由Phytophthora infestans引起的晚疫病是马铃薯的毁灭性病害。明确现有马铃薯品种或育种材料含有的晚疫病抗病基因，对于抗病育种和合理利用不同抗病基因防治马铃薯晚疫病具有重要意义。根据“基因对基因”学说，马铃薯无毒基因与抗病基因产物互作会产生典型过敏反应（HR）。理论上利用病原菌无毒基因可以鉴定马铃薯是否含有相应抗病基因。本研究尝试利用农杆菌注射技术，在马铃薯叶片中瞬时表达晚疫病菌无毒基因（Avr），根据过敏反应发生情况，进而推断马铃薯品种/育种材料所含相应抗病基因（R）。研究结果显示：适合马铃薯瞬时表达的农杆菌浓度为0.5（OD600），马铃薯材料农杆菌瞬时表达存在明显的基因型和叶龄依赖性，充分展开的幼嫩叶片适合农杆菌瞬时表达。Avr1, Avr2, Avr3a和Avr4在含有相应抗病基因的马铃薯鉴别寄主上瞬时表达能够产生HR，表明无毒基因注射鉴定结果是可靠的。无毒基因注射鉴定结果与抗病基因特异引物PCR扩增结果在不同基因型材料上有时并不一致，反映了这两种方法各有局限性，若能将这两种方法结合使用，则会提高鉴定结果的准确性。

Late blight, caused by Phytophthora infestans, is a devastating disease of potato. It is important to distinguish what kind of R contained in potato cultivar or potato breeding materials for potato late blight resistant breeding and control late blight through management of different R genes. According to the‘gene-for-gene’hypothesis, the interaction between pathogen avirulence genes(Avr)and resistant genes(R)wil lead to hypersensitive response(HR)in potato. So, it is possible to distinguish host R gene by Avr gene expression in potato leaves. In this study, efforts were made to distinguish potato R genes by transient expressing P. infestans Avr gene in potato leaves using agro-infiltration methods. The result revealed that 0.5 optical density at 600 (OD600) was suitable for potato agro-infiltration. Agro-infiltration mediated Avr expression efficiency depended on potato genotype and leaf age. Ful expanded younger leaves were fit for Agro-infiltration mediated transient Avr

【目的】鉴定稻瘟病菌(Magnaporthe oryzae)的无毒基因型,了解无毒基因在不同地区流行菌株中的分布情况,为品种布局提供参考。【方法】根据已经克隆且与稻瘟病菌致病性相关的6个无毒基因序列设计引物,选取辽宁省稻瘟病常发区的26株稻瘟病菌单孢菌株,提取各菌株DNA样本作为模板,进行PCR扩增。通过琼脂糖凝胶电泳及PCR产物测序,对6个无毒基因PCR产物进行碱基和氨基酸序列的分析比较。对琼脂糖凝胶电泳未出条带的,设计不同引物进行验证性试验。【结果】在PCR产物电泳检测中,Avr1-CO39、Avr-pia和Avr-pii没有产物条带,对这3个无毒基因设计验证引物,其PCR产物电泳检测仍没有条带出现,其结果证明辽宁省各水稻主产区流行稻瘟病菌中多不携带Avr1-CO39、Avr-pia和Avr-pii;对于其他3个无毒基因AvrPiz-t、Avr-pik和Avr-pita则有特异性扩增产物,说明这3个无毒基因在各稻区稻瘟病菌中以不同突变类型及不同频率出现。其中,与Pi2、Pi9和Piz-t对应的AvrPiz-t,分别在22个菌株的中被检测到,且有21个菌株的序列与其序列一致,说明该基因遗传相对稳定,也间接证明携带Pi2、Pi9和Piz-t的水稻品种在辽宁地区的广谱抗性;与基因组序列不同的16号菌株,在DNA序列192 bp处发生一个单碱基C的缺失,从而导致移码突变,且碱基突变导致氨基酸序列至72位氨基酸时提前终止,而使该基因编码的蛋白质失去无毒基因的功能。与Pik、Pik-p、Pik-m和pik-s对应的Avr-pik,电泳检测结果表明各菌株均有特异性条带出现,经测序验证等位基因序列分为4种类型(B、D、F、G),其中12个菌株携带可为Pik或其等位基因Pik-m和pik-p所识别的D类型;9个菌株携带B类型,该等位基因曾被报道,但是否具有无毒基因的功能仍未验证;另有2个菌株携带F类型等位基因,该基因为首次发现,并分别出现在丹东和盘锦地区。其特点在于与D类型基因间存在143(A/G)的碱基差异,氨基酸序列翻译结果显示其为错义突变,即48(G/D);而其余3个菌株携带G类型等位基因,该基因亦属首次发现,且仅出现在抚顺新宾地区,碱基序列与D类型存在168(G/A)的差异,导致翻译提前终止,基因功能丧失。对于Avr-pita,26个菌株特异性扩增产物一致,但测序检测到5

[Objective]The objective of this study is to identify avirulence genes (Avr-genes), which existed in the prevalentfungus of Magnaporthe oryzae, understand the distribution of Avr-genes in the epidemic strains of different regions, and to provide references for rice cultivars distribution.[Method]According to the sequences of 6 Avr-genes which had been cloned and were associated with the rice blast pathogenicity, the primers were designed to amplify the target sequences of the epidemic rice blast fungus in Liaoning Province. The DNA, extracted from the 26 strains of rice blast fungi, was used as a template for PCR amplification. By analyzing the PCR products by AGE (agarose gel electrophoresis) and sequencing, the sequences of base and amino acid for each Avr-gene were examined and compared. Some candidate primers were designed to check out the Avr-gene which had no PCR product.[Result]There was no PCR product when amplified Avr1-CO39, Avr-pia and Avr-pii by the primers, and there was a

紫甘薯块根富含花色苷，天然、安全、无毒，具有强大的抗氧化、抗突变和降血糖、保肝和抗高血压等作用。对近年来国内外在紫甘薯花色苷的组成与结构、花色苷生物合成的结构基因和调节基因等方面的研究成果进行了综述，并对下一步的研究重点进行了展望。

There is abundance natural, safe and non-toxic anthocyanin in tuberous roots of purple sweet potato, which providing mighty anti-oxidation and anti-mutation, as well as the effect of hypoglycemic, protecting liver and anti-hypertension. The article summarized the research results in China and abroad about the components and construction of anthocyanin in purple sweet potato, as well as the structural genes and regulatory genes effecting anthocyanin biosynthesis. Then, the key points in further researches were prospected.

为了提高金盏菊植物对环境中重金属（Hg）污染的修复效果，构建 merA 重组表达载体质粒，再经农杆菌介导法建立遗传转化体系筛选出转 merA 基因的金盏菊植株，研究其对汞的富集率。结果表明：转 merA 基因金盏菊对土壤中 Hg2＋的转移率提高84％，而且大部分 Hg2＋在 merA 基因作用下，转化为无毒的 Hg 挥发到空气中，提高了其忍受 HgCl2的浓度；在 Hg2＋胁迫下，转基因植株中脯氨酸含量、SOD 酶活性、POD 酶活性及 MDA 含量均有不同程度的提高。转基因金盏菊具有较强的抗重金属汞胁迫的能力。

The remediation effect of transgenic C.officinalis with merA gene obtained by Agrobacterium-mediated genetic transformation system on mercury pollution was analyzed to study the mercury enrichment rate.The results showed that the 84% Hg2+ in soil can be transported into leaves of transgenic C.officinalis with merA gene and then most Hg2+ in leaves can be converted into nontoxic Hg under the action of merA gene,which can increase its tolerance to HgCl2 stress.The proline and MDA content,and activity of SOD and POD of transgenic C.officinalis with merA gene are increased under Hg2+ stress to varying degrees.In conclusion,transgenic C.officinalis with merA gene has the higher tolerance to Hg2+ stress.

肝癌是我国常见的恶性肿瘤,针对肝癌的传统治疗方法疗效欠佳、患者预后较差.近10余年来,随着分子生物学技术的发展,肝癌的基因治疗成为该领域新的研究方向和热点,其中以腺病毒为载体的单纯疱疹病毒胸苷激酶自杀基因系统（ADV-tk）对肝癌的治疗研究开展得最早也最为广泛.其原理是把单纯疱疹病毒胸苷激酶基因通过腺病毒导入细胞内,利用其产生的酶将无毒的药物前体更昔洛韦（GCV）转变成细胞毒性产物,从而杀死肝癌细胞.多项动物实验和临床研究结果表明：ADV-tk/GCV系统是治疗肝癌的有效方法.本文总结近年来ADV-tk/GCV系统在肝癌治疗中取得的研究进展,分别从载体的发展过程、基因的作用机制、导入方式、增强杀伤效力、降低肝毒性以及相关动物模型的改进等几个方面进行综述.

Liver cancer is one of the most common malignant tumors in China.The efficacy of traditional treatment for liver cancer is unsatisfactory,and the prognosis of the patients is poor.In recent 10 years,with the development of the molecular biological techniques,genetic therapy has become a new and promising approach for liver cancer.Of which,adenovirus mediated herpes simplex virus thymidine kinase (ADV-tk) for the treatment of liver cancer is widely applied.The enzyme secreted by ADV-tk transformed the prodrug gancyclovir (GCV) to the cytotoxic agents and thus to kill the liver cancer cells.The results of multiple animal and clinical experiments showed that ADV-tk/GCV is effective for the treatment of liver cancer.In this article,the recent progress of ADV-tk/GCV in the treatment of liver cancer was reviewed.