,as demonstrated by their disruption in transgenic maize.Therefore,ourfindings reveal the important roles of two ABC transporters in cereal grain filling,highlighting their value in crop yield improvement.

Comparative Proteomic Analysis of Arabidopsis Mature Pollen and Germinated Pollen

Proteomic analysis was applied to generating the map of Arabidopsis mature pollen proteins and analyzing the differentially expressed proteins that are potentially involved in the regulation of Arabidopsis pollen germination. By applying 2-D electrophoresis and silver staining, we resolved 499 and 494 protein spots from protein samples extracted from pollen grains and pollen tubes, respectively. Using the matrix-assisted laser desorption ionization time-of-flight mass spectrometry method, we identified 189 distinct proteins from 213 protein spots expressed in mature pollen or pollen tubes, and 75 new identified proteins that had not been reported before in research into the Arabidopsis pollen proteome. Comparative analysis revealed that 40 protein spots exhibit reproducible significant changes between mature pollen and pollen tubes. And 21 proteins from 17 downregulated and six upregulated protein spots were identified. Functional category analysis indicated that these differentially expressed proteins mainly involved in signaling, cellular structure, transport, defense/stress responses, transcription, metabolism, and energy production. The patterns of changes at protein level suggested the important roles for energy metabolism-related proteins in pollen tube growth, accompanied by the activation of the stress response pathway and modifications to the cell wall.

Quantitative detection of Cymbidium mosaic virus by real time PCR

The technique of SYBR Green-based quantitative real-time reverse transcription polymerase chain reaction(real-time RT-PCR)was applied to quantitative detect a 764 bp nucleotide sequence containing total coat protein(cp)gene of Cymbidium mosaic virus(CyMV).The plasmid containing the target sequence was constructed to prepare the standard curve and detect the sensitivity.The standard curve was drawn based on the linear relationship between the logarithm(base 10)of the quantity of target sequence and cycle threshold[C(T)].While the concentration of plasmid DNA falling within the range of 2.6×10^(7)to 2.6×10^(2)copies per tube established a regression equation,y=-0.3583x+10.32,and related coefficient:r^(2)=0.995.The real-time RT-PCR assay for CyMV had a minimum detectable quantity of two copies per tube.The naturally infected samples of Phalaenopsis sp.and the artificially inoculated samples of Arachnis sp.with trace CyMV were quantitatively detected using this method.CyMVin the positive samples of Phalaenopsis sp.and Arachnis sp.was confirmed by DNA sequencing and cp gene homeology blast.The results showed that CyMV extracted from the leaves of orchid in Hangzhou,Zhejiang Province,China,could be derived from Kunming city(KM),Yunnan Province,China.This method characterized by high sensitivity,specificity,and precision is suitable for early diagnosis and quantitative detection of CyMV.

Status of water environment pollution in the Xixi Wetland and its ecological treatment countermeasures

In this paper,the water quality of the Xixi Wetland was evaluated and the characteristics of water pollution were described according to the survey data.Based on the status of water quality and its functional requirements as an urban wetland,biological-ecological countermeasures were suggested.The experimental use of ecological technologies,such as artificial wetlands,ecological aquiculture and artificial floating island,were done in several fish ponds in the Xixi Wetland.Water monitoring results show that the quality of the treated water has improved significantly and the measures to purify the eutrophic water in the wetland have been effective.

Cadmium induced a non-coding RNA microRNA535 mediates Cd accumulation in rice

Identifying key regulators related to cadmium(Cd)tolerance and accumulation is the main factor for genetic engineering to improve plants for bioremediation and ensure crop food safety.MicroRNAs(miRNAs),as fine-tuning regulators of genes,participate in various abiotic stress processes.MiR535 is an ancient conserved non-coding small RNA in land plants,positively responding to Cd stress.We investigated the effects of knocking out(mir535)and overexpressing miR535(mir535 and OE535)under Cd stress in rice plants in this study.The mir535 plants showed better Cd tolerance than wild type(WT),whereas the OE535 showed the opposite effect.Cd accumulated approximately 71.9%and 127%in the roots of mir535 and OE535 plants,respectively,compared to WT,after exposure to 2μmol/L Cd.In brown rice,the total Cd accumulation of OE535 and mir535 was about 78%greater and 35%lower than WT.When growing in 2 mg/kg Cd of soil,the Cd concentration was significantly lower in mir535 and higher in OE535 than in the WT;afterward,we further revealed the most possible target gene SQUAMOSA promoter binding-like transcription factor 7(SPL7)and it negatively regulates Nramp5 expression,which in turn regulates Cd metabolism.Therefore,the CRISPR/Cas9 technology may be a valuable strategy for creating new rice varieties to ensure food safety.

A zinc finger protein, interacted with cyclophilin,affects root development via IAA pathway in rice

The plant hormone auxin plays a crucial role in lateral root development. To better understand the rnolecular mechanisms underlying lateral root formation, an auxin-responsive gene OsCYP2 （Oso2g0121300） was characterized from rice. Compared to the wild type, OsCYP2-RNAi （RNA interference） lines exhibited distinctive defects in lateral root development. Yeast two-hybrid and glutathione S-transferase puIl-down results confirmed that OsCYP2 interacted with a C2HC-type zinc finger protein （OsZFP, O501g0252900） which is located in the rice nucleus. T2OsZFP-RNAi lines had significantly fewer lateral roots than did wild-type plants, which suggests a role for OsCYP2 and OsZFP in regulating lateral root development.Quantitative real-time polymerase chain reaction showed that the expression of certain Aux/IAA （auxin/indole-3- acetic acid） genes was altered in OsCYP2- and OsZFP-RNAi lines in response to IAA. These findings imply that OsCYP2 and OsZFP participate in IAA signal pathways controlling lateral root development. More importantly, OslAA11 showed functional redundancy not only in OsCYP2-RNAi lines but also in OsZFP-RNAi lines, which provides important clues for the elucidation of mechanisms controlling lateral root development in response to auxin.

Suitable soil moisture contents for water use efficiency and saponins accumulation in Panax notoginseng

Objective:The moisture content in the soil directly affects the yield and quality of Panax notoginseng,especially at the age of three years old.However,the suitable moisture for the growth of P.notoginseng is unknown.In this study,the effects of different soil moisture on the growth of P.notoginseng were studied.Methods:Four different water treatments(0.45 field capacity(FC),0.60 FC, 0.70 FC, and 0.85 FC) were set up in Shilin County,Yunnan Province,China.The water consumption and daily dynamic of water consumption were determined daily(from April 21 to October 18,2012),and the daily dynamic of water consumption under different weather conditions(sunny and rainy) was determined.The transpiration coefficient and water use efficiency were calculated through dry matter accumulation and total water consumption.Accumulation of saponins of roots of P.notoginseng were analyzed by HPLC after treated,and the soil moisture content suitable for the growth of P.notoginseng was estimated by regression fitting of the active ingredient accumulation and the soil moisture content.Results:The water consumption of 0.85 FC,0.70 FC,0.60 FC and 0.45 FC were 2.89,3.68,3.37 and 2.73 kg/plant per day,respectively.The water consumption of P.notoginseng from June to August was greater than other months.The daily dynamic of water consumption on sunny days and sunny days after rain showed a "double peak" feature,and it showed a "single peak" feature on rainy days.The water uses efficiency(WUE) of 0.85 FC,0.70 FC,0.60 FC and 0.45 FC were 2.51,3.32,4.59,3.39 gDW/kg H_(2)O,respectively.The increase of soil moistu re content would reduce the WUE of P.notoginse ng.With the increase of soil water content,the content of notoginsenoside R_(1) and ginsenoside Rg_(1) did not change significantly,while the content of ginsenoside Rb_(1) and Rd showed a decreasing trend.Conclusion:Soil moisture content significantly affected the water consumption of P.notoginseng,and when it was 56.4% of the maximum water holding capacity in the field,the sum of the four saponins of 100 strains of P.notoginseng was the highest.