Influences of chronic hypoxia on the gene expression of Kv1.3,Kv2.1 and Kv3.1 induced by acute hypoxia in PASMC of rats

AIM and METHODS: Total RNA was extracted from 6th rat subcultured pulmonary artery smooth muscle cells(PASMC) exposed to continual chronic hypoxia or normoxia and the effects of chronic hypoxia on the changes of Kv1.3,Kv2.1,Kv3.1 mRNA in cultured PASMC induced by acute hypoxia were studied by semiquantitative RT-PCR in vitro. RESULTS:①Kv1.3,Kv2.1,Kv3.1 genes were found to be expressed in PASMC of rats exposed either to hypoxia or normxia.②The expression of Kv2.1 and Kv3.1 in 6th subcultured of PASMC in normaxia group could be upregulated by exposure to acute hypoxia,the levels of Kv2.1 and Kv3.1 mRNA were significantly increased from 0.646±0.092, 0.782±0.104 to 1.059±0.134, 0.985±0.116,respectively (P<0.01,n=5). ③PASMC cultured continuously in chronic hypoxia for 6 subcultures and then exposed to normoxia for 12 h,thereafter the expression of Kv2.1 and Kv3.1 were downregulated by acute hypoxia for 6 hours.The level of Kv2.1 mRNA was significantly decreased from 1.008±0.117 to 0.649±0.097 (P<0.01,n=5). CONCLUSION:Kv2.1,Kv3.1 genes might be oxygen sensitive genes.Chronic hypoxia might change the response of these Kv genes of PASMC to acute hypoxia and down-regulate its expression,which might probably decrease the role of Kv in HPV.     

High efficient expression of human endostatin in E.coli and its antiangiogensis activity

AIM: To examine the expression of human endostatin in E.coli, produce its fusion protein antibody and observe its biological activity. METHODS: Endostatin gene was amplified by polymerase chain reaction,recombined with plasmid vector pGEX-2T and induced expression with IPTG.The protein activity was tested by endothelial cell proliferation inhibitory assay.Inclusion body crudely purified was used to generate polyclonal antibody to detect its expression at mouse's liver and kidney etc. RESULTS: The protein expressed was 20kD after digestion by thrombin,it appeared the anti-angiogenesis activity and Western blotting indicated the expression of endostatin in liver and kidney of mouse. CONCLUSION: The successful expression of human endostatin and the preparation of polycolonal antibody indicated its potential application in anti-angiogenesis therapy and diagnosis tumors.     

达县分葱田甜菜夜蛾发生规律初步研究

采用系统调查方法研究了达县地区分葱田甜菜夜蛾的生活史、发生与环境的关系。甜菜夜蛾在达县分葱田1a发生6代,以三至五代为害最重,世代重叠。其数量消长与虫源、夏秋季气候、天敌、栽培管理等因素密切相关。通过调查分析提出本县初始虫源非本地虫源。     

肥城桃组培苗诱导、基因转化及其增殖

红里、白里是肥城桃的两个优良品种,但不耐贮藏。为试图利用基因工程解决这一问题,先通过组培将其胚、胚乳、子叶的愈伤组织分别诱导再生植株和芽剥离出茎尖培育成苗。再将肥城桃反义PG基因,通过农杆菌介导转化组培苗,转化苗在含卡那霉素培养基上筛选,获得抗卡那霉素的转基因苗。用研制的增殖培养基培养4～7周,一个苗可生长出14～18分枝的苗,解决了转基因苗获得率低的问题,也满足了对转基因株系进行检测和从组培室到温棚到田间果园过渡栽培和芽接的需求,提高了繁育速率。     

磷化氢熏蒸处理对嗜卷书虱不同虫态的致死作用

实验室条件下系统研究了磷化氢(PH3)对储物害虫嗜卷书虱Liposcelis bostrychophila卵、各龄若虫和成虫的致死作用,并选用PH3间歇熏蒸以及PH3与气调交替处理等措施对嗜卷书虱进行处理,比较了不同处理措施对嗜卷书虱种群的控制效果。结果表明,PH3熏蒸处理对嗜卷书虱各虫态有不同的致死效果。对卵而言,24、72和120 h熏蒸处理的LC50分别为0.137、0.045和0.035 mg/L;而24 h熏蒸处理对若虫的LC50在4.285~7.364 μg/L之间,对成虫的LC50为20.404 μg/L;采用25 μg/L的PH3进行24 h熏蒸处理,间隔10 d后再分别进行第2次和第3次熏蒸处理,可以完全控制嗜卷书虱的发生。采用PH3 (12 μg/L)和气调(体积比例为35% CO2,1% O2,64% N2)交替处理能够延缓嗜卷书虱种群抗性的发展,交替处理3~5次可以完全控制嗜卷书虱的发生。     

黑杨萎蔫枝叶对棉铃虫生殖行为的影响

研究了黑杨(Populus nigra)萎蔫枝叶及其气味对棉铃虫成虫的隐藏部位、产卵部位、交配率及产卵量等的影响，并对黑杨萎蔫枝把在田间诱集到的棉铃虫雌蛾进行了解剖观察。结果表明，在田间，黑杨萎蔫枝把诱集到的棉铃虫93．3％已经交配。在气候箱中，放置黑杨萎蔫枝叶的环境中配对饲养的棉铃虫，经过1个暗期和2个暗期后，其交配率与对照环境中的均无显著差异。在光期开始时，有51．5％的成虫隐藏在饲养笼内所设置的黑杨叶子圆片下，对照纱布圆片下仅有16．7％。叶子圆片所在区域的日平均落卵量显著大于对照区。黑杨萎蔫枝叶及其气味对棉铃虫的产卵前期、产卵持续期、成虫寿命和产下50％的卵所用的时间无显著影响，但饲养笼内有黑杨枝把或黑杨叶片气味存在时，棉铃虫的产卵量比模拟枝把和对照显著提高。     

制约稻粒黑粉病菌冬孢子萌发的关键因素

在相同条件下，从稻田直接收集的病粒中的稻粒黑粉病菌厚垣孢子不能萌发，而取之于仓库中的孢子能萌发。为探明其制约因素，对该病菌进行了光照和浸水时间试验。研究结果表明：稻粒黑粉病菌冬孢子形成后，必须经光照射后，方可进入休眠期；度过5～6个月休眠期的冬孢子，必须浸水48h以上才开始复苏；复苏后的冬孢子必须在光照条件下才能萌发。未完成后熟作用的冬孢子和完成后熟但尚未复苏的冬孢子即使在光照条件下也不能萌发。即稻粒黑粉病菌冬孢子必须经历后熟、休眠、复苏三个阶段后，在适宜的光照与温湿条件下，方可萌发。光照对其冬孢子具有双重作用，促使后熟进入休眠和刺激萌发。     

温湿度调控对番茄灰霉病菌产生的细胞壁降解酶的影响

 番茄灰霉病菌在致病过程中能够产生4种细胞壁降解酶,以PMG酶活性最高,其次是β-葡萄糖苷酶和PG酶,Cx最少。灰霉病菌在不同温度下侵染番茄叶片时产生的致病酶活性不同,4种酶在20℃时表现了最高的活性,15℃次之,当温度达到25℃时,各种酶的活性都急剧下降,随着温度的再升高,酶活更低。随着湿度的增高,病菌产生的细胞壁降解酶的活性也增加,当相对湿度达到90%以上时,4种酶的活性也达到最高。温湿度对番茄灰霉病菌产生细胞壁降解酶的影响趋势,与其对发病的影响趋势是一致的。     

利用RT-PCR检测库尔勒香梨苹果茎痘病毒的研究

 以库尔勒香梨新鲜、冷藏、冷冻叶片和皮层为材料,对提取双链RNA (dsRNA)的2种方法和提取总RNA的3种方法进行了分析比较,并对总RNA的提取方法进行了改进,获得了纯度较高、完整性较好的dsRNA和总RNA,在此基础上进行了反转录(RT)和PCR扩增。在国内首次完成了对苹果茎痘病毒(ASPV)的RT-PCR检测,建立了ASPV有效RT-PCR反应体系。用此体系扩增到ASPV一个长约316 bp的片段。实验表明以dsRNA和总RNA为模板均能成功进行RT-PCR检测,且dsRNA优于总RNA。     

粉红女士苹果品质特性分析评价

观察测定了粉红女士苹果成熟采收时和常温贮藏后果实的外观、抗性、果实硬度、总酸、可溶性固形物、维生素C、蒸腾失水率、出汁率、果汁密度以及果汁性状;全面分析评价了该品种的外在、内在、贮藏品质和果汁加工特性。结果表明,粉红女士苹果是一个综合性状优良、耐贮藏的鲜食、果汁加工兼用型苹果新品种。