High frequency embryogenic callus induction and plant regeneration from mature caryposis of big bluestem and little bluestem

Big bluestem (Andropogon gerardii Vitman) and little bluestem [Schizachyrium scoparium (Michaux) Nash.] are native to the North America and are important forage grasses and ornamental grasses. Both grasses are proposed as ideal biomass producers for cellulosic ethanol production. To apply genetic transformation, which is an important tool for incorporating desirable agronomic traits into plants to both species, however requires an efficient and reproducible regeneration protocol. We used mature caryopses from big and little bluestem as explants and tested the effect of various combinations of 2, 4-dichlorophenoxyacetic acid (2, 4-D) (1, 2, 3, 4 or 5 mg l⁻¹) and kinetin (KT) (0, 0.1 or 0.2 mg l⁻¹) on embryogenic callus induction with LS as the basal medium. The highest percentage of embryogenic calli induction occurred on medium containing 2, 4-D alone at 2 mg l⁻¹ for ‘Bison’ and on medium containing 4 mg l⁻¹ 2, 4-D alone for ‘Bonilla’ big bluestem. For little bluestem, the highest percentage of embryogenic callus induction occurred on medium containing 3 mg l⁻¹ 2, 4-D plus 0.1 mg l⁻¹ kinetin, suggesting that addition of KT is beneficial. Shoot regeneration took place on LS basal medium without any plant growth regulator for both species, although the addition of KT increased both regeneration frequency and the number of shoots produced per callus. Rooting of shoots reaching about 2 cm long occurred readily with or without α-naphthaleneacetic acid (NAA). Rooted plantlets were all successfully established in the soil.     

对虾白斑综合征病毒研究概况

对虾白斑综合征病毒(WSSV)引起世界范围内养殖对虾的急性、致死性传染病,造成对虾养殖业的巨大损失,在我国这种情况尤为严重。WSSV是同一类紧密相关的无包埋体类杆状病毒,与杆状病毒并无同源性。2001年,中国和荷兰科学家分别报道了WSSV的全基因组DNA序列,启动了WSSV后基因组研究。已有多种检测方法用于WSSV的检测和致病机理的研究。WSSV感染宿主谱广,以多种传播途径感染宿主,目前还没有用于病毒增殖的细胞系,但已有成熟的动物模型,从而方便了对WSSV的研究。本文就该病毒的命名、病毒特性、检测方法、病毒感染增殖、感染宿主、传播途径及感染动物模型等方面的研究现状和进展作一综述。     

分泌抗鸡Ig单抗杂交瘤细胞系的建立与单抗生物学特性的鉴定

从传染性法氏囊病病毒（ＩＢＤＶ）单克隆抗体（ＭｃＡｂ）杂交瘤细胞系１Ｄ１０、１Ｇ１中得到了３株能稳定分泌抗鸡免疫球蛋白（Ｉｇ）ＭｃＡｂ的杂交瘤细胞系１Ｂ７、１Ｄ７、３Ｇ６。其抗体类和亚类均属ＩｇＧ２ｂ，腹水的ＥＬＩＳＡ效价≥１∶２５６００，琼扩效价为１∶８～１∶１６。３株ＭｃＡｂ能与鸡血清中的Ｉｇ出现沉淀反应，琼扩在２４ｈ以内出现清晰的沉淀线，而不能和鸭、鸽、鹌鹑等禽类及异种动物血清出现沉淀线。纯化的鸡ＩｇＧ、ＩｇＭ经ＳＤＳ－ＰＡＧＥ后，分别用纯化并经辣根过氧化物酶（ＨＰＲ）标记的１Ｂ７、１Ｄ７、３Ｇ６单抗酶结合物进行免疫印迹试验证明，３株单抗识别的均为ＩｇＧ、ＩｇＭ分子的轻链     

蛋鸡血清硷性磷酸酶变化规律的研究

本试验以京白Ⅲ系为试验材料,测定了四个时期的血清AL P活性,研究了其变化规律。结果表明,AL P同工酶型快型酶活性显著高于慢型。年龄、同工酶型及年龄×同工酶型互作对AL P活性有显著影响。AL P活性与产蛋量间遗传相关表现为,在生长期为正,而产蛋期为负。四个年龄中,AL P做为间接选择指标,以6周龄为最佳.     

流行性出血热病毒气溶胶感染乳鼠的免疫病理学研究

采用免疫组化ＢＡ法，对流行性出血热病毒（ＥＨＦＶ）气溶胶感染乳鼠脏器组织中的病毒动态分布进行了观察，并对其抗体消长进行了测定。结果表明：①ＢＡ法为病理诊断ＥＨＦＶ感染提供了一种准确的手段；②ＥＨＦＶ可通过气溶胶途径感染乳小鼠，乳小鼠感染ＥＨＦＶ的ＬＤ５０值可作为环境浓度的参考值；③ＥＨＦＶ气溶胶途径感染乳鼠后，４ｈ即可在心、肺，１ｄ在嗅球部检出，并可在其组织细胞中增殖，经血循、嗅神经、单核巨噬细胞系统等多途径扩散，造成多组织脏器的泛发性感染。感染后１２～１４ｄ，病毒检出率和病毒量均达峰值。动物自身免疫系统对ＥＨＦＶ有一定的清除能力；④感染乳鼠脑、肺、肾等脏器组织的病变与人类感染ＥＨＦＶ的病变有一定的相似性，提示可作为人ＥＨＦＶ的模型动物；⑤抗原检出率高于抗体检出率，ＥＬＩＳＡ法检出率高于ＩＦＡ法检出率。     

马传贫驴白细胞弱毒疫苗株跨膜蛋白主要免疫决定区基因的克隆与表达

从感染驴白细胞的马传贫驴白细胞弱毒疫苗株前病毒DNA中克隆了编码跨膜蛋白主要免疫决定区(TMIR)的基因，并在大肠杆菌中进行了表达。所表达的融合蛋白有一部分是可溶的，其氨基端带有6个组氨酸的标签，因此可以用固定化金属离子亲和层析法在非变性条件下进行纯化。在间接酶联免疫吸附试验(ELISA)和免疫印迹试验中，重组的TMIR蛋白可与马传贫阳性血清样品发生反应，而与健康马血清无任何反应。这表明该重组蛋白具有良好的抗原性和特异性，可用于马传贫弱毒疫苗株在体内外复制、接种马体内免疫应答及马传贫诊断的研究。     

鸡空肠弯曲菌灭活苗的制备及其免疫保护试验研究

用分离到的空肠弯曲菌制备鸡空肠弯曲菌甲醛灭活疫苗 ,并进行了该病的免疫保护实验。制备 2× 10 9个 / m L浓度的细菌悬液 ,4m L / L浓度的甲醛溶液作用 12 h灭活后制成空肠弯曲菌灭活苗 ,免疫 2周龄的 SPF雏鸡 ,14 d后 ,进行攻击实验 ,观察其临床症状 ;并分别于感染后 7d和 14 d捕杀 ,观察病理学变化及进行细菌分离实验以揭示其保护率。结果表明 ,鸡空肠弯曲菌灭活苗的保护率可达 10 0 % ,说明该苗经口免疫后安全可靠 ,无毒副作用     

鸡白介素18基因原核表达质粒构建

根据已发表的江西土鸡白介素 -1 8(Ch IL-1 8) c DNA编码基因序列设计引物 ,用PCR技术从 p MDCh IL-1 8质粒扩增出编码鸡IL-1 8成熟蛋白基因 ,重组于 p BV2 2 0表达载体上 ,将重组质粒转化大肠杆菌 JM1 0 9(DE3 ) ,转化子经温度诱导的表达产物 ,SDS-PAGE电泳鉴定约 2万 ,N端开头 1 5个氨基酸序列测定分析 ,证明获得了鸡 IL-1 8成熟蛋白 ,为今后深入研究鸡 IL -1 8的生物学特性及其临床应用打下了基础     

Spermatophore cryopreservation and artificial insemination of black tiger shrimp, Penaeus monodon (Fabricius)

To develop an appropriate cryopreservation protocol for spermatophores of black tiger shrimp, Penaeus monodon, three cryoprotectants (dimethyl sulphoxide (DMSO), methanol (MeOH) and ethylene glycol (EG)) at two concentrations (5% and 10%) were examined. Artificial implantation of spermatophores was also carried out to assess the fertilizing ability of fresh and post‐thaw spermatophores. Spermatophores were collected during consecutive regenerations (15‐day intervals) and assessed for qualitative and quantitative changes and also for fertilizing ability by implantation. The mean fertilization rate for artificial insemination using post‐thaw spermatophore was 79.9±3.7%, lower than the fertilization rates observed for artificial implantation using fresh spermatophore and natural mating. Mean hatch rates for fresh spermatophore, frozen‐thawed spermatophore and natural mating were 88.8±0.6%, 87.8±0.4% and 88.3±0.5%, respectively; and there was no difference among the three groups. The mean fertilization rate of spermatophores collected during the first stripping was higher (90.6±0.6) than during the second stripping (85.7±2.6), but the mean hatch rate was not different between the two strippings. The highest mean sperm viability (79.7±0.4%) was obtained from DMSO (5%), with no survival observed in the 10% MeOH treatment. Spermatophore weight, total sperm count and percentage of abnormal sperm were not different between spermatophores collected at the first and second stripping. This is the first study to report high fertilization and hatch rates from cryopreserved spermatophore using artificial implantation of spermatophore before spawning.     

新型的人工饲料防腐剂ADA

将新型防腐剂——ＡＤＡ添入家蚕人工饲料中，对在高温多湿养蚕环境下，易引起人工饲料霉变的酵母菌群、格兰氏阴性菌群和霉菌群有明显的抑制作用，可延长人工饲料的存放和使用时间，而不影响其适口性、颜色、香味与成份，防腐效果明显优于苯甲酸、山梨酸、脱氢乙酸、丙酸等．实验证明：人工饲料中添加０．２％的ＡＤＡ能使人工饲料在高温多湿的养蚕环境下保持一个月不发霉变质，并对蚕体的生长发育无不良影响