双重PCR检测马铃薯晚疫病和环腐病方法的建立

通过克隆马铃薯环腐病菌和晚疫病菌转录间隔区(ITS)序列,并对测序结果进行同源性比较,选取差异位点分别设计了两对引物P.IN1/P.IN2和C.IN1/C.IN2,并检测了引物的特异性及方法的灵敏度。引物P.IN1/P.IN2可扩增出1条363bp马铃薯晚疫病菌的特异性条带,在DNA水平上其灵敏度达18fg/μL;引物C.IN1/C.IN2可扩增出1条218bp马铃薯环腐病菌的特异性条带,在细菌数上检测灵敏度为104 cfu/mL。混合这两对引物构建双重PCR反应体系,能从马铃薯环腐病菌和晚疫病菌的混合DNA及感染这两种菌的马铃薯植株中同时扩增到363bp和218bp的特异片段。实现了同时对马铃薯晚疫病菌和环腐病菌的快速可靠检测。     

吡虫啉及其复配剂不同施药方法对油菜蚜虫控制效果

2006-2009年,采用拌种和播种沟施药的方法,于河南郑州研究了新烟碱类杀虫剂吡虫啉及吡虫啉.辛硫磷复配剂对油菜蚜虫的全程控制效果。小区试验结果表明,用吡虫啉有效成分5g/kg种子拌种仅能有效控制苗期油菜蚜虫的危害,防治效果为90.79%～100%,对中后期油菜蚜虫控制效果不明显;用吡虫啉600g/hm2及吡虫啉.辛硫磷复配剂1320g/hm2采用播种沟施药对油菜蚜虫的防治效果分别为91.82%～94.49%和90.68%～95.34%;试验示范验证防治效果在92%以上,持效期长达7个月以上,可以达到一次用药控制油菜整个生长期蚜虫为害的效果,减少了用药次数,此方法简单易行、保护环境、保护天敌,是一种简化高效的具有推广价值的油菜蚜虫可持续控制技术。     

Potential gene flow from transgenic rice (Oryza sativa L.) to different weedy rice (Oryza sativa f. spontanea) accessions based on reproductive compatibility

BACKGROUND: The possibility of gene flow from transgenic crops to wild relatives may be affected by reproductive capacity between them. The potential gene flow from two transgenic rice lines containing the bar gene to five accessions of weedy rice (WR1–WR5) was determined through examination of reproductive compatibility under controlled pollination. RESULTS: The pollen grain germination of two transgenic rice lines on the stigma of all weedy rice, rice pollen tube growth down the style and entry into the weedy rice ovary were similar to self‐pollination in weedy rice. However, delayed double fertilisation and embryo abortion in crosses between WR2 and Y0003 were observed. Seed sets between transgenic rice lines and weedy rice varied from 8 to 76%. Although repeated pollination increased seed set significantly, the rank of the seed set between the weedy rice accessions and rice lines was not changed. The germination rates of F₁ hybrids were similar or greater compared with respective females. All F₁ plants expressed glufosinate resistance in the presence of glufosinate selection pressure. CONCLUSIONS: The frequency of gene flow between different weedy rice accessions and transgenic herbicide‐resistant rice may differ owing to different reproductive compatibility. This result suggests that, when wild relatives are selected as experimental materials for assessing the gene flow of transgenic rice, it is necessary to address the compatibility between transgenic rice and wild relatives. Copyright © 2009 Society of Chemical Industry     

苹果小吉丁虫在陕西省潜在危害性风险分析和管理对策

苹果小吉丁虫是果树危险性蛀干害虫之一,已给国内部分地区的栽植苹果以及新疆的野苹果造成了巨大的损失。目前陕西部分地区已发现苹果小吉丁虫危害,为明确苹果小吉丁虫在陕西省的主要苹果产区的风险程度,以制定科学合理的管理对策。参考国际上有害生物风险分析程序和分析方法,结合苹果小吉丁虫寄主在陕西省的分布情况,对苹果小吉丁虫在陕西省的风险性进行了综合评估。苹果小吉丁虫在陕西省的风险性R值为2.42,该吉丁虫在陕西省属于高度危险性有害生物。针对苹果小吉丁虫在陕西省的高度危险性,提出了加强检疫、监测预防和风险管理等的管理对策。     

Molecular cloning and expression of kidney‐type glutaminase from common carp (Cyprinus carpio) and its up‐regulation by glutamine in primary culture enterocyte

Glutaminase (GLS) is the key enzyme of glutamine (Gln) metabolism and utilization. In this study, a cDNA encoding GLS protein was identified from common carp Cyprinus carpio intestine. The open reading frame of GLS cDNA encodes a polypeptide of 595 amino acids, which shows a high similarity with its zebrafish Danio rerio counterpart. Bioinformatic analysis showed the protein belongs to kidney‐type GLS. The putative protein has glutaminase domain and ankyrin repeats domain, which are highly conserved among vertebrate orthologues. Real‐time quantitative PCR analysis revealed that the abundance of GLS mRNA was the highest in the white muscle, followed by the brain, eyeball and pituitary. Glutaminase was ubiquitously expressed in all intestinal segments of common carp. The activity of GLS did not distribute uniformly along the entire length of the intestine. In primary culture enterocyte, and the expression of GLS mRNA is up‐regulated quickly and effectively by Gln.     

草莓快速繁殖技术的研究

本文探讨了利用茎尖离体培养法进行草莓快速繁殖的方法。主要摸索了激素对试管苗的分化、生根的影响及移栽成活的技术要点。     

表达鸡传染性法氏囊病病毒VP2重组鸡痘病毒的构建

含ＩＢＤＶ保护性抗原ＶＰ２的质粒,以ＳｐｈＩ消化,然后用Ｔ４噬菌体ＤＮＡ聚合酶将末端补平,产物纯化后再以ＳａｌＩ消化一次,经再次纯化后与ＳｍａＩ和ＳａｌＩ分步酶切的鸡痘病毒插入载体ｐＦＧ１１７５－１相连,使唤基因顺向插入到载体Ｐ７５启动子下游,得到含ＩＢＤ ＶＰ２基因的重组插入载体ｐＦＧ ＶＰ２。     

长吻鮠精子保存液在人工授精中的应用试验

分别在安徽长江流域的无为和淮河流域的滁州进行长吻鮠的人工繁殖,使用LRH-A2、PG、混合激素B型、DOM 进行催产,从长江捕获的亲鱼立即催产,其催产率为66．67％,受精率80％-85％,经培育选择后催产率为83．33％,受精率65%- 87％。在滁州经多年培育的亲鱼催产率为100％,受精率78％-95％。精子保存液在精子保存和人工授精中使用效果良好。在冰桶内保存,20-30 h仍能保持良好的受精能力。     

磷化氢熏蒸对桔小实蝇氧化代谢体系的影响研究

为阐明磷化氢的生化毒理机制,本试验以重要检疫性害虫桔小实蝇(Bactrocera dorsalis Hendel.)为试虫,研究了25℃下磷化氢熏蒸对桔小实蝇3龄幼虫的毒力以及磷化氢熏蒸不同时间对桔小实蝇3龄幼虫氧化代谢体系的影响。结果显示：（1）25℃条件下磷化氢熏蒸桔小实蝇3龄幼虫48 h的LD50、LD90、LD99分别为0.28、0.37、0.46 mg/L;（2）磷化氢熏蒸抑制了桔小实蝇的呼吸作用;（3）随熏蒸时间延长,桔小实蝇体内过氧化氢含量和脂质过氧化水平不断上升,氧化压力逐渐增加;（4）磷化氢抑制了过氧化氢酶活性,最大抑制率为55.99%,增强了过氧化物酶活性、超氧化物歧化酶活性及抑制羟自由基能力,最大提高率分别为141.48%、130.21%、155.90%。本研究确认了磷化氢熏蒸杀灭桔小实蝇的有效性,其作用机制可能是通过抑制过氧化氢酶活性进而加剧活性氧积累实现的。