【目的】建立一种快速、灵敏、特异的检测甘蔗条点病毒(sugarcane striate virus,SStrV)的SYBR Green I荧光定量PCR方法。【方法】从SStrV基因组保守区域序列设计特异性扩增引物,构建含有SStrV基因组序列的重组质粒pMD19-T-SStrV-qN作...【目的】建立一种快速、灵敏、特异的检测甘蔗条点病毒(sugarcane striate virus,SStrV)的SYBR Green I荧光定量PCR方法。【方法】从SStrV基因组保守区域序列设计特异性扩增引物,构建含有SStrV基因组序列的重组质粒pMD19-T-SStrV-qN作为阳性质粒标准品,以其为模板建立SStrV荧光定量PCR检测方法,并对该方法的灵敏性、特异性、稳定性进行了测试,随后用该方法对甘蔗不同组织部位中SStrV载量进行检测。【结果】将含有SStrV基因组序列的重组质粒按10倍比稀释成标准品,将其作为模板进行荧光定量PCR,获得标准曲线y=-3.337 x+38.197,相关系数r2=0.999,说明Cq值与标准品浓度拷贝数的对数呈线性关系;建立的荧光定量PCR最低可以检测到13拷贝重组质粒/μL,是普通PCR灵敏度的100倍。该方法能特异的检测SStrV,特异性高,组内和组间的变异系数在0.13%-0.94%之间,表明该方法重复性良好。SStrV的载量在甘蔗的不同组织部位中差异较大,+4叶中SStrV的载量最高,与其他组织部位达到了显著差异。【结论】建立了能灵敏特异检测SStrV的SYBR Green I荧光定量PCR方法,明确了+4叶是甘蔗中SStrV检测的最佳采样部位。展开更多
[Objective] The paper was to explore the flowering habits and fertility of Gaozhou common wild rice （Oryza rufipogon Griff.） in Lingshui of Hainan Province. [Method] The flowering habits, pollen fertility and seed s...[Objective] The paper was to explore the flowering habits and fertility of Gaozhou common wild rice （Oryza rufipogon Griff.） in Lingshui of Hainan Province. [Method] The flowering habits, pollen fertility and seed setting rate of Gaozhou common wild rice in Lingshui of Hainan Province were studied, and compared with the flowering habits of common wild rice in Hainan Province. [Result] Gaozhou common wild rice was able to flower twice a year. The first flowering stage was from mid-April to mid-June for about 60 d, and the daily peak flowering dates were from May 13 to May 22. The daily flowering time was mainly between 11：20 and 13：00, and the peak flowering was at 11：30. The second flowering stage was from late August to late October for about 45 d, and the peak flowering dates lasted from September 8 to September 18. The daily flowering time was mainly from 11：15 to 12：50, and the daily peak flowering was at 11：30. In both flowering stages, the panicles had the same flowering characteristics. The stigma exsertion reached a high value of 95.37% with single （46.23%） and dual （49.14%） stigmas exsertion respectively. A spikelet remained open for 1.5-2.5 h, the angle of spikelet opening varied from 29.9° to 32.4°. At the two flowering stage the average pollen fertility rates were 89.67% and 85.62%, the average seed setting rates were 58.81% and 55.79%, respectively. Hainan population flowered only once （from mid-October to mid-December）, this had significant difference with the performance of Gaozhou common wild rice which flowered twice a year. [Conclusion] The results provided basis for the research on gene flow frequency from transgenic rice to wild rice and its relationship with flowering habits of common wild rice.展开更多
文摘【目的】建立一种快速、灵敏、特异的检测甘蔗条点病毒(sugarcane striate virus,SStrV)的SYBR Green I荧光定量PCR方法。【方法】从SStrV基因组保守区域序列设计特异性扩增引物,构建含有SStrV基因组序列的重组质粒pMD19-T-SStrV-qN作为阳性质粒标准品,以其为模板建立SStrV荧光定量PCR检测方法,并对该方法的灵敏性、特异性、稳定性进行了测试,随后用该方法对甘蔗不同组织部位中SStrV载量进行检测。【结果】将含有SStrV基因组序列的重组质粒按10倍比稀释成标准品,将其作为模板进行荧光定量PCR,获得标准曲线y=-3.337 x+38.197,相关系数r2=0.999,说明Cq值与标准品浓度拷贝数的对数呈线性关系;建立的荧光定量PCR最低可以检测到13拷贝重组质粒/μL,是普通PCR灵敏度的100倍。该方法能特异的检测SStrV,特异性高,组内和组间的变异系数在0.13%-0.94%之间,表明该方法重复性良好。SStrV的载量在甘蔗的不同组织部位中差异较大,+4叶中SStrV的载量最高,与其他组织部位达到了显著差异。【结论】建立了能灵敏特异检测SStrV的SYBR Green I荧光定量PCR方法,明确了+4叶是甘蔗中SStrV检测的最佳采样部位。
基金Supported by Special Project of GM(2009ZX08011-010B,2008XZ08011-011)~~
文摘[Objective] The paper was to explore the flowering habits and fertility of Gaozhou common wild rice （Oryza rufipogon Griff.） in Lingshui of Hainan Province. [Method] The flowering habits, pollen fertility and seed setting rate of Gaozhou common wild rice in Lingshui of Hainan Province were studied, and compared with the flowering habits of common wild rice in Hainan Province. [Result] Gaozhou common wild rice was able to flower twice a year. The first flowering stage was from mid-April to mid-June for about 60 d, and the daily peak flowering dates were from May 13 to May 22. The daily flowering time was mainly between 11：20 and 13：00, and the peak flowering was at 11：30. The second flowering stage was from late August to late October for about 45 d, and the peak flowering dates lasted from September 8 to September 18. The daily flowering time was mainly from 11：15 to 12：50, and the daily peak flowering was at 11：30. In both flowering stages, the panicles had the same flowering characteristics. The stigma exsertion reached a high value of 95.37% with single （46.23%） and dual （49.14%） stigmas exsertion respectively. A spikelet remained open for 1.5-2.5 h, the angle of spikelet opening varied from 29.9° to 32.4°. At the two flowering stage the average pollen fertility rates were 89.67% and 85.62%, the average seed setting rates were 58.81% and 55.79%, respectively. Hainan population flowered only once （from mid-October to mid-December）, this had significant difference with the performance of Gaozhou common wild rice which flowered twice a year. [Conclusion] The results provided basis for the research on gene flow frequency from transgenic rice to wild rice and its relationship with flowering habits of common wild rice.
