CRISPR/cas系统及其在家禽上的应用研究进展

CRISPR/cas是一种获得性免疫防御系统,广泛存在于细菌和古细菌中。Strep tococcus pyogenes cas9(Spcas9)作为目前研究最多、最清楚的效应蛋白,因其酶活性高和靶点广而被应用于生命科学各个领域,但Spcas9分子量大、脱靶率高等缺点在一定程度上限制了其应用。随着CRISPR系统技术被深入开发,一些新效应蛋白被发现,本文就CRISPR/cas系统分类、原理、新效应蛋白发现及cas9(S.Pyogenes)多种变构体技术开发在及家禽上的应用进行综述,为相关领域的研究提供参考。     

不同水肥浓度对涌泉根灌枣树叶面积和产量的影响

以陕西榆林山地矮化密植梨枣树为研究对象,分析了不同水肥浓度对涌泉根灌枣树生育期内叶面积指数、果实发育和产量的影响。结果表明,不同水肥处理叶面积指数在枣树果实膨大期达到高峰,且稳定持续至果实成熟期末期,随后叶面积指数开始下降。叶面积指数随着萌芽展叶期灌水量的增加而增加,所有水肥处理下,枣数叶面积指数最大的是高水中肥处理(W1N2)；所有处理中,高水、低水处理与低肥相组合,果实体积最大,中灌水处理为不施肥时果实的体积最大。单位枝长的果实数目最高的是中水中肥处理(W2N2),最低的是低水处理(W3),在一定范围内,适当施肥能促进枣果单位枝长的果实数目的增加,但肥液浓度过大,反而会抑制其增加。在一定条件下,所有处理中产量最高的是中水中肥处理(W2N2),产量超过2000kg/亩,最低的是低水高肥处理(W3N1),产量不到1000kg/亩。因此可以选择中水中肥处理(W2N2),能获得最高的产量。     

Effects of taurine-zinc on learning and memory abilities of vascular dementia mice

AIM:To observe the effects of taurine-zinc (TZC) on the learning and memory abilities of vascular dementia (VD) mice and to investigate the related mechanism. METHODS:The mice were randomly divided into model group, sham group, and TZC at 50 mg/kg, 100 mg/kg and 200 mg/kg groups. The mice in drug groups were given TZC by gavage at 10 mL/kg once daily. The mice in sham group and model group were given equal volume of distilled water. VD mice were established by intercepting both common carotid arteries and bleeding at caudal vein after 14 d of gavage. The levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were detected by ELISA. The levels of inducible nitric oxide synthase (iNOS) and nitric oxide (NO) were measured via spectrophotometer. Step-down test and Morris water maze test were used to examine the abilities of learning and memory in the mice. RESULTS:TZC at 50 mg/kg, 100 mg/kg and 200 mg/kg reduced the levels of TNF-α, IL-1β, iNOS and NO in the brain tissues. In the water maze test, TZC at 100 mg/kg and 200 mg/kg significantly decreased the error times and latency compared with model group. In the step-down test, the escape latency was prolonged and error times were lowered significantly by treatment with TZC at 50 mg/kg, 100 mg/kg and 200 mg/kg as compared with model group. CONCLUSION:TZC improves the abilities of learning and memory, which might be related to the reduction of TNF-α, IL-1β, iNOS and NO levels in VD mice.     

Zerumbone attenuates MPP+-induced cytotoxicity in human neuroblastoma SH-SY5Y cells by inhibition of oxidative stress

AIM: To investigate the role of zerumbone (ZER) in 1-methyl-4-phenylpyridinium (MPP⁺)-induced cytotoxicity of human neuroblastoma SH-SY5Y cells. METHODS: Human neuroblastoma SH-SY5Y cells were cultured in vitro and the protective effect of ZER against MPP⁺-induced cytotoxicity was measured by CCK-8 assay. Flow cytometry was used to determine the apoptosis and reactive oxygen species (ROS). The expression of Parkinson disease protein 7 (PARK7) was knocked-down by using PARK7-specific short hairpin RNA (shRNA). The protein levels of PARK7, nuclear factor E2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) were determined by Western blot. RESULTS: MMP⁺ remarkably reduced the cell viability in a dose-dependent and time-dependent manner. The SH-SY5Y cell injury model was established by treatment with MPP⁺ at 600 μmol/L for 24 h. ZER up-regulated the protein levels of PARK7 and Nrf2 (P<0.05), alleviated apoptosis (P<0.05), and reduced ROS production (P<0.05) in the SH-SY5Y cell injury model. Meanwhile, N-acetyl-L-cysteine (NAC) had the similar functions. Moreover, significant reductions in the protein levels of Nrf2 and HO-1 (P<0.05), and obvious increases in apoptosis (P<0.05) and ROS level (P<0.05) were demonstrated in PARK7-knockdown cells. CONCLUSION: ZER protects SH-SY5Y cells against MPP⁺-induced cytotoxi-city, which may be related to activation of PARK7/Nrf2/HO-1 pathway, and subsequent attenuation of oxidative stress and apoptosis.     

果蔬型甜玉米新品种京科甜191的选育

京科甜191是以超甜玉米自交系T68为母本,自交系T6302为父本,通过杂交选育而成的果蔬型甜玉米品种。果穗筒型,穗长19.0 cm,穗粗5.3 cm,穗行数16~18行,行粒数36~38粒,粒行整齐。籽粒亮黄色,外观漂亮。平均每667 m2鲜果穗产量900 kg左右,出籽率约67.4%。鲜籽粒含粗蛋白2.98%,粗脂肪1.37%,总糖6.7%,口感甜脆清爽,皮薄无渣。播种至采收平均85.4 d(天),适宜京津冀及类似生态区种植。     

Assessing vulnerability of New Zealand lakes to loss of conservation value from invasive fish impacts

1. Predictions of invasion risk for seven non‐indigenous fish species, ecological impact scores for individual species, and lake conservation rankings were linked to develop Invasion Risk Impact (IRI) and Lake Vulnerability (LV) indices that help identify New Zealand lakes most at risk of loss of conservation value from potential multi‐species invasions.
2. Species‐specific IRI scores (the product of predicted invasion risk and species impact) highlighted Eurasian perch (Perca fluviatilis) and the brown bullhead (Ameiurus nebulosus), as the species most likely to spread and cause ecological harm in lakes. For 3431 lakes >1 ha throughout New Zealand, total IRI tended to be highest for lowland riverine and dune lakes most of which are already colonized by multiple invasive fish species.
3. The LV index indicated that lakes most at risk of loss of conservation value from invasive fish impacts were predominantly (i) in the northern half of the North Island where several uncommon lake types occur, and (ii) along the west coast of the South Island where conservation value is often greater, largely because of low catchment modification.
4. The IRI and LV indices can be used to assist with setting priorities for surveillance monitoring, advocacy, and response planning targeted at preventing the establishment of invasive fish in moderate‐to‐high value lakes most susceptible to ecological impacts. Both indices can be adapted to accommodate alternative impact and conservation scoring systems, providing a flexible tool for local‐ and national‐scale assessments of lake vulnerability to fish invasion impacts.

Copyright © 2016 John Wiley & Sons, Ltd.     

华南6水系与澜沧江-湄公河攀鲈线粒体ND2基因的遗传多样性分析

测定了中国华南6水系及澜沧江(云南勐腊)-湄公河流域(柬埔寨洞里萨湖)的125尾攀鲈(Anabas testudineus)线粒体部分ND2基因1 010 bp序列,分析发现39个变异位点和12个单倍型,总遗传多样性较低(h=0.369,π=0.003 8),推测可能经历过严重的瓶颈效应;中国攀鲈群体遗传多样性更低(h=0.282,π=0.000 4),处于边缘区的中国攀鲈群体是造成低遗传多样性的主要原因。在单倍型网络图中柬埔寨和中国攀鲈各自聚类,具有明显地理结构和谱系结构,推测地质运动和气候变化导致基因交流受阻所致。核苷酸错配图和中性检验表明中国群体经历过种群扩张,时间约为(5.94~4.13)万年前。华南水系群体间基因交流通畅,不存在明显分化;但与云南澜沧江群体间分化大而显著(FST=0.775,P0.01),AMOVA分析显示变异主要来自组群间(77.41%),推测二者分化时间约为(4.0~2.8)万年前,云南群体受末次冰期的影响,基因交流受阻而出现分化。中国群体和柬埔寨群体可作为2个管理单位进行保护;就中国群体而言,韩江水系群体遗传多样性最高,建议优先保护;澜沧江与华南水系间群体分化显著且遗传多样性极低,建议对澜沧江水系群体进行保护,以避免种质资源灭绝。     

Effects of dietary protein and lipid levels on growth performance,fatty acid composition and antioxidant‐related gene expressions in juvenile loach Misgurnus anguillicaudatus

Dojo loach (Misgurnus anguillicaudatus) is one of the most important cultured freshwater fish in several East Asian countries. However, a little information is available in its nutritional requirements. Thus, this study was conducted to evaluate the effects of feeding varying levels of dietary protein and lipid on growth, fatty acid composition and antioxidant‐related gene expressions in juvenile loach. Six practical diets at three levels of protein (30%, 40% and 50%) and two levels of lipid (6% and 12%) were fed to loach juveniles (initial weight 0.40 g) in triplicated groups (20 fish per replicated) for a period of 8 weeks. Results showed that regardless of lipid level, body weight gain of fish was significantly increased with incremental dietary protein level. Meanwhile, feed conversion ratio was significantly decreased by dietary protein levels, and the lowest value was observed in fish fed dietary protein levels of 50%, regardless of dietary lipid level. Moreover, the percentage of 22:6n‐3 in viscera was significantly increased by different protein levels. The expression level of catalase was significantly increased with incremental dietary protein level with both lipid levels. Meanwhile, the expression level of hepatic nuclear factor erythroid 2‐related factor 2 (Nrf2) was downregulated with incremental dietary protein level with 6% of lipid level, but the expression was upregulated with incremental dietary protein level with 12% of lipid level. In conclusion, these data suggested that 6% lipid and 50% protein in diet was optimal for loach during early development stage.     

Sorghum extrusion process combined with biofortified sweet potato contributed for high iron bioavailability in Wistar rats

This study aimed to evaluate the effect of sorghum and sweet potato on the bioavailability of iron, gene expression of proteins involved in iron metabolism and the plasma antioxidant capacity in animals fed with whole sorghum grains processed by dry heat or extrusion, combined or not with sweet potato flour with high content of carotenoids. Five experimental groups were tested (n = 7): dry heat sorghum flour (DS); extruded sorghum flour (ES); whole sorghum flour + sweet potato flour (DS + SP); extruded sorghum flour + sweet potato flour (ES + SP) and positive control (FS). The evaluations included: hemoglobin gain, hemoglobin regeneration efficiency, gene expression of divalente metal transporter 1 (DMT-1), duodenal citochroma B (DcytB), ferroportin, hephaestin, transferrin and ferritin and total plasma antioxidant capacity (TAC). The ES + SP group showed higher (p < 0.05) expression of DcytB, ferroportin and hephaestin when compared to the control group. The DS group showed high (p < 0.05) expression of DMT-1 and the ES showed high mRNA expression of transferrin and ferritin. The changes in the sorghum physicochemical properties from extrusion process reduced the iron and phytate content, and increased the gene expression of proteins involved in iron metabolism, improving iron bioavailability. The combination of sweet potato and sorghum flour (dry or extruded) improved the iron capture and total antioxidant capacity, probably due to the presence of β-carotene and antioxidant compounds.     

Characterizing the pigmented traditional rice cultivars grown in temperate regions of Kashmir (India) for free and bound phenolics compounds and in vitro antioxidant properties

The purpose of the research was to identify the phenolic and flavonoid compounds of seven different traditional pigmented whole rice cultivars grown in the temperate regions of Kashmir so as to study their relationship with in vitro antioxidant capacities. The completely pigmented rice cultivars were found to have higher phenolic, flavonoid, anthocyanin contents and exhibited higher antioxidant capacities than the light colored and sparely colored rice cultivars. A total of 40 compounds had been identified in the analyzed rice cultivars that were found to be distributed in 6 major categories with 6-phenolics, 6-flavonoids, 11-hydroxycinnamic acid derivatives, 7-hydroxybenzoic acid derivatives, 3-anthocyanins and 7-flavonoid glucosides of different flavonoid compounds. Among the free and bound fractions for each cultivars the light and sparsely colored depicted higher content of phenolics and in vitro antioxidant properties in bound faction, while the completely pigmented cultivars showed higher antioxidant properties in free fractions. The anthocyanins quercetin-3-O-galactoside, cyanidin-3-O-rutinoside and pelargonidin-3-O-diglucoside had been identified by LC-MS existing in the free fractions of the analyzed rice cultivars whereas, the free fraction of acetone + H₂O possessed higher percentage of phenolic compounds as compared to methanolic extracts and bound fractions. The black colored cultivars possessed higher DPPH scavenging activity and lipid peroxidation inhibition.