山羊ACSS2基因的克隆、序列分析及组织表达研究

本研究旨在对山羊乙酰辅酶A合成酶2（acetyl-CoA synthetase 2,ACSS2）基因进行克隆和生物信息学分析,并检测其在山羊不同泌乳时期乳腺组织中的表达量变化。以山羊乳腺组织RNA为模板,采用RT-PCR方法扩增并克隆山羊ACSS2基因完整CDS区序列,对测序结果进行生物信息学分析,并对ACSS2基因在山羊不同泌乳时期乳腺组织中的表达量进行分析。结果显示,山羊ACSS2基因CDS区序列长2 106 bp,编码701个氨基酸;山羊ACSS2基因与牛、马、人、犬、猪、小鼠和鸡的同源性分别为97.8%、92.0%、91.3%、91.3%、91.1%、88.1%和73.3%。蛋白理化性质分析结果表明,ACSS2蛋白分子质量为78.72 ku,理论等电点为6.03,属于酸性蛋白;跨膜结构和信号肽分析表明,ACSS2蛋白不含跨膜结构和信号肽;结构域分析表明,该蛋白含有1个乙酰辅酶A合成酶N端结构域。亚细胞定位分析结果表明,该蛋白主要分布在内质网（44.4%）、线粒体（33.3%）、细胞质（11.1%）和细胞核（11.1%）中。蛋白质结构预测发现ACSS2蛋白含有α-螺旋（29.10%）、延伸链（21.54%）、β-转角（9.84%）及无规则卷曲（39.52%）。实时荧光定量PCR分析结果表明,ACSS2基因在不同泌乳时期均有表达,其中在泌乳中期表达量最高,在干奶期表达量最低。本试验结果为进一步研究山羊ACSS2基因在脂质代谢过程中的功能及转录调控机制提供了参考。     

三种猪Ⅰ型干扰素基因的克隆与序列分析

根据GenBank中收录的三种猪Ⅰ型干扰素（porcine interferon,pIFN）基因序列,设计了3对特异性引物。运用RT—PCR技术从长白猪外周血淋巴细胞总RNA中扩增得到了三种猪干扰素基因,并将其克隆至pGEM-T Easy载体上。序列测定结果显示,获得的猪干扰素α1基因序列全长570bp,编码189个氨基酸,获得的猪干扰素α2基因序列全长540bp,编码179个氨基酸,获得的猪干扰素β基因序列全长563bp,编码186个氨基酸。三种猪Ⅰ型干扰素基因与已知猪Ⅰ型干扰素核苷酸序列和氨基酸序列的相似性最高可达99％～100％。     

日本结缕草ZjNAC1的克隆、转录激活活性、亚细胞定位及表达分析

采用RACE技术从日本结缕草中克隆得到ZjNAC1转录因子(GenBank No. MH428376),其开放阅读框为945bp,编码314个氨基酸。ZjNAC1编码的蛋白在N端有1个典型的NAM保守结构域,属于NAC转录因子家族。通过染色体步移的方法,获得ZjNAC1基因ATG上游1635bp序列,分析发现其包含响应脱落酸(ABA)、茉莉酸甲酯(MeJA)及逆境胁迫的作用元件。构建pGBKT7-ZjNAC1载体,转化Y2HGold酵母感受态细胞,发现ZjNAC1具有转录激活活性。农杆菌介导的35S-ZjNAC1-YFP载体注射本生烟草叶片瞬时表达结果显示,ZjNAC1定位于细胞核。实时荧光定量结果表明,ZjNAC1在日本结缕草根中表达量最高;此外,ZjNAC1的表达受10μmol/L MeJA和20%PEG4000处理的诱导,但受200μmol/L乙烯(ET)、10μmol/L ABA和300mmol/L NaCl处理的抑制。     

Understanding stakeholders’ opinions and preferences for non-native pet trade management in Florida

There is growing recognition of the link between the non-native pet trade and the introduction and establishment of invasive species due to the release and escape of non-native pets. However, it is unclear whether participants in the pet trade recognize the magnitude of this invasion risk. Successful mitigation of the pet trade invasion risk requires stakeholder support for, and participation in, regulations. We conducted 29 interviews in Florida to investigate key stakeholders’ opinions about the pet trade invasion risk and the effectiveness of potential regulations to mitigate this risk. Respondents framed the effectiveness of regulations in terms of their feasibility. Respondents also identified lack of trust and the existence of an adversarial relationship among stakeholder groups as major barriers to managing the pet trade invasion risk. Compliance with regulations may be improved if policy makers and managers utilize participatory decision-making to engage stakeholders in management of this risk.     

Cloning of Buffalo AQP9 Gene and Analysing its Expression in the Buffalo Tissues

Cloning buffalo AQP9 gene and analyzing its expression in buffalo tissues.A pair of primers was designed according to the released bovine AQP9 sequences in GenBank,which was used to clone buffalo AQP9 gene.The AQP9 gene was amplified by RT-PCR,whose nucleotide sequence and protein structure were analyzed by bioinformatics methods.The expression of AQP9 in buffalo tissues was assayed by Real-time quantitative PCR.The expression of AQP9 gene in buffalo ovary and testis tissue was detected by immunohistochemical staining method.The results showed that the cloned ORF length of buffalo AQP9 gene was 888 bp,which coded 295 amino acids.The results of multiple sequence comparison showed that the nucleotide sequence of buffalo AQP9 shared 99%,90%,97% and 88% homologeous compared with that of Bos taurus,Sus scrofa,Ovis ariessis and Homo sapiens,respectively,while shared 99%,86%,97%,83% homologeous for amino acids,respectively.Phylogenetic tree analysis indicated that AQP9 gene was highly conservative in the evolutionary process.Real-time quantitative PCR results showed that AQP9 gene expressed in buffalo liver,lung,brain,skin,testis and ovary tissues with different levels,had the most abundant expression in liver,followed by in skin and testis,less observed in lung and ovary.The results of immunohistochemical staining showed that the expression of AQP9 protein varied with the development of buffalo ovarian tissue,and gradually enhanced with follicle development.In testicular tissue,AQP9 protein expressed in spermatocyte and leydig cells of developmental stage testis.These results indicated that we had successfully cloned buffalo AQP9 gene sequences.The expression and its function of AQP9 in buffalo ovaries and testes might play an important role in follicle development and spermatogenesis.     

偃麦草种质资源外部性状变异及其形态类型的初步研究

对41份偃麦草Elytrigia repens种质资源的16个外部性状变异及其分布规律进行研究,并在对16个指标性状进行相关分析及主成分分析的基础上,对34份材料进行聚类分析。结果表明,偃麦草种内形态指标变异较大,变异系数为4.88%～33.30%,其中叶层高度变异系数最大,达33.30%,其次为旗叶长(26.03%),而种子宽度、种子长度、种子厚度、颖长、叶长变异较小,均在10.0%以下;海拔高度与偃麦草生殖枝高呈显著正相关(P0.01),与颖长呈显著负相关(P0.01),而在纬度上的相关性却呈现相反趋势;经度仅与穗下第1节间长呈显著正相关(P0.01);在对外部性状相关分析及主成分分析的基础上,通过聚类分析,在欧氏距离5.0处,34份偃麦草材料可被划分为高大型、偏高大型、特殊型、低矮型4个类型,且特殊型和低矮型均具有开发为草坪草的潜能。     

基于线性谱聚类的林地图像中枯死树监测

【目的】将基于线性谱聚类超像素的方法应用在森林病虫害防治领域,可智能监测无人机森林虫害图像中的枯死树,为森林有害生物的监测工作提供技术支撑。【方法】分别以湖北省受松材线虫与辽宁省受红脂大小蠹侵害的松林无人机图像为试验数据,首先使用线性谱聚类超像素分割算法将图像划分为多个超像素;然后基于枯死树木的颜色特征,初步提取可能为枯死树的超像素区域;最后基于枯死树木与其他干扰地物具有不同的纹理特征,计算超像素的区域密度和缝隙量,利用支持向量机对初步提取的超像素进行分类,从而检测出图像中的枯死树。【结果】基于线性谱聚类超像素和支持向量机的枯死树监测方法可有效排除与枯死树木颜色相近的其他干扰地物,较准确地提取出枯死树木。使用该方法与基于植被颜色指数的阈值分割方法、基于简单线性迭代聚类超像素和随机森林的方法,对35幅受灾松林无人机图像进行试验,并选用交并比、虚警率和漏检率3个评价指标对3种方法进行定量对比分析。结果表明,基于线性谱聚类超像素的方法监测出的枯死树区域最精确,其监测结果与人工检测结果的交并比均值大于58%,且虚警率和漏检率均优于另外2种方法。【结论】基于线性谱聚类超像素的枯死树监测方法能实现松林中枯死树的快速、准确检测及定位。     

Postulation of leaf-rust resistance genes in Czech and Slovak barley cultivars and breeding lines

Leaf‐rust resistance (Rph) genes in 61 Czech and Slovak barley cultivars and 32 breeding lines from registration trials of the Czech Republic were postulated based on their reaction to 12 isolates of Puccinia hordei with different combinations of virulence genes. Five known Rph genes (Rph2, Rph3, Rph4, Rph7, and Rph12) and one unknown Rph gene were postulated to be present in this germplasm. To corroborate this result, the pedigree of the barley accessions was analysed. Gene Rph2, as well as Rph4, originated from old European cultivars. The donor of Rph3, which has been mainly used by Czech and Slovak breeders, is ‘Ribari’ (‘Baladi 16’). Rph12 originates from barley cultivars developed in the former East Germany. Rph7 in the registered cultivar ‘Heris’ originates from ‘Forrajera’. A combination of two genes was found in 10 cultivars. Nine heterogeneous cultivars were identified; they were composed of one component with an identified Rph gene and a second component without any resistance gene. No gene for leaf rust resistance was found in 17 of the accessions tested. This study demonstrates the utility of using selected pathotypes of P. hordei for postulating Rph genes in barley.     

南极磷虾粉潜在危害因子分析及风险评价进展

南极磷虾粉是目前南极磷虾船载加工的主要产品之一,富含优质蛋白质、脂质、磷脂、虾青素和矿物质等物质,具有良好的开发利用前景。目前,南极磷虾粉主要用作水产动物饲料及提取油脂等的原料。然而,研究表明南极磷虾粉也存在潜在危害因子不明确等问题,严重限制了其应用领域和范围的拓展。因此,本文分析了南极磷虾粉潜在危害因子的来源、限量标准、检测方法等,介绍了南极磷虾粉风险评估进展,旨在为南极磷虾粉危害因子的减除和深度利用提供参考依据。     

黑龙江龙口自然保护区黑熊春季生境选择

分别于2008年4～5月和2009年4～5月,在龙口自然保护区对黑熊春季生境选择进行了研究,野外工作共设置87个10 m×10 m样方,并调查12个生态因子,其中利用样方41个,对照样方46个。通过Vanderploeg ＆Scavia选择指数得到,黑熊喜爱选择处于高海拔（〉450 m）、上坡位有石砬子且草本盖度适中（10%～40%）的生境,倾向选择乔木密度高（〉10株/100 m2）、郁闭度大（〉40%）、灌木密度较低（〈10%）、隐蔽级低（〈30%）、倒木密度大（〉1根/100 m2）、陡坡（〉30°）且向阳的针阔混交林。主成分分析与资源选择函数分析共同得到海拔、坡度、坡位、植被类型对黑熊生境选择影响较大,坡向和倒木密度影响较小的结果。综合分析,将Vanderploeg ＆ Scavia选择指数、主成分分析与资源选择函数相结合可以更好地反映黑熊生境选择情况。