中国商陆抗病毒蛋白基因的克隆及其转化辣椒的研究

本研究的目的是获得抗病毒病的辣椒材料,为选育抗病毒病辣椒品种奠定基础。根据美洲商陆抗病毒蛋白基因序列设计引物,从中国商陆（Phytolacca acinosa）叶片基因组DNA扩增克隆缺失无毒型中国商陆抗病毒蛋白基因（PacPAP1,构建该基因的植物表达载体,采用农杆菌介导的叶盘法转化辣椒,对转基因植株进行分子检测、TMV及CMV的人工接种鉴定,接种25d后,统计发病情况。结果表明：克隆得到缺失无毒型PAP基因PacPAP,大小为714 pb,与美洲商陆抗病毒蛋白基因（PamPAP）的同源性为99.7%;得到了PacPAP1整合入辣椒基因组中的阳性植株;接种TMV、CMV阴性对照植株都明显发病,转PacPAP1基因植株未出现症状,说明转PacPAP1辣椒植株可获得抗TMV和CMV材料。     

Outbreak of Philophthalmus gralli in four greater rheas (Rhea americana)

Using slit‐lamp biomicroscopy, conjunctival biopsy, and morphological identification, a flock of four Greater rheas (Rhea americana) in Arizona were diagnosed with conjunctivitis secondary to Philophthalmus gralli (P. gralli) infection. Aquatic snails from the exhibit’s water source were identified as Melanoides tuberculatus, a known vector for P. gralli. Comparison of partial sequences of DNA regions from P. gralli adults removed from the rheas and metacercariae from the aquatic snails demonstrated a 100% match, confirming the source of infection. The flock was divided into two treatment groups: the most severely affected rheas received both manual removal of trematodes and praziquantel 1% ointment OU q12 h and the least severely affected rheas were only given praziquantel 1% ointment OU q12 h. The rheas were permanently relocated away from the infected water source and aquatic snails. Initial resolution was seen at 17 weeks in the most severely affected rhea, which had 675 adult P. gralli removed and topical praziquantel. The two rheas that only received topical praziquantel showed resolution within 3 and 15 weeks. Current recommendations for treating P. gralli include: manual removal of trematodes, topical praziquantel 1% ointment, and relocation away from infected water sources and aquatic snails.     

Effect of pre-incubation humidity and temperature treatment on the in vitro germination of avocado pollen grains

Optimum in vitro germination of pollen grain of the avocado cultivars Fuerte, Nabal, Ettinger, Bacon and Zutano occurred at 25 °C. However, there were significant differences between cultivars in percentage germination and relative humidity (RH) requirements for optimum pollen grain growth. The most sensitive cultivar to relative humidity was Fuerte, in which the germination of pollen grain rose from 11.4%, at 40% RH, to about 50%, after one hour at 100% RH. The germination% of Nabal pollen grain was already high at 40% RH and was not increased by higher relative humidity. Increased relative humidity also helped to sustain the viability of avocado pollen. At 30 °C and 5% RH the pollen grains of Fuerte quickly lost its ability to germinate, at 40% RH for 1 hour, germination was reduced spectacularly compared to pollen kept in saturated with moisture environment where it was not affected the first 24 hours. The effects of temperature and relative humidity on fruit-set and yield of avocado are discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Periplaneta americana peptide decreases apoptosis of pig-ovary granulosa cells induced by H2O2 through FoxO1

Oxidative stress can induce apoptosis of granulosa cells and lead to follicular atresia, thereby reducing the number of pigs giving birth. The aim of this study was to investigate the protective effect of Periplaneta americana peptide (PAP) on the apoptosis of the granulosa cells of pig ovaries (PGCs) induced by hydrogen peroxide (H₂O₂) via FoxO1. PGCs were treated with H₂O₂ to establish a cell apoptosis model. Cell viability was measured using the cell counting kit-8 (CCK-8) assay, and cell apoptosis was detected using flow cytometry. The malondialdehyde (MDA) level and nitric oxide (NO) content were detected to reflect the oxidative stress. Western blotting, qRT-PCR and overexpression were undertaken to determine the expression of FoxO1 and caspase-3, and immunofluorescence was used to detect FoxO1 in the nucleus and cytoplasm. PGCs were treated with 100 μM H₂O₂ for 6 hr, which resulted in oxidative damage and apoptosis and an apoptosis rate for PGCs of 32.95%. Next, PGCs were treated with 400 μg/ml PAP for 24 hr to repair the apoptosis induced by H₂O₂. PAP improved cell viability in H₂O₂-stimulated PGCs, the increased MDA level and NO content caused by H₂O₂ stimulation were reversed and the apoptotic rate of PGCs was reduced. The qRT-PCR and Western blotting results indicated that PAP decreased the H₂O₂-induced apoptosis and the expression of FoxO1 and caspase-3 in PGCs. The effect of PAP was the same following FoxO1 overexpression. FoxO1 was expressed in the nucleus when stimulated by H₂O₂ or overexpression; however, it migrated to the cytoplasm following PAP treatment. PAP decreased the apoptosis of PGCs induced by H₂O₂ by regulating FoxO1 expression and nuclear translocation.     

金边绿玉麻的组织培养

金边绿玉麻高10-15cm的侧芽,在MS 6-BA3．0mg/L(单位下同) IAA1．0培养基中,可诱导产生愈伤和芽丛,其增殖培养,适宜的培养基为MS 6-BA l．0 IAA0．3 Y0．6(多效唑),每30d增殖率达3．0左右．切割下的不定芽在MS IBA0．3培养基中,根的诱导率(生根率)达93％。     

Cloning and Sequencing of the Pokeweed Antiviral Protein Gene and Its Expression in E. coli

The total RNA was isolated from pokeweed (Phytolacca americana ) leaves using the method of guanidine isothiocyanite and used as a template to amplify the deleted mutant pokeweed antiviral protein (PAP) gene by RT-PCR and then the gene was cloned into the pGEMR-T vector. The sequencing results showed that the PAP gene consisted of 711nt, which was 99.6% identical to the PAP gene reported by Lin et al (1991). The IPTG-inducible expression vector containing the PAP gene was constructed and transferred into the E. coli strain BL21 (DE3)-plysS. A specific protein was produced after induction with 0.4m mol/L IPTG and its molecular weight was 26ku. The results of the double diffusion on the agar plate and the western blotting test showed that the protein produced in E. coli was highly identical with the PAP extracted by a Frenchman from French pokeweed leaves. These revealed that PAP gene was actually achieved and exactly expressed in E . coli.     

美国白蜡树绿叶期扦插生根试验

采用美国白蜡树绿叶期1年生带叶的木质化枝条作插穗、1的生根粉作生根促进剂、(石至)石作扦插基质、夜间补光处理的方法进行扦插试验,平均生根率达85～90,当年生苗高达85 cm左右.     

剑麻遗传转化体系的建立与优化

以剑麻无菌苗叶片为试材,研究了潮霉素（Hyg）、菌液浓度、侵染时间、乙酰丁香酮（As）及羧苄青霉素（Car）等因素对剑麻遗传转化的影响。结果表明,剑麻最适遗传转化体系为：Hyg选择浓度为25 mg/L,OD600=0.6左右的农杆菌菌液,侵染时间为10～15 min,As浓度为200μmol/L,Car浓度为400 mg/L。     

金边龙舌兰的开花与栽培繁殖研究(英文)

金边龙舌兰是多年生草本植物。记述了其分类分布、形态特征、生活习性、开花结果、栽培繁殖及日常用途等知识,为金边龙舌兰的开发利用提供了借鉴。     

辛硫磷与溴氰菊酯混配对防治美洲大蠊增效机理研究

利用电生理和生化方法,以美洲大蠊为材料,研究辛硫磷与溴氰菊酯混用的增效机理。结果表明:辛硫磷与溴氰菊酯混配,特别是1×10-5mol/L辛硫磷与1×10-6mol/L溴氰菊酯混配,兴奋性突触后电位(EPSP)开始兴奋时间比单用溴氰菊酯缩短了67%,阻断时间缩短了36 min以上,说明混剂提高了乙酰胆碱的释放和在突触间隙的积累。生化分析认为,混剂增效的原因之一是提高了对靶标酶——AchE的抑制。