甜菜SSR反应体系优化及重要农艺性状分子标记

[目的]为选育优质甜菜新品种,利用SSR法筛选与高糖、高产、耐盐相关的分子标注.[方法]研究对甜菜SSR-PCR反应体系进行优化,并利用SSR分子标记方法和分离群体分组分析法(Bulked SegregateAnalysis,BSA)对具有高糖/低糖、低产/高产、耐盐/不耐盐三种重要农艺性状的甜菜亲本和F2代植株进行分析.[结果]研究建立了适宜甜菜的SSR-PCR反应体系为:20μL反应体系中含l×Buffer、2.0 mmol/LMg2+、1.5 uTaq DNA聚合酶、0.20 mmol/L dNTP、1.5 μmol/L引物、60 ng DNA模板.依据优化体系,对不同农艺性状的甜菜亲本与F2代进行SSR-PCR扩增分析,高糖性状获得了200和100 bp两条与高糖性状连锁的标记,250和230 bp两条与高产性状连锁的标记,550、250和100 bp三条与耐盐性状紧密连锁的分子标记.[结论]研究获得的7条特异条带是与甜菜重要农艺性状连锁的分子标记,将为甜菜的育种工作提供重要的理论基础.     

簇毛麦抗白粉病基因的RAPD及RFLP标记

 分析了来自前苏联簇毛麦及其抗病衍生系的抗白粉病基因对不同生理小种的抗性反应。用120个随机引物对6D/6V代换系Pm930640进行RAPD分析，检测到5个引物OPAN03、OPAI01、OPAL03、OPAD07和OPAG15，分别在大约1 700、700、750、480和580 bp处有区别于小麦亲本的多态性条带。对Chancellor×Pm930640 F2群体进行OPAN03、OPAI01和OPAL03等3个RAPD标记与抗白粉病基因的连锁分析，表明这些标记同簇毛麦的抗白粉病基因是连锁的。对大部分分别含有Pm1－Pm20的已知抗病基因、含有簇毛麦抗病基因及其相关亲本的29个小麦品系进行RAPD标记分析。结果表明，这些标记不仅可以鉴定簇毛麦的抗病基因，而且可以判断其遗传背景。OPAL03750仅出现在含有前苏联簇毛麦6VS染色体的抗病材料中，可作为区别于Pm21的分子标记。RFLP标记的结果也表明两个不同簇毛麦的6VS染色体有明显的多态性。     

青海不同产区大黄的X-射线衍射图谱研究（摘要）

[目的]为区分不同产地大黄药材的质量及鉴别提供依据。[方法]将供试材料青海玉树掌叶大黄（样品1）、青海海南唐古特大黄（样品2）与青海果洛唐古特大黄（样品3）分别粉碎成粉末,过100目筛,即可供X射线衍射试验用。设定管压30 kV,管流20 mA,2θ扫描范围3°～90°,扫描速度0.06°/s,每种样时间0.5 s,获得3种不同产区大黄的X-射线衍射图谱。试验数据以晶面间距d与特征峰相对强度L/Io表示,记为d/（I/Io）。将试验数据导入中药指纹图谱相似度计算软件,经选峰,设定匹配模板,将谱峰自动匹配,然后设定标准模板,进行谱峰差异性评价和整体相似性评价。[结果]样品1中所含的化学成分与样品2相同,但峰的强度（I/Io）不同,表明相同成分在两种样品中的含量不相等。同样把样品3的图谱与其他两个样品进行比较,也会得到相同的衍射峰值。表明不同产区的大黄中各化学成分的含量有差异,但其衍射图谱及衍射峰值具有一定的指纹特征。[结论]X-射线衍射法是鉴别不同产区的大黄及其他中药材的一种快捷有效的方法。     

绵羊LDHβ基因的生物信息学分析及其在不同剩余采食量绵羊肝脏和肌肉组织中的表达

【目的】为鉴定绵羊乳酸脱氢酶β(lactate dehydrogenase B,LDHβ)基因的分子特征,研究其在不同剩余采食量绵羊肝脏和肌肉组织中的表达差异.【方法】测定了137只‘湖羊’公羔的剩余采食量(residual feed intake,RFI),按RFI进行排序,分别筛选出RFI最高(high-residual feed intake,H-RFI)和RFI最低(low-residual feed intake,L-RFI)的羊各15只,屠宰后,采集肝脏和肌肉组织,利用Q-PCR技术检测绵羊LDHβ基因分别在H-RFI和L-RFI羊肝脏和肌肉中表达量,并利用生物信息学软件构建了该基因的系统进化树和预测其结构与功能.【结果】绵羊LDHβ基因开放阅读框(open reading frame,ORF)为1 005bp,编码334个氨基酸,其蛋白质分子质量为36 479.43U,理论等电点为6.40.绵羊LDHβ基因与山羊的亲缘关系较近,其次为牛,序列同源性高.功能结构域预测结果显示,该基因编码产物在内质网中参与辅酶因子生物合成的可能性最高.Q-PCR结果显示,绵羊LDHβ-mRNA基因在L-RFI羊肝脏和肌肉中表达量均显著低于H-RFI羊(P0.01).【结论】绵羊LDHβ基因作为能量代谢的关键酶参与绵羊饲料效率的调控.     

The(13)C-octanoic acid breath test for detection of effects of meal composition on the rate of solid-phase gastric emptying in ponies

The aim of this study was to apply the(13)C-octanoic acid breath test for detection of alterations in the rate of solid-phase gastric emptying, induced by changes in test meal composition, in ponies. After a 14 hour fast the ponies (n = 4) ingested a test meal with 0, 35 or 70 ml soya oil, and labelled with 250 mg(13)C-octanoic acid. Each pony was given each of the three test meals on three separate occasions, in a randomised order. Exhaled breath samples were collected for 12 hours after ingestion of the test meal. Breath samples were analysed by continuous flow isotope ratio mass spectrometry. Three indices of breath(13)C-enrichment were computed, half-dose recovery time (t 1/2), gastric emptying coefficient (GEC) and time to peak breath(13)C-excretion t(max). The(13)C-octanoic acid breath test was a reliable means of assessing the significantly decreased rate of gastric emptying in the pony, associated with addition of soya oil to the test meal.     

Arthroscopic laser extirpation of metacarpophalangeal synovial pad proliferation in eleven horses

A new surgical technique for treatment of chronic metacarpophalangeal synovial pad proliferation in the horse and the findings and long-term follow-up from 11 clinical cases are described. The medical records of all equine lameness cases attributed to metacarpophalangeal synovial pad proliferation admitted to the College of Veterinary Medicine at Cornell University (1991-1996) were reviewed and all those treated surgically by laser extirpation were included in this study. Retrieved data included subject details, preoperative lameness, ultrasonography, radiography and synovial fluid evaluations and lesion histopathology. Lesions were ablated using a CO2 or a Nd:YAG laser intra-articularly with arthroscopic guidance. Long-term follow-up was provided by telephone conversation with owners or trainers. All horses had fetlock joint effusion and were lame at presentation. Mean synovial pad thickness measured ultrasonographically was 9.0 mm (range 6-15 mm). Seven horses (64%) had radiographic evidence of remodelling of the dorsal cortex of distal McIII and 3 horses (27%) had concurrent dorsal proximal P1 fractures. No postoperative complications were noted. All 11 horses returned to training within 90 days of surgery without recurrence of the lesion(s). Laser extirpation of metacarpophalangeal synovial pad proliferation using arthroscopic guidance provided a rapid, safe and efficient method for surgical removal of such lesions without complications or recurrence. This surgical technique provides a suitable alternative to more conventional treatments for chronic metacarpophalangeal synovial pad proliferation in horses, particularly for removal of very large, fibrotic masses.     

Reinvestigating adjuvants for the wild oat herbicide,flamprop-M-isopropyl. II: Field performance

A field trials programme was conducted in which the performance of a new emulsifiable concentrate formulation (ECI) of flamprop-M-isopropyl containing the adjuvant, ‘Dobanol’ 25-7, in a ratio of 2:1 (by weight) with the AI, was compared with the current commercial formulation of ‘Commando’, in combination with its recommended adjuvant, ‘Swirl’, for the control of wild oat (Avena fatua L.) in wheat (Triticum aestivum L.) and barley (Hordeum vulgare L.). A further treatment, in which the ‘Dobanol’ 25-7: AI ratio was increased to 4:1 by the spray tank addition of the former, was also included. The mean results from six trials (five wheat, one barley) showed that the addition of ‘Swirl’ to ‘Commando’ was beneficial, increasing wild oat floret control from a mean value of 80% to 92% at current recommended rates (flamprop-M-isopropyl, 600 g ha^?1; ‘Swirl’, 2.5 litre ha^?1). However, combinations of flamprop-M-isopropyl and ‘Dobanol’ 25-7 gave superior levels of control even at lower AI application rates. For example, a mean level of 96% control of Avena spp. was obtained at 300 g AI ha^?1 with 1200 g ha^?1 ‘Dobanol’ 25–7; with even better control at higher rates of application of both components. This improvement in performance was accompanied by a higher risk of crop phytotoxicity than observed with the ‘Commando’/‘Swirl’ mixtures. Symptoms initially were scorch and subsequently growth depression, particularly of tillers. None of the mean values in the six ‘efficacy’ trials reached commercially unacceptable levels, but in a further six ‘crop effects’ trials (three wheat, three barley), in which double rates were applied, the levels of phytotoxicity did become unacceptable and subsequently reduced grain yields. In contrast, two barley ‘crop effects’ trials gave yields higher than the control plots, possibly through the effects of reducing stem length and lodging thereby enabling more efficient harvesting. Nevertheless, there were rates of application of flamprop-M-isopropyl in the range 300–400 g ha^?1 with ratios of ‘Dobanol’ 25-7 in the range 2:1 to 4:1 that would achieve high levels of control of Avena spp. without undue risk of crop phytotoxicity and further trials are planned to support this new adjuvant system.     

Usefulness of aerobic microbial culture and cytologic evaluation of corneal specimens in the diagnosis of infectious ulcerative keratitis in animals

OBJECTIVE: To determine the diagnostic value of aerobic microbial culture and cytologic evaluation of corneal specimens in the diagnosis of infectious ulcerative keratitis (IUK). DESIGN: Prospective study. ANIMALS: 48 animals (26 dogs, 13 horses, 7 cats, 1 bird, and 1 llama) with corneal ulcers. PROCEDURE: Scrapings from corneal ulcers were examined cytologically. Corneal swab specimens were submitted for microbial culture. Animals were grouped according to whether they had been receiving antimicrobials at the time of admission. RESULTS: Of the 38 animals receiving antimicrobials, 19 had positive results for IUK on cytologic evaluation, 20 on microbial culture, and 26 on cytologic evaluation, microbial culture, or both. Of the 10 animals not receiving antimicrobials at the time of admission, 7 had positive results for IUK on cytologic evaluation, and 9 had positive results on microbial culture. In this group of 10 animals, additional animals with IUK were not identified on the basis of cytologic evaluation alone. When all 48 animals were considered irrespective of antimicrobial treatment, 26 and 29 had positive results for IUK on cytologic evaluation and microbial culture, respectively, whereas IUK was confirmed in 35 animals on the basis of cytologic evaluation, microbial culture results, or both. CONCLUSIONS AND CLINICAL RELEVANCE: Microbial culture and cytologic evaluation of corneal specimens maximizes identification of IUK, especially in animals receiving antimicrobial treatment. Because of serious consequences of untreated IUK, we recommend that both diagnostic tests be used to tailor treatment and reduce risk of vision impairment in animals.