Detection of citrus psorosis-ringspot virus using RT-PCR and DAS-ELISA

Psorosis, sometimes also associated with ringspot symptoms, is a widespread and damaging disease of citrus in many parts of the world including South America and the Mediterranean basin. We describe the application of RT-PCR and DAS-ELISA diagnostics to an isolate of citrus ringspot virus (CtRSV-4) and other virus isolates associated with this disease. Fragments of cDNA from bottom-component RNA of CtRSV-4 were cloned and sequenced, and PCR primers were designed, 5'ACAATAAGCAAGACAAC upstream, and 5'CCATGTCACTTCTATTC downstream. RT-PCR experiments using these primers allowed detection of CtRSV-4 in infected citrus leaves down to a tissue dilution of 1/12 800 representing 2 μg of tissue, and less sensitive detection of the related citrus psorosis-associated virus (CPsAV90-1-1) and four other psorosis isolates from Argentina and the USA. In addition, CtRSV-4 particles were partially purified from local lesions in Chenopodium quinoa, and the preparations used to raise a rabbit antiserum. The antiserum was absorbed with extracts of healthy C. quinoa leaves, and a DAS-ELISA kit was prepared and tested for detection of CtRSV-4, CPsAV90–1-1, and other psorosis isolates from Argentina, the USA, Italy and Spain. The ELISA detected CtRSV-4 down to a tissue dilution of 1/1600, and most other psorosis isolates down to dilutions of 1/200–1/800. Three of a total of 20 heterologous isolates were consistently negative. Comparison of the PCR and ELISA results suggests that both methods can be used for detection of a range of psorosis isolates, but that variation of the viruses in the field might cause problems for any one diagnostic test.     

镉在黄瓜幼苗中的化学形态及亚细胞分布

为探究黄瓜幼苗对镉的吸收累积特性,将黄瓜幼苗置于6种镉浓度梯度(0、10、50、100、200、300μmol·L~(-1))营养液中培养17 d,测定黄瓜幼苗地上部和根部的镉含量以及镉在叶片的亚细胞分布和其化学形态分布。结果表明:随镉浓度升高,转移系数逐渐降低,黄瓜由根向地上部对镉的转运能力减弱。时间或浓度处理均会显著增加黄瓜叶片各亚细胞组分镉含量,镉的富集比例趋势表现为可溶性部分细胞壁细胞器,其中可溶性部分镉含量占44.88%～84.94%,远大于细胞器(3.10%～17.05%)与细胞壁(11.20%～39.66%)。相同时间下,随镉处理浓度的增加,镉在细胞壁组分占比先升高后降低,可溶性部分镉的占比先降低后升高。同时随着处理时间延长,细胞壁组分镉占比降低,可溶性部分镉占比升高。黄瓜叶片中NaCl提取态镉占比最高(33.69%～54.23%),但随着镉浓度提高,乙醇提取态、H2O提取态镉占比升高。当镉浓度达到300μmol·L~(-1)时,黄瓜叶片中的镉形态以乙醇提取态、H2O提取态和NaCl提取态占优势。总的来看,在器官水平上,黄瓜通过限制镉由根部向地上部传递的方式,降低镉对黄瓜地上部的毒害;亚细胞水平上,细胞壁对镉的吸持和液泡区室化是黄瓜对镉胁迫下的重要解毒方式,其中以液泡的区室化占主导位置;在化学形态分布上,镉的化学形态转化也是黄瓜应对镉胁迫的重要耐性策略。     

贵南县高寒草地荒漠化及其控制技术与措施

通过简述贵南县高寒干旱区草地类型、分布区域及荒漠化特征 ,从气候变化、鼠害、盲目开垦及超载过牧等自然、人为因素方面 ,分析总结了草地荒漠化引发机理 ,提出了防止草地荒漠化的对策及治理技术。     

低磷胁迫对小麦镉吸收的影响

为探究低磷胁迫下小麦根系生理和形态的变化及其对镉的吸收能力的影响,以Ca₃（PO₄）₂作为磷源,通过砂培试验研究了在低磷胁迫下小麦根系形态和分泌物变化特征,以及这些变化对难溶态镉（CdCO₃）的活化与吸收的影响。结果表明,在低磷处理中,小麦地下部生物量显著低于对照,降幅为27.3%（P<0.05）,根长度、根表面积和根体积均显著变小（P<0.05）。此外,低磷胁迫下小麦地上部和地下部磷含量均显著降低,降幅分别为35.4%和23.1%（P<0.05）,但是显著促进了难溶态磷的溶解（P<0.05）。同时还发现低磷胁迫显著提高了小麦植株Cd含量和Cd的总活化量,增幅分别为190.8%和82.8%（P<0.05）。低磷胁迫下培养基pH降低了0.3,同时根中草酸根和苹果酸根含量显著升高,增幅分别为1 588.1%和37.7%（P<0.05）,由此可见,低磷胁迫下小麦通过分泌质子和羧酸根促进了磷的活化,并促进了Cd的活化,进而导致小麦Cd吸收的增加。这些结果阐明了低磷胁迫影响小麦Cd吸收的机制。     

Standing Laparoscopic Peritoneal Flap Hernioplasty of the Vaginal Rings does not Modify the Sperm Production and Motility Characteristics in Intact Male Horses

Laparoscopic hernioplasty techniques have been developed in the recent years to avoid the recurrence of inguinal hernias and to spare the testicles for breeding purposes in stallions. However, there have been no previous comprehensive and systematic studies of the reproductive outcomes and prognoses for stallions after inguinal hernioplasty. Therefore, the objective of this study was to assess the possible effects of one of these techniques (standing laparoscopic peritoneal flap hernioplasty) on the sperm production and motility characteristics of six healthy stallions that received this procedure based on 1‐year follow‐ups. There were no significant differences in the measured sperm variables (assessments based on the DSO, MOT, PMOT, VSL, VCL and VAP) during 1‐year follow‐ups.     

西藏不同青稞品种的品质差异分析

为了解西藏地区不同青稞品种的品质差异,采集了西藏青稞主产区的不同青稞品种,并对青稞中的水分、淀粉、粗蛋白、粗纤维、矿质元素以及有效成分总黄酮、花青素、β-葡聚糖等14项指标进行了检测。结果表明:有色青稞中淀粉和微量元素Mn、Cu、Zn的含量在年度间存在着显著的差异,且有色青稞微量元素含量普遍偏低;青稞总黄酮含量范围在0.14%~0.42%,均值为0.23%;β-葡聚糖含量范围为2.65%~6.52%,均值为4.59%;有色青稞花青素含量变化较大,范围在0.022 7~0.649 7 mg/g,均值为0.133 2 mg/g。总黄酮含量、β-葡聚糖含量年际间差异不大,花青素含量年际间差异巨大。总黄酮含量有色青稞显著高于普通青稞,而β-葡聚糖含量二者无显著差异。     

液相色谱质谱联用分离、鉴定芥蓝中脱硫芥子油苷

    从芥蓝菜薹中提取芥子油苷,经阴离子交换柱DEAE-Sephadex A-25纯化和硫酸酯酶脱硫,得到脱硫芥子油苷提取液.比较了两种高效液相色谱的流动相体系(甲醇与水体系和乙腈与水体系)及两种长度(250 mm×4.6 mm和150 mm×4.6 mm)的C18色谱柱(Prontosil ODS2,粒径5μm)对芥蓝脱硫芥子油苷的分离效果,确定分离条件为乙腈与水流动相体系和250 mm×4.6 mm C18柱.采用质谱联用及紫外扫描,在芥蓝菜薹中检测出7种脂肪族芥子油苷和4种吲哚族芥子油苷,其中4-甲硫基丁基芥子油苷为第一次在芥蓝中检测到.芥蓝中主要以脂肪族芥子油苷为主,占总芥子油苷含量的91.6%,而吲哚组芥子油苷的含量仅占8.4%.其中,3-丁烯基芥子油苷和4-甲基亚硫酰丁基芥子油苷为主要芥子油苷组分,分别占总含量的47.9%和35.5%,而含量最低的为4-羟基-3-吲哚甲基芥子油苷,只占总芥子油苷含量的0.2%.