Viral RNA extraction using an automatic nucleic acid extractor with magnetic particles and genetic characterization of bovine viral diarrhea virus in Tokachi Province,Japan, in 2016–2017

In this study, the viral genome extraction performance of automatic nucleic acid extractors and manual nucleic acid extraction kits was compared. We showed that compared with manual kits, the automatic extractors showed superior genome extraction performance using bovine viral diarrhea virus (BVDV) genome-positive cattle sera and bovine coronavirus/infectious bovine rhinotracheitis virus-spiked cattle nasal swabs. In addition, the subgenotyping of BVDV strains detected in Tokachi Province in Japan during 2016–2017 was performed. Results showed that most of these BVDV strains belonged to subgenotype 1b, while few strains belonged to subgenotypes 1a and 2a. This study showed the high applicability of automatic nucleic acid extractors in extracting multiple viral genomes and the dominant subgenotype of BVDV in Tokachi.     

A morphological analysis of the skull size and shape of Kerivoulinae (Chiroptera: Vespertilionidae) from Vietnam

Southeast Asia is a region of high biodiversity, containing species of plants and animals that are yet to be discovered. In this region, bats of the subfamily Kerivoulinae are widespread and diverse with six species recorded in Vietnam. However, the taxonomy of the Kerivoulinae in Asia is complicated. In our study, we used diagnostic characters and multivariate analysis to determine morphological differences between the genera Kerivoula and Phoniscus. We showed that the two genera are distinguishable by the size of second upper incisors, the shape of skull, nasal sinus, canines, second upper and lower premolars. In addition, the two genera can be osteometrically separated by measurements of the braincase height, interorbital width and shape of anterior palatal emargination. Our data clearly revealed the morphological variations in the skull shape of Kerivoula hardwickii in Vietnam. This suggests a possible separation into three morphotypes, representing cryptic species supported by statistical differences with wide variation in skull shape, size and teeth. These results demonstrated Kerivoula hardwickii can be separated three subspecies, and the result will serve as the basis for the future assessment and classification of this group in Southeast Asia.     

年龄、温度、膘情对伊犁马卵泡发育的影响

【目的】 研究新疆昭苏地区伊犁马发情周期内卵泡发育变化规律,分析年龄、温度、膘情对伊犁马母马卵泡发育的影响,为掌握伊犁马母马卵泡发育规律提供理论依据。【方法】 选取伊犁哈萨克自治州昭苏县康苏马业配种站内,同一草场放牧情况下84匹母马作为研究对象监测发情期内卵泡发育,分析对比不同年龄、温度、膘情对伊犁马卵泡发育时长的差异性。【结果】 年龄Ⅱ组TF3-F5显著高于年龄Ⅲ组(P＜0.05)。年龄Ⅱ组TF4-5显著高于年龄Ⅲ组(P＜0.05)。平均高温对TF1-F2具有显著影响(P＜0.05),平均高温Ⅰ组TF1-F2显著高于平均高温Ⅱ组(P＜0.05)。平均低温对TF1-F2具有极显著影响(P＜0.01),平均低温Ⅰ组TF1-F2极显著高于平均低温Ⅱ组(P＜0.01)。积温对TF1-F2具有显著影响(P＜0.05),积温Ⅱ组TF1-F2极显著高于积温Ⅰ组(P＜0.01)。膘情对TF1-F2、TF3-F4、TF1-F5、TF2-F5、TF3-F5均具有极显著影响(P＜0.01),对TF2-F3具有显著影响(P＜0.05)。膘情Ⅰ组TF1-F2极显著高于膘情Ⅲ组(P＜0.01)。膘情Ⅰ组TF2-F3显著高于膘情Ⅲ组(P＜0.05)。膘情Ⅲ组TF3-F4极显著低于膘情Ⅰ组(P＜0.01),膘情Ⅲ组TF3-F4显著低于膘情Ⅰ组、膘情Ⅱ组(P＜0.05)。膘情Ⅰ组TF1-F5极显著高于膘情Ⅱ组、膘情Ⅲ组(P＜0.01),膘情Ⅱ组TF1-F5显著高于膘情Ⅲ组(P＜0.05)。膘情Ⅰ组TF2-F5极显著高于第Ⅲ组(P＜0.01),膘情Ⅰ组TF2-F5显著高于膘情Ⅱ组、膘情Ⅲ组(P＜0.05)。【结论】 年龄、温度、膘情均一定程度的影响伊犁马母马的卵泡发育,其中膘情对卵泡发育有极显著影响,体况较差可抑制母马卵泡发育。     

甜瓜属野生种Cucumis hystrix Chakr.胞质葡萄糖-6-磷酸脱氢酶基因的克隆与启动子结构分析

根据植物葡萄糖-6-磷酸脱氢酶(G6PDH,EC1.1.1.49)氨基酸保守区域,设计简并引物,采用RT-PCR技术克隆得到甜瓜属野生种Cucumis hystrix Chakr.(2n=24)G6PDH基因cDNA片段;随后基于该序列设计特异引物,PCR方法筛选野生种BAC文库,获得2个BAC阳性单克隆。测序后获得了G6PDH基因全序列及其上游启动子序列,GenBank登录号:JQ771576。序列分析显示:G6PDH基因全长约6.5 kb,由15个外显子和14个内含子组成;内含子序列均符合5’-gt-ag-3’结构。外显子拼接后获得了G6PDH基因的开放阅读框(ORF)序列,序列全长1 551 bp,编码516个氨基酸,与黄瓜基因组网站公布的G6PDH基因核苷酸序列和氨基酸序列同源性分别为98.71%和99.03%。野生种G6PDH氨基酸序列N端缺少转运肽序列,确定为野生种胞质G6PDH。氨基酸序列与烟草、马铃薯、荷兰芹、猕猴桃、拟南芥、大豆、小麦、玉米、葡萄、杨树等植物胞质G6PDH同源性高达77%以上。系统发育树分析发现,甜瓜属胞质G6PDH与茄科植物最先聚类,植物胞质G6PDH在分子进化水平上与物种进化相符。启动子结构分析表明:序列含有启动子基本元件TATA-box、CAAT-box,还含有丰富的光响应元件,与环境胁迫响应相关的不同顺式作用元件,促进高水平转录的5UTR Py-rich stretch元件和提高表达的CAT-box元件等。     

杂交粳稻亲本穗长和枝梗数性状优异配合力的标记基因型筛选

提高杂交粳稻竞争优势的关键是选育具有高配合力的大穗型杂交粳稻恢复系.为使配合力改良更有效,选用115对SSR引物扩增6个粳稻BT型不育系和12个恢复系的标记基因型,并按NC Ⅱ(Noah Carolina Ⅱ)遗传交配设计配制72个F1,组合,分析18个亲本的一次枝梗数、二次枝梗数和穗长3个穗部性状的配合力,结合亲本SSR分子标记数据和性状配合力数据筛选3个穗部性状优异配合力的标记基因型.结果发现:23个标记基因型与亲本3个穗部性状配合力显著相关.其中1个标记基因型与亲本3个性状配合力都相关;7个标记基因型与亲本2个性状配合力都相关;15个标记基因型与亲本单个性状配合力相关.标记基因型RM8254-120/180对3个性状的配合力效应都是正的,可使F1的每穗一次枝梗数、二次枝梗数和穗长3个性状值分别增加21.63%、21.12%和16.12%.与亲本2个性状配合力相关的7个标记基因型中有6个标记基因型对亲本的配合力效应是正值,1个标记基因型对亲本的配合力效应是负值.     

Re-evaluation of total sulphur amino acid requirement and determination of replacement value of cystine for methionine in semi-purified diets of juvenile Nile tilapia, Oreochromis niloticus

Two feeding experiments were conducted to re-evaluate the total sulphur amino acid (TSAA; methionine and cystine) requirement and determine the replacement value of cystine for methionine of juvenile Nile tilapia ( Oreochromis niloticus ). Semi-purified diets used in both experiments contained 3510 kcal gross energy and 280 g of protein per kilogram diet from casein, gelatin and crystalline amino acids. The basal diet of the first experiment contained 3.1 g methionine and 0.4 g cystine per kilogram. l -methionine was added to the seven remaining diets at 1.0 g kg⁻¹ increment to produce methionine levels ranging from 3.1 to 10.1 g kg⁻¹ diet. Each diet was fed to four replicate groups of juvenile Nile tilapia (1.28 g mean weight) in a recirculation system for 8 weeks. Broken-line regression analysis of weight gain data indicated that the TSAA requirement of juvenile Nile tilapia was 8.5 g kg⁻¹ of the diet or 30.4 g kg⁻¹ of dietary protein. In the second experiment, TSAA level was set at 95% of the requirement value determined in the first experiment. Seven diets were made with different ratios of l -methionine and l -cystine (20 : 80, 30 : 70, 40 : 60, 50 : 50, 60 : 40, 70 : 30 and 80 : 20, based on an equimolar sulphur basis). Each diet was also fed to four replicate groups of juvenile Nile tilapia (4.14 g mean weight) in a recirculation system for 8 weeks. Regression analysis of weight gain data using broken-line model indicated that cystine (on a molar sulphur basis) could replace up to 49% of methionine requirement in semi-purified diets for juvenile Nile tilapia.