苜蓿愈伤组织原生质体游离与培养

对阿尔冈金苜蓿(Medicago sativa L. cv. Algonguin)原生质体游离和培养的最佳条件进行研究,建立其细胞融合原生质体杂交体系。以下胚轴诱导的愈伤组织为材料,研究了愈伤组织继代培养时间、酶液组合、酶解时间及酶液渗透压对原生质体游离的影响,并探讨了愈伤组织原生质体在KM8P培养基上的分裂情况。结果表明:继代12d的愈伤组织游离性最好,最佳酶液组合为2%纤维素酶+0.5%果胶+1%半纤维素酶、最佳酶解时间为12h、最佳酶液渗透压为0.55mol/L;最适宜培养苜蓿愈伤组织原生质体的培养基为KM8P+2.0mg/L2,4-D+1.0mg/LNAA。     

甘农4号紫花苜蓿愈伤组织原生质体游离条件的研究

以甘农4号紫花苜蓿下胚轴诱导所得愈伤组织为材料,研究酶液组合,渗透压调节物甘露醇浓度,酶解时间,愈伤组织继代时间及预处理措施对原生质体游离效果的影响。结果表明:当酶液组合为2%纤维素酶+0.5%果胶酶+0.3%半纤维素酶,渗透压为0.6mol/L甘露醇,愈伤组织继代培养时间为12d,预处理措施为在CPW-11溶液中预质壁分离1h,酶解时间10～12h时,游离出的原生质体产量最高,可达3.34×106个/g,原生质体活力高达82.8%。     

扁蓿豆愈伤组织原生质体分离条件的研究

以扁蓿豆(Melilotoides ruthenica(L.)Sojak)下胚轴愈伤组织为材料,研究酶液组合、酶解时间、酶液渗透压、继代培养时间及预处理措施对其原生质体分离效果的影响,以期确定能够酶解出高数量且高活力的原生质体的酶解条件。结果表明:获得有活力原生质体的最佳酶液组合为2%纤维素酶+0.5%果胶酶+0.3%半纤维素酶、酶解时间为12 h、酶液渗透压即甘露醇浓度为0.55 mol·L^-1,愈伤培养天数为10~12 d、预处理措施为黑暗24 h。     

陇东野生紫花苜蓿愈伤组织原生质体游离条件的筛选

以陇东野生紫花苜蓿子叶诱导的愈伤组织为材料,研究了酶解时间、酶液组合、酶液中甘露醇浓度、预处理措施及愈伤组织继代培养时间对其原生质体游离效果的影响.结果表明:当酶解时间为10h、酶液组合为2%纤维素酶+0.5%果胶酶+0.3%离析酶+0.3%半纤维素酶+0.3%崩溃酶、甘露醇浓度为0.60mol/L、预处理措施为CPW-10溶液中浸泡1h、愈伤继代培养天数为12～14d时,游离出的原生质体产量最高,可达19.5×105个/g,存活率高达89.7%.     

桑树原生质体分离条件的优化试验

为了分离获取高产量、高活力的桑树原生质体,分别以桑树的组培苗细切叶片、胚性悬浮培养细胞、粗切愈伤组织和细切愈伤组织等为材料,采用正交试验和单因素试验的方法对桑树原生质体酶解分离条件中的分离酶液组合、渗透压调节剂、酶解温度、酶解时间等因素进行优化。最佳分离酶液组合为100 U/mL纤维素酶R-10+150 U/mL果胶酶Y-23+6 U/mL离析酶R-10+细胞-原生质体洗液(1 480 mg/L CaCl2.2H2O,27.2 mg/L KH2PO4),以0.6 mol/L甘露醇为渗透压调节剂,在28℃酶解温度条件下酶解6 h,用桑树胚性悬浮培养细胞作分离材料可获得7.8×106个/g的原生质体产量,且原生质体活力达91.4%。研究结果显示,选择合适的分离材料以及酶解分离条件是高效获取高活力桑树原生质体的关键。     

草地早熟禾午夜Ⅱ号原生质体培养及植株再生

对草地早熟禾(Poa Pratensis L.)品种-午夜Ⅱ号进行了原生质体培养及其植株再生的研究。以午夜Ⅱ号成熟种子诱导的松软胚性愈伤组织为材料,利用酶解法分离出大量有活力的原生质体。原生质体经培养,持续分裂形成愈伤组织,并分化出再生绿苗和白化苗。研究了不同酶液组成和酶解时间对原生质体游离的影响。结果表明:采用继代培养8-10 d的胚性愈伤组织在1.0%纤维素酶+1.0%离析酶+0.5%果胶酶+0.3%崩溃酶条件下,酶解14-16 h可得到产量和活力较高的原生质体。原生质体以2.0×10⁵-5.0×10⁵个/mL的植板密度,采用液体浅层法在附加1.0 mg/L 2,4-D、0.3 mg/L 6-BA、100 mg/L水解酪蛋白、100 mg/L水解乳蛋白、1.0%蔗糖、0.4 mol/L甘露醇的KM₈P培养基中培养,可形成较小的愈伤组织。愈伤组织经过增殖在分化培养基上(MS+0.5mg/LNAA+2.0 mg/L 6-BA)可诱导芽、根的形成,再生出完整植株。     

紫花苜蓿原生质体游离条件的研究

以选育的XN-1型紫花苜蓿种子发育5～7d的子叶、下胚轴及其诱导的愈伤组织和分化的无菌苗的叶片为材料,研究了不同的酶种类及组合、酶液浓度、酶解时间以及离心转速和时间等因子对其原生质体游离的影响.结果表明:子叶及第2次继代的愈伤组织为最好的游离材料;最佳酶液组合和酶解时间分别为纤维素酶2% 果胶酶0.5%和8h;最佳离心转速和离心时间分别为500r/min和6min.     

甘肃野生草地早熟禾原生质体分离与培养研究

建立高效的原生质体再生体系是通过原生质体融合技术培育草地早熟禾(Poa pratensis)新品种的重要前提。以甘肃陇西野生草地早熟禾(LX)和定西野生草地早熟禾(DX)胚性愈伤组织为材料,探索其原生质体游离和培养条件。结果表明,LX和DX原生质体分离的最佳酶液组合为1.5%纤维素酶R-10+0.5%果胶酶Y-23+1.0%离析酶R-10+0.3%崩溃酶;酶解时间为16 h;LX原生质体最适宜的甘露醇浓度为0.6 mol·L-1,而DX原生质体的最适甘露醇浓度为0.5 mol·L-1。LX原生质体的产量最高可达6.59×106个·g-1,DX可达5.95×106个·g-1。DX原生质体培养的最适密度为3.0×105个·m L-1,最适2,4-D浓度为1.0 mg·L-1,此时,DX原生质体再生细胞的分裂频率可达9.56%,植板率为4.62%。     

百脉根原生质体的分离和酶解条件的研究

分别以百脉根里奥(Lotus corniculatus L.cv.Leon)无菌苗子叶及由下胚轴和子叶形成的胚性愈伤组织为供体材料,进行原生质体游离,研究不同酶解条件对原生质体分离的影响,以期为进一步开展属间体细胞杂交及培育新品种奠定研究基础。结果表明:子叶和愈伤组织原生质体分离最适的酶液组合均为2%纤维素酶+0.5%果胶酶+0.3%半纤维素酶、以及0.55 mol·L^-1甘露醇,产量及活力分别可达4.13×10⁶个·g^-1,65%和3.8×10⁶个·g^-1,67.4%;最佳酶解时间分别为6 h和12 h。预培养8~10 d,CPW-10溶液中预质壁分离1 h或预暗培养1 d等预处理措施可显著提高愈伤组织原生质体的产量及活力(P<0.01)。     

桑树内生真菌Macrophomina phaseolina MOD-1原生质体制备条件的优化试验

为了应用原生质体技术改良桑树内生产油真菌Macrophomina phaseolina MOD-1的产油性能,对影响MOD-1原生质体制备的酶液组分、菌龄、酶解时间、酶解温度、渗透压稳定剂种类等因素进行优化。通过正交试验得到最佳酶液组合为:20mg/mL纤维素酶+20 mg/mL溶菌酶+5 mg/mL蜗牛酶。在此基础上应用响应面分析法获得影响MOD-1原生质体产量的显著因素有菌龄和渗透压稳定剂浓度,且均产生负效应。以此作为中心点进一步优化原生质体制备条件为:培养44.58 h的MOD-1菌丝,用0.66 mol/L KCl作渗透压稳定剂,30℃条件下酶解4 h。在此优化条件下,获得的MOD-1菌株原生质体产量可达7.06×106个/mL,优化条件具有实用价值。     

癸氧喹酯干混悬剂含量测定的改进

采用高效液相色谱法测定癸氧喹酯干混悬剂的含量,在2-250μg/mL范围内,峰面积的常用对数与进样量浓度的常用对数呈良好的线性关系,R^2=1(n=5),平均回收率为99.24%~99.51%,RSD在0.05%~0.28%。此方法分析时间短,样品前处理简便、定量结果准确,重现性好,结果满意,为其质量控制提供了依据。     

商会自治的基石——商会自治规范研究

在现代法律秩序中,商会自治规范是制定法的基础和必要的补充,甚至在某些方面替代了制定法;商会自治规范主要包括商会组织规范、行为规范、惩罚规范以及争端解决规范等;其效力仅及于其内部成员;商会自治规范和制定法之间存在冲突,但也存在整合的基础。     

猪毛色遗传的研究进展

本文概述了猪的毛色类型、猪的毛色遗传模式,着重综述了猪毛色基因分子基础的研究进展,指出存在问题并就未来发展方向做了思考。     

中华人民共和国农业部公告 第267号

为贯彻落实《兽药生产质量管理规范》(简称《兽药GMP》),进一步推动兽药GMP实施进程,我部制定了《兽药生产质量管理规范检查验收办法》,现予公告。本公告自2003年6月1日起施行。附件:兽药生产质量管理规范检查验收办法二○○三年四月十日第一章 总则 第一条 为推动《兽药生产质量管理规范》(以下简称兽药GMP)的实施,规范兽药GMP检查验收工作,制定本办法。 第二条 农业部负责全国兽药GMP管理和检查验收工作;负责制修订兽药GMP检查验收管理规定;负责兽药GMP检查员队伍建设和监督管理工作,负责国际兽药贸易中GMP互认工作。 …     

鸡败血支原体垂直传播模式及其阻断方法

以国际标准强毒Ｒ株人工感染非免疫产蛋鸡,定时扑杀,分别从鼻窦、眶下孔、气管、肺、气囊、卵巢和输卵管分离ＭＧ,并收集感染鸡所产蛋分离ＭＧ。结果表明,人工感染４８小时后上、下呼吸道及肺已被全面感染,９６小时气囊已被感染,１２０小时输卵管已能分离到ＭＧ,卵巢始终分离不到ＭＧ。人工感染鸡自１４４小时便能在其所产蛋中分离出ＭＧ。药物治疗能在７２小时内消除感染,油乳剂苗则需２４天后逐渐降低蛋内ＭＧ分离率,药物卵内注射、种蛋药浴、高温处理均能杀死卵内ＭＧ,但以研制的种蛋浸泡剂药浴效果为最好。     

Comparison of 2 methods of centesis of the bursa of the biceps brachii tendon of horses

REASONS FOR PERFORMING STUDY: Centesis of the bicipital bursa using an 8.9 cm long spinal needle has been reported but the alternative of employing a 3.8 cm long hypodermic needle requires validation. OBJECTIVE: To compare the efficacy of 2 different methods of centesis of the bicipital bursa and to evaluate the usefulness of ultrasonographic imaging to determine the location of solution administered when centesis of the bursa is attempted. METHODS: For Trial 1, 6 clinicians, who had no previous experience of centesis of the bicipital bursa, attempted to inject a solution composed of an aqueous radiopaque contrast medium and physiological saline solution (PSS) into the bicipital bursae of 2/12 horses using the previously described distal approach to inject one bursa and a proximal approach to inject the contralateral bursa. The bicipital tendon and bursa were examined ultrasonographically before and after injection; and both shoulders were examined radiographically to identify the location of the medium. In Trial 2, another 6 clinicians, also with no previous experience of centesis, repeated Trial 1, using 6 horses, but the radiopaque contrast medium was mixed with air instead of PSS. RESULTS: Accuracy of centesis using the proximal approach was 39% and that of the distal approach 28%. Ultrasonographic examination of the shoulder allowed the location of solution and air to be accurately predicted in all 12 shoulders examined. CONCLUSIONS: Clinicians who have had no previous experience performing centesis of the bicipital bursa are unlikely to be successful in centesis using either approach. Radiographic examination after injecting a radiopaque contrast medium may be necessary to assess the success of centesis especially if bursal fluid is not obtained during centesis. Injecting air along with the radiopaque contrast medium provides more accurate ultrasonographic confirmation of centesis and better radiographic definition than does injection without air.     

不同样本商品蜂蜜中硒含量的测定

用硝酸和高氯酸消化蜂蜜,使硒游离出来,在微酸性环境下,硒和2,3-二氨基萘(DAN)生成有较强荧光的物质,用环己烷萃取,在激发波长378nm,荧光波长518nm处测定其荧光强度。蜂蜜中硒含量范围:0.10～0.82μg/g。表明:蜂蜜应视为天然富硒营养品。     

乳酸杆菌益生作用机制的研究进展

乳酸杆菌作为益生菌广泛用于人和动物。本文综述了乳酸杆菌改善宿主健康的机制。乳酸杆菌可通过产生抗菌物质如乳酸、过氧化氢、细菌素,或者通过竞争营养或肠道黏附位点来抑制致病菌;通过诱导黏附素的分泌或阻止细胞凋亡而增强肠道的屏障功能,从而保护肠道。文章重点讨论了乳酸杆菌表面成分（表面蛋白、脂磷壁酸和肽聚糖）与肠道受体（Ｃ型凝集素受体、Ｔｏｌｌ样受体和 Ｎｏｄ样受体）,阐述了他们结合后启动免疫调节信号,调控肠道免疫功能以发挥改善健康作用的机制。     

Effects of size of ingestively masticated fragments of plant tissues on kinetics of digestion of NDF

Ingestively masticated fragments were collected and sized via sieving. Different sizes of esophageal masticate and ruminal digesta fragments, and ground fragments of larger masticated pieces were incubated in vitro, and undigested NDF remaining at intervals of up to 168 h of incubation was determined. The ruminal age-dependent time delay (tau) for onset of digestion of NDF was positively correlated (P < 0.004) with the mean sieve aperture estimated to retain 50% of the fragments between successive sieve apertures (MRA). Degradation rate of potentially degradable NDF (PDF) and level of indigestible NDF were not related (P > 0.10) to MRA of masticated and ground fragments. Estimates of tau were positively related to MRA, with slopes of bermudagrass < corn silage < ruminal fragments of corn silage. It was concluded that fragment size-, and consequently, ruminal age-dependent onset of PDF degradation of a mixture of different fragment sizes results in an age-dependent rate of degradation of the more rapidly degrading of two subentities of PDF. Models are proposed that assume a tau before onset of simultaneous degradation of PDF from two pools characterized as having gamma-modeled age-dependency and age-constant rates. The ruminal age-dependent pool seems to be associated with the faster-degrading pool, and its rate parameter increases with range in MRA in the population of fragments. Conceptually, the ruminal age-dependent rate parameter for PDF degradation seems to represent a composite of several effects: 1) effects of the size-dependent tau; 2) range in MRA of the population of ingestively masticated fragments; and 3) subentities of PDF that degrade via more rapid age-dependent rates compared with subentities of PDF that degrade via age-constant rates. The estimated fractional rates of ruminative comminution of ingestively masticated fragments (0.060 to 0.075/h) were of a magnitude similar to the mean fractional rates of PDF digestion (0.030 to 0.085/h), which implies that ruminative comminution may be first-limiting to fractional rate of PDF digestion. The in vivo roles of ingestive and ruminative mastication of fragments on PDF degradation must be considered in any kinetic system for estimating PDF digestion in the rumen. These results and others in the literature suggest that the rate of surface area exposure rather than intrinsic chemical attributes of PDF may be first-limiting to degradation rate of PDF in vivo.