山羊TNNT3基因可变剪切及其对骨骼肌细胞分化的作用

【目的】快速骨骼肌肌钙蛋白T（fast skeletal troponin T3,TNNT3）作为肌钙蛋白(troponin, Tn)家族成员,调节横纹肌收缩、参与骨骼肌的生长发育并影响家畜肉质性状。通过获得山羊TNNT3基因的可变剪切体,分析山羊TNNT3基因可变剪切的表达模式及其在肌细胞分化中的作用,深入解析TNNT3基因在山羊骨骼肌生长发育过程中的作用机制。【方法】基于NCBI已公布山羊TNNT3基因（NM_001314210.1）和牛TNNT3基因（XM_010821200）mRNA序列,使用软件Primer Premier 6.0设计引物,以简州大耳羊胚胎期和出生后7个阶段骨骼肌为试验材料,克隆测序获得山羊TNNT3基因的CDS区可变剪切体,利用软件ORF Finder、EditSeq、DNAMAN、ClustalW和MEGA_X_10.1.8等对序列进行生物信息学分析;进一步设计实时荧光定量（real-time PCR,RT-qPCR）及半定量引物,研究TNNT3基因剪切体在7个不同组织（背最长肌（longissimus dorsi muscle,LD）、半膜肌（semimembranosus muscle,SM）、心、肝、脾、肺、肾）和7个发育阶段（胚胎期E75、E90、E105和出生后B3、B45、B150、B300）肌肉组织（背最长肌和半膜肌）中表达模式;此外,对转录本TNNT3_3进行体外编码能力检测确定其具有编码蛋白的能力,并在山羊骨骼肌卫星细胞（skeletal muscle satellite cells,MuSCs）中过表达,观察细胞形态变化以及检测标志基因的表达变化,研究其对山羊MuSCs分化的作用。【结果】①TNNT3（NM_001314210.1）CDS区全序列主要含有18个外显子,其中外显子16/17相互排斥,转录后单一表达。克隆发现山羊TNNT3基因 5个新转录本（TNNT3_1—5）,其外显子数分别是15、15、20、16、14。②生物信息学分析结果显示山羊TNNT3基因核苷酸序列和氨基酸序列与绵羊、牛、猪等哺乳动物具有很高的一致性,而与鱼类和爬行类动物的一致性较低,说明TNNT3基因序列在哺乳动物高度保守。③TNNT3 mRNA在背最长肌、半膜肌、心、肝、脾、肺、肾7个组织中都有表达,其中在骨骼肌中高度富集(P < 0.01),心脏及肺次之,其余组织中较低;TNNT3 mRNA在背最长肌和半膜肌中的表达始终处于一个动态变化中,胚胎期TNNT3在半膜肌的表达量高于背最长肌（P<0.05）;出生后则背最长肌中高于半膜肌（P<0.05）。④山羊TNNT3基因转录本TNNT3_3重复出现保守的外显子9—11（138bp）,体外翻译实验显示其可编码蛋白且蛋白大小与预期基本相符（37 kD）;相较于对照组,在山羊MuSCs中过表达该转录本使肌分化标志基因Myomaker、MyoG和MyH4 mRNA极显著升高(P < 0.01)。【结论】获得了山羊TNNT3基因具有完整CDS区5个新可变剪切体,TNNT3主要在肌肉组织（背最长肌和半膜肌）中高表达,在哺乳动物中高度保守且促进成肌分化。初步表明TNNT3基因在动物肌肉生长发育中具有重要的生物学功能。     

酒糟和醋糟营养成分差异及其瘤胃降解特性分析

本试验旨在评价酒糟、醋糟等非常规饲料资源的饲用价值，为酒糟醋糟资源应用于肉牛养殖提供数据参考。试验选用山西不同地区酒糟、醋糟试样5个，试样编号分别为白酒糟、啤酒糟、醋糟1、醋糟2、醋糟3，发酵原料分别为白酒糟（高粱）、啤酒糟（大麦+大米）、醋糟1（高粱+大麦）、醋糟2（大麦+玉米）、醋糟3（高粱+玉米）。通过实验室常规方法测定样品常规营养成分以及利用体外产气法和尼龙袋法评价其饲用价值，同时建立体外法与尼龙袋法测定干物质降解率的回归方程。结果表明：5个样品的干物质（DM）含量为23.25% ~ 42.96%，粗蛋白质（CP）含量分别为啤酒糟（31.84%）>白酒糟（14.58%）>醋糟3（14.40%）>醋糟1（12.21%）>醋糟2（8.46%）。醋糟1的中性洗涤纤维（NDF）、酸性洗涤纤维（ADF）含量均为最高，其次为醋糟2、醋糟3，白酒糟NDF含量最低。啤酒糟的72 h产气量（GP72）显著高于白酒糟（P < 0.05）。体外产气法和尼龙袋法测定的酒糟72 h干物质降解率（IVDMD72、DMD72）显著高于醋糟（P < 0.05）。5个试样的IVDMD72与DMD成正相关，随着降解时间增加其相关性逐渐增加。综上，酒糟的CP含量较高（14.58% ~ 31.84%），醋糟NDF、ADF含量相对较高（59.59% ~ 64.91%，38.78% ~ 54.78%）。结合GP、IVDMD及DMD结果，酒糟消化性能优于醋糟。以IVDMD建立估算DMD的回归公式为DMD72=-19.215+1.645IVDMD72（R2=0.912，P < 0.05）|以IVDMD建立估算DM有效降解率（ED）的回归公式为ED=-12.448+1.045IVDMD72（R2=0.929，P < 0.01）。 [关键词] 醋糟|酒糟|体外产气法|尼龙袋法     

Transport of cadmium from soil to grain in cereal crops: A review

Due to rapid urbanization and industrialization, many soils for crop production are contaminated by cadmium(Cd), a heavy metal highly toxic to many organisms. Cereal crops such as rice, wheat, maize, and barley are the primary dietary source of Cd for humans, and reducing Cd transfer from soil to their grains is therefore an important issue for food safety. During the last decade, great progress has been made in elucidating the molecular mechanisms of Cd transport, particularly in rice. Inter-and intraspecific variations in Cd accumulation have been observed in cereal crops. Transporters for Cd have been identified in rice and other cereal crops using genotypic differences in Cd accumulation and mutant approaches. These transporters belong to different transporter families and are involved in the uptake, vacuolar sequestration, root-to-shoot translocation, and distribution of Cd. Attempts have been made to reduce Cd accumulation in grains by manipulating these transporters through overexpression or knockout of the transporter genes, as well as through marker-assisted selection breeding based on genotypic differences in Cd accumulation in the grains. In this review, we describe recent progress on molecular mechanisms of Cd accumulation in cereal crops and compare different molecular strategies for minimizing Cd accumulation in grains.     

小麦秸秆压缩弯曲特性试验研究

为给秸秆圆捆打捆装置和其它打捆机构提供相应的力学参数和理论基础,根据麦茎秆的几何尺寸,并利用万能试验机控制夹具加载速度对不同含水率的麦秸秆进行弯曲、轴向压缩和径向压缩等力学特性试验,得出载荷-位移等曲线,并获得相关力学参数数据。利用Origin将数据直观地反映出来,再用MatLab对试验数据进行拟合,得出拟合方程。试验结果表明:麦秸秆压缩所需要的力大于弯曲所需的力;含水率和夹具加载速度在一定范围内时,无论是在麦秸秆的弯曲试验还是压缩试验中,载荷峰值的大小与加载速度和含水率都有关,因此在设计打捆压缩成型机构时需考虑含水率与其关键机构工作转速对打捆装置的影响。     

鸡皮刺螨实验室饲养与繁殖技术研究进展

鸡皮刺螨是一种吸血性外寄生虫,会引起鸡消瘦、贫血,并导致产蛋下降,造成巨大经济损失。为给科研提供可靠虫源,并对杀螨药物、疫苗以及其他防控手段进行效果评价,国内外学者进行了鸡皮刺螨实验室的人工饲养与繁殖技术研究。文章综述了国内外鸡皮刺螨实验室饲养与繁殖技术的研究进展,将其总结归纳为使用动物饲养繁殖技术与离体饲养繁殖技术两类,同时指出了各项技术在应用过程中存在的问题,为该技术的进一步完善与发展提供相关参考。     

Straw mulching with fertilizer nitrogen: An approach for improving crop yield,soil nutrients and enzyme activities

Field experiments were conducted to study soil properties, soil enzymes activities, water use efficiency (WUE) and crop productivity after six years of soya bean straw mulching in the semi‐arid conditions of China. The experiment included four treatments: CK (Control), N (240 kg N ha^‐1), H (soya bean straw mulching at half rate 700 kg ha^‐1 with 240 kg N ha^‐1) and F (soya bean straw mulching at full rate 1,400 kg ha^‐1 with 240 kg N ha^‐1). Soil organic carbon (SOC), soil labile organic carbon (LOC), soil available N (AN), available P (AP) and enzyme activities were analysed after wheat harvesting in 2016 and 2017. Results show that straw amounts had positive effects on the soil fertility indices being higher for treatment F. The SOC, LOC, AN, AP and enzyme activities (i.e. saccharase, urease and alkaline phosphatase) were in the order of F > H > N > CK. High wheat grain yield and WUE were observed for F treatment. A total of six years mulching along with 240 kg ha^‐1 nitrogen fertilizer application is sufficient for wheat yield stability and improving soil properties except urease activities in the semi‐arid condition of China. However, the straw mulching amount should be further studied with minimum nitrogen fertilizer for an environment‐friendly and effective approach for improving the soil biological properties with adequate crop production on a sustainable basis in the semi‐arid region of China.     

小麦TaeEF1β基因的克隆、同源性及表达分析

真核翻译延伸因子(eukaryotic translation elongation factor,eEFs)是一种重要的多功能调控蛋白,eEF1β是eEF1的组成部分,在蛋白质生物合成过程中发挥着重要的作用。本文通过RT-PCR扩增克隆小麦(Triticum aestivum L.)的eEF1β基因,并命名为TaeEF1β。氨基酸同源性分析发现,TaeEF1β具有高度保守性,且其保守结构域位于137~226 aa处。qRT-PCR结果表明,中国小麦花叶病毒(Chinese wheat mosaic virus,CWMV)侵染小麦植株后,可以诱导TaeEF1β基因转录水平的上调表达。另外,本文也进一步分析了TaeEF1β基因在小麦根、茎、叶的表达水平和CWMV侵染不同时间点的表达情况。     

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.