Cloning and expression analysis of innate immune genes from red sea bream to assess different susceptibility to megalocytivirus infection

As suggested by the Office International des Epizooties (OIE), fishes belonging to the genus Oplegnathus are more sensitive to megalocytivirus infection than other fish species including red sea bream (Pagrus major). To assess the roles of the innate immune response to these different susceptibilities, we cloned the genes encoding inflammatory factors including IL‐8 and COX‐2, and the antiviral factor like Mx from red sea bream for the first time and performed phylogenetic and structural analysis. Analysed expression levels of IL‐1β, IL‐8 and COX‐2 and the antiviral factor like Mx genes performed with in vivo challenge experiment showed no difference in inflammatory gene expression or respiratory burst activity between red sea bream and rock bream (Oplegnathus fasciatus). However, the Mx gene expression levels in red sea bream were markedly higher than those in rock bream, suggesting the importance of type I interferon (IFN)‐induced proteins, particularly Mx, during megalocytivirus infection, rather than inflammation‐related genes. The in vitro challenge experiments using embryonic primary cultures derived from both fish species showed no difference in cytopathic effects (CPE), viral replication profiles, and inflammatory and Mx gene expression pattern between the two fish species.     

Molecular cloning and functional characterization of the fatty acid delta 6 desaturase (FAD6) gene in the sea cucumber Apostichopus japonicus

The sea cucumber (Apostichopus japonicus), an important echinodermata, had high value in nutrition and medicine for its rich collagen, sulphated polysaccharide, glycosides and polyunsaturated fatty acids (PUFA). The cDNA of the fatty acid desaturase gene in A. japonicus (AJFAD6) was cloned and was found to encode a desaturase with delta 6 FAD activity. Sequence analysis indicated that AJFAD6 included an open reading frame of 1392 bp, encoding 463 amino acids. AJFAD6 has all the conserved motifs found in other members of the FAD6 family, including an N‐terminal cytochrome b5 domain and three histidine‐rich regions. qRT‐PCR showed that AJFAD6 was expressed in all tissues tested during juvenile development and was mainly expressed in the respiratory tree at 150 days after adherence (150 days) and in the intestine at 100 days. Furthermore, AJFAD6 mRNA was also detected in the analysed adult tissues, with higher expression in the intestine and testis. Functional characterization of AJFAD6 in a recombinant yeast, Pichia pastoris, showed that AJFAD6 could catalyse exogenous linoleic acid (LA) and α‐linolenic acid (ALA) to produce γ‐linoleic acid (GLA) and stearidonic acid (STA), respectively, at conversion rates of 11.1% for LA to GLA and 3.4% for ALA to STA. Our results suggested that the biosynthetic pathway of PUFA existed in the sea cucumber, but endogenous production of eicosapentaenoic acid, arachidonic acid and docosahexaenoic acid from either LA or ALA precursor appeared to be limited.     

Apparent digestibility coefficients of animal and plant feed ingredients for juvenile Pseudobagrus ussuriensis

A study was conducted to determine apparent digestibility coefficients (ADCs) of dry matter (DM), crude protein (CP), lipid, gross energy, phosphorus and amino acids (AAs) from Peruvian fish meal (PFM), native fish meal (NFM), meat bone meal (MBM), dried daphnia meal (DDM), soybean meal (SBM), full‐fat soybean (FSBM), cottonseed meals (CSM), rapeseed meal (RSM) and corn gluten meal (CGM) for Pseudobagrus ussuriensis (7.8 ± 0.06 g). Test diets were formulated to contain a 70:30 mixture of reference diet to test ingredient with chromic oxide (5 g kg^–1) serving as the inert marker. ADCs of DM ranged from 49.94% (CGM) to 87.11% (PFM), the highest ADCs of DM were observed in PFM and the lowest was found in CGM. ADCs of CP in the test ingredients ranged from 74.92% (CGM) to 93.41% (SBM). ADCs of crude lipid ranged from 51.80% (CGM) to 94.07% (PFM) for fish. ADCs of energy ranged from 56.42% (CGM) to 90.09% (PFM). ADCs of phosphorus ranged from 21.81% (CSM) to 65.70% (PFM). AA availability for the test ingredients followed a similar pattern to ADCs of CP, in which values for PFM, NFM and MBM were generally higher (p < .05) than those of other protein ingredients. Among all plant meals, the AA availability values in SBM were higher (p < .05) than those in CGM, CSM and RSM. The availability of most AAs in CGM was the lowest (p < .05) among all tested ingredients. In conclusion, PFM and NFM are good sources of available protein and AAs. When fishmeal supply is limited, MBM and DDM are substitutes for fishmeal reduction in P. ussuriensis diets; at the same time, SBM and FSBM are also very acceptable protein feedstuffs. However, CSM, RSM and CGM could not be used at high levels in P. ussuriensis diets because of their inferior digestibilities.     

江西一季籼粳杂交稻稻曲病防治药剂及次数研究

选择合适的防治药剂和防治次数是减轻稻曲病危害的关键。本研究以籼粳杂交稻甬优12为材料,比较了6种药剂和3种喷施次数对稻曲病的防控效果和产量性状差异。结果表明,在6种药剂中,阿米妙收处理的病穗率防效和病指防效最好,分别达到91.53%和96.63%,但与其他药剂无显著性差异。在防治次数上,爱苗和拿敌稳均为防治3次的处理病指防效最好,分别为81.73%和80.15%,显著高于防治次数3次以下的处理。阿米妙收处理的产量最高,达11 719.52 kg/hm~2,显著高于其他处理。     

Silymarin inhibits adipogenesis in the adipocytes in grass carp <Emphasis Type="Italic">Ctenopharyngodon idellus</Emphasis> in vitro and in vivo

In this study, two experiments were performed to explore the function of silymarin in adipogenesis in grass carp (Ctenopharyngodon idellus) using in vitro and in vivo models. In experiment 1, differentiated grass carp pre-adipocytes were treated with silymarin for 6 days. Treatment with 100 μg mL^?1silymarin (SM100 group) significantly reduced triglyceride accumulation at day 6. The adipogenic gene expression levels of PPARγ, C/EBPα, SREBP1c, FAS, SCD1, and LPL, and the protein expression level of PPARγ were significantly down-regulated in the SM100 group. Additionally, the SM100 group had significantly lower reactive oxygen species production and reduced glutathione contents compared with the control in vitro. In experiment 2, the juvenile grass carp (mean body weight= 27.4 ± 0.17 g) were fed six isonitrogenous and isocaloric diets in a factorial design containing 0, 100, or 200 mg kg^?1 silymarin (SM0, SM100, SM200) associated with either 4 or 8% lipid levels (low lipid, LL, and high lipid, HL, respectively) for 82 days. The results demonstrated that dietary silymarin supplementation significantly reduced the elevated intraperitoneal fat index in grass carp fed with high-lipid diets, and the gene expression of adipogenesis (PPARγ, FAS) when supplemented with dietary silymarin was notably lower than when no silymarin was supplemented under the high-lipid diets. Thus, our data suggest that silymarin suppressed lipid accumulation in grass carp both in vitro and in vivo, and the effect might be due to an influence on the expression of adipogenesis factors and ROS production partly associated with effects on antioxidant capability.     

大豆PAL2-3基因的克隆及转化研究

大豆异黄酮是苯丙氨酸代谢途径中合成的一类次级代谢产物,在苯丙氨酸代谢途径中,苯丙氨酸解氨酶(PAL)是关键酶和限速酶。本课题组前期研究发现PAL基因的相对表达量与大豆异黄酮含量具有明显的协同增减趋势,并且在PAL基因家族成员时空表达模式分析中发现,PAL2-3(XM_003542493)是PAL基因家族中相对表达量较高的主要表达成员之一。本试验首先通过克隆大豆中PAL2-3基因;然后构建pCAMBIA3301-GmPAL2-3植物过表达载体,将构建好的pCAMBIA3301-GmPAL2-3表达载体重组质粒转化到根癌农杆菌EHA105中;采用农杆菌介导的大豆子叶节转化体系获得转化植株,并对T1代转化植株进行PPT检测,外源标记基因Bar检测以及荧光定量PCR检测,对转基因阳性植株进行大豆籽粒异黄酮含量的测定。结果表明T_1代转基因植株中PAL2-3基因的表达量是对照的5.11~11.24倍,总异黄酮含量最高的(2 587.63μg·g~(-1))是对照(1 616.90μg·g~(-1))的1.6倍。因此在大豆中过量表达PAL2-3基因可以提高大豆籽粒中异黄酮含量。     

薏苡紫色柱头性状的遗传分析

为研究薏苡紫色柱头性状的遗传规律,利用4份材料做亲本构建4个杂交组合,观察并记录其F 1、F 2植株的性状特征和分离比例。结果显示,紫色柱头相对于白色为显性,各F 2群体植株的柱头颜色分离比皆符合3∶1比例,说明紫色柱头是由1对显性等位基因控制。     

基于季节特征的土壤退墒模型建立与率定

利用河南省南阳市2011—2015年12个墒情站的土壤水分监测资料,分析研究了无增墒情况下土壤含水量消退规律,构建出基于不同季节(夏季、春秋季与冬季)的土壤含水量与衰减系数的数学模型,运用规划求解法率定模型参数,并利用2016—2017年资料对模型进行检验。结果表明,不同季节土壤水分衰竭系数模型分别为α=1.023(1-ω~2/4353)~(1/2)(夏季)、α=1.013(1-ω~2/7005)~(1/2)(春秋季)与α=1.008(1-ω~2/9303)~(1/2)(冬季),所建立的数学模型适用于南阳市相应季节壤土与粘壤土小麦、水稻与休闲地土壤旱情预测,但不适用于夏季与春秋季砂土特别是休闲地砂土土壤墒情预测。