甜瓜果实特异性表达黄瓜素基因启动子区的克隆和序列分析

黄瓜素(cucum is in)是甜瓜类枯草杆菌丝氨酸蛋白酶,其表达具有果实特异性.应用PCR方法从甜瓜基因组DNA中扩增出该基因自转录起始位点至上游一段长310 bp片段,并将其克隆到pUC 19载体中,序列分析表明该序列与已报道的相应序列完全相同,且具有TATA-box、CAAT-box、G-box、I-box-like和增强子元件TGTCACA等功能域,具有典型的果实特异性启动子特征.成功获得甜瓜果实特异性表达基因的启动子,为下一步实现外源基因在甜瓜果实中特异表达奠定基础.

甜瓜黄瓜素基因启动子表达载体的构建及分析

该研究构建了由黄瓜素基因5′端310bp启动子序列驱动β-葡萄糖醛酸酶(GUS)报告基因的植物表达载体pPZP-CGN,通过花粉管通道法将植物表达载体pPZP-CGN导入甜瓜,并采用荧光法定量测定转基因植株中GUS活性。结果显示,gus基因在果实中高表达,而在根、茎、叶等组织中表达活性很低,表明黄瓜素基因上游310bp启动子具有指导外源基因在果实中高效特异表达的特性。

甜瓜黄瓜素基因启动子的序列及元件分析

本研究对甜瓜黄瓜素基因启动子进行了元件分析。讨论以采集的甜瓜叶片为试验材料,采用CTAB法提取植物DNA,并利用PCR扩增技术,成功获得了黄瓜素启动子的基因序列,利用DNAman、DNAstar等软件进行处理和分析序列结果,并采用Plant CARE和PLACE对启动子的元件进行生物信息学元件分析。分析结果表明:黄瓜素启动子与AY055805、LN713264、LN681897的同源性为100%、99%、99%。其序列含有多种高等植物果实启动子通用的顺式作用元件TATA-Box、CAAT-Box和光响应元件,同时部分序列还也参与了ABA、VP1反应和水杨酸反应。甜瓜黄瓜素启动子的研究为下一步利用该启动子进行深入的果实基因工程研究提供材料基础和理论依据。

Effects of Reducing Mn^(2+),Cu^(2+) and Zn^(2+) Concentrations in Nutrient Solution on SOD Activity in Grafted Cucumber Seedling Leaves Under Cold Stress

[Objective] The research aimed to study the effects of reducing the concentrations of Mn2＋, Cu2＋ or Zn2＋ in nutrient solution on the contents of Mn, Cu and Zn of grafted cucumber leaves as well as on the SOD and isozymes activities and the rate of electrolytic leakage under cold stress. [Method] Nutrient solution containing low concentrations of Mn2＋, Cu2＋ or Zn2＋ were used for cultivating the grafted cucumber seedlings for the cold treatment. [Result] The results indicated that reducing the concentrations of Mn2＋, Cu2＋ or Zn2＋ in nutrient solution had declined the SOD activities in grafted cucumber leaves and increased the rates of electrolytic leakage. Mn2＋ had shown the maximum influences, followed by Cu2＋, and Zn2＋ had shown the minimum influences. [Conclusion] Cold resistance of cucumber can be improved by changing the contents of Mn2＋, Cu2＋ or Zn2＋ in nutrient solution.

Integrated Control of Tetranychus cinnabarinus by Five Kinds of Pesticides Combined with Neoseiulus cucumeris

To find the method for the integrated control of Tetranychus cinnabarinus （carmine spider mite） and delay its resistance to pesticides, the joint actions of each of five commonly pesticides （abamectin, azadirachtin, matrine, pyrethrins and imidacloprid）, and Neoseiulus cucumeris （Oudemans）, the natural enemy of cotton red spider mite, were studied. N. cucumeris was released after application of pesticides for seven days （for 0,3% azadirachtin EC） or six days （for other pesticides）. The results showed that the combined action of 1.8% abamectin EW （1：8 000） and N. cucumeris had the best control efficacy of 96.63% at 20 days after N. cucumeris releasing. The control efficacies of N. cucumeds and 0.3% azadirachtin EC （1：250） were 59.7% and 90.1% after one day and 20 days, respectively, after N. cucumeris releasing. The control efficacy of N. cucumeris and 0.5% matrine AS （1：2 000） was 82.65% at 20 days after N. cucumeris releasing. The results provide options for sustainable control of T. cinnabarinus and for the delay of pesticides resistance.