甜瓜结实花初花节位QTL分析

 以WI998为母本（纯雌株、厚皮网纹甜瓜品系）,TopMark为父本（雄全同株纯合品系）,配置杂交组合,利用单粒传得到171个株系的重组自交系（F₂S₄）群体构建甜瓜SSR分子标记遗传连锁图谱。该连锁图谱包含19个连锁群,覆盖基因组长度为1 414.2 cM,标记间平均距离为10.2 cM。对结实花初花节位开展QTL分析,共检测到9个QTL,分别分布在第1、9、10、11连锁群上,各QTL的LOD值在2.89 ~ 9.42之间,4个QTL贡献率超过10%。位于第9连锁群的QTL Fp9.4贡献率最大,为18.57%（2010秋季LOD = 9.17）;位于第9连锁群的Fp9.3（2010春季8.64%,LOD = 6.44;2010秋季17.99%,LOD = 9.42）,位于第10连锁群的Fp10.1（2010春季3.59%,LOD = 2.89;2010秋季6.20%,LOD = 3.43）在两季的位置都很稳定。获得与结实花初花节位紧密连锁（< 10 cM）的8个特异标记（SSR01737、MU141991、TJ105、GCM206、SSR04910、MU173563、NR52、MU146331）,为进一步开展QTL精细定位提供参考。     

Application of PCR-HRM in the study of relationship between leptin gene promoter polymorphisms and liver cirrhosis

AIM: To analyze the feasibility of PCR-high-resolution melting curve analysis (HRM) for detecting the site mutations of C2549 and G2548 in leptin gene promoter from the patients with liver cirrhosis, and to explore the relevance between mutant genotypes and physiological and biochemical indexes in liver cirrhosis patients. METHODS: Compared with the method of PCR-restriction fragment length polymorphism (RFLP), the present research used the method of PCR-HRM to analyze the site mutations of C2549 and G2548 in leptin gene promoter in control group (n=100) and liver cirrhosis group (n=100). The physiological and biochemical indexes of the patients were also detected and compared. RESULTS: Leptin gene promoter polymorphism was detected using PCR-HRM with effectiveness, high flux and accuracy. Preliminary results showed that the main mutation of the patients with liver cirrhosis was in C2549 site, but not found in G2548 site. Leptin, free leptin index (FLI), fasting insulin (FINS) and insulin resistance index estimated by homeostatic model assessment (HOMA-IR) in liver cirrhosis group were higher than those in control group. Insulin sensitivity index (ISI) and soluble leptin receptor (sOB-R) in liver cirrhosis group were lower than those in control group with significant difference except leptin level. Meanwhile, FLI showed positively correlated with FINS and HOMA-IR (r=0.45, r=0.53, P<0.05), and negatively with ISI (r=-0.34, P<0.05). In the patients with liver cirrhosis, C2549A heterozygous mutation was predominant. The indexes of HOMI-IR, leptin, sOB-R and FLI of C2549A homozygotes and heterozygotes were higher than those of the wildtypes, which showed significant difference except leptin and sOB-R levels (P<0.05). CONCLUSION: PCR-HRM can be more accurate for identifying leptin promoter polymorphism. The increase in the frequency of C2549A mutation may be closely related with liver cirrhosis. Existence of hyperinsulinemia and insulin resistance may be correlated with leptin level in the patients with liver cirrhosis.     

西瓜花叶病毒抗血清制备及其应用

 西瓜花叶病毒(Watermelon mosaic virus, WMV)是危害葫芦科作物的重要病毒。制备特异性强、效价高的抗血清对快速准确检测和鉴定WMV具有重要意义。本研究将WMV外壳蛋白(Coat protein,CP)基因克隆到原核表达载体pEHISTEV获得pEHISTEV-WMV-CP。将pEHISTEV-WMV-CP转化大肠杆菌Rosetta,经IPTG诱导,成功表达出分子量约为36 kDa的蛋白,与预期WMV CP大小一致。切胶回收WMV CP,与等体积弗氏不完全佐剂充分乳化后免疫健康新西兰大白兔。Western blotting结果表明,制备的抗血清与WMV CP有反应,与同属的番木瓜环斑病毒(Papaya ringspot virus,PRSV)、马铃薯Y病毒(Potato virus Y,PVY)、小西葫芦黄花叶病毒(Zucchini yellow mosaic virus, ZYMV)和烟草花叶病毒属的黄瓜绿斑驳花叶病毒(Cucumber green mottle mosaic virus,CGMMV)均无反应。酶联免疫吸附测定(PTA-ELISA)结果表明,本研究制备的WMV抗血清效价为1∶8 192,并且能够检测稀释512倍的病毒汁液。利用该抗血清对田间采集的10个RT-PCR检测为阳性的样品进行检测,全部呈现阳性。本研究利用大肠杆菌表达的CP制备的WMV抗血清具有较高的特异性和灵敏度。研究结果为WMV的快速检测和鉴定奠定了基础。     

从野外昆虫体内分离的5株微孢子虫的生物学特性与分子系统发育研究

从广西蚕区的野外昆虫菜粉蝶(Pieris rapae)、桑尺蠖(Hemerophila atrilineata)、斜纹夜蛾(Spodoptera litura)的体内收集到5株对家蚕有一定食下感染力的微孢子虫,通过生物学性状、分子系统发育研究,为其分类鉴定提供依据。来自野外昆虫的5株微孢子虫均具有Nosema属的生物学特性:显微镜下观察形状均呈长卵圆形,与家蚕微孢子虫(Nosema bombycis,Nb)有较明显差异;在生活史的各发育阶段均为双核,以二分裂方式增殖,最后形成双核成熟孢子;与Nb抗血清均产生阳性凝聚反应;超微结构观察均具双核,极丝圈数在12～14圈之间,极丝倾斜角与Nb有差异。根据5株野外昆虫微孢子虫与其它昆虫微孢子虫基于SSU rRNA基因序列构建的系统发育树,以及基于SSU rRNA和rRNA ITS基因序列进行的相似性和遗传距离分析,初步认为5株野外昆虫微孢子虫与家蚕微孢子虫同属异种。     

基于GIS和景观生态学的西部地区城镇建设用地扩展研究——以宝鸡市中心城区为例

近些年,城镇建设用地的大规模扩张对区域土地资源承载力形成了巨大压力,并导致了大量土地资源的闲置浪费。为了实现土地资源的集约化利用,文中结合GIS和景观生态学的研究方法对宝鸡市1988～2005年中心城区城镇建设用地扩展类型进行定量识别。结果表明:1)近些年,宝鸡市中心城区城镇建设用地扩展经历了缓慢扩展期(1988～2000年)和高速扩展期(2000～2005年)两个阶段;2)在斑块类型水平上,城镇建设用地表现出较好的连通性,并且呈增强趋势;3)外延式扩展是研究期内城镇建设用地扩展的主要类型,扩展比重为外延式>填充式>跳跃式。通过研究为城市空间规划提供了详细而准确的数据,为寻求优良扩张模式奠定了一定的基础。     

狐和貉源大肠埃希菌药敏试验及耐药基因检测

为了解秦皇岛昌黎县狐和貉源致病性大肠埃希菌耐药性及耐药基因的情况,以昌黎县分离的15株狐和貉源致病性大肠埃希菌为试验菌株,采用药敏纸片法对抗菌药物进行药敏试验,并对四环素类耐药基因tetA、tetB、tetC、tetD,氨基糖苷类耐药基因strA-strB、aadA1,磺胺类耐药基因sul1、sul2、sul3进行PCR检测。结果显示,15株大肠埃希菌分离菌株对土霉素、多西环素、头孢拉定、新霉素、青霉素、磺胺间甲氧嘧啶、磺胺二甲氧嘧啶的耐药率均为100%;对阿莫西林、庆大霉素、头孢曲松、阿米卡星、大观霉素、链霉素、复方新诺明、氨苄西林、环丙沙星耐药率分别为93.33%、53.33%、33.33%、73.33%、86.66%、86.66%、86.66%、66.67%和66.67%。9种耐药基因tetA、tetB、tetC、tetD、strA-strB、aadA1、sul1、sul2、sul3的检出率分别为0、46.67%、26.67%、13.33%、40%、100%、53.33%、46.67%和53.33%,其中四环素类耐药基因以tetB的检出率最高,氨基糖苷类耐药基因以aadA1的检出率最高,磺胺类耐药基因的检出率相差不大。说明15株狐和貉源大肠埃希菌耐药谱广,耐药性高,耐药基因流行普遍。     

基于GIS的陕西省酿酒葡萄气候区划

利用陕西省96个县（区、市）气象站及周边省份临近气象站近30 a（1981—2010年）气象观测资料,辅以经订正后的各县气象哨和水文站观测数据,在对国内外葡萄区划指标分析基础上,确定以≥10℃活动积温为一级指标,酿酒葡萄成熟期8—9月水热系数为二级指标,冬季埋土防寒线、9月降水为辅助指标作为陕西省酿酒葡萄气候区划指标。基于GIS技术,利用小网格推算模型对区划指标空间化后,采用主从叠代方法,制作陕西省酿酒葡萄气候区划图。结果表明,陕西省酿酒葡萄种植可分为4个气候区和12个气候亚区,自兴平以东的关中东部海拔900 m以下地区是陕西省种植酿酒葡萄的最优区;对各区酿酒葡萄生产的气候因子进行了评价,提出了相应的生产建议。     

哈茨木霉对水稻恶苗病菌的拮抗作用

PDA平板拮抗试验表明 ,哈茨木霉对水稻恶苗病菌有强烈的拮抗作用 ,其孢子悬浮液的含孢量为 10⁶～10⁷个 /mL时 ,对恶苗病菌的抑制力达 92.33%。通过哈茨木霉菌液和 3种药剂对水稻恶苗病菌抑制效果的比较 ,哈茨木霉孢子悬浮液含孢量为 10⁷个 /mL与施保克质量浓度 1μg/mL的抑菌效果接近 ,分别为 76.7%、75.4%。显微摄影结果显示 ,哈茨木霉以附着胞附着在恶苗病菌菌丝上 ,然后穿透菌丝在其内生长 ,或与恶苗病菌的菌丝平行生长 ,然后再侵入病菌内寄生。     

Study on the Changes of Reproductive Hormone and Regulation of Follicle Development During the Reproductive Cycle of Yili Goose in Xinjiang

This experiment was aimed to study the changes of reproductive hormone and regulation of follicle development during the reproductive cycle of Yili goose in Xinjiang. 2 years old Yili goose were chosen which were in good health and similar weight. 32 samples were randomly selected to take blood samples for hormone detection during the laying, broodiness and ceased periods, and 15 samples were randomly selected to gather ovarian tissue for the observation of the development of ovary and follicular. The results showed that during the laying period, the GnRH of the Yili goose was extremely significantly higher than the broodiness and ceased periods of 16.78% and 58.29% (P<0.01);PRL was significantly lower than the broodiness period of 13.00% (P<0.05), and was extremely significantly higher than the ceased period of 28.94% (P<0.01); FSH was higher than the broodiness period of 6.98% (P>0.05),and was extremely significantly higher than the ceased period of 21.12% (P<0.01); LH was extremely significantly lower than the broodiness period of 8.38% (P<0.01),and was extremely significantly higher than the ceased period of 16.84% (P<0.01); E₂ were extremely significantly higher than that the broodiness and ceased periods of 29.80% and 112.40% (P<0.01); P₄ levels were extremely significantly higher than the broodiness and ceased periods of 28.89% and 30.34% (P<0.01). The ovary volume and weight of Yili goose were extremely significantly higher than that of broodiness and ceased periods (P<0.01). In the laying period, the mature follicles of Yili goose were significantly larger than that of the broodiness and resting periods, the number of secondary follicles was more and the number of primary follicles was lesser. Ovarian growth and mature follicle atresia, granule layer shrinkage, inward depression, cytoplasmic cavity, follicular atrophy at all levels in the broodiness period. The follicle cytoplasmic cavity of the ovary was larger in the ceased period than broodiness period, and inward concave further deeper, a large number of primordial follicles evenly arranged. Thus, during the laying period, the responses of GnRH, E₂ and LH of Yili goose were active and played a leading role on the development of ovarian and follicular. PRL and LH maintained a high level of secretion, cooperated to maintain the broodiness behavior during the broodiness period.     

灰茶尺蠖触角感器的扫描电镜观察

使用扫描电子显微镜对灰茶尺蠖Ectropis grisescens成虫触角感器的种类、形态和分布进行了观察。结果表明:灰茶尺蠖触角上主要存在3种感器,分别为毛形感器(sensilla trichodea, ST)、刺形感器(sensilla chaetica, SC)和栓锥形感器(sensilla styloconica, SS)。感器的种类和分布在雌雄成虫触角上没有差异。