Calcium (Ca²⁺) plays an important role in determining plant growth and development because it maintains cell wall and
membrane integrity. Therefore, understanding the role of Ca2+ in carbon and lipid metabolism could provide insights
into the dynamic changes in cell membranes and cell walls during the rapid elongation of cotton fibers. In the present
study, we found that the lack of Ca²⁺ promoted fiber elongation and rapid ovule expansion, but it also caused tissue
browning in the ovule culture system. RNA-sequencing revealed that Ca²⁺ deficiency induced cells to be highly oxidized,
and the expression of genes related to carbon metabolism and lipid metabolism was activated significantly. All gene
members of nine key enzymes involved in glycolysis were up-regulated, and glucose was significantly reduced in Ca²⁺
deficiency-treated tissues. Ca²⁺ deficiency adjusted the flowing of glycolysis metabolic. However, low K⁺ recovered
the expression levels of glycolysis genes and glucose content caused by Ca2+ deficiency. Electrospray ionizationtandem
mass spectrometry technology was applied to uncover the dynamic profile of lipidome under Ca²⁺ and K⁺
interacted conditions. Ca²⁺ deficiency led to the decrease of fatty acid (FA), diacylglycerol (DAG), glycolipid and the
significant increase of triacylglycerol (TAG), phospholipid phosphatidylethanolamine (PE), phosphatidylglycerol (PG),
and PC (phosphatidylcholine). Low K⁺ restored the contents of FA, phospholipids, and glycolipids, effectively relieved
the symptoms caused by Ca²⁺ deficiency, and recovered the development of fiber cells. This study revealed dynamic
changes in transcript and metabolic levels and uncovered the signaling interaction of Ca²⁺ deficiency and low K⁺ in
glycolysis and lipid metabolism during fiber development.

Fertilization is an effective technique to improve soil fertility and increase crop yield. The long-term effects of different fertilizers on soil considerably vary. Over 38 consecutive years of different fertilization positioning experiments in a double cropping rice field of Qiyang Red Soil Experimental Station, seven different fertilization treatments including CK (no fertilization), NPK (nitrogen, phosphorus, and potassium fertilizer), M (cow manure), NPKM (nitrogen, phosphorus, and potassium with cow manure), NPM (nitrogen and phosphorus with cow manure), NKM (nitrogen and potassium with cow manure), and PKM (phosphorus and potassium with cow manure) were applied to study the effects on rice yield, soil fertility, and nutrient apparent balance in a paddy field. The results showed that the annual average yields of rice in NPKM, NPM, NKM, PKM, M, NPK and CK treatments ranged from 6 214 to 11 562 kg ha^–1. Yields under longterm organic and inorganic treatments (NPKM, NPM, NKM and PKM) were 22.58, 15.35, 10.53 and 4.41%, respectively, greater than under the NPK treatment. Soil organic carbon (SOC), total nitrogen (TN), available nitrogen (AN) and available potassium (AK) concentration with long-term organic and inorganic treatment (NPKM, NPM, NKM and PKM) were significantly higher than in inorganic fertilizer (NPK) treatments. Soil total phosphorus (TP) and available phosphorus (AP) contentration with organic fertilizer combined with inorganic N and P fertilizer treatment (NPKM, NPM and PKM) were significantly higher than with inorganic fertilizer alone (NPK treatments). The average annual rice yield (11 562 kg ha^–1), SOC (20.88 g kg^–1), TN (2.30 g kg^–1), TP (0.95 g kg^–1), TK (22.50 g kg^–1) and AP (38.94 mg kg^–1) concentrations were the highest in the NPKM treatment. The soil AN concentration (152.40 mg kg^–1) and AK contentration (151.00 mg kg^–1) were the highest in the NKM treatment. N and P application led to a surplus of nitrogen and phosphorus in the soil, but NPKM treatment effectively reduced the surplus compared with other treatments. Soils under all treatments were deficient in potassium. Correlation analysis showed that SOC, TN, AN, TP, and AP contentration was significantly correlated with rice yield; the correlation coefficients were 0.428, 0.496, 0.518, 0.501, and 0.438, respectively. This study showed that the combined application of N, P, and K with cow manure had important effects on rice yield and soil fertility, but balanced application of N, P, and K with cow manure was required.

Exploring the effects of sowing date and ecological points on the yield of semi-winter wheat is of great significance.  This study aims to reveal the effects of sowing date and ecological points on the climate resources associated with wheat yield in the Rice–Wheat Rotation System.  With six sowing dates, the experiments were carried out in Donghai and Jianhu counties, Jiangsu Province, China using two semi-winter wheat varieties as the objects of this study.  The basic seedlings of the first sowing date (S1) were planted at 300×10⁴ plants ha⁻¹, which was increased by 10% for each of the delayed sowing dates (S2–S6).  The results showed that the delay of sowing date decreased the number of days, the effective accumulated temperature and the cumulative solar radiation in the whole growth period.  The yields of S1 were higher than those of S2 to S6 by 0.22–0.31, 0.5–0.78, 0.86–0.98, 1.14–1.38, and 1.36–1.59 t ha^–1, respectively.  For a given sowing date, the growth days increased as the ecological point was moved north, while both mean daily temperature and effective accumulative temperature decreased, but the cumulative radiation increased.  As a result, the yields at Donghai County were 0.01–0.39 t ha^–1 lower than those of Jianhu County for the six sowing dates.  The effective accumulative temperature and cumulative radiation both had significant positive correlations with yield.  The average temperature was significantly negatively correlated with the yield.  The decrease in grain yield was mainly due to the declines in grains per spike and 1 000-grain weight caused by the increase in the daily temperature and the decrease in the effective accumulative temperature.

Cereal and legume intercropping has been widely adopted to increase crop productivity in sustainable farming systems worldwide.  Among different intercropping combinations, millet and peanut intercropping can be adapted to most water-limited areas.  However, there are few studies on the differences in yield characteristics and nitrogen use efficiency between millet/peanut intercropping and monocultures under different nitrogen (N) application rates.  The objective of this study was to determine the yield advantages and economic benefits, as well as the appropriate N application rate, of millet/peanut intercropping.  A two-year field experiment was conducted with three cropping patterns (monoculture millet, monoculture peanut and millet/peanut intercropping) and four N rates (0, 75, 150 and 225 kg ha⁻¹).  The results showed that the land equivalent ratio (LER) and net effect (NE) of the intercropping system reached their highest levels at the N input of 150 kg ha⁻¹ in 2018 and 2019 (1.04 for LER, 0.347 Mg ha⁻¹ for NE, averaged across two years).  Millet was the dominant crop in the intercropping system (aggressivity of millet and peanut (Amp)>0, competitive ratio of millet and peanut (CRmp)>1), and millet yields achieved their highest values at N inputs of 225 kg ha⁻¹ for monoculture and 150 kg ha⁻¹ for intercropping.  NUE reached its highest levels with N inputs of 150 kg ha⁻¹ for all planting patterns over the two years.  Intercropping combined with an N input of 150 kg ha⁻¹ achieved the highest net income of 2 791 USD ha⁻¹, with a benefit-cost ratio of 1.56, averaged over the two years.  From the perspective of economics and agricultural sustainable development, millet/peanut intercropping at 150 kg N ha⁻¹ seems to be a promising alternative to millet or peanut monoculture.

Rice, the main food crop in China, has been sporadically reported to suffer from weedy rice infestation.  However, the overall occurrence and distribution pattern of Chinese weedy rice remains unclear because a systematic survey has not been conducted.  In order to reveal the infestation of Chinese weedy rice, a field survey was conducted in 999 sampling sites all over the rice-growing regions in China from 2009 to 2016 using seven-scale visual scoring of the level of weed infestation.  Weedy rice was found 39% occurrence incidence in a total of 387 sites.  The sampling sites with 50% or higher overall weedy rice infestation index mainly radiated from Jiangsu, Heilongjiang, Ningxia and Guangdong to the whole East China, Northeast China, Northwest China and South China.  A total of 45 morphological characters from 287 populations (collected simultaneously with the field survey) out of those occurred sites were observed and analyzed using multivariate analysis in common gardens with the same cultivation conditions in 2017 and 2019.  Canonical correlation analysis showed that 45 morphological characters were significantly related to the latitude, mean temperature, minimum temperature, precipitation and mean diurnal range factors.  The 287 weedy rice populations were divided into three morphological groups with climate-dependent geographical differentiation: strong tiller type only in Jiangsu, large leaf type in South China and Central China and large grain type mainly in North China.  Weedy rice seriously infested rice fields and had a geography, climate and cultivated rice type-dependent morphological and biotype differentiation in China.  It is suggested to pay attention to the harmfulness of weedy rice and adopt comprehensive control strategies.

Primary porcine tracheal epithelial cells (PTECs) are an appropriate model for studying the molecular mechanism of various porcine respiratory diseases, including swine-origin mycoplasmas, which are isolated from respiratory tract of pigs and mainly found on the mucosal surface surrounding swine trachea.  However, the short proliferation ability of primary PTECs greatly limits their lifespan.  In this study, primary PTECs were carefully isolated and cultured, and immortal PTECs were constructed by transfecting primary PTECs with the recombinant constructed plasmid pEGFP-hTERT containing human telomerase reverse transcriptase (hTERT).  Immortal PTECs (hTERT-PTECs) maintained both the morphological and functional characteristics of primary PTECs, as indicated by the expression of cytokeratin 18, cell-cycle analysis, proliferation assay, Western blotting, telomerase activity assay, karyotype analysis and quantitative RT-PCR.  Compared to primary PTECs, hTERT-PTECs had an extended replicative lifespan, higher telomerase activity, and enhanced proliferative activity.  In addition, this cell line resulted in a lack of transformed and grown tumors in nude mice, suggesting that it could be safely applied in further studies.  Moreover, hTERT-PTECs were vulnerable to all swine-origin mycoplasmas through quantitative analysis as indicated by 50% color changing unit (CCU₅₀) calculation, and no significant differences of adhesion ability between primary and immortal PTECs were observed.  For the representative swine mycoplasma Mycoplasma hyopneumoniae (Mhp), except for DNA copies quantitative real-time PCR assay, indirect immunofluorescence assay and Western blotting analysis also depicted that hTERT-PTECs was able to adhere to different Mhp strains of different virulence.  In summary, like primary PTECs, hTERT-PTECs could be widely used as an adhesion cell model for swine-origin mycoplasmas and in infection studies of various porcine respiratory pathogens.  

Available irrigation resources are becoming increasingly scarce in the North China Plain (NCP), and nitrogen-use efficiency of crop production is also relatively low. Thus, it is imperative to improve the water-use efficiency (WUE) and nitrogen fertilizer productivity on the NCP. Here, we conducted a two-year field experiment to explore the effects of different irrigation amounts (S60, 60 mm; S90, 90 mm; S120, 120 mm; S150, 150 mm) and nitrogen application rates (150, 195 and 240 kg ha^–1; denoted as N1, N2 and N3, respectively) under micro-sprinkling with water and nitrogen combined on the grain yield (GY), yield components, leaf area index (LAI), flag leaf chlorophyll content, dry matter accumulation (DM), WUE, and nitrogen partial factor productivity (NPFP). The results indicated that the GY and NPFP increased significantly with increasing irrigation amount, but there was no significant difference between S120 and S150; WUE significantly increased first but then decreased with increasing irrigation and S120 achieved the highest WUE. The increase in nitrogen was beneficial to improving the GY and WUE in S60 and S90, while the excessive nitrogen application (N3) significantly reduced the GY and WUE in S120 and S150 compared with those in the N2 treatment. The NPFP significantly decreased with increasing nitrogen rate under the same irrigation treatments. The synchronous increase in spike number (SN) and 1 000-grain weight (TWG) was the main reason for the large increase in GY by micro-sprinkling with increasing irrigation, and the differences in SN and TGW between S120 and S150 were small. Under S60 and S90, the TGW increased with increasing nitrogen application, which enhanced the GY, while N2 achieved the highest TWG in S120 and S150. At the filling stage, the LAI increased with increasing irrigation, and greater amounts of irrigation significantly increased the chlorophyll content in the flag leaf, which was instrumental in increasing DM after anthesis and increasing the TGW. Micro-sprinkling with increased amounts of irrigation or excessive nitrogen application decreased the WUE mainly due to the increase in total water consumption (ET) and the small increase or decrease in GY. Moreover, the increase in irrigation increased the total nitrogen accumulation or contents (TNC) of plants at maturity and reduced the residual nitrate-nitrogen in the soil (SNC), which was conducive to the increase in NPFP, but there was no significant difference in TNC between S120 and S150. Under the same irrigation treatments, an increase in nitrogen application significantly increased the residual SNC and decreased the NPFP. Overall, micro-sprinkling with 120 mm of irrigation and a total nitrogen application of 195 kg ha–1 can lead to increases in GY, WUE and NPFP on the NCP.

Tillage represents an important practice that is used to dynamically regulate soil properties, and affects the grain production process and resource use efficiency of crops. The objectives of this 3-year field study carried out in the Huang-Huai-Hai (HHH) Plain of China were to compare the effects of a new deep vertical rotary tillage (DVRT) with the conventional shallow rotary tillage (CT) on soil properties, winter wheat (Triticum aestivum L.) grain yield and water and nitrogen use efficiency at different productivity levels, and to identify a comprehensive management that optimizes both grain yield and resource use efficiency in the HHH Plain. A split-plot design was adopted in field experiments in the winter wheat growing seasons of 2016–2017 (S1), 2017–2018 (S2) and 2018–2019 (S3), with DVRT (conducted once in June 2016) and CT performed in the main plots. Subplots were treated with one of four targeted productivity level treatments (SH, the super high productivity level; HH, the high productivity and high efficiency productivity level; FP, the farmer productivity level; ISP, the inherent soil productivity level). The results showed that the soil bulk density was reduced and the soil water content at the anthesis stage was increased in all three years, which were due to the significant effects of DVRT. Compared with CT, grain yields, partial factor productivity of nitrogen (PFP_N), and water use efficiency (WUE) under DVRT were increased by 22.0, 14.5 and 19.0%. Path analysis and direct correlation decomposition uncovered that grain yield variation of winter wheat was mostly contributed by the spike numbers per area under different tillage modes. General line model analysis revealed that tillage mode played a significant role on grain yield, PFP_N and WUE not only as a single factor, but also along with other factors (year and productivity level) in interaction manners. In addition, PFP_N and WUE were the highest in HH under DVRT in all three growth seasons. These results provided a theoretical basis and technical support for coordinating the high yield with high resource use efficiency of winter wheat in the resource-restricted region in the HHH Plain of China.

Northeast China (NEC) is one of the major maize production areas in China. Agro-climatic resources have obviously changed, which will seriously affect crop growth and development in this region. It is important to investigate the contribution of climate change adaptation measures to the yield and resource use efficiency to improve our understanding of how we can effectively ensure high yield and high efficiency in the future. In this study, we divided the study area into five accumulated temperature zones (ATZs) based on growing degree days (GDD). Based on the meteorological data, maize data (from agro-meteorological stations) and the validated APSIM-Maize Model, we first investigated the spatial distributions and temporal trends of maize potential yield of actual planted cultivars, and revealed the radiation use efficiency (RUE) and heat resource use efficiency (HUE) from 1981 to 2017. Then according to the potential growing seasons and actual growing seasons, we identified the utilization percentages of radiation (P_R) resource and heat resource (P_H) for each ATZ under potential production from 1981 to 2017. Finally, we quantified the contributions of cultivar changings to yield, P_R and P_H of maize. The results showed that during the past 37 years, the estimated mean potential yield of actual planted cultivars was 13 649 kg ha^–1, ranged from 11 205 to 15 257 kg ha^–1, and increased by 140 kg ha^–1 per decade. For potential production, the mean values of RUE and HUE for the actual planted maize cultivars were 1.22 g MJ^–1 and 8.58 kg (°C d)^–1 ha^–1. RUE showed an increasing tendency, while HUE showed a decreasing tendency. The lengths of the potential growing season and actual growing season were 158 and 123 d, and increased by 2 and 1 d per decade. P_R and P_H under potential production were 82 and 86%, respectively and showed a decreasing tendency during the past 37 years. This indicates that actual planted cultivars failed to make full use of climate resources. However, results from the adaptation assessments indicate that, adoption of cultivars with growing season increased by 2–11 d among ATZs caused increase in yield, P_R and P_H of 0.6–1.7%, 1.1–7.6% and 1.5–8.9%, respectively. Therefore, introduction of cultivars with longer growing season can effectively increase the radiation and heat utilization percentages and potential yield.

MicroRNAs (miRNAs) are endogenous non-protein-coding small RNAs that play crucial and versatile regulatory roles in plants.  Using a computational identification method, we identified 55 conserved miRNAs in tea (Camellia sinensis) by aligning miRNA sequences of different plant species with the transcriptome library of tea strain 1005.  We then used quantitative real-time PCR (qRT-PCR) to analyze the expression of 31 identified miRNAs in tea leaves of different ages, thereby verifying the existence of these miRNAs and confirming the reliability of the computational identification method.  We predicted which miRNAs were involved in catechin synthesis using psRNAtarget Software based on conserved miRNAs and catechin synthesis pathway-related genes.  Then, we used RNA ligase-mediated rapid amplification of cDNA ends (RLM-RACE) to obtain seven miRNAs cleaving eight catechin synthesis pathway-related genes including chalcone synthase (CHS), chalcone isomerase (CHI), dihydroflavonol 4-reductase (DFR), anthocyanidin reductase (ANR), leucoanthocyanidin reductase (LAR), and flavanone 3-hydroxylase (F3H).  An expression analysis of miRNAs and target genes revealed that miR529d and miR156g-3p were negatively correlated with their targets CHI and F3H, respectively.  The expression of other miRNAs was not significantly related to their target genes in the catechin synthesis pathway.  The RLM-RACE results suggest that catechin synthesis is regulated by miRNAs that can cleave genes involved in catechin synthesis. 

A total of 50 endophytic bacterial isolates were obtained from Kobreasia capillifolia at alpine grasslands in the Eastern Qilian Mountains on the Tibetan Plateau in China.  Based on the sequencing and phylogenetic analysis of 16S rDNA genes, all isolates phylogenetically related closely to Bacillus, Acinetobacter, Stenotrophomonas, Brevundimonas, Arthrobacter, Curtobacterium, Paenibacillus, Plantibacter, Promicromonospora, Serratia, and Microbacterium, among which Bacillus was the predominant genus (47.8% of the total number of endophytic isolates).  These isolates possessed different biological functions.  In 50 endophytic bacteria, 42 isolates produced indole acetic acid (IAA) on King medium.  There were seven isolates showing potency of mineral phosphate solubilization in Pikovaskaia’s (PKO) liquid medium.  Seven isolates exhibited antagonistic effect against Fusarium avenaceum, Colletotrichum coccodes and Phoma foveata.  This was the first report on diversity and plant growth promotion of endophytic bacteria from K. capillifolia on alpine grassland in the Eastern Qilian Mountains, Chain.  It is essential for revealing the relationship among plant, microorganism, and the special environment because the potential function of endophytic bacteria made a contribution to the interactions of plants and endophytic bacteria.

    Pepper (Capsicum spp.) is an important vegetable crop in the world. Now the pepper in China contributes one-third of the world’s peppers production. Genetic diversity of the pepper germplasm of China is expected interesting to know. To explore the structure of genetic diversity in Chinese pepper germplasm resources and possible relationship with cultivar types or geographic origin, we sampled and compared 372 GenBank pepper accessions (local cultivars and landraces) from 31 provinces, autonomous regions and municipalities of China and 31 additional accessions from other countries. These accessions were genotyped using 28 simple sequence repeat (SSR) markers spanning the entire pepper genome. We then investigated the genetic structure of the sampled collection using model-based analysis in STRUCTURE v2.3.4 and examined genetic relationships by the unweighted pair-group method of mathematical averages (UPGMA) in MEGA. In addition to geographic origin, we evaluated eight plant and fruit traits. In total, 363 alleles were amplified using the 28 SSR primers. Gene diversity, polymorphism information content and heterozygosity of the 28 SSR loci were estimated as 0.09–0.92, 0.08–0.92 and 0.01–0.34, respectively. The UPGMA cluster analysis clearly distinguished Capsicum annuum L. from other cultivated pepper species. Population structure analysis of the 368 C. annuum accessions uncovered three genetic groups which also corresponded to distinct cultivar types with respect to the plant and fruit descriptors. The genetic structure was also related to the geographic origin of the landraces. Overall results indicate that genetic diversity of Chinese pepper landraces were structured by migration of genotypes followed by human selection for cultivar types in agreement with consumption modes and adaptation to the highly diversified agro-climatic conditions.

Fruit set and development are affected by many phytohormones, including gibberellin.  Little is known regarding molecular mechanism underlying gibberellin mediated fruit set and development especially in Capsicum.  Three gibberellin receptors, CaGID1b.1, CaGID1b.2 and CaGID1c, and a DELLA protein, CaGAI, have been identified in Capsicum annuum L.  During the fruit development, the expression level of CaGID1c was low, and the expression fold change is mild.  However, CaGID1b.1 and CaGID1b.2 were relatively higher and more acute, which indicates that CaGID1b.1 and CaGID1b.2 may play an important role in fruit pericarp, placenta and seed.  Ectopic expressions of CaGID1b.1, CaGID1b.2 and CaGID1c in Arabidopsis double mutant gid1a gid1c increased plant height, among which CaGID1b.2 had the most significant effect; CaGAI reduced plant height in double mutant rga-24/gai-t6, having a similar function to AtGID1 and AtGAI in stem elongation.  Yeast two-hybrid (Y2H) and bimolecular fluorescence complementation (BiFC) assays indicated that CaGID1b.1 and CaGID1b.2 interact with CaGAI in a GA-dependent manner, while CaGID1c interacts with CaGAI in a GA-independent manner.  Our study reveals the key elements during gibberellin signaling in Capsicum and supports the critical importance of gibberellin for Capsicum fruit set and development.

In this study, four strains of Toxoplasma gondii with the same genetic type (Type I) originated from chicken, human, cat and swine were used to compare the immune responses in resistant chicken host to investigate the relationships between the parasite origins and the pathogenicity in certain host. A total of 300, 10-day-old chickens were allocated randomly into five groups which named JS (from chicken), CAT (from cat), CN (from swine), RH (from human) and a negative control group (–Ve) with 60 birds in each group. Tachyzoites of four different T. gondii strains (JS, CAT, CN and RH) were inoculated intraperitoneally with the dose of 1×107 in the four designed groups, respectively. The negative control (–Ve) group was mockly inoculated with phosphate-buffered saline (PBS) alone. Blood and spleen samples were obtained on the day of inoculation (day 0) and at days 4, 11, 25, 39 and 53 post-infection to screen the immunopathological changes. The results demonstrated some different immune characters of T. gondii infected chickens with that of mice or swine previous reported. These differences included up-regulation of major histocompatibility complex class II (MHC II) molecules in the early stage of infection, early peak expressions of interleukin (IL)-12 (IL-12) and -10 (IL-10) and long keep of IL-17. These might partially contribute to the resistance of chicken to T. gondii infection. Comparisons to chickens infected with strains from human, cat and swine, chickens infected with strain from chicken showed significant high levels of CD4+ and CD8+ T cells, interferon gamma (IFN-γ), IL-12 and IL-10. It suggested that the strain from chicken had different ability to stimulate cellular immunity in chicken.

Low light stress is one of the most important factors affecting photosynthesis and growth in winter production of cucumber (Cucumis sativus L.) in solar greenhouses in northern China. Here, two genotypes of cucumber (Deltastar and Jinyan 2) are used to determine the effect of low light stress on Rubisco expression and photosynthesis of leaves from emergence to senescence. During leaf development, the net photosynthetic rate (PN), stomatal conductance (gs), Rubisco initial activity and activation state, transcript levels of rbcL and rbcS, and the abundance of rbcL and rbcS DNA in these two genotypes increase rapidly to reach maximum in 10-20 d, and then decrease gradually. Meanwhile, the actual photosystem II efficiency (ФPSII) of cucumber leaves slowly increased in the early leaf developing stages, but it declined quickly in leaf senescent stages, accompanied by an increased non-photochemical quenching (NPQ). Moreover, PN, gs, initial Rubisco activity, and abundance of protein, mRNA and DNA of Rubisco subunits of leaves grown under 100 μmol m-2 s-1 are lower, and require more time to reach their maxima than those grown under 600 μmol m-2 s-1 during leaf development. All these results suggest that lower photosynthetic capacity of cucumber leaves from emergence to senescence under low light stress is probably due to down-regulated Rubisco gene expression in transcript and protein levels, and decreased initial and total activity as well as activation state of Rubisco. Deltastar performs better than Jinyan 2 under low light stress.

It is of great importance to solve the threats induced by cadmium pollution on crops. This paper examined the effect of biochar on cadmium accumulation in japonica rice and revealed the mechanism underlying the response of protective enzyme system to cadmium stress. Biochar derived from rice straw was applied at two application rates under three cadmium concentrations. Shennong 265, super japonica rice variety, was selected as the test crop. The results indicated that cadmium content in above-ground biomass of rice increased with increasing soil cadmium concentrations, but the biochar application could suppress the accumulation of cadmium to some extent. Under high concentrations of cadmium, content of free proline and MDA (malondialdehyde) were high, so did the SOD (superoxide dismutase), POD (peroxidase) and CAT (catalase) activity in the flag leaf of rice. However, the protective enzyme activities remained at low level when biochar was added.

Based on the sequence of a novel expressed sequence tag (EST), the full-length cDNA of 1 017 nucleotides was cloned from Brassica napus cv. Xiangyou 15 through rapid amplification of cDNA ends (RACE). The gene was designated as Bnhol34 (HQ585980), encoding a protein of 338 amino acids. BLAST analysis showed no high degree of sequence identity to any known gene. The calculated molecular weight of the Bnhol34 protein was 36.23 kDa, and the theoretical isoelectric point was 8.74. The Bnhol34 was also cloned from a high oleic acid mutant 854-1 through homologous cloning. There was no difference between the two Bnhol34 genes. Bnhol34 was localized in a tissue-specific manner in B. napus, and its expression level was about eight-fold greater in Xiangyou 15 seeds than in 854-1. The promoter region sequences of Bnhol34 were then isolated from Xiangyou 15 and 854-1, and a 93-bp deletion was found to occur in the Bnhol34 promoter region of 854-1. Three abscisic acid-responsive cis-elements (ABRE) were identified in the promoter region of Xiangyou 15. Real-time PCR analyses revealed that exogenous abscisic acid increased Bnhol34 expression by about four-fold in Xiangyou 15 seeds, yet did not change Bnhol34 expression in 854-1. It appeared that Bnhol34 might be abscisic acid insensitive in 854-1.

Low temperature and high salinity are the major abiotic stresses that restrict cucumber growth and production, breeding materials with multiple abiotic resistance are in greatly need. Here we investigated the effect of introducing the LOS5 gene, a key regulator of ABA biosynthesis in Arabidopsis thaliana, under the stress-responsive RD29A promoter into cucumber (Cucumis sativus L. cv. S516). We found that T1 RD29A-LOS5 transgenic lines have enhanced tolerance to cold and salt stresses. Specifically, transgenic lines exhibited dwarf phenotypes with reduced leaf number, shorter internode, decreased length of the biggest leaf, fewer female flowers, shorter fruit neck and lower vitamin C (Vc). The increased cold tolerance can be reflected from the significantly decreased cold index, the reduced electrolyte leakage index and the MDA content upon cold treatment as compared to those in the control. This may result from the accumulation of internal ABA, soluble sugars and proline, and the enhanced activities of protective enzymes superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) in the transgenic lines. Under salt treatment, the transgenic lines exhibited increased germination index, vigor index, more lateral roots and increased root fresh weight. Moreover, RD29A-LOS5 transgenic plants displayed quicker responses in salt stress than that in low-temperature stress.

Low light stress is one of the main limiting factors which influence the production of sweet pepper under protected cultivation in China. In this experiment, two genotypes of sweet pepper, ShY (low light-tolerant genotype) and 20078 (low light-sensitive genotype), were used to study the effects of low light (photosynthetic photon flux density, PPFD was 75- 100 μmol m-2 s-1, control 450-500 μmol m-2 s-1) on photosynthesis during leaf development. The result indicated that under low light chlorophyll content, net photosynthetic rate (PN), photosynthetic apparent quantum efficiency ( i) and carboxylation efficiency (CE) of sweet pepper leaves increased gradually and decreased after reaching the maximum levels. The time to reach the peak values for all the above parameters was delayed, whereas the light compensation point (LCP) decreased gradually along with leaf expansion. The decrease in maximum quantum yield of PS II (Fv/Fm) was not observed at any stages of the leaf development under low light condition, but the actual PS II efficiency under irradiance ( PS II) was lower accompanied by an increased non-photochemical quenching (NPQ) in young and/or old leaves compared with mature leaves. The antenna thermal dissipation (D) was a main way of heat dissipation when young leaves received excessive light energy, while the decline in photosynthetic function in senescence leaf was mostly owing to the decrease in carbon assimilation capacity, followed by a significantly increased allocation of excessive energy (Ex). Compared with 20078, ShY could maintain higher PN, PS II and lower QA reduction state for a longer time during leaf development. Thus, in ShY photosynthetic efficiency and the activity of electron transport of PS II were not significantly affected due to low light stress.

CatSper is a unique Ca2+ channel-like protein family exclusively expressed in the testis and sperm, and plays important roles in sperm motility, capacitation, acrosome reaction and sperm-egg interactions. Here we studied the mechanism of regulation of CatSper2-dependent Ca2+ influx, extracellular and intracellular Ca2+ on sperm hyperactivated motility. The siRNA duplexes were transfected into the sperm cells by electroporation (EP) to silence the expression of CatSper2. The results for targeted disruption of CatSper2 showed the highest silence efficiency 77.7% (P<0.05), the hyperactivated sperm rate calculated by computer-assisted semen analysis (CASA) analysis of interferenced sperm was significantly lower 11.1% than the control 99.2%. Flow cytometry (FCM) detection of the intracellular Ca2+ concentration of interferenced sperm was 50 nmol L-1 higher than the normal. It was remarkably lower than hyperactivated sperm with 200-1 000 nmol L-1 (P<0.05). It was not sufficient to evoke hyperactivation. To trigger hyperactivation, the sperm were incubated in 50 μmol L-1 thimerosal or 5 mmol L-1 procaine, it sharply increased the intracellular Ca2+ via two different channels. CASA and FCM detection indicated that intracellular Ca2+ is required for initiating hyperactivation while extracellular Ca2+ is essential to maintain the process. We concluded that to mediate sperm hyperactivation, it is essential to inhibit Ca2+ peak present with targeted disruption of CatSper2 expression. This provides important prospective for further exploration of signal channel of sperm hyperactivated motility, potential factors for male infertility and provide further reference to the exploration of male contraceptive drug targets of male reproduction.