Determination of critical nitrogen dilution curve based on leaf area index for winter wheat in the Guanzhong Plain,Northwest China

Excessive use of nitrogen(N) fertilizers in agricultural systems increases the cost of production and risk of environmental pollution. Therefore, determination of optimum N requirements for plant growth is necessary. Previous studies mostly established critical N dilution curves based on aboveground dry matter(DM) or leaf dry matter(LDM) and stem dry matter(SDM), to diagnose the N nutrition status of the whole plant. As these methods are time consuming, we investigated the more rapidly determined leaf area index(LAI) method to establish the critical nitrogen(N_c) dilution curve, and the curve was used to diagnose plant N status for winter wheat in Guanzhong Plain in Northwest China. Field experiments were conducted using four N fertilization levels(0, 105, 210 and 315 kg ha-1) applied to six wheat cultivars in the 2013–2014 and 2014–2015 growing seasons. LAI, DM, plant N concentration(PNC) and grain yield were determined. Data points from four cultivars were used for establishing the N_c curve and data points from the remaining two cultivars were used for validating the curve. The N_c dilution curve was validated for N-limiting and non-N-limiting growth conditions and there was good agreement between estimated and observed values. The N nutrition index(NNI) ranged from 0.41 to 1.25 and the accumulated plant N deficit(N_(and)) ranged from 60.38 to –17.92 kg ha~(-1) during the growing season. The relative grain yield was significantly affected by NNI and was adequately described with a parabolic function. The N_c curve based on LAI can be adopted as an alternative and more rapid approach to diagnose plant N status to support N fertilization decisions during the vegetative growth of winter wheat in Guanzhong Plain in Northwest China.     

Identification of an H1N1 subtype of swine influenza virus and serological analysis

To investigate the epizootic of swine influenza virus(SIV), 60 nasal swabs were collected from a clinical cases of pig farm in Tai'an City, Shandong Province of China in April 2017. SIV was isolated by inoculating into 10-day-old Special Pathogen Free embryonated eggs and the whole genome was sequenced. An H1N1 subtype SIV was isolated and designated as A/swine/Shandong/TA04/2017(H1N1). Phylogenetic analysis showed that apart from the polymerase A(PA) fragment belonging to the 2009 pandemic H1N1 branch, seven genome segments belonged to avian-like H1N1 influenza virus lineage. The cleavage site sequence of the hemagglutinin(HA) protein was PSIQSR↓G, which is a typical molecular biological characteristic. Five potential N-glycosylation sites(N14, N26, N277, N484 and N543) were found in the HA gene. To further investigate the epidemiology of SIV in this farm, the 995 serum samples were assessed with EAH1N1 2009 pandemic H1N1 and H3 N2 antigens. The results showed that the total positive rate was 65.43%. The positive rates of single virus infection detected by EAH1N1, 2009 pdmH1N1 and H3 N2 for serum HI(Hemagglutination inhibition) were 48.35, 30.85 and 7.47%, respectively. The results showed that SIV in Shandong Province has been reassorted in some segments and the SIV-positive rate was high on the SIV outbreak farm. These data provide evidence of an epizootic of SIV.     

Identification,characterization and full-length sequence analysis of a novel endornavirus in common sunflower (Helianthus annuus L.)

To identify the possible quarantine viruses in seven common sunflower varieties imported from the United States of America and the Netherlands, we tested total RNAs extracted from the leaf tissues using next-generation sequencing of small RNAs. After analysis of small RNA sequencing data, no any quarantine virus was found, but a double-stranded RNA(dsRNA) molecule showing typical genomic features of endornavirus was detected in two varieties, X3939 and SH1108. Full-length sequence and phylogenetic analysis showed that it is a novel endornavirus, temporarily named as Helianthus annuus alphaendornavirus(HaEV). Its full genome corresponds to a 14 662-bp dsRNA segment, including a 21-nt 5′ untranslated region(UTR), 3' UTR ending with the unique sequence CCCCCCCC and lacking a poly(A) tail. An open reading frame(ORF) that encodes a deduced 4 867 amino acids(aa) polyprotein with three domains: RdRP, Hel and UGT(UDP-glycosyltransferase). HaEV mainly distributed in the cytoplasm but less in the nucleus of leaf cells by fluorescence in situ hybridization(FISH) experiment. This virus has a high seed infection rate in the five varieties, X3907, X3939, A231, SH1108 and SR1320. To our knowledge, this is the first report about the virus of the family Endornaviridae in the common sunflower.     

Comparison of phenolic profiles and antioxidant activities in skins and pulps of eleven grape cultivars(Vitis vinifera L.)

Eleven grape cultivars were analysed to explore the variety differences of fresh grape phenolic profiles. The results showed that free phenolics were predominant in grape skins and pulps, and showed the higher antioxidant activities than bound. In 11 cultivars, Muscat Kyoho extracts had the highest total phenolic content in skins(10.525 mg GAE g~(–1) FW) and pulps(1.134 mg GAE g~(–1) FW), and exhibited the highest DPPH radical scavening capacity(EC_(50)=11.7 μg mL~(–1)) and oxygen radical absorbance capacity(ORAC) value(190.57 μmol TE g~(–1) FW) of free phenolic in skin. In addition, the most abundant phenolics in grape skins were found to be flavonoids such as kaempferol in Kyoho skin(541.2 μg g~(–1) FW), rutin, catechin and epicatechin in Muscat Kyoho skin(262.3, 86.3 and 70.0 μg g~(–1) FW, respectively). Furthermore, the principal component analysis showed a strong difference of phenolic profiles with the cultivars, existing forms and distributions. Pearson correlation coefficient analysis showed a significant linear correlation between total phenolic content and antioxidant activity(P0.05). Therefore, both skins and pulps were rich sources of bioactive phenolic compounds, and Muscat Kyoho was the ideal source among all samples.     

Effects of the severity and timing of basal leaf removal on the amino acids profiles of Sauvignon Blanc grapes and wines

The effects of the severity and timing of leaf removal(LR) on the amino acids of Sauvignon Blanc grapes and wines were studied during the 2017 growing season. High-performance liquid chromatography(HPLC) was used to analyze the amino acids profiles of grape berries and wines. The basal leaves were removed at three time points(40, 56 and 72 days after flowering, named LR40, LR56 and LR72, respectively) at two severity levels(one at which the first, third, and fifth basal leaves of each shoot were removed(50% level); and another at which the first six basal leaves were removed(100% level)). The results showed that leaf removal had little impact on total soluble solids(°Brix), titratable acidity, pH or berry weight. The LR72-50% treated grapes had higher berry weight, titratable acidity and °Brix than those of the other treatments. The highest concentrations of total amino acids and of total amino acids except proline were detected in LR72-50% treated grapes(2 952.58 and 2 764.36 mg L~(-1), respectively); the lowest were detected in LR72-100% treated grapes(2 172.82 and 2 038.71 mg L~(-1), respectively). LR72-50% treatment significantly promoted the synthesis of aspartic acid, serine, arginine, alanine, aminobutyric acid and proline at both severity levels for grapes, the concentrations of all of these amino acids were increased relative to the control concentrations. The LR72-50%, LR40-100% and LR72-100% treated wines had higher total amino acids concentrations and higher concentrations of some individual amino acids, such as arginine, alanine and serine, than did the control wines. Of all the amino acids studied, glycine, tyrosine, cysteine, methionine and lysine were not significantly influenced by the timing or severity basal defoliation in grapes and wines. The present study reveals the effects of the timing and severity of leaf removal on the amino acids profiles of grapes and wines.     

Reduction in cadmium accumulation in japonica rice grains by CRISPR/Cas9-mediated editing of OsNRAMP5

Cadmium (Cd) intake is harmful to human health and Cd contamination in rice grains represents a severe threat to those consuming rice as a staple food. Knockout of Cd transporters is a promising strategy to reduce Cd accumulation in rice grains. OsNRAMP5 is the major transporter for Cd and manganese (Mn) uptake in rice. Nevertheless, it is uncertain whether knockout of OsNRAMP5 is applicable to produce low Cd rice without affecting plant growth and grain yield. In this study, we adopted CRISPR/Cas9-based gene editing technology to knock out OsNRAMP5 in two japonica varieties. We generated three independent transgene-free osnramp5 mutants and investigated the effect of osnramp5 mutations on Cd accumulation and plant growth. Hydroponic experiments showed that plant growth and chlorophyll content were significantly reduced in osnramp5 mutants at low Mn conditions, and this defective growth in the mutants could be fully rescued by supply of high levels of Mn. Cd and Mn accumulation in both roots and shoots was markedly reduced in the mutants compared to that in wild-type plants. In paddy field experiments, although Cd in flag leaves and grains was greatly reduced in osnramp5 mutants, some agronomic traits including plant height, seed setting rate, and grain number per panicle were affected in the mutants, which ultimately caused a mild reduction in grain yield. The reduced plant growth in the mutants can be attributed to a marked decrease in Mn accumulation. Our results reveal that the manipulation of OsNRAMP5 should be treated with caution: When assessing the applicability of osnramp5 mutants, soil pH and soil water content in paddy fields need to be taken into consideration, since they might affect the levels of available Mn in the soil and consequently determine the effect of the mutation on grain yield.     

青海黄河沿岸土壤盐分特征及影响因素研究

应用常规统计学方法和典范对应分析方法,研究青海黄河沿岸不同程度盐渍化土壤各属性特征及其数量关系,探讨了土壤属性与环境因子之间的关系。结果表明:非农地土壤属中度、重度盐渍化土壤和盐土;随盐渍化程度增强,土壤阳离子从镁钙盐型转变为钠盐型;土壤阴离子从硫酸盐型转变为氯化物型;除Ca~(2+)与Cl~-、K~+相关性不显著外,其他各离子浓度之间显著正相关;有机质与pH值显著负相关;电导率与各离子浓度极显著正相关,与总盐浓度的相关性高达0.91;土壤各粒级含量及分形维数与各离子浓度、电导率、总盐浓度之间相关性不显著。土地利用类型能够很好地指示土壤盐渍化水平,盐渍化程度同时受微地形影响。土壤盐碱化与沙化是两个相关的土壤退化过程。     

Serologic and molecular survey for major viral pathogens in grazing hybrid wild boars in Northeast China

Hybrid wild boar husbandry is an important component of livestock production in Northeast China. However, the current disease situation of these animals is largely unknown due to a lack of disease surveillance. The present study was conducted to determine the prevalence of several important viral diseases in the hybrid wild boar population of Northeast China. Between September 2015 to December 2016, 169 blood and 61 tissue samples were collected from apparently healthy hybrid wild boars from farms in Jilin, Inner Mongolia and Heilongjiang provinces. ELISA detected serum antibodies against classical swine fever virus(CSFV), porcine reproductive and respiratory syndrome virus(PRRSV), pseudorabies virus(PRV), porcine circovirus type 2(PCV2) and Japanese encephalitis virus(JEV), but not against African swine fever virus(ASFV), with PCV2 having the highest seropositive rate(87.2–100% in different farms). RT-PCR or PCR performed on the processed samples detected only PCV2, with 33.1%(56/169) of blood samples and 32.8%(20/61) of spleen samples being positive, respectively, indicating widespread PCV2 infection in hybrid wild boars. Phylogenetic analysis of 15 PCV2 ORF2 sequences showed that they belong to genotypes PCV2a, PCV2b and PCV2d, with nucleotide and deduced amino acid homologies of 88.5–100% and 88.1–100%, respectively.     

东湖主要湖区浮游植物群落结构特征及其与环境因子的关系

[目的]了解东湖浮游植物的分布特征,为今后东湖的水资源开发与水环境保护提供生物学依据。[方法]2010年对武汉东湖4个主要子湖的浮游植物和环境因子进行每月调查。[结果]共检出浮游植物7门123种（绿藻58种,硅藻26种,蓝藻16种,裸藻14种,其他9种）,4个子湖中蓝藻、绿藻和硅藻的数量之和占总数量的98%以上,为东湖的优势藻门;东湖不同子湖的Shannon-W iener多样性指数均在1～3,属中污水质,湖泊水质为牛巢湖〉汤菱湖〉郭郑湖〉后湖;CCA排序分析表明东湖水体中浮游植物的空间分布主要受硝态氮和氨氮的影响,说明东湖是一个氮营养盐限制型湖泊;对比郭郑湖2010年与20世纪80年代的生物和理化指标,发现水体中硝态氮、氨氮含量和蓝绿藻的现存量都有显著提高。[结论]近年来,东湖从磷限制转变为氮限制型湖泊,富营养化正在日益加剧。     

Roles of pattern of secretion of luteinizing hormone and the ovary in attainment of puberty in ewes lambs

Four experiments were conducted to examine (1) how patterns of luteinizing hormone (LH) change as lambs approach first estrus, (2) whether mimicry of these changes by exogenous LH will initiate events of the pubertal process, (3) aspects of possible roles of the ovary and estradiol in the initiation of an ovulatory surge of LH in prepubertal lambs and (4) the time requirements for the presence of the ovary in induction of that surge of LH.In 19 lambs (Exp. 1), concentrations of LH in plasma were higher at puberty than 7 weeks earlier. Increased concentrations of LH in samples taken every 20 min for 6 hr during the luteal and follicular phases preceding first estrus were attributed to increased amplitude, but not increased frequency, of episodic pulses of LH. In Exp. 2, endocrine events which are included in the natural onset of ovarian cyclic activity (i.e. surge of LH and subsequent rise in progesterone) were initiated in $\text{5}\text{6}$, $\text{4}\text{6}$, $\text{1}\text{5}$, $\text{1}\text{6}$, and $\text{3}\text{6}$ lambs, receiving either injections of 7.5 μg/hr, 15 μg/hr, 30 μg/2hr, 45 μg/3hr or constant infusions of 15 μg/hr of purified ovine LH, respectively. However, these lambs did not exhibit estrus or cyclic ovarian activity. In Exp. 3, the mechanism by which hourly injections of LH prompted surges of LH was determined to be mediated through ovarian stimulation and not through a direct effect of exogenous LH on the hypothalamo-pituitary axis. All the intact (n=5) but no acutely (two weeks) ovariectomized (n=5) lambs had a preovulatory-like surge in response to exogenous LH. Further, (Exp. 4) it was shown that the ovarian signal must be maintained or evoked within 6 hr preceding the surge of LH, because lambs (n=20) ovariectomized at or 9, 18 or 27 hr after initiation of the exogenous LH with the one exception failed to show a surge of LH. It could not be demonstrated that the ovarian signal involved changes in peripheral concentrations of androstenedione, testosterone or estradiol-17β, although patterns of LH in ovariectomized lambs were responsive to both negative and positive feedback effects of estradiol (n=6, Exp. 3) and in general estradiol levels were increasing prior to the induced surge of LH.