It is important to investigate the mechanism of growth and development of edible fungi for genetic breeding and cultivation technique research. Based on the total expressed proteins of biological tissues and cells, a protein expression profile can be established by using proteomics technology, which can not only provide a material basis for the law of life, but also provide a theoretical basis and solution for biological genetic breeding.
In this study, in order to analyze the protein expression changes at different developmental phases and explore the law of growth and development of Pleurotus pulmonarius, we applied an integrated proteomic approach to comprehensively investigate the proteome change of P. pulmonarius at different developmental phases, including mycelium phase, primordium phase, coral-like phase, and fruit body phase, by using high-sensitive-mass spectrometer.
The results showed that, compared with the initial mycelium growth phase, a total of 885 differentially expressed proteins were isolated from the other three growth phases, with 376, 642 and 692 differentially expressed proteins from the primordium phase, coral-like phase and fruit body phase, respectively. The specifically expressed proteins were 11, 23, 136 and 181 respectively in the four different developmental phases. Gene ontology annotation showed that 26%, 20%, 15% and 11% of the differentially expressed proteins were related with carbohydrate synthesis, reproductive structure development, organ synthesis and protein metabolism. Moreover, the analysis of metabolic pathway showed that the differentially expressed proteins mainly involved in the pathway of tricarboxylic acid cycle and glycolysis/gluconeogenesis which may possibly play an important role in the growth and development of P. pulmonarius. The heat shock protein 70 (HSP70), belonging to a molecular chaperone, showed significant differences in mycelium phase, primordium phase, corallike phase, and fruit body phase, indicating that it is critical to regulate the growth of P. pulmonarius.
In conclusion, there are differentially expressed proteins and differential signaling pathways at different developmental phases of P. pulmonarius. These differentially expressed proteins play a very important role in the organic development, fruiting body morphogenesis and resistance to adverse environmental conditions. This study lays a theoretical basis for the research of breeding and cultivation techniques of P. pulmonarius.

Abscisic acid (ABA) is a vital hormone that regulates stomatal closure and responds to stress conditions in plants. Therefore, ABA signal pathway involves numerous physiological processes in plants, such as seed dormancy, germination, development and ripening. Among the families of ABA receptor genes, ABA-insensitive (ABI) is a major transcription factor that regulates ABA signal pathway. To date, little information is known about the ABI gene of buckwheat.
To characterize ABI gene sequence polymorphism and genetic relationship of genus Fagopyrum, the budlets of 30 germplasm resources from seven wild species including F. esculentum, F. tataricum, F. zuogongense, F. pilus, F. megaspartanium, F. cymosum and F. urophyllum were collected for DNA extraction. The ABI UniGene sequence of buckwheat was isolated from the previously performed transcriptome sequencing data (accession number: SRS1350375). Specific primers of ABI gene were designed based on the high conservative sequence for polymerase chain reaction (PCR) amplification, and the amplified fragments from 30 germplasm resources were processed for gene sequencing. Then the gene sequence analysis was performed on nucleotide
BLAST of NCBI database (http://blast.ncbi.nlm.nih.gov/Blast.cgi), and the genetic variance was analyzed by DNAsp5, and the analysis of ClustalW multiple sequence alignment, genetic distance and hierarchy were clustered by MEGA 5.05.
The results showed that 20.28% of polymorphic sites were found in ABI gene fragments among 30 germplasm resources of the seven wild species of genus Fagopyrum, whereas only 1.86% of polymorphic sites were found among 14 germplasm resources of F. tataricum and 1.81% of polymorphic sites were found among five germplasm resources of F. esculentum, indicating that ABI gene sequence is highly conserved in the intraspecies of genus Fagopyrum. Genetic distance analysis showed that both the values of interspecific genetic distance and interspecific net genetic distance between F. zuogongense and F. pilus were the largest, whereas those between F. cymosum and F. urophyllum were the least based on the ABI gene sequence analysis. Hierarchical cluster analysis indicated that the phylogenetic relationship of F. tataricum was close to F. megaspartanium, F. cymosum and F. urophyllum, then further grouped with F. pilus, while the F. esculentum was close to F. zuogongense based on the ABI gene sequence analysis. The F. tataricum and F. zuogongense collected from different regions were respectively clustered, suggesting that the mutation of ABI gene might be influenced by the growth environment during plant evolution.
Inconclusion, ABI gene sequence is highly conserved in the intraspecies of genus Fagopyrum; nevertheless, different growth environments caused distinctive mutations of ABI gene. Based on the gene sequence analysis, a close relationship was found between F. esculentum and F. zuogongense. In addition, close relationships were also found among F. tataricum, F. megaspartanium, F. cymosum and F.urophyllum.The above results provide a theoretical basis for ABI gene research and evolutionary relationship of genus Fagopyrum, but the exact evolution among different species of buckwheat still needs further research.

During development process of the Zhoushan archipelago, the original old-growth broadleaved forests on the islands have been severely destroyed due to intensive human activity. Neolitsea sericea (Lauraceae), distributed on a few islands of the Zhoushan archipelago, is facing the danger of extinction due to rapid degradation and destruction of original habitats. So far, plenty of studies on N. sericea have been focused on its population genetic diversity and genetic structure. Neolitsea sericea is well adaptable to the environment, thus can survive in the ravine on the islands. The strong tolerance of N.sericea to drought, wind, salt and barren soil also makes it optimal species for afforestation. However, few studies on drought tolerance of N. sericea have been reported yet.
Plant response to drought stress at a molecular level is a complex biological process, involving consideration of the stress effects and regulation events. Thus, it is of great importance to analyze the underlying mechanism systematically at the physiological level.
In this study, one-year-old seedlings of N. sericea were selected as test materials. To obtain a more complete physiological mechanism in responses to drought stress, N. sericea was exposed under the conditions of four different relative water contents (normal water supply, light drought, moderate drought and severe drought) in soil for 45 days. The relative water content in soil of normal water supply, light drought, moderate drought, and severe drought was controlled in 75%-80%, 55%-60%, 40%-45% and 30%-35% of field capacity, respectively. All plants were rehydrated after drought treatments by normal water supply for 10 d. The relative water content in soil was controlled by pot- weighing method through the experiment. The dynamic changes of the physiological and photosynthetic traits in leaves were measured every 15 days of stress treatments and at the end of rehydration. The traits measured in this study include net photosynthetic rate (Pn), stomatal conductance (Gs), water use efficiency (WUE), intercellular CO₂ concentration (Ci), transpiration rate (Tr), total content of chlorophyll (Chl), superoxide dismutase (SOD) and peroxidase (POD) activities, proline content, malondialdehyde (MDA) content, soluble sugar content and relative conductivity (Rc).
The results showed that the contents of Chl, Pn, Tr and Gs decreased constantly and significantly during severe drought, and the content of Ci decreased in 0-30 d but enhanced in 30-45 d during the severe drought, which indicated that the decrease of Pn was caused by stomatal limitation during the 0-30 d of severe drought, and by non-stomatal limitation during 30-45 d of severe drought. The continuous inhibition of Tr after rehydration from severe drought suggested the afunction of stomatal regulation. The drought stress increased the overall WUE of the leaves, although the WUE dropped significantly in 30-45 d of severe drought. The trend of MDA content, SOD and POD activities in N. sericea first increased and then decreased during the drought treatments, indicating the production and clearance of MDA had reached a dynamic balance. The significant increase of Rc indicated the elevation of cell membrane permeability. The overexpression of soluble sugar and proline in leaves during 30-45 d of drought treatment resulted in the reduction of Rc, which relieved the osmotic pressure in N. sericea. However, Rc staying at a high level after rehydration from severe drought indicated the damage of cell membrane.
In conclusion, N. sericea can conduct the instant stomatal regulation, eliminate active oxygen and regulate the osmotic pressure effectively, decrease the Tr and MDA content, increase the cell membrane permeability and enhance the WUE of leaves, to reduce the damage under the drought stress. However, sev

As is well-known, the iodine deficiency disorders (IDD) have tremendous adverse effects on the growth and development of human beings. The universal salt iodization (USI) has been introduced for the control and elimination of IDD in many countries. However, excessive iodine intake caused by USI may lead to new diseases such as hyperthyreosis, high level iodine goiter and thyroid cancer. Meanwhile, other investigations indicated that organic iodine is much safer than inorganic iodine added in salt.
A new strategy has been confirmed by pot and hydroponic experiments to cultivate the iodine- rich crops through biofortification for iodine supplement. This study aims to testify the feasibility of cultivating iodine-rich crops in a large scale under natural conditions, thus exploring a potential way for the prevention and elimination of the IDD.
Randomized block experiments were conducted in the farmland in Sha’an Village of Bihu Basin, which located in Lishui City of Zhejiang south mountain area, to explore the biofortification effect of seaweed iodine fertilizer on iodine contents of rice, vegetables and fruits in the field. The seaweed iodine fertilizer was prepared by mixing the smashed kelp with diatomite according to patent formula. The treatments of exogenous iodine were as follows：0, 0.375, 0.750, 1.500, 3.000, and 4.500 kg/hm². Spectrophotometric method was used to detect the iodine contents of crops and soil samples, and one-way analysis of variance was applied to analyze the difference of iodine contents among all treatments.
The results suggested that the iodine content in the edible parts of all the tested crops were significantly enhanced (P＜0.05) after biofortification by the seaweed iodine fertilizer, including rice, loofah, cucumber, tomato, eggplant, watermelon, edamame, cowpea, etc. The iodine content of rice grain on a dry mass basis increased from 0.019 mg/kg to 0.403 mg/kg after iodine biofortification, while the iodine content of the vegetables and fruits on a fresh mass basis may be enhanced from 0.01-0.05 mg/kg to 0.130 mg/kg after iodine application. Besides, the iodine translocation factors from the roots to the edible parts of cowpea and edamame were relatively higher than the others. A daily consumption of 0.38 kg fresh iodine-rich vegetables or fruits and 0.40 kg iodine-biofortified rice can offer 150 μg/d iodine for adults, which can meet the daily iodine intake recommended by the World Health Organization (WHO).
In conclusion, the iodine content varied significantly in different parts of rice, and increased gradually from roots to straws and grains. Low concentration (0.375 kg/hm²) of exogenous iodine had little effect on the iodine content of the edible parts of crops, while high concentration (≥3.000 kg/hm²) of exogenous iodine could reduce the iodine content of the edible parts of crops. Upon overall consideration of the costs and benefits for the plantation, 1.500 kg/hm² should be an optimal exogenous iodine dosage for the cultivation of iodine-rich crops in the field.

Eggplant (Solanum melongena L.) is one of nutritious vegetables welcomed by consumers. However, the diseases occurred on eggplants, particularly soil-borne diseases became increasingly severe, which hampered sustainable development of eggplant production. Based on our investigation, eggplant rotation is able to reduce the occurrence of diseases significantly, but its mechanism has not been explained clearly.
In this study, we collected soil samples from different fields cultivated by eggplant continuous cropping and rotation to analyze their nutrients, enzymatic activities and microbial community structure using high throughput sequencing technique.
The results showed that the contents of available P, Ca, Mg and Mn decreased significantly, and the activities of polyphenol oxidase (PPO), urease (UE), alkaline protease (ALPT) decreased; the number of bacterial operational taxonomic units (OTUs) reduced but fungal OTUs increased, and the bacterial and fungal diversity indexes decreased and the dominance index increased in fields with four-year continuous cropping (eggplant-eggplant-eggplant-eggplant, EEEE), as compared with different eggplant rotation fields. However, there were significant differences in the nutrients, enzymatic activities and microbial community structure from different soil samples between the four-year eggplant rotation fields [eggplant-rice-rice-eggplant, ERRE or eggplant-green bean (the early half of the year)+bottle gourd (the latter half of the year)-green bean (the early half of the year)+bottle gourd (the latter half of the year)-eggplant, EGBE] as well. Compared with the EGBE, the pH and the contents of total P, available P, exchangeable Ca, Mg and Mn increased significantly, but the salinity and available boron (B) reduced significantly; the activities of peroxidase, acid protease, neutral protease decreased significantly but the catalase, UE and ALPT increased significantly; the number of bacterial OTUs increased significantly but fungal OTUs decreased; the bacterial and fungal diversity indexes increased and the dominance indexes decreased in ERRE.
In sum, ERRE is a good and worthy way for rotation by comprehensively considering soil pH, salinity, nutrients, enzymatic activities and microbial community structure in this study. The above results lay a theoretical foundation for choosing suitable cultivation system of eggplants in mountain regions.

The colored calla lily (Zantedeschia hybrida), as the “flower star of the 21st century”, is becoming more and more popular for its special morphology and high ornamental value. But at present, the varieties of colored calla lily in the market are imported from foreign countries like the Netherlands, New Zealand, Japan, etc. Without independent intellectual property rights, the industry of colored calla lily has been seriously restricted in China; therefore, breeding new
varietyies of colored calla lily with excellent ornamental traits, resistance to disease and strong growth vigor is of great significance to our country. Polyploids of ornamental plants are usually characterized by their superior growth vigor, flower size, stem sturdiness, and better resistance as compared with diploids. Two tetraploid variations of colored calla lily varieties “Tanswan” and “Liberty” were successfully established in the previous study; however, whether the tetraploid varaitions have more advantages in growth performance, morphological characteristics and resistance to stress than the diploids are still unclear.
In this study, the tissue culture plantlets of the diploid and tetraploid of colored calla lily were transplanted and managed at the same environmental condition and cultivation measure. Their ploidy level, survival rate, morphological characteristics (including plant height, stem diameter, maximum leaf length, maximum leaf width, leaf- shape index, leaf color, and uniformity), dry matter in bulb, resistance to stress and dormancy were investigated and analyzed.
The results showed that the chromosome number of two tetraploid variations was 64 after two months of transplanted into the field, suggesting the ploidy of the tetraploid variations are stable. The survival rate of colored calla lily cultured in the medium with more peat moss [V(peat moss):V(pelite)=6:4] was higher than that in the medium of V(peat moss):V(pelite)=7:3, which suggests the permeability of the medium is important to the growth of the colored calla lily. Compared with the diploid plants, the tetraploid plants of colored calla lily grew faster and stronger, had higher survival rate, thicker/ darker green leaves, earlier germination time and shorter dormant period. Furthermore, the tetraploid plants had higher survival rate with an increase of 23.59% and 26.74% respectively in “Tanswan” and “Liberty” than the diploids under the low temperature stress, indicating the tetraploids of colored calla lily have higher resistance to low temperature. Under the same environmental condition, the difference in fresh mass and dry mass of the bulbs was quite significant at different ploidy levels. The tetraploids had higher ratio of dry to fresh mass than diploids, which suggests that the tetraploids have stronger ability to accumulate dry matter than the diploids. By comparing and analyzing the content of free proline, activities of superoxide dismutase (SOD) and peroxidase (POD), we found the tetraploids of colored calla lily had higher resistance to low temperature than the diploids; meanwhile, the resistance to low temperature stress of colored calla lily was negative correlation with the changes of free proline content, and the varieties with higher resistance to low temperature
stress had a stronger antioxidation system.
In conclusion, the tetraploids of colored calla lily have significant advantages in morphological characteristics, dry matter accumulation in bulb, resistance to stress and dormancy characteristics; therefore, the tetraploid of colored calla lily has great potential to be applied the production in the future.

Polyphenol oxidase (PPO, EC 1.10.3.1) has great significance to the growth and development of Camellia sinensis, for instance it participates in adjusting and maintaining the normal physiology and metabolism, plays an essential role in the course of the quality formation of tea products, and contributes to the oxidation reaction of tea polyphenols. The oxidation products, such as quinines, theaflavins and thearubigins, have a remarkable effect on the quality of black tea. The existing researches of C. sinensis PPO involve in various varieties and multiple aspects, including separation and purification, enzymatic property analysis, genetic structure, etc. Nevertheless, there is no research on C. sinensis var. assamica cv. Mengku, even though it carries relative original genetic information and has a great suitability for black tea manufacture.
In this study, the fresh leaves of Camellia sinensis var. assamica cv. Mengku were used as materials. The crude polyphenol oxidase (PPO) from tea leaves was extracted by using homogenate extraction technology and purified through (NH₄)₂SO₄ precipitation. The characteristics including optimum reaction pH, optimum reaction temperature, substrate specificity, Michaelis-Menten constants and the effect of inhibitors and metal ions on its activity were studied. It aims to lay the foundation for regulating PPO activity and illuminating the suitability for black tea manufacture of C. sinensis var. assamica.
The results showed that the optimum pH of crude PPO was 4.4, and its optimum temperature was 37 ℃. It had the catalytic activity on catechol and pyrogallic acid, but not on hydroquinone and resorcinol. When pyrogallic acid and catechol as its substrate, their K_m values were 4.2 and 36.5 mmol/L, respectively, and their V_max values were 2 000 and 5 000 U/mL, respectively, indicating that the catalytic efficiency of pyrogallic acid was better. The inhibition of EDTA-2Na, sodium sulfite, ascorbic acid and L-cysteine on crude PPO was enhanced with the increasing concentration, and sodium sulfite had the strongest inhibitory effect on it, while EDTA-2Na showed the lowest inhibitory rate, with 3.64% in 1.00 mmol/L. In this experimental condition, citric acid showed no inhibitory effect on crude PPO, on the contrary its activity was slightly increased (P＞0.05). Na⁺ had inhibition action on the crude PPO, whereas both K⁺ and Mg²⁺ showed a promotion effect on the crude PPO. Cu²⁺ of 10 mmol/L increased the enzymatic activity of crude PPO to 172.85%, which shows that Cu²⁺ has a strong promotion effect on the crude PPO.
In conclusion, the relative lower optimal pH and optimum reaction temperature of the crude PPO are advantageous to the formation of main quality components in black tea, particularly the formation of theaflavins, which may account for that C. sinensis var. assamica cv. Mengku is fit to manufacture black tea. The characteristics of the crude PPO, which was extracted and purified from C. sinensis var. assamica cv. Mengku, show visible differences with other varieties. This may be caused by discrepancy of protein structure, and lead to suitability for manufacture of tea varieties.

Rapeseed is an oil crop with quite large growing areas in China, and also one of the main oil crops in the world. As one of the most serious diseases in rapeseed, Sclerotinia disease, caused by the fungal pathogen Sclerotinia sclerotiorum, can result in significant yield loss. At present, no rape cultivars are available that are resistant to S. sclerotiorum. Using fungicides is the main measure to control rapeseed Sclerotinia disease in China. Carbendazim, as a common fungicide, has been frequently applied to control rapeseed Sclerotinia disease for long. Increasing evidence of fungicide resistance in
populations of S. sclerotiorum has been found. Therefore, new fungicides are constantly being requested to control rapeseed Sclerotinia disease. Acetolactate synthase (ALS) is the enzyme that catalyzes the first step in the branched-chain amino acid biosynthesis pathway in plants and microbes, and is the target of some herbicides. Recently, some studies have reported the development of new antibiotics to control bacterial pathogens or new fungicides to control fungal pathogens of human beings with ALS as the action target. However, there are few studies on developing new fungicides to control fungal
pathogens of plants with ALS as the action target.
In the present study, seven potential ALS inhibitors (chlorimuron-ethyl, bensulfuron-methyl, chlorsulfuron, sulfometuronmethyl, imazethapyr, imazaquin and sulfathiazole) were used to determine the effectiveness of controlling rapeseed Sclerotinia disease. Chlorimuron-ethyl, bensulfuron-methyl, chlorsulfuron and sulfometuron-methyl were chosen as the representatives of sulfonylurea-type ALS inhibitors. Imazethapyr and imazaquin were the representatives of imidazolinone-type ALS inhibitors, and sulfathiazole was the representative of sulfathiazole-type ALS inhibitors.
The results indicated that the activity of ALS in S. sclerotiorum could not been inhibited by 1.0 mg/L imazethapyr. In contrast, the treatment with 1.0 mg/L chlorimuron-ethyl, bensulfuron-methyl, chlorsulfuron, sulfometuron-methyl, imazaquin and sulfathiazole showed more or less inhibitory effects on the ALS activity of S. sclerotiorum. Among them, imazaquin had the strongest inhibitory effect on ALS, but sulfometuron-methyl showed the weakest inhibitory effect. Imazaquin and sulfathiazole could effectively suppress the growth of S. sclerotiorum colony on potato dextrose agar plates at the concentration of 0.1 mg/L.
Chlorimuron-ethyl, bensulfuron-methyl and chlorsulfuron had inhibitory effect at the concentration of 1.0 mg/L. However, sulfometuron-methyl and imazethapyr showed weak inhibitory effect on the growth of S. sclerotiorum colony even at the concentration of 50 mg/L. The half-maximal effective concentrations (EC50) of imazaquin, sulfathiazole, chlorimuron-ethyl, bensulfuron-methyl and chlorsulfuron were determined to be less than 10 mg/L, but EC50 values of sulfometuron-methyl and imazethapyr were determined to be more than 50 mg/L. All the seven potential ALS inhibitors showed inhibitory effects on the activity of ALS in rapeseed leaves at the concentration of 1.0 mg/L. The inhibitory effects of chlorimuron-ethyl, bensulfuronmethyl, chlorsulfuron, imazaquin and sulfometuron-methyl were much stronger than those of imazethapyr and sulfathiazole. The inhibitory effect of sulfathiazole was the weakest among the seven inhibitors. Significant damaging influences were observed on rapeseed seedlings by using chlorimuron-ethyl, bensulfuron-methyl, chlorsulfuron, imazaquin, sulfometuron-methyl and imazethapyr at the concentration of 1.0 mg/L, such as decreasing growth rate, dwarfing, reducing leaf size and leaf yellowing, but no significant influences were observed by treating rapeseed seedlings with sulfathiazole. The use of sulfathiazole could exert remarkable inhibitory effects against ALS activity and colony growth of

The cigarette beetle, Lasioderma serricorne (Fabricius) (Coleoptera: Anobiidae), is a destructive stored pest distributed worldwide. Females typically oviposit in the stored materials, and the developing larvae tunnel through the stored food products on which they feed, resulting in tremendous damages to stored grains and seeds, tobacco, pet food, and dried flowers. Currently, excessive use of fumigation with insecticides has developed resistance to commonly insecticides in this insect, leading to environmental risk and its resurgence.
Controlled atmosphere (CA) treatments using low oxygen (O₂) and high carbon dioxide (CO₂) have been used commercially to control stored pests. The application of CA is a safe alternative way to chemical pesticides, and has been used to control the cigarette beetles. Our earlier studies showed that L. serricorne displayed a great capacity to withstand CO₂ stress. The activities of glutathione S-transferases (GSTs) in L. serricorne were significantly increased after exposed to CO₂ enriched atmosphere, implying that GSTs might be critical for tolerating to CO₂ stress. However, little is known about the molecular mechanism of GSTs in response to CO₂ exposure in L. serricorne.
In this study, three full-length cDNAs of GSTs in L. serricorne were cloned by reverse transcription-polymerase chain reaction (RT- PCR). The deduced amino acid sequence analysis of the three genes was performed by bioinformatics methods. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to establish expression profiles for those genes from different developmental stages and in response to CO₂ exposure. According to the predicted GST sequences from the transcriptome datesets of L. serricorne, the full-length cDNAs of three novel genes were obtained and named LsGSTt1, LsGSTd1 and LsGSTs1 (GenBank accession numbers: KY549655, KY549656, KY549657), respectively. The LsGSTt1 cDNA contained an open reading frame (ORF) of 564 nucleotides, encoding a polypeptide of 187 amino acids with an estimated molecular mass of 22.4 kDa and an isoelectric point (pI) of 8.54. The LsGSTd1 cDNA contained an ORF of 651 nucleotides, encoding a polypeptide of 216 amino acids with an estimated molecular mass of 24.5 kDa and a pI of 5.95. The LsGSTs1 cDNA contained an ORF of 696 nucleotides, encoding a polypeptide of 231 amino acids with an estimated molecular mass of 26.2 kDa and a pI of 5.06. Homology analysis indicated that the predicted amino acid sequences of the three genes had typical features of GSTs, including an N-terminal domain and a C-terminal domain. Phylogenetic analysis revealed that the three genes belonged to three different cytosolic classes, including Theta (LsGSTt1), Delta (LsGSTd1) and Sigma (LsGSTs1). Temporal expression profile revealed that all the three GSTs genes were constitutively expressed in the testing stages, exhibited similar developmental expression patterns, with the highest expression level measured in the early larval stage. Compared with the control, the mRNA levels of LsGSTd1 did not change significantly following exposure to CO₂ stress. However, when exposed to 50% lethal concentration (LC50) of CO₂, the expression levels of LsGSTt1 and LsGSTs1 were significantly increased.
In conclusion, the results suggest that LsGSTt1 and LsGSTs1 in L. serricorne might play important roles in defense responses challenged by CO₂ stress. This study provides the basis for clarifying the response mechanism of insects to CO₂ stress.

Pomacea canaliculata is an introduced aquatic animal which has transformed into a malignant aquatic animal of paddy and other aquatic plants in China. In addition, the snail also acts as an intermediate host of Angiostrongylus cantonensis, and the main causative agent of human eosinophilic encephalitis. Moreover, the snail consumes dissolved oxygen in freshwater, causing destruction of the aquatic environment. As a result, P. canaliculata was listed as an invasive species in China by the Chinese State Environmental Protection Administration in 2003. It is necessary to prevent the species from spreading further and expanding its population. Pueraria peduncularis Grah. is a traditional Chinese medicine that belongs to the Pueraria family within the Leguminosae, and distributes widely across southwestern China. In terms of agricultural activity, the herb is biologically active against a variety of agricultural pests, and has insecticidal, bactericidal, and slug-killing activities. In our previous study, we found that P. peduncularis extracts had extremely strong toxicity against the apple snail and further identified the active compound as pedunsaponin, which has LC₅₀ value of 3.89 mg/L and great molluscicidal potential.
To explore the effects of pedunsaponin on the respiratory metabolism of P. canaliculata, the oxygen consumption rate, ammonia excretion rate, and the ratio of oxygen to nitrogen (O:N) were examined after treatment with pedunsaponin. The correlations of these processes with pedunsaponin concentrations, treatment temperatures and snail sizes were also evaluated in this study.
The results showed that both the oxygen consumption rate and ammonia excretion rate decreased in the pedunsaponintreated groups compared with the untreated group (P＜0.05). When the temperature was 28 ℃ under 40 mg/L pedunsaponin, the oxygen consumption and ammonia excretion rate were 0.19 mg/(g·h) and 0.02 mg/(g·h) in the treated snails, while for the normal snails, they were 0.88 mg/(g·h) and 0.07 mg/(g·h), respectively. In addition, pedunsaponin significantly reduced the O:N ratio of the snails, shifting away from protein, fat and carbohydrate metabolism to main protein one.
In conclusion, this study demonstrates that pedunsaponin is able to reduce the level of respiratory metabolism of P. canaliculata, which may inhibit fat and carbohydrate metabolism, thus forcing the snail to increase protein metabolism for an immune response. As a result, the snails died due to the lack of an energy supply. The study provides a theoretical basis for further research into toxic mechanism of pedunsaponin on P. Canaliculata.

Phytoremediation is a low-cost and eco-friendly technology by using plants to uptake and accumulate heavy metals from contaminated soils. Intercropping is a simple and feasible method of agricultural management which was applied here to increase plant production and facilitate the efficiency of phytoremediation.
A pot experiment based on an intercropping system of moso bamboo seedlings and Sedum plumbizincicola was conducted to investigate its influence on the growth of plants, transfer ability of heavy metals in the system of soil and plant, and the effect on accumulation of Cu, Cd and Zn in different parts of plant.
The results showed that the root biomass of S. plumbizincicola which was intercropped with moso bamboo in 2 mm soil decreased significantly, and the biomass of both stems and leaves of moso bamboo which was intercropped with S. plumbizincicola in 5 mm soil was also declined clearly compared with that of moso bamboo in 2 mm soil. Moreover, the biomass of moso bamboo seedlings and S. plumbizincicola was reduced as compared with those of CK (monocropping), which suggested that intercropping led to competition between moso bamboo seedlings and S. plumbizincicola. Intercropping resulted in the decline of heavy metal uptake for the roots of moso bamboo seedlings and the competition between Cd and Zn in the roots of S. plumbizincicola, while
it improved both upward transport by roots, but there were differences in the process for different metals accumulated in different parts (stem or leaf) of moso bamboo seedlings. In addition, contrast to monocropping, Cu, Cd and Zn accumulation in any part of moso bamboo seedlings decreased distinctly, but Cd and Zn accumulation in the overground part of S. plumbizincicola increased by 57% and 82% in 5 mm soil.
In sum, the intercropping system of moso bamboo and S. plumbizincicola shows the potential of upward transfer of heavy metals, and it also has positive impacts on Cu, Cd and Zn accumulation.

The relationship between plants and environment is mainly reflected in the life history, morphology, phenology, physiology of plants. Different kinds of plants have different adaptation strategies. The southeastern of Qinghai-Tibet Plateau is among the highest forest line distribution areas and one of the most obvious climate change areas in the world. Understanding ecological and climatic indicators of the forest line in this area can offer a foundation for analyzing
the possible changes in the future climate scenario. By means of research on the basic characteristics and physiological indicators of Abies georgei var. smithii seedlings on Mt. Shergyla in southeastern Tibet, we hope to discover the response of Abies georgei var. smithii seedlings to the environment with different altitude gradients, and provide basic evidence for further explaining the response of Abies georgei var. smithii seedlings to altitude.
In this paper, we collected 350 seedlings of 7-8 years with great growth condition in the range of 3 800-4 400 m above sea level in April 2014. We measured their basic growth indexes, root activity, chlorophyll content, etc., and employed Excel 2013, R3.3.1, SPSS 21.0 and other software to analyze the data.
The results showed that: 1) The optimal growth of Abies georgei var. smithii seedlings was observed at the altitude of 3 900 m. The basic characteristics and chlorophyll content were significantly higher than the other altitudes. 2) At the altitude of 3 800 m to 4 100 m, the soluble protein content, soluble sugar content and root activity of Abies georgei var. smithii seedlings were basically stable, indicating that the environmental stress had no significant effect on the growth of Abies georgei var. smithii seedlings. 3) From the altitude of 4 200 m in the Mt. Shergyla, transition began with the forest line and tree line, and the external environment at the altitude of 4 400 m was harsh. The indexes such as seedling height, root activity and chlorophyll content declined sharply which were greatly affected by environmental stress.
In sum, the growth of Abies georgei var. smithii seedlings is largely affected by the altitude of Mt. Shergyla. The seedlings could not resist the effects of environmental stress in the range of the forest line, which resulted in short seedlings with weak energy and slow growth.

Diarrhea is a common disorder in pigs, and associated dehydration is a leading cause of mortality among piglets, causing substantial economic losses. Studies have found that the etiology of diarrhea is varied, including multiple viral, bacterial, and parasitic pathogens, but viruses such as porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV), rotavirus (RV) and other enteroviruses (EV), are the predominant factors in most cases. Repeated outbreaks have also been reported on farms in which piglets had been immunized by vaccines targeting these viruses. Recently, newly emerging viruses, such as mammalian orthoreovirus (MRV), which can cause diarrhea alone or in co-infections with other known pathogens, have been of great concern in China, South Korea, USA and Europe. Few researches on MRV, especially its epidemiological survey and analysis have been carried out in China.
In order to understand the epidemicity of MRV in China, molecular and seroepidemiological survey of MRV type 3 in diarrheic pigs from five provinces of east China was performed in this study. A nested reverse transcription-polymerase chain reaction (RT-PCR) method was designed to target the L1 gene. A total of 224 diarrheic fecal samples collected from 58 pig farms from Zhejiang, Jiangxi, Shandong, Henan and Heilongjiang provinces in 2013—2014 were examined by this method. Our data revealed that porcine orthoreovirus was present in 147 of 224 (65.6％) diarrheic porcine fecal samples obtained from 55 of 58 (94.8％) pig farms. At the same time, a neutralization test as well as an indirect enzyme-linked immunosorbent assay (ELISA) with a purified σ1 protein as a coat antigen was developed. The neutralization antibody levels and the anti-σ1 IgG antibody levels of 37 serum samples were determined. A good linear relationship between the anti-σ1 antibodies and virus neutralization (VN) antibodies was observed, demonstrating the correlation between them. Then the σ1- based ELISA was applied to the subsequent seroepidemiological survey of MRV3. A total of 262 serum samples collected from the sows, one- week-old piglets as well as three-week-old weaning piglets in 2015—2016 were tested. The result showed these samples had a high positive rate of anti-MRV IgG (84.4％).
Overall, the preliminary results suggest that MRVs are widespread in piglets from five provinces of east China and may collectively contribute to the enteric disease along with other porcine pathogens.

Macrobrachium rosenbergii is one of the major aquaculture shrimp, whose larva is the susceptible host of Macrobrachium rosenbergii Taihu virus (MrTV) which is one kind of novel RNA virus, and a member of Dicistroviridae owning a single positive strand with two open reading frames. Glutamate dehydrogenase (GDH) is a key enzyme in glutamate biosynthesis and plays an important role in organism’s metabolism. But there are still few reports about glutamate dehydrogenase in crustaceans, and almost no reports of M. rosenbergii glutamate dehydrogenase (MrGDH), as well as the interaction between MrGDH and MrTV.
In this study, the complete sequence of MrGDH gene was cloned by rapid amplification of cDNA ends (RACE) technology, and the expression of MrGDH gene in different tissues was studied by quantitative real-time polymerase chain reaction (qRT-PCR). Then the transcriptional characteristics of MrGDH gene in the gill and hepatopancreas of M. rosenbergii were analyzed under MrTV infection (positive group), and the negative group was injected with phosphate buffered solution.
The results showed that the complete cDNA sequence of MrGDH gene was 2 257 bp in length with a 1 662 bp open reading frame (ORF) encoding 553 amino acids, which shared 92% homology with Fenneropenaeus chinensis GDH and Litopenaeus vannamei GDH. The protein molecular mass of MrGDH was approximately 61.37 kDa. Its secondary and threedimensional structures were extremely similar to Drosophila melanogaster GDH and Bombyx mori GDH. The MrGDH gene was expressed in all detected tissues of M. rosenbergii. After 48 h and 72 h of injection, the expression of MrGDH gene in hepatopancreas and gill of positive group was significantly higher than the negative group.
In summary, the MrGDH gene is conservative, and its expression distribution indicates that it’s highly expressed in the organisms extremely related to movement and metabolism, with the highest in muscle and the lowest in haemolymph. This difference may be caused by the diversity in energy demand of each organization. Overall, it is indicated that MrTV infection could stimulate the expression of MrGDH gene.

Percent ground cover of vegetation is an important parameter which received attention of both agronomists and ecologists. Not only does it reflect dynamic growth of plants in a long time, but also it is associated with abstraction of photosynthesis available radiation (APAR) of plants. So far as the maize crop cover is concerned, current researches mainly focused on calculating percent ground cover of maize on bare ground. It is a fact that plastic film mulching has been widely adopted for maize planting due to its effect on reducing water loss, regulating soil temperature, improving the infiltration of rainwater into the soil, enhancing soil water retention, accelerating crop growth, and significantly increasing crop yield. In addition, the recent advances in image analysis software offered potential for analyzing the digital camera images of habitat to objectively quantify ground cover of vegetation in a repeatable and timely manner too. Here we evaluated use of Matlab software for analyzing the digital photographs of plastic-film maize to quantify the percent ground cover.
In this study, the images of plastic-film maize were firstly taken by smart phone under weak light condition, which were JPEG (joint photographic expert group) format here and were in 1 358×1 314 resolution. Then the method combined the K-mean clustering analysis of hue (H) and saturation (S) color components with performing a corresponding mathematical operation was proposed to discriminate the maize and background. The proposed method was comprised of three main steps. First, color images yielding red (R), green (G), and blue (B) subimages were mathematically transformed to hue (H), saturation (S), and intensity (I) ones. And then, the images were respectively segmented using the methods of excess green and excess green minus excess red. Second, the K-mean clustering analysis of H and S color components was carried out. Finally, the color difference operation between the K-mean clustering analysis of H and S color components was performed for segmentation of target object.
Results of images processing indicated that the images, which were segmented respectively by excess green, excess red, excess green minus excess red, and Otsu thresholding of excess green, excess red and excess green minus excess red, showed incomplete construct of maize and plastic film, but relatively satisfactory results were achieved by clustering analysis of H and S color components. Specifically, the K-mean clustering analysis of H color component clearly delineated leaf edge of maize, and the K-mean clustering analysis of S color component produced complete plastic film construct. The maize plant was successfully separated from plastic film, soil and other backgrounds by application of the color difference operation between the K-mean clustering analysis of H and S color components. Root mean square error (RMSE) and error rate were calculated to verify the reliability of the method proposed in this paper for segmentation of maize plant. The results showed that the RMSE and error rate of segmentation were 0.004 2 and 3.37%, respectively. The low RMSE and error rate further confirmed the rationality of the method used in this paper.
In conclusion, the method presented in this paper for image segmentation of plastic-film corn canopy is reliable under the weak light condition.