不同培养基对禾草内生真菌Epichloё生长与产孢的影响

本研究以分离自西藏早熟禾(Poa tibetica)、甘肃臭草(Melica przewalskvi)和麦宾草(Elymus tangutorum)的6个内生真菌菌株(分别为西藏早熟禾内生真菌Ep-003和Ep-004,甘肃臭草内生真菌Em-014和Em-016,麦宾草内生真菌Ee-001和Ee-003)为研究对象,分别在马铃薯葡萄糖琼脂(PDA)、麦秆煎液琼脂(WSA)、玉米粉琼脂(CMA)和水琼脂(WA)4种培养基上培养4周后,观测菌落生长速率、菌落形态特征、产孢量和孢子特性。结果表明,同一菌株在不同培养基上的生长速率一般为PDA和CMA上的最大,WSA和WA上最小;不同菌株在同一培养基上,西藏早熟禾的Ep-003菌株生长最快,在PDA上直径最大,为46.42 mm,麦宾草内生真菌次之,甘肃臭草的Em-014菌株生长最慢,在WA上直径最小,为7.07 mm;在4种培养基上,菌株的产孢量有显著差异(P0.05),分别为Ep-003只在WA上产孢,其他5种菌株在WA和WSA培养基上产孢较多,在PDA和CMA上产孢较少;Em-016和Ep-004在4种培养基上的孢子长度无显著差异(P0.05),Em-014在WSA上的孢子长最小,为3.77μm,其他菌株无显著差异(P0.05),Ee-003在PDA上孢子长最长,为8.43μm,在其他3种培养基上无显著差异(P0.05);Ep-004、Em-016和Ee-003在4种培养基上孢子梗长度均无显著差异(P0.05),Em-014在WA上孢子梗最长,为16.74μm,在WSA上最短,为14.09μm,Ee-001在CMA上孢子梗最长,为18.59μm,在WSA上最短,为13.40μm。     

不同培养基对禾草内生真菌Epichlo(e)生长与产孢的影响

本研究以分离自西藏早熟禾(Poa tibetica)、甘肃臭草(Melica przewalskvi)和麦宾草(Elymus tangutorum)的6个内生真菌菌株(分别为西藏早熟禾内生真菌Ep-003和Ep-004,甘肃臭草内生真菌Em-014和Em-016,麦宾草内生真菌Ee-001和Ee-003)为研究对象,分别在马铃薯葡萄糖琼脂(PDA)、麦秆煎液琼脂(WSA)、玉米粉琼脂(CMA)和水琼脂(WA)4种培养基上培养4周后,观测菌落生长速率、菌落形态特征、产孢量和孢子特性.结果表明,同一菌株在不同培养基上的生长速率一般为PDA和CMA上的最大,WSA和WA上最小;不同菌株在同一培养基上,西藏早熟禾的Ep-003菌株生长最快,在PDA上直径最大,为46.42 mm,麦宾草内生真菌次之,甘肃臭草的Em-014菌株生长最慢,在WA上直径最小,为7.07 mm;在4种培养基上,菌株的产孢量有显著差异(P＜0.05),分别为Ep-003只在WA上产孢,其他5种菌株在WA和WSA培养基上产孢较多,在PDA和CMA上产孢较少;Em-016和Ep-004在4种培养基上的孢子长度无显著差异(P＞0.05),Em-014在WSA上的孢子长最小,为3.77 μm,其他菌株无显著差异(P＞0.05),Ee-003在PDA上孢子长最长,为8.43 μm,在其他3种培养基上无显著差异(P＞0.05);Ep-004、Em-016和Ee-003在4种培养基上孢子梗长度均无显著差异(P＞0.05),Em-014在WA上孢子梗最长,为16.74 μm,在WSA上最短,为14.09 μm,Ee-001在CMA上孢子梗最长,为18.59 μm,在WSA上最短,为13.40 μm.     

苜蓿假盘菌的生物学特性研究

摘要：本试验研究了温度、水分、pH值、光照、碳源、氮源以及无机盐对苜蓿假盘菌(Pseudopeziza medicaginis)宁夏菌株生物学特性的影响。结果表明,苜蓿假盘菌宁夏菌株在5～30 ℃、pH值4.0～10.0均可生长,最适温度为20～25 ℃,最适pH值为5.0～6.5,该菌随光照强度的增加生长受到抑制,能利用固体培养基上不同的碳源,对葡萄糖和麦芽糖的利用较好。除尿素抑制病菌的生长外,该菌能不同程度利用无机氮源和有机氮源,菌落直径和生长速率基本一致,差异不明显,但菌落数差别较大,无机氮源优于有机氮源。供试的4种盐溶液除KH2PO4没有菌生长外,其余3种生长量基本一致,但4种盐溶液下生长的菌落均不产生子囊盘。叶片表面的游离水及游离水保持的时间可影响孢子的萌发,且叶表游离水保持3 h以上,孢子才能萌发。     

三种镰孢菌对不同苜蓿品种致病性的比较研究

为了进一步了解根腐病病原菌的生物学特性以及不同病原菌对苜蓿的致病性,本研究以苜蓿根腐病病原BF1(Fusarium equiseti)、BF28(F.oxysporum)、BF36-2(F.solani)三种镰孢菌菌株为试验材料,观测菌落的外观特征与产孢量,并对WL343、金皇后、阿尔冈金、WL525四个苜蓿品种以菌液和菌饼两种方式接种,比较不同菌株对苜蓿的致病性。结果表明,菌液接种方式,苜蓿感病较快,感病程度较重。不同菌株在苜蓿根系的定植能力不同,BF1菌丝在根系附近聚集;而BF28及BF36-2菌丝则均匀分布于培养基表面。菌饼接种方式,不同菌株对苜蓿的致病性存在较大差异,产孢能力强的菌株致病力强。BF28对苜蓿的致病性最强,BF36-2次之,而BF1的致病性较弱。通过对感病植株进行解剖构造与致病过程观察,病菌以厚垣孢子入侵根系及叶片表皮细胞,以菌丝体及孢子形式在细胞间隙扩展,并向植物组织中释放细胞壁溶解物质,植物细胞壁溶解,根髓部腐烂中空,根颈部缢缩,叶部出现溃烂斑,植株的生长受阻。     

箭筈豌豆炭疽病病原菌分离鉴定

箭筈豌豆炭疽病是一类重要的真菌性病害。采用形态学特征观察结合基于内转录间隔区(ITS)、肌动蛋白(ACT)、几丁质酶(CHS1)和3-磷酸甘油醛脱氢酶(GADPH) 4个基因序列的多基因系统发育分析方法来鉴定甘肃夏河箭筈豌豆炭疽病的病原菌。结果表明,菌株WQ在马铃薯葡萄糖琼脂(PDA)培养基上的菌落平坦,正面为深灰色-橄榄色至橄榄色-黑色,边缘菌丝白色,背面灰色-橄榄色至铁灰色。分生孢子单胞透明,大小为(17.5～25.0)μm×(3.75～5.00)μm,略呈镰刀形。菌株WQ与编号为CBS 128.57的菌株同源性最高,在70%水平上聚为一支。病原菌可以通过伤口侵染箭筈豌豆叶片。确定箭筈豌豆炭疽病病原菌为菠菜炭疽菌,这是箭筈豌豆菠菜炭疽菌在中国的首次报道。寄主范围测定结果表明,病原菌对红豆草和燕麦均具有强致病性,对4个品种的箭筈豌豆具有中等致病性,对苜蓿和三叶草的致病性最低,而对黑麦草无致病性。     

中国新疆苜蓿野生种群秋眠性的研究

以美国9个秋眠性标准对照品种为参照，按Barnes(1991)方法测定了中国新疆野生苜蓿种群117份、伊朗及其它国家35份材料的秋眠性。结果表明，中国新疆北疆苜蓿野生种群没有半秋眠性种质，均为秋眠性种质，而南疆苜蓿则有秋眠性等级为4和5的种群多份；来自伊朗等国的材料则具有秋眠、半秋眠、非秋眠类型材料，对中国南疆、北疆野生种群秋眠性差异大的原因进行了探讨。     

中国新疆苜蓿野生种群秋眠性的研究

以美国9个秋眠性标准对照品种为参照，按Barnes（1991）方法测定了中国新疆野生苜蓿种群117份、伊朗及其它国家35份材料的秋眠性。结果表明，中国新疆北疆苜蓿野生种群没有半秋眠性种质，均为秋眠性种质，而南疆苜蓿则有秋眠性等级为4和5的种群多份；来自伊朗等国的材料则具有秋眠、半秋眠、非秋眠类型材料，对中国南疆、北疆野生种群秋眠性差异大的原因进行了探讨。     

溶磷菌在5种不同培养基中溶解磷矿粉的性能比较

通过小麦、苜蓿根际分离的4株溶磷菌(Lx81、Dm84、Jm92、Lx191)接种于PKO、SP、NBRIY、NBRIP、NBRIYP等5种培养基中测定菌落直径、溶磷圈大小、培养液有效磷含量、pH值及总有机酸含量的方法,研究溶磷菌在不同培养基上的生长情况和溶磷特性。结果表明:Lx81和Lx191在PKO培养基上的菌落直径最大,Dm84和Jm92在NBRIY培养基上的菌落直径最大;4菌株在NBRIP培养基中的溶磷量、总有机酸含量、pH下降值均显著高于其他培养基中的值。为了得到更多、更有效的溶磷菌株,在以后的工作中建议采用NBRIP培养基。     

马玲薯葡萄糖培养基在延胡索酸泰妙菌素菌种选育中的应用

采用涂布法将延胡索酸泰妙菌素生产菌种TMU10—444分离于不同平板培养基上（生产用麦芽浸膏培养基和马铃薯葡萄糖培养基）,置恒温恒湿培养箱中培养成熟后,挑取单菌落、初筛、复筛,采用高效液相色谱（HPLC）法测定效价。结果表明,两种不同平板培养基上的菌落生长有差异,马铃薯葡萄糖培养基上的菌落比对照培养基上的菌落明显长得快;对马铃薯葡萄糖培养基选育的菌株进行了筛选,最终获得4支高于对照生产菌株,分别是2107、2115、2165和2183。此结论为延胡索酸泰妙菌素产生菌菌种选育提供了一种新方法。     

尖孢镰刀菌生物学特性及杀菌剂毒力测定

为了明确苜蓿根腐病致病菌尖孢镰刀菌的生物学特性和最佳杀菌剂,本试验采用不同培养基、温度、pH、光照和碳氮源的条件培养尖孢镰刀菌,测定菌落生长速率和孢子萌发率;用4种杀菌剂与PDA培养基混合制成含药培养基培养尖孢镰刀菌,测定菌落生长速率和抑制率。结果表明:适宜培养尖孢镰刀菌培养基有PSA、SNA和YMM;适宜生长温度为20～30℃、pH为7～8;黑暗条件适于生长。其中菌丝生长和孢子萌发的最适温度分别为25℃和20℃,最适pH分别为7和5,最适碳源分别为可溶性淀粉和麦芽糖,最适氮源分别为尿素和蛋白胨,菌丝和孢子的致死温度分别为52℃(10min)和50℃(10min)。在供试的4种杀菌剂中,75%百菌清抑制效果最好,在稀释500倍时抑制率为89%,EC_(50)为4.5279mg/L。     

桑枝枯菌核病病原菌的生物学特性研究

从发病桑树上分离到桑枝枯菌核病病原菌的纯菌种 ,并对该菌的培养性状和生物学特性进行了调查和研究。结果表明 :桑枝枯菌核病病原菌适于在PDA培养基和桑叶汁培养基上生长 ,其生长温度范围为 8～ 30℃ ,最适生长温度为 2 0～ 2 5℃ ,生长pH值范围为 1～ 12 ,最适生长pH值为 5～ 8,菌丝与菌核的生长情况大致相同 ;菌核在适宜的温湿度条件下能够产生子囊盘和子囊孢子 ,子囊孢子椭圆形 ,大小 7～ 14 μm× 3～ 7μm。     

ISOLATION OF MYCOPLASMA HYOPNEUMONIAE AND ITS ASSOCIATION WITH PNEUMONIA OF PIGS IN AUSTRALIA

Nine strains of mycoplasmas were isolated from the lungs of 5 pigs with clinical signs of naturally acquired enzootic pneumonia. Mycoplasma hyopneumoniae was isolated from the lungs of 1 pig and M. hyorhinis from the lungs of 4. An unidentified mycoplasma, which utilized arginine, grew rapidly in broth culture and produced centred colonies on solid medium, was isolated from the lungs of 4 pigs. The pathogenicity of the isolated strain of M. hyopneumoniae was determined by inoculation of pigs from an enzootic pneumonia-free herd. Enzootic pneumonia was produced in the lungs of all 5 pigs inoculated intranasally and intratracheally with broth cultures of the organism isolatied by limit dilution techniques. Enzootic pneumonia was produced in 3 of 6 pigs inoculated intranasally and intratracheally with M. hyopneumoniae purified by the passage of colonies on agar blocks. M. hyopneumoniae was isolated in pure culture from the lungs of all pigs with induced pneumonic lesions.     

破碎法提取家蚕微孢子虫孢子核酸的PCR检测灵敏度试验

蚕微粒子病检测技术是该病防治中的一项重要技术,本文在通过破碎法提高模板核酸获取量的基础上,对纯化家蚕微粒子虫孢子的PCR检测灵敏度进行了测试。提取核酸-模板稀释测试12对引物的灵敏度中有4对引物达到0．05粒（10^2粒/mL）、4对引物5粒（10^4粒/mL）、1对引物50粒（10^5粒/mL）、2对引物500粒（1...     

青海部分牧区牦牛酸奶中乳杆菌的分离与鉴定

利用MRS培养基,对青海部分牧区收集的15份牦牛酸奶,进行了乳杆菌的分离,对分离得到的9株疑似乳杆菌,对其中的3株进行生物学鉴定。结果:分离株均为兼性厌氧菌,在MRS培养基上,菌落较大、透明、灰白色,镜检为杆状细菌,革兰氏染色阳性,过氧化氢酶试验为阴性,乳糖发酵阳性,牛乳发酵试验阳性,符合乳杆菌的特性。     

Use of isolated infected spores to determine the sporocidal efficacy of two commercial antifungal rinses against Microsporum canis

In this study, isolated infective Microsporum canis spores were used in an in vitro test model to compare the sporocidal activity of two commercial topical antifungal rinses. The two commercial test solutions used in the study were a lime sulphur solution 97.8% (LymDyp) and miconazole base 5.2%/chlorhexidine gluconate 5.9% mixture (Malaseb Concentrate Rinse). Water and household bleach were used as controls. Isolated infective spores were harvested from infected hairs and 500 microL of the spore suspension was incubated with an equal volume of dilutions of lime sulphur or the miconazole/chlorhexidine gluconate combination for 5 min and 4 h followed by fungal culture. There were too many to count colonies on the water control plates. Lime sulphur was 100% sporocidal at all test dilutions at both times. Miconazole/chlorhexidine gluconate was 100% sporocidal at all but the 1 : 128 dilution after 5 min of incubation and 100% sporocidal when incubated with spores for 4 h. It is not known if the two products have similar antifungal activity against infective spores on or within hairs; however, based on the findings of this study there is good evidence to recommend either rinse as an adjuvant topical therapy in a dermatophyte treatment and control program.     

Bovine oocytes in secondary follicles grow in medium containing bovine plasma after vitrification

There has been no culture system that supports the growth of bovine oocytes for more than 2 weeks. In the present study, bovine secondary follicles were cultured for 4 weeks, and the effects of supplemented protein components and FSH in the culture medium on the growth of the oocytes were examined. The effect of vitrification of secondary follicles on the subsequent oocyte growth was also examined. Secondary follicles (150 to 200 μm in diameter) containing growing oocytes (approximately 60 μm in diameter) were dissected from ovaries and cultured in a medium supplemented with FSH (0, 25 or 50 ng/ml) and one of the following four kinds of protein components: bovine serum albumin (BSA), bovine plasma (BPL), fetal calf serum (FCS) and bovine follicular fluid (BFF). In BSA- and BPL-supplemented media with 0 or 25 ng/ml FSH, more than 50% of follicles showed no degenerative signs during culture, and oocytes significantly increased in size after 4 weeks (P<0.05). Higher percentages of granulosa cell-enclosed oocytes were recovered from the follicles cultured in BPL-supplemented media with 0 and 25 ng/ml FSH, and the oocytes grew to 90 μm or more in diameter. In FCS- and BFF-supplemented media, FSH increased the numbers of degenerating follicles. Next, vitrified-warmed secondary follicles were cultured in BPL-supplemented medium. One third of the follicles showed no degenerative signs, and the oocytes increased in diameter to 88.8 ± 3.1 μm after 4 weeks of culture. These results suggest that a BPL-supplemented medium supports oocyte growth in bovine secondary follicles for 4 weeks, even after vitrification and warming of the follicles.     

布氏锥虫伊氏亚种体外培养的研究——Ⅲ.集落的形成及其生物学意义

以无细胞培养系统培养布氏锥虫伊氏亚种(Trypanosma brucei subsp.evansi,简称伊氏锥虫),可见其有规律地形成集落.在每日混匀换液后8h开始虫体向局部聚集,形成集落;随着聚集过程的进展,集落范围缩小,虫密度增高,与周围境界愈加明显.12h集落大小趋于稳定,而虫密度仍持续上升至18h其表现为集落内虫体向多层立体分布发展.虫体不附着于培养器皿,运动活泼.在集落虫密度不断增高的同时,集落内单个虫体逐渐减少,而由分裂态虫体形成的小虫簇相应增多.集落实际上是旺盛分裂增殖的虫体群落.从培养8h起,每2h把培养物混匀1次,其前12h虫密度增长情况与始终静置培养的培养物基本一致,但12h后不再增殖,而后者则持续增殖至18h致使前者的最高虫密度仅为后者的约3/4.结果说明,集落的形成和保持有助于生长繁殖,是伊氏锥虫固有的生物学特性.本文还讨论了其形成机制和生物学意义.     

Use of a commercial methylcellulose medium with and without recombinant bovine granulocyte colony-stimulating factor for culturing bovine bone marrow cells

A commercial methylcellulose culture medium, with and without the addition of recombinant bovine granulocyte colony-stimulating factor (rbG-CSF), was utilized for culturing bovine bone marrow cells in a colony-forming unit assay. Bone marrow mononuclear cells were isolated and cultured in a commercial methylcellulose-based medium containing several recombinant human cytokines. Cultures were prepared with and without 100 ng/mL of rbG-CSF. The size and mean number of colonies per plate from culture days 3 to 9 were compared. We concluded that bovine bone marrow colony growth was supported by this culture medium. The addition of rbG-CSF yielded larger and more numerous colonies. There were significantly more colonies on day 3 (P < 0.001), day 4 (P < 0.001), and day 5 (P = 0.03) with rbG-CSF. Both culture media had the highest colony counts on day 5.     

Analysis on the Genetic Diversity of Six Bactrian Camel Populations in China Using Microsatellite Marker

In order to understand the genetic diversity of Bactrian camel and the genetic evolutionary relationships between different populations,the genetic diversity of Alashan camel, Qinghai camel, Nanjiang camel, Beijiang camel, Subei camel and Sunite camel populations in China were analyzed using 10 microsatellite markers. By calculating heterozygosity (H), polymorphism information content (PIC), effective allele (Ne), Shannon information index, the genetic variation within populations were analyzed. By calculating the F-statistics, gene flow, genetic differentiation coefficient and genetic distance to analyze the genetic relationship between populations. The results showed that 89 alleles were tested at the 10 microsatellite loci,8.9 alleles were tested at every locus in average. All loci were medium-highly polymorphic loci (except YWLL08),the average PIC of the Bactrian camel population was between 0.488 to 0.752. The observed heterozygosity (0.355 to 0.448) of 6 Bactrian camel populations was lower than the expected heterozygosity (0.643 to 0.703). Almost all loci Shannon index was greater than 1,and most of loci were in Hardy-Weinberg disequilibrium. The genetic differentiation coefficient between groups (Fst) value was 0.059 and in the low degree of moderately differentiated state. The average Fis values of 6 Bactrian camel populations were positive which suggested 6 Bactrian camel populations had different levels of inbreeding. The standard genetic distance (D_S) and genetic distance T (D_A) clustering analysis showed that the Nanjiang camel and Beijiang camel jointed as one group, the Alxa camel, Qinghai camel, Subei camel, Sunite camel jointed as the other group. Research showed that Chinese Bactrian camel was abundant in genetic diversity, genetic variation within population was larger, and there was a phenomenon of inbreeding. There was a certain gene flow between populations, the differentiation of populations were mainly caused by genetic variation within population. 6 Chinese Bactrian camel populations were divided into two groups.     

Moment of addition of LH to the culture medium improves in vitro survival and development of secondary goat pre-antral follicles

The present study investigated the effects of time of addition of luteinizing hormone (LH) to culture medium on the in vitro development of caprine pre-antral follicles. Pre-antral follicles (≥ 150 μm) were isolated from fragments of the goat ovarian cortex and individually cultured for 18 days in the absence (control) or presence of 100 ng/ml LH, added on days 0, 6 or 12 of culture. Follicular development was assessed based on antral cavity formation, increased follicular diameter as well as follicular and fully grown oocyte (>110 μm) viability. The results showed that after 18 days of culture, the percentage of surviving follicles in the control treatment was significantly lower when compared to other treatments (p < 0.05). There were no significant differences in antrum formation, follicular diameter and oocyte viability. The addition of LH at D6 of culture significantly increased the rates of oocytes ≥ 110 μm and the resumption of meiosis (p < 0.05). In contrast, when LH was added at the onset of culture, only germinal vesicle oocytes were obtained. In conclusion, the moment of addition of LH to the culture medium affects the performance of in vitro culture of caprine pre-antral follicles. The addition of LH to the medium from day 6 of culture onward improved the rates of follicular survival, as well as the ability of oocytes to resume meiosis. However, prolonged exposure to LH (addition at the onset of culture onward) showed detrimental effects for the meiotic resumption.