SSR技术及其在果树上的应用

SSR(Simple sequence repeat)技术以其丰富的多态性、共显性遗传、重复性好和操作简便等优点日益受到重视,已成为植物遗传和育种研究中不可缺少的分子标记。对SSR技术的原理和特点作了简要的介绍,较详细地分析了如何获得SSR引物,特别是综述了果树上SSR引物的研究现状,同时将其与其它几种主要的分子标记进行了比较分析,认为SSR标记检测的位点多态性水平明显高于RFLP,而且重复性优于RAPD;着重介绍了SSR技术在果树种质资源和构建果树遗传图谱及基因定位等研究中的应用现状;指出SSR技术将在果树科研上起到重要的作用。     

Effect of heat shock precondition on reperfusion arrhythmia in rats

AIM:To investigate the effect of the heat shock response on the reperfusion arrhythmias(RAs) and the possible mechanism involved. METHODS:Fifty-five Sprague Dawley rats were randomly divided into 2 groups: the heat shock group (group H,n=29) and the control group (group C,n=26). The rats in group H were preconditioned with heat shock 24 hours before, and that in group C were not. The hearts of 16 rats in group H and 16 in group C were exercised and mounted on a non-circulating Langendorff perfusion apparatus and perfused retrogradely with modified K-H buffer and mimic ischemia/reperfusion was applied. Additionally, conventional intracellular microelectrode techniques were used for recording such electrophysiological parameters as resting potential(RP), action potential amplitude(APA), over shot(OS), maximum depolarization velocity(Vmax) of the hearts of other 13 rats in group H and 10 in group C. RESULTS:①Prior heat stress significantly decreased reperfusion arrhythmia. ②The amount of CK release in the effluent in group H was much less than that in group C. ③Myocardial HSP70 content was elevated significantly in group H. ④Heat stress significantly increased myocardial anti-oxydases activity and decreased lipid peroxydative products. Additionally, heat stress significantly reduced the Vmax of action potential. It indicated that rapid Na⁺ channel of papillary muscles may be inhibited by heat shock. The degree of change of Vmax after ischemia in H group was significantly less than that in group C. And the time of reperfusoin with Tyrode's solution till the action potential appeared as large as that before perfusion with mimic ischemic solution is shorter in group H than in group C. CONCLUSION:Heat shock pretreatment markedly reduces ischemia/reperfusion-induced injury of heart and ventricular arrhythmias in rats and this effect may be associated with the inhibition of rapid Na⁺ channel of papillary muscles by heat shock and the increase in myocardial HSP70 and anti-oxydase activity.     

Effects of Taoren Honghua Jian on the hyperlipidemia without symptom

AIM: To examine the effect of traditional chinese medicine recipe, Taoren Honghua(semen persicae-flos carthami) decoction, on hyperlipidemia without symptom. METHODS: The plasma TC, TG, LDL, HDL, apolipoprotein(Apo) A, Apo Bof the patients with hyperlipidemia without symptom were measured using automatic analyzer (shimadiu CL-7200), the production of nitric oxide(NO) was detected by Greiss reaction, and SODactivity and MDAformation were examined using o-trihydroxy benzene and barbituric methotheds, respectively. RESULTS: After oral administration of Taoren Honghua decoction, the plasma levels of TC, TG, LDL, and MDAof the patients were markedly decreased, however, the plasma levels of ApoA, HDL, SODand NOwere significantly increased and almost no change was detected in the plasma level of Apo B. In control group, it was found that although the plasma level of TC, TG and LDL were decreased ( P<0.05 ) and ApoAas well as HDLwere increased, ApoB, SOD, MDA and NO production were all unchanged. CONCLUSION: The traditional chinese medicine recipe, Taoren Honghua decoction,has a significant therapeutic effect on patients with hyperlipidemia by removing blood stasis, promoting qi circulation and in turn reducing blood lipid level.     

大果型鲜食枣新品种‘大白铃’

 ‘大白铃’为大果型鲜食枣优良新品种。果实特大, 平均单果质量24. 5 ~ 25. 9 g, 最大80 g,整齐, 外形美观, 品质优良。早实, 丰产, 抗逆性强。     

外源绿原酸对苹果抗病相关酶的影响

采用离体培养方法对富士、红星和八棱海棠进行绿原酸处理,研究苹果在外源绿原酸处理下POD、PPO、PAL活性的动态变化以及绿原酸含量的变化。结果表明,富士以0.0001%绿原酸、红星以0.0008%绿原酸、八棱海棠以0.0004%绿原酸处理后,其PAL活性均显著提高,苹果体内也因此而提高了绿原酸水平。     

对鳞翅目害虫有活性的cry1C基因的克隆和表达

在鉴定苏云金芽孢杆菌(Bacillus thuringiensis，简称Bt)Btc001菌株cry基因型的基础上，构建了Btc001菌株质粒DNA HindⅢ片段的文库，并利用聚合酶链式反应-限制性酶切片段多态性(PCIR-RFIP)方法筛选出含有crylC6全长基因的13．5kb大片段，酶切分析得到该片段的物理图谱，BamHI和EcoRI切完成了6．5kb含全长基因的亚克隆，并对这条6．5kb片段亚克隆、测序，序列在国际核酸序列数据库(GenBank)登记(AY007686)，并由Bt杀虫晶体蛋白基因国际命名委员会命名为cry1Cb2基因。根据序列设计了一对用于扩增全长基因的引物S581CB和S381CB，扩增产物插入表达载体pET-21b中，诱导后在大肠杆菌BL21(DE3)中获得高效表达。表达产物对小菜蛾(Plutella xylostella)表现出较高活性，LC50达到7．9μg/ml。     

梨火疫细菌实时荧光PCR和诱捕PCR-ELISA检测方法的建立

根据梨火疫细菌中独特而保守存在的质粒pEA29，设计了1对引物和3条探针，建立了实时荧光PCR检测方法和诱捕PCR-ELISA检测方法。实时荧光PCR采用带荧光标记的核酸杂交探针，边扩增边检测，步骤简单，不需PCR后处理，可避免假阳性和交叉污染；诱捕PCR-ELISA检测方法只需简单处理的样品就能检测，减少了核酸不纯出现的漏检，由于增加了核酸杂交探针，可不需凝胶电泳EB染色检测，不会出现假阳性问题。     

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。     

寄主植物对桃蚜羧酸酯酶和乙酰胆碱酯酶的诱导作用

在１９９５～１９９６年研究了寄主植物对桃蚜［Ｍｙｚｕｓｐｅｒｓｉｃａｅ（Ｓｕｌｚｅｒ）］羧酸酯酶（ＣａｒＥ）和乙酰胆碱酯酶（ＡＣｈＥ）的诱导作用。在试验的甘蓝、茄子和桃树３种寄主植物中，取食甘蓝的桃蚜种群ＣａｒＥ和ＡＣｈＥ活性最高，取食茄子和桃树的桃蚜种群ＣａｒＥ活性没有明显不同，而ＡＣｈＥ活性取食茄子的桃蚜种群明显高于取食桃树的种群。ＣａｒＥ与底物的亲和力是桃树＞茄子＞甘蓝，而ＡＣｈＥ与底物的亲和力则是甘蓝＞茄子＞桃树。ＡＣｈＥ与毒扁豆碱的双分子速率常数（Ｋｉ）值大小顺序为甘蓝＞桃树＞茄子     

菜豆萎蔫病菌的种子检验鉴定技术研究

本研究对我国一类检疫性有害生物菜豆萎蔫病菌Ｃｕｒｔｏｂａｃｔｅｒｉｕｍｆｌａｃｃｕｍｆａ－ｃｉｅｎｓｐｖ．ｆｌａｃｕｍｆａｃｉｅｎｓ（Ｃｆ）进行种子检测鉴定技术的研究，发现５２３，ＮＢＹ＋ＴＴＣ和ＮＢＹ＋刚果红三种培养基均可作为分离培养基；对带菌种子分离的灵敏度为１．８×１０２ｃｆｕ／ｍｌ，病叶柄和茎是分离的最佳部位，病菌可通过种子传给种苗引起种苗发病，发病时间为７～２０天，针刺接种是Ｃｆ最有效的接种方法，接种发病率为１００％。使用标准化的Ｂｉｏｌｏｇ鉴定系统鉴定病原，在种水平的鉴定准确率达９８．５％。