Secretoglobin and Transferrin Expression in Bronchoalveolar Lavage Fluid of Horses with Chronic Respiratory Disease

Background

Lower expression of secretoglobin and transferrin has been found in the bronchoalveolar lavage fluid (BALF) of a small number of horses with experimentally induced signs of recurrent airway obstruction (RAO) compared to healthy controls.

Hypothesis/Objectives

Secretoglobin and transferrin BALF expression will be similarly decreased in horses with naturally occurring clinical signs of RAO and in horses with experimentally induced clinical signs of RAO as compared to healthy controls and intermediate in horses with inflammatory airway disease (IAD).

Animals

Recurrent airway obstruction‐affected and control horses were subjected to an experimental hay exposure trial to induce signs of RAO. Client‐owned horses with a presumptive diagnosis of RAO and controls from the same stable environments were recruited.

Methods

Pulmonary function and BALF were evaluated from control and RAO‐affected research horses during an experimental hay exposure trial (n = 5 in each group) and from client‐owned horses (RAO‐affected horses, n = 17; IAD‐affected horses, n = 19; healthy controls, n = 5). The concentrations of secretoglobin and transferrin in BALF were assessed using Western blots.

Results

Naturally occurring and experimentally induced RAO horses had similar decreases in BALF transferrin expression, but secretoglobin expression was most decreased in naturally occurring RAO. Secretoglobin and transferrin expression were both lower in BALF of RAO‐affected horses than in IAD‐affected and control horses.

Conclusions and Clinical Importance

Secretoglobin and transferrin expression is decreased in BALF of RAO‐affected horses after both experimental and natural exposure. Secretoglobin and transferrin likely play clinically relevant roles in the pathophysiology of RAO, and may thus be used as biomarkers of the disease.     

卷丹百合远缘杂交及杂种后代的鉴定

为培育抗性强、观赏性优良的卷丹百合新种质,采用液体培养基悬滴法测定3个观赏百合品种‘Black stone’‘Tresor’‘Sorbonne’ 的花粉生活力;用蕾期授粉、正常授粉、延迟授粉、切割柱头授粉方式分别与母本卷丹百合进行杂交,对获得的杂交后代进行胚珠培养及增殖扩繁,采用SSR分子标记的方法鉴定杂种的真实性。结果表明：‘Sorbonne’的花粉萌发率总体高于‘Black stone’和‘Tresor’;蕾期授粉的结实率最高,为14.44%;卷丹בBlack stone’ 组合获得蒴果数量最多,共得到了17个蒴果;对杂交蒴果进行胚珠培养,得到9株F1代杂种苗;筛选杂种苗增殖扩繁的较适培养基为MS+30 g/L蔗糖+0. 1 mg/L NAA+1 mg/L 6–BA+8 g/L琼脂;鳞茎膨大生根较适培养基为MS+80 g/L蔗糖+0.1 mg/L NAA+ 0. 05 mg/L 6–BA+0.75 g/L活性炭+6.5 g/L琼脂,最适培养环境为黑暗培养;通过4对引物鉴定证实卷丹בTresor’的2个杂种株系为真实杂种。     

Sexual compatibility of transgenic soybean and different wild soybean populations

The introduction of genetically modified (GM) soybean into farming systems raises great concern that transgenes from GM soybean may flow to endemic wild soybean via pollen. This may increase the weediness of transgenic soybean by increasing the fitness of hybrids under certain conditions and threaten the genetic diversity of wild soybean populations. Although pollen-mediated gene flow between GM crops and wild relatives is dependent on many factors, the sexual compatibility (SC) determined by their genetic backgrounds is the conclusive factor. The considerable genetic variation among wild soybean populations may cause compatibility differences between different wild and cultivated soybeans. Thus, an evaluation of the SC between transgenic soybean and different wild soybeans is essential for assessing the environmental consequences of cultivated soybean–wild soybean transgene flow. The podding and seed sets were assessed after artificial hybridization using transgenic glyphosate-resistant soybean as the paternal parent and 18 wild soybean populations as the maternal parents. Then, the average number of filled seeds produced in 200 flowers (AFS) was calculated for each wild soybean under natural self-pollination as well as under artificial crossing with transgenic soybean. Finally, the index of cross-SC was calculated (ICSC) as the ratio of the AFS of wild soybean artificially crossed with transgenic soybean and the AFS of naturally self-pollinated wild soybean. The results demonstrated that after self-pollination and crossing with transgenic soybean, the average podding rates of 18 wild soybean populations ranged within 96.50–99.50% and 4.92–18.03%, and the average filled seed numbers per pod varied from 1.70 to 2.69 and 0.20 to 0.48, respectively. The results showed that approximately 89% of wild soybeans displayed either medium or higher than medium SC with transgenic soybean (ICSC>1.0%). This implied the high possibility of gene flow via pollen from transgenic soybean to wild soybean.     

白菜基因组导入芥菜的远缘杂交及后代分离偏向性

【目的】研究远缘杂交将结球白菜的A基因组导入包心芥菜,利用高分辨率熔解曲线(high-resolution melting, HRM)结合InDel标记辅助选择技术,筛选出保留白菜基因较多的回交后代,改良包心芥的结球性,丰富包心芥菜的种质资源。【方法】利用幼胚培养技术辅助6份包心芥菜与2份结球白菜进行远缘杂交和回交,选取20个InDel标记追踪白菜A基因组全部染色体,从192的BC₁单株中筛选出白菜A基因组标记占比较高的单株。田间调查筛选结球性近似结球白菜的单株。【结果】远缘杂交结合幼胚培养获得了9个种间杂种组合和14个BC₁群体。筛选并获得203个在全基因组分布均匀的可区分白菜与芥菜的HRM-InDel标记;利用20个InDel标记追踪192株BC₁单株的白菜基因,分离群体偏向传递白菜基因组,有54棵单株标记含量超过90%。【结论】幼胚培养技术可提高白菜与芥菜远缘杂交后回交的结实率,并可加快育种进程;筛选出了白菜基因组占比较高的组合和单株;分离群体明显偏向于传递白菜基因。     

基于逆向工程的搅浆刀作业性能界定与关键设计参数研究

水田搅浆是基于稻作制度的独立机械化作业环节,搅浆刀是进行搅浆农艺处理的专用刀具,但目前缺乏针对搅浆的农艺界定及搅浆刀专用设计参数研究。本文利用逆向工程技术获取搅浆刀的关键几何参数,并对搅浆作业和搅浆刀的设计方法进行定义。通过与常规旋耕刀(R245)进行分析对比提出搅浆刀的专用设计参数,进而通过田间试验进一步探讨搅浆刀的设计方法。结果表明,搅浆刀的楔面入土效应是保障其水田搅浆性能的关键,该效应所对应的搅浆刀设计参数被定义为侧切刃楔角,侧切刃楔角与侧切刃刀面宽度2个关键设计参数决定了搅浆刀入土过程的侧向搅浆作业强度。与R245旋耕刀相比,侧切刃楔角是搅浆刀设计的显著特点,搅浆刀的侧切刃静态楔角高于动态楔角,而R245的侧切刀面是一平直刀面(楔角为0°),在正切刃区域,搅浆刀与旋耕刀的楔角变化基本一致。基于田间土壤条件的原位台架试验进一步证明了搅浆刀专用设计参数对保障起浆、水田埋草等性能的优越性。     

模拟降雨下重庆喀斯特槽谷区径流氮磷元素流失特征

为揭示降雨强度和岩层倾向对喀斯特槽谷区耕层土壤氮磷流失的影响,利用人工降雨装置和可变坡钢槽进行模拟试验,设计坡面坡度为25°,岩石裸露率为15%,岩层倾角为30°,地下孔裂隙度为3%,降雨强度分别为30、60和90mm/h,模拟降雨持续时间为1h。结果表明:1)产流量随着降雨强度的增大而增大,地表产流量表现为裸坡顺层坡逆层坡;地下产流量表现为逆层坡顺层坡裸坡。2)在30mm/h降雨强度条件下径流全氮浓度最高;在60mm/h降雨强度条件下铵态氮浓度和硝态氮浓度最高;磷浓度随着降雨强度的增大而降低;逆层坡的径流氮磷浓度最高。3)氮磷流失量随降雨强度增大而增大,且地下流失量高于地上流失量;不同岩层倾向条件下,氮磷流失量地上表现为裸坡顺层坡逆层坡,地下表现为逆层坡顺层坡裸坡;氮在逆层坡的流失量最多,磷在顺层坡的流失量最多。本研究可为在喀斯特槽谷区布设保水保土措施以及改善土壤质量上针对不同岩层倾向坡方案提供依据。     

关于贝类遗传改良几个问题的讨论

贝类大多为雌雄异体，少数种类是雌雄同体，在雌雄异体的种类也有部分个体表现为雌雄同体，而且雌雄异体型贝类还常有性转变现象。养殖贝类大多行体外受精并具有配子量大、子裔数量多等特点。野生型贝类表型方差大，遗传变异水平高。可见，贝类是一群比较独特的动物，其生殖方面的某些特性与作物的共同点甚至超过与高等动物的共同点。因此仅依靠脊椎动物或植物的育种方法，难以解决好贝类的种质改良问题。     

Production of polyclonal antiserum against recombinant VP28 protein and its application for the detection of white spot syndrome virus in crustaceans

The VP28 gene of white spot syndrome virus (WSSV) was cloned into pRSET B expression vector. The VP28 protein was expressed as a protein with a 6-histidine taq in Escherichia coli GJ1158 with NaCl induction. Antiserum was raised against this recombinant-VP28 protein in rabbits and it recognized VP28 protein in naturally and experimentally WSSV-infected shrimp, marine crabs, freshwater prawns and freshwater crabs. The antiserum did not recognize any of the other known WSSV structural proteins. Various organs such as eyestalks, head muscle, gill tissue, heart tissue, haemolymph, tail tissue and appendages were found to be good materials for detection of WSSV using the antiserum and detection of WSSV was successful in experimentally infected Penaeus monodon and P. indicus at 12 and 24 h post-infection (p.i.), respectively. The antiserum was capable of detecting WSSV in 5 ng of total haemolymph protein from WSSV-infected shrimp.     

杂色鲍与盘鲍种间杂交受精率的影响因素

研究卵子排放后的时间、精子排放后的时间、授精精子浓度、胰蛋白酶、钙离子、pH和水温等因子对杂色鲍(Haiotisdiversicolor,以下简称S)与盘鲍(H.discus discu,以下简称J)种间杂交受精率的影响。结果表明,卵子排放后的时间和授精精子浓度是影响杂交受精率两个最重要的因素。与自繁组合相比,正反杂交受精率均较低且下降速度快得多。水温21.5℃时,两亲本自繁对照组中,卵子排放1 h对受精率均无显著影响。而SJ杂交,卵子排放时间2 min时,受精率为55.9%±8.1%,4.7 min时受精率已下降了50%;JS杂交,卵子排放时间为4 min时,受精率为31.5%±9.7%,10 min时受精率下降至4 min时的29.1%。相比之下,精子排放时间对杂交受精率的影响小得多,3 h内对正反交杂交受精率没有显著影响。杂交适宜精子浓度约为母本自繁的100倍,其中SJ杂交适宜授精精子浓度为6.0×106~2.4×107mL-1,JS杂交适宜授精精子浓度为3.8×107~7.5×107mL-1。水温对于杂交受精率也有影响,短时间内将水温控制在26~29℃可以得到较高受精率。其他实验因素,如添加胰蛋白酶、Ca2+或改变授精体系pH对杂交受精率均无正向作用。     

抑制消减杂交（SSH）及其在鱼类基因克隆中的应用

和其它脊椎动物一样,在鱼类的生命进程中,某种特定类型的细胞中只有比例约为10%～15%的基因表达.由于这些基因的特异性决定了从受精卵到机体死亡的整个生命过程,所以这些基因的时空表达是受到严格调控的.在不同发育阶段,不同生理状态和不同类型的细胞中,表达的基因各不相同.因此,比较两种不同类型细胞中基因表达的差异,可以探究表型的遗传原因,了解生命过程的基本信息,从而找到与发育、疾病等相关的基因.