水稻育性调控相关基因RNAi载体的构建及其表型鉴定

利用RNAi干扰技术研究不同基因对花粉发育、卵细胞发育和合子胚发育的影响是一种重要的手段。本研究通过筛选水稻在生殖发育过程中的9个重要调控基因,构建基因的RNAi表达载体,分析转基因植株育性及相关性状表型,以期探明RNAi表达载体对靶标性状的干扰效应。其中,AT61~AT63、AT64~AT66、AT67~AT69表达载体分别靶标花粉育性、卵细胞发育以及合子胚发育的调控。结果表明,除RNAi表达载体AT64没有获得转基因植株外,其余8个RNAi表达载体均获得了转基因植株;对T0代转基因植株的花粉育性、结实率以及潮霉素筛选(40 mg/L)发芽率检测的统计结果显示,RNAi表达载体AT62(花粉发育调控)、AT65(卵细胞发育调控)和AT67(合子胚发育调控)的干扰效应较强。本研究结果将为创制新型水稻基因工程不育系提供策略和选择。     

Effects of main cereal type and feed form on production performance,egg quality and egg sanitary indices of laying hens

ABSTRACT

1. The experiment was conducted to evaluate the effects of cereal types (maize or wheat) and feed forms (pelleted or mash feed) on production performance, egg quality and egg sanitary indices in laying hens.

2. Three hundred and sixty hens (Jinghong No. 1) at 18 weeks of age were randomly assigned to four treatments with six replicates of 15 hens per replicate according to a 2 × 2 factorial design with two cereal types (maize or wheat) and two feed forms (pelleted or mash feed).

3. Compared with the wheat-based diet, the maize-based diet improved (P < 0.05) average egg weight of laying hens. Yolk colour of hens fed with the maize-based diet was higher (P < 0.05) in comparison to those fed the wheat-based diet, while Haugh units were lower (P < 0.05) for the maize-based treatment. Egg mass and average daily feed intake of hens fed the pelleted diet were higher (P < 0.05) than of those fed the mash diet. However, the mash diet improved (P < 0.05) yolk colour compared with the pelleted diet. The percentage of dirty eggs for the wheat-based diet was higher (P < 0.05) than for the maize-based diet. The percentage of dirty eggs was higher (P < 0.05) in birds fed the pelleted diet compared with those fed the mash diet. There were interactions (P < 0.05) between cereal type and feed form, with regard to average egg weight and shell thickness.

4. In conclusion, dietary cereal type affected average egg weight, yolk colour and Haugh units in eggs, while feed form influenced egg mass, average daily feed intake and yolk colour.     

Identification of potential genomic regions for egg weight by a haplotype-based genome-wide association study using Bayesian methods

ABSTRACT

1. Theoretically, haplotype blocks might be a more suitable alternative to SNP genotypes as they are usually better at capturing multi-allelic QTL effects, compared to individual SNP genotypes in genome-wide association studies. The objectives of this study were to identify genomic regions related to egg weight traits by Bayesian methods (BayesA, BayesB, and BayesN) that fit fixed-length haplotypes using GenSel software.

2. Genotypes at 294,705 SNPs, that were common on a 600K Affymetrix chip, were phased for an egg-laying hen population of 1,063 birds. Recorded traits included first egg weight (FEW) and average egg weight at 28, 36, 56, 66, 72 and 80 weeks of age.

2. Fitting 1Mb haplotypes from BayesB resulted in the highest proportion of genetic variance explained for the egg weight traits. Based on the trait, the genetic variance explained by each marker ranged from 27% to 76%.

3. Different haplotype windows associated with egg weight traits only explained a small percentage of the genetic variance.

4. The top one 1-Mb window on GGA1 explained approximately 4.05% of total genetic variance for the FEW. Candidate genes, including PRKAR2B, HMGA2, LEMD3, GRIP1, EHBP1, MAP3K7, and MYH were identified for egg weight traits.

5. Several genomic regions, potentially associated with egg weight traits, were identified, some of which overlapped with known genes and previously reported QTL regions for egg production traits.     

The Effects of an Oxygen Scavenger and Coconut Water on Equine Sperm Cryopreservation

Alternative sources of lipoproteins in semen extenders could replace animal by-products. We hypothesized that: (1) post-thaw semen parameters and fertility would not be different in coconut water (CW)–treated samples compared with egg yolk (EY)–treated samples and (2) the use of an oxygen scavenger (Oxyrase) would improve post-thaw sperm motility and membrane integrity and decrease lipid peroxidation. Experiment 1: three ejaculates each from five stallions were split into four treatments: EY, CW, egg yolk with Oxyrase, and coconut water with Oxyrase. Computer-assisted sperm analysis measured progressive and total motility, velocity, and linearity. Membrane integrity, apoptosis, and lipid peroxidation were evaluated using propidium iodide, annexin, and BODIPY fluorescent probes, respectively. Samples were cryopreserved, stored in liquid nitrogen, and then thawed to 37°C and analyzed again. Experiment 2: one ejaculate was divided into two aliquots and cryopreserved using either CW or EY. In a crossover design, 12 mares were bred on two consecutive cycles with either EY or CW. Pregnancy evaluations were at 14-day gestation. No differences were detected in sperm parameters between CW and EY (P > .05). Oxyrase did not improve sperm motility parameters in post-thaw samples, nor did it show protective effects for viability or against membrane damage (P > .05). More mares became pregnant using CW than EY (11/12 vs. 6/12, respectively; P = .013). Use of CW is a viable alternative to animal-based products in the cryopreservation of stallion semen.     

黑龙江省大豆胞囊线虫种群分布和卵密度研究

对采自黑龙江省35个市县112份大豆田土样,采用蔗糖梯度离心法,研究了大豆胞囊线虫种群分布和种群密度.结果表明:从黑龙江省采集的所有大豆根围土样都检测到了大豆胞囊线虫,各地胞囊线虫的种群密度不同,每100 g干土中的虫卵量差别大,100 g干土中,嫩江县的大豆胞囊线虫卵量多达30 583个,最少的是抚远县,100 g干...     

饲料中添加不同水平的维生素E对半滑舌鳎(Cynoglossus semilaevis)亲鱼繁殖性能及后代质量的影响

在饲料中分别添加0、200、1200 mg/kg的维生素E(DL-α-生育酚乙酸脂形式)配制成Ec、El和Eh三种试验饲料,饲喂3龄半滑舌鳎亲鱼(雌性1.58±0.19 kg/尾、雄性0.20±0.05 kg/尾)70 d,探讨饲料中添加不同水平的维生素E对半滑舌鳎亲鱼繁殖性能的影响。结果显示,饲料中添加1200 mg/kg的维生素E(Eh组)不仅可以促进亲鱼性腺的发育,提高亲鱼产卵量、浮卵率、受精率、孵化率以及初孵仔鱼长度,而且影响仔鱼质量,仔鱼的成活率、12 d仔鱼长度、生存活力等方面均要高于对照组。而饲料中添加200 mg/kg的维生素E(El组)只对亲鱼性腺的发育和初孵仔鱼长度有一定的促进作用,对亲鱼产卵量、浮卵率、受精率、孵化率没有显著影响(P>0.05)。随饲料中维生素E的增加,血清中SOD的活性显著升高(P<0.05),MDA含量则显著下降(P<0.05)。此外,El和Eh试验组卵中干物质、维生素C的含量,以及卵子脂肪酸中PUFA、n-3PUFA、n-3HUFA和n-6PUFA的含量均要高于Ec组,El和Eh试验组之间的没有显著性差异(P>0.05)。结果表明,在半滑舌鳎亲鱼饲料中添加适量维生素E会亲鱼促进性腺的发育,增加产卵量,提高卵子和仔鱼的质量。在本试验中Eh组(1200 mg/kg)亲鱼的繁殖性能要优于Ec组(0 mg/kg)和El组(200 mg/kg)。     

徐海鸡蛋品质性状间的典型相关分析

为研究徐海鸡蛋品质性状间的内在关系,随机测定40周龄徐海鸡蛋品质,并运用典型相关分析对蛋内部品质性状和蛋壳品质性状的12个变量进行分析。结果表明:蛋内部品质中蛋重、蛋黄重、蛋白高度和哈氏单位与大部分蛋内部品质性状呈显著或极显著相关;除蛋形指数外其余部分蛋壳品质性状间呈显著或极显著相关。典型相关分析结果表明徐海鸡蛋内部品质性状和蛋壳品质性状的第1个典型相关达到极显著水平(P0.01),相关系数为0.8552,其相关信息占两组性状间总信息的79.67%,其中起主要作用的性状包括蛋白高度和蛋壳厚度。该研究结果对徐海鸡的选育利用具有指导意义。     

重组减蛋综合征病毒KnobS干酪乳杆菌的构建与表达

以干酪乳杆菌为传递载体,以开发食品级安全的减蛋综合征病毒疫苗为目标,构建了一个温度敏感型自杀型质粒系统p ORZP-UKD。将该质粒电转化到干酪乳杆菌L.casei中,经过2次升温处理和5-氟尿嘧啶抗性筛选,挑选阳性克隆,采用PCR和SDS-PAGE方法进行鉴定。KnobS基因整合到干酪乳酸杆菌基因组内,并实现融合蛋白KnobS的分泌表达。表明得到了一株具有无任何选择性标记、携带有KnobS表达盒元件的基因组整合的重组干酪乳酸杆菌KnobSΔupp L.casei,且整合的外源基因能够随着细菌的复制而进行表达,为减蛋综合征病毒免疫保护性抗原KnobS疫苗的进一步研制提供了试验数据。     

Is spawning stock biomass a robust proxy for reproductive potential?

Reference points based on fishing mortality (F) and spawning stock biomass (SSB) are a requirement of many fisheries management frameworks. SSB is assumed to be a proxy for stock reproductive potential (SRP). Limit reference points based on SSB are used to indicate the level of biomass below which productivity is affected. SSB fails to account for changes in fecundity, egg viability and sex ratio, and it has been argued that total egg production (TEP) provides a better reflection of SRP. We explore how accounting for TEP impacts limit reference points and evidence for a relationship between stock and recruit. Time series of SSB and TEP are compared for three North Sea stocks: cod (Gadus morhua), herring (Clupea harengus) and plaice (Pleuronectes platessa). Dynamics based on TEP are different from those based on SSB for cod and plaice, but the stock–recruit relationships were not ‘improved’ using TEP. Shifts in productivity (spawner per recruit) occur in all three time series and SSB underestimated uncertainty. Yet again, it was shown that assumptions of stationarity about fish population productivity are incorrect. We argue that the use of TEP does improve the realism in our understanding of stock dynamics, and demographically, more complex management strategy evaluation is required to develop management procedures that are robust to uncertainty and integrate F and the demographic health of a stock. Empirical feedback control systems based on fisheries independent indices including surveys of eggs, larvae, recruits, juveniles or spawning adults should be evaluated and compared to traditional approaches.     

基于多信息融合的疫苗制备中鸡蛋胚体分拣系统

采用多信息融合方法研究疫苗制备中鸡蛋胚体状态的识别与分拣技术。在研究不同状态鸡蛋胚体图像特征、温度衰减和透光度变化情况的基础上,得出活胚图像中血管多、粗且呈放射状,弱胚图像中血管少、细且断裂,死胚图像内部均匀无血管,污染胚图像内部有明显黑块特征;鸡蛋胚体从37.8℃的孵化箱中取出置于25℃室温环境,活胚、弱胚、污染胚、死胚的温度衰减速度依次增大;活胚的透光度随孵化时间增加而逐渐降低,其他胚体透光度变化相对较小。将图像、温度、透光度信息特征融合,建立BP神经网络信息融合模型对鸡蛋胚体状态进行识别。最后,从37.8℃孵化箱中抽取80枚孵化6 d的鸡蛋胚体放置于室温10 min后,采集图像、温度和透光度信息,进行试验验证。结果表明多信息融合系统的识别准确率为96.25%,比单用图像、温度和透光度传感器进行识别的准确率分别提高了6.25%、13.75%和8.75%。