Disaccharide combinations and the expression of enolase3 and plasma membrane Ca2+ ATPase isoform in sturgeon sperm cryopreservation

Acipenser sinensis and Acipenser dabryanus are critically endangered species, so germplasm conservation via cryopreservation of sperm is necessary. Disaccharides can act as membrane‐impermeable cryoprotectants, and enolase3 (ENO3) and plasma membrane Ca²⁺ ATPase isoform (PMCA2) are proteins associated with sperm quality. We considered seven characteristics of sperm quality in cultured brood stock from A. sinensis and A. dabryanus. We tested use of sucrose or trehalose alone and in combination at different concentrations for cryopreservation of A. dabryanus sperm. A low concentration of sucrose plus trehalose (S₁₅T₁₅) was optimal. Mixing of the extender with sucrose, lactose, or trehalose alone or with pairwise mixtures revealed that a mixture of lactose and trehalose (L₁₅T₁₅) gave the best results for both A. sinensis and A. dabryanus. Enolase3 and PMCA2 expression levels were measured in cryopreserved A. sinensis sperm via Western blotting. Relative ENO3 and PMCA2 expression levels were examined, and the relationship between disaccharide composition, sperm quality and protein expression was explored in A. sinensis. The results showed that relative ENO3 and PMCA2 expression levels were the highest at L₁₅T₁₅ in cryopreserved A. sinensis sperm. There were significant positive correlations between ENO3 expression and percentage membrane integrity, and between PMCA2 expression and sperm motility parameters (percentage of motile sperm, curvilinear velocity, straight‐line velocity and average path velocity; p < .05) in cryopreserved A. sinensis sperm. Our results indicate the optimal disaccharide combination and concentrations for cryopreservation of A. sinensis and A. dabryanus sperm and suggest that ENO3 and PMCA2 expression levels could serve as a valuable indicator of sperm quality in A. sinensis.     

Endocannabinoids as endometrial inflammatory markers in lactating Holstein cows

The objective of this study was to consider endocannabinoid system as inflammatory markers in bovine endometrium to better understand the role of this system in regulating many of the functions that are related to inflammatory condition. At day 26 post‐partum, fourteen cows were divided into two groups depending on the inflammatory condition: 1‐ subclinical endometritis (n = 7, with purulent or mucopurulent uterine discharge detectable in the vagina) and 2‐ healthy (n = 7, No (muco)) purulent discharge. Blood samples were collected at 26 and 30 days relative to calving to determine plasma tumour necrosis factor (TNF) and lipopolysaccharide‐binding protein (LBP) concentrations; moreover, uterine biopsy was carried out on day 26 post‐partum to measure mRNA abundance of TNF, interleukin‐1B (IL1B), interleukin‐6 (IL‐6), C‐X‐C motif chemokine ligand 8 (CXCL8), endocannabinoid receptor (CNR2), N‐acyl phosphatidylethanolamine phospholipase D (NAPEPLD), fatty acid amide hydrolase (FAAH), N‐acylethanolamine acid amidase (NAAA) and monoglyceride lipase (MGLL) by real‐time PCR. Results showed mean plasma concentrations of TNF and LBP were lower in healthy cows compared to subclinical endometritis cows (p < .05). Relative mRNA expression for NAAA and FAAH was decreased (p < .05), and relative mRNA expression for CNR2 and NAPEPLD increased in cows with subclinical endometritis compared to healthy cows. In conclusion, relative mRNA expression of TNF, IL1B and CXCL8 and plasma concentration of LBP increased during inflammatory condition along with decreased endocannabinoids hydrolyzing enzyme (NAAA and FAAH), increased enzymes that synthesize endocannabinoids (NAPEPLD) and relative gene expression of the endocannabinoid receptor; together, these contribute to increased endocannabinoids levels during inflammation. Overall, we provide evidence that endocannabinoid system is altered in endometrium tissue during inflammation through increased mRNA expression of CNR2 and synthesis enzyme and decreased mRNA expression of hydrolyzing enzymes interfere with pro‐cytokine production and signalling, which may interfere with the onset and progression of inflammation.     

热带海岸次生林群落主要乔木的种间关联分析

对热带海岸次生林群落主要乔木种群的重要值、方差比率、χ~2统计量和Ochiai(OI)指数等种间联结性指标进行测定,旨在揭示该群落优势种群之间相互作用的内在关联及种对间关联性的内在规律。结果表明:香蒲桃是热带海岸次生林群落的主要建群种;热带海岸次生林群落19个主要种群总体关联性表现为不显著正关联,反映了该群落尚未达到稳定的演替阶段;χ~2检验结果显示该群落优势种群正关联种对与负关联种对数量相当,无关联种对数较少,分别占总对数的42.7%、43.3%和14.0%,显著正关联种对数为1对,极显著和显著负关联种对数为2对;Ochiai(OI)指数显示66对种对的OI值落在[0.6,1],反映出热带海岸次生林主要种对间关联程度较为紧密。综上所述,热带海岸次生林群落优势种群种对间种间关联明显,物种之间存在一定的独立性,整体趋向稳定发展,实践中为加快热带次生林演替的进程,应重点保护正联结性较强的树种。     

重庆市不同规模灌区农灌水有效利用系数测算与对比研究

通过对重庆市不同规模灌区农田灌溉用水有效利用系数计算结果的比较研究,对出现的中型灌区农田灌溉用水有效利用系数大于小型灌区的结果进行合理性分析,并为重庆市农田灌溉用水有效利用系数的进一步提高,提出相关建议,为政府相关部门的科学决策提供参考.综合考虑重庆的地形、气候、土壤等因素.根据选定灌区的实测数据,采用首尾测算法计算2017年重庆市不同规模灌溉农田灌溉水的有效利用系数,采用单因素和主成分分析法,分析影响农田灌溉水有效灌溉系数的主要因素. 1)通过测算, 2017年重庆市中型灌区农田灌溉水有效利用系数为0.499,小型灌区为0.487,推算全市为0.495; 2)在影响农田灌溉水有效利用系数的主要因素中,当地的节水工程和管理状况所占的比例大于自然因素; 3)中型灌区在节水工程的投资、管理和维护等方面均优于小型灌区; 4)自然因素中,降水量对农田灌溉水有效利用系数具有一定的负面影响.     

Differential Cl−/Salt Tolerance and NaCl‐Induced Alternations of Tissue and Cellular Ion Fluxes in Glycine max,Glycine soja and their Hybrid Seedlings

The salt‐sensitive Glycine max N23674 cultivar, the salt‐born Glycine soja BB52 population, and their hybrid 4076 strain (F₅) selected for salt tolerance generation by generation were used as the experimental materials in this study. First, the effects of NaCl stress on seed germination, tissue damage, and time‐course ionic absorption and transportation were compared. When qualitatively compared with seed germination appearance in culture dishes, and tissue damages on roots or leaves of seedlings, or quantitatively compared with the relative salt injury rate, the inhibition on N23674 was all the most remarkable. After the exposure of 140 mm NaCl for 1 h, 4 h, 8 h, 12 h, 2 days and 4 days, the content of Cl^? gradually increased in the roots and leaves of seedlings of BB52, 4076 and 23674. Interestingly, the extents of the Cl^? rise in roots of the three experimental soybean materials were BB52 > 4076 > N23674, whereas those in leaves were just on the contrary. Secondly, by using the scanning ion‐selective electrode technique (SIET), fluxes of Na⁺ and Cl^? in roots and protoplasts isolated from roots and leaves were also investigated among the three experimental soybean materials. After 140 mm NaCl stress for 2, 4 and 6 days, and when compared with N23674, slighter net Cl^? influxes were observed in root tissue and protoplasts of roots and leaves of BB52 and 4076 seedlings, especially at the cellular protoplast level. The results indicate that with regard to the ionic effect of NaCl stress, Cl^? was the main determinant salt ion for salt tolerance in G. soja, G. max and their hybrid, and the difference in their Cl^?/salt tolerance is mainly attributed to the capacity of Cl^? restriction to the plant above‐ground parts such as leaves.     

Genomewide association study reveals novel quantitative trait loci associated with resistance towards Septoria tritici blotch in North European winter wheat

Fungal diseases are a major constraint for wheat production. Effective disease resistance is essential for ensuring a high production quality and yield. One of the most severe fungal diseases of wheat is Septoria tritici blotch (STB), which influences wheat production across the world. In this study, genomewide association mapping was used to identify new chromosomal regions on the wheat genome conferring effective resistance towards STB. A winter wheat population of 164 North European varieties and breeding lines was genotyped with 15K single nucleotide polymorphism (SNP) wheat array. The varieties were evaluated for STB in field trials at three locations in Denmark and across 3 years. The association analysis revealed four quantitative trait loci, on chromosomes 1B, 2A, 5D and 7A, highly associated with STB resistance. By comparing varieties containing several quantitative trait loci (QTL) with varieties containing none of the found QTL, a significant difference was found in the mean disease score. This indicates that an effective resistance can be obtained by pyramiding several QTL.     

Improvements of TKC Technology Accelerate Isolation of Transgene-Free CRISPR/Cas9-Edited Rice Plants

Elimination of the CRISPR/Cas9 constructs in edited plants is a prerequisite for assessing genetic stability, conducting phenotypic characterization, and applying for commercialization of the plants. However, removal of the CRISPR/Cas9 transgenes by genetic segregation and by backcross is laborious and time consuming. We previously reported the development of the transgene killer CRISPR (TKC) technology that uses a pair of suicide genes to trigger self-elimination of the transgenes without compromising gene editing efficiency. The TKC technology enables isolation of transgene-free CRISPR-edited plants within a single generation, greatly accelerating crop improvements. Here, we presented two new TKC vectors that show great efficiency in both editing the target gene and in undergoing self-elimination of the transgenes. The new vectors replaced the CaMV35S promoter used in our previous TKC vector with two rice promoters to drive one of the suicide genes, providing advantages over our previous TKC vector under certain conditions. The vectors reported here offered more options and flexibility to conduct gene editing experiments in rice.     

Pedotransfer functions to assess adsorbed phosphate using iron oxide content and magnetic susceptibility in an Oxisol

Adsorbed phosphate in soils can be chemically extracted; however, this process is both time‐consuming and not cost‐effective if large numbers of samples have to be analysed. Indirect assessment of adsorbed phosphate by pedotransfer functions (PTFs) can help optimize fertilizer strategies. This study aimed to evaluate the spatial variability of adsorbed phosphate (P_ads), iron oxides and magnetic susceptibility (MS) in oxisols and to calibrate PTFs to predict P_ads. A total of 308 soil samples were collected from Hapludox and Eutrudox soils formed from sandstone in Brazil. The contents of clay (196–607 g/kg), iron oxides (40–165 g/kg), MS (1.2–29 × 10^?6 m³/kg) and P_ads (327–842 mg/kg) were in the range of typical values for these highly weathered soils. This study showed that the attributes studied were spatially dependent. Geomorphic surfaces enabled understanding of spatial variability and helped to develop a more efficient sampling scheme to calibrate PTFs. Moreover, the adsorbed phosphate in these oxisols could be predicted by a PTF using iron oxides and MS as predictors. The MS attribute enabled the most accurate prediction (concordance coefficient = 0.95, root‐mean‐square error = 46 mg/kg and relative improvement in root‐mean‐square error = ?4.12) of spatial variability through PTF compared to other predictors.     

Abscisic Acid May Alter the Salicylic Acid‐Related Abiotic Stress Response in Maize

The effect of 0, 0.05 or 0.1 mm abscisic acid treatment on chilling tolerance and salicylic acid‐related responses was investigated in young maize seedlings (Zea mays L., hybrid Norma). Although the pre‐treatment of maize seedlings with abscisic acid slightly decreased the chlorophyll content, it also reduced the level of chilling injury caused by 6 days of cold treatment at 5 °C. Under normal growth conditions, increased levels of bound salicylic acid and of bound ortho‐hydroxycinnamic acid were observed in the leaves during abscisic acid treatment. In the roots, abscisic acid did not affect the free and bound salicylic acid levels, but increased the amount of free and bound ortho‐hydroxycinnamic acid. The activity of glutathione‐S‐transferase increased on the 3rd day of abscisic acid treatment, whereas it did not change when followed by cold stress, compared with the control leaves. In the roots, the activities of glutathione reductase, glutathione‐S‐transferase and ascorbate peroxidase increased during the abscisic acid treatment, and those of glutathione‐S‐transferase and ascorbate peroxidase were also stimulated when abscisic acid pre‐treatment was followed by cold stress, compared with the control roots. Our results suggest that an overlap may exist between the abscisic acid‐induced cold acclimation and the salicylic acid‐related stress response.     

加州鲈弹状病毒三水株全基因组序列测定及分析

【目的】对加州鲈弹状病毒弱毒三水株(MSRV-SS-7)及其母源株(MSRV-SS)进行全基因组克隆和测序分析。【方法】采用分段扩增方法对MSRV-SS-7及MSRV-SS基因组核心序列进行PCR扩增,第1轮根据鳜鱼弹状病毒(SCRV)基因序列(NC_008514.1)设计10对引物进行扩增并测序;在第1轮扩增产物测序、拼接基础上,设计8对引物进行第2轮扩增,拼接后获得基因组核心序列。同时用cDNA末端快速扩增法(RACE)获得3′和5′末端序列。采用Vector NTI 8.0对基因组核心序列和末端序列进行拼接,获得基因组全序列,使用Clone Manager 8.0对MSRV-SS-7和MSRV-SS进行序列比对分析;同时使用DNAMAN将基因组G基因序列转换为氨基酸序列,并通过MEGA 5.0与其他鱼类弹状病毒的G蛋白氨基酸序列进行进化树分析。【结果】MSRV-SS-7及其母源株MSRV-SS基因组全长均为11 548 bp,包含N、P、M、G、L等5个结构基因,基因组结构为3′Leader-N-P-M-G-L-Trailer 5′;MSRV-SS-7与MSRV-SS全基因组中共有10处核苷酸不同。G蛋白进化分析表明,MSRV-SS-7及母源株MSRV-SS与SCRV亲缘关系最近,且与鲈鱼弹状病毒属(Perhabdovirus)聚为一支。【结论】克隆分析了MSRV-SS-7的全基因组序列,可以用于后续活疫苗的开发。